Per- and polyfluoroalkyl substances in Chinese commercially available red swamp crayfish (Procambarus clarkii): Implications for human exposure and health risk assessment.

The extensive utilization of per- and polyfluoroalkyl substances (PFASs) has led to their pervasive presence in the environment, resulting in contamination of aquatic products. Prolonged exposure to PFASs has been linked to direct hepatic and renal damage, along with the induction of oxidative stress, contributing to a spectrum of chronic ailments. Despite the recent surge in popularity of red swamp crayfish as a culinary delicacy in China, studies addressing PFASs' exposure and associated health risks from their consumption remain scarce. To address this gap, our study investigated the PFASs' content in 85 paired edible tissue samples sourced from the five primary red swamp crayfish breeding provinces in China. The health risks associated with dietary exposure were also assessed. Our findings revealed widespread detection of PFASs in crayfish samples, with short-chain perfluoroalkyl carboxylic acids (PFCAs) exhibiting the highest concentrations. Notably, the total PFAS concentration in the hepatopancreas (median: 160 ng/g) significantly exceeded that in muscle tissue (5.95 ng/g), as did the concentration of every single substance. The hazard quotient of perfluorohexanesulfonic acid (PFHxS) via consuming crayfish during peak season exceeded 1. In this case, a potential total non-cancer health risk of PFASs, which is mainly from the hepatopancreas and associated with PFHxS, is also observed (hazard index>1). Thus, it is recommended to avoid consuming the hepatopancreas of red swamp crayfish. Greater attention should be paid to governance technology innovation and regulatory measure strengthening for short-chain PFASs.

Elucidation of key fatty aroma compound contributing to the hepatopancreas of Eriocheir sinensis using sensomics approach by GC-IMS and GC-MS-O.

Aroma is one of the most noticeable characteristics when consuming Chinese mitten crab (Eriocheir sinensis) and is crucial for consumer satisfaction and the development of industry. In this study, we utilized fingerprints and the sensomics approach to analyze volatiles in the hepatopancreas of E. sinensis from Chongming and Taixing. GC-IMS indicated that the odor profile was dominated by pungent (-), buttery (+), and fruity (+) from Chongming and was more prone to alcoholic (-), solvent (-), and aldehydic (+) in Taixing. Moreover, PLS-DA modeling identified 2-acetylthiazole and toluene as the primary differential compounds. Subsequently, fifteen active-aroma compounds with FD values of >4 was recombined in an odorless matrix to simulate the odor profile of the hepatopancreas. Notably, removing methional may significantly decrease the intensity of the fatty and toasted odors. The findings reveal the odor profile of hepatopancreas and establish a theoretical foundation for subsequent studies on flavor.

Isolation, characterization and antimicrobial properties of hepatopancreas lectin of the freshwater crab Oziotelphusanaga.

Lectins are versatile proteins that specifically recognize and interact with sugar moieties expressed on the cell surface. The potential of lectin in drug targeting and delivery has instigated interest to identify natural lectins. Crabs have been identified as a rich source of lectin because the innate immune system is activated on encounter of pathogens and helps in the production of lectin. Although the presence of lectins in crab's hemolymph is well documented, little information about lectin in hepatopancreas, a vital organ for immunity and digestion in crustaceans, is currently available. A calcium dependent lectin (75 kDa) was purified from the hepatopancreas of the freshwater crab Oziotelphusa naga by bioadsorption and fetuin linked Sepharose 4B affinity chromatography technique. The isolated hepatopancreas lectin is calcium dependent and maximum agglutination was observed with rabbit erythrocytes. The hemagglutinating activity of the hepatopancreas lectin was effectively inhibited by sugars, such as α-lactose, GlcNAc, trehalose and NeuAc. Compared to sialylated N-glycosylated proteins including transferrin and apo transferrin, sialylated O-glycosylated proteins like fetuin exhibited stronger inhibitory effect. The ability of erythrocytes to bind hepatopancreas lectin has been diminished by desialylation of the potent inhibitor, indicating the significance of sialic acid in lectin-ligand interactions. The purified hepatopancreas lectin showed a broad spectrum of antimicrobial activity against bacteria Staphylococcus aureus, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, E. coli and fungi Candida albicans and Aspergillus niger. The findings of this study demonstrate the significance of hepatopancreas lectin as a multifunctional defense protein that inhibits the growth of bacteria and fungi.

Evaluation of Alternative Sources of Proteins and Other Nutrients with Potential Applications in Fish Nutrition.

The European Union's (EU) agricultural self-sufficiency is challenged by its reliance on imported plant proteins, particularly soy from the Americas, contributing to deforestation and greenhouse gas emissions. Addressing the EU's protein deficit, this study evaluates alternative protein sources for aquaculture, focusing on their nutritional value, elemental content, and polycyclic aromatic hydrocarbons (PAHs). Protein flours from gastropods (Helix pomatia, Arion lusitanicus, Arion vulgaris) and their hepatopancreas, along with plant-based proteins from food industry by-products (oilcakes, coffee grounds, spent brewer's yeast), were analyzed. Results revealed that snail flour contained the highest protein content at 59.09%, significantly outperforming hepatopancreas flour at 42.26%. Plant-based proteins demonstrated substantial nutritional value, with coffee grounds flour exhibiting a remarkable protein content of 71.8% and spent brewer's yeast flour at 57.9%. Elemental analysis indicated high levels of essential minerals such as magnesium in hepatopancreas flour (5719.10 mg/kg) and calcium in slug flour (48,640.11 mg/kg). However, cadmium levels in hepatopancreas flour (11.45 mg/kg) necessitate caution due to potential health risks. PAH concentrations were low across all samples, with the highest total PAH content observed in hepatopancreas flour at 0.0353 µg/kg, suggesting minimal risk of PAH-related toxicity. The analysis of plant-based protein sources, particularly oilcakes derived from sunflower, hemp, flax, and pumpkin seeds, revealed that these by-products not only exhibit high protein contents but present a promising avenue for enhancing the nutritional quality of feed. This study underscores the potential of utilizing gastropod and plant-based by-products as sustainable and nutritionally adequate alternatives to conventional feeds in aquaculture, contributing to the EU's environmental sustainability goals.

Concentrations, sources and health risk of bisphenols in red swamp crayfish (Procambarus clarkii) from South-Eastern China.

Bisphenol analogs (BPs) are extensively employed in commercial and industrial products and they have been found in a variety of environmental matrices and human samples. The red swamp crayfish (Procambarus clarkii) has been a trendy food in China in recent decades. However, the levels of BPs in Chinese crayfish and the associated hazards of human exposure remain unknown. Thus, in this study, the levels of eight BPs in crayfish gathered from five major provinces engaged in crayfish within the Yangtze River Basin were analyzed. Additionally, the health risks for humans by ingesting crayfish were calculated. BPs were frequently detected in crayfish tissues, indicating the wide occurrence of these chemicals. In comparison to other substitutions, BPA remains the dominant bisphenol analog. Most of the BPs were observed to accumulate in the hepatopancreas compared to the muscle, so consuming the hepatopancreas of crayfish is not recommended. With the exception of BPS, the Estimated Daily Intakes (EDIs) of the remaining BPs exceeded the Tolerable Daily Intake (TDI) specified by the European Food Safety Authority (EFSA) by a factor of 1.75-69.0. The mean hazard index (HI) values exceeded 1 for both hepatopancreas and muscle in all provinces, and the mean HI values for hepatopancreas were significantly higher than those for muscle, indicating potential health risks for local consumers.

Geographical traceability of flavor compounds in Chinese mitten crab (Eriocheir sinensis): Implications for quality differentiation, authenticity assessment, and mechanism research.

Geographical traceability plays a crucial role in ensuring quality assurance, brand establishment, and the sustainable development of the crab industry. In this study, we examined the possibility of using gas chromatography-ion mobility spectrometry with multivariate statistical authenticity analysis to identify the origin of crabs from five sites downstream of the Yangtze River. Significant variations were observed in the levels of alcoholic flavor compounds in the hepatopancreas and muscles of crabs from different geographical locations, and a support vector machine exhibited discriminant ability with 100% accuracy. These flavor variations exhibited significant correlations with the types and concentrations of elements within the crabs, as well as with free amino acids. This study offers a practical approach for determining the geographical traceability of Chinese mitten crabs and elucidates the role of elements in flavor modulation, thereby providing innovative strategies to enhance the efficiency of crab farming.

Exposure to virgin and marine incubated microparticles of biodegradable and conventional polymers modulates the hepatopancreas transcriptome of Mytilus galloprovincialis.

Biodegradable polymers have been proposed as an alternative to conventional plastics to mitigate the impact of marine litter, but the research investigating their toxicity is still in its infancy. This study evaluates the potential ecotoxicological effects of both virgin and marine-incubated microparticles (MPs), at environmentally relevant concentration (0.1 mg/l), made of different biodegradable polymers (Polycaprolactone, Mater-Bi, cellulose) and conventional polymers (Polyethylene) on Mytilus galloprovincialis by using transcriptomics. This approach is increasingly being used to assess the effects of pollutants on organisms, obtaining data on numerous biological pathways simultaneously. Whole hepatopancreas de novo transcriptome sequencing was performed, individuating 972 genes differentially expressed across experimental groups compared to the control. Through the comparative transcriptomic profiling emerges that the preponderant effect is attributable to the marine incubation of MPs, especially for incubated polycaprolactone (731 DEGs). Mater-Bi and cellulose alter the smallest number of genes and biological processes in the mussel hepatopancreas. All microparticles, regardless of their polymeric composition, dysregulated innate immunity, and fatty acid metabolism biological processes. These findings highlight the necessity of considering the interactions of MPs with the environmental factors in the marine ecosystem when performing ecotoxicological evaluations. The results obtained contribute to fill current knowledge gaps regarding the potential environmental impacts of biodegradable polymers.

Comparison of Cd, Cu, Se, and Zn Concentration in the Muscle and Hepatopancreas of Sepia pharaonis and Uroteuthis duvauceli in the North of Persian Gulf (Iran).

The objective of this research was to examine and contrast the levels of cadmium (Cd), copper (Cu), zinc (Zn), and selenium (Se) in the muscle and hepatopancreas tissues of two species, namely pharaoh cuttlefish (Sepia pharaonis) and Indian squid (Uroteuthis duvauceli), from the Persian Gulf. A total of thirty individuals of each species were gathered in January 2009 from the northern waters of the Persian Gulf. The metal concentrations were significantly higher in muscle tissue (p < 0.05) than in other tissues. S. pharaonis had higher metal concentrations than U. duvauceli. In the muscle and hepatopancreas samples of S. pharaonis, the highest mean concentrations were found to be for Zn (58.45 ± 0.96 µg/g dw) and Cu (1541.47 ± 192.15 µg/g dw), respectively. In U. duvauceli, the highest concentration of measured elements was seen for Zn in both muscle (36.52 ± 0.56 µg/g dw) and hepatopancreas (60.94 ± 2.65 µg/g dw). Se had the lowest concentration among the elements measured in both species. There was a negative and significant correlation between Cu and biometrical factors (total body length and weight) in both muscle and hepatopancreas samples of S. pharaonic and only in the muscle samples of U. duvauceli (p < 0.01, R2 = - 052; p < 0.01, R2 = - 0.055). However, there was a strong correlation between Zn and biometrical factors in hepatopancreas samples of both species. The comparison of metal concentrations with standards revealed that only Cd levels in S. pharaonis exceeded the ESFA and WHO standards, whereas other metals were below the standards.

Waterless live transport degrades the flesh quality of Litopenaeus vannamei by disturbing neuroendocrine response: based on physiology and metabolomics.

BACKGROUND: Shrimp is one of the most popular marine foods consumed throughout the world and its freshness is a crucial indicator for consumers. However, the flesh quality degradation of shrimp during waterless live transport has been observed and the underlying mechanism remains unknown. RESULTS: The present study aimed to clarify the biochemistry mechanisms of flesh degradation with integration of quality evaluation, metabolic profiling and histopathological analysis. The flesh quality indicators such as water holding capacity, protein and lipid contents, amino acid composition and myofiber components degraded with the prolongation of combined stress. In addition, the metabolites including gamma-aminobutyric acid, Val-Ala, Trh and derivatives of carnitine, phosphocholine and prostaglandin all reduced significantly under combined stress (P < 0.05). Furthermore, Kyoto Encyclopedia of Genes and Genomes (https://www.genome.jp/kegg) analysis revealed the enrichment of neuroactive ligand-receptor interaction and estrogen signaling pathways, indicating the involvement of neuroendocrine in stress response. Moreover, architecture impairment in hepatopancreas tissue verified the accumulation of metabolic disturbance. CONCLUSION: Taken together, the findings of the present study indicate that neuroendocrine system mediates the flesh degradation of L. vannamei during waterless transport by disturbing the biochemical metabolic pathways and inducing architecture impairment of myofibril components. © 2022 Society of Chemical Industry.

Metabolomics of the hepatopancreas in Chinese mitten crabs (Eriocheir sinensis).

Chinese mitten crab (Eriocheir sinensis) is an important economic seafood in China, and the hepatopancreatic dim color and bitter taste of crabs significantly affects the consumer's acceptance. The crabs with brown hepatopancreas (BH) generally taste bitter, while the orange hepatopancreas (OH) does not taste bitter. While the metabolites perform their physiological functions, their metabolic relationship may directly affect the color and taste quality of the crab hepatopancreas. This study investigated the metabolic relationship of key metabolites related to the color and bitterness in the OH and BH of E. sinensis. The contents of total bile acids (BAs) (10.62 vs. 3.72 µg/g), bitter free amino acids (FAAs) (5.15 vs. 2.46 mg/g) and lutein (6.88 vs. 2.55 µg/g) in the BH group were significantly higher (P < 0.01) than those in the OH group, while ß-carotene (314.47 vs. 478.52 µg/g) in the BH group was significantly lower (P < 0.05) than that in the OH group. In addition, BAs were positively correlated with lutein, saturated and monounsaturated fatty acids (SFA and MUFA) and bitter FAAs, and negatively correlated with color value, ß-carotene and polyunsaturated fatty acids (PUFA). In conclusion, BAs and bitter FAAs may be the major contributors to the bitterness of BH, and the high content of BAs in the BH group may promote lipid catabolism, inhibit ß-carotene absorption and enhance amino acid metabolism, leading to the hepatopancreas brown and bitter.

Fluoride sensitivity in freshwater snail, Bellamya bengalensis (Lamarck, 1882): An integrative biomarker response assessment of behavioral indices, oxygen consumption, haemocyte and tissue protein levels under environmentally relevant exposure concentrations.

There is limited information on fluoride toxicity and risk overview on ecotoxicological risks to aquatic invertebrate populations particularly molluscan taxa. This necessitated the assessment of toxicity responses in the freshwater snail, Bellamya bengalensis exposed to environmentally relevant concentrations of sodium fluoride. Under lethal exposures (150, 200, 250, 300, 400 and 450 mg/l), the median lethal concentrations (LC50) were determined to be 422.36, 347.10, 333.33 and 273.24 mg/l for B. bengalensis at 24, 48, 72 and 96 h respectively. The rate of mortality of the snails was increased significantly with elevated concentrations of the toxicant. The magnitude of toxicity i.e., toxicity factor at different time scale was also higher with increased exposure duration. Altered behavioural changes i.e., crawling movement, tentacle movement, clumping tendency, touch reflex and mucous secretion in exposed snail with elevated concentrations and exposure duration. Similarly, oxygen consumption rate of the treated snail also lowered significantly during 72 and 96 h of exposure. Under 30-day chronic exposures (Control-0.00 mg/L; T1-27.324 mg/L; T2-54.648 mg/L), protein concentrations in gonad and hepatopancreas of exposure groups was significantly lowered. Chronic exposures also revealed lowered haemocytes counts in exposure groups. The potential for loss of coordination, respiratory distress and physiological disruption in organisms exposed to environmentally relevant concentrations of fluoride was demonstrated by this study. The estimation and magnitude of toxicity responses are necessary for a more accurate estimation of ecological risks to molluscan taxa and invertebrate populations under acute and chronic fluoride exposures in the wild.

Probiotic supplementation affects IGF-1 and leptin levels in Nile tilapia hepatopancreatic tissue / Suplementação com probiótico afeta os níveis de IGF-1 e leptina no tecido hepatopancreático de tilápias-do-nilo

This work aimed to assess the effect of the probiotic strain, Lactobacillus plantarum, on the levels of leptin, IGF-1 and their receptors on the hepatopancreatic tissues of Nile tilapia (Oreochromis niloticus) and then correlate fish growth performance and gut microbiological parameters. Fish juveniles (±23g) were reared in a recirculation system with constant aeration and temperature (25°C). They were distributed into six polyethylene tanks (45L) and fed twice a day at 5% of the tank biomass with the respective diets: control (commercial diet without probiotic) and supplemented with L. plantarum inoculum (1 x 108 CFU mL-1), both in triplicate. After 30 days of feeding, L. plantarum-fed fishes showed greater weekly growth rate, final weight, and feed conversion rate, in addition to higher count of lactic-acid bacteria and lower count of pathogenic bacteria in the intestinal tract, when compared to the control group. The immunostaining intensity for IGF-1 and leptin hormones was lower after L. plantarum supplementation than in the control group, with no change in the level for receptors. This reduction could implicate important changes in fish metabolism and homeostasis.(AU)

O presente trabalho teve como objetivo avaliar o efeito da cepa probiótica Lactobacillus plantarum sobre os níveis de leptina, IGF-1 e seus receptores no tecido hepatopancreático de tilápia-do-nilo (Oreochromis niloticus) e correlacionar com o desempenho zootécnico e os parâmetros microbiológicos intestinais dos peixes. Juvenis de tilápia-do-nilo (±23g) foram distribuídos em seis tanques de polietileno (45L) conectados a um sistema de recirculação, com aeração e temperatura constantes (25°C). Os peixes foram alimentados duas vezes ao dia, a 5% da biomassa do tanque, com as respectivas dietas: controle (dieta comercial sem probiótico) e suplementada com L. plantarum (1 x 108 UFC mL-1), ambas em triplicata. Após 30 dias de cultivo, os peixes alimentados com L. plantarum apresentaram maiores ganho de peso semanal, peso final e conversão alimentar, bem como maior contagem de bactérias ácido-láticas e menor contagem de bactérias patogênicas no trato intestinal das tilápias alimentadas com dieta probiótica, em comparação ao grupo controle. A intensidade da imunomarcação para os hormônios IGF-1 e leptina foi menor com a suplementação de L. plantarum do que no grupo controle, sem alterar os níveis de seus receptores. Essa redução pode implicar mudanças importantes no metabolismo e na homeostase dos peixes.(AU)

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A method for geographical discrimination of Portunus trituberculatus was explored to provide technical support for the protection of geographical indication products and for tracing the origin of seafood. P. trituberculatus were collected from three major production areas, including the Yellow Sea, the Bohai Sea, and the East China Sea. The variations of carbon and nitrogen stable isotope values of origins and the correlation of stable isotope ratios in different tissues were analyzed. The results showed that there were significant differences in carbon and nitrogen stable isotope ratio among different origins. Significant isotope fractionation effects were observed among different tissues. The discriminant model was developed and the origin discriminant analysis was performed by the stable isotope ratios of different tissues in P. trituberculatus. The correct rate of origin diffe-rentiationf using carbon and nitrogen stable isotopes in muscle and gills (>95%) was significantly higher than that of hepatopancreas and gonad, indicating that stable isotope ratios of muscle and gills could effectively differentiate P. trituberculatus in different sea areas. This study filled the gap of stable isotope tracing technology for P. trituberculatus.

Comparison on nutritional quality of adult female Chinese mitten crab (Eriocheir sinensis) with different colored hepatopancreases.

White hepatopancreas syndrome has recently emerged in Chinese mitten crab (Eriocheir sinensis) aquaculture, causing considerable economic loss. The hepatopancreas color of diseased crabs becomes gradually lighter, turning from yellow to yellow-white to white. Therefore, this study was conducted to investigate the changes in nutrient composition in three edible parts (hepatopancreas, ovaries, and muscle) of adult females with different colored hepatopancreases. Three groups were assessed in this study, including a yellow hepatopancreas group (control, L * = 63.92, a * = 22.14, b * = 60.95), a yellow-white hepatopancreas group (YWHG, L * = 65.06, a * = 22.35, b * = 57.80), and a white hepatopancreas group (WHG, L * = 65.72, a * = 10.70, b * = 30.52). No statistically significant differences in average weight, tissue indices, and total edible yield were observed among the three crab groups (P >0.05). The moisture content of the hepatopancreases and ovaries in WHG was 56.12% and 9.23% higher than the control values (P <0.05), whereas hepatopancreas crude fat and ovary crude protein levels were 62.23% and 11.45% lower than the control values (P < 0.05). The total carbohydrate levels of the three edible tissues were significantly higher and the crude protein content of ovaries was significantly lower in YWHG (P < 0.05). Most amino acid levels in the WHG muscle and ovaries were significantly lower than the control (P < 0.05). Moreover, the hepatopancreas levels of total polyunsaturated fatty acids (PUFA) and n-6PUFA in WHG were 24.88% and 31.83% lower than in control group, whereas the hepatopancreas levels of total PUFA and n-6PUFA in YWHG were also 21.88% and 23.20% lower compared to the controls (P < 0.05). Overall, the growth and the edible parts were not affected in YWHG and WHG. Moreover, YWHG crabs exhibited few effects on nutritional value; however, the fatty acid composition of crabs was significantly changed. In contrast, WHG crabs exhibited poor nutritional quality. Nonetheless, the consumption of crabs with yellow-white or white hepatopancreases is not recommended since the animal also referred to as diseased crabs.

Large lipid transfer proteins in hepatopancreas of the mud crab Scylla paramamosain.

Large lipid transfer proteins (LLTPs) are extensively involved in various physiological processes. In the present study, five LLTP sequences encoding apolipocrustacein 1 (apoCr 1), apoCr 2, precursor of the large discoidal lipoprotein (dLp) and high density lipoprotein/ß-glucan binding protein (HDL-BGBP) (dLp-BGBP), microsomal triglyceride transfer protein (MTP) and clotting protein (CP) were identified in the hepatopancreas of Scylla paramamosain. Of these, apoCr 2, dLp-BGBP, and MTP were newly identified in this species, and the former two proteins were classified into the APO family while the later into the MTP family in phylogenetic trees. The apoCr 1 expression level was dramatically increased in the hepatopancreas towards ovarian maturation, which was extremely greater than that in the ovaries concurrently, likely to meet the considerable requirements of yolk protein and lipids for embryo development. The dLp-BGBP expression level in male crabs was comparable to that in female crabs, supporting HDL-BGBP acts as a major circulatory lipid carrier. The close phylogenetic relationship between dLp-BGBP and the scaffolding protein of lipid transfer particle implied dLp might facilitate lipid transfer between the hepatopancreas and HDL-BGBP-containing lipoproteins. The MTP expression level was positively related to ovarian development in both the hepatopancreas and ovaries, indicating MTP may be involved in lipoprotein assembly in the hepatopancreas and lipid droplet maturation in the ovaries. CP may play a crucial role in embryo development based on high expression level observed in the testes of mature crabs. Our findings provide novel insights into LLTP superfamily members and their functions in decapods.

Lobster Supply Chains Are Not at Risk from Paralytic Shellfish Toxin Accumulation during Wet Storage.

Lobster species can accumulate paralytic shellfish toxins (PST) in their hepatopancreas following the consumption of toxic prey. The Southern Rock Lobster (SRL), Jasus edwardsii, industry in Tasmania, Australia, and New Zealand, collectively valued at AUD 365 M, actively manages PST risk based on toxin monitoring of lobsters in coastal waters. The SRL supply chain predominantly provides live lobsters, which includes wet holding in fishing vessels, sea-cages, or processing facilities for periods of up to several months. Survival, quality, and safety of this largely exported high-value product is a major consideration for the industry. In a controlled experiment, SRL were exposed to highly toxic cultures of Alexandrium catenella at field relevant concentrations (2 × 105 cells L-1) in an experimental aquaculture facility over a period of 21 days. While significant PST accumulation in the lobster hepatopancreas has been reported in parallel experiments feeding lobsters with toxic mussels, no PST toxin accumulated in this experiment from exposure to toxic algal cells, and no negative impact on lobster health was observed as assessed via a wide range of behavioural, immunological, and physiological measures. We conclude that there is no risk of PST accumulation, nor risk to survival or quality at the point of consumption through exposure to toxic algal cells.

Comparison of growth performance and biochemical components between parent and hybrid offspring in the oriental river prawn, Macrobrachium nipponense.

Macrobrachium nipponense, as one of the large-yield farmed shrimp, is facing germplasm degradation. Genetic improvement through hybridization is one of the effective methods to solve this problem. In this study, using a three-line hybrid strategy, two-hybrid F1 populations were obtained using three local populations of M. nipponense as parents for crossbreeding. Five populations were then cultured for 3 months. Growth rate performance was measured by the hepatosomatic index, weight gain, body length growth rate and special growth rate. Biochemical components were also assessed. The results showed that the survival rate and growth performance of the hybrid progeny were better than those of the parents. The levels of triglycerides, total cholesterol, glycogen and lactic acid of the hybrid population were higher than those of the parents. This was consistent with variation in the activity of four digestive enzymes. Compared with the results of the fatty acid and amino acid analysis, it was found that the contents of polyunsaturated fatty acids such as eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid and eight essential amino acids in the hybrid populations were significantly higher than those of their parents, and the contents of flavor amino acids were higher. The expression level of molting genes related to the growth of the parent populations was lower than that of the hybrids. These results show that crossbreeding is effective for the genetic improvement of M. nipponense germplasm. Hybrids showed advantages in growth and nutrition and multigenerational breeding will be required to form a stable germplasm.

Hypoxia-reoxygenation stress modulates the hepatopancreas transcriptome of Chinese mitten crab Eriocheir sinensis.

Hypoxia is a critical, but frequently overlooked problem, which commonly exists in Chinese mitten crab rearing. However, little information is available on the molecular mechanisms of the detrimental effects of hypoxia in this species. In the present study, crabs were subjected to acute hypoxia stress (DO 1.0 mg/L), followed by reoxygenation (DO 6.8 mg/L). Hepatopancreas from five groups of crabs (three to four crabs per group), including normoxia, hypoxia for one and six hours, and reoxygenation for one and 12 h, were used for transcriptome sequencing. The pooled total RNA of all samples were utilized to reconstruct a reference transcriptome with PacBio RS II sequencing, obtaining 49.19 G clean data, with a mean length of 2,180 bp. Seventeen cDNA libraries were constructed and sequenced to identify differentially expressed genes (DEGs) among the different samples (FDR < 0.05 and |log2 fold change| ≥1). A total of 103 and 251 DEGs were identified when exposed to hypoxia for one and six hours, respectively. Totally 462 and 673 DEGs were identified during reoxygenation at 1 and 12 h, respectively. Among these DEGs, two transcripts with complete ORFs were identified to encode hypoxia-inducible factor 1 (Es-Hif-1α/ß), which is a transcriptional activator of various genes correlated to the cellular adaptive responses to hypoxia. Es-Hif-1a/ß expressions were significantly upregulated when exposed to hypoxia treatment, and no significant difference was observed for Es-Hif-1α between hypoxic treatment for 6 h and reoxygenation. The significant KEGG enrichment revealed that the DEGs under hypoxia were mainly enriched in "PPAR signaling pathway", "Gap junction" and "Phototransduction-fly". Compared with crabs in normoxia, even with 12 h of reoxygenation, the hepatopancreas transcriptome under hypoxia remained severely affected, implying its adverse effect on growth and development, or increased susceptibility to diseases.

Recycling biofloc waste as novel protein source for crayfish with special reference to crayfish nutritional standards and growth trajectory.

Screening of novel feedstuffs, that too for data-deficient (nutritionally) animals, is somewhat ambiguous or problematic. Through systematic meta-analyses, the present study formulated most up-to-date crayfish nutritional standards, against which a recyclable waste (biofloc biomass, BM) from intensive aquaculture systems was assessed as a novel protein source. Growth trajectory dependencies and thermal growth coefficient qualifying for good growth in crayfish (TGC 0.5-0.64 units) were benchmarked. Using these standards and a 7-week growth trial, BM's suitability as a novel protein source for red swamp crayfish Procambarus clarkii was evaluated through its graded inclusions in a commercial feed. Results suggest that BM can elevate growth at 33-66% inclusion in existing feed formulations. Beyond 66% inclusion, BM can deteriorate growth in crayfish due to high ash content (exceeding physiological limit > 14%), arginine deficiency (~ 14-20% lower than an optimum requirement), and insufficient non-protein energy: protein ratio (3.7 cal mg-1). Arginine is perhaps the most critical amino acid in dietary protein for crayfish, and deficient in BM. Although no critical bioaccumulation levels of heavy metals were breached by feeding 100% BM to crayfish, a mineral and heavy metal (Hg) stress seemed plausible. Crayfish raised solely on biofloc may not realize full growth potential.

Comparative assessment of biomarker response to tissue metal concentrations in urban populations of the land snail Helix pomatia (Pulmonata: Helicidae).

The traffic pressure is increasing, resulting in the emission of atmospheric pollution. Soil organisms will need to respond to pollution stressors. Among them, land snails are valuable indicators of ecosystem disturbance. In this study, land snails Helix pomatia were sampled from three city localities with different traffic intensity. Oxidative stress biomarkers catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione-S-transferase (GST) in the foot muscle (FM) and hepatopancreas (HP) tissue were determined. Also, five heavy metal (Cd, Cu, Ni, Pb, and Zn) concentrations were quantified in soil and tissue samples. According to the results, the highway induces the strongest contamination on the surrounding environment, with the highest metal concentrations measured in soil and snails. At the most polluted locality, only Cd exceeded some soil guidelines authorities that we referred to in this study. In addition, tissue Cd concentrations exceeded the United States Environmental Protection Agency (USEPA) value (1 mg kg-1) for soil invertebrate toxicity at all localities making it likely responsible for generating adverse effects in snails. Regarding HP, the CAT and GST are the most sensitive parameters that could be useful as oxidative stress biomarkers in snails exposed to the actual metals in the environment. On the other hand, in FM tissue, the most pronounced changes were recorded for GPX and GR. Based on tissue-specific enzyme responses, three urban populations were clearly separated. Therefore land snails are the promising candidates for quick field-based biomarker studies after showing a tissue-specific concentration-dependent induction of certain enzymes to heavy metals.