Unit 1 INTRODUCTION TO FISH MUSCLE FUNCTION 1

Chapter 1: Introduction and historical developments in low

temperature preservation of fish (1)
1.1.1.1. Historical developments in low temperature preservation of
foods
1.1.1.2. Demand for low temperature preserved fish
Chapter 2: Structure and function of fish muscle (1)
1.2.1.1. Lipids
2.1.2. Proteins (2)
1.2.1.3. N – containing extractives
1.2.1.4. Vitamins and Minerals
Chapter 3: Postmortem changes in fish (3)
1.3.1 Changes in raw fresh fish
1.3.2 Changes in eating quality
1.3.3 Autolytic changes (4)
1.3.3.2 Autolysis and nucleotide catabolism (4)
Chapter 4: Bacteriological changes (6)
1.4.1 Native bacterial flora of fishes
1.4.2 Factors that influence the growth of microorganisms (7)
Chapter 5: Spoilage of fish (7)
1.5.1 Spoilage of fish
1.3.3.2 Autolysis and nucleotide catabolism

Unit 2 Chilling of fish 8

Chapter 1: Fresh fish handling (8)
2.1.1. Introduction to Icing
2.1.2. Advantages of chilling of fish with ice (9)
2.1.3. Chilled storage
2.1.4 Storage method (10)
Chapter 2: Calcultation of the ice requirement for cooling fish (11)
2.2.1.Calculation of the ice requirement of cooling fish
2.2.1.1. Heat Requirements (12)
2.2.1.2. Calculation of the ice requirement for the storage of fish (13)
Chapter 3: Manufacturing of different types of Ice (13)
2.3.1.1. Block Ice (14)
2.3.1.2. Flake Ice
2.3.1.3. Plate Ice (15)
2.3.1.4.Tube Ice
Chapter 4: Super chilling (0ºC to - 4ºC) (16)
2.4.1.1. Refrigerated Sea Water (RSW)
2.4.1.2. Chilled sea water (CSW) (17)
2.4.1.3. Slurry Ice in Fish Preservation
2.4.1.4. Cooling and Storage Tests
Chapter 5: Application of ozone/chlorine in seafood processing (18)
2.5.1. Application of ozone in seafood processing
2.5.2. Use of Chlorine in Fish Processing
2.5.2.1.Concentration of chlorine used in Fish processing (19)
2.5.2.2. Marginal Chlorination
2.5.2.3. Super Chlorination (19)
2.5.2.3.1. Break-point chlorination
2.5.2.3.2. In-plant Chlorination (20)
Unit 3 Freezing preservation of fish 20
Chapter 1: Principle of freezing (20)
3.1.1. Specific heat
3.1.2. Thermal conductivity
3.1.4 Latent heat (21)
3.1.5 Principles of freezing
Chapter 2: Physical, chemical and thermodynamic properties
of refrigerants (23)
3.2.1. Refrigerant
3.2.2. Thermodynamic properties
3.2.3. Latent heat of vaporization (24)
3.2.4 Physical properties of refrigerants
3.2.5. Refrigerant (25)
3.2.6 Freon group (26)
3.2.7 Secondary refrigerant (27)
3.2.8. Physical characteristics of refrigerants
3.2.9. Chemical properties of refrigerants (28)
3.2.10. Chemical properties (29)
3.2.11. Economic and thermodynamic properties
Chapter 3: Types of freezer (29)
3.3.1.1. Air blast freezers (30)
3.3.1.2. Plate freezers
3.3.1.3. Liquid nitrogen freezer (31)
3.3.1.4. Carbon dioxide freezer
3.3.1.5. Immersion freezers (32)
Chapter 4: Methods of protective treatments (32)
3.4.1. Brining
3.4.2. Advantages of brine treatment
3.4.3. Use of Polyphosphate in freezing of fish (33)
3.4.4. Methods of using antioxidants (34)
Chapter 5: Calculation of freezer refrigeration load (35)
3.5.1. Calculation of freezer refrigeration load
3.5.1.1. Freezing time (36)
3.5.1.2. Calculation of freezing times
3.5.1.3. Coding, Packing and Storage (37)
Chapter 6: Freeze drying fish (38)
3.6.2. Theory of freeze drying (39)
3.6.3. Principle of the equipment
3.6.4. Freeze drying process (40)

Unit 4 Thawing of fish 40

Chapter 1: Methods of fish thawing (40)
4.1.1.1 Thawing in air
4.1.1.2 Water thawing (41)
4.1.1.3 Thawing between heated plates
4.1.1.4 Choice of method (43)

Unit 5 Quality changes during frozen storage 43

Chapter 1: Quality changes in fish during freezing & frozen storage-
physical changes (43)
5.1.1 Quality changes in fish during freezing
5.1.1.1 Ice formation
5.1.1.2 Desiccation (44)
5.1.1.3 Discolouration
5.1.1.4 Fat oxidation during freezing (45)
5.1.1.5 Factors limiting storage life (46)
5.1.1.6 Freeze Denaturation-I (47)
 5.1.1.7 Freeze Denaturation-II (47)
Chapter 2: Microbial growth at low temperature (48)
5.2.1 Microbial growth at low temperature

Unit 6 Cold stores and containers 49

Chapter 1: Construction of cold store (49)
6.1 Construction of cold store
6.1.1 Shape and size (50)
6.1.2 Insulation
6.1.3 Air ingress (51)
6.1.4 General layout
Chapter 2: Refrigerated containers (52)
6.2.1 Refrigerated Containers (52)
6.2.2 Loading
Chapter 3: Good handling and shipping practices (53)
6.3.1 Fresh fish
6.3.2 Frozen Fish

Unit 7 Packaging methods 53

Chapter 1: Packaging for fresh fish handling (53)
7.1.1 Packaging for Fresh Fish Handling
7.1.1.1 Moulded polystyrene Fish Box (54)
7.1.1.2 Refrigerants and Coolants
7.1.1.3 Guidelines for air shipment of sea food (55)
Chapter 2: Packaging of frozen fishery products (55)
7.2.1.1 Primary packages (56)
7.2.1.2 Flexible films
7.2.1.3 Shipping container (57)
7.2.1.4 Shrink packaging
7.2.1.5 Requirements for packaging materials
Chapter 3: Vaccum packaging and modified atmosphere packaging
and storage (58)
7.3.1 Vaccum packaging
7.3.2 Modified atmospheric packaging and storage (59)

Unit 8 HACCP 59
Chapter 1: Hazard analysis and critical control point- A detailed
approach (59)
8.1.1 Introduction to the codex Alimentarius commission
8.1.2 Work of the codex alimentarius commission
8.1.3 History of HACCP (60)
8.1.4 Principles of the HACCP system
8.1.5 The HACCP team (62)
8.1.6 Form. 2 Product ingredients and incoming material
8.1.7 Hazard analysis: (Principle 1) (63)
8.1.8 How to conduct a hazard analysis (65)
8.1.9 Hazard assessment
8.1.10 Critical control points: Determination (Principle 2) (66)
8.1.11 Establish critical limits for each CCP (Principle 3)
8.1.12 Establish a monitoring system for each CCP (Principle 4) (67)
8.1.13 Establish corrective actions (Principle 5)
8.1.14 Establish verification procedures (Principle 6) (68)
8.1.15 Establish Documentation and Recorded keeping (Principle 7)
Chapter 2: Physical, chemical and biological hazards in seafood (68)
8.2.2 Hazard Analysis
8.2.2.1 Health Hazards from Pests (69)
8.2.2.2 Prevention (70)
8.2.3 Chemical hazards-analysis (70)
8.2.4 Biological hazards-analysis (71)
Chapter 3: Important biotoxin in seafood (71)
8.3.1 Tetradotoxin
8.3.2 Ciguatera toxin (72)
8.3.3 Paralytic shellfish poinsoning
8.3.4 Diarrhetic shellfish poinsoning(DSP) (73)
8.3.5 Neurotoxic shellfish poisoning(NSP)
8.3.6 Amnesic shellfish poisoning(ASP)
8.3.7 Control of disease caused by biotoxin
8.3.8 Toxic metals (74)
8.3.9 Antibiotic Residues
Chapter 4: Histamine (75)
8.4.1 Biogenic Amines
8.4.2 Control of Disease caused by biogenic amines (76)
Preservation of Fish by Chilling and Freezing Technology

Unit 1: Introduction to fish muscle function

Chapter 1: Introduction and historical developments in low temperature preservation of fish

1.1.1.Introduction
Several food preservation techniques were evolved in order to enable man to use food produced at various places
from age old times. Such methods of preservation of food are sun drying, salting, keeping food in ice, smoking etc.
Fish is a highly perishable commodity as it contains good amount of nutrients and hence there is a greater need to
preserve it. Without preservation it is impossible to transport and distribute fish to interior places in good condition.
1.1.1.1. Historical developments in low temperature preservation of foods
In olden days, in places which were covered with snow or ice, the flesh of hunted animals and fish were covered with
ice thereby fish was frozen and quality is preserved. However, freezing of fish/ foods by artificial methods is a recent
development.
Towards the end of 19th century compressors were developed and because of this machine, novel methods of fish
freezing were developed. First patent on freezing of fish using cold air was given to V.Benzamine of England in 1842.
In the year 1861, similar patent was awarded to E. Piper of U.S.A. Who used salt ice mixture to freeze food. Russians
were pioneers in freezing fish by using machines. They used compressors on board the fishing vessel to freeze fish
and also built a freezing plant (factory) in a place called Astrakhan. In 1898 immersion freezing system was
developed for freezing fish. Nikhoidahl, a scientist from Norway developed spray freezing method. In 1925, a
technician named Cook developed contact plate freezer which was further improved by Karensbirdseye in 1929.
Similarly methods of freezing and packing it in consumer packs were developed subsequently.

1.1.1.2. Demand for low temperature preserved fish

In most of the processing methods such as salting, drying, canning etc., changes in the sensory characteristics take
place. Most people like fish with characteristics as existing in freshly caught fish. Therefore, the principal problem
encountered by fishing industry is to preserve fish in such a way that it does not lose its original sensory
characteristics. Even for processing by other methods, initial preservation of fish is done by low temperature
preservation technique such as icing. Hence low temperature preservation of fish has become increasingly popular
in recent years.

Chapter 2: Structure and function of fish muscle

1.2.1. Introduction
Most of the fish muscle tissue is white but, depending on the species, many fish will have a certain amount of
dark tissue of a brown or reddish colour located just under the skin along the side of the body. In pelagic fish, i.e.,
species such as herring and mackerel which swim more or less continuously, up to 48% of the body weight may
consist of dark muscle (Love, 1970). In demersal fish, i.e., species which feed on the bottom and only move
periodically, the amount of dark muscle is very small.From technology point of view, the high lipid content of dark
muscle is important because of problems with rancidity.
The energy source for ATP generation in the light muscle is glycogen, whereas the dark muscle may use lipids. A
major difference is, that the dark muscle contains much more mitochondria than light muscle, thus enabling the dark
muscle to operate an extensive aerobic energy metabolism resulting in CO2 and H2O as the end products. The light
muscle, mostly generating energy by the anaerobic metabolism, accumulates as lactic acid which has to be
transported to the liver for further metabolization. In addition, the dark muscle is reported to possess functions
similar to those are found in the liver.
1.2.1.1. Lipids
The lipids present in teleost fish species can be divided into two major groups: the phospholipids and the
triglycerides. The phospholipids make up the integral structure of the unit membranes in the cells; thus, they are
often called structural lipids. The triglycerides are lipids used for storage of energy in fat depots, usually within
special fat cells surrounded by a phospholipid membrane and a weak collagen network. The triglycerides are often
termed depot fat. A few fish have wax esters as part of their depot fats.
1
 The mobilization of energy is much faster in light muscle than in dark muscle, but the formation of lactic acid
creates fatigue, leaving the muscle unable to work for long periods at maximum speed. Thus, the dark muscle is used
for continuous swimming activities and the light muscle for quick bursts, such as when the fish is about to catch a
prey or to escape a predator.
In elasmobranchs, such as sharks, a significant quantity of the lipid is stored in the liver and may consist of fats
like diacyl-alkyl-glyceryl esters or squalene. Some sharks may have liver oils with a minimum of 80% of the lipid as
unsaponifiable substance, mostly in the form of squalene.
Fish lipids differ from mammalian lipids. The main difference is that fish lipids include up to 40% of long-chain
fatty acids (14-22 carbon atoms) which are highly unsaturated. Mammalian fat will rarely contain more than two
double bonds per fatty acid molecule while the depot fats of fish contain several fatty acids with five or six double
bonds.
2.1.2. Proteins
The proteins in fish muscle tissue can be divided into the following three groups:

1. Structural proteins (actin, myosin, tropomyosin and actomyosin), which constitute 70-80% of the total
protein content (compared with 40% in mammals). These proteins are soluble in neutral salt solutions of
fairly high ionic strength (> 0.5M).
2. Sarcoplasmic proteins (myoalbumin, globulin and enzymes) which are soluble in neutral salt solutions of low
ionic strength (<0.15M). This fraction constitutes 25-30% of the protein.
3. Connective tissue proteins (collagen), which constitute approximately 3% of the protein in teleost and about
10% in elasmobranch (compared with 7% in mammals).

The majority of the sarcoplasmic proteins are enzymes participate in the cell metabolism, such as the anaerobic
energy conversion from glycogen to ATP. If the organelles with in the muscle cells are broken, this protein fraction
may also contain the metabolic enzymes localized inside the endoplasmic reticulum, mitochondria and lysosomes.
The fact that the composition of the sarcoplasmic protein fraction changes when the organelles are broken was
suggested as a method for differentiating fresh from frozen fish, under the assumption that the organelles were
intact until freezing. The proteins in the sarcoplasmic fraction are excellently suited to distinguish between
different fish species, as all the different species have their characteristic band pattern, when separated by the
isoelectric focusing method. Fish protein is an excellent source of all essential amino-acids. In diets based mainly on
cereals, a supplement of fish can raise the biological value significantly.

1.2.1.3. N – containing extractives

The N-containing extractives can be defined as the water-soluble, low molecular weight, nitrogen-
containing compounds of non-protein nature. This NPN-fraction (non-protein nitrogen) constitutes from 9 to 18%
of the total nitrogen in teleosts. The major components in this fraction are: volatile bases such as ammonia and
trimethylamine oxide (TMAO), creatine, free amino-acids, nucleotides and purine bases, and urea in the case of
cartilaginous fishes.

Fish Crustaceans Poultry Mammalian muscle

Compound in mg/100g Cod Herring Shark species Lobster Leg muscle
wet weight
1) Total extractives 1200 1200 3000 5500 1200 3500
2) Total free amino – acids 75 300 100 3000 440 350
Arginine <10 <10 <10 750 <20 <10
Glycine 20 20 20 100-1000 <20 <10
Glutamic acid <10 <10 <10 270 55 36
Histidine <1.0 86 <1.0 -- <10 <10
Proline <1.0 <1.0 <1.0 750 <10 <10
3) Creatine 400 400 300 0 -- 550
4) Betaine 0 0 150 100 -- --
5)Trimethylamine oxide 350 250 500-1000 100 0 0

2
 The amount of TMAO in the muscle tissue depends on the species,
Some Mineral constituents of fish muscle
season, fishing ground, etc. In general, the highest amount is found in
elasmobranchs and squid (75-250 mg N/100g) followed by cod (60- Elements Content (mg/100 g)
120mg N/100g),while flatfish and pelagic fish have the least. Pelagic Sodium 30-134
fish (sardines, tuna, and mackerel) have their highest concentration of
TMAO in the dark muscle while demersal, white fleshed fish have a Potassium 19-502
much higher content in the white muscle. Quantitatively, the main
Calcium 19-881
component of the NPN-fraction is creatine. In resting fish, most of the
creatine is phosphorylated and supplies energy for muscular
Magnesium 4.5-452
contraction.

1.2.1.4. Vitamins and Minerals Phosphorus 68-550

Iron 1-5.6
In general, fish meat is a good source of vitamin B and in the case of
fatty species A and D vitamins. Some freshwater species such as carp
Chlorine 3-761
have high thiaminase activity so the thiamine content in these species
is usually low. Fish meat is regarded as a valuable source of calcium
Iodine 0-2.73
and phosphorus in particular iron, copper and selenium in general.
Saltwater fish have a high content of iodine.

Chapter 3: Postmortem changes in fish

1.3.1 Changes in raw fresh fish

Immediately after death the muscle is totally relaxed and the elastic texture usually persists for some hours,
thereafter the muscle will contract. When it becomes hard and stiffs the whole body becomes inflexible and the fish
is in rigor mortis.The rate in onset and resolution of rigor varies from species to species and is affected by
temperature, handling, size and physical condition of the fish. Rigor mortis starts immediately or shortly after death,
if the fish is starved and the glycogen reserves are depleted, or if the fish is stressed during fishing.

In rigor the fish body will be completely stiff; the filleting yield will be very poor, and rough handling can
cause gaping. If the fillets are made from pre-rigor muscle, it contract freely and the fillets will shorten following the
onset of rigor. Dark muscle may shrink up to 52% and white muscle up to 15% of the original length (Buttkus, 1963).
If the pre-rigor fish is cooked the texture will be very soft and pasty. In contrast, the texture is tough but not dry
when the fish is cooked in rigor. Post-rigor the flesh will become firm, succulent, elastic and best suited for
processing.
1.3.2 Changes in eating quality
Changes in the eating quality of chilled fish during
storage can be assessed by daily organoleptic examination
of the cooked flesh.
Phase 1: The fish is very fresh and has a sweet, sea
weedy and delicate taste. The taste can be very slightly
metallic. In cod, haddock, whiting and flounder, the sweet
taste is maximized 2-3 days after catching.
Phase 2: There is a loss of the characteristic odour
and taste. The flesh becomes neutral but has no off-
flavours. The texture is still pleasant
Phase 3: There is sign of spoilage and a range of
volatile, unpleasant-smelling substances are produced
depending on the fish species and type of spoilage (aerobic,
anaerobic). One of the volatile compounds may be
trimethylamine (TMA) derived from the bacterial reduction
3
of trimethylaminoxide (TMAO). TMA has a very characteristic “fishy” smell. At the beginning of the phase the off-
flavour may be slightly sour, fruity and slightly bitter, especially in fatty fish. During the latter stages sickly sweet,
cabbage-like, ammoniacal, sulphurous and rancid smells develop. The texture becomes either soft or watery of
tough and dry.
Phase 4: The fish can be characterized as spoiled and putrified.

1.3.3 Autolytic changes

Autolysis means “self-digestion”. It has been
known for many years that there are at least two
types of fish spoilage: enzymatic and bacterial.

Muscle enzymes and their activity: At the point of

death, the supply of oxygen to the muscle tissue is
interrupted because the blood is no longer pumped
by the heart and is not circulated through the
gills. Since no oxygen is available for normal
respiration, the production of energy from ingested
nutrients is greatly restricted.
Figure illustrates the normal pathway for the
production of muscle energy in most living teleost
fish (bony finfish). Glycogen (stored carbohydrate) or
fat is oxidized or “burned” by the tissue enzymes in a
series of reactions which ultimately produce carbon
dioxide (CO2), water and the energy-rich organic compound adenosine triphosphate (ATP). This type of respiration
takes place in two stages: an anaerobic and an aerobic stage. The latter depends on the continued presence of
oxygen (O2) which is only available from the circulatory system. Most crustaceans are capable of respiring outside
the aquatic environment by absorption of atmospheric
oxygen for limited period.

Aerobic and anaerobic breakdown of glycogen in

vertebrate & invertebrate muscle tissue.

Under anaerobic conditions, ATP may be synthesized by

two other important pathways from creatine phosphate or
from arginine phosphate. The former source of energy is
restricted to vertebrate muscle (teleost fish) while the
latter is characteristic of some invertebrates such as the
cephalopods (squid and octopus).

In either case, ATP production ceases when the creatine or

arginine phosphates are depleted. It is interesting to note
that octopine is the end-product from the anaerobic
metabolism of cephalopods and is not acidic (unlike
lactate), thus any changes in post mortem pH in such animals are not related to the production of lactic acid from
glycogen. For most teleost fish, glycolysis is the only possible pathway for the production of energy once the heart
stops beating.

After death, when the regeneration ceases, the ATP is rapidly degraded i.e. after death, the anaerobic muscle cannot
maintain its normal level of ATP, and when the intracellular ATP level declines from 7 - 10 µmoles/g to <1.0 µmoles/g
tissue, the muscle enters rigor mortis. Post mortem glycolysis results in the accumulation of lactic acid which in turn
lowers the pH of the muscle. In cod, the pH drops from 6.8 to an ultimate pH of 6.1- 6.5. In some species of fish, the
final pH may be lower: in large mackerel, the ultimate rigor pH may be as low as 5.8 - 6.0 and as low as 5.4 - 5.6 in
tuna and halibut, however such low pH levels are unusual in marine teleosts.
4
 In general, fish muscle contains a relatively low level of glycogen compared to mammals, thus far less lactic
acid is generated after death. Also, the nutritional status of the fish and the amount of stress and exercise encountered
before death will have a significant effect on the levels of stored glycogen and consequently on the ultimate post
mortem pH. As a rule, well-rested, well-fed fish contain more glycogen than exhausted fish. It is show that bleeding of
fish significantly reduced the post mortem production of lactic acid.
The post mortem reduction in pH of fish muscle has an effect on the physical properties of the muscle. As
the pH drops, the net surface charge on the muscle proteins is reduced, causing them to partially denature and lose
some of their water-holding capacity. Muscle tissue in the state of rigor mortis loses its moisture when cooked and
is particularly unsuitable for further processing which involves heating, since heat denaturation enhances the water
loss. Loss of water has a detrimental effect on the texture of fish muscle and it has been shown by Love (1975) that
there is an inverse relationship between muscle toughness and pH, unacceptable levels of toughness (and water-
loss on cooking) occurs at lower pH levels.

1.3.3.2 Autolysis and nucleotide catabolism

Autolysis and nucleotide catabolism
According to the present understanding, rigor mortis is caused by bonding of the myosin heads, extending
radially from the thick microfibrils, to the active centers in actin units of the thin filaments. This leads to the formation
of a rigid structure of interconnected myofilaments. This reaction between these major muscle proteins is made
possible by changes in the regulatory proteins. These changes are in turn induced by an increase in the concentration
of Ca2+ in the sarcoplasm. In a resting muscle, the concentration of free Ca2+ in the sarcoplasm is lower than 10-7 M.
This level is maintained because of the action of various calcium pumps, located in the cell membrane and in the
sarcoplasmic reticulum, which drives the Ca2+ out of the sarcoplasm, against the concentration gradient. The calcium
pumps work at the expense of energy from ATP hydrolysis. After the death of the animal, a gradual depletion of ATP
in the muscles takes place, due to exhaustion of the creatine phosphate and glycogen reserves. This impairs the action
of the calcium pumps and increases the concentration of Ca2+ in the sarcoplasm. At ATP concentrations below 10-4 M
and that of Ca2+ above 10-6 M, rigor mortis sets in. As the individual muscle fibers contain different quantities of ATP,
they do not enter rigor at the same time. Thus, stiffness in a
muscle sets in gradually.
The resolution of rigor is a process still not
completely understood but always results in the subsequent
softening (relaxation) of the muscle tissue and is thought to
be related to the activation of one or more of the naturally-
occurring muscle enzymes, digesting away certain
components of the rigor mortis complex. The softening of
the muscle during resolution of rigor (and eventually
spoilage processes) is coincidental with the autolytic
changes. Among the changes, one of the first to be
recognized was the degradation of ATP-related compounds
in a more-or-less predictable manner after death.

Degradation of ATP to hypoxanthine (Hx) and ribose

Freshness is expressed by K-value. This K-value expresses the relationship between inosine and
hypoxanthine and the total amount of ATP-related compounds:
HxR + Hx
K(%) = ------------------------------------------------------------------------- x 100
ATP + ADP + AMP + IMP + HxR + Hx

Very fresh fish, have low K-values. K-values increase gradually which is species dependent. Inosine is said to
be more or less flavourless, while hypoxanthine has been reported to impart a bitter flavour in spoiling fish (Spinelli,
1965). Hx is considered to have a direct effect on the perceived bitter off-flavour of spoiled fish (Hughes and Jones,
1966). It is now widely accepted that IMP is responsible for the desirable fresh fish flavour which is only present in
top quality seafood.

5
Chapter 4: Bacteriological changes
1.4.1 Native bacterial flora of fishes
The flesh and body fluids of live healthy fish are generally free from bacteria, i.e. sterile. But, even when the fish is
alive, they harbour bacteria, mainly on three sites of their body (i.e.) the
Salinity range of water bodies
slime on the skin surface the gill tissue and the intestine. Bacteria, which are
naturally present on fishes, are called the native bacterial flora of fish. The Water Salinity range
population and nature of such flora depend on the waters from where the Sea water 30-36 ppt
fish were caught, i.e. whether seawater, brackish water or fresh water. Brackish water 10-28 ppt
Seawater has more dissolved salts, i.e. a higher salinity than both brackish Fresh water 0.2-1.0 ppt
and fresh water.
Depending on the salinity and pollution range of waters, the bacterial flora of fishes varies from water to water.
Generally the bacterial population of the fishes from tropical water is in the following ranges .

Bacterial population of brackish water fishes

Bacterial population of marine fishes from tropical waters
Sample Pearl spot Milkfish
Sample Oil sardine Indian mackerel (Etroplus
(Sardinella (Rastrelliger suratensis ) (Chanos chanos)
longiceps) kanagurta) 3 4
Skin with 10 -10 103-105
Skin with slime/cm2 103-107 104-106 slime/cm2
Gills/g 105-108 104-109 Gills/g 104-108 105-108
5 9 5 8
Intestine with 10 -10 10 -10 Intestine with 105-108 106-108
contents/g contents/g

Generally, the bacterial populations on the skin surface

are the least, while bacterial counts in the intestine are Bacterial population of fresh water fishes
the highest. Bacterial counts in the gill tissue are more Sample Rohu (Labeo Mrigal (Cirrhinus
or less between the bacterial counts of skin surface and rohita) mrigala)
the intestine. Bacterial populations exhibit seasonal Skin with 4
10 -10 5
104-105
variations. During warmer months, the counts used to slime/cm2
be higher while during colder seasons, the bacterial
Gills/g 104-106 105-107
counts are lower. Qualitatively, the composition of
bacterial flora of the fishes from the three waters differ Intestine with 105-107 104-106
considerably. Majority of the bacterial flora of marine contents/g
fishes are gram negative, non-spore forming (asporogenous) rods or cocci. Bacterial flora of fresh water fishes is
predominantly gram positive in nature. The microflora of the brackish water fishes are evenly composed of both
gram +ves and gram –ves.

From these examples, it can be seen that in the case of marine fishes, not only there is a larger proportion of gram-
negative bacteria, but also, there is a large number of bacterial types. But for fresh water fishes, the number of
bacterial genera is quite limited.

a. Microbial flora on the skin surface of marine fish:

Gram negative: Vibrio , Pseudomonas , Moraxella , Acinetobacter , Flavobacteria , Aeromonas , Photobacteria.

Gram positive: Arthrobacter , Micrococcus , Bacillus.

b. Bacterial flora on the skin surface of brackish water fish:

Gram negative: Pseudomonas , Alcaligenes , Flavobacter , Moraxella , Vibrio , Acinetobacter , Coliforms.

6
Gram positive: Micrococcus , Bacillus , Arthrobacter , Lactobacillus , Streptococci.

c. Bacterial flora on the skin surface of freshwater fish :

Gram negative: Acinetobacter , Pseudomonas , Coliforms.

Grampositive: Micrococcus , Bacillus.

1.4.2 Factors that influence the growth of microorganisms

There are a number of external and internal factors (intrinsic to the substrate, that is, the foods in which they
grow) that affect the microbial growth.
Of the different external factors, temperature is the most important one.
(1) Thermophiles (high-temperature loving kind): growth optimum above
45 o C (35 o to 65 o C).
(2) Mesophiles (medium- temperature loving kind): growth optimum above
25o to 37 o C (15 o to 45 o C).
3) Psychrotrophs (cold-temperature tolerant kind): growth temperature is between 0 o - 25oC (optimum 15 o -
25 oC).
The bacteria on temperate water fish are all classified according to their growth temperature range as either
psychrotrophs or psychrophiles. Psychrotrophs (cold-tolerant) are bacteria capable of growth at 0oC but with
optimum around 25oC. Psychrophiles (cold-loving)are bacteria with maximum growth temperature around 20oC and
optimum temperature at 15oC (Morita, 1975). In warmer waters, higher numbers of mesophiles can be isolated. The
microflora on temperate water fish is dominated by psychrotrophic gram-negative rodshaped bacteria belonging to
the genera Pseudomonas, Moraxella, Acinetobacter, Shewanella and Flavobacterium. Members of the Vibrionaceae
(Vibrio and Photobacterium) and the Aeromonadaceae (Aeromonas spp.) are also common aquatic bacteria and
typical of the fish flora. Gram-positive organisms are Bacillus, Micrococcus, Clostridium, Lactobacillus and
Coryneforms can also be found in varying proportions, but in general, Gram-negative bacteria dominate the
microflora.

Interaction between growth factors

1) Microbial growth is arrested even when one of the growth factor is controlled. For example, bacterial
growth slows down under reduced pH or acidic conditions.
(2) If more than one factor becomes limiting, microbial growth is drastically curtailed or even stopped. For
example bacterial growth would stop under acidic conditions combined with low temperature and lower water
activity(aw).

Chapter 5: Spoilage of fish

1.5.1 Spoilage of fish
Spoilage of fish, post mortem, is mainly due to (1) oxidation, (2) autolysis and (3) bacteria. The major cause
of spoilage of fish is bacteria, particularly in the case of marine fishes.

The flesh and body fluids of newly caught fish are free from bacteria (except when the fish has bacterial
disease). The bacteria present on skin, adhering slime gills and intestine are normally saprophytic. Once the fish is
dead, these bacteria invade the fish tissue.There are three main routes for this attack.

1. From the gills into the flesh through the vasculatory (circulatory) system.
2. Through the skin by penetration.
3. Through the peritoneal lining (from the intestinal cavity).

Invation of bacteria through the first and second routes is faster. Entry through the peritonial lining can take
place only after perforation of stomach and intestinal walls, which normally takes longer time.

7
 The fish muscle contains 15 to 18% protein. Bacteria attack the protein and break it down to peptides and
amino acids. Initially, bacteria live and multiply in the fish tissue, utilizing the low molecular weight compounds like
carbohydrates and amino acids present in small quantities in the muscle. Due to post-mortem enzymatic break down
of the macromolecules in the muscle, enough low molecular weight compounds are formed, which serve as the food
of bacteria. Subsequently bacteria elaborate proteolytic enzymes, which break down proteins to peptone,
polypeptides, lower peptides and finally to amino acids. Bacteria will metabolize amino acids, in different ways leading
to the production of odouriferous and foul smelling compounds like ammonia, hydrogen sulphide, mercaptans, indole,
amines and organic acids.

When the fish is left in ambient temperature, which is usually 28.40C, tropical fishes get spoiled within 6 to 12 hrs.
depending on their size. In order to prevent such spoilage, many methods are in practice. Drying, icing, freezing and
use of chemicals are some of the usual methods. The basic principle involved in these methods of preservation of
fish is to control the activities of the microorganisms.
Typical spoilage compounds during spoilage of fresh fish Substrate and off-odour / off-flavour compounds
stored aerobically or packed in ice or at ambient produced by bacteria during spoilage of fish
temperature Substrate Compounds produced by
Specific spoilage organism Typical spoilage compounds bacterial action
Shewanella putrefaeciens TMA, H2S, CH3SH, (CH3)2S, Hx TMAO TMA
Photobacterium phosphoreum TMA, H2S Cysteine H2S
Pseudomonas spp. Ketones, aldehydes, esters, Methionine CH3SH, (CH3)2S
non-H2S Sulphides
Carbohydrates and Acetate, CO2, H2O
Vibrionaceae TMA, H2S lactate
Anaerobic spoilers NH3, acetic, butyric and Inosine, IMP Hypoxanthine
propionic acid
Amino-acids Esters, ketones, aldehydes
(glycine, serine,
leucine)
Unit 2: Chilling of fish
Amino-acids, urea NH3
Chapter 1: Fresh fish handling

2.1.1. Introduction to Icing

Icing is the most prevalent method of preserving fish. Ideal icing involves packing crushed ice and fish alternatively in
insulated boxes, in the fish to ice ratio of 1:1 (w/w).
about 2-3 hrs (the melting of the ice needs 80 calories of heat/g and this heat is removed from the fish in contact with
ice and hence, the fish get cooled). This lowering of temperature brings about

1. arrest of almost all enzymatic changes,

2. killing of about 50-60% of the mesophilic bacteria and
3. slowing down of the activities and growth of all other bacteria, which are cold-loving (psychrophilic) and
cold-tolerant (psychrotrophic).

As a combined effect of all these three factors, the spoilage of fish is delayed to a considerable length of time in
ice. During iced storage of fish, there is an initial drop of bacterial count due to the death of the cold sensitive
mesophiles. The surviving cold tolerant bacteria, however, get adapted to growth in low temperature. Consequently,
there is a gradual increase in population, which takes about 6 to 8 days to reach a count of one million per gram or
above. By that time, the fish has reached the stage of incipient spoilage.
Qualitatively, there is a selection of bacterial flora during iced storage of fish. Irrespective of the composition of the
initial flora, the Pseudomonas/ Alteromonas group emerges as the predominant group of bacteria at the time of
spoilage. This is because most of the psychrotrophic bacteria capable of spoilage belong to these genera.

8
 In the case of tropical fishes, it is not the psychrophiles, but the psychrotrophs, which are the actual spoilers during
iced storage. These psychrotrophs, whose population is very low in the fresh tropical fish, easily adopt to grow at
low temperature during iced storage and grow very rapidly and spoil the fish. Further, psychrotrophs have a shorter
generation time compared with psychrophiles.
2.1.2. Advantages of chilling of fish with ice
i) Temperature reduction. It reduces the temperature to about 0oC. The growth of spoilage and pathogenic micro-
organisms is reduced, thus reducing the spoilage rate.
ii) Melting ice keeps fish moist. This action mainly prevents surface dehydration and reduces weight losses. Melting
water also increases the heat transport between fish and ice surfaces (water conducts heat better than air): the
quickest practical chilling rate is obtained in a slurry of water and ice (e.g., the CSW system).
iii) Advantageous physical properties. Ice has some advantages when compared with other cooling methods, including
refrigeration by air. The properties can be listed as follows:
a) Ice has a large cooling capacity.The latent heat of fusion of ice is about 80 Kcal/kg. This means that a
comparatively small amount of ice will be needed to cool 1 kg of fish.
b) Ice melting is a self-contained temperature control system. Ice melting is a change in the physical state of ice
(from solid to liquid), and it occurs at a constant temperature (0oC).
iv) Convenience. Ice has a number of practical properties that makes its use advantageous. They are:
a) It is a potable cooling method. It can be easily stored, transported and used. Depending on the type of ice, it can
be distributed uniformly around fish.
b) Raw material to produce ice is widely available. Although clean, pure water is becoming increasingly difficult to
find, it is still possible to consider it a widely available raw material and it should be properly treated, e.g.,
chlorination.
c) Ice can be a relatively cheap method of preserving fish. This is particularly true if ice is properly produced (avoiding
wastage of energy at ice plant level), stored (to avoid losses) and utilized properly (not wasted).
d) Ice is a safe food grade substance. If produced properly utilizing drinking water, ice is a safe food substance and
does not cause any harm either to consumers or those handling it. Ice should be handled as food.
v) Extended shelf life.
The overall reason for icing fish is to extend shelf life fresh fish of in a relatively simple way as compared to storage
of un-iced fish at ambient temperatures above 0oC. Extension of shelf life means producing safe fresh fish of acceptable
quality.
2.1.3. Chilled storage
In general, two types of plastic fish boxes are used: stack-only and nest/stack boxes.

1. The handling rate is necessary to prevent quality loss because of delayed icing. Pre-chilling can be of advantage
to compensate lack in handling rate.
2. Handling methods, which make it possible to guarantee that the icing procedure is sufficient to chill the fish
to 0oC and maintain this temperature until landing.
3. The hold must be constructed in such a way, that safe and easy stacking of the boxes can take place.
4. Hold insulation of a relatively high quality should be considered. A small mechanical refrigeration plant can be
of advantage. Air temperature in the hold should be +1o or -3oC.

Use of antimicrobial agents in ice and cold liquids

By ordinary method of icing, fish can be preserved for 10-12 days. However, studies have shown that use of
antimicrobial agents in ice or cold liquids, enhance the shelf life of fish by 60 to 100%. The following antimicrobial
agents were used.

a) Chlorine compounds

If liquid chlorine or bleaching powder is mixed with ice or cold liquids, shelf life of fish can be enhanced. If the cooling
media (ice or chilled liquid) contains less than 30 ppm of chlorine, no damage is done to fish. However if the proportion
is more than this, the fish gets acrid smell. Also due to oxidation, the colour of fish may change. By this method fish
can be preserved for 18-20 days.

9
b)Antibiotics

By the use of antibiotics in ice or cold liquids, shelf life of chilled fish can be increased. Among the antibiotics,
chlorotetracycline and oxytetracycline are important. These are powerful broad spectrum antibiotics and are very
effective even at low concentrations. Antibiotics were used for chilling fish as follows.

1. By dip treatment in antibiotic solution and then chilling.

2. Incorporating antibiotic in water and then ice is prepared and used for chilling.
3. Incorporating in chilling liquids (CSW or RSW)
4. It has been found that dip treatment of fish in water containing 30 to 100 mg of CTC for 1-5 min and then icing
increases the shelf life by 5-10 days. Ice containing 5 mg/kg of CTC, when used, increase the shelf life of fish
from 10 days to 20-22 days. However at present use of antibiotics in banned as per Codex Alimentarius
standards.
2.1.4 Storage method

There are three methods of storing fish in ice on fishing vessels

1. Bulking:

The fish hold is a insulated chamber located at the near

side of the Boat. It is divided into different compartments
using wooden boards supported by an upright beam. A
layer of ice at least 5 cm thick is spread over the bottom of
a compartment followed by a layer of fish. Ice is then
spread over the fish and around the edges so that the fish
are not in direct contact with the sides of the board.

Further layers of fish and ice are added until a depth of

about 45cm ice and fish is achieved, with a layer of 5 cm
of ice at the top. A horizontal wooden board is now placed over the section. The wooden board must be supported
by the stanchion structures, not by the fish and ice in the lower compartment. More fish and ice are added in the
same way, again to a depth of 45cm. The operation is repeated until the compartment is full. Wooden boards and
stanchions must be kept clean and out of direct contact with the fish.

2. Shelving:

The fish hold is divided into sections as it is for bulking but this time removable shelves spaced at about 23 cm are
used for holding the fish. The lowest shelf is covered with a layer of at least 5 cm of ice. Fish are placed in rows on the
ice and more ice is used to cover the fish to about 5 cm. Only one layer of fish is to be put on to each shelf. Shelves
must be supported by stanchions, not by the fish and ice below.

Boxing: Fish boxes come in a variety of sizes and materials. Ideally, a

box should:
i. Be strong and robust.
ii. Be able to be stacked so that the weight of the top boxes
are taken by the boxes below, not by the fish in the box
below.
iii. Be able to nest to save on stowage space when empty.
iv. Be easily cleaned and, if necessary, sterilized.
v. Allow ice melt-water to flow away outside the box below
and not through it on to the fish in the lower box.
vi. Have good thermal insulation.

10
Fish boxes should be used as follows
A layer of ice 5 cm thick should be placed in the bottom of the box, followed by a layer of fish. A thin layer
of ice follows, interlacing fish and ice, until the box is almost full. The ice should be placed around the sides of the box
as well as amongst the fish and the top layer of fish should be covered with at least 5 cm of ice. The box must not be
over filled. This prevents crushing the fish when the boxes are stacked into the fish hold. When boxing fish, the fish
hold does not require any compartments or Stanchions.

Refrigerated fish rooms

Some fish rooms not only have insulation but also have refrigeration facilities. Earlier this refrigeration is
usually in the form of cooled grids and pipes on the roof of the fish room. Now-a-days a unit cooler with fan is placed
in the chill room. It is important to get the maximum cooling effect from the ice that is allowed to melt. For this reason
the temperature in the fish room should not fall below 0oC. In order to prevent melting of ice, it is common practice
for refrigeration to be used only before fishing starts when the vessel is carrying ice alone. Under these circumstances,
the temperature can be below 0oC.

Insulated boxes
In small boat fisheries, insulated boxes are used to carry ice to sea, and for storing ice and fish, when fishing
occupies only a short period. The size of the boxes depends on the size of the boat and the amount of fish normally
caught in a day’s fishing. The ambient temperature will govern the amount of insulation required in the box, though
about 10-15cm of expanded polystyrene is common. Insulated boxes for small fishing boats can often be made
locally at low cost.
Chapter 2: Calcultation of the ice requirement for cooling fish

2.2.1.Calculation of the ice requirement of cooling fish

The mass of ice needed to cool fish from the initial temperature to the final holding temperature can be
calculated from an expression, which equates the heat taken up by the ice with the heat lost by the fish.

Heat taken up by Ice = Heat lost by fish

(Mi) (Li) = (Mf) (Cpf) (ti-tf) ………….(1)

Where

Mi = mass of ice which melts (kg)

Li = latent heat of fusion of ice (80 kcal/kg)
Mf = mass of fish (kg)
Cpf = specific heat of fish (kcal/kgoC)
ti = initial temperature of fish (oC)
tf = final temperature of fish (0oC)
From equation (1) the ice requirement will therefore be:

(Mf) (Cpf) (ti−tf)

Mi = ……….(2)
(Li)

The specific heat of lean fish is 0.8 kcal/kg oC and this value should be used if there is a species mix or if there
is a possibility that all the fish are of a lean species.

The specific heat value, however, may be refined to take account of variations in the oil content of the fish and
this refined value may be used if the fish composition is reasonably consistent.

Cpf = 0.5 Xl + 0.3 Xs + 1.0 Xw (3)

11
Where Cpf = specific heat of fish (kcal/kg)
Xl = mass fraction of lipids (oil)
Xs = mass fraction of solids
Xw = mass fraction of water
To illustrate the effect of oil content on the quantity of ice required for chilling the following comparison is
made between lean and fatty fish.

Example (1) – 100 kg lean fish with 1% lipids, 19% solids and 80% water at an initial temperature of 20oC:

Cpf = (0.5 x 0.01) + (0.3 x 0.19) + (1.0 x 0.8) = 0.862 kcal/kg oC

100 x 0.862 x (20−0)

Mi = = 21.55 kg of ice
80

Example (2) – 100 kg of fatty fish with 21% lipids, 19% solids and 60% water at an initial temperature of 20oC.

Cpf = (0.5 x 0.21) + (0.3 x 0.19) + (1.0 x 0.6) = 0.762 kcal/kgoC

100 x 0.762 x (20−0)

Mi = = 19.05 kg of ice
80

Since the calculation for fatty fish shows only a small reduction in ice requirement and, with most species the
oil content is variable, it is advisable to treat all fish as lean fish.

2.2.1.1. Heat Requirements

The heat required to change from a solid to a liquid is known as the
Ice About 0.5
latent heat; 1 kg of ice requires 80 kilocalories (kcal) of heat to melt it. This
figure of 80 kcal/kg is known as the latent heat of fusion. It is this property Wet fish About 0.96 (usually
of requiring a large amount of heat to melt ice that makes it such a good taken as 1)
cooling agent. One kilocaloryis the amount of heat required to raise the Frozen fish About 0.4
temperature of 1kg water by 1 C. o Air About 0.25
More heat is required to warm water than almost any other Most metals About 0.1
substance. This capacity of substances to hold heat, when compared to
water, is known as the specific heat. Specific heat of water is 1, for other substances it is less than 1.

Specific heat can be used to discover how much heat has to be removed to cool a substance, i.e.
Heat to be removed = weight of substance x temperature change x the specific heat,
To cool 60 kg ice from – 5 to 10oC requires the removal of:
60 x [ - 5 – (-10)]oC x 0.5 (sp. Heat of ice) = 150 kcal

We can now calculate how much ice is needed theoretically to cool a given weight of fish:
If we want to cool 10 kg fish from 25 to 0oC, we would need to remove
10 x 25 x 1 = 250 kcal
But when ice melts it adsorbs 80kcal/kg.
Thus the weight of ice required = 250 / 80= 3.12 kg.

This is strictly a theoretical calculation and does not take the following factors consideration

1. Ice is also melted by the surrounding air; thus a lot of ice is lost, particularly at high ambient temperatures,
unless the fish and ice are protected from the ambient heat, preferably with insulating materials.
2. How the fish are packed in ice.
3. The length of time that the fish needs to be kept chilled once cooled.
4. How quickly the fish are chilled.

12
 2.2.1.2. Calculation of the ice requirement for the storage of fish
Even if you are concerned with only one batch of fish held in identical containers, there are likely to be variations
in ice melting rates, which make it difficult to calculate the ice requirement accurately. For instance, the
containers are stacked, and then ice melting may be different in containers located at the top, bottom, sides and
within the stack.
All containers should be treated equally and the assumption made that each container is fully
exposed to the surrounding air.
As a first step, heat transfer may be calculated using the following simple expression:

q = A.U. (to – tc) kcal/day (4)

Where q = heat entering the container (kcal/day)

A = surface area of the container (m2)

U = overall heat transfer coefficient (kcal/day m2 oC)

to = temperature outside the container (oC)

tc = temperature inside the container (oC)

This overall calculation of heat transfer may require to be done in parts, for instance, the lid or base of
the container is made of a different material or has a different thickness. The heat transfer through the
various areas are then added together to give the total heat transfer.
The heat entering melts the ice, therefore it follows that:

q = Li . mi (kcal/day) (5)

Where q = heat required to melt ice (kcal/day)

Li = latent heat of fusion of ice (usually taken as 80 kcal/kg)
mi = mass of ice melted (kg/day)
In order to develop a mathematical expression for ice melting rate during the storage period it is assumed
that ice melting inside the containers is only due to heat transferred from the surrounding air. In this steady state
approach, quantities (4) and melting (5) should be equal, therefore it follows that:

Li . mi = A.U. (to-tc) (6)

Thus, the ice requirement will be:

mi = A.U. (to-tc) / Li = kg/day -------------------- (7)

To include the element of ice melting due to radiated heat will make the calculation extremely difficult.
Therefore, if the containers cannot be protected from direct sunlight or any other radiated heat source during the
storage period, the calculated values for the ice requirement should be upgraded.

Chapter 3: Manufacturing of different types of Ice

2.3.1. Introduction
There are many methods of producing ice. The manufacturing process of one may involve efficiencies or
advantages over another or the ice produced by one process may be better suited to the intended application than
another. Commonly used ices are block ice, flake, plate and tube ice.

13
 2.3.1.1. Block Ice
In the manufacture of block ice, tapered
rectangular cans filled with potable water are
immersed in a tank of refrigerated brine solution .

The brine solution which is cooled to about –

o
30 C by a refrigeration process extracts the heat
from the water and produce block ice within the
can. The cans are then removed from the tank and
thawed for a short time in a tank of water to release
the block from the can. The blocks are then stored in
a chill room and can be crushed on demand. The
freezing period is typically between is 16 and 24
hours although plants known as rapid blocks are
available that have freezing period of only a few
hours. The quick freeze is achieved by the direct evaporation of the refrigerant in a jacketed mould fitted with finger
evaporators. The blocks are released by a hot gas defrost and are sub-cooled below 0°C. Rapid block plant requires
less floor space than brine tank system.
2.3.1.2. Flake Ice
A thin sheet of ice, 2 to 3 mm thick, is formed by
spraying water on to a refrigerated drum; it is scraped
off to form dry sub-cooled flakes. In some models, the
drum is rotated; in others, the scraper or knife is
rotated; in most designs, the drum is vertical. Flake ice
production is continuous and automatic; ice is formed
within a few minutes of starting up the machine, which
can be adjusted so that the thickness of the ice
produced can be varied.

The ice leaving the drum is cooled below 0oC

(sub-cooled) and is stored in a refrigerated silo, usually
immediately below the ice-making machine. Flake ice is ready for use immediately it leaves the refrigerated surface.
It is important that the ice shall be kept sub cooled condition, if it is not, then regelation can occur so that the ice forms
a solid mass which is very difficult to handle.

14
 2.3.1.3. Plate Ice
In the manufacture of plate ice, water
is sprayed and frozen on to the outer
surface of refrigerated plate to form a
sheet of ice, which usually released by
an internal hot gas to defrost.

Thickness of the ice can be

varied with in limits, by alteration of
the defrost cycle, but it is commonly
get between 8 and 15mm. As the ice
falls it is chopped by a rotating cutter
and this process is automatic.

2.3.1.4.Tube Ice

Tube ice is formed a vertical shell and tube vessel by passing

water inside the tubes which are cooled by the circulation of
refrigerant on the outer surfaces.

When the ice reaches the desired thickness the water flow is
stopped automatically. The refrigerant is removed to a surge
drum and hot gas circulated around the tubes causing the ice
tubes of melts and slide down. As the ice falls from the tubes
a rotating knife blade breaks it to desired size. The ice tubes
are about 50mm in diameter with a fall thickness of 10-12mm.

Physical characteristics of ice utilized in chilling fish

Types Approximate Dimensions Specific (m3/t) Volume Specific Weight

(t/m3)
Flake 10/20-2/3 mm 2.2-2.3 0.45-0.43
Plate 30/50-8/15 mm 1.7-1.8 0.59-0.55
Tube 50/D-10/12 mm 1.6-2.0 0.62-0.5
Block Variable 1.08 0.92
Crushed Variable 1.4-1.5 0.71-0.66
block

Properties of water and ice

Properties Metric units

Pure water Density at 15oC 1 kg/l 1/m 3

Specific heat 1.0 kcal/kgoC

Latent heat of fusion 80 kcal/kg
Thermal conductivity (at 10oC) 0.5 kcal/mboC
Freezing point 0oC
Boiling point 100oC
Sea water density 1.027 kg/l,1.027 t/m3

15
 Specific heat 0.94 kcal/kgoc,0.93 kcal/kgoc
Freezing point at salinity of
-0.6oC
1.0%
-1.2 oC
2.0%
-1.6 oC
3.0%
-1.9oC
3.5%
-2.2 oC
4.0%

Chapter 4: Super chilling (0ºC to - 4ºC)

2.4.1. Introduction
Storage of fish at temperatures between 0oC and –4oC is called super chilling or partial freezing. The shelf life
of various fish and shellfish can be extended by storage at sub-zero temperatures. Sub-zero storage temperatures in
fishing vessels can also be obtained in refrigerated sea water (RSW) where the freezing point of water is reduced by
NaCl or refrigerant.
Compared to ice storage, the RSW systems chill fish more rapidly, reduce the exposure to oxygen, reduce the pressures
that often occur when fish are iced and also give significant labour-saving. Super chilling extends product shelf life, but
a negative effect on freshness/prime quality has been observed for some fish species. Merritt (1965) found that cod
stored at –2oC for 10 days had an appearance and texture inferior to fish stored at 0oC in ice. The drip of the super
chilled fish was increased and at –3oC the texture of whole cod made them unsuitable for filleting. RSW storage gives
several fish species a salty taste due to the up take of salt from sea water.
2.4.1.1. Refrigerated Sea Water (RSW)
Seawater has a salt content of around 3 to 3.5%. At 3.5% salt, seawater has a freezing point of about –2oC.
Thus, if seawater is refrigerated, it is possible to reduce the temperature so that a storage temperature of –1oC can be
achieved. The most important advantage of RSW over icing is the ease of handling and storage onboard, with a
resultant saving of labour. In a purse - seiner, the fish can be brailed from the net direct into the tank and brailed out
of the tank at the landing point.
Pumping systems have been developed for unloading fish as large as 10kg each at a rate of more than 25
tonnes/hour. In the most sophisticated systems, the vessel can be equipped with a pump which pumps the fish out of
the sea into holding tank and the same pumps can be used for unloading.
RSW is popular on large vessels, while CSW can be applied even on small boats. For RSW, larger capital
investment is required for the mechanical refrigeration system. In order to avoid inefficient, too slow chilling of the
catch, not more than 800kg of fish should be packed per kilo litre of the brine. RSW may also be used to cool the catch
by spraying over the top surface of ungutted fish or shrimp in the hold. Rapid chilling and holding of the catch in RSW
is a popular practice on board large vessels which catch shrimps.
Chilling of the catch in RSW before freezing on board freezer trawlers is a very effective method of
preventing rapid quality loss of the fish on subtropical and tropical grounds. The shelf life and quality seafoods chilled
or stored in CSW or RSW depends upon the species characteristics and the condition of chilling.
Generally, it is a few days longer than that of iced fish. Some species are vulnerable to loss of appearance
due to bleaching or leaching. The uptake of salt may significantly decrease the market value of some seafoods, like
shrimps, snappers. The advantages of RSW are

1. Labour saving in stowage

2. Rapid chilling, 2 to 3 times as fast as with ice
3. Elimination of crushing and ice pitting
4. Lower chill temperatures
5. Washing effect
6. Ease of unloading

16
 2.4.1.2. Chilled sea water (CSW)
It is also possible to cool seawater by mixing ice with it. These systems are usually referred to as chilled seawater
(CSW) and can be extremely simple. A mixture of seawater and ice, usually 1:1 to 1:2 on a volume basis at the start,
forms an ice slurry that has a temperature of about –1.5oC. In such ice slurry or chilled seawater (CSW), the heat
transfer between the fish and the cold medium occurs by convection. Thus, the rate of chilling is higher than in ice. Ice
and salt should be added to the slurry during chilling to compensate for the loss of ice due to melting and to maintain
the salt concentration at about 3%. Agitation in tank prevents accumulation of the ice at the surface and formation of
a large temperature difference between the upper and bottom part of the container. Furthermore, forced convection
increases the rate of heat transfer.
The rate of chilling of fish in CSW is especially high in the range from ambient temperature to about 5oC, while
below 5oC, it is not significantly higher than that in crushed ice. Thus, CSW is often used only for rapid initial chilling of
highly valued fish onboard. Another use of CSW is for chilling and holding the catch onboard in insulated containers.
The shelf life of ungutted fish kept in CSW containers is a few days longer than that of fish carefully chilled with flake
ice. Furthermore, no textural damage occurs except for some loss of scales. In the case of small fatty fish, the CSW
treatment offers some protection against rancidity. Compressed air or nitrogen is injected into the bottom of the tank
to ensure rapid and even cooling, the gas flow rate should be 2-4kg/h at a pressure of about 35kN/m2. The contents
should be agitated at least for 6 hours.
The recommended ratio of fish to seawater is between 3:1 to 4:1.
In RSW systems, it is usually necessary to provide pumped circulation of the water to ensure even mixing. This is
sometimes done with CSW systems and the motion of the vessel providing adequate mixing. A rough calculation of
the amount of ice needed to provide adequate cooling in a CSW system can be made quite easily in the following
manner:
Suppose 8 tonnes of fish (8,000 kg) are to be cooled from 24 to –1oC and 4:1 ratio of fish to seawater is used, the
mixture of seawater and fish would weigh 10 tonnes (10,000 kg) and the amount of heat to be removed would be:
10,000 x 25 x 1 (taking 1 to be the specific heat of both fish and water) = 2,50,000 kcal
But, when ice melts, it absorbs 80 kcal, thus the weight of ice required:
2,50,000 /80 = 3,125 kg

Thus, just over 3 tonnes of ice would be required to provide the initial cooling. This would only reduce the
temperature of the fish and would not be sufficient to maintain the lower temperature.
Additional ice would be required to provide against heat leak. The amount of additional ice needed would
depend on the efficiency of the insulation of the tank or fish room, the outside ambient temperature and the
duration of the trip.
As a guide it may be noted that if the same 10 tonnes mixture of fish and seawater is to be cooled from 24 to
o
1 C, it would impose a load of 1,000 mega joules on the refrigeration system. If this amount of heat is to be removed
in about 6 hours, a plant will have to be installed with a power requirement of about 12 kw for the compressors.
2.4.1.3. Slurry Ice in Fish Preservation
The introduction of Deep Chill ice for direct contact cooling of fish made significant improvements in cooling
and handling processes over other forms of ice. These systems have daily ice capacities ranging from 7 to 200 mt.
Deep Chill ice
The Deep Chill ice consists of small crystals, either in dry form or pumpable slurry, and has the following beneficial
effects:
 provides the best chilling of fish compared to other types of ice while avoiding freezing
 the cooling process is maintained at a constant level as the ice melts
 prevents fish from bruising, because of the soft and flexible nature of ice
 easy to mix, handle and transport due to pumpability of ice slurry
 possibility of brine drainage to provide almost salt free ice for extended storage of fish

2.4.1.4. Cooling and Storage Tests

While studying the chilling effect of Deep Chill ice slurry with flake ice, it was found that, ice slurry provides more
efficient chilling than flake ice. This is because ice slurry effectively eliminates hot spots in the fish container and
provides maximum contact of ice surrounding the fish.

17
 1. It significantly retards bacterial growth and reduces fish tissue degradation.
2. The temperature drop in the fish body is extremely significant in the early stage of the cooling process.
3. The final equilibrium temperature of the fish is approximately 10C lower, using the same amount of ice slurry
compared to flake ice.
4. The difference in cooling level occurs due to the larger enthalpy change of the ice slurry. No uptake of salt was
detected while using slurry with 0.8% salinity.

The quality of the product, such as the texture, colour and bacteria retardation is greatly improved. Automated fish
handling with ice slurry improves the productivity of the plant, and the plant is able to deal with higher load without
incurring too many shifts per day.

Chapter 5: Application of ozone / chlorine in seafood processing

2.5.1. Application of ozone in seafood processing
Pathogenic bacteria and viruses present in fish are highly sensitive to residual ozone in water. 99.9% or higher
inactivation of Aeromonas spp., Vibrio anguillarum and Yersinia ruckeri is obtained at residual ozone concentrations
of 0.15 – 0.20 mg / l within 60 seconds in natural lake, brackish and sea water. In fish farm effluent, a residual
concentration of 0.3-0.4 mg/l is required for Aeromonas inactivation. Among fish pathogenic viruses, high sensitivity
toward ozone has generally been reported. This also applies to viruses with high UV resistance, i.e. IPNV and WSBV as
reported by many researchers.
Because of its instability, ozone must be generated and used on-site. The most efficient method is by the electric
discharge technique, which involves the passage of oxygen gas, or air, across the gap of narrowly spaced electrodes
under high voltage. Due to the energy costs of producing ozone, it is important to optimize the transfer efficiency from
gas to liquid phase.
Ozone in seafood processing and fisheries
The method of use of ozone started in late 1996 onboard purse - seiners with an idea to decrease bacteriological
problems in the circulation of RSW system.
Today, ozone is used in the seafood processing and fishery industry in areas such as:

 Purse-seiners – seawater is ozonised and circulated through RSW tanks and circulation systems so that it
comes in contact with every surface and carries out clearing.
 Well boats – when transporting live fish you have to obtain a high level of hygiene in order to prevent mortality
by pathogens. By using Ozonised Sea water to reduce contaminants from tanks and circulation systems, and
disinfects them in accordance with applicable requirements.
 Processing plants – ozonised sea/fresh water is used for cleaning and disinfection of process area, equipment
and pipelines.
 Inlet water for fish hatcheries
 Waste water treatment – effluent from aquaculture, fish processing and fish slaughtering. The waste water
treatment systems must be tailor-made according to regulations and are based on a combination of methods
such as filtration, flotation and separation, and ozonisation.
 Odour-ozone has been successfully used in several applications to treat odour from fish or waste storage.

2.5.2. Use of Chlorine in Fish Processing

 Chlorine is used in the fish processing sector as a water disinfectant and is probably the most
widespread disinfectant in use. Its uses include washing fishery products, addition to water for making ice for
chilling fish, and in water for thawing frozen products. It is also used in water to cool canned fish after retorting
to prevent "leaker” spoilage. Chlorine is commercially available in several forms, the most common being a
granular or powdered form as calcium hypochlorite or in liquid form as sodium hypochlorite (NaOCl) or
bleach. In any of these forms it acts as a powerful oxidizing agent and reacts with a wide variety of
compounds. Chlorine is a gas under normal pressure and temperature. It can be compressed into a liquid and
distributed in cylinders and fed automatically into water supplies of fish processing plants or on-board fishing
vessels.
 The use of chlorine dioxide is less common in fish processing. Some of the reported advantages of
chlorine dioxide over aqueous chlorine as a disinfection agent are that, it is seven times more potent than
aqueous chlorine in killing bacteria; the bactericidal activity of chlorine dioxide is not affected by alkaline
18
 conditions and/or the presence of high levels of organic matter. For these reasons, it is under investigation
by many authorities for use in the fish processing sector.
2.5.2.1.Concentration of chlorine used in Fish processing
 It has been established in the fish processing industry that the injection of chlorine into a supply cold water
used for general wash-up helps to control microbial contamination. Chlorine dosage should be around 10
ppm during the normal use and 100 ppm of residual concentration during the clean – up.
 Codex recognize that up to 10 mg/l active chlorine in fish processing water and ice that contacts seafood
to be generally recognized as safe (GRAS).
 An on-line hypochlorite injection system recommended for seawater treatment is a dosage of 20ppm. This
system, together with improved handling and storage of cooked shrimp, has been found to double the
chilled storage life while improving both the eating and microbiological quality of products. Concentrations
of 20-30 mg/L in running water have been used to reduce the number of Listeria monocytogenes, while
thawing frozen salmon. The removal of this pathogen from the surface of fish destined for cold smoking is a
major health gain for the industry as the temperature of the cold smoking process are insufficient to kill L
monocytogenes. Chlorine at levels of 20-25 mg/L were shown to be effective in killing strains of both
Escherichia coli and L monocytogenes in a fish model system under laboratory conditions.
2.5.2.2. Marginal Chlorination
 There are two important chlorination methods: marginal chlorination and super-chlorination with or without
dechlorination. The latter includes break-point chlorination and in-plant chlorination.
 Marginal chlorination is the addition of just sufficient chlorine to water to produce a residual level. The
residual chlorine may be either free or combined depending on the amount of ammonia present in the
water. This level of chlorination usually destroys pathogens.
2.5.2.3. Super Chlorination
 The chlorine dosage applied is much greater than that of marginal chlorination. The chlorine dosage depends
on the character of the water and the purpose for which the water is used. A considerable amount of
chlorine may be present after a given contact period. Therefore, for certain purposes, the water is
dechlorinated. Super chlorination includes break-point chlorination and in-plant chlorination.
2.5.2.3.1. Break-point chlorination

When small amounts of chlorine are added to water, first, chlorine reacts with certain impurities present in the
water. These impurities are substances responsible for chlorine demand. Chlorine reacts with nitrogenous
compounds to form chloramines or other chloronitrogen compounds. On further addition of chlorine, a free residual
chlorine appears. On still further addition, the free residual chlorine gradually increases until it reaches a particular
concentration. The concentration of free chlorine is determined by the physical and chemical nature of the water.
When the level of free chlorine is increased beyond this concentration, oxidation reaction occurs between free
chlorine and chloronitrogen compounds.

The level of the free residual chlorine is decreased during the oxidation of chloronitrogen compounds. When the
oxidation is completed, the addition of chlorine to water results in corresponding increase in chlorine concentration.
The point after the first rise in concentration at which the free residual reaches its lowest level is known as the
breakpoint.

The advantages of break point chlorination are the following:

 Complete oxidation of ammonia and other compounds in the water

 Correction of tastes and odours of biological origin
 Correction of tastes and odours due to phenol and other substances
 Reduction of colour due to organic matter
 Improvement of bacterial quality of water

19
 2.5.2.3.2. In-plant Chlorination
 In-plant chlorination has been defined as break-point chlorination of all water as it enters the plant to a
degree where a good persisting residual chlorine occurs. In-plant chlorination of processing water is
employed in many food industries.
 Chlorine gas is generally considered the best source for in-plant chlorination where large volumes of
water are to be chlorinated. But for chlorination by chlorine gas, a chlorination equipment is needed.
Hypochlorites are a good source when only small amounts of chlorine are required. Since seafood processing
industry requires only small amounts of chlorine, hypochlorites can be used by the industry.

Unit 3: Freezing preservation of fish

Chapter 1: Principle of freezing
 Heat Units
3.1.1. Specific heat
 Specific heat is the quantity of heat that must be supplied or removed from a gram of substance to bring about
change in temp of 1ºC. It is expressed in terms of kilojoules (KJ) or kcal in applied science.
 Heat capacity/specific heat of fish depends on its chemical composition; that is the content of water, fat and
minerals. It is the total heat capacity of individual components which is given by the following formula.
 Cf = Cw X W+L+Cm X m
Where,
Cf= heat capacity of fish (kcal/kg)
Cw = specific heat of water (kcal/kg)
CL = specific heat of fat/lipid (kg)
Cm = specific heat of minerals (kg)
W, L and M are contents of water, lipid and mineral in Kgs in a kg of fish.
Example : Specific heat of fish also depends on temperature. There is no applicable change in specific heat. When
fish is cooled from 30ºC to 0ºC (prior to freezing)and in this range of temperature, specific heat of major
components of fish are given as follows.

Water - 4.18 KJ/kgºC or 1 kcal/kg ºC

Fat - 2.075 KJ/kgºC or 0.5 kcal/kg ºC
Dry matter other than fat - 1.505 KJ/ kg ºC or 0.36 Kcal/ kg ºC (i.e., protein + minerals)

Based on these values specific heat of various fishes were calculated and found to be between 0.74 to
0.91 Kcal / kg ºC between 30 to 0ºC. As fish gets cooled further its specific heat goes on decreasing. Chilled fish
has sp. heat of 0.845 kcal/ kg ºC which gets reduced to 0.440 kcal/ kg ºC, when the same fish is frozen. This indicates
less quantity of heat is required to change the temperature by 1ºC in frozen fish when compared to unfrozen fish.
This difference is large especially when all the water in the fish is frozen (i.e., to below -8ºC).

3.1.2. Thermal conductivity

 In an unevenly heat body, where there is always a difference in temperature, there is always a flow of
heat from warmer to colder layers. This type of heat exchange depends on direct heat transfer between
adjoining particles and this continues until the temperature of body is uniform. The rate of heat transfer
depends on the co-efficient of thermal conductivity of the substance which is defined as thermal
conductivity of medium in which a unit of heat (calorie) flows through a unit surface area of unit thickness
at a temperature gradient of 1 unit. Consequently expresses the amount of heat (calories) passing through
a unit area of flat surface having unit thickness at a temp difference of 1 unit in unit time. The units of thermal
conductivity are erg/cm. sec. ºC or cal/cm sec, ºC or kcal/m. Lr. ºC.
 When fish is heated or cooled, the rate of conduction of heat within the fish or rate of removal of heat
depends on the co-efficient of thermal conductivity. It depends on chemical composition of fish, physical
structure, specific gravity, specific heat and temperature of fish. The thermal conductivity in fish is important
it determines the chilling, freezing and thawing process in it. Effect of temperature on the co-efficient of their
mass conductivity can be illustrated by the following table.

20
 Temp ºC Co-efficient of thermal
conductivity
+30°C 0.4 kcal/m.hr. ºC
0º C 0.401-405 kcal/m.hr. ºC
-1 ºC 0.522 kcal/m.hr. ºC
-2 ºC 0.638 kcal/m.hr. ºC
-4.5 ºC 0.94 kcal/m.hr. ºC
-30 ºC 1.5 kcal/m.hr. ºC

 Co-efficient of thermal conductivity in fish can be theoretically calculated if its water and dry matter, content
is known . lf=lo x XW +ln X N
lf = coefficient of thermal conductivity of fish
lo = coefficient of thermal conductivity of water in fish
ln = coefficient of thermal conductivity of dry matter in fish ( 0.22 kcal/m.hr.ºC)
W and N are respective water and dry matter %age in fish.
 Co-efficient of thermal conductivity of ice is 2.07 kcal/m.hr.ºC which is almost 4 times that of water (0.52
kcal/m.hr. ºC). Hence heat transfer in frozen fish is faster than unfrozen chilled fish.

3.1.4 Latent heat

 It is the quantity of heat required to change the state or condition or phase under which a substance exists
without change in temp. Eg. a definite quantity of heat must be transferred to ice at OOC to change it to water
at the same temperature. This definite quantity of heat is known as latent heat of fusion of water/ice.
 Latent Heat of fusion of ice/water = 79.7 cal/g or 80 cal/g
 It is also expressed in terms of BTU/lb
 1 BTU/lb = 0.5556 kcal/kg or/k cal/kg = 1.80 BTU/lb
 Latent heat of fusion of any food can be calculated if its water content is known.
 Latent heat = 80 X % water in food/100
 Latent heat of fusion of some food stuffs.

Food Product Latent Heat of fusion

BTU/lb kcal/kg
Eggs 100 55.56
Fish 101 56
Green peas 106 58.88
Milk 124 68.85
Oyster meat 125 69.44
Pork 86 47.77
Water 144 80

3.1.5 Principles of freezing

It is a common fact that the foods with high moisture content rapidly deteriorate in quality. During freezing, the water
in the food is separated out from other food components, and is frozen. Thus, a food is protected, preserved from
deteriorating influences such as temperature and water. The lower temperature slows down the rate of chemical
reaction and water is also removed from the sphere of activity. The water is reabsorbed by the food components upon
thawing and food is restored to its original quality. However, when a food is frozen, ie. When the water undergoes
transition from liquid to solid state and also in subsequent thawing ie., when there is transition from solid to liquid
state, there is a damaging effect on food. The texture and the structure of the food may be affected to such an extent
that it may become irreversible and upon thawing the food will not be restored to its original condition.

21
 Fresh fish flesh normally contains 60-80% water and the freezing process converts most of this water into
ice. At normal atmospheric pressure, pure water will change from liquid to solid (ice) at 0 oC, i.e., it will freeze. The
water in fish flesh contains salts and chemicals, which have the effect of lowering the temperature at which the water
begins to freeze. The water in fish flesh begins to freeze at about –1oC and as the temperature drops below –1oC, more
water is frozen out and the concentration of salts in the remaining water rises, so that its freezing point is lowered
further. At5oC, it would appear that all the water is frozen, but over 20% of the water in fish muscle is still unfrozen.
Even at –30oC, approximately 10% of the water remains unfrozen.The above defined heat units are applied here.
In order to change the physical state of a substance from a liquid to a solid, energy or latent heat has to be
removed from the substance. In order to lower the temperature of 1g of water by 1oC, at temperatures above 0oC,
1 calorie of heat must be removed this is known as the “specific heat”. However, to change water at 0oC to ice at 0oC,
80 calories must be removed for each g of water. In other words, the specific heat of liquid water is 1 and the latent
heat of changing liquid water to ice is 80. The specific heat of ice at temperatures below 0oC is 0.5, which means that
to lower the temperature of ice by 1oC, 0.5 calories of heat needs to be removed. For all practical purposes, it is
assumed that fish have the same values for specific heat and latent heat as water. All this means that, if heat is
removed from fish at a constant rate, there will be a period during the fish freezing, where the temperature of fish will
not drop. This period lasts until approximately 75% of the water is frozen, when the temperature begins to drop again.
There are three stages to freezing fish fig.1). During stage 1, the temperature falls fairly rapidly to just below 0oC (and
it is called as initial falling rate period), during stage 2, the temperature remains fairly constant at about –1oC as the
bulk of the water (three quarters of the water) in the fish freezes (this stage is known as the ‘thermal arrest’ period)
and during stage 3 (final falling rate period), the temperature again drops and most of the remaining water becomes
frozen. The temperature at –1.1oC, freezing begins, and –5oC, about 80% of water is frozen, this range is termed at
‘Critical range’. The ideal thermal arrest is only 30min. Freezing is complete only when the equilibrium temperature
reaches –18oC(ie. the process should not be regarded as complete unless and until the product temperature has
reached –18oC at the thermal centre after thermal stabilization). It also shows that even at temperatures as low at –
30oC, a portion of water in the fish muscle still remains in the unfrozen state.
Under ideal conditions, fish should be frozen to –30oC in 2 hours.

Using simple mathematics, the amount of energy required to freeze fish can be calculated.
Suppose, if 1 kg fish at 25oC is to be frozen at –30oC. During stage 1, one calorie of energy has to
be extracted for each gram material for each drop of 1oC. In an example, the temperature of 1000g of fish will be
lowered from 25oC to –1oC, ie., by 26oC. The energy required will be equal to 1000 x 26 x the specific heat of water
(1) = 26000 calories or 26 kilocalories (kcal).
During stage 2, 80 calories of energy needs to be extracted for each gram material frozen. In the example, 1000g of
fish are to be frozen. The energy required will be equal to 1000 x the latent heat for freezing water (80), which equals
80,000 calories or 80kcal.
During stage3, 0.5 calories of energy needs to be extracted for each g material for each 1 oC drop in
temperature. In the example, the temperature of 1000g of fish will be lowered from –1 to –30oC, ie., by 29oC. The
energy required will be equal to 1000 x 29 x the specific heat of ice (0.5 = 14500 calories =14.5 kcal.
To summaries:
Stage 1, 1000 x 26 x 1 = 26 kcal
Stage 2, 1000 x 80 = 80 kcal
Stage 3, 1000 x 29 x 0.5 = 14.5 kcal
Adding these three figures gives 120.5 kcal, ie., to freeze 1kg of fish from 25oC to –30oC, 120.5 kcal of energy
is required.
From the above example, it is apparent that more than 50% of the energy extraction during freezing of fish
is taking place in stage 2. The thermal arrest period, where little or no drop in temperature is occurring, and this period
is a critical one, if a good frozen product is to be produced. Ideally, fish should pass through the thermal arrest period
as quickly as possible, due to the following reasons:

1. It is postulated that slow freezing produces large ice crystals in the cells of the fish, which can be larger than the
cells themselves and so break the cell walls. But it is not so, because it is evident that the walls of fish muscle cells
are sufficiently elastic to accommodate the larger ice crystals without excessive damage. Also, most of the water
in fish muscle is bound to the protein in the form of a gel and little fluid would be lost.

22
2. As the water begins to freeze in the flesh, the concentration of salts and chemicals in the remaining water rises.
This high concentration of salts and enzymes can cause accelerated autolysis and denaturation of protein
during slow freezing, This result in an inferior quality product and it is mainly due to denaturation of the protein.
Changes take place in some fractions of the protein as a result of freezing and since the proteins are altered from
their “native” state they may be said to be “denatured”, hence the term “protein denaturation”.
3. At temperature around 0oC, certain types of bacteria are still active and bacterial spoilage can still occur.
. Textural changes occur in fish, which have been frozen slowly, caused by the presence of large ice crystals and
denaturation of protein during the accelerated autolysis. In addition, a phenomenon known as ‘thaw drip’ occurs,
when slowly frozen fish are thawed. On thawing, the water which was originally bound within the cells are released
and considerable loss in weight can occur.
However, from a textural point of view, it is unlikely that a highly trained taste panel could detect the
difference between fish frozen in 1 hour and those frozen in 8 hours, Freezing times of 24 hours or more will almost
certainly result in an inferior product and very long freezing times can result in bacterial spoilage making the fish
unfit for consumption.

Chapter 2: Physical, chemical and thermodynamic properties of refrigerants

3.2.1. Refrigerant
It is any substance that absorb heat through expansion or vaporization and loses it threw condensation in a
refrigeration system. It is otherwise called as working fluid in a refrigeration system

Desirable properties of an ideal Refrigerant

1. Low boiling point

2. High critical temperature
3. High latent heat of vaporization
4. Low specific volume of vapour
5. Non-corrosive to metal
6. Non-flammable and non-explosive
7. Non – toxic
8. Low cost
9. Easy to liquefy at moderate pressure and temperature
10. Easy of locating leaks
11. Mixes well with oil
A refrigerant is chosen which has larger advantages and less disadvantages.
3.2.2. Thermodynamic properties
1. Boiling temperature
Refrigerant Boiling temperature (ºC)
Should be low at atmospheric pressure. If it is high, R-12 (CCl2F2) - 29
compressor should be operated at high vaccum, R-717 (NH4) -33.3
reduces capacity and increase the cost of the system.
R.744 (CO2) -73.6
2. Freezing temperature: R-764 (SO2) -10

Should be below the operating temperature of

Refrigerant Boiling temperature (ºC)
evaporator. Freezing temperature of most of
refrigerants are below -35ºC R-12 - 157.5
R-717 -35
3. Evaporator and condenser pressure R.744 - 56.7
R-764 - 75.6
Both the evaporating and condensing pressure should
be +ve (i.e. above atmospheric pressure). The + ve pressure are necessary to prevent leakage of air, moisture into
the refrigeration systems. It also permits easier detection of leaks.

23
Critical temperature & pressure:

It is the highest temperature at which refrigerant can be condensed to liquid, regardless of high pressure.

Refrigerant Critical temperature (ºC) Critical pressure (bar)

R-12 112 41.2
R-717 133 113.86
R.744 31 73.8
R-764 157 78.7

Coefficient of performance and power requirements

For an ideal refrigerant operating between -15ºC evaporator temperature and 300C condenser temperature. The
theoretical coefficient of performance for the reversed cycle is 5.74.

Refrigerant Coefficient of performance Power HP/TR

R-12 4.70 1.00
R-11 5.09 0.93
R-717 4.76 0.99
R.744 2.56 1.84
R-764 4.87 0.97
3.2.3. Latent heat of vaporization
A refrigerant should have a high latent heat of vapour at the evaporator temperature. It results in high
refrigerating effect per kg of refrigerant and which reduces the mass of refrigerant to be circulated per ton of
refrigerants.

Refrigerant Latent heat of vapour at 15ºC

(Kcal/kg)
R-12 (CCl2F2) 37.97
R-717 (NH4) 314.42
R.744 (CO2) 65.44
R-764 (SO2) 94.27

Specific volume It indicates the theorentional displacement of the compressor (i.e volume of suction vapour to
compressor).

3.2.4 Physical properties of refrigerants

Stability and inertness

An ideal refrigerant should not decompose at any temperature normally encountered in the refrigerating system.
This is due to the reaction with metal. To avoid this, a refrigerant should be inert with respect to all material used in
refrigerated system.

Corrosive property

Freon groups are non-corrosive with all metals. NH4 is used only with steel and Sn. S02 is non-corrosive to all metals
in the absence of water because it reacts with water and forms sulphuric acid.

24
Viscosity:

The refrigerant in the liquid and vapour states should have low viscosity because the heat transfer through
condenser and evaporator is improved at low viscosities.

Thermal conductivity

The refrigerant in the liquid and vapour states should have high thermal conductivity. This is required in finding the
heat transfer coefficient in evaporator and condensers.

Leakage tendency:

It should be low and the leakage of refrigerant should be easily detected by its pungent odour and also by using
burning sulphur candle which forms white fumes of ammonium sulphate in the presence of ammonia.

Cost

It is not so important for small refrigerant units but it is very much important in high capacity refrigerant system like
industry and commercial. Ammonia is the cheapest one. So it is widely used in large industrial plant such as cold
storage and ice plants. R-22 is costlier than R-12. The cost of loss due to leakage is also important.

3.2.5. Refrigerant

It is a substance in the vapour state which is used as a working fluid in refrigerators. In a refrigerating machine, heat
is transferred from cold body to hot body through refrigerants. It is also a cooling agent of refrigerating system. The
choice of refrigerant depends on ability to remove heat, toxicity, density, availability, cost etc.

Classification

1. Primary refrigerants
2. Secondary refrigerants

Primary refrigerants

These are the refrigerants which cool the substance or space directly by absorbing latent heat. It is also known as
direct expansion system Eg. Ammonia, Freon, SO2, CO2 etc.

Ammonia (NH3)

o Used for commercial purposes. Mainly in cold stored and ice plants.
o The boiling temperature of NH3 is -33oC
o It is colorless gas with a sharp pungent smell
o Has good thermodynamic properties
o It is neutral to all metals, highly soluble in oil.
o It’s solubility increases with increasing pressure and decreasing temperature
o Used free small and medium refrigerating capacities
o Volatile and non toxic but in higher concentration suffocation occur due to lack of O2

SULPHUR DIOXIDE (SO2)

o Previously used in household refrigerators

o Toxic, non-explosive and non-flammable, non-corrosive
o Irritant to human body
o Non mixable with oil
25
 o Has pungent odour and low latent heat value

CARBON DI OXIDE (CO2)

o Used as dry ice (solid from of CO2)

o Not used as refrigerant
o Colourless, odourless, non-toxic, non explosive, non-corrosive, non-flammable
o Operating pressure is very high
o Not mixable with oil
o As it is colourless and odourless gas, leaks cannot be detected easily
o Safest of all
o Its thermodynamic properties are not desirable for refrigeration except in cryogenic applications
o Two forms of CO2 used to freeze foods are spraying liquid CO2 & CO2 snow.

3.2.6 Freon group

o They are fluorocarbons of methane and ethane series.

o They contain 1 or more of these halogens (chlorine, bromine, fluorine)
o Non toxic, non-flammable, non-explosive, non- corrosive, non-irritant to human body and eyes.
o Odourless, colourless
o Will not react with food product stored in the refrigerated space.
o Will not react with lubricating oil.
o Has excellent thermodynamic properties
o Only disadvantage is ozone layer is damaged.

FREON 11 (Trichloro monofluro methane – CCl3F)

o Has the boiling point of 74.8ºF.

o Used in air conditioning plants in a larger level factories, theatres

FREON 12 (Dichloro-difluro methane –CCl2F2)

o Mostly used in domestic and commercial refrigeraters (in ice cream cabinets, display cabinets, deep
freezer)
o It is very widely used, colourless gas with mild odour
o Heavier than air
o Does not dissolve in water, moisture content should not exceed 0.0025% by weight
o It does not react with metals like ferrous, aluminium, phosphor bronze
o It attacks copper, copper alloys, zinc and bronze and dissolves in water
o It does not react with lubricating oils in the absence of moisture, but oxidizes them in the presence
of water vapour

FREON 13 (Mono chloro-trifluromethane) (CCl F3)

o Widely used
o Somewhat inferior in thermodynamic properties
o FREON 21 – (dichloro - monofluoroethane) (CHCl2F)
o FREON 22 – (monochloro – diflueroethane) (CHClF2)
o There are used in small refrigeration plants
o Used to obtain moderately low temperature (-50ºC)

26
 3.2.7 Secondary refrigerant

 Substances that take away heat from the medium to be cooled and give it to the boiling refrigerant are
called secondary refrigerants. They do not change their physical condition
 Cheap and harmless, chemically neutral to metals and packing material
 Should have a low freezing point and large heat capacity
 The refrigerants are brine which is used as intermediate fluid between evaporator and the substance or
space to be cooled. They cool the substance and the space by absorbing their sensible heat, which is also
being called having indirect expansion system.
 Eg. Brine solution made of calcium chloride or sodium chloride
 Choice of brine depends on temperature to which a material is to be cooled and industrial process in which
it is to be used
 For lower temperature (-18 to -23oC), CaCl2 is used, it is very costly
 NaCl is used when it is desired to have direct contact between brine and product.
 Freezing point of brine depends on its concentration
 Cheapest secondary refrigerants are water and air but their application is limited. Since, water has a high
freezing point (0oC) and air has a low heat capacity.

3.2.7 Secondary refrigerant

 Substances that take away heat from the medium to be cooled and give it to the boiling refrigerant are
called secondary refrigerants. They do not change their physical condition
 Cheap and harmless, chemically neutral to metals and packing material
 Should have a low freezing point and large heat capacity
 The refrigerants are brine which is used as intermediate fluid between evaporator and the substance or
space to be cooled. They cool the substance and the space by absorbing their sensible heat, which is also
being called having indirect expansion system.
 Eg. Brine solution made of calcium chloride or sodium chloride
 Choice of brine depends on temperature to which a material is to be cooled and industrial process in which
it is to be used
 For lower temperature (-18 to -23oC), CaCl2 is used, it is very costly
 NaCl is used when it is desired to have direct contact between brine and product.
 Freezing point of brine depends on its concentration
 Cheapest secondary refrigerants are water and air but their application is limited. Since, water has a high
freezing point (0oC) and air has a low heat capacity.

3.2.8. Physical characteristics of refrigerants

o Boiling and condensing temperature and pressures

o Freezing temperature
o Critical temperature
o Discharge temperature
o Latent heat of vapourisation
o Specific heat
o Density
o Viscosity

Boiling and condensing temperature and pressures

o The evaporator and condensing temperatures determine the pressures

o The maximum condensing temperature is largely affected by climatic condition
o It is desirable to select a refrigerant whose saturation pressure (at minimum operating temperature)
is a few pounds above atmospheric pressure.

27
Freezing temperature
Should have low freezing temperature to avoid operational obstruction by the refrigerant itself
Critical temperature
Should be well above the maximum condensing temperature

Discharge temperature
 High discharge temperatures from the compressor should be avoided
 It causes some refrigerant breakdowns as well as poor lubrication effectiveness

Latent heat of vapourization

 Heat which converts the refrigerant from the liquid state to vapour
 It should have a higher value

Specific heat : Higher specific heat is not desirable

Density : Low vapour density refrigerants are preferred

Viscosity : Low viscosity of the liquid refrigerant is desired to reduce the pressure drop in the lines

3.2.9. Chemical properties of refrigerants

o Toxicity
o Flammability and explosion hazard
o Refrigerant odours

Toxicity

o Rated based on its effect on human beings over specified periods

o Should be non toxic and non irritation

Flammability and explosion Hazard

o Should be non-flammable and non-explosive

Refrigerant Odours

o Can be both an asset and a hazard

o Makes it easy to detect the leaks but at the same time may contaminate foodstuffs in storage

Chemical properties

o Should be non-flammable
o Should be non-explosive
o Should be not-toxic
o Should not react with lubricating oil
o Should not react with moisture

Should not contaminate the food materials kept inside the refrigerating system

28
 3.2.10. Chemical properties

1. Flammability

It is an important characteristic in the selection of refrigerant. Ammonia and methyl chloride refrigerant will burn
but are explosive under unusual conditions only. CO2, N2, NO2 and fluorocarbon are nonflammable and non-
explosive.

2. Toxicity

All fluorocarbon refrigerant become toxic when mixed with certain percentage of air. Ammonia and SO2 are highly
toxic and not used in air conditioning. The use of toxic refrigerant is limited to cold storage.

3. Miscibility

The ability of refrigerant to mix with oil is called miscibility. The degree of miscibility depends upon the temperature
of oil and pressure of refrigerating vapour. The Freon group are highly miscible refrigerant while ammonia, CO2, SO2,
and methyl chloride are relatively non-miscible. The non – miscible refrigerant require larger heat transfer surface
due to poor heat conduction properties of oil. The miscible refrigerant give better lubrication.

4. Effect on perishable materials:

The refrigerant used in cold storage and in domestic refrigerators should be such that in case of leakage, it should
have no effect on the perishable material.

The Freon group have no effect on dairy products, meats, vegetable, flowers & fruits. Methyl chloride also have no
effect on materials. But SO2, ammonia affect the material.

NH4 easily dissolve in water and becomes alkaline in nature.

Since most fruits and vegetables are acidic in nature, therefore ammonia reacts with these products and spoils
the taste.

3.2.11. Economic and thermodynamic properties

 Should have high value of latent heat of vaporization

 Should have high thermal conductivity
 Should have high electrical resistance
 Should have low specific volume
 Should have low specific heat
 Should have low freezing point (than the maximum temperature of the refrigeration system)
 Should have low boiling temperature
 Should have low leakage tendency. Leakage should be detectable
 Above all the refrigerants should not be expensive

Chapter 3: Types of freezer

3.3.1. Introduction

The three basic methods of freezing fish are:

1. Blowing a continuous stream of cold air over the fish – air blast freezers
2. Direct contact between the fish and a refrigerated surface – contact or plate freezers

29
 Immersion in or spraying with a refrigerated liquid – immersion or spray freezers
3.3.1.1. Air blast freezers

The advantage of the blast freezer is its versatility. It can cope with a variety of irregularly shaped products and
whenever there is a wide range of shapes and sizes to be frozen, the blast freezer is the best choice. Before going
on to describe the various types of air blast freezer, it is necessary to deal with some of the basic principles of air
blast freezer design and operation.

Designing air blast freezers

The use of air to transfer heat from the product being frozen to the refrigeration system is probably the most
common method used in commercial refrigeration. The natural convection of the air alone would not give a good
heat transfer rate; therefore, forced convection by means of fans has to be introduced. To enable the product to be
frozen in a reasonable time the air flow rate should be fairly high. Also, in order to obtain uniform freezing rates
throughout the freezer, the air flow requires to be consistent over each fish or package.

Frost build-up on the cooler is also more prolific on the upstream coils; therefore a cooler with a large frontal area
will be able to operate longer before a defrost is necessary.

Continuous air blast freezers

The belt has to be flexible, easily cleaned, non corroding, suitable for use in direct contact with food and should not
interfere unduly with either the freezing time or adversely affect product quality. Stainless steel mesh link belts or
chain link belts are mainly used for this purpose but they have certain disadvantages. Apart from being expensive,
they affect the appearance of the product. If fish are loaded directly on the belt, the crinkled or indented appearance
of the frozen product is not always acceptable. Open mesh belts can also give rise to difficulty when removing the
product after freezing and some weight loss may be incurred due to slight physical damage. Skin-on fillets can
usually be removed quite easily but skinless fillets and fish portions can stick to the belt and cause unacceptable
weight losses.

Continuous belt freezers can be constructed with either cross-flow or series-flow air circulation. In the series-flow
arrangement, the direction of air flow must be such that the coldest fish meet the coldest air. The design of the belt
entry and exit must keep the rate of air
infiltration to a minimum.

Fluidized and semi-fluidized freezers

One type of air blast freezer fluidizes the product
with a strong blast of air from below. The product
then behaves like a fluid and when poured into the
trough at the input, it moves along the length of
the freezer without mechanical assistance and
over-flows at the output. This type of freezer has
not had a wide application for fish or fishery
products. Small cooked and shelled shrimp is one
of the few fish products that have been
successfully frozen by this method.
3.3.1.2. Plate freezers

Plate freezers can be arranged with the plates horizontal to form a series of shelves and, as the arrangement
suggests, they are called horizontal plate freezers (HPF). When the plates are arranged in a vertical plane they form a
series of bins and in this form they are called vertical plate freezers (VPF). Modern plate freezers have their plates
constructed from extruded sections of aluminium alloy arranged in such a manner as to allow the refrigerant to flow
through the plate and thus provide heat transfer surfaces on both sides. Plate freezers are fitted with hydraulic
systems which move the plates together and apart.
30
Horizontal plate freezers

At present in India all the processing plants have HPF. Capacity of this freezer varies between 500kg to 1ton/load.
Each plates are connected with pressure resistant hose on both ends, i.e. suction line and discharge line. A hot gas
defrost arrangement is the quickest method to defrost an HPF, but even with this method, it may take 30 min or
more. The defrosted plates must be completely free from frost or ice and dried before the freezer is used again.

Vertical plate freezers

The main advantage of this type of freezer is that fish can be frozen in bulk without the requirement to package or
arrange on trays. This type of freezer is therefore particularly suitable for bulk freezing and it has also been
extensively used for freezing whole fish at sea. The maximum size of block made by this method is usually 1070 nm x
535 mm. Other dimensions however, can be produced in which the thickness can vary from 25 to 130 mm, and it will
depend on the fish to be frozen. The maximum weight and dimensions are also limited by the physical effort
required from the operator to lift the block, and by the ease with which it can be handled so that damage to the fish
is kept to a minimum.

In most cases, fish can be loaded between the plates without wrappers and water need not be added either to

strengthen the frozen block or improve the contact with the plates. Fish such as cod and haddock produce compact
blocks with a block density of approximately 800 kg/m3. It has certain operational disadvantages.

Defrost is essential to prevent fish sticking to the plates which are at a temperature below 0oC, and thus failing to
form a compact block. Freezing times are longer due to the poor contact being made with the plates and because of
the lower block density, more storage space is required for a given quantity of fish
3.3.1.3. Liquid nitrogen freezer

In this freezer, the product is brought into direct contact with the refrigerant. The fish on the stainless steel conveyor
belt initially come into contact with the counter current flow of nitrogen gas at a temperature of about –50oC. As the
fish progress through the pre-cooling stage of the freezer, the gaseous nitrogen partially freezes the fish and up to
50 percent of the product heat is extracted. The product then passes below the liquid spray where freezing is
completed by the boiling liquid. The last stage in the freezer provides a few minutes for the fish temperature to
reach equilibrium before the fish are discharged.

The main advantage of the liquid nitrogen freezer LNF is that freezing is very quick and the physical size of the
freezer is correspondingly small. The freezer is operated without the need for compressors, condensers or coolers;
therefore maintenance requirements are minimal and the power required to operate the freezer is very low.
Liquid nitrogen must be retained in a vacuum insulated pressure vessel with continuous venting to keep the contents
cool and the internal pressure down. It is found that 0.5 percent of the stored contents is lost each day by this
method. In addition, about 10 percent has been estimated to be lost during the transfer of liquid from the tanker to
the storage vessel. This method of freezing is more expensive than most others. It is four times more costly than
conventional air blast freezing. Although the freezer is small and there is no refrigeration machinery requirement,
storage space and access is required for the liquid nitrogen tank. The main disadvantage of this type of freezer in
most developing countries like India is that delivery of nitrogen could be expensive and there may be no guarantee
of regular supplies.

3.3.1.4. Carbon dioxide freezer

This type of freezer has been known for a long time and uses liquefied carbon dioxide which is usually a by-product
of another industrial process.

The liquefied carbon dioxide is injected into the freezer has direct contact with the product. In this respect, it is
similar in operation to an LNF. With large units, it is economically feasible to recover the carbon dioxide and about

31
80 percent of the refrigerant used can be reliquefied. Carbon dioxide can be contained in insulated vessels at a
moderate pressure and losses during storage are therefore negligible.

High levels of carbon dioxide in the factory air are dangerous, therefore a freezer using this refrigerant must be
vented and the gas discharged outside the building. Again, as in the case with other types of freezer which rely on
regular supplies of refrigerant, carbon dioxide freezers would not be suitable for use in remote areas.

3.3.1.5. Immersion freezers

In this method a liquid refrigerant is used for the removal of heat from a product. Favourable freezing rates can be
achieved. Liquid can remove more heat per unit volume than gas (eg. air) but, like gas, a stagnant boundary layer is
form which slows the transfer of the heat. Liquids used for heat transfer must therefore be circulate over the
product. Difficulties due to high viscosity often arise when a low temperature liquid is used.

Chapter 4: Methods of protective treatments

3.4.1. Brining
To minimize drip loss, fillets/steaks of fish are immersed in brine for a short time and this process is known as firming
of fillets/steaks. There are several advantages in this type of firming.

1. When fish fillets are dipped in strong brine, the proteins in the outer surface of the fillet gets coagulated due
to which firming takes place.
2. The layer of salted out proteins insulates the muscle tissues from the negative influence of outside factors.
3. It favours the retention and preservation of natural extractives and flavour compounds.
4. It minimized drip loss.

Effect of salt concentration in brine and brining time

The treatment of fish fillets with brine is a precision operation. When the salt concentration is less than 5% proteins
get solubilized and lost in salt solution. If it is more than 15%, tissue proteins coagulate due to “salting out” of the
proteins and the texture becomes tough. If the duration of dip treatment is more, the product becomes salty.
Therefore, brining treatment should be done in right concentration for a brief period of time.

Method of brining: Before the treatment with brine, the fillets are washed properly, because brine treatment on a
clean surface is more effective. Salt concentration in brine should be 10-12% (sp. gr. 1.07 to 1.09). Brining temperature
should be less than 15OC. In dip treatment the brining time is 1 to 2 min. However, there is scope to change the salt
concentration and duration of time for treatment depending on species of fish. The brine should be changed after
every dip treatment. Addition of antimicrobials such as sodium benzoate, benzoic acid, parabens, sodium nitrite and
antioxidants like ascorbic acid can be added to brine in order to maximize its effectiveness. However in fatty fish
bringing promotes fat oxidation in it and hence the results are not encouraging.

3.4.2. Advantages of brine treatment

1. Treatment with brine destroys the microorganisms present on the fish fillet to a great extent. However, salt
should not contain Ca++ and Mg++ impurities which are responsible for salt odour and bitter taste.
2. Prevention of belly bursting by brine treatment: During chilled storage and during freezing and thawing, belly
bursting is observed in sardines (poor appearance, viscera comes out) which is a major problem in small fish
such as sardine. If sardine is dipped in 15% brine for 30 min and then frozen, the problem of belly bursting can
be minimized. By this method, due to coagulation of proteins the belly wall becomes tough. However the
treatment promotes oxidation of fat and hence the shelf life in frozen storage gets reduced.

32
Phosphate treatment : PolyPO4 treatment minimizes drip loss in frozen fish and fishery products. PolyPO 4
(Polyphosphate) treatment may be given separately or along with brine treatment. In cod fillets drip treatment in
0.0844 < (4%) polyPO4 solution minimized significant amount of drip loss.
Role of Polyphosphate in minimizing drip loss :
In frozen fish, water holding capacity of tissues comes down due aggregation and protein denaturation and hence
results in increased drip loss. The important reasons for drip loss are given below.

1. Freezing changes ionic environment of fish muscle.

2. Reduction in pH due to production of lactic acid.
3. Degradation of nucleotide which contains active phosphorus.
4. Formation of actomyosin from actin and myosin.

The role of polyphosphate in minimizing driploss and preservation of quality of fish is explained as follows.

1. Balanced action of tripoly PO4, organic phosphate such as ATP and other phosphates leads to break down of
the complex protein actomyosin to constituent proteins actin and myosin resulting in increased water holding
capacity of muscle.
2. Cell fluids of fish (salted) dipped in brine require -22OC for complete freezing due to increase in salt
concentration. Therefore liquid present in such cell fluids remain unfrozen and atmospheric oxygen dissolve
in such a liquid and penetrates inside the tissues. This accelerates oxidation process. However, if 12%
tripolyPO4 is mixed with brine, the freezing point of tissue fluids will only be -3OC. Therefore proportion of
unfrozen liquid will be greatly reduced and also concomitant oxidation. This is very much useful in fatty fishes.
3. Polyphosphate acts as a chelating agent and chelate proxidant metal ions, thereby minimizing catatytic action
of metal ions in fat oxidation.

3.4.3. Use of Polyphosphate in freezing of fish

Dip treatment in phosphate solution prior to freezing gives better results. In prawns, in addition to dip treatment in
phosphate solution, it may be mixed with glaze water.
Quantity of polyPO4 to be used
Fairly high proportion of phosphate ion is required for breakdown of complex acto-l-myosin into its constituent
proteins. Among various phosphates, tripolyPO4 and pyropolyPO4 gives the best results.
Use of tripolyphosphate solution of 0.16 M ionic strength (8%) and 4.5% pyropolyPO4 along with salt to increase the
ionic concentration to 0.4 M and 0.35 M respectively increases the water holding capacity of muscle tissues effectively.
Among the above, tripoly phosphate is extensively used but its effect depends on the species of fish. Though tripolyPO4
solution and brine can be used separately, fish and fillets are treated with pyropolyPO4 and brine together very good
effect was observed.
Polyphosphate should be used in such a way that it is absorbed quickly into fish muscle. It has been found that the
enzymes and digestive juices convert the polyPO4 in to orthoPO4 in the body and hence it does not pose any health
hazard. However it is better to take proper care while using depending on its desired effect on the product.
Antioxidant treatment
Rancidity is the commonly observed storage problem in fatty fish. To minimize the problem of rancidity, antioxidants
which are commonly used in edible oils and fats have to be used in fish. Rancidity in oils and fats is controlled by
following methods.

1. Removal of pro-oxidant chemical compounds or inactivation of these factors.

2. Packaging of fish so that oxygen is prevented from coming into contact with fish surface and subsequent
penetration inside the tissues.
3. Before or after freezing fish may be treated with antioxidants. Enzymes responsible for fat oxidation may be
inhibited.

Phenolic antioxidants, even though very effective, they are not soluble in water. Some of the methods used for
application of antioxidants for frozen fish and fillets are as follows.

33
 1. Water soluble antioxidants such as ascorbic acid penetrates the tissues of fish through tissue fluids and helps
in arresting oxidation process. It is effective against salmon, trout, herring and tuna during frozen storage. It
is also effective against colour change in pomfrets. Ascorbic acid also acts as a synergist when used along with
BHA, tocopherol and hydroquinone. Ascorbic acid in glaze increases the shelf life of mackerel and pomfret
during chilled and frozen storage.
2. Glaze containing ascorbic acid and citric acid mixture is effective against fatty fish and onset of rancidity can
be delayed for a long time.

Oil soluble antioxidants are useful in minimizing rancidity in fatty fishes. This is because the oil of fish is on the
surface and the antioxidants dissolve quite easily into such oil and penetrates into the tissues. The antioxidant power
of BHA and Ascorbic acid double when used as a mixture.
3.4.4. Methods of using antioxidants

1. Dip treatment prior to freezing

2. In glaze water

Water soluble antioxidants can be easily dissolved in water and can be directly used. However, oil soluble antioxidants
are first emulsified in small quantity of oil in water type and used. When gallates are used there should be no contact
with Fe++ and pro oxidant metal ions may be bound by chelating agents and make them unavailable in oxidation
reactions. Nitric acid, phosphoric acid, sodium phosphate citrates, tartarates and ethylene diamine tetra acetic acid
(EDTA) are some of the important chelating agents. These may be used separately or along with anti oxidants.
Commonly used antioxidants
Antioxidants are of 2 types 1) Water soluble and 2) Oil soluble.
Antioxidants used in fish processing are as follows
Compound Nature Max. Percentage used
1) BHA Oil 0.02
2) Salts of gallic acid
a) Propyl gallate Water 0.01
b) Octyl gallate Oil 0.01
c)Do decyl gallute Oil 0.01
3) Nor-dihydrogui acetic acid Oil 0.01
4) Ascorbic acid Water 0.25-2
5) Sodium ascorbate Water 0.25-2
6) Sodium sulphide Water 0.25-2
7) Tocopherols Oil 0.03
Amino acids (cystine, Water 0.03
8)
histidine, tryptophan)
9) Synergist (citric acid,sorbitol) Water 0.03

As antioxidants break the chain of free radical formation, oxidation is prevented. However, this is easy in the case of
almost pure oils but in the case of fish it is quite difficult because the action of antioxidant depend on several factors
such as

1. Property of oil in the tissues.

2. Penetration of antioxidants into tissues .
3. Presence of Pro-oxidant materials.

Therefore some antioxidants are effective only against certain species of fish. Some problems associated with the use
of antioxidants in fish are

1. Determination of an antioxidant for a particular species of fish.

34
 2. Medium and method of application of the antioxidant.

Chapter 5: Calculation of freezer refrigeration load

3.5.1. Calculation of freezer refrigeration load

The individual items to be taken into account in a refrigeration load calculation depend on the type of freezer. It
would be impossible to include all the eventualities in one sample calculation; therefore, a relatively simple one is
given below for a HPF and some notes have been added to help with other freezer calculations.

Load calculation

I. Number of freezer

20 t/day = 20,000 kg/day

20,000 ¸ 10 = 2,000 blocks/day

24 ¸ 2 = 12 cycles/day

2,000 ¸ 12 = 167 blocks/cycle

II. Fish load

20,000 ¸ 24 = 833 kg/h

Enthalpy at –30oC = 4.6 kcal/kg

Change in enthalpy = 81.3 kcal/kg

An approximate figure can also be calculated by using the following values:

a) Specific heat of fish above freezing, 0.9 kcal/kgoC

b) Latent heat of the fish, 60 kcal/k

c) Specific heat of fish below 0oC, 0.4 kcal/kgoC

Using these values, the above calculation for fish refrigeration load would be:

Heat to remove on cooling to 0oC

833 x 0.9 x 10 = 7497 kcal/h

Latent heat to remove 833 x 60 = 49980 kcal/h

Heat to remove on cooling to –30oC

833 x 0.4 x 30 = 9996 kcal/h

Total heat to remove from fish = 67473kcal/h

Total refrigeration requirement with allowances:

35
Add 30% = 142681 kcal/h

The calculation of fish load gives refrigeration requirement to freeze the fish only. Depending on the type of freezer
used, other heat loads such as fan heat, pump heat from circulating pump, heat leak through freezer insulation, heat
load due to pallets, trays, trolleys, defrost procedure, air infiltration,internal lighting etc. have to be taken into
account and added to this value to determine the total refrigeration requirement.
3.5.1.1. Freezing time

Following are the Variables which affect freezing time

1. Freezer type
2. Freezer operating temperature
3. Refrigeration system and operating condition
4. Air speed in an air blast freezer
5. Product temperature
6. Product thickness
7. Product shape
8. Product contact area and density
9. Product packaging
10. Species of fish

Freezer type: The type of freezer will greatly influence the freezing time. For example, due to improved surface heat
transfer, a product will normally freeze faster in an immersion freezer than in an air blast freezer operating at the
same temperature.

Operating temperature: The colder the freezer, the faster the fish will freeze.

Air speed in blast freezers: Freezing time is reduced as the air speed increased.

Product before freezing: The warmer the product, the longer it will take to freeze. Fish should therefore be kept
chilled (+4oC) before freezing both to maintain quality and reduce freezing time and refrigeration requirement.

Product thickness: The thicker the product, the longer is the freezing time.

Product shape: The shape of a fish or package can have a considerable effect on its freezing time and this is
dependent on the ratio of surface area to volume.

Product contact area and density In a plate freezer, poor contact between product and plate results in increased
freezing time.

Product packaging: The method of wrapping and the type and thickness of the wrapping material can greatly
influence the freezing time of a product. Air trapped between wrapper and product has a greater influence on the
freezing time than the resistance of the wrapping material itself.

Species of fish The higher the oil content of the fish the lower is the water content. Most of the heat extracted
during freezing is to change the water to ice; therefore, if there is less water, then less heat will require to be
extracted to freeze the fish. Correspondingly freezing time will be less.
3.5.1.2. Calculation of freezing times

The more general from of Plank’s equation for calculation freezing time is:

Freezing time = L / V ( PD/D + RD2/fk )

36
Where

L = Heat to be extracted between the initial freezing point and final temperature (kcal/kg).

V = Specific volume of fish (m3/kg)

D = Temperature difference between the initial freezing point of the

Values for shape constants P and R
fish and the refrigerating medium (oC)

D = Thickness of product in direction or prevailing heat transfer (m) Shape P R

f = Surface coefficient of heat transfer (including effect of Sphere 0.167 0.042

packaging) (kcal/h.m oC)

Infinite 0.167 0.042
Cylinder
k = Thermal conductivity of frozen fish (kcal/h moC)

P and R= Constants which depend on shape Infinite Slab 0.500 0.250

Calculated freezing time – Example 1: What is the freezing time if all other conditions remain the same but the
operating temperature is –25oC.

Fish freezers at about –1oC, therefore in the measured freezing time, the effective temperature difference is 34ºC
(the difference between –35oC and –1oC). The effective temperature difference for the freezing time required is 24ºC
(the difference between 25oC and –1oC). Freezing time is inversely proportional to the temperature difference,
therefore the freezing time with an operating temperature of –25oC will be longer than for a temperature of –35oC
and can be calculated as follows:

200 x 34 % 24 = 283 min or 4h 43min

Calculated freezing time – Example 2 What is the approximate freezing time if all other conditions remain the same
and the block thickness is reduced to 75mm?

Freezing time is directly proportional to the square of the thickness since in this case the surface heat transfer
coefficient is high and the factor relating to the thickness of the block, PD/f, will be small. The new freezing time will
therefore be calculated as follows:

200 x 752 ¸ 1002 = 200 x 5625 ¸ 10000 = 112 min (1h 52 min)

3.5.1.3. Coding, Packing and Storage

A code slip bearing the marking of the numerical code of the processor, abbreviation of the name and type of
products, year, month and date of production shall be embedded on the frozen block. In the case of individually
Quick frozen (IQF) packs the code slips shall be placed in the primary containers.

An illustration of the coding is given below:

520 FSPD

11A10

Where in the above illustration

37
520 – the numerical code of the processor/ exporter

FS – Name of the product (Frozen shrimps)

PD – Type of the product (Peeled and Deveined)

11 – Year of processing (here 2011)

A – Month or processing (here January)

10 – Date of processing (here tenth day of the month)

The following abbreviations shall be used for the months of the year:

January - A

February - B

March - C

April - D

May - E

June - F

July - G

August - H

September - J

October - K

November - L

December - M

The frozen products shall be packed in primary container other wise known as wax coated board or duplex canton
and such primary containers shall be packed in seaworthy fibre board cardboard cartons of 5 ply minimum or any
other better type of cartons. In the case of Individually Quick Frozen products the pieces shall be arranged/wrapped
in moisture proof film. As per the specific requirements of the buyer the packing of frozen fish and Fishery products
without primary container/Duplex cartons shall be permitted. In such cases the products shall be placed in a master
poly bag and placed inside the master carton. The frozen material shall be stored in a room maintained at or below a
temperature of minus 18o C.

Chapter 6: Freeze drying fish

3.6.1. Introduction
Heat is used to dry foods or concentrates liquids by boiling to remove water and therefore preserves the food by
reduction in water activity. However, heat causes a loss of sensory and nutritional qualities. In freeze drying a similar
preservative effect is achieved by reduction in water activity without heating the food and sensory and nutritional
qualities are consequently better retained. However both, operations (freezing & drying) are slower than conventional
dehydration. Freeze drying has become an important commercial operation to dry expensive foods which have
delicate aroma or texture (for example instant coffee, mushrooms, fruit juices and sea foods).
38
 3.6.2. Theory of freeze drying
The boiling point of water depends on the overhead atmospheric pressure. At normal atmospheric pressure of 1 bar
(i.e., 760 mg of Hg) it boils at 100ºC. However, if it is held in a sealed container and overhead atmospheric pressure is
lowered than one by using vacuum pumps, then it boils at less than 100ºC. It is observed that the lower the pressure
(or the better the vacuum) the lower is the boiling point of water but the energy needed for evaporation of the same
quantity of water is more. The pressure, the corresponding boiling point of water and latent heat of evaporation are
presented in the following table.
Pressure (bar) Boiling Point of Water Latent heat of
O
C vaporization (kj/kg) of
water
1 100 2257
0.5 81 2305
0.2 60 2358
0.1 46 2392
0.01 7 2485

At OOC water exists an ice (solid phase) and if the overhead pressure is sufficiently low, solid ice will directly
sublime as water vapour without going into liquid phase. To understand the phenomenon more clearly it is essential
to refer to the phase diagram of water.

Pressure units conversion table :

Torr mm Hg Pascal
1 1 133.3
10-1 0.1 13.33
10-2 0.01 1.333

From the phase diagram it is clear that if the temperature of water and pressure is below the triple point it
exists as ice and it will directly pass into vapour phase.

If the overhead pressure of a food is held below 4.58 Tor (610.5 Pa) and the food is frozen, when the frozen food is
heated solid ice sublimes directly to vapour phase without melting. Water vapour is continuously removed from the
food by keeping the pressure in the freeze drier cabinet below the vapour pressure at the surface of ice by removing
the water vapour with a vacuum pump and condensing it on refrigeration coils or plate or drum. As the drying
proceeds, the sublimation process continues as long as latent heat of sublimation is provided/supplied. The latent
heat of sublimation is either conducted through the food to the sublimation front or produced in the food by
microwaves. The pressure, sublimation temperature in the frozen state and corresponding latent heat of sublimation
are presented below.

In freeze drying, foods are dried in two stages, first by sublimation to approximately 15% moisture content (on wet
weight basis) and then by evaporative drying of unfrozen water to 2% of moisture content (W.W.B). Desorption is
achieved by raising the temperature in the drier to near ambient temperature while retaining the low pressure.
The rate of drying depends mostly on the resistance of the food to heat transfer and on the resistance to vapour
flow.
3.6.3. Principle of the equipment

Effectiveness of a freezes drier depends on rate of heat transfer to the sublimation front and rate of mass
transfer.There are three methods of heat transfer.

Heat transfer through the frozen layer. The rate of heat transfer depends on the thickness and thermal conductivity
of frozen layer. As the drying proceeds, the thickness of ice is reduced and the rate of heat transfer increases.
39
Heat transfer through dried layer. The rate of heat transfer to sublimation front depends on the thickness of the dried
layer and the surface area of food, thermal conductivity of dry layer and the temperature difference between the
surface of the food and ice front. The dried layer has very low thermal conductivity and therefore offers high resistance
to heat flow. As drying proceeds, this layer becomes thicker and the resistance increases.
Heating by microwaves : Heat is generated at the ice front and the rate of heat transfer is not influenced by thermal
conductivity of ice or dry matter or the thickness of dry layer. However it is difficulty to control microwave heating.

Rate of mass transfer: It depends on the water vapour pressure gradient inside the drying chamber which is
controlled by

1. The pressure in drying chamber

2. Temperature of the vapour condenser – both should be as low as possible.
3. Temperature of ice at the sublimation front and it should be on high as possible (at OoC) without melting.

Equipment : freeze drier consists of a vacuum chamber containing trays to hold food during drying, source of heat to
supply latent heat of sublimation, refrigeration coils to condense vapour with automatic devices to keep maximum
area of the coil free of ice for vapour condensations and vacuum pump to remove non expensible vapour.
Types of equipments :

1. Accelerated freeze driers : Food is held between expanded metal mesh for heat transfer.
2. Radiation freeze driers : In paired radiations from radiant are used. Heating is more uniform. Close contact
between food and the heat is not necessary; float trays can be used and cheaper. Easier to clean.
3. Secondary drying to remove bound water molecules from the primary dried materials.

1) Freezing : Freezing the product temperature is reduced to -50OC at one atmospheric pressure to ensure that it is
below the eutectic point.
2) Primary drying : This is a dynamic process involving transits of ice phase to vapour phase directly. This leaves the
solid matrix formed in the freezing phase but without the ice crystals. During drying, an interface is set up which comes
from the exposed surface to vacuum system to the surface where heat source is in contact or moves deeper in radiant
heat. At this interface, actual drying takes place. There are number of factors which will indicate the completions of
primary drying process.
1) Pressure stability : During drying, pressure changes slowly until it stabilizes at an ultimate low level. The process is
considered as complete where the constant pressure is maintained for 1 to 2 hours.
2) Product temperature rise. When heat energy is not required for sublimation, the overall product temperature will
rise until equal or close to the chamber temperature. This can be detected with a temperature probe and displayed.
3) Pressure rise test. If an isolation valve is available, it will be possible to isolate the drying chamber. If the material is
not dry, then the pressure will rise quickly (due to vapour pressure of the product).
Secondary Drying: This is part of the process where remaining small proportion of “locked in” water is removed (
Usually 2.5% by volume). This is not always considered necessary and depends on the product.The locked in or
bound water is removed form the products by supplying additional heat energy to the system. This is achieved
either by increasing the temperature or by improving the thermal contact between the product and the existing
of heat source.

4. Effect of freeze drying on food : Freeze dried foods have high retention of sensory and nutritional qualities
and shelf life of more than 12 months when correctly packed. Aroma retention is 80-100%. Shrinkage is very
little and texture is well maintained. Allows rapid and full rehydration but the product is fragile and require
protection from mechanical pressure. However, the open porous structure of the food may allow oxygen to
enter and cause oxidative deterioration of lipids and subsequent browning. Changes in thiamine and ascorbic
acid content during freeze drying are moderate and there are negligible losses of other vitamins. Therefore
vacuum or inert gas packaging is required. Microwave and dielectric freeze driers

40
 3.6.4. Freeze drying process
Freeze drying cycle consists of 3 stages.

1. Freezing.
2. Primary drying to remove water ice.

Unit 4: Thawing of fish

Chapter 1: Methods of fish thawing
4.1.1 Introduction
Thawing is a process whereby heat is introduced into the frozen product. In order to raise its temperature above
freezing point, the rate of heat input required to thaw 1 ton of fish per hour is approximately 85 KW. Thawers or
methods of thawing can be divided into two broad classes.
1) Those which depend only on the conduction of heat through the thawing material.
E.g. Thawing in air, thawing in water and thawing between heated plates.
2) Those which do not depend on the conduction of heat through the thawing material.
E.g. Dielectric thawing and electric resistance thawing.
There are also other methods such at infra red and microwave heating, but they are not of interest for thawing on a
large scale
4.1.1.1 Thawing in air

a) Natural convection:

It is sometime convenient to thaw at room condition, if conditions are suitable. Temperatures above 18ºC should
not be used. Lot of space and labour is required for thawing on a large scale by this method and the introduction of
humidification to prevent slight drying of the fish is difficult.
b) Air blast:
In this method, a current of air is directed on the frozen block and the increased air velocity reduces thawing
time. The rate of thawing is mainly dependent on the resistances of the fish itself to the flow of heat. At the air
velocity of 6m/sec, there is very little resistance, which is optimum. In modern air-blast thawers with the
recirculation of humid air at 20oC( the maximum, temperature that should be employed), the 10cm block of frozen
fish requires about 5 hours for thawing before they can be filleted. The fish can be separated easily after about 3
hours, which reduces the thawing

i) Continuous:
Continuous air-blast thawers have been built in two designs. Cross flow and parallel flow. In the cross flow
design, the direction of air flow is across the conveyor at right angle. Typically there are five conveyors, one above the
other. The fish travels horizontally along the top conveyor until when it reaches the end, it falls to the conveyor below
which is traveling in the opposite direction and so on. In order to achieve high velocity, capacity of the fan should be
high with this arrangement and use of fan power is more than 30 KW for a unit fish. Thus a large portion of the heat
for thawing is obtained from the fan.
In the parallel flow thawers, the air travels in the same direction as the conveyor. With humid air, most of the
heat transfer to the fish takes place by condensation at water vapour and an air temperature drop at 300C can be
tolerated. The fan capacity is relatively low and therefore, the fan power is about 7.5 KW for the 1 ton/hour unit and
the face area of the heater and humidifier in conveyor is, in a single line occupying a greater floor area, than a cross
flow thawers. In the other, the conveyor is divided into three sections, one above the other.

ii) Batch:
The largest model of batch air-blast thawers has five compartments each holding 1 ton of 10cm blocks of
whole fish on two trolleys with suitably- spaced shelves, giving an output at about 1 ton/hour. Air flow is in series
through the five compartments in a line. Periodic reversal at air flow can be employed to even out differences
between compartments. The thawers can also be used on a semi continuous basis by inserting and withdrawing
trolleys at intervals of 1 hours. More labour for loading and unloading is required than for continuous thawers, but
41
the batch thawer is simple and compact. The 20 KW fan heaters and humidifier are located above the fish
compartments. Batch thawers with small fans giving moderate velocities, in which blocks of fish are placed on racks
or shelves, have been used for thawing frozen fish for over periods of up to 10 hours.
4.1.1.2 Water thawing

a) Immersion:

The thawing of blocks of whole in water has been shown to give good results, but has been used only to a
limited extent for thawing on a large scale. The thawed product can be as high in overall quality and suitable for
processing. Pump capacities for water thawers need not be great, because the water velocity required for good heat
transfer is 0.5 cm/sec, giving thawing rates equal to those for air-blast. It is probably necessary, to use somewhat
higher velocities, perhaps 2cm/sec or more, in order to give a good degree of mixing of the water, preventing
stratification and to provide reasonably quick response to the heater, particularly for warm up with initially cold
water. Too high velocities induce turbulence, causing foaming at the surface of the water and possibly lead to water
logging. The temperature of the water should conserve heat, it is better to change the water once or twice in 24 hour
so as to limit the degree of contamination of the water.
b) Spray:
In this, larger thawers are used, in which heated water is sprayed directly on the fish. The blocks of fish travel on a
conveyor, which is made up of a series of open mesh, wire baskets in a housing ventilated by a small fan. The water
is recirculated through a special filter and cleaner section. The thawing time is claimed to be at least as short as in
air, blast thawing. Hygienic condition is maintained in the thawers because of the working action of the sprays of
clean water. It takes about 5 hours for the 10cm block of whole fish to be thawed completely, in a thawer with are
output of 1ton/hour.
4.1.1.3 Thawing between heated plates

This type of thawer is similar to a plate freezer except that warm fluid, normally at 20ºC is circulated throughout the
(metal) plate. The product is held in contact with the plates throughout the thawing period which is about 8 hours
for complete thawing of the 10cm block of whole fish. Thawing is more even than on air blast thawing, with the
result that the fish can be filleted sooner, typically 7 hours after an initial period at 4 hours in the thawer as opposed
to 12 hours in air-blast thawing. Little power is required excluding the heat for thawing, the plant is compact and
robust

a) Dielectric thawing:

In this method, the frozen material is placed in a high frequency electrical field between two flat metal plates. An
alternating voltage at 5,000 V and 40MHZ is applied to the plates and hence heat is produced in the electrically
charged particles in the food. The plates are fed from a generator and suitably shielded to confirm the field. Unless
the food is homogenous and regular in shape, there is possibility for uneven heating so that the parts of the material
are cooked while other parts remain frozen. This can be minimized in block at fish continuously conveying them
through the field and restricting the power density according to the degree of thawing. Thus, a typical industrial
thawer for blocks at fish includes a conveyor belt and six generators each with a rating of 20KW giving a nominal
output of 1 ton /hr with 10 cm blocks of whole fish. Thawing is very rapid so that the total length of conveyor has
been much shorter than that for continuous air-blast thawers. The thawing time for blocks of large white fish is
about 80 min, and for blocks of small fatty fish is about 20min.

b) Dielectric and water:

The dielectric thawer has a thermal efficiency of about 70%. Cooling water for the generators absorbs the waste
heat. It has been suggested that heated water be used for the thawing either before or after the dielectric thawer, a
hybrid system, in order to increase the output and thermal efficiency and reduce the risk of overheating. Since the
heat conduction is rapid during the early stages, the 10 cm block of whole fish can be 30% thawed in less than 30min
in water at 15oC but the complete thawing takes up to 80 min at the temperature at 15oC.

c) Electric resistance thawing:

42
In this method, fish can be heated by placing it between two electrodes used applying a low voltage so that a current
flows through. Like dielectric heating, it is difficult to achieve even heating unless the item is homogenous and
regular in shape. It is convenient to partially heat the fish by a heat conduction method such as immersion in
circulating tap water and thawing by heat conduction is rapid during the early stage. Usually the time taken for
water immersion and electric heating should be roughly equal. The total thawing time is between 20 and 60 min
depending on the type at blocks, at least twice as fast as thawing in air or water. A complete batch thawer of this
type has plates and blocks of fish arranged in layers one above the other in the same way as a horizontal plate
freezer. The plates are heated electrically or by passing worm fluid through them during the early stage at thawing
when heat conduction is employed and act as electrodes during the stage of electric resistance thawing.

d) Vacuum thawing

Vacuum thawers consist of airtight chambers in to which the fish is loaded using trolleys. The temperature of the
water vapour is typically between 18oC and 20oc and the vapour condenses on the cold surface of the fish where the
latent heat of vaporization is absorbed by the fish meat transfer to the product is much more rapid than the previous
method of thawing. Equivalent thawing fishes may be reduced from 20hr in air to less than 1 hr in a vacuum thawing
unit and the water usage at such units is low. However, care must be taken that gases released from the fish as
thawing proceed do not cause rupturing at flesh.

The advantages of this method of defrosting are that with the exception at a vacuum pump there are no moving
parts, hence the possibility of mechanical breakdown is reduced compared to the previous methods. Also the vapour
is distilled from water in the tank it condenses on to the fish in a pure state, so there is no risk of contamination from
bacteria which may accumulate within the system.
4.1.1.4 Choice of method
Many factors should be considered when choosing between the various methods. The choice depends largely on the
type of product and the form in which it was frozen. In some uses, it is difficult to provide sufficient shielding in the
dielectric thawer. The air-blast thawer might be at a disadvantage in the amount of cleaning required to keep bacterial
counts within the statutory limits. All the main methods described above can produce satisfactory results with blocks
of whole fish. Both heat conduction and electric methods fail with large fish. The electric methods give more uniform
thawing with compact block at small fish. 'Continuous thawers’ would appear to have advantage over batch thawers
for blocks at whole fish. For blocks of fillets, water thawing is unsuitable and block thickness probably should not
exceed 5 cm for thawing in humid air blast and between heated plates. The electric methods give good results with
blocks of fillets. Block upto 10cm thick have been thawed successfully dielectric thawer using only three generators.
Fillets, unprotected by the skin of the fish, are more susceptible than whole fish to deterioration on thawing,
particularly shrinkage, discolouration and dehydration. Pronounced thaw-rigor accompanied by shrinkage and loss of
fluid sometimes can be produced when thawing is rapid.

Unit 5: Quality changes during frozen storage

Chapter 1: Quality changes in fish during freezing & frozen storage-physical changes
5.1.1 Quality changes in fish during freezing

The physical changes which occur during freezing & storage of frozen products comprise crystallization of ice with
expansion of the volume, and desiccation starting from the surface of the frozen fish.

5.1.1.1 Ice formation

The crystallization of a ice is initiated when the temperature of the fish is lowered about –10C. At the same
time, concentration of various inorganic salts and organic components present in the fluid of the fish occurs and
consequently the freezing point falls. The temperature of the fish must be lowered further before additional water
freezes. There is also an increase in the volume of the fish when the water is converted to ice. At -30C about 70% of
the water is frozen. At -50C, about 85% is frozen; at-250C about 95% and at -500C to -600C, almost all the water in the
fish is frozen. The larger part of the water consequently freezes between -10C and -50C and it is the rate of cooling this
temperature interval determines the size of the ice crystals. The preserving effect of freezing however is due to the
extent of reduction in the rate of chemical and biological processes, when the temperature of the fish is lowered. The
43
crystallization or freezing of the water has certain un favorable effects. Rapid freezing, results in small ice crystals, and
the quality of quick frozen fish may be practically equivalent to that of fresh unfrozen fish.
Microscopic examination of frozen fish reveals that the changes occurring in freezing and thawing are caused
rather by irreversible changes in the colloidal state of the proteins induced by the increased concentration of
dehydrates in the unfrozen part of the fluid in the fish tissue. Small ice crystals are certainly a feature of quick freezing
whereas large ice crystals can be the result not only of slow freezing but also of considerable post mortem changes in
the tissue prior to freezing. Histological investigations on frozen fish revealed that when freezing fish with
considerable post mortem changes, the ice crystals destroy the sarcolemma more easily and grow large sizes causing
more histological changes to the tissue. The rate of freezing therefore has been considered of vital importance for
obtaining a high quality product. The DNA content in the drip also varied with rate of freezing. With more rapid
freezing, there is a drop in DNA content in the expressible fluid. In ultra rapid freezing, however, there is a rise in
expressible DNA, but probably due to different kind of cell damage.

5.1.1.2 Desiccation
Desiccation from the surface of the frozen fish takes place during freezing and storage because there is always
transport of water vapour from the fish to the evaporator or cooling coils. The greater the temperature difference
between fish and evaporator, and greater the velocity of the circulating air, the more moisture will be transported and
the more frost will be formed resulting in the decreased efficacy of the evaporator. The loss of water from the product
and the formation of snow on the evaporator coils are greater at high storage temperature, because the warmer air
will take up more moisture. Change in fluctuation in storage temperature influences the desiccation, loss of weight
and quality of the fish contributes to a poor appearance or results in “freezer burn”. The critical limit of dehydration
or rate of weight loss of frozen fish in cold storage is 50g/m2/ 24h.
Medium sized white fish stored in a rather dry room lost 7, 3.5 & 1.5% in weight per month at -9, -21 & -29oC ,
respectively. The air surrounding the frozen product is usually at a lower moisture vapour pressure than the product
and therefore acts as a sponge in removing moisture from the product. The loss of moisture results in the dehydration
of the flesh to a point where chalky and fibrous texture develops, discolouration takes place and off flavours develops
The phenomenon is also know as “Freeze burn”. When frozen fish is in contact with air, oxidation of fat or oil in the
fish takes place and these results in “rusting” or discolouration of the flesh and development of rancid odours and
flavours.
If the desiccation is pronounced, the fish surface may become dry and fibrous. In some cases the skin may change the
colour, several other factors influence the loss are weight, the kind of wrapping, it’s sealing and moisture transmission
characteristics. The rate of moisture loss or weight loss or water vapour transmission rate (WVTR) from fish in cold
storage is expressed in g/m2/day. The denaturation of the protein or reduced protein extractability is also due to the
electrostatic interaction between protein and fatty acid. Freezing increases toughness of fish muscle, which proceeds
progressively during subsequent storage. Toughness can not be measured in terms of percentage of soluble protein
in the cold stored fish. The proportion of fish muscle protein soluble in 5% Nacl solution was not correlated with the
development of toughness during cold storage at or below -200C. Assessment by a taste panel is time consuming and
inaccurate. Since fish muscle fibres are extremely short and interspaces. Instruments such as penetrometers have
limited usefulness. But by using a new method of treating the fish muscle with 1% formaldehyde, it was found that
the unfrozen fish muscle fibres were broken up & there by reducing the light transmission. Frozen fish showed a
higher proportion of intact fibres, letting through more light.

Toughness:

Toughness or changes in texture can be due to denaturation of proteins. The denaturation may be due to the
concentration of inorganic salts which results in damage to the flesh proteins or due to lipid hydrolysis and
myoglobin in the cells, are the main colouring pigments in fish flesh. The dark parts of the muscles contain much
more pigment than the light or white muscle.
5.1.1.3 Discolouration

In freshly caught fish the blood has a brick red color of oxy hemoglobin. This color gradually changes to the red
brown or brown color of met hemoglobin. Myoglobin suffers the same kind of changes. Frozen tuna and sword fish
may exhibit green and brown discolouration on looking. Uncooked fish meat contains three derivatives of
myoglobin. The pigment responsible for the pink colour in normal cooked meat of tuna is hemochrome, derived
44
from the reaction of myoglobin with non-heme constituents. Greening is due to pigments resulting from the
oxidation of hemochrome that occurs when the meat is unduly exposed to oxidative condition during and after
cooking. Greening of frozen sword fish may be related to take H2S produced by putrefactive bacteria. Proper
evisceration and removal of blood immediately after the catch reduce the risk of discolouration. The undesirable
discolouration in yellow fin tuna meat can be averted, if the fish is frozen at full rigor, stored at a temperature of -
230C to -270C, and defrosted by still air at 100C.

a. Carotenoid pigments:

Some fishes like salmon, and some crustaceans, contain carotenoid pigments in the flesh. The red color astaxanthin
tends to fade away during frozen storage. In crustacean, like lobster, pigment is limited to the surface of the meat,
the changes during frozen storage lead to yellow discolorations.

b. Fish oils:

The oils & fats naturally occurring in fish vary with species from colourless, as in herring through yellow to red, as in
salmon. Fat herring, with a layer of oil just under the skin, may develop a yellow color during poor storage condition
& which can be seen through the skin. Such fish is said to have “rusted”.

c. Black spots:

Shrimp, lobster etc, develop so called black spot when stored fresh. Bruises and rough handling increase the
occurrence of this discolouration, which is caused by an enzyme, tyrosinase acting on polyphenolic compounds,
present in free amino acids like tryptophan,tyrosine and form black 'melanin' pigments.

Flavor changes:

Systematic studies of flavor changes in frozen fish are lacking. Freshly processed fish, frozen and cold stored, is
generally considered first to lost it’s characteristic fresh fish flavour, becoming bland & insipid; there after it
develops flavors variably described as “cold stored” “Salt fishy’ etc.

Break down of connective tissue:

During some occasion, fish fillets are produced in which the muscle flakes will separate when the fish is filleted. This
defect, often called “gaping” appears to result from a breakdown in the connective tissue and associated
components which hold the muscle cells together. Although the main myofibrillar proteins, actin and myosin are
believed to remain largely intact, many of the cytoskeletal protein like connectin, nebulin and desmin are believed to
be degraded. The problem is, by and large, connected with freezing at sea and fillet production from frozen whole
fish.
5.1.1.4 Fat oxidation during freezing

The changes in lipids are directly and indirectly responsible for the quality deterioration in frozen seafoods. They
involve lipolysis, lipid oxidation, and interactions of the products of these processes with non-lipid components. The
endogenous fish lipases are relatively resistant to low temperatures and retain much of their activity in the frozen
tissues. Some of them may even be activated in the freezing process eg. by release from lysosomes. Fish lipids
undergo two main types of changes during storage of fish viz, hydrolytic changes and oxidative changes. These
changes result in rancidity.

Lipid hydrolysis

Lipases bring about hydrolysis of lipids producing free fatty acids and resulting in hydrolytic rancidity. Free fatty acids
trigger protein insolubilization and texture degradation in frozen stored fish.

45
Lipid oxidation

It is a very series problem in fish, in view of the highly unsaturated nature of fish lipids. The double bonds of
unsaturated fatty acids are highly susceptible to oxidation and this leads to the production of various carbonyls and
other secondary oxidation products, which impart the characteristic rancid off flavour to the product. These
products, besides producing off flavour reduce the shelf life and nutritional value of the product also. Some of them
are toxic in nature. These reactions are initiated by free radicals generated from unsaturated bonds, which start
chain reactions resulting in the production of various undesirable compounds like peroxides, hydroperoxides,
aldehydes, ketones etc. Finally, the free radicals form non-radical polymers, which terminate the chain reaction.

Oxidised lipids interact with proteins reducing the nutritive value of the proteins considerably. Melonaldehyde is one
of the major oxidation products and estimation of this compound by forming the thiobarbituric acid (TBA) complex is
the accepted method for monitoring the extent of lipid oxidation. In lipid oxidation, the first step leads to formation
of hydroperoxides, which are tasteless but can cause brown and yellow discolouration of the fish tissue. The
degradation of hydroperoxides gives rise to formation of aldehydes and ketones. These compounds have a strong
rancid flavour. Lipid oxidation primarily non enzymatic in nature, recently the involvement of microsomal enzymes
and lipoxygenase has been reported. This lipid oxidation takes place in fishes having more than 2% of the lipids eg.
fatty fishes.

Factors affecting the oxidation

1. Content and composition of unsaturated fatty acid

2. Oxygen availability
3. Light radiation
4. pH
5. Temperature
6. Moisture content

Content of pro and anti oxidant.

5.1.1.5 Factors limiting storage life

Protein changes: Fish proteins undergo permanent change during freezing and cold storage and the speed at which
this denaturation occurs depends upon temperature. At temperatures not very far below freezing point, -2oC for
example, serious changes occur rapidly; even at –10oC, the changes are so rapid than an initially good quality
product can be spoilt within a few weeks. The rate of deterioration due to protein denaturation, can be reduced by
storage at as low a temperature as possible.

5.1.1.6 Freeze Denaturation-I

Unfolding of molecules due to secondary reaction between the reactive groups of different proteins and other
components of fish muscle, leading to cross linking and formation of aggregates

 The proteins lose part of their solubility and reduced enzyme activity
 As a result of these changes significant deterioration of the functional properties of the fish meat may occur
 It is manifested by a decrease in water retention, gel forming ability and lipid emulsifying capacity worsening
of texture and the increased dryness of fish meat
 Fish that are stored for longer period becomes tough, chewy, rubber and fibrous
 Causative factors comprise the denaturing are

1. Catalytic effect of ice and inorganic salts

2. The binding of fatty acid

46
3. Lipid oxidation products

 Cross linking induced by formaldehyde generated in the muscle of some fish (Gadoid) results in the formation
of new covalent bonds in denatured products
 Ice crystals disturb the hydrophobic interaction in protein. Increase in concentration of salt in unfrozen food
and water decrease the hydration of protein as the concentrated inorganic ions compete for the water
molecules
 They participate in freeze denaturation of protein by catalyzing the hydrolysis and auto oxidation of lipids
 Free fatty acid decrease the solubility of myofibrillar protein and contribute to the formation of aggregates by
binding to proteins due to hydrophobic interaction
 Oxidised fatty acid and secondary products of autoxidation especially aldehydes are much more damaging to
protein solubility as they are able to form covalent bonds with reactive protein groups.

TMAO —>TMA + formaldehyde (HCHO)

So aldehydes is very reactive compound bind to some groups in protein side chain and form intra and intermolecular
methylene bridges. By removing water soluble substrates and enzyme system responsible for the degradation of
TMAO the undesirable protein changes can effectively be reduced.
5.1.1.7 Freeze Denaturation-II

Fat changes:

Fatty fish may become unpleasantly altered during cold storage but they can be protected to some extent either by
glazing or by packaging in plastic bags sealed under vacuum. These oxidation changes take place more rapidly at
higher temperatures and storage at a low temperature is an effective means of slowing the rate of spoilage by this
method.

Color changes:

The quality of fish is often judged by appearance, and color changes which are not otherwise significant can result in
fish being downgraded. The changes in the fish flesh which bring about these color changes are also retarded at
lower temperatures.

Dehydration changes:

Dehydration of the product is probably the major concern of the cold store operation and the rate of drying can be
linked with a number of factors in cold store design and operation.

When fish get badly dehydrated in cold storage, the surface becomes dry, opaque and spongy. As time progresses,
these conditions penetrate deeper into the fish until it becomes a fibrous, very light material. Visible effects of
severe dehydration on the surface of the fish are known by the term “freezer burn”. This is an unfortunate choice of
term since the effect is unlikely to result from freezing in a properly designed freezer, and appears only after periods
of storage in a cold store.

Frozen fish may dry slowly in cold storage even under good operating conditions. This is un desirable for reasons
other than the obvious one that the product will lose weight. Drying also accelerates denaturation of the protein and
oxidation of the fat in the fish. Even totally impervious wrappers used to protect the product do not give full
protection if the cold store operating conditions are favorable for desiccation within the pack. In-pack desiccation
prevails when there is some free space within the wrapper and temperature of the store fluctuates.

Weight loss during freezing will depend on: Type of freezer, freezing time, Type of product, Air velocity and Freezer
operating conditions. Cold Store Weight Loss Depends on Temperature, Temperature fluctuation, Humidity, Heat
transfer, Air flow over the product, Radiation effects of lighting, The product, Shape and size of the product and Type
of wrapper.
47
Chapter 2: Microbial growth at low temperature
5.2.1 Microbial growth at low temperature
Freezing of fish is done at – 400C and the frozen fish is further stored at – 180C. During freezing, 80 to 90%
of the Gram-negative bacteria die out and the residual bacteria cannot grow in the temperature of frozen
storage. So, during freezing preservation of fish, there is no bacterial spoilage. But, before cooking, the frozen fish
has to be thawed. During the thawing process the residual bacteria, which are predominantly Gram positive, can
cause spoilage of the thawed fish. Hence, frozen fish will have to be thawed within the shortest possible time. The
relation between microbial growth and temperature is still considered as complex phenomena governed by inter-
related factors such as substrate composition, freezing rate, microbial type.
As the temperature falls, bacterial growth rate is reduced and the lag period is extended until the minimum
temperature limit is reached when growth ceases. Some microbes will cease growing at 00C or even higher while
others continue to grow below the freezing point of food. Growth rate below 00C is very slow. The reported minimum
growth of microorganism varying from 100C to - 100C. But for practical cases, the lower limit of growth for bacteria,
yeast or moulds can take as -70C.

Most food stuffs contain low molecular weight compounds such as salts, sugars, etc. which decrease the
freezing point of water and lower the water activity. As more and more water is converted to ice with the fall in
freezing point, the water activity in the remaining unfrozen water is reduced. Hence, microorganisms as they
approache their minimum growth temperatures are subjected to two factors mainly, i.e. (i) the concentration of the
solutes in the unfrozen water which destroys the osmotic equilibrium of cell membrane leading to metabolic
diffusion and (ii) strong inhibitory action due to low water activity. Additional stress imposed on the cell prior to
freezing causes changes in the enzyme activities of microbial populations and reduces the efficiency of their
metabolic pathways and makes them more susceptible to low temperatures. This is of particular importance to food
such as fish, which is chilled prior to freezing. Hence, the ultimate effect of freezing is thought to be due to ice crystal
formation and solute concentration caused by the damage to the semi permeable properties of the cell membrane. At
temperature above -10°C, freezing occur only externally and a cell that could make osmotic adjustments and escape
death. On the other hand, below -10°C, cell membrane fails to act as an osmotic barrier to the proliferation of ice
already formed outside the cell resulting in intracellular ice formation and death. Damage to DNA is also involved.

When a food product having a mixed flora is exposed to low temperature, a small change in temperature
leads to substantial variation in the relative growth rates of the micro-organisms present. It has been conclusively
proved that Gram-positive bacteria particularly Gram-positive cocci are most resistant to deleterious action of
freezing. Yeasts and molds that are able to grow at relatively lower water activities are also a common component
in frozen foods. Cells those are not killed by freezing may be sub-lethally injured and such injury is reversible. The
repaired cells regain all the normal characteristics of the original cell within a few hours and this happens in the lag
period. Injured cells often exhibit increased nutritional needs, impaired membrane permeability, reduced resistance
to environmental stress (e.g. low pH) and increased sensitivity to inhibitory agents.

Qualitative changes in bacterial flora during freezing

Freezing imparts a selective action on the bacterial flora of fish and various bacterial species are affected at
different levels. But Gram-positive bacteria are more resistant to freezing and frozen storage than Gram
negatives. But among Gram positives, differing sensitivity to freezing is noticed. Bacillus, Lactobacillus and
Micrococcus are more susceptible to freezing than ‘Coryneform’ bacteria. Among the gram negatives, sensitivity to
freezing is almost comparable except the case of Vibrio species that are very much sensitive. Pseudomonas and
Aeromonas, Moraxella and Flavobacterium species are more resistant to the lethal action of freezing.

Effect of thawing
Generally frozen sea foods are considered highly stable and less prone to bacterial decomposition. But they
can never be considered as sterile and completely free from bacteria. If there is no contamination after thawing the
48
micro-flora on the thawed seafood will have only the bacterial population consisting of the freezing resistant species
only. This dormant population is inactive during the frozen storage period. But when the product is defrosted
(thawed) surviving bacteria are liberated and immediately begin multiplying resulting in the chemical breakdown of
the product.

During thawing process, bacteria start multiplying within the temperature limit characteristic for the different
species. The higher the external temperature the more favorable for outgrowth of most of the bacteria. Hence,
defrosting at a lowered temperature ensures a lower multiplication rate. Multiplication rate of bacteria will be
slower at low temperatures of thawing. As in the case of defrosting temperature, defrosting rate is also found to
have a decisive role in bacterial multiplication and the resulting spoilage. A slow rate of thawing allows the bacteria
to multiply considerably causing the spoilage of the product. Compared to other foodstuffs, there are greater
chances of spoilage of seafood during and after thawing because fish harbours a psychrophilic bacterial flora even
from the beginning. Though freezing and immediate storage kill as much as 50% of the psychrophilic organisms,
sufficient numbers remain to promote spoilage of thawed frozen product. When frozen fish is defrosted and kept in
the refrigerator, they undergo spoilage similar to that of unfrozen fish.

Unit 6: Cold stores and containers

Chapter 1: Construction of cold store
6.1 Construction of cold store

A cold store is any building or part of a building used for storage, at temperatures controlled by refrigeration at -
18ºC or lower.

Recommended Storage Temperature

Reducing the storage temperature can slow all the spoilage of fish due to protein denaturation, fat changes and
dehydration. The FAO Code of Practice for Frozen Fish recommends that frozen fish products should be stored at
temperatures appropriate for the species, type of product and intended time of storage. The International Institute
of Refrigeration recommends a storage temperature of -18ºC for lean fish such as cod and haddock and -24ºC for
fatty species such as herring and mackerel. The code also recommends that for lean fish intended to be kept in cold
storage for over a year, the storage temperature should be -30ºC.

Practical storage lives (PSL) of fish products

Storage life in months

Product
-18ºC -24ºC -30ºC

Fatty fish 5 9 >12

(glazed)

Lean fish (fillet) 9 12 24

Flatfish 10 18 > 24

Shrimp 5 9 12

(cooked/peeled)

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 6.1.1 Shape and size

Cold stores can be divided according to construction into single-storey and multi-storey buildings. They can be used
as production stores, bulk stores, distribution stores or retail stores. For a long time, the most appropriate shape was
a cube for which the ratio of surface to volume is a minimum. This resulted in multi-storey buildings, with a number
of disadvantages, e.g., costly foundations, heavy framework, congested handling areas.

The main considerations which have resulted in the appearance and success of single-storey buildings are cost
reduction together with mechanized handling techniques.

A single-storey cold store can be easily designed to meet the specific requirements of stacking and handling
equipment. Wall and roof constructions can be made lighter as they do not have to support the weight of the product
stored, as in a multi-storey building. The main disadvantages are the relatively large ground area covered and the high
ratio of surface to volume. The advantages however, normally override the disadvantages.
6.1.2 Insulation

The choice of insulation is very important as it accounts for a large proportion of the total construction cost. The
insulation material and thickness is also important from an energy point of view. Besides a satisfactory thermal
conductivity coefficient the insulation material should also be odour-free, anti-rot, vermin and fire-resistant and
impermeable to water vapour.

Typical insulation thickness for chill and cold stores utilizing different insulation materials

Calculated thermal conductivity Thickness

Insulation
(kcal/m hoC (mm)

Polystyrene 0.033 220

Styrofoam FR 0.030 200

Polyurethane 0.025 170

Vapour Barriers

Water vapour in the air gives rise to a pressure and together with the other gases present. The partial pressure
exerted by the water vapour is proportional to the quantity of vapour present and the vapour in the air will tend to
migrate from areas of high partial pressure to areas of low partial pressure. Hence, there is a tendency for moisture
in the ambient air to pass through the insulation of a cold store to the area of low partial pressure within. When this
vapour is cooled, it condenses and the build-up of ice will eventually affect the insulation properties of the cold store
wall and also weaken the structure of the wall or building.

Foundations and Frost Heave

Low temperature stores built directly on the ground may require special precautions to prevent the build-up of ice
below the cold store floor. The ice formation causes distortion known as “frost heave” (fig.) and in particularly bad
cases, it can lead to the complete destruction of the structure of the building. The conditions that give rise to frost
heave are rather complex, since they are related to the type and texture of the soil, the insulation properties, the
availability of moisture, the dimensions of the store, seasonal climatic variations and other factors. Two methods of
preventing frost heave are commonly used. The ground below the store can be heated either by a low voltage
electrical mat in the cold store foundation or by circulating a heated liquid such as glycol through a pipe grid built

50
into the foundation. The heat for the glycol is usually obtained from the compressor hot gas through a heat
exchanger.
6.1.3 Air ingress

Outside air entering the store adds heat and moisture. This moisture will be deposited as frost on any cold surface
and will eventually cover the surface of the cooler. Excessive air exchange should be prevented to keep the cold
store temperature steady and reduce the frequency of defrosting. Small air locks have been used to prevent the free
flow of air in and out of the store.

A curtain of air blown downward or from the side of the doorway can reduce the exchange of air when the door is
open. These air curtains, as they are called, can be a useful aid when the door is opened for short intervals. However,
they are often abused and doors are often left open for long periods.

Floors

The ground loads from a cold store are in the order of 5500-8000 kg/m2. This consists of static loads due to
merchandise, structure and concentrated rolling loads transmitted by e.g., forklift trucks and other handling
equipment. It is of importance that those loads are investigated in detail for each special project. Most commonly
the floor wearing surface is a concrete slab cast on the floor insulation with a thickness of 100-150mm. In cases
where intensive traffic is foreseen a special hardwearing top-finish is recommended. Before casting the wearing
surface, the floor insulation should be protected by bituminous paper or plastic sheeting, the function of which is
twofold.
6.1.4 General layout

A single-storey building can either have one single room or it can be divided into a number of rooms. Normally all
the rooms are operated at the same temperature, for fish preferably in the range of –24o to -30oC. Most stores are
built at a higher level than the surrounding yard with a special loading ramp at one or more sides. The loading ramp
level corresponds to the height of the most commonly used vehicles.

The engine room should be as close as possible to the position of the air cooling equipment within the store.

Modern large or medium cold stores are built as one-storey buildings designed for mechanical handling, e.g., forklift
trucks and automatic stacker cranes. Manual handling is, however, still used for most small-sized stores.

A cold store can be built as an ordinary building using conventional building material, such as bricks, concrete or
concrete precast sections to which a vapour barrier and insulation is fitted internally. Modern insulation material, in
particular polyurethane, has a strength that can be utilized structurally. Today, this is used for panel designs suitable
for all sizes of cold rooms from 20m3 to 250 000m3. Factory made insulation panels are delivered to the site
complete with a vapour barrier and internal cladding, thus reducing the site work to a minimum. There are two basic
principles for panel-built cold stores. A common system has an external structure and cladding with wall insulation
inside of the columns and the insulated ceiling hanging from the outer roof structure.

The panels normally used in these systems are either polyurethane or polystyrene insulated panels with or without
frames. They are manufactured as sandwich panels, one face being the vapour barrier of light-gauge galvanized steel
sheet and the other face being the internal finish of plastic-coated galvanized sheet or aluminum sheet. A decorative
external cladding is erected on the outside of the columns. The roof insulation is constructed as a suspended ceiling.
The roof panels are, in principle, the same as the wall panels, but are sometimes equipped with wooden frames.

The latest development are panels with a rib profile on the external face of aluminum – with polyurethane foam
insulation and an internal face of low profile corrugated aluminum. The space between the panels is filled with one
component polyurethane foam. The internal cladding is sealed with a PVC strip. One component foam is also used to
join wall panels to roof panels and seal around doors, etc, maintaining good insulation properties throughout the
building. With this design both the insulation and external vapour barrier are entirely sealed units enveloping the

51
whole building. This means that losses via heat bridges or air leakage are completely eliminated, which gives
practical insulation properties closer to theoretical values than normally expected.

Chapter 2: Refrigerated containers

6.2.1 Refrigerated Containers

In a refrigerated transport container or insulated vehicle it is difficult to maintain a low and stable produce
temperature. Added to this, the effect of fans, the opening of the door and the presence of workmen is
proportionately greater in refrigerated transport than in a large, well-designed storage chamber. Under operating
conditions the best temperature obtainable is approximately -180C to -150C.

It follows that produce should be kept in transport vehicles and containers for as short a time as possible. Very often
parked containers are used for cold storage to avoid the expense of constructing a cold store. However one has to
keep in mind that parked transport containers and vehicles are not suitable for use as stores for frozen produce
except for very short periods and in emergencies. There is such greater risk of loss of quality through temperature
fluctuation and desiccation in refrigerated containers than in large, well-designed and well-operated cold store.
Storage in transport containers is also very expensive when compared with properly designed cold stores.

The Cold Chain

Temperature is the single most important factor influencing fish spoilage. The cold chain is a temperature chain. It
means 00C for fresh fish and -200C for frozen fish from the producer to the consumer. Temperature rise and
temperature fluctuation will accelerate quality deterioration and it should to be avoided. Especially sensitive to
temperature abuse are fresh, live and lightly cured (e.g. smoked fish) products. These should be kept at 0-10C, and
their temperature should never rise above 30C.

Temperature fluctuations accelerate the reformation of small ice crystals into larger crystals damaging the cells.
Also, desiccation will increase because of temperature fluctuations.

The insulated transport vehicles and containers shall be provided with adequate quantities of coolant or good
mechanical refrigeration. Refrigeration requirements vary considerably depending on the duration of transportation,
the temperature at which a product has to be maintained, the quality of insulation material, the dimensions of a
container and the temperature of the environment.
6.2.2 Loading

The transport vehicle has to be pre-cooled for several hours prior to loading. With pre-cooling a large rise of product
temperature will be avoided. Furthermore frozen and chilled fish should not be left outside for a considerable length
of time and exposed to high temperatures. To avoid such exposure fast loading and unloading is required.

Transit Points

This shipper should take into consideration that transit points like airports often do not have proper refrigeration
facilities, or because of mismanagement, chilled consignments are not stored in refrigerated rooms while in transit.
When no refrigeration facilities are available, more coolant should be used. Alternatively the consignment might be
re-iced during transit. The best way to avoid temperature abuse at transit points is to have good organization and
planning.

Temperature Monitor

To check on temperature abuse, continuously working monitors have been developed. These give proof of proper
temperature control or abuse during road transport, at airport terminals and so on. Because of the financial risks
involved it is necessary to use such monitors especially with large or high value consignments.

52
Chapter 3: Good handling and shipping practices
6.3.1 Fresh fish

The important factors in the distribution of fresh fish are speed, cleanliness and the need to keep the fish well
chilled. The fish must not be handled repeatedly.

Everything likely to come into contact with the fish has to be clean. Bacteria may be picked up from dirty surfaces,
tools, uniforms and hands. This will accelerate, fish spoilage and may endanger public health. Fish boxes, trucks,
loading platforms have to be cleaned and disinfected before use.

For shipping fresh fish, refrigerated transport is required. Temperature is the single most important factor influencing
fresh fish spoilage. Chill fresh fish with ice immediately and keep it cool. Mix the fish and ice well. Ice must always
remain in sufficient quantities for effective chilling and to separate the fish from each other and from the supporting

surfaces. The air temperature in the container should be 2-30C to allow the ice to melt slowly.

Fresh Fish Spoilage

 Coldwater fish spoils faster than warm water fish.

 Fatty fish spoils faster than lean fish.
 Smaller sized fish spoil faster than larger fish of the same species.
 Round fish spoils faster than flat fish.
 Shark spoils faster than lean fish.

Fresh Fish Fillets

Direct contact between fillets and ice is not recommended. Water will soften the texture and affect appearance.
Separate the ice from the fillets e.g. by using plastic sheeting. Be sure to chill the fillets properly before packing.
Chilling after packing is difficult because of the tightly packed mass of fish flesh.

Frozen Fish

For shipping frozen fishery products refrigerated transport is required. The cold chain has to be maintained at
all times. The normal working temperatures in modern cold stores for quick frozen foodstuffs are -30 to -250C. Retail
cabinets, in which the product should be stored for no more than a few weeks, usually operate at about -180C or a
little lower.

Do not try to freeze fresh produce or try to lower the temperature of frozen fish when shipping. Prior to
shipment always check the product temperature with a thermometer. Frozen fish should be handled with care.
Rough handling, such as throwing, will cause the frozen flesh to crack, damage the skin and fins and these defects
will inevitably show up when thawed.

Unit 7: Packaging methods

Chapter 1: Packaging for fresh fish handling
7.1.1 Packaging for Fresh Fish Handling

To maintain the quality of fresh fish, use of good fish boxes and packaging materials is required. Specially designed
and constructed fish boxes are now widely used, such as insulated containers for air shipment. Appropriate
packaging also makes handling easier, which reduces labour costs.

Important Requirements for Fish Boxes and Containers

 They must be of a suitable size for the range of fish to be handled or the products to be put into them.

53
  They should be of a convenient size for manual handling or lifting by mechanical equipment.
 They should be stackable such that the weight of the containers on top rests on the containers underneath
and not on the fish.
 They should be constructed of impervious non-tainting materials.
 They should be easy to clean.
 They should provide drainage for the melt –water.

Polyethylene Fish Boxes

High density polyethylene is most commonly used for fish boxes as it is strong and light in weight and easy to clean.
Polyethylene fish boxes are very popular onboard fishing vessels.

Disposable Fish Boxes

The disposable boxes are commonly used. These hold up to approximately 25 kg of fish or fish plus ice. They may or
may not be insulated. Insulated boxes include fiberboard boxes with polystyrene liners.Non-insulated fish boxes are
normally fiber board cartons, waxed or otherwise water-proofed. A road transport, disposable packaging can be a
water-proofed carton with or without insulation depending on whether the transport vehicle is refrigerated or
not. Packaging for air transport, has to be light, strong and leak proof, may consist of a heavy duty water-proofed
corrugated carton with two polyethylene liners and insulating material between the liners.
7.1.1.1 Moulded polystyrene Fish Box

Moulded polystyrene fish boxes are commonly used for delivery of chilled fish and frozen fish. In a typical range the
wall thickness varies with the box size, for example a 6 kg capacity box has a 15 mm thick wall, a 10 kg box a 19 m
wall, a 25 kg box a 25 mm wall. The major disadvantages of polystyrene boxes are their lack of strength. This limits
their size and use. They are easily damaged or broken by rough handling. Polystyrene is difficult to clean. Polystyrene
boxes are difficult to re-use, and are normally non-returnable .

Use of Insulating Linings

Wooden or fiber board boxes can be insulated with polystyrene sheeting cut to the required dimensions.
Alternatively waxed fiber board which has good water resistance can be used. The empty box can be shipped in a
knocked down form. This is cheaper than using moulded polystyrene boxes.

Light-Weight Insulated Packaging

A recent development in light-weight insulated packaging makes use of the ability of metallised surfaces to reflect
practically all (97%) radiant heat and so to reduce the thermal insulation thickness. The thermal guard packaging
system consists of a metallised plastic bag in which the produce is packed, a bubble-pack insulating wrapper,
provides the insulation. The thermal guard bag can be sealed airtight and can thus be used for frozen fish.

Returnable Versus Disposable Containers

The environmental pollution, authorities in many countries try to reduce the use of disposable containers. The
returnable containers are the freight costs for returning empty containers, the logistics involved and the high
investment in good quality containers. Using a ‘knock –down’ returnable container will reduce return freight costs.
The returnable containers are to be preferred because in the long run these will then be cheaper and generate less
trash.

7.1.1.2 Refrigerants and Coolants

Ice should be properly mixed with the product and applied in sufficient quantities to last for the duration of the
shipment. When fish are in close contact with each other or with a smooth impervious surface such as the inside of a
box, anaerobic bacteria producing offensive odors may develop. The air temperature in the container should be 2-3ºC

54
to allow the ice to melt slowly. Melt-water keeps the fish well chilled and moist. The melt-water also helps to wash
away the products of spoilage. The melt-water should be drained away from the produce.

Alternative refrigerants to wet ice are dry ice and gel ice. Dry ice is solid carbon dioxide. Evaporation of the dry ice
gives the cooling effect. Because of its low temperature (-78.9ºC), dry ice should not be in direct contact with chilled
fish, to avoid cold burns.

Gel ice is made by freezing a water based gel. The advantage of gel ice is that all the water is bound with no danger
of water leakage during thawing. It is often claimed that gel packs release the ‘cold’ more slowly and equally than
other coolants so that they are superior in maintaining chilled temperatures.

Chilling air cargo

With air cargo often preference is given to dry ice or gel packs, as these do not cause leakage. However the
use of dry ice is subject to restrictions because it expands from a solid form into a gas and it may ‘drive-out’ oxygen.
This obviously poses a hazard to the safety of airplanes. The international identification number for dry ice is UN
1845.

7.1.1.3 Guidelines for air shipment of sea food

 Boxes, cartons or other outer packaging materials must be multiwall, was impregnated or waterproof coated
on both inside and outside. Gusseted corners must be on both top and bottom with a top that extends fully
over to the bottom.
 Outer packaging must be banded in two directions.
 Use dry-ice or gel ice preferably.
 Product must be sealed in a polyethylene bag with wet ice a minimum 4 mm gauge polyethylene bag or liner
is required.
 Use an absorbant pad.
 Styrofoam boxes are to be protected by wet lock cartons.
 The cartons must clearly indicate the airway bill number, name of the shipper, consignee’s name and
address, plus marking like “This Side Up” and “Perishable Fresh Sea foods”.
 With dry ice follow the IATA Dangerous Goods Regulations to declare use and amount of dry ice.

Chapter 2: Packaging of frozen fishery products

7.2.1 Introduction
The factors that must be taken into account while selecting a package for frozen foods are, protection of the
contents from

 atmosphere oxygen
 loss of moisture
 flavor contamination
 entry of microorganisms
 mechanical damage
 exposure to light

In addition to the above, the materials used must have a high heat transfer rate to facilitate rapid freezing. The
package shall have an aesthetic appeal and promotes sales of product and hence a well designed package is sometimes
called the ‘silent salesman’. Quick freezing food require transparent, moisture vapour proof, wet strength, tasteless
and odourless wrapping materials.
Modern frozen food processors can select from a wide range of protective materials such as variety of papers (Kraft
papers coated with wax or plastic and laminated papers), films (moisture proof, heat sealable cellophane and various
thermoplastic films such as saran, mylar, pliofilm and plastic wrap) and foils (heavy aluminum foil) in designing the
package.

55
The films are transparent, lightweight, resistant to puncture and flexible at low temperatures, free from off odours
and off flavours, non-toxic, heat sealable and have a low moisture vapour transmission rate.
7.2.1.1 Primary packages
They are three forms a plastic packaging:

1. Flexible films,
2. Semi rigid packages,
3. Rigid packages,

It is estimated that more than 600 flexible film combinations are commercially available today. Terms frequently
used in describing flexible film packages are coatings and laminations.
7.2.1.2 Flexible films
1. Flexible films

A variety of flexible films is used in frozen food packaging as bags, pouches and over wraps. Almost all of these
films are made from organic polymers. Some of the natural polymers such as starch, cellulose and rubbers are also
useful in food packaging.

Polyethylene (PE): Polyethylene is a long chain polymer of ethylene, a gas derived from natural gas or petroleum. It
is an useful film in frozen food packaging. It is low in cost, has excellent clarity and gloss, is resistant to tearing, has
relatively low WVTR, is flexible at low temperatures and is heat sealable. Because of the last property, it is frequently
used as coating or as layer in a laminate with heat sealable films. Important properties such as heat seal ability,
moisture resistance and flexibility depend on the density of the polyethylene resin from which it is made. Polyethylene
is classified into three density ranges and expressed as g/cc as follows.
(1) Low – 0.910 to 0.925
(2) Medium – 0.926 to 0.940
(3) High – 0.941 to 0.965
Most polyethylene films used in frozen food packaging are in the low and medium density ranges.
Nylon (Polyamide PA): It is a group of long chain polymers with re occurring amide groups as a part of the chain. Nylon
films have excellent properties of toughness, tear and breaking strength. They are useful both as single films and in
many laminates.

Plio films: It is the trade name for rubber hydrochloride film. It was the first transparent film which could be heat
sealed.

Polypropylene (PP): It is relatively new film in frozen food packaging, but its clarity, strength at low temperatures and
low WVTR make it useful in laminates, and in coating paper or aluminum foil. It is heat sealable, when it is heated with
a variety of coatings.

Cellophanes: Cellophane is made from regenerated cellulose. When it is coated with other plastic resins such as
polyethylene or saran, a wide variety of cellophanes result, each designed for a specific use. These make excellent
packaging films. They are strong, moisture proof, heat sealable and flexible at low temperatures.

Poly vinylidene chloride (PVDC): Polymers of vinylidene chloride are better known as saran and are available not only
as films, but as water based emulsions in which form they are frequently used as coating resins for paper substrates.
Saran is superior to polyethylene as a water vapour barrier material and has low oxygen permeability. The film is not
heat sealable. The film is excellent for use as a wrap around odd shaped meat (shrink wrapping).

Polyesters (PES): This strong film is well known under the trade name of Mylar. It is frequently used in lamination with
other films, especially with polyethylene to make pouches which can be filled and heat sealed. These laminates are
now used for boil in the bag pouches. Polyester films are strong, durable, transparent, non-toxic, inert and easily
stretched. They also have the advantage of being flexible at very low temperatures (-710C). They are sometimes known
as nylon type films and because of their roughness,they are useful in packaging sharp pointed, irregular foods such as
lobsters.
56
2. Plastic Pouches
The pouch is usually made from a laminate of polyethylene, nylon and Mylar films. One of the weakest links in
plastic bags is the seal, both from the stand point of vapor leakage and that of bag breakage at the seal.
7.2.1.3 Shipping container

Individual packages must be further packed in a properly engineered container for shipping and storage. Corrugated
containers are the most widely used form of containers for shipping frozen foods. Wooden crates and foam
polystyrene shipping containers are also used. The use of a corrugated sheet provides exceptional crush resistance.
It also has a high capacity for absorbing shock.

Corrugated fiber board is made of fluted (grooved) sheet glued between two liners. Four types of corrugated
board are commercially used and they are A,B,C and E flute. The difference lies in the number of flutes and board
thickness. ‘A’ flute -35 flutes per inch and 3/16 inch thickness; ‘B’ flute -50 flutes per inch and 1/8 inch thickness; ‘C’
flute -41 flutes per inch and 5/32 thickness; ‘E’ flute -90 flutes per inch and 5/64 inch thickness. Each type has some
specifications and advantage over the others.

Though various coating methods have been used, the most recent technique is called ‘curtain coating’, wherein
the coating formulation is applied by a machine designed for that purpose. The continuous film coating acts as an
effective barrier, not only against free water, but also against water vapor, gas, grease, oil, and other foreign
substances. It is necessary to use a coated shipping container, if there is a possibility of the container being exposed
to high humidity or excessive moisture conditions during handling.

The foam polystyrene container for shipping fresh frozen foods is relatively new. It has the advantage of being
water resistant and having excellent insulating properties. After the frozen product is placed inside, the foam lid is
applied, and the container sealed off. It has the disadvantage of being somewhat more costly and hardens to handle.
The empty containers are bulky to handle and store. However, because of its light weight and insulating properties,
the polystyrene container is particularly adaptable to the air transit of frozen foods.
7.2.1.4 Shrink packaging
This type of packaging was introduced by the Cryovac company in 1948 as a protective package for frozen
poultry. The saran-type film which was first used for this kind of packaging offered excellent protection against
freezer burn, and loss of weight by desiccation. In the cryovac process, the whole bird, or a large piece of red meat is
inserted into a plastic bag, air is removed by vacuum so that the bag is drawn in tightly against the bird, the bag is
sealed by twisting and clamping and film is shrunk into a skin-tight wrap by dipping the package into hot water
(about 900C)
7.2.1.5 Requirements for packaging materials

The requirements for packaging materials for frozen foods to be measured include

1. tensile strength and elongation

2. tear strength
3. impact strength
4. stiffness
5. bursting strength
6. fold endurance
7. compression
8. grease resistance
9. water vapor transmission.

A packaging material may be water proof without being water vapor proof. A water proof material will hold water
and will not go to pieces in water. A water-vapor proof material prevents vapor from passing through it.
Packaging Machinery
The packaging operation can be divided into four major steps

57
 1. Forming the package
2. Filling the package
3. Closing and sealing the package
4. Placement in the shipping container

Chapter 3: Vaccum packaging and modified atmosphere packaging and storage

7.3.1 Vaccum packaging

 It is a viable technology in seafood processing.

 It is wide spread in the meat industry for fresh, processed and cured products but has faced difficulties in its
application to seafood.

Condition in the vaccum package

It is brings several changes

1. A drastic reduction in O2 content

2. An increase in CO2
3. Increase in acidity

Advantages:

 It increase the shelf life of fish (ie., It is very good barrier to O2)
 Variety of spoilage organisms is substantially decreased
 Microbial growth is prevented entirely (Partially)

Packaging Materials

Light, water, heat, O2, bacteria and are consider to be an affecting factors in this packaging.

Light

The length of the exposure, the wave length and intensity of light and permeability of the package to the light affect
nutrients.

Temperature

This type of packaging material greatly affect the rate at which heat i.e. transferred to and from the food.

This is important for both refrigerated and frozen seafood.

Insulating containers like Styrofoam food trays greatly reduce the transfer of heat of the fish.

Oxygen

Excessive amount of O2 promote seafood deterioration through the oxidation of fats, the degradation of some
amino acids and the destruction of certain vitamins.

The effects of O2 are related to the atmosphere pressure,humidity and temperature.

Exposure of the package contents to O2 may occur through loose closure, puncture and or diffusion through the
packaging materials itself. Permeability of O2 also affected by the moisture and temperature.

Disadvantages of vaccum packing

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  Doesn’t ensure free from microbial activity.
 The O2 impermeable materials required are relatively expensive and appropriate equipment is needed.
7.3.2 Modified atmospheric packaging and storage

 In 1932, conye reported the beneficial effects of CO2 in inhibiting the growth of bacteria from fish.
 They have also been applied to other storage systems, namely CO2 saturated sea water and CO2 enriched
atmospheres.
 Increased storage life of fresh fish and shell fish using refrigerated sea water saturated with CO2 become
evident.
 The results appear to apply to a wide variety of species including fatty species of salmon, low fat rock fish and
shrimp.
 When the air surrounding the fish is changed in composition usually by enriching it with CO2 and the container
then sealed the atmosphere is called “modified”.
 Fish and shell fish from modified atmosphere storage have lower bacterial counts, greater acidity level,
reduced trimethylamine formation and lower NH3 production.
 There is also residual and positive effect on the keeping quality after removal from the CO2 enriched
environment.
 Temperature abuse results in bacterial spoilage and possible development of toxins. The accumulation of
unpleascent odour in the package owing to the activity of anaerobic organisms.
 The more serious problem is the threat of C. botulinum activity

The odour are not the usual type of “fishy” odours produced by sea food because of the circulation of fresh gas
removes them.

Unit 8: HACCP
Chapter 1: Hazard analysis and critical control point- A detailed approach
8.1.1 Introduction to the codex Alimentarius commission

The Codex Alimentarius Commission (CAC) was established by FAO in 1961. CAC is an intergovernmental body with
158 member Governments as on 31st August, 1997. The Codex Alimentarius (‘Food Code’ or ‘Food Law’ in Latin) is a
collection of food standards, codes of practice and other recommendations presented in a uniform way. Codex
standards, guidelines and other recommendations ensure that food products are not harmful to the consumer and
can be traded safely between countries. The Codex Alimentarius has resulted in evaluation of the safety of over 760
food additives and contaminants and setting of more than 2500 maximum limits for pesticide residues and more
than 150 for veterinary drug residues.

CAC has adopted the guidelines for the application of the Hazard Analysis Critical Control Point (HACCP) system
through its committee on Food Hygiene. It has recognized HACCP as a tool to assess hazards and establish control
systems that focus on preventive measures instead of relying primarily on end product testing.

The Uruguay Round of Multi lateral Trade Negotiations which concluded in 1994 established the World Trade
Organization (WTO) to replace the General Agreement on Tariffs and Trade (GATT).

1. The Agreement on the Application of Sanitary and Phyto Sanitary Measures (SPS Agreement) The SPS
Agreement confirm the right of WTO member countries to apply measures necessary to protect human,
animal and plant life and health.
2. The Agreement on Technical Barriers to Trade (TBT Agreement). The TBT Agreement basically provides that
all technical standards and regulations.

8.1.2 Work of the codex alimentarius commission

The work of the codex Alimentarius Commission – including the guidelines for the application of HACCP system – has
become reference for international food safety requirements. The application of the General Principles of Food

59
Hygiene and Good Manufacturing Practices (GMPs) allows the producer to operate with in environmental conditions
favourable to the products.

The main objectives of the Codex General Principles of Food Hygiene are

1. Identify the essential principles of food hygiene, applicable throughout the food chain to achieve the goal of
ensuring that food is safe and suitable for human consumption
2. Recommended an HACCP based approach as a means to enhance food safety
3. Indicate how to implement those principles and
4. Provide guidance for specific codes which may be needed for sectors of the food chain, processor, or
commodities and to amplify the hygiene requirements specific to those areas.

8.1.3 History of HACCP

Dr.Deming and others developed total quality management (TQM) systems for Japanese products in the
1950’s.NASA wanted a “Zero defects” programme to guarantee the safety of the foods that astronauts would
consume in space. Pillsbury presented the HACCP concept publicly at a conference for food protection in 1971. The
use of HACCP principles in the promulgation of regulations for low-acid canned food was completed in 1974 by the
United States Food and Drug Administration (FDA). In the early 1980’s the HACCP approach was adopted by other
major food companies.
The International Commission on Microbiological Specifications for Foods (ICMSF) and the International Association
of Milk, Food and Environmental Sanitaries (IAMFES), have recommended the broad application of HACCP to food
safety.

ADVANTAGES OF HACCP

The HACCP system, as it applies to food safety management, uses the approach of controlling critical points in food
handling to prevent food safety problems. The system, which is science based and systematic, identifies specific
hazards and measures for their control to ensure the safety of food. The application of the HACCP system can aid
inspection by food control regulatory authorities and promote international trade by increasing buyers’ confidence.

Guidelines for the application of the HACCP system

Management commitment is necessary for implementation of an effective HACCP system. During hazard
identification, evaluation and subsequent operations in designing and applying HACCP systems, consideration must
be given to the impact of raw materials, ingredients, food manufacturing practices, role of manufacturing process to
control hazards, likely end use of product, categories of consumers of concern, and epidemiological evidence relative
to food safety.

The intent of the HACCP system is to focus control at CCPs by all means.

8.1.4 Principles of the HACCP system

1. Conduct a hazard analysis

2. Determine the critical control points (CCPs)

3. Establish critical limit(s)

4. Establish a system to monitor control of the CCP

5. Establish the corrective action to be taken when monitoring indicates that a particular CCP is not under
control

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 6. Establish procedure for verification to confirm that the HACCP system is working effectively

7. Establish documentation concerning all procedures and records appropriate to these principles and their
applications

Logic sequence of application of HACCP

Assemble HACCP

Describe product

Identify intended use

Construct flow diagram

On-site confirmation of flow diagram

List all potential hazards conduct a hazard analysis, consider control measure

Determine CCPs

Establish critical limits for each CCP

Establish a monitoring system for each CCP

Establish corrective actions

Establish verification procedures

Establish documentation and record keeping

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 8.1.5 The HACCP team

Team composition:

When selecting the team, the coordinator should focus on:

o Person who will be involved in hazard identification

o Person who will be involved in determination of critical control points
o Person who will monitor CCPs
o Person who will verify operations at critical control points
o Person who will examine samples and perform verifications procedures

Knowledge required:

Selected personnel should have a basic understanding of:

o Technology and equipment used on the processing lines

o Practical aspects of the food operations,the flow and technology of the process
o Applied aspects of food microbiology and HACCP principles and techniques

Product description:

1. Product name (common name) or group of product names (the grouping of like products is acceptable as
long as all hazards are addressed)
2. Important end-product characteristics: properties or characteristics of food under review those are required
to ensure its safety (e.g. aw, pH, preservatives)
3. How the product is to be used (i.e. ready to-eat, further processing required, heated prior to consumption)
4. Type of package, including packaging material and packaging conditions (e.g. Modified atmosphere)
5. Shelf-life, including storage temperature and humidity if applicable
6. Where the product will be sold ( e.g. Retail, institutions, further processing)
7. Labelling instructions (e.g. Handling & usage instructions)
8. Special distribution control ( e.g. Shipping conditions)

8.1.6 Form. 2 Product ingredients and incoming material

List the product ingredients and incoming materials (including raw material, product ingredients, processing aids,
and packaging materials) that are used during the manufacturing process. This exhaustive listing is required for
proper identification of all potential hazards that could apply.

Flow diagram: - (will identify the important process steps)

o All ingredients and packaging used (biological, chemical, physical data)

o Sequence of all process operations (including raw material addition)
o Time/ temperature history of all raw materials and intermediate and final products, including the
potential for delay.
o Flow conditions for liquids and solids
o Product cycle/rework loops
o Equipment design features

Plant schematic:

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This plan should aid in the identification of any areas of potential cross-contamination with in the establishment. The
plant schematic/floor and equipment layout should be considered in detail and assessed. Data may include but is not
restricted to:

o Personnel routes
o Routes of potential cross – contamination
o Area segregation
o Flow of ingredients and packaging materials
o Location of changing rooms, wash rooms, lunch rooms and hand-washing stations

8.1.7 Hazard analysis: (Principle 1)

Hazard will vary among firms making the same products because of differences in:

 Source of ingredients
 Formulations
 Processing equipments
 Processing and preparation methods
 Duration of process
 Storage conditions
 The experience, knowledge and attitude of personnel

Therefore hazard analysis must be done on all existing and new products. Changes in raw materials, product
formulations, processing or preparation procedures, packaging, distribution and/or use of the product will require
review of the original hazard analysis.

Examples of biological hazards: Viruses

Bacteria (Spore forming): Hepatitis A & E

Clostridium botulinum Norwalk virus group

C.perfringens Rotavirus

Bacillus cereus Protozoa & parasites:

Bacteria (Non-spore forming): Diphyllobolthriumlatum

Brucella abortis Entamoeba histolytica

B.suis Giardia lamblia

Campylobacter spp Ascaris lumbricoides

Cryptosporidium parvum Taenia solium

(E.coli 0157; H7, EHEC,EIEC, Taenia saginata

TEC, EPEC) Trichinella spiralis

Listeria monocytogenes

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 Salmonella spp.

S.typhimurium S. enteriditis

Shigella spp .

Staphylococcus aureus

S. pyogenes

Vibrio cholerae

V.Parahaemolyticus

V.vulnificus

Yersinia enterocolitica

Examples of chemical hazards:

Hazards Polychlorinated biphenyls (PCBS)

Naturally occurring chemicals Agricultural chemicals:

Allergens  Pesticides
 Fertilizers
Mycotoxins (eg.flatoxin)  Antibiotics
 Growth hormones prohibited substances
Scombrotoxin (histamine)  Direct
 Indirect
Ciguatoxin

Biogenic toxins

 Paralytic shellfish
poisoning (PSP)
 Diarrhoetic shellfish
poisoning (DSP)
 Neurotoxic shellfish
poisoning (NSP)
 Amnesic shellfish
poisoning (ASP)
 Phyrolizidine alkaloids
 Phyrohaemagglutinin

Toxic elements and compounds from packaging materials

- Lead - Plasticizers

- Zinc - Vinyl chloride

- Cadmium - Mercury - Printing/coding inks

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 - Adhesives
- Arsenic
- Lead
- Cyanide
- Tin
F ood additives vitamins &
Minerals contaminants

- Lubricants

- Cleaners

- Sanitizers

- Coatings

- Paints

- Refrigerants

- Water or steam treatment

chemicals

- Pest control chemicals

8.1.8 How to conduct a hazard analysis

After listing all the hazards (biological, chemical or physical) that may be reasonably expected at each step from
primary production, processing, manufacturing and distribution until the point of consumption, the HACCP team
should assess the potential significance or risk of each hazard by considering its livelihood occurrence and severity.
The estimate of the risk of a hazard occurring is based upon a combination of experience, epidemiological data and
information in the technical literature. Severity is the degree of seriousness of the consequences of a hazard if the
hazard is not controlled.

Hazard addressed under the HACCP system must be of such a nature that their prevention, elimination or reduction
to acceptable levels is essential to the production of safe foods. Hazards of a low probability of occurrence and a low
severity should be addressed under the HACCP system but may be addressed through the good manufacturing
practices (GMPs) contained in the codex general principles of Food Hygiene.

8.1.9 Hazard assessment

Severity:
Severity is the magnitude of a hazard or the degree of consequences that can result when a hazard exists. Disease
causing hazards can be categorized according to their severity. One system uses the categories of:

 High (life-threatening ) ex. include illness caused by C.botulinum, S.typhi, Listeria monocytogenes, E. coli 0157
H7, V.cholerae, V. vulnificus, paralytic shell fish poisoning and amnesic shell fish poisoning.
 Moderate (severe or chronic) eg. include illness caused by Brucella sp. Campylobacter sp. Salmonella sp,
Shigella sp, Streptococcus sp, Yersinia entrocolitica, hepatitis A virus, mycotoxins, Ciguatera toxin
 Low (moderate or mild) – ex. include illness caused by Bacillus sp. C.perfringens, Staphylococcus aureus.
Norwalk virus, most parasites, histamine – like substances and most heavy metals that cause mild acute
illnesses.

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 Risk is a function of the probability of an adverse effect and the magnitude of that effect, consequential to a hazard(s)
in food. Degrees of risk can be categorized as high (H), Moderate (M) low (L) & negligible (N).

8.1.10 Critical control points: Determination (Principle 2)

The determination of CCPs is the 2nd principle of HACCP. The codex guidelines define a CCP as a step at which
control can be applied and is essential to prevent or eliminate a food safety hazard or reduce it to an acceptable level.
The determination of a CCP in the HACCP system can be facilitated by the application of a decision tree such as that
included in the codex HACCP system and guidelines for its application which indicates a logical reasoning approach.

Proceed to the next identified hazard in the described process

** Acceptable and unacceptable levels need to be defined within the overall objectives in identifying the CCPs of
the HACCP plan.

8.1.11 Establish critical limits for each CCP (Principle 3)

Critical limits:

Critical limits are defined as criteria that separate acceptability form unacceptability. A critical limit represents the
boundaries that are used to judge whether an operation is producing safe products. Critical limits may be set for
factors such as temperature, time, physical product dimensions, water activity, moisture level, etc. These
parameters, if maintained within boundaries, will confirm the safety of the product.

Sources of information on critical limits include

 Scientific publications/Research data

 Regulatory requirements and guidelines
 Experts (eg. Thermal process authorities, consultants, food scientists, microbiologists, equipment
manufactures, sanitarians, academicians)
 Experimental studies (eg. In house experiments, contract laboratory studies)

Table Examples of critical limits

Hazard CCP Critical limit

Bacterial pathogens Pasteurization 72oC for at least 15 sec.
(Non-sporulating)
Metal fragments Metal detector Metal fragments larger than
0.5mm
Bacterial pathogens Drying over Aw < 0.85 for controlling growth
in dried food products
Excessive nitrite Curing room/brining Max. 200 ppm sodium nitrite in
finished product
Bacterial pathogens Acidification step Max. pH of 4.6 to control C.
botulinum in acidified food
Food allergens Labelling Label that is legible and contains
a listing of correct ingredients
Histamine Receiving Max. of 25 ppm histamine levels
in evaluation of tuna for
histamine

Table Critical limits Vs Operating limits

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 Process Critical limit Operating limit
Acidification pH 4.3

Drying pH 4.6 0.80 Aw

Hot fill 0.84 Aw 85oc

Slicing 80oc 2.5cm

2.0cm
8.1.12 Establish a monitoring system for each CCP (Principle 4)

What will be monitored?

Monitoring may mean measuring characteristics of the product or of the process to determine
compliance with a critical limit.

Examples are:

 Measurement of the time and temperature of a thermal process

 Measurement of cold-storage temperature
 Measurement of pH
 Measurement of Aw

Monitoring may also mean observing whether a control measure at a CCP is being implemented. Eg. i) Visual
examination of sealed cans, and ii) Verification of vendor’s certification of analysis

Establish corrective Actions (Principle 5)

The codex HACCP system and guidelines for its application defines corrective action as “any action to be taken when
the results of monitoring at the CCP indicate a loss of control”

Loss of control is considered as a deviation from a critical limit for a CCP. Deviation procedures are
predetermined and documented. All deviations must be controlled by taking actions to control the non-
compliant product and to correct the cause of non-compliance. Product control includes proper identification,
control and disposition of the affected product. The control and disposition of the affected product and
corrective actions taken must be recorded and filed.

8.1.13 Establish corrective actions (Principle 5)

The codex HACCP system and guidelines for its application defines corrective action as “any action to be taken when
the results of monitoring at the CCP indicate a loss of control”

Loss of control is considered as a deviation from a critical limit for a CCP. Deviation procedures are
predetermined and documented. All deviations must be controlled by taking actions to control the non-
compliant product and to correct the cause of non-compliance. Product control includes proper identification,
control and disposition of the affected product. The control and disposition of the affected product and
corrective actions taken must be recorded and filed.

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 8.1.14 Establish verification procedures (Principle 6)

Verification:

Verification is embodied in HACCP principle 6. The codex guidelines define verification as “the application of
methods, procedures, tests, and other evaluations, in addition to monitoring to determine compliance with the
HACCP plan”, Verification and auditing methods, procedures and tests, including random sampling and analysis,
can be used to determine if the HACCP system is working correctly.

8.1.15 Establish Documentation and Recorded keeping (Principle 7)

Four types of records should be kept as part of the HACCP programme:

 Support documentation for developing the haccp plan

 Records generated by the HACCP system
 Documentation of methods and procedures used
 Records of employee training programme

Chapter 2: Physical, chemical and biological hazards in seafood

8.2.1 Introduction
Hazard analysis is the assessment of all hazards that are associated with the operational steps of the process. Three
hazards under the food safety are

 Physical
 Chemical and
 Biological

Physical hazards: Bones, staples, wood, metal pieces. Other related hazards are short weight, incorrect labels, species
substitution, short counts and decomposition.
Chemical hazards: Heavy metals, pesticides, detergents, disinfectants, diesel oil, machine oils, PSP and DSP .

Biological Hazards:Various bacteria of public health significance, virus and metabolites formed by bacterial
action.

8.2.2 Hazard Analysis

Physical hazards-Analysis
a. stone, sand and mud

 The critical control point is receiving

 This can be controlled by proper verification of the raw material at time of receipt
 Also by washing the raw material thoroughly with water(GMP).

b. Glass pieces:

 The CCP is raw material receiving

 This can be controlled by avoidance of glass from premises of seafood processing plants.
 By providing cover to bulbs, tubes etc.

c. Bones, shell pieces, legs, veins, antennae etc

This is due to improper dressing of the material Rectification – proper dressing of the material and improving the
quality of supervision(GMP)..

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 8.2.2.1 Health Hazards from Pests
Health hazards from pests:

 Effective pest control programme play a key role in overall sanitation of seafood processing factories.
 Insects, rodents and birds can easily enter the processing premises and contaminate the food
 Pest may be divided into 8 groups. Insects and other invertebrates, mammals and birds.
 Relatively few species are encountered in food plant.

Cockroaches Flying insects Rodents Birds

They transfer Flies – good carriers Health risks from They transmit
microorganisms to of bacteria like rodents – plague, microorganisms to
food. Salmonella, Vibrio salmonellosis, foods through their
cholera etc. Jaundice etc. droppings.
Objectionable also
due to their offensive
smell.

Identification and control measures of pests can be effected at following levels

 Proper plant design and operation

 Design and operation
 Destruction within the plant
 Proximity to a refuse dump increases chances of pests
 By proper design pests can be controlled
 Final line of defense is destruction

Best method of control

Cockroaches Flying insects Rodents Birds

Avoid garbage, Repellents
To eliminate habitat, Removal of breeding Avoid chances for and physical
Remove food sources, habitat, Removal of entry, Use of traps barriers
Keeping premises dry, garbage, Fly screens, and rodenticides
Sealing of cracks and Air curtains at entry,
crevices and Application of Electrocutors and
insecticides insecticides

e. Metal pieces f. Filth g. Decomposition of fishery products

From machines, Those engaged in Enzymatic action,

harvest, transport and Oxidative changes,
From outside, processing to be are Due to bacteria
careful.
Metal detectors – final solution

CCP Packaging Raw material,

Receiving- monitoring needed

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 8.2.2.2 Prevention

Prevention

 Checking at the raw material receiving stage

Short weight

 Due to drip loss

CCP: Packaging

Preventive measures

 Use calibrated balance/weight

 Check each days production for weight

Grade size misrepresentations

 Product size has significant impact on its value

Prevention

 Hourly grade checking

 Calibration of weighing scales

Species substitution

 Affect maintenance of product integrity

Control measure

Checking at the time of receipt and at the time of packing and labelling

8.2.3 Chemical hazards-analysis

Presence of a chemical may not always be a hazard. The amount of chemical present is important, Regulatory limits
are set for some of these chemicals.
Chemical hazards belong to 3 groups
i. Naturally occurring chemical hazards

 Scombroid (histamine) poisoning

 Ciguatoxin
 PSP / DSP

ii. Unintentionally added chemical

 Pesticides
 Fungicides
 Toxic elements

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iii. Intentionally added chemical

 Antibiotics -Aquaculture
 Sulphites -Melanosis control
 Nitrites -Preservatives
 Detergents -Cleaning
 Colouring agents -For bright colour
 Antioxidants -prevention of rancidity

Among the Naturally occurring chemical hazards Histamine production is significant in scombroid fishes.

8.2.4 Biological hazards-analysis

Biological Hazards-Analysis

It includes:

 Pathogenic bacteria
 Viruses
 Parasites

These hazards come from Raw materials and food processing steps

 Pathogenic bacteria cause illness in human being either by Infection (by swallowing bacteria) or by Intoxication
( by swallowing free formed toxins). Pathogenic bacteria – mostly absent in fish caught from off shore waters.
Contamination occurs during handling / processing. Time – temperature conditions, if favorable multiplication
is at faster rate.

Bacteria of public health significance

 E.coil
 Faecal streptococcus
 Staphylococcus aureus
 Salmonella
 Shigella
 V. cholerae
 Parahaemolyticus
 V. vulnificus
 L. monocytogenes
 Camphylobacter
 B. cereus
 Cl. botulinum
 Cl. perfringens

Chapter 3: Important biotoxin in seafood

8.3.1 Tetradotoxin

This toxin occurs in different animal species such as pufferfish (Tetraodontidae),a goby, Atelopid frogs Taricha
salamanders, octopus and two Japanese shellfishes. This toxin is mainly found in the liver, ovaries and intestines in
various species of pufferfish. The muscle tissue of the fish is normally free of toxin but there are exceptions. The
occurrence is mainly from ovaries and liver. In octopus, the toxin is present in the posterior salivary venom glands
and will be used for paralyzing the prey. The toxin was isolated as maculotoxin, chemically similar to tetradotoxin
but distinct.

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 The poisoning leads to neurological symptoms 10-45 minutes after ingestion. It acts by preventing nerve
conduction by an internally specific and reversible blockage of the inward movement of sodium ions through the cell
membrane of an activated neuron. Symptoms are tingling sensation in face and extremities, paralysis of respiratory
system and cardiovascular collapse. In fatal cases, death takes place within 6 hours.

Tetrodatoxin(TTX) distribution
1. Fish: pufferfish, Fugu(Tetraodontidae) , Goby (Gobius criniger)

2.Amphibians: Newts, Taricha (Salamandridae) frogs, Atelopus varius,

Atelopus chiriquiensis (Atelopidae)

3. Octopus, Haplaochlae maculosa (Octopoda)

4.Shellfish(Gastropoda) Ivory shell, Babylonia japonica; trumpet shell,

Charonia sauliae

5.Star fish , Astropeeton polyacanthus (Echinodermata)

8.3.2 Ciguatera toxin

This poisoning result from the ingestion of fish that have become toxic by feeding on toxic dinoflagellate.
Gambierdiscus toxicus, which is living around coral reef closely attached to macroalgae. Increased production of
toxic dinoflagellates are seen when reefs are disturbed ( hurricanes, blasting of reefs etc.). The toxin ciguatera
accumulates in fish that feed on the toxic algae. Toxin can be detected in gut, liver and muscle tissue by means of
mouse assay and chromatography. The pharmacological action of ciguatoxin is due to cholinesterase inhibition.
According to Taylor (1988), some fish may be able to clear the toxin from their systems. The clinical picture varies
but onset time is a few hours after ingestion of toxin.
Gastrointestinal and neurological systems are affected due to the toxin. Duration of illness may be 2-3 days but some
may also persist for weeks on even years in severe cases. Death results from circulatory collapse. The most consistently
reported symptoms are

1. Moderate to severe neurological disorders that may persist for days, weeks,or months and/or
2. Moderate to severe gastrointestinal disorders of relatively short duration
3. In some cases, death due to respiratory failure.

Cigutera may be caused by over 400 species of marine fishes, including many that are highly priced for food. Five
species of organisms namely Gambierdiscus toxicus, Prorocentrum concavum, P. mexicanium, Gymnodinum
sanguineum and Gonyaulax polyedra, produced one or more toxic fractions which killed mice within 48 hours.

8.3.3 Paralytic shellfish poinsoning

Intoxication after consumption of shellfish is a syndrome that has been known for centuries, the most common being
paralytic shell fish poisoning, PSP is caused by a group of toxins [saxitoxins and derivatives] produced by dinoflagellates
of the genera Alexandrium., Gymnodinium and Pyrodinium.
Historically, PSP has been associated with the blooming of dinoflagellates [106 cells/liter], which may cause
reddish or a yellowish discoloration of the water.
The dinoflagellate blooms are a function of water temperature, light , salinity, presence of nutrients and other
environmental condition. However, the precise nature of factors eliciting a toxic colour is unknown. Water
temperature must be >5-8oC for blooms to occur. When temperature decrease to below 4oC, the dinoflagellate will
survive as cysts buried in the upper layer of sediments.
Mussels, clams, cockles and scallops that have fed on toxic dinoflagellates retain the toxin for varying periods of
time depending on the shellfish. Some clear the toxin very quickly and are only toxic during the actual bloom, other
retain the toxin for a long time, even years.

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 PSP is a neurological disorder and the symptoms are tingling, burning and numbness of lips and fingertips, ataxia,
drowsiness and incoherent speech. In severe cases, death occurs due to respiratory paralysis. Symptoms develop
within ½ hour to 2 hours after consumption and victims who survive more than 12 hours generally recover.

Toxin is found in their siphon. Gonyaulox tamarensis & Alexandrium catanella the toxin is known as saxitoxin.
Toxin is heat stables not destroyed by cooking. One gram toxic meat can kill 5 people. Estimated lethal dose for
man is 1-4mg, respiratory failure followed by death. Occurrence of this toxin in tropical water is scarce. US FDA
limit is 80mg poison /100g.

8.3.4 Diarrhetic shellfish poinsoning(DSP)

Thousands of cases of gastrointestinal disorders caused by diarrhetic shell fish poisoning (DSP) have been reported in
Europe, Japan and Chile [WHO 1984]. The causative dinoflagellates which produce the toxins are within the genus
Dinophysis and Aurocintrum. These dinoflagellates are widespread which means that this illness could also occur in
other parts of the world. At least seven toxins have been identified including Okadoic acid. Onset of disease is within
1/2 hour to a few hours following consumption of shellfish which have been feeding on toxic algae. Symptoms are
gastrointestinal disorder and victims recover within 3-4 days. No fatalities have ever been observed.
8.3.5 Neurotoxic shellfish poisoning(NSP)

Neurotoxic shellfish poisoning [NSP] has been described in people who consumed bivalves that have been
exposed to red tides of the dinoflagellate [Phycodiscus breve]. The disease has been limited to the Gulf of Mexico
and areas of Florida. Brevetoxins are highly lethal to fish and red tides of this dinoflagellate is also associated
with massive fish kills. The symptoms of NSP resembles PSP except that paralysis does not occur, NSP is seldom
fatal.

8.3.6 Amnesic shellfish poisoning(ASP)

Amnisic shellfishpoisoning [ASP] has only recently been identified [ Todd 1990, Addison and Slenart 1989]. The
intoxication is due to domoic acid, an amino acid produced by the diatom Nitzschia pungens. The first reported
incidence of ASP occurred in the winter of 87/88 in eastern Canada, where, over 150 people were affected and 4
deaths occurred after consumption of cultured blue mussels.

The symptoms of ASP vary greatly from slight nausea and vomiting to loss of equilibrium and central neural deficit
including confusion and memory loss. The short-term memory loss seems to be permanent in surviving victims, thus
the term amnesic shellfish poisoning.

8.3.7 Control of disease caused by biotoxin

The control of marine biotoxins is difficult and disease cannot be entirely prevented. The toxins are all of non-
protein nature and extremely stable (Gill et al.1985). Thus, cooking, smoking, drying, salting does not destroy them,
and one cannot tell from the appearance of fish or shellfish flesh whether it is toxic or not.
The major preventive measure is inspection and sampling from fishing areas and shellfish beds and analysis for
toxins. The mouse bioassay is often used for this purpose and confirmatory HPLC is done if death occurs after 15 min.
To be effective, the monitoring required reliable sampling plans and efficient means of detection of the
toxins. Reliable chemical methods for detection of all toxins are at present available. Also the toxicity can vary within
a growing location of shellfish according to geography of water currents and tidal activity.

Toxin Tolerance Method of analysis

Ciguatera Control not possible No reliable method
PSP 8.0 ug/100g Mouse bioassay, HPLC
DSP 0.60 ug/100g Mouse bioassay HPLC
NSP Any detectable level/100 g is unsafe Mouse bioassay no chemical method
ASP 20 ug/g demoic acid HPLC

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 8.3.8 Toxic metals

Toxic metals like Cd, Hg, Se, Pb etc. above normal level affect the safety and marketability of foods. Many countries
have standards. Older ones have higher levels than younger ones of the same species. Presence of cadmium in
cephalopods exported from India is reported. Control by minimizing pollution & environmental monitoring

Organochlorine pesticides & polychlorinated biphenyls

 OCP & PCBs now common is all environments

Important compounds of concern are

 DDT and its derivatives

 Aldrin and Dieldrin
 Benzene hexachloride
 Polychlorinated biphenyl

It pose special problems in the environment because they are

 Extremely persistent
 They are lipophilic is nature
 Filter feeders like oysters, mussels & clams accumulate these compounds

Permitted levels in seafood

Compounds Max. residue limit (mg/kg)

DDT and its 5

derivatives

Dieldrin 3

PCB 2

Mercury 0.5

Cadmium 3-Crustacean; 4- mollusc

Lead 1-5-crustacean; 1-7 bivalves

Control

 Prevention of contamination and Environmental monitoring

8.3.9 Antibiotic Residues

Widely used in shrimp culture to prevent disease. Usual practice to withdraw the drug in advance before harvest to
enable the animal to clear off the residue. Sometime cultural shrimp contains high quantities. In-1992 – 1994 Japan
rejected many shrimp consignments from Thailand. Japanese food sanitation act prohibits antibiotic residues in
food. From 1994 November onwards US FDA insists testing antibiotic residues in fish

Control:

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  Avoid misuse of antibiotics
 Give sufficient time to clear – off the residue

Food additives:
Food additives including colours are used in various fish & fishery products ( Council directive 95/2 EC on Food
Additives )

Chapter 4: Histamine

8.4.1 Biogenic Amines

Histamine poisoning is a chemical intoxication following the ingestion of foods that contain high levels of
histamine. Historically, this poisoning was called scombroid fish poisoning because of the frequent association with
scombroid fishes including tuna and mackerel. The most common symptoms are cutaneous such as facial flushing,
utricaria, edema, but also the gastrointestinal tract may be affected ( nausea, vomiting, diarrhea) as well as
neurological involvement ( headache, tingling, burning sensation in the mouth).
Histamine is formed in the fish post mortem by biological decarboxylation of the amino acid histidine.

The histamine-producing bacteria are certain enterobacteriaceae, some Vibrio sp. A few Clostridium and
lactobacillus sp. The most potent histamine producers are Morganella morgani, Klebsiella pneumoniae and Hafinia
alviei ( Etratten and Taylor, 1991). These bacteria can be found on most fish, probably as a result of post-harvest
contamination they grow well at 10o C but at 5o C growth is greatly retarded.

The principal histamine producing bacteria M.morganii grow best at neutral pH, but they can grow in the pH
range 4.7-8.1.The organism is not very resistant to NaCl, but at otherwise optimal conditions growth can take place
in upto 5% NaCl. It should be emphasized that at once the histamine has been produced in the fish, the risk of
provoking disease is very high. Histamine is very resistant to heat, so even if the fish are cooked, canned or
otherwise heat-treated before consumption, the histamine is not destroyed.

The human body will tolerate a certain amount of histamine without any reaction ingested histamine will be
detoxified in the intestinal tract by at least 2 enzymes the diamine oxidase (DAO) and histamine N-methyl transferase
(HMT) (Taylor, 1986).
Their protective mechanism can be eliminated if intake of histamine and / or other biogenic amines is very higher
if the enzymes are chocked by other compounds as shown in figure. Other biogenic amines such as cadaverine and
putriscine which are known to occur in spoiled fish may therefore act as potentiators of histamine toxicity. Presumably
inhibition of intestinal histamine catabolism will result in greater transport of histamine across cellular membrane and
into the blood circulation.
8.4.2 Control of Disease caused by biogenic amines
Low temperature storage and holding of fish at all times is the most effective preventive measure. All studies
seems to agree that storage at Oo C or very near to 0o C limits histamine formation in fish to negligible levels. Several
countries have adopted regulations governing the maximum allowable levels of histamine in fishes. e.g.

Regarding limit for histamine in fish

Hazard action Defect action Maximum

allowable allowable allowable
level mg/100g level mg/100g level mg/100g

USA (FDA) 50 10-20 --

EEC ----- 10 20

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Toxin Where/when Animals/organisms
produced involved
Tetradotoxin Fish Puffer fish, mostly
ovaries, liver,
intestine
Cignatera Marine algae Tropical/subtropical
fish
PSP Marine algae Filter feeding
shellfish
DSP Marine algae Filter feeding
shellfish

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