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 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.
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.
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.
Antibiotic susceptibility, plasmid profiles and random amplification of polymorphic DNA (RAPD) were used to study strains of Vibrio vulnificus isolated from cockles (Anadara granosa). Thirty-six isolates were analyzed. The prevalent biotypes were 1 (72.2% of the isolates) and 2 (27.8%). Among these, 21 strains of biotype 1 and two strains of biotype 2 contained plasmid DNA bands ranging in size from 1.4 to 9.7 MDa. Thirty-one (83.3%) were found to be resistant to one or more of the antimicrobial agents tested, however no specific correlation between antimicrobial resistance patterns and a single biotype was found. In addition, no particular plasmid profile was predictive of a particular pattern of antibiotic susceptibility. Two primers produced polymorphisms in all strains tested, producing bands ranging from 0.25 to 2.7 kb, indicating a high variability among both biotype 1 and biotype 2 of the V. vulnificus strains investigated. RAPD identity across biotypes was also observed among Vibrio vulnificus strains.
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 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.
Green mussel (Perna viridis) was hydrolysed with alcalase under two different conditions consisting of pH7, E/S5% or pH 9, E/S 3% at 60°C for two hours. Hydrolysis at pH 9, E/S3% resulted in a higher degree of hydrolysis (DH) than pH7, E/S5% with degree of hydrolysis of 37.00% and 28.33%, respectively. The green mussel hydrolysates were characterized by molecular weight of
Vibrio parahaemolyticus is recognized as a frequent causal agent of human gastroenteritis due to the consumption of raw, undercooked or mishandled seafood in many Asian countries. The number of V. parahaemolyticus cases reported is on the rise, and this becomes a concern to the Asian countries as seafood is favoured by Asians. This study aimed to detect and quantify V. parahaemolyticus in raw oysters and to determine the risk associated with the consumption of raw oysters. A total of 30 oyster samples were collected and analysed in this study. MPN-PCR and MPN-Plating methods were employed and carried out concurrently to determine the prevalence of V. parahaemolyticus in raw oysters. The results showed that the prevalence of total V. parahaemolyticus in oysters was 50.00% (15/30) where the MPN/g range was < 3 – > 11000 MPN/g for MPN-PCR method, and 40.00% (12/30) where the MPN/g range was < 3 – 4300 MPN/g for MPN-Plating method. MPN-PCR method was able to estimate the level of virulence (tdh+ and trh+) V. parahaemolyticus in the raw oysters where 10.00% (3/30) of samples were identified to be in a range of 3 – 30 MPN/g. A microbial risk assessment was conducted based on the enumeration data obtained from MPN-PCR method using @risk. The probability of illness annually was 1.76 X 10-6 with a prediction of 31 cases to occur with respect to the exposed Malaysian population, while the rate per 100,000 people was estimated to be at 0.104. In addition, the antibiogram of V. parahaemolyticus was determined using Kirby Bauer Disk Diffusion Test and the results indicated that the isolates were highly resistant towards Bacitracin (100.00%), Vancomycin (100.00%) and were least resistant to Chloramphenicol (8.70%). The MAR index of the isolates ranged from 0.17 to 0.50. In accordance with the results from this study, the consumption of raw oysters is a risk factor for V. parahaemolyticus infection and proactive actions should be taken to reduce the risk of the pathogen in order to improve public health.
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.
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.
The worldwide depletion of major fish stocks through intensive industrial fishing is thought to have profoundly altered the trophic structure of marine ecosystems. Here we assess changes in the trophic structure of the English Channel marine ecosystem using a 90-year time-series (1920-2010) of commercial fishery landings. Our analysis was based on estimates of the mean trophic level (mTL) of annual landings and the Fishing-in-Balance index (FiB). Food webs of the Channel ecosystem have been altered, as shown by a significant decline in the mTL of fishery landings whilst increases in the FiB index suggest increased fishing effort and fishery expansion. Large, high trophic level species (e.g. spurdog, cod, ling) have been increasingly replaced by smaller, low trophic level fish (e.g. small spotted catsharks) and invertebrates (e.g. scallops, crabs and lobster). Declining trophic levels in fisheries catches have occurred worldwide, with fish catches progressively being replaced by invertebrates. We argue that a network of fisheries closures would help rebalance the trophic status of the Channel and allow regeneration of marine ecosystems.
Allergy diagnosis needs to be improved in polysensitized patients due to the existence of possible confounding factors in this type of patients. Component resolved diagnosis (CRD) is a new concept in the investigation of polysensitized patients. The aim of this study was to evaluate if the utilization of ImmunoCAP ISAC improve the diagnosis of the polysensitized allergic rhinitis patients. Skin prick test (SPT) to 58 crude allergen extracts and CRD (ImmunoCAP ISAC) were carried out for 5 polysensitized allergic rhinitis patients. Two patients had a shellfish allergy and avoidance of shellfish was the only way to prevent an allergic reaction. In contrast, although the remaining three patients had low risk for shellfish allergy, but they were the best candidates for immunotherapy using mite extracts. CRD and particularly ImmunoCAP ISAC have proven to be a valuable diagnostic tool in polysensitized patients. ImmunoCAP ISAC helps refine the individual patient's sensitization profile and predict the potential risk of allergic reactions and improve the selection of patients for immunotherapy.
Snail is one of the worst causes of food allergy. Thus, the aim of this study was to identify the major and minor allergens of the local marine snail (Cerithidea obtusa) and subsequently to investigate the impacts of heat treatment on the IgE-binding activity of snail allergens. Proteins from raw and heat-treated snails (boiled, roasted and fried) were extracted and then resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblotting of all extracts were then performed using sera from patients with snail allergy. The results showed that the raw extract contains numerous protein bands between 12 to>250 kDa. Some thermostable proteins, predominantly the 33 and 42 kDa bands, remained detected in all cooked extracts with decreasing intensities from boiled to roasted to fried extracts, while the majority of thermolabile bands denatured after heating. Boiled snail had more protein bands compared to roasted and fried snails. Immunoblotting of raw extract demostrated 19 IgE-binding bands ranging from 15 to 240 kDa. The thermostable bands of 33 and 42 kDa and a thermolabile of 30 kDa band were identified as the major allergens of this snail. The cooked extracts yielded less allergenic bands. The boiled extract yielded approximately 14 IgE-binding bands with some smeared bands at high molecular weight regions. The roasted extract had lesser IgE-binding bands and the majority appeared as smears, while the IgE-reactivity in the fried extract was less visible and appeared as weak smears. This study indicated that both raw and cooked snails played a crucial role in snail allergenicity, as this species of snail contains both thermostable and thermolabile major allergens. The degree of snail allergenicity was revealed in the order: raw> boiled > roasted> fried. Thus, the results would facilitate in the development of effective diagnosis and management strategies of snail allergy in this country.
Allergy caused by food is usually type 1 allergy of four types of allergic reactions. One of the most widespread allergic is those that are caused by crustacean shellfish. Crustaceans are classified among arthropods which include crab, crayfish, lobster, prawn and shrimp. Shrimp which are broadly consumed as nutritional food is one of the most important food that contribute to allergy. Thus, reducing the allergenicity of shrimp allergen will be helpful to individuals who are sensitive to shrimp and for this reason the characteristics of each allergen need to be studied. Those sensitized individuals can develop urticaria, angiodema, laryngospasm, asthma and life threatening anaphylaxis. To date, four main allergens contribute to allergic reactions. They are tropomyosin (TM), a highly conserved and heat stable myofibrillar protein of 35-38 kDa followed by arginine kinase (AK) which is also known as Pen m 2 or Lit v 2 with 40 kDa. Two other contributing allergens are sarcoplasmic calcium-binding protein (SCP) also known as Lit v 4 with 22 kDa and myosin light chain (MLC) which is also termed as Lit v 3 with 20 kDa. This mini-review will provide a better understanding of each allergen derived from shrimp which subsequently will help to reduce the allergenicity.
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.