Gymnodinium catenatum is a cosmopolitan, bloom-forming dinoflagellate known to produce a suite of potent paralytic shellfish poisoning (PSP) toxins. Here, we revisit two major blooms of G. catenatum along the Fujianese Coast, China, in 2017 and 2018. The impact area of the 2017 bloom was larger than that of the 2018 event. Field sampling and remote satellite sensing revealed that alongshore transport driven by the southwest wind, as well as physical accumulation driven by the northeast wind, played important roles in the development and distribution of the two bloom events. The relationship between wind-induced hydrodynamic conditions and the unprecedented HAB events established in this study adds greatly to our understanding of algal bloom dynamics along the Fujianese coast. These results improve our ability to detect, track, and forecast G. catenatum blooms, thereby potentially minimizing the negative impacts of future HAB events.
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.
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.
The recurrent environmental and economic issues associated with the diminution of fossil fuels are the main impetus towards the conversion of agriculture, aquaculture and shellfish biomass and the wastes into alternative commodities in a sustainable approach. In this review, the recent progress on recovering and processing these biomass and waste feedstocks to produce a variety of value-added products via various valorisation technologies, including hydrolysis, extraction, pyrolysis, and chemical modifications are presented, analysed, and discussed. These technologies have gained widespread attention among researchers, industrialists and decision makers alike to provide markets with bio-based chemicals and materials at viable prices, leading to less emissions of CO2 and sustainable management of these resources. In order to echo the thriving research, development and innovation, bioresources and biomass from various origins were reviewed including agro-industrial, herbaceous, aquaculture, shellfish bioresources and microorganisms that possess a high content of starch, cellulose, lignin, lipid and chitin. Additionally, a variety of technologies and processes enabling the conversion of such highly available bioresources is thoroughly analysed, with a special focus on recent studies on designing, optimising and even innovating new processes to produce biochemicals and biomaterials. Despite all these efforts, there is still a need to determine the more cost-effective and efficient technologies to produce bio-based commodities.
Chitin, being the second most abundant biopolymer after cellulose, has been gaining popularity since its initial discovery by Braconot in 1811. However, fundamental knowledge and literature on chitin and its derivatives from insects are difficult to obtain. The most common and sought-after sources of chitin are shellfish (especially crustaceans) and other aquatic invertebrates. The amount of shellfish available is obviously restricted by the amount of food waste that is allowed; hence, it is a limited resource. Therefore, insects are the best choices since, out of 1.3 million species in the world, 900,000 are insects, making them the most abundant species in the world. In this review, a total of 82 samples from shellfish-crustaceans and mollusks (n = 46), insects (n = 23), and others (n = 13)-have been collected and studied for their chemical extraction of chitin and its derivatives. The aim of this paper is to review the extraction method of chitin and chitosan for a comparison of the optimal demineralization and deproteinization processes, with a consideration of insects as alternative sources of chitin. The methods employed in this review are based on comprehensive bibliographic research. Based on previous data, the chitin and chitosan contents of insects in past studies favorably compare and compete with those of commercial chitin and chitosan-for example, 45% in Bombyx eri, 36.6% in Periostracum cicadae (cicada sloughs), and 26.2% in Chyrysomya megacephala. Therefore, according to the data reported by previous researchers, demonstrating comparable yield values to those of crustacean chitin and the great interest in insects as alternative sources, efforts towards comprehensive knowledge in this field are relevant.
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.
Cockle (Anadara granosa) meat wash water precipitate was hydrolyzed using bromelain. Experiments were carried out to determine optimum conditions for temperature, enzyme concentration and hydrolysis time using response surface methodology (RSM) based on a central composite rotatable design (CCRD) to obtain the highest value of nitrogen content (NC) and degree of hydrolysis (DH). Results revealed that the optimum conditions for temperature, enzyme concentration and hydrolysis time were 33.7°C, 1.45% (E/S) and 28.42 hrs, respectively. At the optimum condition, hydrolysis of cockle meat wash water precipitate using bromelain resulted in a NC of 0.6% and DH of 48%. The NC and DH were significantly influenced by temperature, enzyme concentration and hydrolysis time. When the bromelain concentration, hydrolysis time and temperature were increased, the values of NC and DH also increased. The hydrolysate produced contained flavor compounds found in clam and oyster which were 3-methylbutanol and 1-pentanol. The compound 3-MCPD was not found in the hydrolysate.
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.
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.
The study was conducted to determine the mineral contents of 20 species of marine fish and 4 species of shellfish from the west coast of Peninsular Malaysia.Overall, the contents of micro minerals in all samples were below the permissible limits; except for oyster; with copper slightly higher than the limit set by FAO/WHO (1984), but below the limit set by Malaysian Food Regulations (1985); and zinc content higher than the limit set by Malaysian Food Regulations (1985), but below the limit set by FAO/WHO (1984). Meanwhile for macro minerals, most samples contained comparable sodium contents, significantly lower of potassium contents, higher calcium contents, and extremely higher of magnesium contents compared to the common ranges reported in the literatures. All samples were good sources of micro and macro minerals and could provide multi-health benefits if consumed in recommended amounts.
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.
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.
Vibrio parahaemolyticus is a main foodborne disease in seafood and generally seafood is
easily deteriorates in quality of color and flavor. In this study, clove (Syzygium aromaticum)
extract shows potent antibacterial activity against growth of antibiotics resistant Vibrio
parahaemolyticus on seafood samples (cockles and shrimps). Vibrio parahaemolyticus was
artificial contaminates on the samples with 106 CFU/ml. The samples were treated with different
concentration of cloves extract with 10 mg/ml which are 0.5%, 5% and 10% concentration
from methanol food grade extraction in 0 hr, 5 min, 10 min, 15 min, 30 min, 60 min and
120 min. Tab water and deionized water were selected as a negative control. As a result, the
amount of 10 % cloves managed to mitigates the number of V. parahaemolyticus on seafood
samples in 5 minutes and 15 min on both samples. Therefore, our results signify the fact that
cloves can be apply as natural sanitizer which could meet consumer demands for safe and
traditionally consumed either raw without any undesirable effect when applied in the seafood
system industries.
DNA damaging effect of the salted and fermented food products (salted fishes, dried shrimps and shrimp pastes) collected from three different locations in Malacca namely Pantai Puteri, Batang Tiga and Kelemak on the DNA of the Chang liver cells were evaluated via Alkaline Comet Assay. Treatment at 62.5 mg/ml following 24 hours of incubation was used based on the preliminary cytotoxicity data. Percentage of damage to the DNA was calculated using software for scoring based on the tail moment and tail intensity (severity of the DNA damage). Hydrogen peroxide was used as positive control at 0.1 mM following 30 minutes of incubation in 4 C. The results showed that the methanol extracts of shrimp pastes and salted fish from Pantai Puteri, exhibited a higher DNA damage (shrimp pastes - TM - 8.33 ± 2.19; TI - 31.67 ± 5.84, salted fishes - TM - 2.25 ± 0.86; TI - 9.25 ± 1.55) and were expressed as (shrimp pastes) 56.66 ± 8.74% of DNA damage and methanol salted fish extracts from the same location showed 13.00 ± 2.84% of the DNA damage on Chang liver cells compared to the other extracts. Values for methanol extract of shrimp pastes from Pantai Puteri were comparable to the positive control - Hydrogen peroxide (TM- 9.50 ± 1.50; TI - 30.50 ± 2.50). On the other hand, aqueous salted fishes extract from Pantai Puteri (TM - 1.33 ± 0.42; TI - 8.67 ± 2.42) and shrimp pastes extracts from Kelemak (methanol extract - TM -1.75 ± 0.15; TI -7.50 ± 0.50, aqueous extract - TM - 1.00 ± 0.00; TI - 5.00 ± 0.00) showed slightly high value for tail moment and tail intensity as compared to negative control (TM - 0.29 ± 0.05; TI - 2.50 ± 0.29). Values for methanol extracts of shrimp pastes from Pantai Puteri were comparable to the positive control (TM- 9.50 ± 1.50; TI - 30.50 ± 2.50). In conclusion, our results demonstrate genotoxic damage induced by few salted and fermented food extracts in Chang liver cell.
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.
This study was undertaken to assess the levels of trace metals (As, Cd, Cu, Pb, and Zn) in two common species of cockles (Anadara granosa and Anadara inaequivalvis) from two coastal areas in Thailand (Pattani Bay) and Malaysia (the Setiu Wetlands). A total of 350 cockles were collected in February and September 2014. Trace metals were determined by Inductively Coupled Plasma Mass Spectrometry. We observed that cockles in both areas had a higher accumulation of metals in September. Notably, the biota-sediment accumulation (BSAF) of Cd was highest in both areas. A strong positive correlation of Cd with the length of the cockles at Pattani Bay (r(2) = 0.597) and the Setiu Wetlands (r(2) = 0.675) was noted. It was suggested that As could be a limiting element (BSAF < 1) of cockles obtained from Pattani Bay. In comparison with the permissible limits set by the Thailand Ministry of Public Health and the Malaysia Food Regulations, mean values of As, Cd, Cu, Pb, and Zn were within acceptable limits, but the maximum values of Cd and Pb exceeded the limits for both areas. Regular monitoring of trace metals in cockles from both areas is suggested for more definitive contamination determination.
The accumulation of paralytic shellfish toxins (PSTs) in contaminated shellfish is a serious health risk making early detection important to improve shellfish safety and biotoxin management. Capillary electrophoresis (CE) has been proven as a high resolution separation technique compatible with miniaturization, making it an attractive choice in the development of portable instrumentation for early, on-site detection of PSTs. In this work, capillary zone electrophoresis (CZE) with capacitively coupled contactless conductivity detector (C(4)D) and UV detection were examined with counter-flow transient isotachophoresis (tITP) to improve the sensitivity and deal with the high conductivity sample matrix. The high sodium concentration in the sample was used as the leading ion while l-alanine was used as the terminating electrolyte (TE) and background electrolyte (BGE) in which the toxins were separated. Careful optimization of the injected sample volume and duration of the counter-flow resulted in limit of detections (LODs) ranging from 74.2 to 1020 ng/mL for tITP-CZE-C(4)D and 141 to 461 ng/mL for tITP-CZE-UV, an 8-97 fold reduction compared to conventional CZE. The LODs were adequate for the analysis of PSTs in shellfish samples close to the regulatory limit. Intra-day and inter-day repeatability values (percentage relative standard deviation, n=3) of tITP-CZE-C(4)D and tITP-CZE-UV methods for both migration time and peak height were in the range of 0.82-11% and 0.76-10%, respectively. The developed method was applied to the analysis of a contaminated mussel sample and validated against an Association of Official Analytical Chemists (AOAC)-approved method for PSTs analysis by high performance liquid chromatography (HPLC) with fluorescence detection (FLD) after pre-column oxidation of the sample. The method presented has potential for incorporation in to field-deployable devices for the early detection of PSTs on-site.
Paralytic shellfish toxins (PSTs) are produced by marine and freshwater microalgae and accumulate in shellfish including mussels, oysters, and scallops, causing possible fatalities when inadvertently consumed. Monitoring of PST content of shellfish is therefore important for food safety, with currently approved methods based on HPLC, using pre- or postcolumn oxidation for fluorescence detection (HPLC-FLD). CE is an attractive alternative for screening and detection of PSTs as it is compatible with miniaturization and could be implemented in portable instrumentation for on-site monitoring. In this study, CE methods were developed for C(4) D, FLD, UV absorption detection, and MS-making this first report of C(4) D and FLD for PSTs detection. Because most oxidized toxins are neutral, MEKC was used in combination with FLD. The developed CZE-UV and CZE-C(4) D methods provide better resolution, selectivity, and separation efficiency compared to CZE-MS and MEKC-FLD. The sensitivity of the CZE-C(4) D and MEKC-FLD methods was superior to UV and MS, with LOD values ranging from 140 to 715 ng/mL for CZE-C(4) D and 60.9 to 104 ng/mL for MEKC-FLD. With the regulatory limit for shellfish samples of 800 ng/mL, the CZE-C(4) D and MEKC-FLD methods were evaluated for the screening and detection of PSTs in shellfish samples. While the CZE-C(4) D method suffered from significant interferences from the shellfish matrix, MEKC-FLD was successfully used for PST screening of a periodate-oxidized mussel sample, with results confirmed by HPLC-FLD. This confirms the potential of MEKC-FLD for screening of PSTs in shellfish samples.
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.
Detection of diatom frustules in bone marrow (diatom test) is used for diagnosing ante-mortem drowning where the usual signs of drowning are not present in dead bodies recovered from water. However, controversies over the reliability of diatom test results are continuing. There have been indications on the possibilities of diatoms entering into systemic circulation from atmospheric air, food and drink. While diatoms have been demonstrated in the gut content of edible marine forms such as shrimps and clams, the present study, for the first time, provides empirical evidence on the prevalence as well as abundance of diatom frustules in the samples of cooked non-vegetarian foodstuffs that impend human consumption in Kelantan, Malaysia. It is found that 50 g each of cleaned and cooked prawns and of clams impending human consumption contain about 8360 and 29,054 diatom frustules, respectively. A person accustomed to prawn and clam food would be ingesting an estimated 2 million diatoms in a single year. Considering the suggestion that detection of five diatom frustules in 10 g of bone marrow would suffice for concluding drowning as mode of death, and the fact that there is yet no proof that diatom frustules do not enter into the human systemic circulation through the digestive tract, the estimated number of diatom frustules routinely ingested acquires significance since entry of a few of such ingested frustules into the systemic circulation can lead to false positive test results. The findings of this research raise two important issues: first, population based routine food related diatom ingestion requires to be estimated, and, second, studies have to be initiated to categorically prove or disprove the possibility of entry of diatom frustules into the systemic circulation via the digestive tract.