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This study investigates the effect of antibiotics and sanitizers on biofilm forming Salmonella isolated from different seafood contact surfaces. Four Salmonella were isolated from 384 swab samples collected from various contact surfaces of fishing boats, fish landing centres and seafood processing plants. One out of four isolates was from the fishing boat (FB I -1) other three isolates were from the seafood processing plant (FPPII -4, FPPII- 5, FPPI-3). The ability of Salmonella to form biofilms on different contact surfaces (HDPE, stainless steel, wood, glass, tiles) was tested with the microbial load on different incubation days, and a higher count was observed on day five. The effect of sanitizer viz., sodium hypochlorite (20, 50, 100, 200 mg/l) and iodophor (2, 5, 10 mg/l) on the biofilm formed on different seafood contact surfaces were investigated. A reduction of 2-3 log was observed on surfaces of HDPE and stainless steel when they were treated with a minimum of 5 mg/l of iodophor or 20 mg/l of sodium hypochlorite after a contact time of 5 min. Antibiotic resistance of biofilm forming Salmonella was tested for different classes of antibiotics (penicillin, ß-lactams, quinolones, macrolides, aminoglycosides, phenol drugs, sulfonamides, cephalosporin). All four isolates showed intermediate resistance to ciprofloxacin, a quinolone drug. Only one isolate FB I -1 (fishing boat deck) expressed resistance to more drugs, viz., ßlactams (AMC, AMP, penicillin G), macrolides (AZM) and nitrofurantoin (NIT). These findings shall help the seafood processors to mitigate the formation of Salmonella biofilms on various seafood contact surfaces with different sanitizers and the antibiotic resistance of biofilm forming Salmonella shall give knowledge on human clinical treatments. With this study, we shall recommend the regulatory authorities control the contamination level of fish handling areas.

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The prevalence of biofilm forming Salmonella on different seafood contact surfaces was investigated. Out of 384 swab samples, 16.14 % and 1 % were confirmed biochemically and molecularly as Salmonella respectively. One out of four isolates was from the boat deck, and three were from the seafood processing plant. Salmonella was more prevalent in January, June, and September months. Different assays investigated the biofilm forming ability of isolates. Two out of four isolates have shown strong biofilms, and the others were moderate biofilm formers by microtitre plate assay. In the CRA assay, three isolates showed 'rdar' morphotype, and one showed 'bdar' morphotype. All isolates were positive for gcpA gene (~1700 bp), a critical gene found in Salmonella biofilms. The microbial load of Salmonella biofilms on different contact surfaces were determined, stainless steel and HDPE were found prone to biofilms. With this, a suitable mechanism shall be formulated to control the biofilms of Salmonella.

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Pesticide residues (PRs) in farmed shrimps are concerning food safety risks. Globally, India is a major exporter of pacific white leg shrimp (P. vannamei). This study was undertaken to analyze PRs in the water, sediments, shrimps, and feed at different growth stages to evaluate the ecotoxicological and human health risks. PRs in the seawater and sediments ranged from not detected (ND) to 0.027 µg/L and 0.006-12.39 µg/kg, and the concentrations were within the maximum residual limits (MRLs) and sediment quality guidelines prescribed by the World Health Organization and Canadian Environment Guidelines, respectively. PRs in shrimps at three growth stages viz. Postlarvae, juvenile, and adults, ranged from ND to 0.522 µg/kg, below the MRLs set by Codex Alimentarius Commission and European Commission. Most of the PRs in water, sediments, and shrimps did not vary significantly (p > 0.05) from days of culture (DOC-01) to DOC-90. The hazard quotient (HQ) and hazard ratio (HR) were found to be < 1, indicating that consumption of shrimps has no noncarcinogenic and carcinogenic risks. PRs in shrimp feed ranged from ND to 0.777 µg/kg and were found to be below the MRLs set by EC, which confirms that the feed fed is safe for aquaculture practices and does not biomagnify in animals. The risk quotient (RQ) and toxic unit (TU) ranged from insignificant level (ISL) to 0.509 and ISL to 0.022, indicating that PRs do not pose acute and chronic ecotoxicity to aquatic organisms. The study suggested no health risk due to PRs in shrimps cultured in India and exported to the USA, China, and Japan. However, regular monitoring of PRs is recommended to maintain a sustainable ecosystem.

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This study compared the heavy metal concentration in water, sediment, and shrimp at different growth stages of culture and subsequently evaluated the ecotoxicological and human health risk status. Total trace element concentration in the water, sediment and shrimp ranged from not detected (ND) (Hg) to 91.05 (Fe) µg/L, 0.01 (Hg) to 19, 246.33 (Fe) mg/kg, and ND (Hg) to 13.98 (Fe) mg/kg, respectively. Toxic metals such as, Cd, Hg, and Pb in shrimps ranged from ND to 2.11 mg/kg, ND to 0.158 mg/kg, ND to 0.088 mg/kg, and ND to 0.469 mg/kg, respectively. Toxic heavy metals at all the growth stages of shrimps (days of culture (DOC)-01 to DOC-90) were found below the maximum residual limit (MRL) of 0.5 mg/kg set by the European Commission (EC). Similarly, Cu, Zn, and As concentrations in shrimp were also far below the MRLs of 30 mg/kg, 100 mg/kg, and 76 mg/kg set by the World Health Organization and Food Safety and Standard Authority of India, respectively. The concentration of heavy metals increased from DOC-01 to DOC-90 and was positively correlated with the length and weight of the shrimps (p < 0.05). The risk assessment was estimated for both Indians and Americans and found no carcinogenic (lifetime cancer risk (LCR) < 10-4) and non-carcinogenic (THQ and TTHQ<1) health risks through consumption of shrimp cultured in this region. The hazard quotient (HQdermal < 1), hazard index (HI < 1), and LCR (<10-4) values of the heavy metals indicated that the dermal absorption might not be a concern for the local fishermen and marine fish/shrimp farmworkers. Water and sediment quality indices were applied to assess the surface water and sediment quality, and their results were found nil to low levels of heavy metal contamination at all the sampling sites. All heavy metals studied in sediments were < effect range low (ERL) and < threshold effect level (TEL), indicating no adverse biological effects on aquatic organisms. Therefore, regular monitoring of the shrimp aquaculture system throughout the crop will provide evidence of heavy metals bioaccumulation in shrimps. This research will provide baseline data to help farmers establish the optimal aquaculture practices and regulatory authorities to formulate legislation and strategies to reduce heavy metal biomagnification in shrimps from farm to fork.

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Food safety is crucial in today's competitive trading market, as it directly affects human health and promotes seafood exports. The effects of thermal processing (boiling, frying, grilling, and microwave cooking) on pesticide residues (PR) in P. vannamei were assessed. The PR in raw and processed shrimp ranged from 0.007 to 0.703 µg/kg for uncooked/raw, not detected (ND) to 0.917 µg/kg for boiled, ND to 0.506 µg/kg for fried, ND to 0.573 µg/kg for grilled and ND to 0.514 µg/kg for microwave cooked shrimps. The Endrin, endosulfan sulfate, and heptachlor were predominant PR found in the raw and processed shrimp. The PR content in raw and cooked shrimps were below the maximum residue limits (MRL) set by the Codex Alimentarius Commission (2021) and the European Commission (86/363/1986 and 57/2007). The estimated daily intake (EDI) of PR from raw and processed shrimps were below the ADI prescribed by CAC. The hazard quotient (HQ) and hazard ratio (HR) values were <1, indicating no non-carcinogenic or carcinogenic health implications through shrimp consumption. The estimated maximum allowable shrimp consumption rate (CRlim) suggests an adult can eat >100 shrimp meals/month, which is over the USEPA's (2000)recommendation of >16 meals/month without health issues. The Effect of thermal processing was detected in the following order: boiling < grilling < frying < microwave cooking. The processing factor (PF < 0.7), paired t-test (t < 0.05), Tukey post hoc (p < 0.05) test, Bray-Curtis similarity index, and matrix plot exhibited that all the four thermal processing methods have a considerable impact on pesticides in the processed shrimps. But frying (59.4%) and microwave cooking (60.3%) reduced PR far beyond boiling (48.8%) and grilling (51.3%). Hence, we recommend frying and microwave processing are better methods for minimizing PR in seafood than boiling or grilling.

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The concentrations of eight trace elements (chromium, cobalt, copper, zinc, arsenic, cadmium, mercury and lead) in14 commercially important fish and shellfish collected from Thoothukudi along the southeast coast of India was investigated using inductively coupled plasma mass spectrometry in order to assess the health risks associated with their consumption. The concentration of trace elements ranged from 0.001 to 39.5 µg/g. The estimated weekly intake of cadmium in seven fish and shellfish (0.0081-0.0996 mg/kg body weight) were above the provisional tolerable weekly intake set by the Joint FAO/WHO Expert Committee on Food Additives. The risk assessment analysis indicated that there was non- carcinogenic risk upon lifetime consumption of rock crab, C. natator (TTHQ >1) and carcinogenic risks upon lifetime consumption of S. jello, P. semisulcatus, P. sanguinolentus C. natator, Uroteuthis duvaceli, Sepia pharaonis and Cistopus indicus due to cadmium exposure indicating a potential health risk to the exposed consumers.

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Trace element pollution in the marine system is a global concern as the exposure of marine organisms to this pollution results in bioaccumulation and further transfer of the trace elements to humans through food chain. In the present study, the distribution of trace elements, namely chromium, cobalt, nickel, iron, copper, zinc, arsenic, cadmium, mercury, and lead, in gills, bone, liver, and muscle of eight commercially important fish collected along the southeast coast of India was analyzed using an inductively coupled plasma mass spectrometer. The liver was the main organ of accumulation for copper, zinc, arsenic, cadmium, and mercury; bone for chromium, cobalt, and lead; gills for copper; and muscle for arsenic and mercury. The concentration of toxic trace elements such as arsenic, cadmium, mercury, and lead in the edible portion of fish was lower than the recommended International Legislation limits, indicating that the fish of this region are safe for consumption.

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This study evaluates the distribution of pesticides and assesses the ecological and human health risks associated with pesticide residues concentration in the Thamirabarani River, the only perennial river in Tamil Nadu, India. Observed a variation in the pesticide concentration in the water (not detected (ND)-31.69 µg/L), sediments (ND-14.77 µg/kg), and fish (0.02-26.05 µg/kg). Endosulfan, aldrin, and endrin were the predominant organochlorine pesticides present in water, sediments, and fish. The average concentration of pesticides (except endosulfan) in water and sediments was found to be below the acceptable threshold as per the water and sediment quality guidelines, posing no ecological hazard to aquatic organisms. The calculated risk quotient and toxic unit (0.1 > TU/RQ ≤ 1) represent low-to-medium acute and chronic toxicity to the aquatic organisms inhabiting the river basin. The average concentration of pesticides in fish (Labeorohita) was also below the maximum residual limits set by the Codex Alimentarius Commission (CAC). However, the calculated daily intakes of endosulfan, aldrin, and endrin were above the CAC-acceptable daily intake guidelines. The human health risk assessment showed that children and adults exposed to pesticides in water and sediments through ingestion and dermal contact could have higher cancer risks (CR > 10-4) than inhalation. This study recommends implementing effective and routine pollution management schemes to avoid pesticide threats to aquatic and human health.

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Bioaccumulation of organochlorine pesticides (OCPs) in red, green, and brown seaweeds from the Gulf of Mannar was investigated using gas chromatography-mass spectrometry (GC-MS). The concentration of OCPs in all the seaweeds was in the order of endosulfan > endrin > HCH > DDT > aldrin > heptachlor > methoxychlor. The concentrations of HCH, endrin, endosulfan, and DDT in Sargassum wightii and Gelidiella acerosa, and HCH and endosulfan in Gracilaria verrucosa were above the limits prescribed by the European Commission. However, in Ulva lactuca, except for endosulfan, all other OCPs were below the permissible level. The bioaccumulation of OCPs in seaweeds was in the order of S. wightii > G. acerosa > Gracilaria verrucosa > U. lactuca. The results of comparative analysis of the levels of the investigated pesticides against the standard human health risk assessment revealed that pesticide exposure from seaweeds is hazardous to human health and the marine environment of the Gulf of Mannar.

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Industrialization and advancements in agriculture are increasingly the cause of environmental concerns and need to be addressed. This study was carried out at four sampling sites, viz. Manimuthar (site 1), Tirunelveli (site 2), Srivaikuntam (site 3), and Authoor (site 4), along the Thamirabarani river. The heavy metal concentration in fish and sediments were ranged from 0.001 to 9.505 mg kg-1 and 0.294 to 106.25 mg kg-1. The lifetime cancer risk (LCR) values for children and adults were found to be above the acceptable threshold value (ATV) and children were found to be more susceptible to health risks. For all sites except site 4, the LCR of toxic metals except Cd were within the ATV (10-6-10-4); the value of Cd was above the ATV, and poses a high cancer risk to the downstream inhabitants. This study advises routine heavy metal monitoring on aquatic organisms along these sites to implement regulatory standards.

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The prevalence of antibiotic-resistant Salmonella spp. in the source waters of shrimp farms in the Nagapattinam region of South India was investigated. Water and sediment samples (188 Nos.) collected from 23 natural creeks during December 2018 to April 2019 were examined for Salmonella spp. by conventional and PCR methods. The study indicated 28.7% of water and 25.5% of sediment isolates as Salmonella spp., while PCR test gave positive for 7.44% and 5.15% of the isolates, respectively. The isolates were resistant to sulfonamide (SF), but sensitive to tetracycline (TC), chloramphenicol (CAP), and furazolidone (FZ). PCR amplification of mitochondrial 16S rRNA region identified the highly resistant Salmonella serovar as S. Montevideo, which is an emerging food-borne pathogen. The incidence of antibiotic-resistant S. Montevideo reported for the first time in the natural creeks that supply water for shrimp farms emphasizes the need for regulatory steps to control its prevalence.

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Concentrations of trace elements (arsenic, cadmium, mercury, and lead) in tissues (muscle, gills, and digestive gland) of three commercially exploited crustaceans (Portunus sanguinolentus, Charybdis natator, and Penaeus semisulcatus) and three cephalopods (Doryteuthis sibogae, Sepia pharaonis, and Cistopus indicus) were examined. The animals were captured in the waters of Thoothukudi, and the tissues of six individuals of each species were analyzed using ICP-MS. The highest concentrations of arsenic (16.5 µg/g) and mercury (0.052 µg/g) were recorded in the digestive gland of C. natator, and cadmium (69.9 µg/g) and lead (0.351 µg/g) in the digestive gland of S. pharaonis. The edible portion of the crustaceans and cephalopods contained lower concentrations of trace elements, and these were below allowable limits set by the European Union. The edible parts of the investigated samples are safe for human consumption, but accidental contamination of the edible tissues with material from the digestive glands could lead to concerns relating to metal toxicity.

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Food authenticity is an issue of major concern for food authorities, as mislabeling represents one of the major commercial frauds. In this study, a novel PCR-RFLP protocol was developed as a tool to authenticate four shrimp products of commercial importance belonging to the family, Penaeidae, viz. Litopenaeus vannamei, Penaeus monodon, P. semisulcatus and Fenneropenaeus indicus. PCR amplification was performed targeting 16S rRNA/tRNAval region having an amplicon size of 530bp using the specific primers for shrimps, 16S-Cru4/16S-Cru3. Subsequent restriction analysis with a single restriction enzyme, Tsp5091, yielded distinct RFLP pattern for each species of shrimps having fragment sizes below 150bp. The unique RFLP patterns were also obtained in processed shrimp products without any degradation or alteration in the major fragments. The method was also validated with commercial shrimp products. Thus, the developed protocol can be performed within 8h using a single enzyme to authenticate four shrimp products of commercial significance.

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Squid protein hydrolysates (SPH) were prepared from the Indian squid Loligo duvauceli using papain. Response surface methodology (RSM) was used for optimization of hydrolysis conditions, including temperature, time, and the enzyme-substrate ratio using DPPH radical scavenging activity as a response. The amino acid composition of SPH was compared with raw squid muscle. In vitro antioxidant activities were evaluated based on reducing power, metal chelation, ABTS, hydroxyl radical, and superoxide anion radical scavenging assays. SPH exhibited good ABTS radical scavenging activities of 96.50±0.90%, superoxide anion radical scavenging activities of 96.4±0.89%, reducing powers of 0.71±0.02, moderate hydroxyl radical scavenging activities of 64.03±2.11%, and metal chelating activities of 52.04±1.02%. In vivo antioxidant activities determined using a sardine minced model system showed 42% reduction in formation of secondary oxidative products as thiobarbituric acid reactive substances (TBARS), almost equivalent to reduction by ascorbic acid of 41.42% at 400 ppm.

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A multiplex polymerase chain reaction (MPCR)-based assay was developed for the simultaneous detection of Vibrios using the genus-specific RNA polymerase subunit A (rpoA) gene and specific detection of toxin-producing Vibrio cholerae strains using two sets of primer based on cholera toxin subunit A (ctxA) and repeat in toxin subunit A (RtxA)-producing genes. The MPCR method developed is applicable to both the simultaneous and the two-step detection of genus Vibrio total and toxigenic V. cholerae species. This assay was specific as no amplification occurred with the other bacterial pathogens tested. The sensitivity of the assay was tested by artificially spiking the shrimp homogenate with the toxigenic strain of V. cholerae (NICED 16582) in different dilutions. The developed MPCR assay could detect three cells of V. cholerae in 12 h pre-enrichment in APW. The proposed method is rapid, sensitive, and specific for the detection of Vibrio genus as well as toxin-producing V. cholerae strains in environmental samples.

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