Microbiological quality analysis of freshwater prawns from three sampling sites in Peninsular Malaysia viz: Site 1- Kg. Jumbang, Negri Sembilan; Site 2- Kg. Cangkat Tin, Perak and Site 3- Kg. Cenderiang, Perak for total mesophilic and psychrophilic aerobic counts, proteolytic bacterial counts, histamine producing bacteria, cadaverine producing bacteria and putrescine producing bacteria in the prawns and pond water for the three sites showed that the microbiological quality of freshwater prawns is related to the microflora of pond water in which they were grown. The initial bacterial counts indicated the values were in the range of log 4+ CFU/g for all samples. Total mesophilic and psychrophilic counts of the head regions were higher than that of the body regions for all prawn samples and types of growth media tested. All samples showed an increase in counts with time and temperature of storage up to log 7+ CFU/g for mesophilic counts after 12 hours at ambient, 6 days at 10 ± 2°C and 12 days at iced storage. The samples from Site 2 had relatively higher counts compared to the other two sites which correlated well with the levels determined in the pond water. Similar trends were observed for psychrophilic counts but at lower values for the different types of media studied.
Effects of preservatives on quality changes and shelf life of shrimp during iced storage indicated that boric acid, lactic acid and sodium metabisulphite managed to inhibit psychrophilic bacteria and biogenic amines formation in prawns while maintaining the mesophilic counts at lower levels during iced storage.
Biochemical analysis was carried out for pH profiles, freshness in terms of K-values, amino acids profiles, total volatile bases (TVB), total volatile acids (TVA) and biogenic amines for fresh and preserved Macrobrachium rosenbergii. Results showed that pH profiles of Macrobrachium rosenbergii explain the inability of this parameter to be used to evaluate the quality of Macrobrachium rosenbergii. Thus changes in pH profiles of Macrobrachium rosenbergii should be combined with indicators such as total volatile acids and total volatile bases. Total volatile acids of the shrimps increased steadily in small amounts throughout the storage period. A rapid increase in TVB at 100C was detected due to the increase in total aerobic bacteria at elevated temperatures. The microbial activities caused the decrease in the amino acids arginine, lysine and histidine which correlated well with the increase in the corresponding biogenic amines such as putrescine, cadaverine and histamine respectively. Preservatives used in this study controlled the production of these biogenic amines without significantly altering the pH of preserved shrimp.
Vibrio parahaemolyticus is prevalent in tropical marine environment in all seasons and can cause seafood-borne gastroenteritis. A total of 251 suspected isolates were tested including 60 from frozen shrimp, 50 from cultured live shrimp, 67 from sediments of culture ponds and 74 from water were subjected to polymerase chain reaction (PCR) targeting the toxR gene for confirmation as V. parahaemolyticus. Of the 128 toxR positive isolates, 15% of the isolates from culture environment (from live shrimp, sediments and water) and 7% of frozen shrimp samples were positive for the tdh and trh genes. Since urease production could be a marker of trh but not tdh in V. parahaemolyticus, a total of 189 of the 251 suspected V. parahaemolyticus isolates were tested for urease production and 41% of the isolates were found to be positive for urease production. However not all urease positive strains of V. parahaemolyticus were positive for either tdh or trh genes. Detection of virulent strains in shrimp culture environment in Malaysia suggests a probable risk for health of people consuming raw shrimp.
The concentrations of cadmium, copper, zinc and lead, in the total soft tissues of green-lipped mussel Perna viridis of a wide range of sizes (2-11 cm), were determined from a population at Pasir Panjang. The metal contents (μg per individual) and concentrations (μg per g) of cadmium, lead, copper and zinc were studied in P. viridis to find the relationships with body sizes. Smaller and younger mussels showed higher concentrations (μg per g) of Cd, Pb and Zn than the larger and older ones. The results of the present study showed that the plotting of the metal content, against dry body flesh weight on a double logarithmic basis, gave good positive straight lines; this observation is in agreement with Boyden’s formula (1977). This indicated that P. viridis showed a different physiological strategy for each metal being studied, which is related to age.
Hepatitis A virus infection occurs globally and is causing a public health concern, primarily in developing countries due to its persistent circulation in the environment. The improved sanitary condition and increase in awareness of personal hygiene have led to the marked reduction of HAV prevalence in industrialized countries during childhood and to a shift of the infection towards adulthood. HAV is an environmentally stable, positive single stranded RNA virus that is primarily transmitted by the fecal-oral route, person to person contact or ingestion of contaminated food and drink. One of the main causes leading to HAV infection is epidemiologically linked to the consumption of raw or undercooked shellfish particularly oysters and clams. Due to their filter-feeding style, these shellfishes readily concentrate viruses from the surrounding water containing municipal sewage, and as a consequence pose a health threat to consumers. Therefore, development of detection techniques possessing the requisite sensitivity and specificity for the practical routine monitoring purposes is of great importance necessary for the protection of shellfish-consuming public. Nucleic acid based method such as reverse transcription PCR has emerged as the popular method of choice in view of its rapidity, accuracy and
sensitivity in contrary of the time-consuming conventional cell culture and hybridization techniques. However, detection of hepatitis A virus is firstly hampered by the non-cytophatic effect of wild type HAV strain, secondly, the low concentration of viral genome present in the environmental sample which requires effective isolation and concentration of virions and lastly the labor-extensive purification and thorough removal of the abundance of the PCR inhibitors which will unfavorably reduce the efficiency of PCR detection.
Vibrio parahaemolyticus is one of the most widely recognized pathogenic Vibrio species due to numerous outbreaks and its’ wide occurrence in marine environment. In this study, 32 isolates of V. parahaemolyticus isolated from cockles were tested for sensitivity to 16 antibiotics and the presence of plasmids. All the isolates were multi-resistance, defined as resistant to atleast three different antibiotics with multiple antibiotic resistance indexes ranging from 0.31 to 0.69, indicating the isolates originate from high risk sources of contamination where antibiotics are often used. In the plasmid profiling test, only 15 isolates (47%) harbored plasmid DNA, which ranged in size from 2.7 to 56.2 kb, separating the isolates into 14 plasmid profiles. Hence, food contaminated with antibiotic resistant V. parahaemolyticus could be a major threat to public health due to the distinct possibility that they can be a significant reservoir of genes encoding antibiotic resistance determinants that can be transferred intra or interspecies. As in many developing countries, raw food hygiene and antimicrobial resistance epidemiology is still in the infancy stage in the locality of the study and thus our data provide a current baseline profile of antimicrobial resistance and plasmid of V. parahaemolyticusfrom cockles in Padang, Indonesia.
Allergy to different classes of mollusks, including squid, which are members of the class Cephalopods has been reported. Tropomyosin, a major muscle protein, is the only well-recognized allergen in squid. The aim of this study was to characterize IgE-binding proteins of local Loligo edulis (white squid) consumed in Malaysia. Protein profiles and IgE-binding proteins were detected by sodium dodecyl sulfate-polyacrylamide gel-electrophoresis (SDS-PAGE) and immunoblotting using sera from 23 patients with positive skin prick test to raw squid extract. SDS-PAGE of the raw extract exhibited 21 protein bands (10-170 kDa) but those ranging from 19 to 29 kDa and 41 to 94 kDa were not found in the cooked extract. Immunoblotting of raw extract demonstrated 16 IgE-binding bands, ranging from 13 to 170 kDa. A heat-resistant 36 kDa protein, corresponding to squid tropomyosin, was identified as the major allergen of both extracts. In addition, a 50 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. Our findings indicate that the allergen extract used for diagnosis of squid allergy should contain both the 36 kDa and 50 kDa proteins.
The shrimp paste called belacan is a traditional umami taste condiment extensively used in Malaysia that is rich in glutamate and 5'-nucleotides. The aim of this study was to determine the concentration of glutamate and 5'-nucleotides of various types of foods prepared with belacan and to measure their sensory attributes. The concentration of free glutamic acid found in different brands of belacan was 180-530mg/100g and in local dishes 601-4207mg/100g. The total amount of 5'-nucleotides in belacan samples ranged from 0.85 to 42.25μg/g. A Quantitative Descriptive Analysis (QDA) using a list of 17 sensory attributes showed a good correlation between belacan concentration in the final food and a range of positive sensory attributes, except for bitter, sweet, sour taste and astringency. Belacan also contains bitter, sweet and sour compounds that change the positive attributes of belacan at higher concentrations. The highest aroma attributes were linked to nasi goreng belacan (belacan fried rice) while the highest flavour attributes were found in sambal belacan. There was a 32 folds significant increase of umami attributes with the addition of belacan to final foods. The optimum amount of belacan was 0.45% for asam pedas (tamarind flavoured dish with belacan), 18% for sambal belacan (chilli belacan), 1.5-2.5% for kangkong goreng belacan (stir fried water convolous with belacan), and 2% for nasi goreng belacan.
Aimed of this study was to determine the presence of Vibrio cholerae in cockles (Anadara granosa)
from different coasts in Malaysia and to measure the biosafety of V. cholerae in raw cockles at wet market in Malaysia using the polymerase chain reaction (PCR) in combination with the most probable number (MPN) method. A total of 100 samples from 4 different wet markets in the West and East were examined for the presence of V. cholerae. The prevalence of V. cholerae between the two coasts was not significant different. In fact, the 74% of samples from West coast area was found positive while the 69% for samples collected in the East coast. West coast samples showed a prevalence of 60% for the wet market A=, 64% for B=, 88% for C= and 84% for the market D); East coast samples showed the same percentage with 72% for the wet markets E, F and H, followed by wet market G with 60%.With the MPN-PCR method, using 80 samples of raw cockles obtained from 4 wet markets, the occurrence of V. cholerae detected was of 95%. The frequency of V. cholerae in raw cockles obtained from wet market I and L was higher (100%) compared to other wet market (Wet market B=, 90%; Wet market C=, 95%).The density of V. cholerae detected in all samples ranged from 24000 MPN/g, but most of the samples (24 samples) were in category >24000 MPN/g concentration. V. cholerae was present in raw cockles in higher number. Hence, these results demonstrate the presence of pathogenic V.cholerae in cockles harvested and reveal the potential risk of illness associated with their consumption. This study will be the first biosafety assessment of V. choleare in raw cockles in Malaysia and it will provide significant insights about Malaysian scenario.
This study was conducted to improve the formulation of chili shrimp paste (CSP) based on sensory evaluation in terms of acidity, source of acid and coarseness of chili paste and to improve the production process of CSP. The effectiveness of dimethyl dicarbonate (DMDC) as a microbial reduction agent was also evaluated. To produce CSP with different coarseness, a milling machine was used. Two types of preference test were conducted, i.e. ranking and hedonic. The preferred pH level was 4.0, the best acid source was kalamansi juice, and the most preferred coarseness for chili paste was when milling plates with a gap of 120 µm was used. DMDC has no effect on microbial reduction due to the presence of fat globules in CSP which hindered the inactivation action. Milling can substitute pounding as it is much faster and can produce a uniform CSP with higher volume.
This study was conducted to determine the proximate composition and energetic value of selected marine fish and shellfish from the West Coast of Peninsular Malaysia. This study has included 20 species of fish (10 pelagic fish and 10 demersal fish) and 4 species of shellfish. For pelagic fish, Longtail shad (terubuk) was found to contain significantly lower moisture (59.31+0.00%), but significantly higher fat content (23.15+0.00%) compared to other fish species. For demersal fish, moisture and ash contents ranged between 74-82%, and 0.9-2.1%, respectively. Long-tailed butterfly ray contained the highest protein (22.22+1.24%) compared to other fish studied. The highest fat content of demersal fish was in Moonfish at 6.89+2.76%. For shellfish, prawn contained the highest protein (19.12+1.44%). The fat content of shellfish ranged between 1-2%. Oyster contained significantly higher carbohydrate at 6.45+0.00%, compared to other shellfish. Longtail shad contained the highest energetic value of 13.34 kJ g-1 of all samples. These values are useful references for consumers in order to choose fish and shellfish based on their nutritional contents.
A total of sixty V. parahaemolyticus strains isolated from local cockles (Anadara granosa) were investigated by their antibiotic resistance patterns and plasmid profiles. The isolates showed multiple resistances towards most of the antibiotics tested. All strains of V. parahaemolyticus isolated harbored 1-3 plasmids, with sizes ranging from 2.7 to 54 kb. All V. parahaemolyticus strains showed high multiple antibiotics in frequencies of 0.58 – 0.94 indicating that the strains were derived from high-risk sources. In addition, no particular plasmid profile was predictive of a particular pattern of antibiotic susceptibility. These findings are essential because of the suggested involvement of seafood especially shellfish and environment in transmission of this pathogen to human. Thus, indicating that seafood may be a source of food- acquired antibiotic resistant bacteria to consumer.
Hepatitis A is a liver infection caused by the hepatitis A virus (HAV). Outbreaks of hepatitis A have been linked to the consumption of both raw and cooked shellfish. These outbreaks could induce a public confidence problem over shellfish safety and may result in important economic losses for the seafood industry. The work presented in this study investigated the presence of HAV in shellfish from Peninsular Malaysia. A total of 365 of cultured and wild shellfish from 36 sampling locations located throughout Peninsular Malaysia were examined using a commercial nucleic acid extraction and reverse transcription -polymerase chain reaction (RT-PCR) kit. HAV was not detected in almost all of the shellfish samples xamined. Only one cockle sample from Changkat, Seberang Perai was positive for HAV. The results suggest the absence of HAV or very low amount of HAV viral particles in most of the shellfish examined.
This study evaluates the sanitary and physico-chemical quality of Sg. Jarum Mas shellfish waters in order to establish its suitability as a model farm. Seawater and shellfish from nine stations (4 shellfish harvesting waters, 4 surrounding waters and 1 control site) were collected and analyzed monthly from September 2004 - September 2005. The results show that shellfish harvesting waters in Sg. Jarum Mas can be classified as ‘approved’ and ‘conditionally approved’. Hepatitis A virus was not detected in any of the shellfish examined. Dinophysis caudata and Pseuodonitzshia spp. were the most common harmful alga species observed. Harmful species that are known to produce toxins and cause shellfish poisoning such as Alexandrium spp., Gymnodinum spp., Pyrodinium sp. and Prorocentrum spp. were not detected. The physico-chemical characteristics of shellfish waters in Sg. Jarum Mas imply that they are suitable for aquaculture activity of moderately tolerant species such as shellfish.
The use of simple crude water extracts of common herbs to reduce bacterial attachment may be a cost-effective way to control bacterial foodborne pathogens, particularly in developing countries. The ability of water extracts of three common Malaysian herbs (Andrographis paniculata, Eurycoma longifolia, and Garcinia atroviridis) to modulate hydrophobicity and attachment to surfaces of five food-related bacterial strains (Bacillus cereus ATCC 14576, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 10145, Salmonella Enteritidis ATCC 13076, Staphylococcus aureus ATCC 25923) were determined. The bacterial attachment to hydrocarbon assay was used to determine bacterial hydrophobicity. Staining and direct microscopic counts were used to determine attachment of bacteria to glass and stainless steel. Plating on selective media was used to determine attachment of bacteria to shrimp. All extracts were capable of either significantly ( P < 0.05) increasing or decreasing bacterial surface hydrophobicity, depending on the herb extract and bacteria combination. Bacterial attachment to all surfaces was either significantly (P < 0.05) increased or decreased, depending on the herb extract and bacteria combination. Overall, hydrophobicity did not show a significant correlation (P > 0.05) to bacterial attachment. For specific combinations of bacteria, surface material, and plant extract, significant correlations (R > 0.80) between hydrophobicity and attachment were observed. The highest of these was observed for S. aureus attachment to stainless steel and glass after treatment with the E. longifolia extract (R = 0.99, P < 0.01). The crude water herb extracts in this study were shown to have the potential to modulate specific bacterial and surface interactions and may, with further work, be useful for the simple and practical control of foodborne pathogens.
Crab is an important source of food allergen. Tropomyosin represents the main crab allergen and is responsible for IgE cross-reactivity between various species of crustaceans. Recently, other new crab allergens including arginine kinase have been identified. However, information on allergens of the local Portunidcrab is not available. Thus, the aim of this study was to identify the major allergens of Portunus pelagicus (blue swimming crab) using the allergenomics approach. Raw and cooked extracts of the crab were prepared from the crab meat. Protein profile and IgE binding pattern were demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from 30 patients with crab allergy. The major allergens of the crab were then identified by two-dimensional electrophoresis (2-DE), followed by mass spectrometry analysis of the peptide digests. The SDS-PAGE of raw extract revealed approximately 20 protein fractions over a wide molecular weight range, while cooked extract demonstrated fewer protein bands. The raw extract also demonstrated a higher number of IgE reactive bands than the cooked extract. A heat-resistant protein of 36 kDa has been identified as the major allergen in both raw and cooked extracts. In addition, a heat-sensitive protein of 41 kDa was also recognized as a major allergen in raw crab. The 2-DE gel profile of the raw extract demonstrated about >100 distinct proteins spots and immunoblotting of the 2-DE profile demonstrated at least 12 different major IgE reactive spots with molecular masses between 13 to 250 kDa and isoelectric point (pI) values ranging from 4.0 to 7.0. The 36 and 41 kDa proteins were identified as the crab tropomyosin and arginine kinase, respectively by mass spectrometry. Therefore, this study confirmed that tropomyosin and arginine kinase are the major allergens of the local Portunid crab, P. pelagicus.
Vibrio cholerae still represents a significant threat to human health worldwide despite the advances in hygiene, consumer knowledge, food treatment and food processing. In Malaysia, statistics in year 2009 have shown that among the food and water borne diseases, food poisoning has the highest incidence rate of 36.17 per 100,000 populations and with a mortality rate of 0.01 per 100,000 populations. In this study, 22 seafood samples comprising of fish, squid, crustacean and mollusks purchased from wet market and supermarket were analyzed. The Most Probable Number (MPN) and real time PCR was used to enumerate the Vibrio cholerae in seafood sample. The results showed that MPN-real time PCR of the samples from wet market had a maximum of >1100 MPN/g compare to 93 MPN/g enumerated from the MPN plate. The MPN-real time PCR in the samples from supermarket indicated 290 MPN/g as compared to 240 MPN/g enumerated from the MPN plate. The standard curves showed that there was a good linear correlation between the Ct values. The minimum level of detection of Vibrio cholerae standard DNA at targeted gene was 3 x 10-5 ng/μl.
Level of heavy metals in marine ecosystem has been intensively studied during recent years as these hazardous substances could be accumulated in the biota. Generally, the presence of contaminants in fish is a result of human activities such as industrial and agricultural wastes. In this study, the marine fish and shellfish from the Straits of Malacca were analyzed using Inductively Coupled Plasma Optical Emission Spectrophotometer (ICP-OES) and Flow Injection Mercury System (FIMS) for Cd, As and Pb and Hg, respectively. The Straits of Malacca is one of the busiest shipping routes in the world that make the level of heavy metals potentially high besides the various industrial activities along the west region of Peninsular Malaysia. The range of heavy metals in samples were 1.0-3-6.5-3 µg/ g wet sample for Hg, 0.5-2-47-2 µg/ g wet sample for Cd, 0.01-0.39 µg/ g wet sample for Pb and 0.14-6.57 µg/ g wet sample for As. Most part of the values was below the permitted limit set by FAO/WHO 2004 as well as Food Act 1983 and Food Regulations 1985. Therefore it can be generalized that fish and shellfish from the Straits of Malacca are safe to consume in terms of these heavy metals concentrationand do not constitute a risk for human health.
The total lipid contents and the concentration of eicosapentaenoic (EPA) and docosahexaenoic
(DHA) fatty acid of fourteen selected marine, three freshwater fish species, four shellfish species and two selected canned fish species of East Coast of Peninsular Malaysia were determined. The fat content of all samples ranged from 1.01 % to 15.83 % with silver catfish reported to have the highest value. Most of the fish had fat amounts lower than 10% of their total weight. In general, DHA concentrations (50.50-165.21 µg/g) were significantly higher than EPA (11.12-55.38 µg/g) in all of the fish species analyzed. Among all marine fish species, the sixbar grouper recorded the highest concentration of DHA (165.21 µg/g) while barramundi had the highest concentration of EPA (55.38 µg/g). In conclusion, all fish and shellfish species are considered a good source of EPA and DHA, representing a very valuable essential nutrient for maintenance of human health.
Harmful algal blooms (HABs) events have been increasingly reported in the country, not only of the frequency and severity of the events, but also involved more species than previously known. In this paper, a decadal review of HABs events in Malaysia is summarized. Bloom events caused by harmful dinoflagellate species including the shellfish poisoning events were highlighted. Paralytic shellfish poisoning (PSP) is no longer restricted to Sabah coasts and Pyrodnium bahamense. Bloom of Alexandrium minutum was reported for the first time in the Peninsula with six persons hospitalized including one casualty after consuming the contaminated benthic clams. Algal blooms that are associated with incidence of massive fish kills have been reported from both east and west coasts of the Peninsula in conjunction to finfish mariculture loses. The culprits of these bloom events have been identified as the dinoflagellates, Cochlodinium polykrikoides, Neoceratium furca, Prorocentrum minimum, Noctiluca scintillans and a raphidophyte, Chatonella ovata. In this paper, some of these HABs species were characterized morphologically and genetically, including their toxicity. Therefore, with the increase of coastal utilization and eutrophication, prevention, management and mitigation strategies, such as site selection, moving pens, clay spraying should be adopted to minimize the impact of these natural events.