THE EFFECTS OF MALAY HERBS ON THE STORAGE STABILITY OF CHILLED KEROPOK LEKOR

AZNI A.AZIZ

BACHELOR OF SCIENCE (HONS.) FOOD SCIENCE AND TECHNOLOGY FACULTY OF APPLIED SCIENCES UNIVERSITI TEKNOLOGI MARA

NOVEMBER 2008

THE EFFECTS OF MALAY HERBS ON THE STORAGE STABILITY OF CHILLED KEROPOK LEKOR

AZNI A.AZIZ

Final Year Project Report Submitted in Partial Fulfillment of the Requirement for the Degree of Bachelor of Science (Hons.) Food Science and Technology in the Faculty of Applied Sciences Universiti Teknologi MARA

NOVEMBER 2008

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The Final year Project Report entitled The Effects of Malay Herbs on the Storage Stability of Chilled Keropok Lekor was submitted by Azni A.Aziz, in partial fulfillment of the requirements for the Degree of Bachelor of Science (Hons.) Science and Food Technology, in the Faculty of Applied Science, and was approved by

Assoc. Prof. Dr. Noriham Abdullah Supervisor B. Sc. (Hons.) Food Science and Technology Faculty of Applied Science Universiti Teknologi MARA 40450 Shah Alam Selangor

Pn. Azizah Othman Project Coordinator B. Sc. (Hons.) Food Science and Technology Faculty of Applied Science Universiti Teknologi MARA 40450 Shah Alam Selangor

Assoc. Prof. Dr. Norizzah Abd. Rashid Head of Programme B. Sc. (Hons.) Food Science and Technology Faculty of Applied Science Universiti Teknologi MARA 40450 Shah Alam Selangor

Date:

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ACKNOWLEDGEMENTS

In the name of Allah, The Most Gracious and The Most Merciful. As the Author of this project, I would like to take this opportunity to convey my greatest gratitude and appreciation to all the people who have assisted me during the preparation of this project. First of all, I would like to express my deepest thanks and appreciation to my supervisor, Assoc. Prof. Dr. Noriham Abdullah for her understanding, patience, invaluable guidance, suggestion, and constant encouragement throughout the planning of the execution of this project. Special thanks and appreciation also goes to the Program Head of BSc. (Hons) Food Science and Technology, Assoc. Prof. Dr. Norizzah Abd. Rashid and Project Coordinator Pn. Azizah Othman as well as all the lecturers for their guidance and invaluable advices throughout all the years of my study in Universiti Teknologi Mara, Shah Alam. The insightful information and help is duly acknowledged. Sincere thanks are also extended to the Food Technology laboratory staffs for their informative guidance and support especially to Pn. Siti Mahani, Pn. Nora and Cik Shuhada. I am grateful to Izreen, Nor Azlin Tokiman and her colleagues for their help in part of the experiment and preliminary discussion. Without all of you, this project might not be a success. Not forgetting, my warmest and heartiest appreciation to all my classmates and friends especially Mazuin Che Mahmood, Nurul Akma Muni, Faridah Ahamad, Norjuwariah Mukhtar, Farahana Basri, Wan Norshela Meor Zainol for their kindness and helpfulness in completing the course. Last but not least, my deepest thank to my beloved family, Abah, Mama, Angah, Kak lang, Kak Chik and adik for for their understanding, love, encouragement, financial support and motivation throughout my study.

Azni A.Aziz

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TABLE OF CONTENTS

AKNOWLEDGEMENTS TABLE OF CONTENTS LIST OF TABLES LIST OF FIGURES LIST OF ABBREVIATIONS ABSTRACT ABSTRAK

Page iii iv vi vii viii ix x

CHAPTER 1 INTRODUCTION 1.1 Background and Problem statement 1.2 Significance of study 1.3 Objectives of study

1 1 2 3

CHAPTER 2 LITERATURE REVIEW 2.1 Fish 2.1.1 Fish Muscle 2.1.2 Shortfin Scad Fish 2.1.2.1 Keropok Lekor 2.1.3 Chilled Fish 2.2 Spoilage in Fish 2.2.1 Microbiological Spoilage 2.2.2 Chemical Spoilage (oxidation) 2.2.3 Autolytic Spoilage 2.3 Antioxidant 2.3.1 Antioxidant as antimicrobial 2.3.2 Synthetic Antioxidant 2.3.2.1 BHA and BHT 2.3.3 Natural Antioxidant 2.3.3.1 Naturally occurring Phenolic compounds 2.4 Herbs 2.4.1 Malay herbs 2.4.1.1 Polygonum hydropiper (kesum) 2.4.1.2 Centella asiatica (pegaga)

4 4 6 7 8 9 11 13 15 18 19 22 24 26 27 28 30 32 34 36

CHAPTER 3 METHODOLOGY 3.1 Materials 3.1.1 Raw Material 3.1.2 Chemicals 3.1.3 Instrument/Apparatus 3.2 Sample preparation 3.2.1 Herbs preparation iv

39 39 39 39 39 40 40

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3.3

3.4

3.5 3.6

3.2.2 Keropok Lekor preparation Chemical Analysis 3.3.1 Peroxide Value (PV) 3.3.1.1 Lipid Extraction 3.3.1.2 Peroxide Value 3.3.1 Thiobarbituric acid (TBA) Microbiological Analysis 3.4.1 Total Plate Count 3.4.2 Mould and Yeast Count Sensory Analysis Statistical Analysis

40 41 41 41 42 43 43 43 44 44 44

CHAPTER 4 RESULTS AND DISCUSSSION 4.1 Chemical Analysis 4.1.1 Peroxide Value of chilled Keropok Lekor 4.1.2 Thiobarbituric acid Value of chilled Keropok Lekor 4.2 Microbiological Analysis 4.2.1 Total plate Count of chilled Keropok Lekor 4.2.2 Mould and yeast Count of chilled Keropok Lekor 4.3 Sensory Analysis

45 45 45 48 50 50 53 55

CHAPTER 5 CONCLUSION AND RECOMMENDATIONS

57

CITED REFERENCES APPENDICES CURICULUM VITAE

59 64 79

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LIST OF TABLES

Tables

Page

2.1 2.2

The nutrient composition of oily and white fish Predicted shelf life of fish products stored at different temperatures Nutrient composition of Shortfin scad fish. The causes of the various types of spoilage in fish The recommended limit for bacteria present in a product Solubility of the BHA and BHT in various solvents Amount of total phenolic of various Malay herbs Formulation of Keropok Lekor Mean sensory attributes scores of chilled Keropok Lekor

5 5

2.3 2.4 2.5 2.6 2.7 3.1 4.1

7 13 15 23 34 41 57

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LIST OF FIGURES

Figure

Page

2.1 2.2

Mechanism of Autoxidation Oxidation process of polyunsaturated fatty acids found in fish tissue Mechanism of antioxidant Chemical structure of BHA & BHT Structure of Phenolic compounds Peroxide value of chilled Keropok Lekor Thiobarbituric acid value of chilled Keropok Lekor Total plate count of chilled Keropok Lekor Mould and Yeast count of chilled Keropok Lekor

16 18

2.3 2.4 2.5 4.1 4.2 4.3 4.4

21 27 29 46 49 51 55

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LIST OF ABBREVIATIONS

BB BHA BHT CFU/g CFU/ml DH FTC PG PV SSO TBA TBHQ TMA TMAO TNTC WH

: : : : : : : : : : : : : : : :

BHA/BHT Butylated hydroxyl anisole Butylated hydroxytoluene Colony forming unit per gram Colony forming unit per milliliter Dry Herbs Ferric thiocyanate Propyl gallate Peroxide value Specific spoilage organisms Thiobarbituric acid Tertiary butyl hydroxyquinone Trimethylamine Trimethyl amino oxide Too numerous to count Wet Herbs

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ABSTRACT

THE EFFECTS OF MALAY HERBS ON THE STORAGE STABILITY OF CHILLED KEROPOK LEKOR

Keropok Lekor is a famous traditional cuisine especially in the East Coast of Malaysia. The primary ingredients are fish, flour, sugar and salt. This study was conducted on Keropok Lekor to observe the storage stability for 12 days at chilled temperature (4C) after adding mixed local herbs. The formulation without any incorporation was used as control. The other formulations were incorporation with dry mix Malay herbs, incorporation with wet mix Malay herbs and another one is incorporation with BHA/BHT as a comparison between synthetic and natural plant antioxidant. Analysis that was carried out is chemical analysis which evaluated through measurement of primary (Peroxide Value) and secondary (Thiobarbituric Acid Value) oxidation products. Microbiological determinations included the Total Plate Count and Mould and Yeast Count. Sensory evaluation tests were also carried out. Dry and wet mix herbs of P. hydropiper and C. asiatica resulted in significant (P< 0.05) inhibition of lipid peroxidation as compared to control sample. At initial, the Thiobarbituric Acid Value obtained was not significantly different than to Keropok Lekor that was added with BHA/BHT. Total plate count showed a high count for all the samples. But, the treated samples were significantly lower (P<0.05) than control until day 9 of storage. Mould and Yeast Count however showed a lower count than Total Plate Count whereby control was significantly higher (P<0.05) than the other treated samples throughout the storage time. The highest overall acceptability in sensory evaluation was the Keropok Lekor incorporated with wet herbs. In conclusion, the best antioxidative and antimicrobial effects were obtained from the Keropok Lekor incorporated with BHA/BHT. But the incorporation of Keropok Lekor with natural herbs also effectively reduced the lipid peroxidation which shows antioxidative effect up to 9 days of storage. Study showed that not much antimicrobial effect was observed for the treated samples.

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ABSTRAK

KESAN HERBA MELAYU KE ATAS KESTABILAN PENYIMPANAN KEROPOK LEKOR DALAM KEADAAN DINGIN

Keropok lekor adalah makanan tradisi terkenal di Malaysia terutamanya di pantai timur. Ramuan utama terdiri daripada ikan selayang, tepung sagu, gula dan garam. Kajian ini telah dijalankan ke atas Keropok Lekor untuk melihat kesan kestabilan penyimpanan selama 12 hari dalam keadaan dingin (4C) selepas penambahan tumbuhan tempatan. Formulasi tanpa sebarang penambahan antipengoksida digunakan sebagai sampel kawalan. Formulasi yang lain adalah penambahan campuran herba kering, penambahan herba basah dan juga formulasi dengan penambahan BHA/BHT yang digunakan sebagai perbandingan antipengoksida sintetik dangan antipengoksida tumbuhan semulajadi. Analisis yang telah dijalankan ialah analisis kimia yang dinilai melaui pengiraan hasil pengoksidaan primer (Nilai Peroksida) dan sekunder (Nilai Asid Tiobarbiturik). Penentuan mikrobiologi termasuk analisa Jumlah Kiraan Plat dan analisa Jumlah Kiraan Yis dan Kulat. Ujian penilaian sensori juga telah dijalankan. Herba kering dan basah P. hydropiper dan C. asiatica menunjukkan perencatan pengoksidaan lemak yang signifikan berbanding sampel kawalan. Pada mulanya, Nilai Asid Tiobarbiturik menunjukkan tida perbezaan yang signifikan (P<0.05) dengan Keropok Lekor yang ditambah dengan BHA/BHT. Analisis Jumlah Kiraan Plat menunjukkan kiraan yang tinggi bagi kesemu sampel. Tetapi, sampel rawatan menunjukkan perbezaan signifikan rendah berbanding dengan sampel kawalan sehingga hari ke 9 penyimpanan. Manakala, Kiraan Yis dan Kulat menunjukkan kiraan yang lebih rendah daripada Jumlah Kiraan Plat di mana sampel kawalan menunjukkan perbezaan signifikan tinggi antara kesemua sampel. Penerimaan tertinggi keseluruhan di dalm ujian penilaian sensori adalah Keropok Lekor yang ditambah herba basah. Sebagai kesimpulan, kesan terbaik antipengoksida dan antimikrob adalah daripada Keropok Lekor yand ditambah BHA/BHT. Walaubagaimanapun, penambahan Keropok Lekor dengan herba asli juga mengurangkan pengoksidaan lemak yang menunjukkan kesan antipengoksidaan sehingga hari ke 9 penyimpanan. Kesan antimikrob adalah kurang dalam kesemua sampel yang dirawat.

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CHAPTER 1

INTRODUCTION

1.1

Background and Problem statement Herbs are used in many domains, including medicine, nutrition flavouring, beverages, dyeing, repellents, fragrances, cosmetics. Many species have been recognised to have medicinal properties and beneficial effect on health and food such as antioxidant activity, digestive stimulation action, antiinflammatory, antimicrobial, hypolipedemic, antimutagenic effects and anticarcinogenic potential (Aneta et al., 2007). Traditional herbs of the Malays in Malaysia (locally called ulam) comprise of more than 120 species representing various families, from shrubs to large trees. The leaves, shoots or rhizomes of the vegetable are eaten fresh as salad or cooked. They are consumed because of their taste, which adds variety and flavour to the diet, as well as for health benefit. Nutritional studies have indicated that many of these vegetables are rich in carbohydrate, proteins, minerals and vitamins (Faridah et al., 2006).

Although consumption of fruit and vegetables has increased, it is still lower than in the Mediterranean countries. Therefore the incorporation of seasoning based on herbs into everyday meals may be crucial importance. The benefits resulting from the use of natural product rich in bioactive

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substances has promoted the growing interest of pharmaceutical, food and cosmetic industries as well as of individual consumer in the quality of herbal produce (Ewa et al., 2005). Autoxidation of fats and oils in processed foods maybe prevented by the use of oxidation inhibitors or antioxidants. Plant extracts obtained from some fruits and vegetables have been reported to be effective antioxidants (Vanitha et al., 2005).

Even though many research works have proven the health benefit of herbs, the data collection on the Malay herbs which are known as ulam in Malaysia is still lacking. Furthermore there are no data available on the effect of incorporating the local ulam on the storage stability of traditional food such as Keropok Lekor.

Fish are perishable food commodities, so as fish food products. It generally spoils faster than do other muscle foods (Wenjiao et al., 2008). Keropok Lekor is a product made mainly from fish and is easily spoiled during storage at chilled temperature and normally consumed fresh. Thus it is important to increase the shelf life of this product. The spoilage is a complex processes in which physical, chemical and microbiological mechanism are involved especially bacteriological activity. the deterioration due to lipid oxidation and

1.2

Significance of study This study will be carried out to investigate the effect of Malay herbs such as P. hydropiper (Kesum) and C. asiatica (Pegaga) on the oxidative and the

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antimicrobial stability of Keropok Lekor stored at chilled temperature and therefore increases the shelf life of the product. The herbs are known to contain high antioxidative activities. Malays usually consumed local herbs as salad, but this study will diversify the usage of these local herbs in Keropok Lekor. If the research is successful, a new product with natural preservatives will be obtained. This will be an opportunity for the food industry to commercialise a healthy traditional food in the market.

1.3

Objectives The intention of this research is to investigate the effect of Malay herbs namely Polygonum hydropiper (Kesum) and Centella asiatica (pegaga) on Keropok Lekor. The objectives of this project are: 1. To compare the oxidative and antimicrobial stability of Keropok Lekor incorporated with dry herbal mix (P. hydropiper and C. asiatica), wet herbal mix (P. hydropiper and C. asiatica) and synthetic antioxidant BHA/BHT. 2. To determine the shelf life of the Keropok Lekor incorporated with local herbs. 3. To investigate the acceptability of this product by the consumer.

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CHAPTER 2

LITERATURE REVIEW

2.1

Fish Fish are perhaps one of the most vulnerable of the world resources. For many economically developing nations, fish are the first or second largest export commodity. Since the freshness of fish deteriorates rapidly, freshness may be considered a synonym for quality. The increasing demand of high quality fresh seafood has intensified the search for methods and technologies for better fresh fish utilization (Manju et al., 2007), and also fish product utilisation.

Fish provides a good source of high quality protein and contains many vitamins and minerals. It may be classed as either white (lean), oily (fatty) or shellfish. White fish, such as haddock and seer, contain very little fat (usually less than 1%) whereas oily fish, such as sardines, contain between 10-25%. The latter, as a result of its high fat content, contain a range of fat-soluble vitamins (A, D, E and K) and essential fatty acids, all of which are vital for the healthy functioning of the body. The composition of fish is influenced by feeding, locality, size, age, season and species. Certain processing techniques such as boiling leach the water-soluble vitamins into the surrounding liquid. If this is thrown away, a great deal of nutritional value is lost (Fellows and

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Hampton, 1992). Table 2.1 shows the nutrient composition some of the main nutritional differences between oily and white fish.

Table 2.1 The nutrient composition of oily and white fish

Composition White fish e.g. haddock Oily fish e.g. herring

Energy (KJ) Protein (g) Fat(g) Water (g) Calcium (mg) Iron (mg) Vitamin A (mg) Thiamine (mg)
Source: Fellows and Hampton (1992).

321 17 07 82 16 0.3 0 0.07

970 17 18 64 33 08 45 0

Fish is one of the most highly perishable food products. During handling and storage, quality deterioration of fresh fish rapidly occurs and limits the shelf life of the product. Shelf life is defined as the period of time, under defined condition of storage, for which food products remains safe and fit for use. In other words, during this period, it should retain its desired sensory, chemical, physical, functional or microbiological characteristic (Khalid, 2007). Table 2.2 shows the shelf life of fish products stored at different temperature.

Table 2.2 Predicted shelf life of fish products stored at different temperatures
Shelf life in days of product stored in ice (0C) Shelf life at chill temperatures (days)

5C 2.7 4.4 6.2 8

10C 1.5 2.5 3.5 4.5

15C 1 1.6 2.2 2.9

6 10 14 18
Source: Huss (1995).

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2.1.1 Fish Muscle There are three types of muscles: striated muscles which constitute the livestock meat or fish meat, smooth muscles which constitute mollusks or visceras, and the heart muscle which is more or less an intermediate structure between the other two. Livestock meat and fish meat are basically striated muscles which are formed by group of muscle fibers with striation. Fish muscle can be divided into ordinary muscle (white meat) and dark muscle (dark meat).

Dark muscle lies along the side of the body under the skin. The shape and volume of the dark muscle can be classified in three types according to the fish species (Suzuki, 1981). There are many differences in the chemical composition of the two muscle types, some of the more noteworthy being higher levels of lipids and myoglobin in the dark muscle. From a technological point of view, the high lipid content of dark muscle is important because of problems with rancidity (Huss, 1995).

The muscle fiber lies parallel to one another within myotome, divided by the membrane consisting of the mycommata. The myotome of the back and belly are arranged in concentric circles. Muscle fiber, smaller unit of muscle, are bound together by connective tissue (endomysium) and covered with mycommata. Muscle fiber consists of many myofibril which lie parallel, and sarcoplasm which fills the space. There is more connective tissue around the muscle fibers in dark muscle than in ordinary muscle meat (Suzuki, 1981).

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2.1.2 Shortfin Scad Fish Scad belong to the family Carangidae. They resemble mackerel in flavor, but a little milder and without so much oil. Shortfin Scad or Round Scad (Decapterus macrosoma) is an Indo-Pacific scad and is also found in the East Pacific from the coast of Baja California to Northern Peru. They can grow to over 14 inches, but the ones popular are about 6-1/2 inches and weigh 2 ounces (http://www.mexfish.com). In Malaysia, Shortfin scad is known as Selayang, Curut, Sardin or Basung (Ismail et al., 2004). This fish is excellent for frying whole. It has a strong enough flavor to be interesting hot or cold and, while the head and spine are too hard to eat, it has no bone or fin problems otherwise. Table 2.3 shows the nutrient composition of Shortfin scad fish.

Table 2.3 Nutrient composition of Shortfin scad fish

Type Weight (g) En (Kcal) Water (g) Nutrient composition Protein (g) Fat (g) CHO (g) Fiber (g) Ash (g)

1 piece, tail portion 1 whole, small 1 whole medium

48

43

28.6

8.5

0.9

0.0

0.0

0.6

81

41

27.7

8.3

0.9

0.0

0.0

0.6

105

54

36.2

10.8

1.2

0.0

0.0

0.7

Source: Tee, et al. (1997).

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2.1.2.1 Keropok Lekor Keropok lekor is a popular traditional Malay snack food in the East Coast Malaysia which has a resemblance to a sausage in appearance. They are marketed in variable sizes such as those with a diameter of 4.0-6.0 cm and 1.5 2.0 cm. The bigger size Keropok Lekor are normally sliced to a thickness of 1.0 1.5 cm or to a thickness of 2.0 2.5 mm prior to deep frying. The smaller ones are normally slit and fried whole. The main characteristics of fried Keropok Lekor are the crunchiness on the surface and chewiness in the inner part of the product.

The two essential ingredients in Keropok Lekor making are starch and fish. Fish such as Ikan Parang (Chirocentrus dorab), Ikan Tamban beluru (Clupea leiogaster) and Ikan Selayang (Decapterus macrosoma) are preferred although other fishes are also used for making Keropok Lekor. Tapioca or sago starch is used but sago starch is said to give the best product in terms of texture and flavour (Muhammad Redza, 2004). Every food products final profile will be established by the ingredients and processing (Modi et al., 2006).

This food product can be eaten as soon as it is boiled and together with chili sauce. Keropok Lekor usually has a short shelf life. When stored in the fridge especially at chilled temperature the Keropok Lekor does not last long. After a few days, it will have a slimy layer and people usually just wash the slimy layer and add a bit of salt then boiled and then fry it. As it is known the slimy layer is due to bacteriological deterioration.

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2.1.3 Chilled fish The appearance of fish stored under chilled condition without ice does not change as much as for iced fish, but the fish spoil more rapidly and an evaluation of cooked flavour will be necessary. Temperature for chilling is usually between 0-25C. It is well known that both enzymatic and microbiological activities are greatly influenced by temperature. However, in the temperature range from 0 to 25C, microbiological activity is relatively more important, and temperature changes have greater impact on microbiological growth than on enzymatic activity. Many bacteria are unable to grow at temperatures below 10C and even psychrotrophic organisms grow very slowly, and sometimes with extended lag phases, when temperatures approach 0C.

Several studies have been carried out on the quality changes of ready-made seafood products. One of them was performed by Akkus et al. (2004) on fish balls that were preapared using boiled and raw anchovy. These authors discovered that the shelf-life of the fish balls was 9 days at 41C. Gkoglu (1994) used sensory, physical, chemical and, and microbiological analysis to study fish balls made from mackerel that were prepared using boiling (precooking) method and stored at a refrigerated temperature. In this study, a shelf-life of up to 8 days and spoilage after 10 days were reported for fish balls stored under refrigerated temperature.

The microflora responsible for spoilage of fresh fish changes with changes in storage temperature. At low temperatures (0-5C), Shewanella putrefaciens,

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Photobacterium phosphoreum, Aeromonas spp. and Pseudomonas spp. cause spoilage. However, at high storage temperatures (15-30C) different species of Vibrionaceae, Enterobacteriaceae and Gram-positive organisms are responsible for spoilage. For tropical fish, however, the average relative rate of spoilage of a large number of species stored at 20-30C was approximately 25 times higher than at 0C (Huss, 1995).

Minced fish is commonly used for product of surimi-based or traditional, ready-to-eat, pre-cooked products such as fish balls or burger patties. Three types of fish mince from whiting (Merlangius euxinus) which is plain mince commonly used in Turkey for producing fish balls and requires no washing steps, surimi commonly used for making ready-made products and the type produced from boiled fish which is used in household preparation of fish balls. All mince was stored under refrigerated conditions (at 41 C) for 15 days. Increasing amounts of TVB-N, TMA-N and TBA were found in all types of mince with increasing storage time. The results of measured chemical parameter, plain mince retained its consumable properties up to 11 days of storage; surimi was acceptable for consuming up to 13 days of storage. Pre-cooked product was still consumable up to the end storage time. However, sensory attributes showed only 7 days of storage life for all types of products (Sevim et al., 2006).

If quality criteria of chilled fish during storing are needed, sensory assessment of the cooked fish can be conducted. A characteristic pattern of the deterioration of fish stored in ice can be found and divided into the following

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four phases, in phase 1 the fish is very fresh and has a sweet, seaweedy and delicate taste. The taste can be very slightly metallic. In cod, haddock, whiting and flounder, the sweet taste is maximized 2-3 days after catching. For phase 2 there is a loss of the characteristic odour and taste. The flesh becomes neutral but has no off-flavour. The texture is still pleasant. As for phase 3 there is sign of spoilage and a range of volatile, unpleasant-smelling substances is produced depending on the fish species and type of spoilage (aerobic, anaerobic). One of the volatile compounds may be trimethylamine (TMA) derived from the bacterial reduction of trimethyl-aminoxide (TMAO). TMA has a very characteristic "fishy" smell. At the beginning of the phase the off-flavour may be slightly sour, fruity and slightly bitter, especially in fatty fish. During the later stages sickly sweet, cabbage-like, ammonia, sulphurous and rancid smells develop. The texture becomes either soft and watery or tough and dry. And at phase 4 the fish can be characterized as spoiled and putrid (Huss, 1995).

2.2

Spoilage in Fish Fish is an extremely perishable food. For example, most fish become inedible within twelve hours at tropical temperatures. Spoilage begins as soon as the fish dies, and processing should therefore be done quickly to prevent the growth of spoilage bacteria. Fish is a low acid food and is therefore very susceptible to the growth of food poisoning bacteria. This is another reason why it should be processed quickly. Some methods of preservation cause changes to the flavour and texture of the fish which result in a range of different products. These include cooking (for example, boiling or frying),

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lowering the moisture content (by salting, smoking and drying collectively known as curing), lowering the pH (by fermentation) and lowering the temperature with the use of ice or refrigeration also preserves the fish, but causes no noticeable changes to the texture and flavour (Fellows and Hampton, 1992).

Fish are perishable food commodities, which generally spoil faster than do other muscle foods. Deterioration of fish mainly occurs as a result of bacteriological activity leading to loss of quality and subsequent spoilage. The spoilage of fish is a complex process in which physical, chemical and microbiological mechanisms are implicated (Wenjiao et al., 2008). Fish tissue is characterized in being rich in protein and non-protein nitrogen (e.g. amino acids, trimethylamine-oxide (TMAO), creatinine, but low in carbohydrate resulting in a high post mortem pH <6.0. Further, the pelagic, fatty fishes have a high content of lipids consisting mainly of triglycerides with longchain fatty acids which are highly unsaturated. Also the phospholipids are highly unsaturated and these circumstances have important consequences for spoilage processes under aerobic storage conditions (Huss, 1993).

Obvious signs of spoilage are: detection of off-odours and off-flavours, slime formation, gas production, discolouration, changes in texture and the development of these spoilage conditions in fish and fish products is due to a combination of microbiological, chemical and autolytic phenomena (Huss, 1993). Fish are also prone to oxidation, resulting in off flavour, changes in colour and texture and nutrients (Rabia et al., 2008). The onset of rancidity is

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faster particularly in fatty and semi-fatty species like horse mackerel, in whose muscle has large amount of hemoglobin a well known activator of lipid oxidation and lipid coexist (Snchez-Alonso et al., 2008). Table 2.4 shows the causes of the various types of spoilage.

Table 2.4 The causes of the various types of spoilage in fish

Causes of fish spoilage Signs of spoilage Microbiological Chemical (oxidation) Autolytic Physical

Off odours/ off flavours Slime formation Gas formation Discolouration Change of texture
Source: Huss (1995).

+ + + + +

+ + -

+ + +

+ +

2.2.1 Microbiological spoilage Initial loss of quality of fresh lean or fatty fish species, chilled or not chilled, is caused by autolytic changes, while spoilage is mainly due to the action of bacteria. During storage a characteristic flora develops, but only a part of this flora contribute to spoilage. The specific spoilage organisms (SSO) are producers of the metabolites responsible for the off odours and off flavours associated with spoilage.

Shewanella putrefaciens is typical for the aerobic chill spoilage of many fish from temperate waters and produces trimethylamine (TMA), hydrogen

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