P.O.

Box 1390, Skulagata 4

120 Reykjavik, Iceland

Final Project 2005

GUIDELINES FOR HANDLING AND PRESERVATION

OF FRESH FISH FOR FURTHER PROCESSING IN VIETNAM
Nguyen Huy Quang
Quality Assurance Department
Seafood Export and Quality Improvement Program
Vietnam

[email protected]
Supervisors
Prof. Hjrleifur Einarsson, [email protected]
Ms. Arnheiur Eyrsdttir, [email protected]
University of Akureyri
ABSTRACT
Fish from catching has an important role in international fisheries as well as in
developing countries like Vietnam. Therefore maintaining good quality in fish raw
material is necessary. This project focuses on how to handle and preserve the fish
especially during the process from catching the fish at sea to landing and transporting
the fish to the processing plant. This project establishes guidelines for these activities.
In addition some experiments were carried out to determine the insulation ability of
different types of fish boxes used for storing fish and to validate the guidelines by
evaluating the fish quality during ice storage in the worst and best scenario cases.
Based on data collected in Vietnam as well as fish preservation techniques in Iceland,
problems in the handling and preservation process in Vietnam are pointed out and
solutions presented. Choosing the appropriate fish containers like boxes or tubs is
considered one significant factor contributing to fish freshness and quality. The
Splast insulation plastic boxes or tubs are very suitable containers, which can
possibly be used in the Vietnamese fisheries industry in the near future.

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TABLE OF CONTENTS
1

INTRODUCTION .................................................................................................................................... 5

LITERATURE REVIEW ....................................................................................................................... 7

2.1
2.1.1
2.1.2
2.1.3
2.1.4
2.2
2.3
2.3.1
2.3.2
2.3.3

REASONS FOR SPOILAGE OF FISH ................................................................................................................7

Autolysis ........................................................................................................................................................7
Bacteria8
Rancidity........................................................................................................................................................9
Mechanical damage ....................................................................................................................................9
FISH RAW MATERIAL HANDLING AND PRESERVATION ..........................................................................10
A NALYSIS METHODS FOR QUALITY EVALUATION ..................................................................................12
Sensory method ..........................................................................................................................................12
Microbiological methods..........................................................................................................................12
Chemical methods......................................................................................................................................13

3
DESCRIPTION OF THE PRESENT SITUATION AND PROCEDURES IN
VIETNAMESE AND ICELANDIC FISHERIES ...................................................................................... 13
3.1
3.1.1
3.1.2
3.1.3
3.1.4
3.1.5
3.1.6
3.1.7
3.1.8
3.1.9
3.2
3.2.1
3.2.2
3.2.3
3.2.4
3.2.5
3.2.6
3.2.7
3.2.8
3.2.9

PRESENT SITUATION OF FISH HANDLING IN VIETNAM...........................................................................13

Catching method........................................................................................................................................17
Sorting 17
Primary washing........................................................................................................................................18
Icing in boxes..............................................................................................................................................18
Trading and transporting at sea..............................................................................................................19
Unloading....................................................................................................................................................19
Trading and transporting on land...........................................................................................................19
Re-icing and pre-processing in the trading establishment or middlemen .......................................19
Reception at the factory............................................................................................................................20
PRESENT SITUATION OF FISH HANDLING IN ICELAND............................................................................20
Catching method........................................................................................................................................21
Boarding ......................................................................................................................................................21
Holding 21
Sorting 22
Bleeding and gutting .................................................................................................................................22
Washing .......................................................................................................................................................22
Chilling 22
Storing 22
Landing to plant or auction market........................................................................................................22

MATERIALS AND METHODS ........................................................................................................ 23

4.1
4.2
4.2.1
4.2.2

EXPERIMENT FOR DETERM INING THE INSULATION ABILITY.................................................................23

EXPERIMENT FOR THE QUALITY CHANGE IN FISH ..................................................................................24
Fish preparation ........................................................................................................................................24
Measurements.............................................................................................................................................24

RES ULTS AND DISCUSSION .......................................................................................................... 25

5.1
5.2
5.2.1
5.2.2
5.2.3

ICE MELTING EXPERIMENT ........................................................................................................................25

DIFFERENCE OF FISH QUALITY IN THE WORST AND BEST SCENARIO CASE .........................................27
Sensory quality...........................................................................................................................................27
Chemical analysis......................................................................................................................................29
Microbial analysis .....................................................................................................................................30

RECOMMENDATIONS FOR IMPROVEMENTS IN VIETNAMESE FISHERIES ....... 32

CONCLUSIONS ..................................................................................................................................... 34

ACKNOWLEDGEMENTS .............................................................................................................................. 36

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LIST OF REFERENCES .................................................................................................................................. 37

APPENDIX 1: ANALYSIS METHOD (SENSORY, MICROBIOLOGY AND CHEMICAL) .... 39
1. SENSORY METHOD ..............................................................................................................................................39
1.1
Material.......................................................................................................................................................39
1.2
Method 39
2
M ICROBIOLOGICAL METHOD ....................................................................................................................41
2.1
Material.......................................................................................................................................................41
2.2
Method:41
3
CHEMICAL METHOD ...................................................................................................................................41
3.1
Material.......................................................................................................................................................41
3.2
Method:42
APPENDIX 2 - SENSORY TESTING RESULTS ..................................................................................... 43
APPENDIX 3 - CHEMICAL TESTING RES ULTS ................................................................................. 45
APPENDIX 4 - MICROBIOLOGY TESTING RESULTS ..................................................................... 47
APPENDIX 5 - RESULTS OF ICE MELTING AMOUNT BY TIME FOR SOME TYPES OF
BOX/TUB .............................................................................................................................................................. 55

LIST OF FIGURES
Figure 1: Fish production in Vietnam (FAO 2003)...................................................................................... 6
Figure 2: Change in micro-organism and enzyme growth by temperature (Huss 1994). ................. 9
Figure 3: Four ways ............................................................................................................................................ 15
Figure 4: Flow chart for fish in Vietnam. ..................................................................................................... 16
Figure 5: Flow chart for handling of fish and processing in a typical trawler in Iceland catching
mainly cod and haddock. ........................................................................................................................ 20
Figure 6: Catch of trawler by fishing gear in Iceland 2005 (Statistic Iceland 2005). ....................... 21
Figure 7: The types of box/tub used for experiment (a) Splast tub 70 l, (b) Splast cooler 65 l,
(c) Vietnamese-like box (VN box)......................................................................................................... 23
Figure 8: Weight of ice per container by time for the five different containers. ............................... 25
Figure 9: Melting rate of ice in box used in the experiment. ................................................................... 26
Figure 10: Sensory score (QIM) of fish stored in a VN box and Splast box. ................................... 28
Figure 11: TVB-N contents of fish in ice stored in VN and Splast containers. ............................... 29
Figure 12: Total bacteria counts (PCA, 30C) in the VN box and the Splast box.......................... 30
Figure 13: Total bacteria counts (IA, 22C) in the VN box and the Splast box.............................. 31
Figure 14: Black colony counts (IA, 22C) in the VN box and the Splast box. ............................... 31

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LIST OF TABLES
Table 1: The relative change in abundance of different groups of bacteria in cod stored in ice
(Hobbs 1982). ............................................................................................................................................... 8
Table 2: Shelf life of cod stored at 0C and predicted shelf life at 5, 10 and 15C (adapted from
Huss 1994)................................................................................................................................................... 10
Table 3: Physical characteristics of ice utilised in chilling fish (Huss 1994). ...................................... 11
Table 4: Time and temperature parameters in each stage from catching to the fish processing
plant (Tam et al. 2004)............................................................................................................................. 14
Table 5: The identified risk as low, medium and high in each handling stage of raw material in
the flow chain. ............................................................................................................................................ 17
Table 6: Sampling schedule for evaluation. ................................................................................................. 24

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INTRODUCTION

Vietnam has great potential for fish exploitation with a coastline over 3,260 km long.
The inland water area is about 226,000 km2 and the Exclusive Economic Zone (EEZ)
is over 1 million km2 , three times the mainland area. Marine capture is divided into
two fishing seasons with different characteristics, the south season (from March to
September) and the north season (from October to February). There are more than
2000 fish species in Vietnamese marine waters, of which about 130 are of economic
value. These are species like tuna, mackerel, swordfish, mahi mahi, scads, herring,
sardine and demersal fish like sole, hair tail, pomfret, sea bream, grouper, sea perch
and snapper. Coastal fisheries are characterised by high species diversity and small
short- lived species. The resources have high potential for recovery and can sustain
high levels of harvest. Besides marine fish, there are over 1600 species of crustaceans
and about 2500 species of molluscs where squids and octopus are of significant
economic value. There are some fresh water fishes with high economic value like
catfish, snakehead, perch, tilapia and eel. There is also potential for fish aquaculture
in Vietnam with its long coastline, many lagoons, straits and bays, estuaries, canals
and thousands of small and big islands. In the inland area, many rivers, canals,
irrigation and hydroelectric reservoirs have created a water surface area of about
1,700,000 ha. Fish production in Vietnam is developing quite fast (Figure 1) reaching
3.2 million tons in 2004. Out of this total, capture fisheries contributed 1.7 million
tons, mainly from coastal fisheries (1.1 million tons). Although the contribution of
capture fisheries is high in terms of volume the bulk of the catch is made up by low
value fish, except for cephalopod and tuna. Fish aquaculture product yield was 1.5
million tons in 2004 with the main species being catfish (basa and tra fish) and blacktiger shrimp (Ministry of Fisheries 2005).
Today in Vietnam the consumption rate of fish for food is about 50% of the total
protein food. The people prefer seafood products more and more. Fish consumption
per person is still rather low at 8 kg/year. Therefore this amount needs to increase.
Fish products are exported to many countries in the world, in which the main markets
are the EU, USA and Japan. The total exporting value has been increasing for many
years. The total fish products export value in 2004 was USD 2.35 billion for products
mainly from finfish, shrimp and squid. The increase of capture fisheries is declining
as stocks are becoming fully exploited. Therefore, maintaining the quality of fish raw
material is more and more important. If the fish quality can be maintained the value
from each trip for catching at sea can continue to increase. The fish product volume
for export and domestic consumption can increase if the raw material used for
processing is of higher quality (Ministry of Fisheries 2005).
Quality of fish raw material plays an important role for the quality of the end-product.
Once the fish raw material freshness and nutrition value is lost, it can not be
recovered in the processing stages. Products that are processed from low quality raw
material are not always a safety risk, but the quality (nutrition value) and shelf life is
significantly decreased.

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3000

Yield (ton x 1000)

2500
2000

1500
1000

500
0
1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003
Year
Total

Catching

Aquaculture

Figure 1: Fish production in Vietnam (FAO 2003).

In Vietnam, maintaining fish raw material quality is still a challenge. The time from
catching to reception at a processing plant can be prolonged while the temperature of
the raw material usually is not low enough to prevent spoilage. Some exporters have
experienced product rejection due to quality problems e.g. microbiological criteria or
extraneous matter. So the Vietnamese fishing sector is facing at least two serious
problems, one is stagnation in catching and the second is deterioration in raw material
quality. In light of the decrease in catch rates, quality improvements become
especially important. There are three main reasons for quality deterioration and
spoilage: autolysis, bacterial activity and rancidity (Huss 1995) and in some cases
physical damages (mechanical stress, direct sunlight etc) can lower the quality
considerably. The quality deterioration can start right away during fishing and it
continues all the way to the final user.
The main objective of this project is to introduce new and validated guidelines for
handling and preservation of fish in Vietnam in order to improve the quality of the
raw material. Some studies on quality changes in fish under best and worst case
scenarios were carried out to validate these guidelines. Some tests for the suitability
of different plastic boxes or tubs for storing fish, were also carried out to examine
these best and worst case scenarios in terms of retaining quality.

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LITERATURE REVIEW

2.1

Reasons for spoilage of fish

Maintaining good quality of fish raw material for processing is very important.
Therefore, the reasons for quality deterioration leading to spoilage need to be
determined carefully. Just after death, fish can be soft for a few hours but then it
becomes stiff. This phenomenon is called rigor mortis. The fish stays in the rigor
mortis condition for a while, but then its flesh muscles become relaxed again. At that
time the fish quality starts to decrease. The quality changes can easily be noticed and
consist of changes in colour, odour or smell, taste, appearance and texture and are
therefore called sensory changes. One of the differences between fish appearance
before and after rigor mortis is that the fish muscle is more elastic before rigor mortis.
The time of pre-rigor mortis and rigor mortis varies according to species. It also
depends on many things like temperature, handling, size and physical condition of the
fish.. Generally, it is preferred to extend the time before and during rigor mortis.
There are some reasons for deterioration of quality and spoilage; they are autolysis,
bacteria spoilage, rancidity and mechanical damage (Huss 1994). Lowering the
temperature by icing not only slows down the rigor mortis process, but also reduces
the spoilage rate. Therefore maintaining low temperature during the handling and
preservation process is very important.
2.1.1

Autolysis

The autolysis process relates to enzyme activities in fish (autolysis means selfdigestion). Commonly the spoilage due to autolysis occurs first and is followed by
spoilage due to bacteria and rancidity but sometimes they overlap (Gram and Huss
1996). Unlike most fish, autolysis occurs very quickly in some shellfish like lobster
and shrimp (Hobbs 1982). When the fish dies adenosine-triphosphate (ATP), which is
the energy-rich organic compound in its muscle, will mostly be synthesised from
glycogen, but also from creatine-phosphate (for finfish) and from arginine-phosphate
(for cephalopods) under anaerobic conditions. The glycolysis (glycogen reduction
process) still occurs continuously to create the end product of lactic acid. Because the
end product of this process is lactic acid, the pH of the muscle will decrease. The
ATP concentration gradually decreases and when it goes below 1 mol/g in the
muscle tissue the enzyme ATP-ase is activated. This leads to the stiffing of the
muscle which will be constant (rigor mortis). The ATP is gradually degraded during
time to some degraded products e.g. adenosine diphosphat, adenosine monophosphat,
inosin monophosphat, inosin and hypoxanthin. Hypoxanthin is considered to cause
the off- flavour in spoiled fish. When the fish raw material is handled carelessly cells
may be broken, which leads to the release of autolytic enzymes and this leads to the
production of some spoilage substances. These substances create a very good
environment
for
micro-organisms.
Cathepsin,
chymotrypsin,
trypsin,
cacboxypeptidase, calpain, collagenase and TMAO- demethylase are all autolytic
enzymes. Therefore, in order to maintain fish quality, enzyme activities should be
prevented. Using low temperature is the most frequently used measure to limit
enzyme activities (Huss 1994).

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2.1.2

Bacteria

Bacteria are capable of causing spoilage because of two important characteristics.

First they are psychotropic and thus multiply at refrigeration temperatures. Secondly
they attach various substances in the fish tissue to produce compounds associated
with off- flavours and off-odours. When the fish is alive the bacteria are found on the
gill and skin and in the intestines but can not attack the fish muscle. But when the fish
dies the bacteria can penetrate into the flesh muscle of the fish. When fish is
preserved by icing the rate of bacterial penetration into the flesh muscle is much
slower. Fish spoilage occurs when the enzyme of bacteria diffuses into the flesh
muscle and the nutrition substances from the flesh muscle diffuse to the outside.
Spoilage will happen more rapidly for fish species with a thin skin layer. The number
of bacteria in fish caught in temperate waters can develop even when in ice but the
bacteria caught in tropical water grow slowly for one or two weeks in icing
preservation (Gram and Huss 1996).
There are many bacteria species present in spoiling fish but there are only certain
types that are considered to cause spoilage. The bacteria use their enzyme to change
fish odour and flavour to sour, gassy, fruity and finally ammonia and faecal odour
appear. Bacteria can still develop during icing as indicated by Hobbs (1982) (Table 1).
Table 1: The relative change in abundance of different groups of bacteria in cod
stored in ice (Hobbs 1982).
Bacteria
Pseudomonas
Achromobacter
Flavobacterium
Coryneform
Micrococcus
Total

0 day (%)

5 days (%)

10 days (%)

15 days (%)

14
33
4
41
8
100%

17
49
0
33
1
100%

50
38
0
12
0
100%

82
14
2
2
0
100%

Not all the growing bacteria are involved in the spoilage process. There are just a few
bacteria species that become predominant and are mainly responsible for spoilage.
For example in gutted cod, chilled by ice the specific spoilage organism (SSO) is
Shewanella putrifaciens and in packaged cod fillet it is Photobacterium phosphoreum
(Connell 1995). If the fish is preserved by icing or in lack of air the amount of
Pseudomonas and Shewanella putrifaciens bacteria is not very high but
Photobacterium phosphoreum bacteria becomes quite high. After a certain time in ice
in aerobic conditions the Pseudomonas and Shewanella putrifaciens bacteria will
become the predominant bacteria. In general in low temperature (0-5C), Shewanella
putrefaciens, Photobacterium phosphoreum, Aeromonas spp., and Pseudomonas spp.
cause spoilage but in higher temperature (15-30C) other species like Vibrionaceae,
Enterobacteriaceae and the positive Gram bacteria cause spoilage (Gram and Huss
1996). The bacteria produce a high amount of volatile compounds. These are
trimethylamine, volatile sulfur compounds, aldehydes, ketones, esters, hypoxanthine
as well as other low molecular weight compounds. The bacteria S. putrefaciens and
some Vibrionaceae produce H2 S but Pseudomonas and Photobacterium

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phosphoreum do not produce significant amounts of H2 S. The volatile sulphurcompounds have a very bad odour so even minimal quantities are considered to affect
quality. The low temperature is very important in preservation of raw material.
Especially in the range of 0-25C the temperature strongly affects the bacteria
activity (Figure 2). At 0C the bacteria grow very slowly. The typical spoilage
bacteria like Shewanella putrefaciens develop 10 times less in comparison with
growing at the optimal temperature. Raising the keeping temperature thus increases
the spoilage rate rapidly. Therefore it is important to decrease the temperature to 0C
as soon as possible after catching. For fish in the tropical water area where the
ambient temperature is around 25 30C the rate of spoilage can be 25 times higher
than when kept at 0C (Huss 1994).

Figure 2: Change in micro-organism and enzyme growth by temperature (Huss 1994).

2.1.3

Rancidity

Fat oxidation usually occurs after autolysis and bacterial spoilage. The lipid
concentration in fish can contribute to the spoilage process in fish. The fats in fish are
mainly unsaturated fatty acids that are easily oxidised by oxygen from the atmosphere.
High temperature or exposure to light can increase the oxidation rate. For fatty fish
preserved in ice, spoilage due to rancidity is mainly caused by oxidation. This
produces a bad and unpleasant odour as well as a rancid taste (Hobbs 1982). Fat fish
species like herring, mackerel, and salmon are mostly affected by rancidity. The lean
fish fat content is about 0.1-0.9% and the fat fish fat content is higher than 0.9%
(Love 1982)
2.1.4

Mechanical damage

If the fish is broken by harsh handling, it will be subject to mechanical or physical

damage and become bruised and defected in outside appearance. But it is more
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important that some small cells will break leaving the enzymes free to react with
other substances. Mechanical damage gives good conditions for some enzymatic
activities. Fish kept in thick layers in a box with ice can cause high pressure between
the ice and fish causing cells to break. All careless handling of fish raw material can
result in bruised fish. This also opens channels for the micro-organisms to enter the
fish flesh and enables quicker spoilage of the fish (Huss 1995). In general, in order to
maintain the fish raw material quality after catching, some measures for handling and
preservation are needed to prevent all the quality change processes mentioned above.
2.2

Fish raw material handling and preservation

Immediately after catching the fish start to spoil in one way on the other. However the
rate of spoilage is different depending on ambient conditions, fishing technology,
fishing equipment, species of fish, catching season and handling and preservation
activities (Hobbs 1982). Using low temperature with ice is a popular method for fresh
fish preservation. The chilling temperature of nearly 0C can maintain freshness
quality for a long time. When the temperature decreases the bacterial growth is
slower, the reaction rate of enzymes is also decreased and the rigor mortis time can be
extended. If the shelf life of some fish products stored at 0C is known, the shelf life
at different temperatures can be calculated by a certain formula e.g. if the fish can
maintain quality for six days at 0C the shelf life at 5C will be 2.7 days or if another
fish can maintain quality for 10 days at 0C the shelf life at 15C will be only 1.6 day
(Table 2).
Table 2: Shelf life of cod stored at 0C and predicted shelf life at 5, 10 and 15C
(adapted from Huss 1994).
Shelf life at 0C of stored
temperature (days)
6
10
14
18

Shelf life at different temperature (days)

5C

10C

15C

2,7
4,4
6,2
8

1,5
2,5
3,5
4,5

1
1,6
2,2
2,9

Fish chilling should be carried out quickly and the fish raw material should not be
exposed to sunshine or wind. Sunshine and wind can speed up not only autolytic and
bacterial spoilage but also the oxidation process especially in fatty fish species. Fish
handling and preservation can be carried out on board of the fishing vessel or on land.
The first pre-processing stages for whole fish include some stages e.g. bleeding,
gutting, icing and freezing. Some fish species can be bled and gutted on board, but
this work can take much time and some fish species are only primarily washed and
put into boxes or tubs with ice and stored in the hold of the vessel (Kelman 1992)
There are a lot of enzymes in the fish intestine which can be activated strongly when
the fish dies. Fish intestines contain many enzymes catalysing autolysis and spoilage
in fish. Fish intestines also contain many undesirable micro-organisms which can
contaminate the fish flesh. Removing intestine can eliminate these undesirable
enzymes and micro-organisms. Thus it is preferred to bleed and gut the fish, before

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chilling and storing. However, after gutting the inside of the belly area is exposed to
air, which can lead to oxidation and discolouration of the fish. Therefore, some fat
fish species are not always gutted before chilling especially the small sized fish, as
gutting them takes too long time. For the lean fish species, gutting is usually carried
out because this can retain the quality for extended time periods. The chilled sea
water (CSW) that includes ice and seawater can chill the fish raw material very fast.
However if the fish is kept in water for a long time some colour pigments from the
skin as well as some soluble and nutrition substances can be released and loaded into
the environment. Using CSW can also create sensory changes in the fish e.g. higher
salt content after chilling and storing. The chilled water (CW) is also often used for
chilling fish and this does not affect to the fish salt degree (Huss 1994).
Different types of ice can be used for chilling fish like liquid ice, flake- ice, tube ice,
and block ice (Table 3). Block ice should be grinded before use. Liquid ice has the
highest cooling rate, the second is flake ice but grinded block ice is the slowest.
Liquid ice has uniform particle size and large surface squares which means better heat
transfer. Following Huss (1994) the crushed block ice and the tube ice is suitable for
the chilled sea water (CSW) system. The rate of chilling is important. For some big
fish species (e.g. ocean tuna) chilling is carried out by gutting and putting ice into the
belly of fish to increase the chilling rate.
Table 3: Physical characteristics of ice utilised in chilling fish (Huss 1994).
Types

Approximate
Dimensions

Specific volume
(m3 /t)

Specific weight
(t/m3 )

Flake
Plate
Tube
Block
Crushed block

10/20 - 2/3 mm
30/50 - 8/15 mm
50(D)- 10/12 mm
Variable
Variable

2.2 -2.3
1.7 - 1.8
1.6 - 2.0
1.08
1.4 - 1.5

0.45-0.43
0.59-0.55
0.62-0.5
0.92
0.71 -0.66

A common way to chill the fish is to arrange it with ice in a fish box. There are some
specific requirements for these boxes. Research from a long time ago shows that the
fish box should be made by material that is easy to clean; therefore wooden boxes
should not be used as they are porous and with a rough surface (Jrgensen 1965).
Some experiments show that the plastic container is better than aluminium, wood or
wicker basket containers to maintain low temperature and retain the fish quality
stored in these containers (Vyncke 1965). In general fishermen like using larger
containers or tubs because there are fe wer units to handle, saving time for unloading
but the disadvantage in terms of fish quality is the high pressure on the fish in the
bottom of the tub. However the box size must also depend on whether they are
transported by hand or machine force. Tubs with iced fish should have good drainage
to discharge water from the melting ice (Valdimarsson 1992). For a box or a tub
containing fish the thermal insulation is essential to minimise ice consumption and to
keep inside temperature more independent of outside temperature. A prototype from
the 1970s was lined with glass fibre on a frame of iron. Prototypes lined with
aluminium plates were also made. Both prototypes became rather clumsy and there
were heat leakages because of metal contact from the inner to the outer lining.
Polyurethane can be injected in between a double wall for insulation and to give

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increased strength, especially if the polyurethane can be made to adhere to the linings
(Ryrvik 1982). Some requirements for fish boxes are recognised. For example, the
box should have good insulating effects, but the difference between polystyrene
boxes, fibre board boxes and wooden boxes is not well distinguished (Wignall 1982).
However, one of the main requirements for a fish box, tub or container is how to
maintain fish freshness quality and extend the shelf life of fish. Shelf life of fish
relates to handling and preservation methods and some other factors, even to the
fishing season (Kolakowska 1992). In general the fish raw material is stored in the
fish container with ice until reception at the processing factory. To evaluate the raw
material quality like freshness or shelf life, various methods are used. They are
outlined in the following section.
2.3

Analysis methods for quality evaluation

The methods of assessing freshness can be divided to two groups: sensory methods
and non-sensory methods, where non-sensory methods include microbiological,
chemical and physical analysis. Sensory assessment is a direct measure but the nonsensory methods are indirect measurements. They should be used in combination
(Howgate 1982). The disadvantage of the sensory method is that it is subjective
depending on the person who evaluates and people (panellists) have to be trained for
fish sensory evaluation. The non-sensory methods are biological, chemical, physical.
Their disadvantage is complexity because they require laboratory equipment
(Jonsdottir 1992)
2.3.1

Sensory method

Sensory evaluation is a systematic assessment of the odour, flavour, appearance and

texture of food. The Quality Index Method (QIM) is a seafood freshness quality
control system that was developed by European fisheries research institutes. It is
considered to be a rapid and reliable method for assessing freshness. (Martinsdottir et
al. 2001) QIM is based on the significant sensory parameters for raw fish when using
many parameters and a score system from 0 to 3 defect points (see QIM form in
Appendix 1, Table 7). QIM is a practical rating system where the defect points are
recorded. The sum of scores for all the characteristics is the overall sensory score.
QIM gives scores of zero for very fresh fish, while increasingly larger totals result as
the fish deteriorates. The description of evaluation of each parameter is written in a
guideline. When the score is 18 or more the fish is considered spoilage.
2.3.2

Microbiological methods

There are a lot of microbiological methods to determine fish bacteria e.g. plate count,
direct microscopic count, ATP measuring, but the plate count is a traditional and
common method with some different media like plate count agar or iron agar. Some
spoilage bacteria can produce H2 S (e.g. Shewanella putrefaciens) and reduce TMAO.
The iron agar medium can be used in order to isolate spoilage bacteria that produce
H2 S and form black colonies on the agar media. Black and white colonies are
observed and counted respectively. The black ones are referred to as spoilage bacteria,
while the totals (black + white) are referred to as the total count. The pour plate

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method is often used with plate count agar, which is a common method to determine
the total content of bacteria in seafood. The iron agar method can sometimes detect
higher bacteria amounts than plate count agar (Gram 1992).
2.3.3

Chemical methods

Chemical methods to measure freshness quality have been considered to be objective

methods and therefore superior (less variable) to methods involving sensory
evaluation. During post mortem storage microbiological spoilage causes the
formation of volatile bases, which can be determined to measure indirectly the
freshness quality of such seafood. There are a few substances that are usually
determined to evaluate fish raw material freshness, e.g. total volatile basic nitrogen
(TVB-N), trimethylamine (TMA), ammonia, biogenic amines, ethanol and indol. The
TVB-N remains constant for the first days of storage or increases slowly but it rises
fast later in the spoilage process. Therefore TVB-N is a very good indicator of
spoilage in fish (Oehlenschlager 1992). For some types of ground fish species like
Atlant ic cod (Gadus morhua), European hake (Merluccius merluccius), and haddock
(Meranogrammus aeglefinus), the TVB-N determination is not as good to detect the
early stages of deterioration in freshness quality like the TMA measurement, but it
can be used for measuring later stages of deterioration (Botta 1995).

DESCRIPTION OF THE PRESENT SITUATION AND PROCEDURES IN

VIETNAMESE AND ICELANDIC FISHERIES

Handling of raw material can roughly be divided into two categories: the artisanal
type and the industrial type. In general, in the modern industry system the activities
are automated, using little human force. But in Vietnam, the handling is mainly
artisanal and uses mainly human force. The gutting stage is not carried out. After
landing, the fish raw material still has to go through many stages before entering the
receiving area of the factory. This takes a long time and the temperature can easily
fluctuate during the process. In order to keep the ice melting rate slow, using
insulation boxes for storing and transportation of fish with ice is important especially
in tropical areas. In Vietnam very little attention has been paid to the effect storage
boxes that influence the quality of raw material. In Vietnam there are some
experiences in proper handling and processing from catching to factory reception.
This chapter provides some data on temperature and time during the flow chain in
processing fish raw material as well as information related the handling process. The
knowledge which has been gained in Iceland for the last six months is also applied to
point out the main problems in handling and preservation as well as the way to
improve the present situation in Vietnam. It is of prime importance to analyse the
operating procedures along the whole chain from catch to the consumer and to
suggest changes in order to improve the quality of Vietnamese fish.
3.1

Present situation of fish handling in Vietnam

Today in Vietnam, the exploitation fisheries situation is confusing. The off- shore
vessels are not working effectively and fish catch is not high. The catching process at

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sea usually takes a long time, so if the fish is not handled properly, it loses a lot of
value, and the economic gain for the fishermen can be very low. At the fishing port or
fishing market, the middlemen1 sometimes press the price to the fishermen down.
After buying raw material from the fishermen, the middlemen will sell it to trading
establishments 2 , the more trading time the more quality decrease. The factory could
still have to buy this raw material for a high price, especially at times of low supply.
A solution to this may be the formation of an auction market, where the fishermen
could sell at a price according to their fish value and quality. In order to retain high
value, the fishermen and all the other people involved have to know about fish raw
material quality and how to maintain the freshness as high as possible.
In practice the fish can go through a lot of middle stages before entering the
processing factory. This increases the holding time which results in quality loss. If
other conditions are also undesirable such as temperature, hygiene or methods of
handling and transportation the fish quality may decrease much more. The
temperature and time parameters are very different in the flow chain depending on the
type of catching vessel. There are three main types of such vessels in Vietnam: big,
medium and small size vessels. The big vessels usually operate on fishing trips at sea
lasting up to 10 days. The medium vessels stay at sea for about three to seven days
and the small vessels fishing trips last for less than three days. For all of three types
of vessels there are four ways to transfer the raw material to the factory (Figure 3).
The minimum way (way 1) from catching to processing plant takes 4 hours (6)
(Table 4) i.e. when the fishermen sell their fish directly to the factory. For the big
vessels the time can be as long as 270 hours (1 + 2 +3) in the worst case, i.e. a lot
of middle stages before the fish arrives at the factory. The time may be shortened by
transferring the catch at sea to other vessels coming back earlier.
Table 4: Time and temperature parameters in each stage from catching to the fish
processing plant (Tam et al. 2004).
Time () and
Temperature (t)*
1
t1
2
t2
3
t3
4
t4
5
t5
6
t6
* See Fig. 3

Small size
4- 10 hours
15 -30 0 C
1-3 hours
15 -30 0 C
1-2 hours
0-60 C
1-3 hours
10-25 0 C
4- 10 hours
0-60 C
4- 10 hours
15 -30 0 C

Type of catching vessel

Medium size
36-168 hours
0-150 C
3-24 hours
15 -30 0 C
1-6 hours
0-60 C
3-24 hours
10-250 C
36-144 hours
0-60 C
36-144 hours
0-150 C

Big size
168-240 hours
5-150 C
3-24 hours
15 -30 0 C
1-6 hours
0-60 C
3-24 hours
10-250 C
72-240 hours
0-60 C
72-240 hours
5-150 C

Middlemen are the persons who lend the fishermen money and cover their product
The trading establishments buy fish from the middlemen and supply to the factory

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Fishermen (way 2)

Fishermen (way 3)

Fishermen (way 4)

1 t1

1 t1

t5

Figure 3: Four ways

from catch to facto ry
: Time
t: Temperature

Middlemen

Trading establishment

t4

t3

t2

6, t6
Fishermen (way 1)
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Fish processing plant

Trading establishment
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Catching

Sorting

Washing

Icing

Trading on sea

Unloading

Trading on land

Pre-processing

Receiving

Each handling step affects the quality of

fish (Figure 4, Table 5). Already at the
catching stage the fish can lose quality by
the fishing method. Bottom trawling for a
long time can damage and scratch the fish
and it can be shocked and stressed for too
long before dying. The ambient
temperature in the summer can be 30
35C. In addition, handling and chilling
by ice is carried out slowly and late,
leading to temperature rise. Unloading is
carried out only by human labour, so it is
time consuming. Ice for chilling the raw
material is usually not sufficient, which
results in a temperature higher than 4C.
Some middlemen or trading
establishments carry out re-handling and
pre-processing raw material, where the
raw material is made better in appearance
in spite of poor quality. The containers
used for storage and transportation are
not always suitable as they are not
specialised fish containers. These
containers have poor insulation and the
fisherman has to use more ice for the
preservation of fish. This causes
uncertainty in the shelf life and the
products quality.

Figure 4: Flow chart for fish in Vietnam.

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Table 5: The identified risk as low, medium and high in each handling stage of raw
material in the flow chain.
Problems in each stage

Autolysis

Bacteria

Oxidation

Medium

Low

Low

Mechanical
damage
condition
High

2. Sorting

High

Medium

Medium

High

3. Primary washing

High

Low

Medium

High

4. Icing and putting into

the box

Medium

High

Low

Medium

5. Trading and
transporting at sea

Medium

High

Medium

Medium

6. Unloading

Medium

High

Medium

High

7. Trading and
transporting on land

Medium

Medium

Medium

High

8. Re -icing and preprocessing

Medium

Low

Low

Low

Low

Low

Medium

High

1. Catching method

9. Receiving at factory

3.1.1

Catching method

The main problems are autolysis and mechanical damage. The fish breaks, is bruised
or has scratched skin and the fish is stressed. Some other problems related to physical
conditions are significant like extraneous matter, foreign bodies or hookers. There are
a lot of different fishing methods like trawling, purse seine, gillnetting, lift net and
long line. These methods can influence the gravity of fish. If fishermen carry out
bottom trawling for a long time at certain gravity the fish can press each other causing
breaks in the flesh, bruises and scratches on the skin. In addition when the fish is
broken the outside organism can have easier access to the fish flesh. The catching
process commonly takes about 7-8 hours or more, so the fish is seriously stressed
before death. At that time the glycosis phenomenon happens and lactate acid is
produced rapidly. This causes the rigor mortis process to be shortened leading to
quality defects. The raw material temperature is similar with seawater temperature
(20-25C). This is a good temperature for the development of bacteria.
3.1.2

Sorting

The main problems are autolysis, bacteria growth, oxidation, and mechanical damage.
The fish is crushed, fish temperature is high. Other hazards are smaller, like
extraneous matter (physical) or chemical like lubricant contamination, but this does
not often happen. Most of the vessels in Vietnam are wooden and not very big, which
makes it difficult to use modern automatic systems like conveyer belts or a crane on
board. On board, the fish is spilled out to the board of the vessel and later piled up to

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a bulk or mass for a period of time. Although the fishermen try to sort the fish quickly,
the time for sorting usually takes 2-3 hours. Therefore the raw material mass
temperature stays relatively high or even increases, giving good conditions for growth
of spoilage bacteria and autolysis. Unsuitable sorting tools are often used by the
fisherman like iron rakes, which can cause mechanical damage of the fish. When the
fish handling causes physical damage (mechanical stress), cells can be ruptured and
this enables the autolytic enzymes to react with substrates and produce some spoilage
substances. When the fish is stored in bulk, the temperature increases. This facilitates
enzymatic protein reactions which produce substances like low molecular weight
peptides and free amino-acids. These substances create a good environment for
growth of micro-organisms. In addition, the sorting process is carried out in the open
air in sunshine and wind. Those conditions favour the oxidation process as well as
autolytic and bacterial spoilage. The fish temperature is too high especially bulkstored in the summer and exposed to the wind and sunshine (30-35C).
3.1.3

Primary washing

The problem here is mechanical damage due to strong flushing, autolysis and
oxidation. The fish is put into a plastic basket and then washed by spraying strongly
with water. The fishermen move the fish by throwing the basket with fish inside,
risking bruises or breakage, similar to the sorting stage. The temperature of the
washing is high as this is normally the vessel engine cooling water. This favours rapid
autolytic and bacterial spoilage rates. The fish temperature is still 30-35C and the
time can be long (1-2h) if the catch volume is big.
3.1.4

Icing in boxes

Mechanical damage, autolysis and bacteria are the main quality risks at this stage.
Boxes are often unclean, the fish is put into the box in a wrong way, icing is delayed,
and polyethylene (PE) bags full of fish are piled up. Fish can be crushed by the ice or
by stuffing so this affects the edibility and filleting yield seriously and stimulates
autolysis. In the case of the fish plastic box, the ice is sufficient in the beginning but
soon starts to melt especially at the sides of the box. Parts of the fish are then exposed
to air, resulting in a temperature increase and drier fish. Ice and fish can become one
integrated block which is easily subject to chemical damage in the transport process.
Sometimes the fish is not chilled by ice immediately, so temperature is high for a
prolonged time period and even increases, especially when the fish is piled up to bulk.
Some boxes are used with a lid made of corrosive material which contaminates the
fish. In all these cases above, the fish container is important. The container may be a
PE bag, plastic box, tin box, styropore (foam) box or a bamboo basket. Commonly
the fish box is made of plastic without insulation or made of tin with styropore
insulation foam. The dimensions of the plastic box are 510x130x350 mm and can
hold about 15-20 kg of fish, while the tin and styropore box is usually bigger. The
low value fish caught is normally kept in a plastic tank. As most of the box types are
not well insulated, the raw material can not be maintained at a low temperature. This
leads to rapid growth of bacteria and bad quality. The fish boxes in Vietnam are
usually made of non-sustainable material, providing poor physical protection for the
fish during transport. The boxes are not specially designed for easy cleaning and the

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raw material may be contaminated (Tam et al. 2004). However the temperature of
fish can be maintained in around 0C but the time sometimes is too long (10 days).
3.1.5

Trading and transporting at sea

The problem here is mechanical damage, autolysis and bacteria due to delayed icing.
The catching trip is usually long especially for the big vessels where the time can be
one month or more. Therefore the fish is transferred to another vessel that is going to
land. In this case, care is not always taken to provide enough ice on the fish. The
transfer is usually carried out carelessly leading to mechanical damage. The
temperature in this stage is 0 - 10C and the time is around one day.
3.1.6

Unloading

The problem here is also mechanical damage, autolysis, oxidation and bacteria due to
delayed icing. The unloading process usually takes a long time as this is carried out
by artisanal labour. The ice melts very fast especially in the summer and there is no
supplementary ice. The temperature at this stage is 0 - 10C and the time is around 2
3 hours.
3.1.7

Trading and transporting on land

The problem here is mechanical damage, autolysis, oxidation and bacteria due to
delayed icing and unsuitable transportation facilities. The fish box is still made of
material that is very difficult to clean e.g. bamboo baskets. Crushing of the fish by ice
or by the other fish usually happens in the weighing and transportation process. The
raw material is sometimes exposed to the air, wind and sunshine with high
temperature. The temperature in this stage is 5 - 15C and the time is around 1 2
hours.
3.1.8

Re-icing and pre-processing in the trading establishment or middlemen

The problem here is autolysis and oxidation physical damage. The transport and
handling is carried out too carelessly. In the trading establishment the fish can be preprocessed e.g. headed, gutted, scaled and, washed. Sometimes it is soaked in water or
brine with added oxidants or hydrogen peroxide (H2 O2 ) for bleaching. This makes the
fish look fresh but in reality its quality has seriously decreased. The fillet pieces can
become discoloured to yellow after freezing. The hygienic conditions in the trading
establishment handling area are sometimes not acceptable which can lead to bacterial
contamination. The temperature at this stage is 0 - 6C and the time is around 12 24
hours.

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3.1.9

Reception at the factory

The problem here is mechanical damage due to careless practices. At the reception
area transportation is carried out carelessly. In some factories the fish raw material is
soaked again in water with salt and antioxidants or big block of ice are used, that can
crush the fish. The temperature at this stage is 0 - 4C and the time is around 1 2
hours.
3.2

Present situation of fish handling in Iceland

Catching

Boarding

Holding

Sorting

Bleeding

Gutting

Washing

Chilling

Figure 5 presents a flow chart from a

typical Icelandic vessel with some main
stages of advanced fish handling. The
human labour force is minimal and the
whole process is carried out with the
helping of machines. This procedure is
applied on an Icelandic vessel (trawler)
that captures about 80 tons of fish per
fishing trip. The crew on the vessel is
only 16 people, of which the workers
are 10 persons. Standard operation
procedures for fish raw material are
applied for maintaining fish quality as
long as possible. The handling on board
is not only focused on preventing fish
spoilage but also pays attention to the
hazards related to food safety.
Depending on certain conditions
(catching area, distance from catching
area to land, species of fish, the size of
the vessel, time of the catching trip,
demand from market) some stages or
their order in Figure 5 can be changed a
little bit. Similarly some activities can
be carried out by human labour or
machine, depending on conditions.

Storing

Landing

Figure 5: Flow chart for handling of fish and processing in a typical trawler in Iceland
catching mainly cod and haddock.

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3.2.1

Catching method

In Iceland trawling is the main fishing gear (Figure 6) but the time of trawling is not
very long or around 3-4 hours. If the haul size is big the fish can be pressed, so there
might be some risk of mechanical damage. In addition, the time of the autolysis
process is also reduced. The fish caught by bottom long- line and purse seine gear can
give better quality. In general the time keeping fish in the net should not be very long.
Shorter time reduces the amount of shocked, stressed or dead fish in the net.

Bottom longline
Bottom trawl
Pelagic trawl
Purse seine
Other

Figure 6: Catch of trawler by fishing gear in Iceland 2005 (Statistic Iceland 2005).
3.2.2

Boarding

Tackles are used for transferring the catch from the gear to vessel or hauling in the
trawl. Then the net bottom is opened and the fish can fall down into a steel container
below.
3.2.3

Holding

This stage is carried out especially when the volume of fish caught is quite big. The
fish is put into a tank with chilled seawater (CSW) that includes ice and seawater
slurry or refrigerated seawater (RSW). This stage is very important because it lowers
the fish temperature rapidly and limits the activities of enzymes and bacteria.
Depending on conditions, the sorting stage can be carried out after or right before this
stage. The fish can be boarded by the tackle or using the pump to transfer the fish into
the tank with CSW or RSW. This work is carried out by machine in order to save
time and worker labour. The suitable tools can be used for transferring fish into the
tank with care to avoid mechanical damage. In Iceland this work is carried out very
fast. The fish is poured from the net into the big hold and then the fish is transferred
by conveyer to the sorting stage.

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3.2.4

Sorting

The fish is sorted quickly by hand parallel to the processing stage (see sub- section
3.2.5). This work is carried out below the deck with a conveyor belt. The by-catch is
sold in an auction market on shore.
3.2.5

Bleeding and gutting

In order to the make the fish fillet maintain a good appearance, the fish has to be bled.
Blood stains are regarded as defects, as the fillet should be white. Gutting removes
the fish intestines limiting access of most spoilage bacteria. However, for small fish
like pelagic species, the bleeding and gutting stage is not carried out. But for some
lean fish gutting always is carried out. Then the CSW system is used to decrease fish
temperature quickly.
3.2.6

Washing

After bleeding and gutting the fish is transferred to the washing stage. This stage
cleans blood and viscera residues. The washing stage is carried out in a tub with ice
and seawater. It is carried out quickly in order to avoid losing the nutrition substances.
3.2.7

Chilling

The fish is cooled down by liquid ice, therefore the fish temperature decreases very
fast. This stage is short, about 30 minutes. The liquid ice has a lot of advantages to
flake ice including high regular size, larger surface square and the ability to fill the
entire tub/box and cover the fish. The disadvantage of liquid ice affects the taste of
the fish a little bit because it is made by sea water it can make the fish salted. The
liquid ice is also relatively expensive.
3.2.8

Storing

The fish is iced and arranged in layers in insulated tubs with tube ice for storing. The
tubs are stacked in the hold and are easy to lift by crane when landing. A label is
attached to each tub for traceability at further stages of the process.
3.2.9

Landing to plant or auction market

The raw material is unloaded from the boat by a crane. Transport and weighing is
carried out quickly and carefully. In case of landing to land (e.g. Brim vessel) the raw
material can be processed immediately. In other cases the fish will be transferred to
the auction market. In Iceland there are several auction markets e.g. Fiskmarkadur
Islands. Most of the catch traded there are from fishing trips lasting one day, so the
fish is quite fresh. The catch is sold on a daily basis before landing. At reception the
fish is inspected for sufficient ice and arrangement in the tubs. The fish temperature is
also measured and recorded. Then the fish is size graded and weighed, put in the tubs
with fresh ice and dispatched in the evening of catch date. An internet auction where
buyers can log in and participate from anywhere in the world has been established.

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MATERIALS AND METHODS

Two experiments were carried out including a) determination of the ice melting rate
and b) quality comparison between fish stored in an insulated plastic box and in an
open box without insulation. The open box without insulation was chosen to imitate
boxes commonly used in Vietnam. The first experiment was carried out to determine
the effectiveness of different types of fish boxes in terms of keeping the contents cold.
Based on the first experiment, the best and worst fish boxes were found. The second
experiment was a comparison of fish quality in these best and worst scenarios.
4.1

Experiment for determining the insulation ability

Five types of containers were chosen for the experiment: two types of Splast tub 70
l with lid and without lid and two types of Splast cooler 65 l with seal and without
seal; and the plastic box regarded similar to a typical fish box in Vietnam (VN box)
(Figure 7). The Splast container commonly has some advantageous properties like
durable, good insulation layer with the closed lid, comfortable bottom hole for water
drainage, and easy to clean.

(a)

(b)

(c)

Figure 7: The types of box/tub used for experiment (a) Splast tub 70 l, (b) Splast
cooler 65 l, (c) Vietnamese- like box (VN box).
Determination of ice melting was carried out by measuring the amount of ice after
each period (time recorded). At first all the tubs/boxes were weighed. The grinded
tube ice was filled into all the containers and each one weighed again. At each
measuring point, melted ice was removed by opening the bottom hole and then the
box and tubs weighed. Results were recorded and applied according to the formula:
Mi = Mio - K. t (Huss 1994)
In which:
Mi: kg of ice left in the tub
Mio: kg of ice at the beginning
t: time (hours)
K: melting rate (kg/hour)
The time (t) and Mi, Mio is found by weighing and recording the time. Therefore the
amount of ice melt ing every hour can be calculated.

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4.2

Experiment for the quality change in fish

4.2.1

Fish preparation

Small size cod was used for the experiment. It was caught by the Brim trawler
Hardbakur on 3 January and sampled on 5 January. The fish was gutted and stored in
ice onboard until sampled at the Brim raw material storage. The ice type was tube ice
supplied by Brim. Splast 70 l tub and the VN box were chosen for storing fish in the
experiment because these boxes or tubs resulted in the highest and lowest ice melting
rates based on the previous experiment. The fish and tube ice were put in layers into
the box and tub as follows: VN box 1 layer of fish, 2 layers of ice, Splast tub 2
layers of fish, 3 layers of ice (each layer was around 10 cm); a lid was secured on the
Splast tub. These two fish boxes and tubs were transferred from the Brim Company
to Akureyri University and placed in the wet laboratory room of the University MRI.
Room temperature was chosen for storing the boxes to create conditions more similar
to Vietnam.
4.2.2

Measurements

Two fishes from each container were taken each time of sampling until all the ice had
melted. Three methods were chosen for evaluation: microbiological, chemical and
sensory. Each method is described in details in Appendix 1. The experiment schedule
is shown in Table 6. All the analysis methods (sensory, microbiology and chemical)
are following the IFL procedures described in Appendix 1.
Table 6: Sampling schedule for evaluation.
VN box
Day 0
Day 1
Day 2
Day 3

Splast box
X

X
X
X

Day 4
Day 5
Day 6

Day 7
Day 8
Day 9

Day 10
Day 11
Day 12

Day 13

The sensory evaluation followed the QIM method (form in Appendix 1, Table A)
(Martinsdottir et al. 2001). The panel included three people. There are five main
criteria for evaluation: appearance, eyes, gills, bloods and fillets. Every main criterion

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includes some more detailed criteria. The score for each criterion is from 0 to 3.
Lower scores signify higher quality and the total score can show the general fish
quality.
The micro-biological evaluation is based on the pours plate method (Appendix 1).
Samples from individual fish were bacteriologically assessed for aerobic plate count
using plate count agar and iron agar. This medium was used in order to isolate
spoilage bacteria that produce H2 S. Then the petri dishes were put into the incubator
for incubation at 22C for IA media and 30C for PCA media for three days.
The chemical evaluation was a determination of TVB-N following the Kjeldahl
distillation method (Appendix 1).

RESULTS AND DISCUSSION

The results include experiment results for ice melting and experiment results for a
quality comparison between fish stored in the Splast insulation plastic box and the
Vietnamese-like box. The first experiment was carried out to determine the
effectiveness of different types of fish boxes. Based on the first experiment the best
and worst fish box according to ice preservation was found. The second experiment
focuses on comparing fish quality in the best one (Splast box) and the worst one
(Vietnamese- like box) in order to make validations of the guidelines for handling and
preservation of fish raw material in terms of keeping a low temperature.
5.1

Ice melting experiment

Weigh of ice (kg)

The change of ice left in the box after certain amounts of time is shown in Figure 8.

50
45
40
35
30

VN box
Splast box with lid
Splast box without the
lid

25
20
15
10
5
0

Splast cooler with the

seal
Splast cooler without
seal
0

50

100

150

200

Time (hours)

Figure 8: Weight of ice per container by time for the five different containers.

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The ice in the Vietnamese- like box melted (Figure 8) fastest in comparison with the
other fish boxes. Figure 8 shows that the graph can be estimated by a linear model
and the amount of ice melted per hour is around 0.5 kg because the equation for the
line is y = -0.5482x + 26.435 (Figure 14 Appendix 5). Therefore we can calculate
that after 48 hours (two days) the ice will be fully melted based on this formula.
Weight measurements are most fluctuant (R2 value is lowest) in the VN box.
In the Splast insulated plastic box with lid, the amount of ice melted each hour is
around 0.12 kg according to the linear equation y = -0.1222x + 51.338 (Figure 15
Appendix 5). This is the lowest rate in comparison with the others i.e. this type of box
has the highest insulation ability. Therefore from the formula we can calculate that
after 428 hours (approximately 18 days) the ice will be fully melted. For the Splast
insulation plastic box without the lid the amount of ice melted per hour is around 0.31
kg and the equation is y = -0.3069x + 51.152 (Figure 16 Appendix 5) i.e. after 167
hours (approximately seven days) all the ice will be melted. For the Splast plastic
cooler without the lid the amount of ice melted per hour is around 0.16 kg and the
equation is y = -0.1594x + 49.713 (Figure 17 Appendix 5) i.e. after 312 hours
(approximate 13 days) all the ice will be melted and for the Splast plastic cooler
without the seal on the lid the amount of ice melted each hour is around 0.15 kg
(Figure 18 Appendix 5).
In both coolers the rate of melting is most stable in comparison with the others. The
reason may be that the hole for water drainage is smallest so the influence from
ambient temperature is minimised. In general the Vietnamese-like box is the worst
type and the Splast box with the lid is the best one (Figure 9). Therefore these two
were chosen for the second experiment.

Melting rate of ice (kg/h)

0,6
0,5
0,4
0,3
0,2
0,1
0
Vietnamese-like Splast box Splast cooler Splast cooler Splast with
box
without the lid with the seal
without the
the lid
seal

Figure 9: Melting rate of ice in box used in the experiment.

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5.2
5.2.1

Difference of fish quality in the worst and best scenario case

Sensory quality

The sensory experiment shows that fish stored in ice in the VN box has started to
spoil nearly after three days of preservation (Figure 10). On the third day the average
QIM method score is around 16 and all the ice was melted. Therefore the experiment
had not been carried out. Following Martinsdottir (2001) the fish is not considered fit
for human consumption when the score is higher than 18. The line can be presumed
to have extended to the unacceptable limit after 4.5 days.

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Figure 10: Sensory score (QIM) of fish stored in a VN box and Splast box.
Regarding the fish stored in the Splast insulated tub the quality of fish is maintained
for longer periods of time. After 13 days of preservation the fish was unacceptable
according to the QIM criteria and all the ice was melted. According to Huss (1995)
the shelf life can be predicted at 15 days. This means that the results corresponded
with Husss evaluation because at that time the total days preservation by ice was 15
days (including two days on the boat and 13 days of experiment).
It is worth noticing that the fish quality score at the beginning is about 1 on the QIM
scale, i.e. it is very high quality. The fish is caught just two days before and kept at
very good conditions. Therefore the Splast insulation box can store fish better than
the VN box as predicted.

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5.2.2

Chemical analysis

Figure 11 shows the change in TVB-N during storage in ice.

Figure 11: TVB-N contents of fish in ice stored in VN and Splast containers.

The increase is very slow for the first days. This corresponds with the conclusion of
Oehlenschlager (1992) TVB-N remains constant for the first days of storage or
increases slowly but rises fast later in the spoilage process. The reason may be
explained that the main process leading to spoilage is autolysis (this is corresponding
with the results of microbiology). The fish TVB-N value at the beginning is even
higher than after one day, but the deviation between the two individual fish samples
was also high. So this can be considered as normal deviation in between individuals.
The experiment with the VN box should have been carried out for a longer period for
clearer results. But at that time all the ice had melted.
After three days ice storage the value of TVB-N is still very low in the Splast tub,
so the fish quality is maintained longer like predicted. The TVB-N value does not
increase much until around 10 days of storage, but then it develops quite fast to
around 33 mg TVB-N/100 g fish flesh on day 14. The limit for TVB-N given by Huss
(1994) is 30 mg/100 g fish flesh and according to that the fish has reached the
unacceptable limit. The trend of these curves shows that it is highly likely that the VN
curve will rise sooner than the Splast curve.

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5.2.3

Microbial analysis

Figure 12 shows the change in total bacterial count during storage.

Figure 12: Total bacteria counts (PCA, 30C) in the VN box and the Splast box.
The amount of cfu (colony forming units) increased very fast on the second day of
storage (Figure 12) for the fish stored in the Vietnamese- like box. The qua lity of the
fish sample at the beginning is even higher than the fish on the first day (with PCA
media) and days 1 and 2 (with IA media). This can be explained because the fish is
taken randomly so there are some small differences in quality between individuals.
The amount of colonies incubated in PCA media is less than in IA media. This is
reasonable because the IA media is richer in nutritional substances than PCA media.
In addition the temperature incubation for IA media is 22C, which is lower than for
PCA media (30 C) so conditions are better for psychrophilic spoilage bacteria
growing (Gram 1992) (Figure 12 and 13).
Regarding the significant spoilage organisms (SSOs) with black colour on IA media
at day 13 the total count is about 105 - 106 cfu/g (Figure 14).

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Figure 13: Total bacteria counts (IA, 22C) in the VN box and the Splast box.
In case of the fish stored for a long time like storing in the Splast tub the bacteria
can grow up to around 106 107 cfu/g. The experiment with the VN box should have
been extended for a few days until the bacteria growth became higher than the limit.
Following Huss (1994) the fish can be considered unacceptable when the total
bacterial count is 107 cfu/g.

Figure 14: Black colony counts (IA, 22C) in the VN box and the Splast box.

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Thus for all three evaluation methods the results show that if the fish is stored with
ice in an insulated container like the Splast tub with lid, it can for hold up to 13 days.
It is obvious that the fish quality decreases slowly in the Splast tub but rapidly in the
VN box preservation.

RECOMMENDATIONS FOR IMPROVEMENTS IN VIETNAMESE

FISHERIES

In Vietnam, some fishermen and people involved in the handling of raw material do
not have sufficient knowledge of the importance of raw material quality. Avoiding
spoilage is one part of food safety assurance. Therefore the fish handling activities
must have the objective of maintaining maximum quality. It is important to keep a
continuous work flow and avoid any delay of icing i.e. the temperature and time is
always controlled. The understanding of quality importance should be strengthened
with the people involved in fresh fish ha ndling. They need to be trained and educated
on good handling practice and preservation. Below are some guidelines on the
handling and storing of fish raw material to preserve quality.
Catching the fish can be affected by fishing gear e.g. bottom trawling carried out for a
long time can result in damaged or stressed fish. The fishermen should reduce the
trawling time. A bad odour can appear in fish caught in some specific sea areas.
Avoid catching in these areas. These off-odours can be originated from the food
supply of the fish or some specific bacteria in the fish or from sea pollution e.g. oil
released from oil transportation vessels. In Vietnam today the tackle is only used to
draw the catch on board. This process should be performed carefully to avoid
physical damage to raw material.
Chilling on board often starts late. This leads to temperature increase in the raw
material and exposure to sunlight. If the fish is poured out on to the board floor it can
be affected both by high temperature and physical damage. The vessel floor should
not contact the raw material directly. A layer of clean plastic or rubber can be used
for covering it. This can avoid heat transfer from the boat floor and contamination
from the wooden floor.
Sorting should be carried out quickly. Avoid direct sunlight and wind because it
increases the fish temperature very fast. In addition the fish can dry on the surface due
to the wind, leading to loss of value. Use specialised and clean plastic tools for
sorting. Avoid use of hard corrosive tools like iron hooks to sort the fish. This can
damage the fish. Keep all equipment clean. Consider the use of transport facilities
(conveyers) for saving time at this stage.
If the fish is not gutted at this time, the temperature should be lowered as soon as
possible. Chilled sea water (CSW) is very suitable for this purpose. It is always
available with ice and can lower fish temperature very fast due to large surface heat
transfer. The fish should preferably be put into insulated plastic tanks or tubs e.g. the
Splast tub type with the slurry made by ice and sea water. Flake- ice or grinded

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block ice with sea water should be used. Big block ice should not be used, as the
blocks sharp edges can damage the fish physically. It can also press the fish too
much and decrease the temperature slowly. The NaCl and some other dissolved
mineral substances in the sea water contribute to lower the temperature further. This
activity should not go on for extended time, because the fish can lose nutritional value.
For some fish species quality and shelf life can be reduced much more if they are not
gutted and the viscera removed. E.g. gutting should be carried out for lean fish and
big fish like mackerel and tuna. However for the small size fish species, gutting will
take a lot of time, leading to delayed icing. Washing should be carried out quickly
with low water temperature. The CSW system can be used for washing, which will
lower the fish temperature as well. Avoid flushing water strongly.
There are two methods of icing raw material: direct contact icing, which is often
applied to fish and shrimp and non-contact icing which is often applied to squid,
cuttlefish and octopus. The raw material and the ice are arranged in the fish box layer
by layer. Each layer of fish should not be too thick. The fish layer in the tubs should
not be too thick, as it will slow down the chilling. One ice layer can be put in the
bottom and one on the top and one or two layers of fish in between. The icing is an
important stage and care should be taken to bring enough ice for the whole fishing
trip and make sure the icing in each container is sufficient because it is difficult to add
more ice during the trip. If ice is added, it is only on the top of the box. The heat
transfer can be uneven if the ice melts too much on the sides or bottom of the box, the
fish temperature can go up although plenty of ice is still on the top. Liquid ice can be
an option to consider for Vietnam in the future
The fish should be stored in insulated plastic boxes or tubs. This can help make the
ice last longer and save the place for storing fish. Handle carefully while putting the
fish into the box. Polyethylene bags should not be used to store the fish unorganised
in the chilling hold. Systems for conveying fish are desirable to avoid physical
damage to the delicate fish tissue. Avoid using lids made of corrosive material; it can
contaminate the raw material. The box or tub should be designed for easy stacking or
organising without pressure to the fish to save storing space. For traceability in
further stages of the process it is necessary to label each tub or box.
In the chilling hold plastic trays or small boxes can be used to replace the PE bag.
Systems for conveying or craning fish are optimal to unload the fish from the vessels
to avoid damage and save time. Unloading time will shorten if it is performed by
crane. It is very convenient and should be applied in Vietnam in the future.
The premises of middlemen or trading establishments must assure hygienic
conditions. In the future, auction markets could be an option to cut some middle
stages.
At the factory reception, systems for conveying or trolleying fish are preferable to
avoid physical damage to the fish tissue.

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CONCLUSIONS

There are a lot of stages in the fish raw material flow chain in Vietnam. In each stage
some problems have been identified. There are some defectives that usually occur in
the process from catching on board through a lot of middle stages. At the end in the
factory the raw material quality has decreased. Iceland is a developed country in
fisheries. Based on experiences from Iceland and the present situation in Vietnam
guidelines have been written in order to contribute to the improved fresh fish
handling situation in Vietnam and through that the quality of raw material can be also
improved.
In general the main problems in Vietnamese handling of fish are poor chilling; and
thus a lot of fish are already unacceptable at landing. All the activities should be
carried out more quickly but carefully and tenderly to avoid crushing the fish; keep
the fish always in low temperature conditions (around 0C) and in hygienic
conditions.
It is should be seriously considered to use insulated plastic boxes or tubs. Insulated
tubs are better than open thin boxes for keeping fish; prolonged shelf life is very
valuable. The boxes/tubs like the Splast insulated plastic tubs can be applied in
Vietnam. All the results and the analysis above (see Section 5) show a big difference
between the Vietnamese- like box and the Splast insulation tub. Therefore the worst
and best scenario for storing fish here is storing fish in Vietnamese-like box and
Splast tub with the lid. The experiments for validation of the guidelines in term of
keeping low temperature show that as predicated fish can be stored in the Splast tub
for extended time (up to 13 days). The fish stored in the Vietnamese- like box, on the
other hand, lost quality rapidly.
It is very suitable in practice for storing the fish to chill it in ice. Ice chilling of raw
material has to be improved throughout the process chain. For the catching vessel the
time for the catching trip should be shortened. It should not take a lot of time (10 days
or more). In Iceland the trip only takes 5-6 day at sea.
Training employees is an underestimated problem and should be prioritised. The
captain should be trained to realise assurance of fish quality and he has responsibility
to train again his workers. Avoid soaking raw material for personal profit because the
quality will decrease in reality. This is economic fraud. It is necessary that the
premises should be upgraded to ensure hygienic conditions.
Auction markets might be an option for better control of raw material flow. In the
near future the middle stages sho uld be replaced by a fishing auction market. This can
solve a lot of problems because at that time the fish quality is corresponded with its
value and its price, i.e. the fishermens profit is clear. This leads to higher quality and
more profit. This is a motivation for the fishermen to realise how important keeping
the fish quality is.
More automation is favourable for future improvements in Vietnamese fisheries. The
machine (e.g. conveyer, crane) is considered to gradually replace human labour or

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artisanal activities. For the middle stages (middlemen, trading establishment) the
activities to handle the fish should be mechanised (e.g. using a trolley) for saving time
and human labour. In the future the design of equipment on the vessel should be
changed to replace the wooden or corrosive surface contacting the fish by the
stainless steal material to keep it hygienic and avoid contamination.

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ACKNOWLEDGEMENTS
I would like to thank and acknowledge the valuable assistance of the following
individuals, staff and agencies:
UNU-FTP including Mr. Tumi Tmasson, Mr. r sgeirsson and Mrs. Sigrur
Ingvarsdttir for wholly supporting all the fellows;
My supervisors Prof. Hjrleifur Einarsson and Ms. Arnheiur Eyrsdttir ;
The Marine Research Institute (MRI);
The Icelandic Fisheries Laboratories (IFL);
The University of Akureyri Iceland.

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LIST OF REFERENCES
Botta, J.R. 1995. Evaluation of seafood freshness quality. VCH Publisher. New York.
pp 30-31
Brox J. 1984. Planning and engineering data. Containers for fish handling. FAO
Fish.Circ.
Connell J.J. 1995. Control of Fish Quality. Fourth edition. Fishing News Book.
Austria. pp 37-64
Frederiksen M., Popescu V., Olsen K.B. 1997. Integrated quality assurance of chilled
food fish at sea. Seafood from producer to consumer. Integrated approach to quality.
Developments in Food Science 38, pp 87 - 96
Gram L. 1992. Evaluation of the bacteriological quality of seafood. Quality
assurancein Food industry. pp 269-282
Gram L, Huss H.H. 1996. Microbiological spoilage of fish and fish products.
International Journal of Food Microbiology 33. pp 121-137
Hobbs G. 1982. Changes in fish after catching. Fish handling and processing. Torry
Research Station. pp 20-27
Howgate P.F. 1982. Fish handling and processing. Torry Research Station pp 20-42
Huss H.H. 1994. Assurance of seafood quality. FAO fisheries technical paper334 . pp
46-53
Huss H.H. 1995. Quality and quality changes in fresh fish. FAO fisheries technical
paper 348. pp 35-67
Jonsdottir S. 1992. Quality Index Method and TQM-System. Quality issues in the
Fish industry. The research Liaison Office. University of Iceland. pp 81-94
Jrgensen. 1965. Hygienic aspects of fish boxes. Fish handling and preservation.
Organisation for Economic cooperation and development. pp 213-231
Kelman J.H. 1982. Handling wet fish at sea. Fish handling and processing. Torry
Research Station. pp 28, 29
Kolakowska A. 1992. Effect of fishing season on shelf- life of iced Baltic herring.
Quality assurance in Food industry. pp 81-92
Liston J. 1992. Bacterial spoilage of seafood. Quality assurance in Food industry. pp
93-105

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Love R.M. 1982. Basic Fact about fish. Fish handling and processing. Torry
Research Station. pp 2-19
Martinsdttir E., Sveinsdottir K., Luten J., Schelvis-Smit J.B., Hyldig R.G. 2001.
Reference manual for the fish sector: sensory evaluation of fish freshness. QIM
Eurofish. P.O. Box. 68, 1970 AB IJumiden, The Netherlands.
Oehlenschlager J. 1992. Evaluation of some well established and some underrated
indices for the determination of freshness and/or spoilage of ice stored wet fish.
Quality assurance in Food industry. pp 339-358
Ryrvik J. 1982. Handling and Transportation of fish. Developments in handling and
processing fish
Tam H.L., Bao H.N.D., Marie E. 2004. Guideline for Handling and preservation of
black-tiger shrimp and cephalopod, Agriculture Publisher
Valdimarsson G. 1992. Handling of fresh fish on board fishing Vessel. Quality issues
in the Fish industry. The research Liaison Office. University of Iceland. pp 68-79
Vyncke W. 1965. Temperature, packing and quality in fish market. Fish handling and
preservation Organisation for Economic cooperation and development. pp 233-255
Wignall J. 1982. Handling wet fish on shore. Fish handling and processing. Torry
Research Station pp 42-55
Food and Agriculture Organization of the United Nation. Fish statistic 2003
<http://www.fao.org>
Ministry of Fisheries of Vietnam. Fisheries Scientific Technological Economic
Information. <http://www.fistenet.gov.vn>
Statistic Iceland. Fisheries. Catch by type of fishing gear and species 2003-2004
<http://www.hagstofa.is>
The Icelandic Fisheries Laboratories (IFL). 2005. Chemical, Microbiological and
Sensory Method to evaluate fish raw material quality

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APPENDIX 1: ANALYSIS METHOD (SENSORY, MICROBIOLOGY AND

CHEMICAL)
1. Sensory method
1.1
1.2

Material
Samples:
Nylon sheet:
Table for evaluation:
Sticky paper for coding:
Evaluation form:

whole fish and fish fillets

1 unit
1 unit
1 unit
3 forms

Method

2 fishes were taken from the box and put into the PE bag and the sample coded. (Then
the 2 samples are taken to the sensory room, put on the table and coded). Three
people from the fellow group carried out the sensory evaluation on whole fish and on
the fillet.
Appearance of skin, firmness of flesh, slime formation, colour and form of eyes and
finally colour, smell and mucus formation of gills was evaluated according to the
Quality Index Method (QIM).

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Table Ia: Sensory evaluation form

Quality parameter

Appearance

Skin

Stiffness

Eyes

Cornea

Form

Colour of
pupil

Gills

Colour

Smell

Muscus

Blood

Fillets

Colour

Colour

Points

Score of each code

Description

Bright, iridescent pigmentation

Rather dull, becoming discoloured

Dull

In rigor

Firm, elastic

Soft

Very soft

Clear

Opalescent

Milky

Convex

Flat, slightly, sunken

Sunken, concave

Black

Opaque

Grey

Bright

Less colour, becoming discoloured

Discoloued, brown spots

Brown, discoloured

Fresh, seaweedy, metallic

Neutral, grassy, musty

Yeast, bread, beer, sour milk

Acetic acid, sulphuric, very sour

Clear

Milky

Milky, dark, opaque

Red

Dark red

Brown

Translucent, bluish

Waxy, milky

Opaque, yellow, brown spots

Quality Index (0-23)

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Microbiological method

2.1

Material
Iron Agar:
Petri dishes:
Glass tube:
Pipette:
Blender:
Homogenizer (stomacher):
Analysis scale:
Alcohol 70%:

2.2

1000ml
40 units
10 units
10 units
1 unit
1 unit
1 unit
1000ml

Method:

The fish is aseptically filleted and minced to 100g. (25g were kept for microbiological
analysis and the remains put into the refrigerator for chemical evaluation) A 1/10
dilution is made by mincing and mixing 25g of mince with 225g of dilution buffer
and homogenizing in the stomacher for 60 seconds to make 1/10 dilution. 2 petri
plates are used for each dilution, 2x1ml are then inoculated into the first pair of plates
(1/10) and 2x0,1ml on the next pair (1/100), and similar with (1/1000) and (1/10000).
Melted 45C Iron agar is poured on the plates, stirred and left to cool. Agar plates are
incubated at 22C fo r 48 hours and the number of colony forming units (c.f.u.) is
counted. Black and white colonies were observed and counted respectively. The
counts were calculated against the dilution factors that contains at least 25 up to 250
colonies (IFL method)

Chemical method

3.1

Material

TVB-N method by steam distillation:

- Aqueous tricloacetic acid 7,5%: preparing 1000ml (each time using:
200ml)
- NaOH 10%
preparing 100ml (each time using: 10ml)
- Acid boric 4%
preparing 100ml (each time using: 10ml)
- Methyl red
preparing 10ml (each time using: 0,04ml)
- Sulfuric acid 0,025N preparing 100ml
- Water distilled
1 unit
- Analysis scale
1 unit
- Waring blender:
1 unit
- Distillator: 1 Kjeldahl-type
1 unit
- Burette
0,05ml graduated
- Flask
250 ml.
- Beaker
250 ml
- Pipette
10 ml.
- Glass cup
100ml

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3.2

Filter paper
Method:

Spoilage chemical indicators are evaluated by measuring total volatile basic nitrogen
(TVB-N) levels in the minced fish species. The determination of TVB-N by steam
distillation is used. In this method:
- 200 ml of 7,5% aqueous trichloroacetic acid solution is added to 100
grams of fish muscle and homogenized in the blender.
- The mixture is filtered by filter paper.
- Steam distillation is carried out using a Kjeldahl-type distillator.
- 25ml of the filtrate is put into the distillation flask and 6 ml of 10% NaOH
is added to it.
- An beaker containing 10 ml of 4% aqueous boric acid solution and 0.04
ml of methyl red and bromocresol green indicator are used for titration of
ammonia and placed at the end of the condenser.
- Distillation is done until the final volume of 50 ml was obtained in the
beaker (40 ml of distillate). This process take about 10 minutes
- The boric acid solution turned green when alkalinized by the distilled
TVB-N. This solution is titrated using a 0,05 ml graduated burette containing
0.025N H2SO4 and complete neutralization is obtained when the colour
turned pink on the addition of a further drop of H2SO4 (IFL method).
Calculation: (mgN/100g):
14mg/mol*a*b*300
25
a: ml of sulphuric acid
b: normality of sulphuric acid

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APPENDIX 2 - SENSORY TESTING RESULTS

Table IIa: The QIM method score for Vietnamese-like box

VN box

Panellists
Ofred

Quang

Average
Yen

Standard
deviation

Day 0
5/1/2006

Fish 1
Fish 2

2
0

0
2

1
1,5

1
1,17

1
1,04

Day 1
6/1/2006

Fish 1A
Fish 1B

2
3

8
2

2
1,5

4
2,17

3,46
0,76

Day 2
7/1/2006

Fish 2A
Fish 2B

11
8

11
14

7
8

9,67

2,31

10,00

3,46

17,00
14,33

5,29
4,04

Day 3
8/1/2006

Fish 3A
Fish 3B

19
18

21
15

11
10

Table IIb: The average value of QIM method score for Vietnamese-like box
VN box

Score average

General average

Day 0
5/1/2006

Fish 1
Fish 2

1
1,17

1,09

Day 1
6/1/2006

Fish 1A
Fish 1B

4
2,17

3,09

Day 2
7/1/2006

Fish 2A
Fish 2B

9,67
10,00

9,84

Day 3
8/1/2006

Fish 3A
Fish 3B

17,00
14,33

15,67

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Table IIc: The QIM method score for Splast box

VN box

Panellists
Ofred

Quang

Average
Yen

Standard
deviation

Day 0
5/1/2006

Fish 1
Fish 2

2
0

0
2

1
1,5

1
1,17

1
1,04

Day 4
9/1/2006

Fish 4A
Fish 4B

8
4

7
8

5,5
4,5

6,83
5,50

1,26
2,18

Day 7
12/1/2006

Fish 7A
Fish 7B

14
9

17
12

18
11

16,33
10,67

2,08
1,53

Day 10
15/1/2006

Fish 10A
Fish 10B

9
11

16
20

14,5
18

13,17
16,33

3,69
4,73

Day 13
18/1/2006

Fish 13A
Fish 13B

20
22

23
21

19,5
20,5

20,83
21,17

1,89
0,76

Table IId: The average value of QIM method score for Splast box
VN box

Average

General average

Day 0
5/1/2006

Fish 1
Fish 2

1
1,17

1,09

Day 4
9/1/2006

Fish 4A
Fish 4B

6,83
5,50

6,17

Day 7
12/1/2006

Fish 7A
Fish 7B

16,33
10,67

13,5

Day 10
15/1/2006

Fish 10A
Fish 10B

13,17
16,33

14,75

Day 13
18/1/2006

Fish 13A
Fish 13B

20,83
21,17

21

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APPENDIX 3 - CHEMICAL TESTING RES ULTS

Table IIIa: The TVB-N value for Vietnamese-like box

VN box

H2SO4 (ml)
First time

H2SO4 (ml)
Second time

H2SO4 (ml)
Average

TVB (mg/100g)

Day 0
5/1/2006

Fish 1
Fish 2

2,7
2,9

2,3
2,8

2,5
2,85

10,5
11,97

Day 1
6/1/2006

Fish 1A
Fish 1B

2,4
2,5

2,5
2,6

2,45
2,55

10,29
10,71

Day 2
7/1/2006

Fish 2A
Fish 2B

2,6
3,1

2,8
2,9

2,7
3

11,34
12,6

Day 3
8/1/2006

Fish 3A
Fish 3B

3,2
3,3

3,2
3,4

3,2
3,35

13,44
14,07

Table IIIb: The Average of TVB-N value for Vietnamese-like box

VN box

TVB (mg/100g)

Average

STDEV

Day 0
5/1/2006

Fish 1
Fish 2

10,5
11,97

11,24

1,04

Day 1
6/1/2006

Fish 1A
Fish 1B

10,29
10,71

10,5

0,27

Day 2
7/1/2006

Fish 2A
Fish 2B

11,34
12,6

11,97

0,89

Day 3
8/1/2006

Fish 3A
Fish 3B

13,44
14,07

13,76

0,45

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Table IIIc: The TVB-N value for Splast box

VN box

H2SO4 (ml)
First time

H2SO4 (ml)
Second time

H2SO4 (ml)
Average

TVB (mg/100g)

Day 0
5/1/2006

Fish 1
Fish 2

2,7
2,9

2,3
2,8

2,5
2,85

10,5
11,97

Day 4
9/1/2006

Fish 4A
Fish 4B

2,6
2,3

2,4
2,4

2,5
2,35

10,5
9,87

Day 7
12/1/2006

Fish 7A
Fish 7B

3,5
3

4
2,8

3,75
2,9

15,75
12,18

Day 10
15/1/2006

Fish 10A
Fish 10B

3,5
4,4

4
4,6

3,75
4,5

15,75
18,9

Day 13
18/1/2006

Fish 13A
Fish 13B

8,2
7

8,5
7,1

8,35
7,05

35,07
29,61

Table IIId: The Average of TVB-N value for Splast box

VN box

TVB (mg/100g)

Average

STDEV

Day 0
5/1/2006

Fish 1
Fish 2

10,5
11,97

11,24

1,04

Day 4
9/1/2006

Fish 4A
Fish 4B

10,5
9,87

10,19

0,45

Day 7
12/1/2006

Fish 7A
Fish 7B

15,75
12,18

13,97

2,52

Day 10
15/1/2006

Fish 10A
Fish 10B

15,75
18,9

17,35

2,23

Day 13
18/1/2006

Fish 13A
Fish 13B

35,07
29,61

32,34

3,86

UNU-Fisheries Training Programme

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APPENDIX 4 - MICROBIOLOGY TESTING RESULTS

Table IVa: Cfu count after 48 hours incubation - VN box and Splast tub day 0
Day 0 (48h)
5.1.2006
VN box
Fish 1
PCA
30C

Fish 2

Fish 1
IA (White)
22C

Fish 2

Fish 1
IA (Black)
22C

Fish 2

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
3
8
11
13

D1/100
0
0
0
0

D1/1000 D1/10000
0
0
0
0
0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

>250
>250
52
40

19
14
2
2

0
1
1
0

0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

Table IVb: Cfu count after 72 hours incubation - VN box and Splast tub day 0
Day 0 (72h)
5.1.2006
VN box
Fish 1
PCA
30C

Fish 2

Fish 1
IA (White)
22C

Fish 2

Fish 1
IA (Black)
22C

Fish 2

D1/100
0
0
0
0

D1/1000
0
0
0
0

D1/1000
0
0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
7
16
11
19

Petri
Petri
Petri
Petri

1
2
1
2

> 250
> 250
73
68

30
29
7
3

0
1
1
0

0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

UNU-Fisheries Training Programme

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Table IVc: Cfu count after 48 hours incubation - VN box day 1

Day 1 (48h)
6.1.2006
VN box
Fish 1A
PCA
30C

Fish 1B

Fish 1A
IA (White)
22C

Fish 1B

Fish 1A
IA (Black)
22C

Fish 1B

D1/100
0
0
0
0

D1/1000
0
0
0
0

D1/1000
0
0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
4
3
0
0

Petri
Petri
Petri
Petri

1
2
1
2

19
22
7
6

0
0
0
0

0
0
0
0

0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

Table IVd: Cfu count after 72 hours incubation - VN box day 1

Day 1 (72h)
6.1.2006
VN box
Fish 1A
PCA
30C

Fish 1B

Fish 1A
IA (White)
22C

Fish 1B

Fish 1A
IA (Black)
22C

Fish 1B

D1/100
0
0
0
0

D1/1000
0
0
0
0

D1/1000
0
0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
10
5
1
2

Petri
Petri
Petri
Petri

1
2
1
2

30
42
10
7

1
2
0
0

0
0
0
0

0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

0
0
0
0

0
0
0
0

0
0
0
0

0
0
0
0

UNU-Fisheries Training Programme

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Nguyen

Table IVe: Cfu count after 48 hours incubation - VN box day 2

Day 2 (48h)
7.1.2006
VN box
Fish 2A
PCA
30C

Fish 2B

Fish 2A
IA (White)
22C

Fish 2B

Fish 2A
IA (Black)
22C

Fish 2B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
8
8
484
> 250

D1/100
0
0
5
169

D1/1000
0
0
13
24

D1/10000
0
0
1
9

Petri
Petri
Petri
Petri

1
2
1
2

40
60
127
129

5
0
7
9

1
0
0
0

0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

0
5
0
2

0
1
0
0

0
0
0
0

0
0
0
0

Table IVf: Cfu count after 72 hours incubation - VN box day 2

Day 2 (72h)
7.1.2006
VN box
Fish 2A
PCA
30C

Fish 2B

Fish 2A
IA (White)
22C

Fish 2B

Fish 2A
IA (Black)
22C

Fish 2B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
8
8
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

40
60
127
129

5
0
7
9

1
0
0
0

0
0
0
0

Petri 1
Petri 2
Petri 1
Petri 2

0
5
0
2

0
1
0
0

0
0
0
0

0
0
0
0

UNU-Fisheries Training Programme

D1/100
0
0
5
169

D1/1000
0
0
13
24

D1/10000
0
0
1
9

49

Nguyen

Table IVg: Cfu count after 48 hours incubation - VN box day 3

Day 3 (48h)
8.1.2006
VN box
Fish 3A
PCA
30C

Fish 3B

Fish 3A
IA (White)
22C

Fish 3B

Fish 3A
IA (Black)
22C

Fish 3B

D1/100
125
112
118
102

D1/1000
16
8
12
8

D1/1000
0
1
0
1
0

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

> 250
> 250
> 250
40

25
44
16
35

4
4
1
2

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

> 250
> 250
38
36

27
32
1
2

3
3
2
0

D1/100
246
142
137
159

D1/1000
25
15
9
13

D1/1000
0
1
1
1
1

Table IVh: Cfu count after 72 hours incubation - VN box day 3

Day 3 (72h)
8.1.2006
VN box
Fish 3A
PCA
30C

Fish 3B

Fish 3A
IA (White)
22C

Fish 3B

Fish 3A
IA (Black)
22C

Fish 3B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

> 250
> 250
> 250
46

49
37
32
19

7
4
6
4

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

> 250
> 250
840
36

31
36
1
3

3
3
2
0

UNU-Fisheries Training Programme

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Table IVi: Cfu count after 48 hours incubation - Splast tub day 4
Day 4 (48h)
9.1.2006
Splast box
Fish 4A
PCA
30C

Fish 4B

Fish 4A
IA (White)
22C

Fish 4B

Fish 3A
IA (Black)
22C

Fish 3B

D1/100
0
0
0
0

D1/1000
0
0
0
0

D1/1000
0
0
0
0
0

250
250
250
250

28
28
22
22

0
0
1
0

0
0
0
0

0
0
3
0

0
0
0
0

0
0
0
0

0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
17
25
17
19

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

Petri
Petri
Petri
Petri

1
2
1
2

Table IVj: Cfu count after 72 hours incubation - Splast tub day 4
Day 4 (72h)
9.1.2006
Splast box
Fish 4A
PCA
30C

Fish 4B

Fish 4A
IA (White)
22C

Fish 4B

Fish 3A
IA (Black)
22C

Fish 3B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
30
39
36
46

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

Petri
Petri
Petri
Petri

1
2
1
2

UNU-Fisheries Training Programme

D1/100
1
3
1
4

D1/1000
0
0
0
0

D1/10000
0
0
0
0

250
250
250
250

40
38
31
32

2
5
2
0

0
0
0
0

3
6
6
2

0
0
0
1

0
0
0
0

0
0
0
0

51

Nguyen

Table IVk: Cfu count after 48 hours incubation - Splast tub day 7
Day 7 (48h)
12.1.2006
Splast box
Fish 7A
PCA
30C

Fish 7B

Fish 7A
IA (White)
22C

Fish 7B

Fish 7A
IA (Black)
22C

Fish 7B

D1/100
36
45
56
43

D1/1000
1
0
1
0

D1/1000
0
0
0
0
0

250
250
250
250

243
231
123
156

21
32
5
9

0
2
0
0

54
63
15
19

1
1
0
0

0
0
0
0

0
0
0
0

D1/100
48
53
67
64

D1/1000
2
2
4
4

D1/1000
0
0
0
1
0

250
250
250
250

> 250
> 250
178
162

27
38
12
13

2
4
1
0

64
71
27
29

4
3
1
1

0
0
0
0

0
0
0
0

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

Petri
Petri
Petri
Petri

1
2
1
2

Table IVl: Cfu count after 72 hours incubation - Splast tub day 7
Day 7 (72h)
12.1.2006
Splast box
Fish 7A
PCA
30C

Fish 7B

Fish 7A
IA (White)
22C

Fish 7B

Fish 7A
IA (Black)
22C

Fish 7B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

Petri
Petri
Petri
Petri

1
2
1
2

UNU-Fisheries Training Programme

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Table IVm: Cfu count after 48 hours incubation - Splast tub day 10
Day 10 (48h)
15.1.2006
Splast box
Fish 10A
PCA
30C

Fish 10B

Fish 10A
IA (White)
22C

Fish 10B

Fish 10A
IA (Black)
22C

Fish 10B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

D1/100
> 250
> 250
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

>
>
>
>

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

5
15
> 250
> 250

D1/1000
> 250
> 250
> 250
> 250

250
250
250
250

>
>
>
>

D1/1000
0
20
19
98
75

250
250
250
250

> 250
32
> 250
> 250

3
1
7
15

0
0
0
1

Table IVn: Cfu count after 72 hours incubation - Splast tub day 10
Day 10 (72h)
15.1.2006
Splast box
Fish 10A
PCA
30C

Fish 10B

Fish 10A
IA (White)
22C

Fish 10B

Fish 10A
IA (Black)
22C

Fish 10B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

>
>
>
>

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

5
16
> 250
> 250

UNU-Fisheries Training Programme

D1/100
> 250
> 250
> 250
> 250
250
250
250
250

D1/1000
664
728
> 250
> 250
>
>
>
>

D1/10000
23
22
116
82

250
250
250
250

624
572
164
153

3
1
8
20

0
0
0
1

53

Nguyen

Table IVo: Cfu count after 48 hours incubation - Splast tub day 13
Day 13 (48h)
18.1.2006
Splast box
Fish 13A
PCA
30C

Fish 13B

Fish 13A
IA (White)
22C

Fish 13B

Fish 13A
IA (Black)
22C

Fish 13B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

D1/100
> 250
> 250
> 250
> 250

D1/1000
> 250
> 250
> 250
> 250

D1/10000
66
64
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

> 250
> 250
125
142

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

11
10
18
15

Table IVp: Cfu count after 72 hours incubation - Splast tub day 13
Day 13 (72h)
18.1.2006
Splast box
Fish 13A
PCA
30C

Fish 13B

Fish 13A
IA (White)
22C

Fish 13B

Fish 13A
IA (Black)
22C

Fish 13B

Petri
Petri
Petri
Petri

1
2
1
2

D1/10
> 250
> 250
> 250
> 250

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

956
39
876
744

Petri
Petri
Petri
Petri

1
2
1
2

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

>
>
>
>

250
250
250
250

14
10
22
20

UNU-Fisheries Training Programme

D1/100
> 250
> 250
> 250
> 250

D1/1000
> 250
> 250
> 250
> 250

D1/10000
82
76
540
432

54

Nguyen

APPENDIX 5 - RESULTS OF ICE MELTING AMOUNT BY TIME FOR

SOME TYPES OF BOX/TUB

Weight of ice (kg)

30
25

y = -0,5482x + 26,435
2
R = 0,9881

20
15
10
5
0
0

10

15

20

25

30

35

40

Time (hours)

Figure Va: Weight of ice in container by time for Vietnamese-like box

60
Weight of ice (kg)

50
40
30
20

y = -0,1222x + 51,338
2
R = 0,9995

10
0
0

50

100

150

200

Time (hours)

Figure Vb: Weight of ice in container by time for Splast box with the lid

UNU-Fisheries Training Programme

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Nguyen

60

Weight of ice (kg)

50

y = -0,3069x + 51,152
2
R = 0,9961

40
30
20
10
0
0

20

40

60

80

100

120

140

160

Time (hours)

Figure Vc: Weight of ice in container by time for Splast box without the lid

Weight of ice (kg)

60
50

y = -0,1594x + 49,713
2
R = 0,9985

40
30
20
10
0
0

50

100

150

200

Time (hour)

Figure Vd: Weight of ice in container by time for Splast cooler with the seal on
the lid

UNU-Fisheries Training Programme

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Nguyen

Weight of ice (kg)

60
50

y = -0,1528x + 48,809
2
R = 0,9992

40
30
20
10
0
0

50

100

150

200

Time (hours)

Figure Ve: Weight of ice in container by time for Splast cooler without the seal
on the lid

UNU-Fisheries Training Programme

57