Behavior of diclofenac from contaminated fish after cooking and in vitro digestion.

BACKGROUND: Seafood consumers are widely exposed to diclofenac due to the high contamination levels often present in aquatic organisms. It is a potential risk to public health due its endocrine disruptor properties. Limited information is available about diclofenac behavior after food digestion to enable a more realistic scenario of consumer exposure. This study aimed to evaluate cooking effects on diclofenac levels, and determine diclofenac bioaccessibility by an in vitro digestion assay, using commercial fish species (seabass and white mullet) as models. The production of the main metabolite 4'-hydroxydiclofenac was also investigated. Fish hamburgers were spiked at two levels (150 and 1000 ng g-1) and submitted to three culinary treatments (roasting, steaming and grilling). RESULTS: The loss of water seems to increase the diclofenac levels after cooking, except in seabass with higher levels. The high bioaccessibility of diclofenac (59.1-98.3%) observed in both fish species indicates that consumers' intestines are more susceptible to absorption, which can be worrisome depending on the level of contamination. Contamination levels did not affect the diclofenac bioaccessibility in both species. Seabass, the fattest species, exhibited a higher bioaccessibility of diclofenac compared to white mullet. Overall, cooking decreased diclofenac bioaccessibility by up to 40% in seabass and 25% in white mullet. The main metabolite 4'-hydroxydiclofenac was not detected after cooking or digestion. CONCLUSION: Thus, consumption of cooked fish, preferentially grilled seabass and steamed or baked white mullet are more advisable. This study highlights the importance to consider bioaccessibility and cooking in hazard characterization studies. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Assessing metal(loid)s concentrations and biomarkers in tilapia (Oreochromis niloticus) and largemouth bass (Micropterus salmoides) of three ecosystems of the Yaqui River Basin, Mexico.

Aquatic ecosystems have been suffering deleterious effects due to the development of different economic activities. Metal(loid)s are one of the most persistent chemicals in environmental reservoirs, and may produce adverse effects on different organisms. Since fishes have been largely used in studies of metal(loid)s exposure, tilapia and largemouth bass were collected in three ecosystems from the Yaqui River Basin to measure the concentrations of metal(loid)s (chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), zinc (Zn) arsenic (As), mercury (Hg), and selenium (Se)) and some biomarkers (somatic indices, metallothionein expression and histopathological analysis) in tissues of both species. Metal(loid) concentrations varied seasonally among ecosystems in tissues of both species. The elements varied seasonally and spatially in tissues of both species, with a general distribution of liver > gills > gonads. Also, biomarkers showed variations indicative that the fish species were exposed to different environmental stressor conditions. The highest values of some biomarkers were in largemouth bass, possibly due to differences in their biological characteristics, mainly feeding habits. The multivariate analysis showed positive associations between metal(loid)s and biomarkers, which are usually associated to the use of these elements in metabolic and/or regulatory physiological processes. Both fish species presented histological damage at different levels, from SI types (changes that are reversible for organ structure) to SII types (changes that are more severe but may be repairable). Taken together, the results from this study suggest that the Yaqui River Basin is moderately impacted by metals and metalloids.

Molecular cloning, inducible expression and function analysis of Epinephelus coioides Sec6 response to SGIV infection.

Exocyst complex component 3 Sec6 of mammals, one of the components of the exocyst complex, participates in numerous cellular functions, such as promoting cell migration and inhibiting apoptosis. In this study, the Sec6 was obtained from Epinephelus coioides, an economically important cultured fish. The full length of E. coioides Sec6 was 2655 bp including a 245 bp 5' UTR, a 154 bp 3' UTR, and a 2256 bp open reading frame (ORF) encoding 751 amino acids, with a molecular mass of 86.76 kDa and a theoretical pI of 5.57. Sec6 mRNA was detected in all the tissues examined, but the expression level is different in these tissues. Using fluorescence microscopy, Sec6 were distributed in both the nucleus and the cytoplasm. After SGIV infection, the expression of E. coioides Sec6 was significantly up-regulated in both trunk kidney and spleen response to Singapore grouper iridovirus (SGIV), an important pathogens of E. coioides. Sec6 could increase the SGIV-induced cytopathic effects (CPE), the expression of the SGIV genes VP19, LITAF, MCP, ICP18 and MCP, and the viral titers. Besides, E. coioides Sec6 significantly downregulated the promoter of NF-κB and AP-1, and inhibited the SGIV-induced apoptosis. The results demonstrated that E. coioides Sec6 might play important roles in SGIV infection.

Tolerance of juvenile Peruvian rock seabass (Paralabrax humeralis Valenciennes, 1828) and Peruvian grunt (Anisotremus scapularis Tschudi, 1846) to low-oxygen conditions.

Hypoxia is currently one of the greatest threats to coastal ecosystems worldwide, generating massive mortality of marine organisms, loss of benthic ecosystems and a decrease in fishery production. We evaluated and compared the tolerance to hypoxia of two species from different habitats of the Peruvian coast, the Peruvian rock seabass Paralabrax humeralis and the Peruvian grunt Anisotremus scapularis. The effect of hypoxia was measured as a function of the exposure time (progressive and chronic) on the behavioural and physiological responses of the two species, as well as on the enzymatic activity associated with the oxidative stress response of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and alkaline phosphatase (AKP). The ventilatory frequency was measured at two different temperatures (16 and 22°C) under progressive hypoxia conditions to determine the ventilatory critical point (Vcp). A. scapularis showed a higher Vcp than P. humeralis, which was positively affected by temperature. The median lethal time of A. scapularis was 36 min at 60% of oxygen saturation, while P. humeralis showed no mortality after 31 days of exposure at 5% oxygen saturation. Different enzymatic activity (P < 0.05) between species under hypoxia was recorded, in SOD (gill and muscle) and AKP (blood). A general tendency, under hypoxia, to slightly increase LDH activity (except for blood in A. scapularis, P < 0.05) and SOD activity (mainly in muscle of A. scapularis, P < 0.05), and decrease AKP activity (mainly in liver of P. humeralis, P < 0.05) was observed. The response of P. humeralis to hypoxia goes through a reduction in activity and metabolism, so this species can be considered hypoxia-tolerant, allowing it to face hypoxia events during prolonged periods. On the other hand, A. scapularis response to hypoxia prioritizes avoidance mechanisms and, together with other adaptations, makes it especially vulnerable to hypoxia and able to be considered hypoxia-intolerant.

Citral as food additive for common snook - zootechnical parameters and digestive enzymes / Citral como aditivo na ração para robalo flecha - parâmetros zootécnicos e enzimas digestivas

Essential oils of plants whose main compound is citral showed beneficial effects when added to fish feed. The objective of the present study was to evaluate the dietary effect of the addition of citral on zootechnical parameters and digestive enzyme activities of Centropomus undecimalis. Juveniles were fed for 45 days with diets containing different amounts of citral (0.0 - control, 0.5, 1.0, and 2.0 mL per kg of diet). The water quality parameters were kept stable during the experiment and no mortality was observed. At the end of the experimental period, the treatment 0.5 mL citral per kg of diet had the lowest weight gain and specific growth rate, and the highest feed conversion, while the same parameters did not differ between the other treatments. Pepsin activity was higher in the stomach of fish fed with 0.5 mL citral per kg of diet and amylase activity was higher in the intestine of fish fed with 0.5 and 2.0 mL citral per kg of diet compared to the control group. Intestinal lipase activity was higher in all groups that were fed with citral compared to the control group. Chymotrypsin and trypsin activities showed no difference between groups. Consequently, dietary addition of citral at any of the levels tested is not recommended for common snook.(AU)

Óleos essenciais de plantas, cujo composto majoritário é o citral, mostraram efeitos benéficos quando adicionados à ração para peixes. O objetivo do presente estudo foi avaliar o efeito da adição de citral sobre os parâmetros zootécnicos e atividades das enzimas digestivas de Centropomus undecimalis. Os juvenis foram alimentados por 45 dias com dietas contendo diferentes quantidades de citral (0,0 - controle, 0,5, 1,0 e 2,0 mL por kg de ração). Os parâmetros de qualidade da água foram mantidos estáveis ​​durante o experimento e nenhuma mortalidade foi observada. Ao final do período experimental, o tratamento 0,5 mL citral por kg de ração teve o menor ganho de peso e taxa de crescimento específico e a maior conversão alimentar, enquanto os mesmos parâmetros não diferiram entre os demais tratamentos. A atividade da pepsina foi maior no estômago de peixes alimentados com 0,5 mL de citral por kg de ração e a atividade de amilase foi maior no intestino de peixes alimentados com 0,5 e 2,0 mL citral por kg de ração comparado ao grupo controle. A atividade da lipase intestinal foi maior em todos os grupos que foram alimentados com citral, em comparação ao grupo controle. As atividades de quimotripsina e tripsina não mostraram diferença entre os grupos. Consequentemente, a adição de citral na ração em qualquer um dos níveis testados não é recomendada para robalo flecha.(AU)

Effects of Probiotic Bacteria Bacillus on Growth Performance, Digestive Enzyme Activity, and Hematological Parameters of Asian Sea Bass, Lates calcarifer (Bloch).

This study was conducted to evaluate different doses of two species of Bacillus (Bacillus licheniformis and Bacillus subtilis), on growth parameters, chemical composition of fish, activity of liver, and digestive enzymes of Asian sea bass. During 8 weeks, juvenile Asian sea bass received diets supplemented with 1 × 103, 1 × 106, and 1 × 109 CFU g-1 probiotic in addition to a control diet without added microorganisms. At the end of the trial, growth indices (total weight, total length, specific growth rate, total weight gain, food conversion ratio, and condition factor), body composition (crude protein, crude lipid, ash, and dry matter), digestive enzymes (protease, lipase, and amylase), liver enzymes [aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP)], immunologic indicators (lysozyme), and hematological parameters [hematocrit (Hct), hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs)] were assessed. Asian sea bass receiving diets supplemented with probiotic Bacillus (Bacillus licheniformis and Bacillus subtilis) showed significantly better growth than those fed the basal diet (control). Regarding body composition, total protein levels and dry matter were higher and lipid levels were lower in fish fed the diet containing 1 × 106 CFU g-1 probiotic compared with the control group (P < 0.05). Digestive enzymes (protease, lipase, and amylase) and hematological parameters (RBC, WBC, and Hb) were all highest in fish fed diet supplemented with 1 × 106 CFU g-1 probiotic Bacillus. Also, liver enzymes (AST, ALT, ALP) were lower in fish fed diet supplemented with 1 × 106 CFU g-1 probiotic Bacillus. Being that supplementation of 1 × 106 CFU g-1 of Bacillus in the diet is the dose which delivers the best results.

Digestibilidade, taxa de retenção proteica e excreção de amônia em juvenis de robalo-peva (Centropomus parallelus) alimentados com rações com diferentes níveis de proteína / Digestibility, protein retention rate and ammonia excretion in juvenile fat snook (Centropomus parallelus) fed with different protein levels

The aim of this study was to determine the level of dietary crude protein (CP) (480, 400, 440, 520 and 560g kg-1) in rations for juvenile II fat snook (Centropomus parallelus) that best promotes protein absorption and lower excretion of nitrogenous products. Three experiments were conducted: the first assessed the protein retention rate; the second evaluated ammonia excretion; and the third determined the best digestibility coefficient. The estimated protein retention rate showed a positive quadratic response to dietary CP with maximum retention at the 510.20g kg-1 protein level, and increasing levels of protein resulted in a greater total ammonia excretion rate in water. The mean apparent protein digestibility coefficient values increased with increasing levels of dietary protein up to 520g kg-1, where they stabilized. As determined by regression, the level of dietary CP that promotes the best digestibility was 495.62g kg-1.(AU)

O objetivo da pesquisa foi determinar o nível de proteína bruta na dieta (400, 440, 480, 520 e 560g kg-1), nas rações de juvenis II de robalo-peva (Centropomus parallelus), que promove melhor absorção de proteína e menor excreção de produtos nitrogenados. Foram realizados três experimentos. O primeiro avaliou a taxa de retenção de proteína, o segundo a excreção de amônia, e o terceiro determinou o melhor coeficiente de digestibilidade. Os dados foram analisados por meio de regressão exponencial. A taxa de retenção proteica, calculada pela equação, apresentou efeito quadrático positivo e o máximo de retenção com 510,20g kg-1 de proteína na dieta. O aumento dos níveis de proteína resultou em maiores taxas de excreção de amônia total na água. Os valores médios dos coeficientes de digestibilidade aparente da proteína foram aumentando com o aumento da proteína dietética até o nível de 520g kg-1 onde se estabilizou. O nível de proteína bruta na dieta, determinado pela equação de regressão, que promove a melhor digestibilidade é de 495,62g kg-1.(AU)

Digestibilidade, taxa de retenção proteica e excreção de amônia em juvenis de robalo-peva (Centropomus parallelus) alimentados com rações com diferentes níveis de proteína / Digestibility, protein retention rate and ammonia excretion in juvenile fat snook (Centropomus parallelus) fed with different protein levels

The aim of this study was to determine the level of dietary crude protein (CP) (480, 400, 440, 520 and 560g kg-1) in rations for juvenile II fat snook (Centropomus parallelus) that best promotes protein absorption and lower excretion of nitrogenous products. Three experiments were conducted: the first assessed the protein retention rate; the second evaluated ammonia excretion; and the third determined the best digestibility coefficient. The estimated protein retention rate showed a positive quadratic response to dietary CP with maximum retention at the 510.20g kg-1 protein level, and increasing levels of protein resulted in a greater total ammonia excretion rate in water. The mean apparent protein digestibility coefficient values increased with increasing levels of dietary protein up to 520g kg-1, where they stabilized. As determined by regression, the level of dietary CP that promotes the best digestibility was 495.62g kg-1.

O objetivo da pesquisa foi determinar o nível de proteína bruta na dieta (400, 440, 480, 520 e 560g kg-1), nas rações de juvenis II de robalo-peva (Centropomus parallelus), que promove melhor absorção de proteína e menor excreção de produtos nitrogenados. Foram realizados três experimentos. O primeiro avaliou a taxa de retenção de proteína, o segundo a excreção de amônia, e o terceiro determinou o melhor coeficiente de digestibilidade. Os dados foram analisados por meio de regressão exponencial. A taxa de retenção proteica, calculada pela equação, apresentou efeito quadrático positivo e o máximo de retenção com 510,20g kg-1 de proteína na dieta. O aumento dos níveis de proteína resultou em maiores taxas de excreção de amônia total na água. Os valores médios dos coeficientes de digestibilidade aparente da proteína foram aumentando com o aumento da proteína dietética até o nível de 520g kg-1 onde se estabilizou. O nível de proteína bruta na dieta, determinado pela equação de regressão, que promove a melhor digestibilidade é de 495,62g kg-1.

Characterization of quality of sediments from Paranaguá Bay (Brazil) by combined in vitro bioassays and chemical analyses.

The present study characterizes the quality of sediments from the Paranaguá Estuarine Complex (South Brazil). Polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and organochlorine pesticides (OCPs) were determined in sediment samples together with a series of different in vitro bioassays. The fish hepatoma cell line (PLHC-1) was used to determine the presence of cytotoxic compounds and CYP1A- and oxidative stress-inducing agents in sediment extracts. Ovarian microsomal fractions from sea bass (Dicentrarchus labrax) were used to detect the presence of endocrine disrupters that interfered with the synthesis of estrogens (ovarian CYP19). Despite the relatively low levels of pollutants and no evidence of negative effects based on guideline levels, sediments collected close to harbors were enriched with CYP1A-inducing agents and they showed higher cytotoxicity. In contrast, sediments from internal areas inhibited CYP19 activity, which suggests the presence of endocrine disrupters at these sites. Overall, the selected bioassays and the chemistry data led to the identification of potentially impacted areas along the Paranaguá Estuarine Complex that would require further action to improve their environmental quality. Environ Toxicol Chem 2017;36:1811-1819. © 2016 SETAC.

Seafood substitutions obscure patterns of mercury contamination in Patagonian toothfish (Dissostichus eleginoides) or "Chilean sea bass".

Seafood mislabeling distorts the true abundance of fish in the sea, defrauds consumers, and can also cause unwanted exposure to harmful pollutants. By combining genetic data with analyses of total mercury content, we have investigated how species substitutions and fishery-stock substitutions obscure mercury contamination in Patagonian toothfish (Dissostichus eleginoides), also known as "Chilean sea bass". Patagonian toothfish show wide variation in mercury concentrations such that consumers may be exposed to either acceptable or unacceptable levels of mercury depending on the geographic origins of the fish and the allowable limits of different countries. Most notably, stocks of Patagonian toothfish in Chile accumulate significantly more mercury than stocks closer to the South Pole, including the South Georgia/Shag Rocks stock, a fishery certified by the Marine Stewardship Council (MSC) as sustainably fished. Consistent with the documented geography of mercury contamination, our analysis showed that, on average, retail fish labeled as MSC-certified Patagonian toothfish had only half the mercury of uncertified fish. However, consideration of genetic data that were informative about seafood substitutions revealed a complex pattern of contamination hidden from consumers: species substitutions artificially inflated the expected difference in mercury levels between MSC-certified and uncertified fish whereas fishery stock substitutions artificially reduced the expected difference in mercury content between MSC-certified and uncertified fish that were actually D. eleginoides. Among MSC-certified fish that were actually D. eleginoides, several with exogenous mtDNA haplotypes (i.e., not known from the certified fishery) had mercury concentrations on par with uncertified fish from Chile. Overall, our analysis of mercury was consistent with inferences from the genetic data about the geographic origins of the fish, demonstrated the potential negative impact of seafood mislabeling on unwanted mercury exposure for consumers, and showed that fishery-stock substitutions may expose consumers to significantly greater mercury concentrations in retail-acquired fish than species substitutions.

Fatty acid composition and dynamics of phospholipids from hake (Merluccius hubbsi) spinal cord and brain and sea bass (Acanthustius brasilianus) brain.

This work studies the phospholipid and fatty acid composition in hake brain and spinal cord and in sea bass brain. Fluorescence anisotropy of phospholipid vesicles labeled with 1,6-diphenyl hexatriene was measured to investigate the associated dynamic properties. In all tissues studied, phosphatidylcholine and phosphatidylethanolamine were the major constituents with minor contributions of phosphatidylserine, phosphatidylinositol and sphingomyelin. Fatty acids belong to the n-9 and n-3 series exclusively. Phosphatidylinositol from hake spinal cord and phosphatidylethanolamine and phosphatidylserine from hake brain contain the greatest percentages of eicosa-5,8,11,14,17-pentaenoic (20:5) and docosa-4,7,10,13,16,19-hexaenoic (22:6), respectively. For all fractions studied the total content of saturated fatty acids increases in the order of hake spinal cord, hake brain, sea bass brain together with a decrease in the sum of monounsaturated fatty acids. The comparison between fluorescence anisotropy values and fatty acid composition clearly demonstrates that saturated acids and 20:5 and 22:6 exert a rigidizing effect.