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.
The status ofhve l1eavy metals: cadmium, lead copper, zinc and mercury were determined in seafood and its products imported_ti·om Thailand via Bukit Kayu Hiram, Kedah, Flame Atomic Absorption Spectrophotometer was used to determine the level of these heary metals except for mercury, where the cold vapor technique was used, Randomized sampling was done according to a predetermined sampling plan based on the previous years consignments. Data collected were compared with the maximum permitted level of -metal contaminants in fish and fish products- ofthe Fourteenth Schedule (Regulation 38) of Food Act I983 and Food Regulations I 985 to ascertain compliance. lt was noted form this study, that the level of metals detected in seafood and its products had a very wide range, The levels detected for cadmium was at a range of 0. 00] - 3.9/2, 0.07 — 0.29, 0,04 - 4,4 mg/kg in fish, shellfish and cuttlefish respectivelv. In general, cadmium level in some samples was notably higher particularly in shellfish. All samples had lead level less than the permitted value except for fish where the highest value detected was 3.28 mg/kg. The level of copper and zinc was higher than the permitted value in octopus, prawn and crab, Mercury level in all samples analvzed was found to be below the detection level. As for fish samples, zinc level was found to be higher whereas copper was within the limit. However, it was also noted that the level of all the heavy metals in jiozen jish was within the permitted limit.
Biological amines are nitrogenous compounds that occur naturally in wide variety of food. Histamine, putrescine, cadavarine, tyramine, spermine, spermidine, tryptamine and β-phenylethylamine are the biogenic amines that are normally present in foods. Although the biogenic amines play some important physiological functions but high level of amines can cause toxicological effects. High amount of amines can be produced by bacteria during amino acids decarboxylation and have been identified as one of the important agent causing seafood intoxication. Temperature is the major factor for controlling the biogenic amines formation in food. The effects of other alternatives are also discussed including salting, packaging, irradiation, high pressure processing and the use of starter culture. A variety of techniques can be combined together to control the microbial growth and enzyme activity during processing and storage for better shelf life extension and food safety.
Antibiotic susceptibility and genetic diversity of E. coli isolated from cultured catfish and their surrounding environment were determined. The levels of resistance of the E. coli isolates towards six different antibiotics tested differed considerably. Though the isolates displayed resistance towards some of the antibiotics tested, none of the isolates showed resistant towards norfloxacin, sulphametoxazole/trimethoprim and chloramphenicol. RAPD-PCR analysis using single primer and primers combination clustered the E. coli isolates into 3 and 5 groups, respectively. The results of this study suggest that the E. coli isolates from the catfish and their surrounding environment derived from a mixture of sensitive and resistant strains with diverse genetic contents. The use of the RAPD analysis is sufficiently discriminatory for the typing of the E. coli isolates.
Gelatin from fish skin is known to be an alternative source for mammalian gelatin. However, it has weaker properties compared to bovine and porcine gelatin, which limits its use in the industry. The conventional method for fish gelatin extraction requires long production time and could cause serious water pollution and chemical treatments are often being used to enhance the yield of fish gelatin and its properties but it may affect the amino acid content of the gelatin. In this regard, High-Pressure Processing (HPP) is a novel method suggested for fish gelatin extraction. The HPP method is classified as green technology as it requires low electricity throughout the process. This study will discuss the impact of HPP the technique gelatin extracted from fish skin. Skins from four types of fish, namely red tilapia (Oreochromis niloticus), black tilapia (Oreochromis mossambicus), grouper (Epinephelus areolatus) and threadfin bream (Nemipterus tambuloides), were used. High pressure was applied at either pretreatment in citric acid solution or during thermal extraction; and the pressure was maintained at 250 MPa with pressure holding time of 10 minutes and 18 hours of water extraction. Gelatin extract from traditional acid-base method was prepared as a standard for comparison. The study found that there was an increment in the yield of gelatin and the concentration of gelatin extract, and the pre-treatment time was also reduced.
The current study was conducted to evaluate the nutritional characteristics (moisture, protein, lipids, ash and fatty acid composition) of the flesh of oil sardine (Sardinella longiceps) and Indian mackerel (Rastrelliger kanagurta) caught from Hadhramout coast of the Arabian Sea. The protein content was 21.6% and 18.1% (wet weight basis) for mackerel and sardine, respectively. The lipid content was much higher in sardine (10.1%) compared with mackerel (1.7%). The fatty acid composition showed that total saturated fatty acids had the highest relative value (37.5%) among other fatty acid groups in the flesh lipids of sardine, followed by polyunsaturated fatty acids (29.9%) and monounsaturated fatty acids (23.4%). In mackerel, polyunsaturated fatty acids was present at 37.4%, followed by saturated fatty acids (36.7%) and then monounsaturated fatty acids (14.3%). The majority of polyunsaturated fatty acids in both fish were deposited as omega-3 (89.8% in sardine and 87.9% in mackerel), of which docosahexaenoic acid and eicosapentaenoic acid were the most abundant. In conclusion, oil sardine and Indian mackerel which are locally available and affordable fish in Yemen can be considered valuable sources of nutrients particularly protein and health-beneficial omega-3 long chain polyunsaturated fatty acids.
This article contains data on the bacterial communities and its diversity associated with Anadara granosa. The A. granosa samples were obtained from two major estuaries in Penang, Malaysia using a culture dependent and 16S rRNA Illumina sequencing approaches. A. granosa, a commercial blood cockles and popular seafoods, is fragile to the surrounding environments. Thus, our research focused to better understand the bacterial communities and it diversity in the A. granosa, as well as on the generation of a metagenomic library from A. granosa to further understanding on it diversity. The bacteria Vibrionaceae (34.1%) was predominant in the A. granosa from both environments followed by Enterobacteriaceae (33.3%) and Bacillaceae (16.75%). Vibrio sp., Klebsiella sp., and Bacillus subtilis were the most abundant species present. The data generated in this research is the first metagenomic examination of A. granosa and will provide as a baseline to understand the bacterial communities associated with A. granosa and its surrounding natural environments.
This study was conducted to determine the cholesterol and alpha-tocopherol contents of 20 marine fish and four other seafood from the Straits of Malacca. Cholesterol and alphatocopherol contents of the fish and other seafood were determined using high-performance liquid chromatography. The results showed that most of the fish contained low amounts of cholesterol, except sixbar grouper (Epinephelus fasciatus), long-tailed butterfly ray (Gymnura sp.), yellowstripe scad (Selaroides leptolepis), cuttlefish (Sepia officinalis), large-scale tongue sole (Cynoglossus arel), and longtail shad (Hilsa macrura) that contained high amounts of cholesterol (119.39-353.97 mg/100 g wet samples). Indian mackerel (Rastrelliger kanagurta), giant seaperch (Lates calcarifer), prawn (Metapenaeus affinis), and moonfish (Trachinotus blochii) had high alpha-tocopherol contents (462-989 μg/100 g wet sample). Regular consumption of fish and other seafood is highly recommended partly due to the high alphatocopherol content. Due to the high cholesterol in certain types of fish, consumption of the fish fillets of sixbar grouper, long-tailed butterfly ray, yellowstripe scad, cuttlefish, and large scale tongue sole should be < 100 g per day and < 50 per day for longtail shad. Validation of the analytical method also showed a high accuracy and reproducibility of the HPLC method.
Aquaculture production of the Pacific white shrimp is the largest in the world for crustacean species. Crucial to the sustainable global production of this important seafood species is a fundamental understanding of the shrimp gut microbiota and its relationship to the microbial ecology of shrimp pond. This is especially true, given the recently recognized role of beneficial microbes in promoting shrimp nutrient intake and in conferring resistance against pathogens. Unfortunately, aquaculture-related microbiome studies are scarce in Southeast Asia countries despite the severe impact of early mortality syndrome outbreaks on shrimp production in the region. In this study, we employed the 16S rRNA amplicon (V3-V4 region) sequencing and amplicon sequence variants (ASV) method to investigate the microbial diversity of shrimp guts and pond water samples collected from aquaculture farms located in Malaysia and Vietnam. Substantial differences in the pond microbiota were observed between countries with the presence and absence of several taxa extending to the family level. Microbial diversity of the shrimp gut was found to be generally lower than that of the pond environments with a few ubiquitous genera representing a majority of the shrimp gut microbial diversity such as Vibrio and Photobacterium, indicating host-specific selection of microbial species. Given the high sequence conservation of the 16S rRNA gene, we assessed its veracity at distinguishing Vibrio species based on nucleotide alignment against type strain reference sequences and demonstrated the utility of ASV approach in uncovering a wider diversity of Vibrio species compared to the conventional OTU clustering approach.
Species of the benthic dinoflagellate Gambierdiscus produce polyether neurotoxins that caused ciguatera fish/shellfish poisoning in human. The toxins enter marine food webs by foraging of herbivores on the biotic substrates like macroalgae that host the toxic dinoflagellates. Interaction of Gambierdiscus and their macroalgal substrate hosts is believed to shape the tendency of substrate preferences and habitat specialization. This was supported by studies that manifested epiphytic preferences and behaviors in Gambierdiscus species toward different macroalgal hosts. To further examine the supposition, a laboratory-based experimental study was conducted to examine the growth, epiphytic behaviors and host preferences of three Gambierdiscus species towards four macroalgal hosts over a culture period of 40 days. The dinoflagellates Gambierdiscus balechii, G. caribaeus, and a new ribotype, herein designated as Gambierdiscus type 7 were initially identified based on the thecal morphology and molecular characterization. Our results showed that Gambierdiscus species tested in this study exhibited higher growth rates in the presence of macroalgal hosts. Growth responses and attachment behaviors, however, differed among different species and strains of Gambierdiscus over different macroalgal substrate hosts. Cells of Gambierdiscus mostly attached to substrate hosts at the beginning of the experiments but detached at the later time. Localized Gambierdiscus-host interactions, as demonstrated in this study, could help to better inform efforts of sampling and monitoring of this benthic toxic dinoflagellate.
This study was conducted to evaluate umami taste in protein hydrolysate produced from green mussel (Perna viridis) by hydrolysing with flavorzyme at pH 8, enzyme substrate ratio (E/S) 3% with or without the presence of 0.4% sodium tripolyphosphate (STPP) and 1.5% NaCI. Degree of hydrolysis (DH), yield, amino acid compositions, molecular weight distribution and sensory evaluation were determined. The highest DH (23.18%), darkest color and highest yield (8.34%) were recorded for hydrolysate produced in the presence of both STPP and NaCI. Electrophoresis analysis showed the presence of protein bands between 10 to 70 kDa where hydrolysate with addition of STPP and NaCI had bands with lower intensities. Amino acids which contribute to the umami taste such as glutamic acid, glycine and aspartic acid were higher in hydrolysate produced with STPP and NaCI addition. The hydrolysate has lesser fishy odor and flavor than those produced with only in the presence of flavorzyme and was also rated with highest score for all the five basic tastes including bitterness. However, the score for bitterness was still lower than the reference solutions. Therefore, green mussel hydrolysate produced in this study has a good potential as a food flavorant.
Metallic contamination in seafood, especially fish, has been of increasing concern to human health. Moreover, with increasing dependency on farmed fish for fish resources, the metallic contamination in them is still questionable. This study aimed to investigate the effects of cooking (steaming) on heavy metal concentration in farmed fish and to estimate its potential human health risk. Farmed sea basses (Lates calcarifer) from Setiu Lagoon were used to study the difference in metal uptake through human consumption of raw and cooked (steamed) fish samples. Selected heavy metals, namely copper (Cu), zinc (Zn), arsenic (As), cadmium (Cd), mercury (Hg) and lead (Pb), were measured using ICP-MS following Teflon bomb closed digestion of the fish samples. Cooking of the fish muscle by steaming was applied to investigate if cooking changes the concentration of heavy metals. Mercury and As were found accumulated more notably in the fish muscle, though only Hg was found to show significant (p > 0.05) increase when L. calcarifer is cooked. The amount of As in the fish muscle throughout its growth can be potentially harmful to humans, with the highest averaged concentration at 3.29 ± 0.65 mg/kg dw. above the standard set by the Malaysian Food Regulation (1985) of >1 mg/kg. All the other heavy metals were at relatively safe concentrations well below the standard set by both national and international guidelines. The PTWI per individual of L. calcarifer for As was at 0.84 mg/kg bw., which indicated that the amount of this fish safe for consumption without any adverse effect is 170 g/week. Therefore, long-term intake of these fish may pose a risk to human health due to the relatively higher Hg and As concentration found in these fishes.
Vibrio parahaemolyticus is a foodborne bacterial pathogen that may cause gastroenteritis in humans through the consumption of seafood contaminated with this microorganism. The emergence of antimicrobial and multidrug-resistant bacteria is another serious public health threat worldwide. In this study, the prevalence and antibiotic susceptibility test of V. parahaemolyticus in blood clams, shrimps, surf clams, and squids were determined. The overall prevalence of V. parahaemolyticus in seafood was 85.71% (120/140), consisting of 91.43% (32/35) in blood clam, 88.57% (31/35) in shrimps, 82.86% (29/35) in surf clams, and 80% (28/35) in squids. The majority of V. parahaemolyticus isolates from the seafood samples were found to be susceptible to most antibiotics except ampicillin, cefazolin, and penicillin. The MAR indices of V. parahaemolyticus isolates ranged from 0.04 to 0.71 and about 90.83% of isolates were found resistant to more than one antibiotic. The high prevalence of V. parahaemolyticus in seafood and multidrug-resistant isolates detected in this study could pose a potential risk to human health and hence appropriate control methods should be in place to minimize the potential contamination and prevent the emergence of antibiotic resistance.
Gelatin from salmon (Salmo salar) skin with high molecular weight protein chains (α-chains) was extracted using trypsin-aided process. Response surface methodology was used to optimise the extraction parameters. Yield, hydroxyproline content and protein electrophoretic profile via sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of gelatin were used as responses in the optimization study. The optimum conditions were determined as: trypsin concentration at 1.49 U/g; extraction temperature at 45 °C; and extraction time at 6 h 16 min. This response surface optimized model was significant and produced an experimental value (202.04 ± 8.64%) in good agreement with the predicted value (204.19%). Twofold higher yields of gelatin with high molecular weight protein chains were achieved in the optimized process with trypsin treatment when compared to the process without trypsin.
The present study was undertaken to determine whether Ligularia fischeri leaf extract (LF) is efficacious against collagen-induced arthritis (CIA) in mice. DBA/1J mice were immunized with bovine type II collagen and treated with LF (100 and 200 mg/kg) for 49 days. Mice were assessed regularly for signs of arthritis and the levels of rheumatoid factor, anti-type II collagen antibody, cytokines, AST, ALT, and creatinine in serum were also examined after the animals were killed. The arthritis score and paw edema were markedly suppressed in the groups treated with LF. Moreover, levels of rheumatoid factor, anti-type II collagen antibody, tumor necrosis factor-alpha, interleukin (IL)-1, and IL-6 in sera were reduced by LF administration. These data suggest that L. fischeri might be effective for the treatment of inflammatory arthritis like human rheumatoid arthritis.
Malaysian locally processed raw food products are widely used as main ingredients in local cooking. Previous studies showed that these food products have a positive correlation with the incidence of cancer. The cytotoxicity effect was evaluated using MTT assay (3-(4,5-dimetil-2-thiazolil)-2,5-diphenyl-2H-tetrazolium bromide) against Chang liver cells at 2000 microg/ml following 72 h incubation. Findings showed all methanol extracts caused a tremendous drop in the percentage of cell viability at 2000 microg/ml (shrimp paste - 41.69+/-3.36%, salted fish - 37.2+/-1.06%, dried shrimp - 40.32+/-1.8%, p<0.05). To detect DNA damage in a single cell, alkaline Comet Assay was used. None of the extracts caused DNA damage to the Chang liver cells at 62.5 microg/ml following 24 h incubation, as compared to the positive control, hydrogen peroxide (tail moment - 9.50+/-1.50; tail intensity - 30.50+/-2.50). Proximate analysis which was used for the evaluation of macronutrients in food showed that shrimp paste did not comply with the protein requirement (<25%) as in Food Act 1983. Salt was found in every sample with the highest percentage being detected in shrimp paste which exceeded 20%. Following heavy metal analysis (arsenic, cadmium, lead and mercury), arsenic was found in every sample with dried shrimps showing the highest value as compared to the other samples (6.16 mg/kg). In conclusion, several food extracts showed cytotoxic effect but did not cause DNA damage against Chang liver cells. Salt was found as the main additive and arsenic was present in every sample, which could be the probable cause of the toxicity effects observed.
The amino acid compositions of bovine, porcine and fish gelatin were determined by amino acid analysis using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate as derivatization reagent. Sixteen amino acids were identified with similar spectral chromatograms. Data pre-treatment via centering and transformation of data by normalization were performed to provide data that are more suitable for analysis and easier to be interpreted. Principal component analysis (PCA) transformed the original data matrix into a number of principal components (PCs). Three principal components (PCs) described 96.5% of the total variance, and 2 PCs (91%) explained the highest variances. The PCA model demonstrated the relationships among amino acids in the correlation loadings plot to the group of gelatins in the scores plot. Fish gelatin was correlated to threonine, serine and methionine on the positive side of PC1; bovine gelatin was correlated to the non-polar side chains amino acids that were proline, hydroxyproline, leucine, isoleucine and valine on the negative side of PC1 and porcine gelatin was correlated to the polar side chains amino acids that were aspartate, glutamic acid, lysine and tyrosine on the negative side of PC2. Verification on the database using 12 samples from commercial products gelatin-based had confirmed the grouping patterns and the variables correlations. Therefore, this quantitative method is very useful as a screening method to determine gelatin from various sources.
Scombroid fish poisoning is usually associated with consumption of fish containing high levels of histamine. However, reports indicate that some cases have responded to antihistamine therapy while ingested histamine levels in these cases were low. Potentiation of histamine toxicity by some biogenic amines, and release of endogenous histamine by other compounds such as cis-urocanic acid (UCA) are some hypotheses that have been put forth to explain this anomaly. Very little is known about the effects of storage conditions on the production of both UCA isomers and biogenic amines in tuna. Thus, the production of trans- and cis-UCA, histamine, putrescine, and cadaverine in tuna during 15 d of storage at 0, 3, and 10 °C and 2 d storage at ambient temperature were monitored. The initial trans- and cis-UCA contents in fresh tuna were 2.90 and 1.47 mg/kg, respectively, whereas the levels of putrescine and cadaverine were less than 2 mg/kg, and histamine was not detected. The highest levels of trans- and cis-UCA were obtained during 15 d storage at 3 °C (23.74 and 21.79 mg/kg, respectively) while the highest concentrations of histamine (2796 mg/kg), putrescine (220.32 mg/kg) and cadaverine (1045.20 mg/kg) were obtained during storage at room temperature, 10 and 10 °C, respectively. Histamine content increased considerably during storage at 10 °C whereas trans- and cis-UCA contents changed slightly. The initial trans-UCA content decreased during storage at ambient temperature. Thus, unlike histamine, concentrations of trans- and cis-UCA did not result in elevated levels during storage of tuna.
The concentration of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) present in the sediment and water of Peninsular Malaysia as well as in the cockle Anadara granosa was investigated. Samples were extracted and analysed with gas chromatography-mass spectrometry. The concentrations of total carcinogenic polycyclic aromatic hydrocarbons (t-PAHs) were measured between 0.80 +/- 0.04 to 162.96 +/- 14.74 ng/g wet weight (ww) in sediment, between 21.85 +/- 2.18 to 76.2 +/- 10.82 ng/L in water samples and between 3.34 +/- 0.77 to 46.85 +/- 5.50 ng/g ww in the cockle tissue. The risk assessment of probable human carcinogens in the Group B2 PAHs was calculated and assessed in accordance with the standards of the United States Environmental Protection Agency (US EPA). Case I in the toxicity assessment analysed the cancer risk to consumers of Malaysian blood cockle. Case II assessed the risk of cancer from exposure to PAHs from multiple pathways. The average cancer risk of case I and case II were found to be classifiable as unsafe according to the US EPA standard. The cancer risk due to c-PAHs acquired by the ingestion of blood cockle was (8.82 +/- 0.54) x 10-6 to (2.67 +/- 0.06) x 10(-2), higher than the US EPA risk management criterion. The non-cancer risks associated with multiple pathways in Kuala Gula, Kuala Juru and Kuala Perlis were higher than the U.S. EPA safe level, but the non-cancer risk for eating blood cockle was below the level of U.S. EPA concern.
Seafood is common item in the world diet; Asian countries have the highest rates of fish consumption in the world, which is higher than world average. Several studies have been conducted on the epidemiology and clinical characteristics of seafood allergy in different countries, and some of the fish and seafood allergens unique to those regions have been characterized. Review on published data showed that seafood allergy is very ubiquitous in some regions of the world. Fish and shellfish are the most common seafood that cause adverse allergic reactions among nations; the symptoms ranged from oral allergy syndromes to urticaria and anaphylaxis. The major identified allergens are parvalbumin in fish and tropomyosin in shellfish. Nevertheless, such studies are lacking from some regions with high fish and seafood consumption. Furthermore, the published data are mostly from small groups of populations, which large-scale epidemiological studies need to be performed.