Search

In an era when ecological and environmental needs and responsibilities apply pressure on the world's countries and sustainability takes centre stage, ecologic/environmental (E/E) laboratories stand as beacons of scientific inquiry, innovating, optimising, and applying various tests for a better knowledge of our natural resources and the quality status of ecosystems. The purpose of this review is to provide an overview of the use of flow cytometry (FC) as a tool for assessing environmental quality, mainly using living organisms and their biological changes as bioindicators. Cytometric approaches applied to both marine and terrestrial ecosystems ensure the detection of biochemical and functional status of the cells composing either an organ thereof or the organism itself. In addition to cytometric evaluations of the biotic matrix, a brief overview of the techniques for the environmental assessment of biotic and abiotic matrices using mass spectrometry is given. The technique involving the continuous monitoring of the chemical and physical parameters of water, sediment, and soil is basically incapable of detecting any additive and synergetic effects of toxicants on living organisms. Therefore, techniques employing bioindicators provide valuable information for environmental diagnosis, and several studies have demonstrated the strong relationship between specific environmental data and cell/organ behaviour. [ABSTRACT FROM AUTHOR]

The toxic effect and differences of Cr(VI) on superoxide dismutase (SOD) activity, MDA, and metallothionein (MT) content in the hepatopancreas and gill of Portunus trituberculatus were investigated during Cr(VI) enrichment (15 days) and release experiments (15 days). Results showed that the 1.50 and 0.30 mg/L test groups significantly exhibited higher SOD, MDA, and MT content in the hepatopancreas and gill compared with the control group after 15 days of enrichment (p < 0.05). After 15 days of Cr release, the SOD, MDA, and MT content in the hepatopancreas and gill of both test groups recovered to the normal level of the control group (p > 0.05). The gill of P. trituberculatus achieved the highest SOD activity, MDA, and MT content earlier than the hepatopancreas, but the highest values were lower. The gill showed a shorter recovery time than the hepatopancreas. We concluded that the gill of P. trituberculatus exhibited a more rapid response to, and recovery from, Cr(VI) exposure compared to the hepatopancreas, making it a more sensitive tissue for assessing Cr(VI) toxicity, though both tissues showed a capacity for recovery after the removal of the contaminant. [ABSTRACT FROM AUTHOR]

Human enteric viruses, such as hepatitis A virus (HAV), hepatitis E virus (HEV), and norovirus genogroups I and II (NoVGI and NoVGII), cause infections, and it has been largely demonstrated that mussels play an important role if consumed as raw or undercooked food matrices. This study aimed to investigate, through qualitative and quantitative biomolecular assays, the detection of partial genomic regions belonging to the most relevant enteropathogenic viruses for humans (HAV, HEV, NoVGI and NoVGII) in mussels (Mytilus galloprovincialis) farmed along the coasts of two Italian regions on the central Adriatic Sea: Abruzzo (Casalbordino, Chieti) and Molise (Termoli, Campobasso). A total of 425 animals were sampled, and the respective georeferentiations were registered. A total of 85 pools, each composed of five subjects/aliquots, were formed (22 from Abruzzo and 63 from Molise regions). This step was followed by homogenization and RNA extraction, and then the biomolecular assays [nested reverse transcription polymerase chain reaction (PCR) and real-time reverse transcription-quantitative PCR] were performed. 1.17% of the pool was positive for HAV RNA detection (10² copies/mL), 9.41% for HEV (10²-10³ copies/µL), 2.35% for NoVGI (10¹ copies/µL), and no pool was positive for NoVGII. This study demonstrated the human enteric viruses' presence in mussels farmed in a low-investigated marine area. Based on a one-health point of view, this paper aims to enforce the importance of biomolecular and epidemiological screenings as surveillance systems to guarantee human, animal, and environmental health. [ABSTRACT FROM AUTHOR]

Simple Summary: The oriental river prawn, Macrobrachium nipponense, is a widely distributed species in freshwater and low-salinity estuarine regions of China and other Asian countries. This species has become an important commercial commodity in China because of its high nutritional value and palatability. There are significant differences in growth between male and female M. nipponense. The aim of this study was to determine the hepatopancreas transcriptome differences between sex-related size differences in M. nipponense. We identified four genes associated with sex-related size differences, as well as six closely related metabolic pathways. The results indicated the molecular mechanism underlying the sex-related size differences and identified key genes and metabolic pathways. This data will be invaluable to support explanations of individual differences between male and female prawns. Macrobrachium nipponense, a commercially popular crustacean species within the Chinese context, is recognized for its exceptional nutritional composition and palatability. There are significant differences in growth between male and female M. nipponense. Herein, transcriptomics was used to determine the hepatopancreas transcriptome differences between sex-related size differences in M. nipponense. We identified 974 differentially expressed genes (DEGs) between the SHE (female) and BHE (male) groups, which were validated using RT-qPCR. The genes encoding matrix metalloproteinase-9 (MM9), Ribosome-binding protein 1 (RBP1), Aly/REF export factor 2, and hematological and neurological expressed 1 (HN1) may play a role in modulating the sex-related size differences observed in M. nipponense. Clusters of orthologous groups and gene ontology functional analysis demonstrated that the DEGs for sex-related size in M.nipponense were associated with various biological functions. The Kyoto Encyclopedia of Genes and Genomes pathways analysis demonstrated that upregulated DEGs were mainly enriched in lysine biosynthesis, tryptophan metabolism, and lysine degradation pathways, whereas the downregulated DEGs were mainly enriched in ascorbate and aldarate metabolism, retinol metabolism, and drug metabolism-cytochrome P450 pathways. The results indicated the molecular mechanism underlying the sex-related size differences and identified key genes. This data will be invaluable to support explanations of individual differences between male and female prawns. [ABSTRACT FROM AUTHOR]

Nitrite constitutes a significant pollutant that impedes the growth of P. vannamei. Conventional treatments are primarily geared toward decreasing nitrite concentrations from an environmental standpoint. Nevertheless, investigating methodologies to enhance the ability of Pacific white shrimp to withstand nitrite-induced stresses on an individual basis remains an unexplored topic. The present study examines the impact of autophagy on Pacific white shrimp's response to high nitrite environments through rapamycin injection. The research findings indicate that activating autophagy can effectively enhance the survival rate of Pacific white shrimp under high nitrite conditions. Additionally, total hemocyte count (THC) results in the hemolymph demonstrate that autophagy can alleviate the reduction in blood cell count caused by nitrite stress. The transcriptome results show that upregulation of 718 genes and downregulation of 1071 genes after nitrite stress. The majority of these differentially expressed genes (DEGs) were related to physiological processes, including oxidative stress, energy metabolism, and lysosomes. In addition, there were 911 upregulated genes and 713 downregulated genes upon activation of autophagy. These DEGs are associated with immune responses, specifically involving glutathione metabolism, drug metabolism-cytochrome P450, and metabolism of xenobiotics by cytochrome P450, lysosomes, autophagy, phagosomes, melanization, and MAPK signaling pathways. These pathways can reduce oxidative stress and promote the metabolism of harmful substances in the body, in shrimp, thereby improving their survival rates. These findings provide new insights for research aimed at enhancing nitrite tolerance in Pacific white shrimp. [ABSTRACT FROM AUTHOR]

Background: The Pacific white shrimp is one of the world’s most economically significant aquatic species, being one of the top three species cultured globally. However, the increasing incidence of diseases such as acute hepatopancreatic necrosis disease and hepatopancreatic microsporidia has led to a serious decline in shrimp production and severe economic losses. With the increasing demand for pathogen detection in shrimp farms, rapid DNA extraction technology has become more sophisticated. In this study, a rapid and crude method of extracting genomic DNA from shrimp muscle and hepatopancreas using Chelex-100 was established. Results: DNA was successfully extracted from muscle and hepatopancreatic tissues using both the Chelex-100 method and commercial kits. The internal reference genes of shrimp were successfully amplified via PCR and real-time PCR using the obtained DNA samples. Moreover, a field assay was successfully conducted using real-time PCR and real-time enzymatic recombinase amplification (real-time ERA), indicating that the quality of the DNA extracted using Chelex-100 is sufficient for use in conjunction with nucleic acid amplification to detect pathogens in shrimps. Conclusions: Chelex-100 is an efficient method for extracting DNA from shrimp muscle or hepatopancreas tissues, with a short extraction time, high extraction efficiency, and simple operation, making it appropriate for use in the detection of pathogens in shrimp.How to cite: Yang H, Zhou Q, Hu J, et al. A Chelex-100-based rapid DNA extraction method and its application in the detection of shrimp pathogens. Electron J Biotechnol 2024;70. https://doi.org/10.1016/j.ejbt.2024.04.004.

Biofloc systems represent a promising approach for achieving greater sustainability and productivity in aquaculture. This trial aimed to investigate the physio-histological impacts of different concentrations of biofloc on antioxidant capacities and the histology of both hepatopancreas and gills structure and abnormalities of the white leg shrimp (Litopenaeus vannamei). The study used three ratios of C:N (1:10, 1:15, 1:20) with three different levels of density (10, 20, 20 individuals/100L) for each ratio with triplicate for 56 days. The results of antioxidants activities showed that antioxidants (SOD, CAT, GPXs, MDH, LYZ) were significantly increased in biofloc treatments and reached its best activity in 1:15 treatment with a stocking density of 20 individuals/100L. The histological records indicated that 1:15 ratio enhanced the structure for both hepatopancreas and gills tissues. Tissues in the control and 1:10 ratio treatment seemed to be normal. In the 1:15 treatment, the hepatopancreas showed all important cells as blazeZellen cells (B) and embryonal Zellen cells (E) and normal arrangement of cells and normal interlamellar space for gills. With the increase of C:N ratio and stocking density, hepatopancreas showed a severe cell deformation, vacuoles, while the gills showed epithelial edema and abnormalities, particularly in 1:30 treatment, with 30 individuals/100L. [ABSTRACT FROM AUTHOR]

Phosphorus, calcium, and magnesium are pivotal elements in the exoskeleton of crustaceans, playing a crucial role in the moulting stages. As crustaceans cannot synthesize these nutrients, their acquisition relies on external sources, with tissues such as the hepatopancreas and gills actively participating in this process. Studying changes in phosphorus, calcium, and magnesium concentrations in these tissues is essential for a comprehensive understanding of their role in nutrient absorption during the moulting stages. The hepatopancreas, gills, muscles, and carapace of Macrobrachium nipponense (De Haan, 1849) were taken from the premoult, postmoult, and intermoult stages to study the changes in phosphorus, calcium and magnesium contents in the hepatopancreas, gills, muscles, and carapace during the moulting process of M. nipponense. Three parallel experiments were measured in each stage, and 10-20 prawns were mixed for each stage. Phosphorus content per tissue unit weight was measured using the vanadium-molybdenum yellow colorimetric method, and calcium and magnesium content per unit weight of tissue were measured using atomic absorption spectrophotometry. Phosphorus in the hepatopancreas and gills increased significantly from the premoult stage to the postmoult stage (P < 0.05), and magnesium also showed the same trend. Calcium in the hepatopancreas was significantly highest in the premoult phase, whereas phosphorus was significantly highest in the postmoult stage (P < 0.05). The hepatopancreas Ca/P ratio decreased significantly before and after moulting, while it increased significantly in the gills (P < 0.05). The results show that the hepatopancreas and gills are the main tissues that absorb phosphorus, calcium, and magnesium. The hepatopancreas absorbs and stores calcium earlier than phosphorus. The changes in the Ca/P ratio of the hepatopancreas and gills found that M. nipponense consumes more phosphorus than calcium during moulting and growth, and the gills have a better ability to absorb calcium than phosphorus. [ABSTRACT FROM AUTHOR]

A high-performance liquid chromatography method coupled to tandem mass spectrometry was validated in order to study the pharmacokinetics of cefotaxime in shrimp hepatopancreases and plasma, as well as its withdrawal time related to a maximum residue limit (MRL) in shrimp muscle. Pharmacokinetics parameters were investigated through oral medication at a single dose of 25 mg/kg shrimp body weight and subsequent hepatopancreas and plasma cefotaxime concentration measurements at 0.5, 1, 2, 4, 8, 12 and 24 h after shrimp were fed with medication. The maximum concentration of cefotaxime was observed after one hour in the hepatopancreas (Cmax, 19.45 ± 2.10 mg/kg) and 4 h in plasma (0.184 ± 0.061 mg/L). Based on a minimum inhibitory concentration (MIC) of cefotaxime of 4.13 mg/L against Vibrio parahaemolyticus (known to cause acute hepatopancreatic necrosis disease (AHPND) in white leg shrimp), it was observed that the time during which the hepatopancreas cefotaxime concentration was above the MIC was 23 h. An every 24 h cefotaxime treatment could thus be effective in fighting against this bacterium in shrimp. The withdrawal time of cefotaxime was determined after shrimp were fed with medicated feed once a day and twice a day for three consecutive days. Shrimp muscle was collected on day 1 and day 3 during medication and 1, 3, 7, 14 and 21 days after medication was stopped. Considering an MRL of 50 μg/kg, the withdrawal times were 8.5 degree-days (corresponding to 6.9 h at 29.5 °C) after shrimp were fed with medicated feed once a day for 3 days and 95.5 degree-days (77.7 h at 29.5 °C) after shrimp were fed with medicated feed twice a day for 3 days. Moreover, histological analysis revealed that feeding shrimp with cefotaxime at the given dose in once- or twice-a-day treatments did not negatively impact the shrimp hepatopancreas. [ABSTRACT FROM AUTHOR]

A lectin was isolated from the hepatopancreas of freshwater prawn, Macrobrachium rosenbergii by affinity chromatography using mucin-sepharose matrix. The purity of the isolated lectin was confirmed in native gradient PAGE that showed a single protein band of ∼37.9 kDa. In SDS-PAGE also one band of ∼43.3 kDa molecular weight was observed that indicated the protein to be a monomer. The band from the SDS-PAGE gel was identified through mass spectrometry as chitinase 1. The purified chitinase (50 μg/ml) hemagglutinated rabbit RBCs and, mucin and glucose inhibited hemagglutination with minimum concentrations of 0.1 mg/ml and 100 mM, respectively. Bacterial agglutination with Gram –ve Vibrio harveyi , Aeromonas sobria and Escherichia coli was also observed by this protein. Thus, chitinase 1 showed lectin-like properties besides its chitin hydrolytic activity. In western blot with hepatopancreas sample, rabbit antiserum against chitinase 1 cross-reacted to two additional proteins namely, chitinase 1C and obstructor E (a chitin-binding protein, CBP), besides its specific reactivity. An indirect ELISA was developed with the antiserum to quantify chitinases/CBP in hepatopancreas and serum samples of M. rosenbergii. The assay was used in samples from juvenile prawns following V. harveyi challenge. At 72 h post-challenge, significantly higher levels of chitinases/CBP were quantified in the hepatopancreas of the challenged group (1.8 ± 0.2 mg/g tissue) compared to the control (1.2 ± 0.1 mg/g tissue). This study suggests that the chitinase 1 protein with lectin-like properties is possibly induced at the protein level and can be putatively involved in the innate immune response of M. rosenbergii. • A ∼43.3 kDa Chitinase 1 was purified from the hepatopancreas of M. rosenbegii. • It possesses chitinolytic, hemagglutinating and bacterial agglutinating properties. • Antiserum to chitinase 1 cross-reacted to chitinase 1C and obstructor E proteins. • ELISA was developed to quantify chitinases/CBP in hepatopancreas and serum samples. • Vibrio harveyi challenge revealed chitinases/CBP to be inducible in nature. [ABSTRACT FROM AUTHOR]

Abstract Vibrio parahaemolyticus is a gram-negative facultative anaerobic bacterium implicated as the causative agent of several shrimp diseases. As part of the effort to provide biocontrol and cost-effective treatments, this research was designed to elucidate the effect of Morinda citrifolia fruit extract on the immunity of Penaeus vannamei postlarvae (PL) to V. parahaemolyticus. The methanol extract of M. citrifolia was vacuum evaporated, and the bioactive compounds were detected using gas chromatography‒mass spectrometry (GC‒MS). Thereafter, P. vannamei PL diets were supplemented with M. citrifolia at different concentrations (0, 10, 20, 30, 40, and 50 mg/g) and administered for 30 days before 24 h of exposure to the bacterium V. parahaemolyticus. A total of 45 bioactive compounds were detected in the methanol extract of M. citrifolia, with cyclononasiloxane and octadecamethyl being the most abundant. The survival of P. vannamei PLs fed the extract supplement was better than that of the control group (7.1–26.7% survival greater than that of the control group) following V. parahaemolyticus infection. Shrimp fed 50 mg/g M. citrifolia had the highest recorded survival. The activities of digestive and antioxidant enzymes as well as hepatopancreatic cells were significantly reduced, except for those of lipase and hepatopancreatic E-cells, which increased following challenge with V. parahaemolyticus. Histological assessment of the hepatopancreas cells revealed reduced cell degeneration following the administration of the plant extracts (expecially those fed 50 mg/g M. citrifolia) compared to that in the control group. Therefore, the enhanced immunity against V. parahaemolyticus infection in P. vannamei could be associated with the improved hepatopancreas health associated with M. citrifolia fruit extract supplementation.

Simple Summary: The pollution of aquatic ecosystems with the heavy metal copper (Cu) presents considerable risks to the physiological functions of aquatic organisms, including the economically valuable Macrobrachium rosenbergii. In this study, transcriptomic analysis was conducted to explore the molecular response mechanisms in the hepatopancreas of M. rosenbergii exposed to Cu. The results showed that M. rosenbergii attempts to resist the toxicity of Cu by up-regulating the expression of genes related to immunity, metabolism, and detoxification. However, with the excessive accumulation of reactive oxygen species (ROS), the antioxidant enzyme system was destroyed. As a result, DNA damage repair and the cellular stress response were inhibited, thereby exacerbating cell damage. In order to maintain the normal function of the hepatopancreas, M. rosenbergii removes damaged cells by activating the apoptosis mechanism. Differentially expressed genes (DEGs) and KEGG pathways identified in the present study not only facilitate an understanding of the molecular response mechanisms of M. rosenbergii underlying Cu toxicity effects but also help in identifying potential biomarkers associated with the stress response in other crustaceans. The contamination of aquatic ecosystems by the heavy metal copper (Cu) is an important environmental issue and poses significant risks to the physiological functions of aquatic organisms. Macrobrachium rosenbergii is one of the most important freshwater-cultured prawns in the world. The hepatopancreas of crustaceans is a key organ for immune defense, heavy metal accumulation, and detoxification, playing a pivotal role in toxicological research. However, research on the molecular response of the hepatopancreas in M. rosenbergii to Cu exposure is still lacking. In this study, the transcriptomic response in the hepatopancreas of M. rosenbergii was studied after Cu exposure for 3 and 48 h. Compared with the control group, 11,164 (7288 up-regulated and 3876 down-regulated genes) and 10,937 (6630 up-regulated and 4307 down-regulated genes) differentially expressed genes (DEGs) were identified after 3 and 48 h exposure, respectively. Most of these DEGs were up-regulated, implying that gene expressions were largely induced by Cu. Functional enrichment analysis of these DEGs revealed that immunity, copper homeostasis, detoxification, DNA damage repair, and apoptosis were differentially regulated by Cu. Seven genes involved in immunity, detoxification, and metabolism were selected for validation by qRT-PCR, and the results confirmed the reliability of RNA-Seq. All these findings suggest that M. rosenbergii attempts to resist the toxicity of Cu by up-regulating the expression of genes related to immunity, metabolism, and detoxification. However, with the excessive accumulation of reactive oxygen species (ROS), the antioxidant enzyme system was destroyed. As a result, DNA damage repair and the cellular stress response were inhibited, thereby exacerbating cell damage. In order to maintain the normal function of the hepatopancreas, M. rosenbergii removes damaged cells by activating the apoptosis mechanism. Our study not only facilitates an understanding of the molecular response mechanisms of M. rosenbergii underlying Cu toxicity effects but also helps us to identify potential biomarkers associated with the stress response in other crustaceans. [ABSTRACT FROM AUTHOR]

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Vibrio parahaemolyticus is a gram-negative facultative anaerobic bacterium implicated as the causative agent of several shrimp diseases. As part of the effort to provide biocontrol and cost-effective treatments, this research was designed to elucidate the effect of Morinda citrifolia fruit extract on the immunity of Penaeus vannamei postlarvae (PL) to V. parahaemolyticus. The methanol extract of M. citrifolia was vacuum evaporated, and the bioactive compounds were detected using gas chromatography‒mass spectrometry (GC‒MS). Thereafter, P. vannamei PL diets were supplemented with M. citrifolia at different concentrations (0, 10, 20, 30, 40, and 50 mg/g) and administered for 30 days before 24 h of exposure to the bacterium V. parahaemolyticus. A total of 45 bioactive compounds were detected in the methanol extract of M. citrifolia, with cyclononasiloxane and octadecamethyl being the most abundant. The survival of P. vannamei PLs fed the extract supplement was better than that of the control group (7.1–26.7% survival greater than that of the control group) following V. parahaemolyticus infection. Shrimp fed 50 mg/g M. citrifolia had the highest recorded survival. The activities of digestive and antioxidant enzymes as well as hepatopancreatic cells were significantly reduced, except for those of lipase and hepatopancreatic E-cells, which increased following challenge with V. parahaemolyticus. Histological assessment of the hepatopancreas cells revealed reduced cell degeneration following the administration of the plant extracts (expecially those fed 50 mg/g M. citrifolia) compared to that in the control group. Therefore, the enhanced immunity against V. parahaemolyticus infection in P. vannamei could be associated with the improved hepatopancreas health associated with M. citrifolia fruit extract supplementation. [ABSTRACT FROM AUTHOR]

Nonylphenol (NP) is one of the common pollutants in the environment that have toxic effects on aquatic animals. Nevertheless, little is known about the possible toxicity mechanism of NP on the hepatopancreas of Litopenaeus vannamei. In the present study, the detrimental effects of NP on the hepatopancreas of the L. vannamei were explored at the histological and transcriptomic levels. The findings indicated that after NP exposed for 3, 12, and 48 h, the hepatopancreas histology was changed significantly. Transcriptomic analysis showed that a total of 4302, 3651, and 4830 differentially expressed genes (DEGs) were identified at 3, 12, and 48 h following NP exposure. All these DEGs were classified into 12 clusters according to the expression patterns at different time points. GO and KEGG enrichment analyses of DEGs were also performed, immunological, metabolic, and inflammatory related pathways, including arachidonic acid metabolism (ko00590), the PPAR signaling pathway (ko03320), and the regulation of TRP channels by inflammatory mediators (ko04750) were significantly enriched. Six DEGs were selected for validation by quantitative real-time PCR (qRT-PCR) and the results confirmed the reliability of transcriptome data. All results indicated that NP is toxic to L. vannamei by damaging the histopathological structure and disrupting the biological function. The findings would provide a theoretical framework for lowering or limiting the detrimental impacts of NP on aquaculture and help us to further study the molecular toxicity of NP in crustaceans.

The giant freshwater prawn (GFP), Macrobrachium rosenbergii, stands out as a vital species in freshwater shrimp aquaculture. However, its susceptibility to cold conditions has increased the cost of aquaculture and limited the growth of GFP cultivation. To furnish insights at the molecular level to enhance GFP's cold tolerance, transcriptome sequencing and physiological analysis of the hepatopancreas were performed in our study. Initially, we identified 4909 differentially expressed genes (DEGs) between experimental (15 °C, 24 h) and control groups (25 °C, 24 h). These DEGs were significantly enriched in the proteasome, spliceosome, lysosome, peroxisome, and lipid metabolism-related pathways including glycerolipid metabolism, sphingolipid metabolism, linoleic acid metabolism as well as steroid hormone biosynthesis. Histological examination following various stress durations revealed a notable increase in hepatocyte vacuoles after 6 h of cold stress. Hepatocyte structure deterioration occurred in the later phase (48 h), coinciding with a significant increase in cell apoptosis. Subsequent analyses demonstrated an initial increase followed by a decrease in superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) content, while malonaldehyde (MDA) content rose with stress duration. Furthermore, a significant increase in triglyceride content and no significant change in cholesterol content was observed. RT-qPCR results unveiled significant down-regulated expression of FAS, TAGL, ACOX1, ACS and significant up-regulated expression of GPAT and DEGS. These findings suggest that oxidative stress and impaired lipid metabolism underlie the poor cold tolerance of GFP.

Abstract Efficient utilisation of plant-based diets in the giant freshwater prawn, Marcrobrachium rosenbergii, varies according to individual, suggesting that it might be associated with differences in physiological and metabolic responses. Therefore, we aimed to investigate the individual differences in the growth response of shrimp fed to a soybean-based diet (SBM). Two hundred shrimp were fed SBM for 90 days, and specific growth rate (SGR) was determined individually. Fast- and slow-growing shrimp (F-shrimp vs. S-shrimp), with the highest and lowest 5% SGRs, respectively, were sampled to determine haemolymph chemistry and carcass composition. The hepatopancreas of these shrimps were used for transcriptome analysis through RNA sequencing (RNA-Seq). The results showed no significant differences in haemolymph chemistry parameters. In terms of carcass proximate composition, F-shrimp exhibited higher protein composition than did S-shrimp, suggesting that F-shrimp have higher protein anabolism. Using RNA-seq and real-time reverse transcription polymerase chain reaction (qRT-PCR), the expression levels of several genes encoding physiologic and metabolic enzymes were found to be upregulated in F-shrimp compared to in S-shrimp, suggesting that these enzymes/proteins mediated the efficient use of SBM-based diets for growth promotion in shrimp. Various DEGs associated with the immune system were observed, indicating a difference in immune processes between F- and S-shrimp. The expression of several housekeeping genes was found to be upregulated in S-shrimp. Collectively, the upregulated expression of several enzymes associated with physiological and/or metabolic processes and increased protein anabolism may be attributed to the efficient use of SBM for maximal growth in shrimp.

The toxic effect and differences of Cr(VI) on superoxide dismutase (SOD) activity, MDA, and metallothionein (MT) content in the hepatopancreas and gill of Portunus trituberculatus were investigated during Cr(VI) enrichment (15 days) and release experiments (15 days). Results showed that the 1.50 and 0.30 mg/L test groups significantly exhibited higher SOD, MDA, and MT content in the hepatopancreas and gill compared with the control group after 15 days of enrichment (p < 0.05). After 15 days of Cr release, the SOD, MDA, and MT content in the hepatopancreas and gill of both test groups recovered to the normal level of the control group (p > 0.05). The gill of P. trituberculatus achieved the highest SOD activity, MDA, and MT content earlier than the hepatopancreas, but the highest values were lower. The gill showed a shorter recovery time than the hepatopancreas. We concluded that the gill of P. trituberculatus exhibited a more rapid response to, and recovery from, Cr(VI) exposure compared to the hepatopancreas, making it a more sensitive tissue for assessing Cr(VI) toxicity, though both tissues showed a capacity for recovery after the removal of the contaminant.

Macrobrachium nipponense, a commercially popular crustacean species within the Chinese context, is recognized for its exceptional nutritional composition and palatability. There are significant differences in growth between male and female M. nipponense. Herein, transcriptomics was used to determine the hepatopancreas transcriptome differences between sex-related size differences in M. nipponense. We identified 974 differentially expressed genes (DEGs) between the SHE (female) and BHE (male) groups, which were validated using RT-qPCR. The genes encoding matrix metalloproteinase-9 (MM9), Ribosome-binding protein 1 (RBP1), Aly/REF export factor 2, and hematological and neurological expressed 1 (HN1) may play a role in modulating the sex-related size differences observed in M. nipponense. Clusters of orthologous groups and gene ontology functional analysis demonstrated that the DEGs for sex-related size in M.nipponense were associated with various biological functions. The Kyoto Encyclopedia of Genes and Genomes pathways analysis demonstrated that upregulated DEGs were mainly enriched in lysine biosynthesis, tryptophan metabolism, and lysine degradation pathways, whereas the downregulated DEGs were mainly enriched in ascorbate and aldarate metabolism, retinol metabolism, and drug metabolism-cytochrome P450 pathways. The results indicated the molecular mechanism underlying the sex-related size differences and identified key genes. This data will be invaluable to support explanations of individual differences between male and female prawns.

Efficient utilisation of plant-based diets in the giant freshwater prawn, Marcrobrachium rosenbergii, varies according to individual, suggesting that it might be associated with differences in physiological and metabolic responses. Therefore, we aimed to investigate the individual differences in the growth response of shrimp fed to a soybean-based diet (SBM). Two hundred shrimp were fed SBM for 90 days, and specific growth rate (SGR) was determined individually. Fast- and slow-growing shrimp (F-shrimp vs. S-shrimp), with the highest and lowest 5% SGRs, respectively, were sampled to determine haemolymph chemistry and carcass composition. The hepatopancreas of these shrimps were used for transcriptome analysis through RNA sequencing (RNA-Seq). The results showed no significant differences in haemolymph chemistry parameters. In terms of carcass proximate composition, F-shrimp exhibited higher protein composition than did S-shrimp, suggesting that F-shrimp have higher protein anabolism. Using RNA-seq and real-time reverse transcription polymerase chain reaction (qRT-PCR), the expression levels of several genes encoding physiologic and metabolic enzymes were found to be upregulated in F-shrimp compared to in S-shrimp, suggesting that these enzymes/proteins mediated the efficient use of SBM-based diets for growth promotion in shrimp. Various DEGs associated with the immune system were observed, indicating a difference in immune processes between F- and S-shrimp. The expression of several housekeeping genes was found to be upregulated in S-shrimp. Collectively, the upregulated expression of several enzymes associated with physiological and/or metabolic processes and increased protein anabolism may be attributed to the efficient use of SBM for maximal growth in shrimp. [ABSTRACT FROM AUTHOR]

During 75 days, the fasting/cyclic refeeding effect was evaluated on survival, growth, and digestive enzymatic activity in the hepatopancreas of freshwater prawn post larvae (PL 20) cultured in a system with/without biofloc. A completely randomized experimental design was used in triplicate to compare: (B3) three-day feeding and three-day fasting; (B6) three-day fasting and six-day refeeding; both cultures in biofloc; (C3) three-day fasting and three-day refeeding; and (C6) three-day fasting and six-day refeeding, both in clear water and two controls with daily feeding were included: (B) culture in biofloc and (C) culture in clear water. The physicochemical variables in water were within the tolerance range for PL's growth. The survival in all treatments was high (>80%) and similar. The most efficient growth was obtained in prawns in B treatments, which was significantly higher than the C treatments, while the growth of the prawns in C treatment was similar to B3 and B6, suggesting that the biofloc covered the nutritional requirements that allowed compensatory growth despite the temporary absence of feed. Higher values of enzymatic activities were observed in the hepatopancreas of prawns in all B treatments, except for acid protease which was similar for all. Prawns in B treatments showed the highest activity of the alkaline protease, in contrast, fewer values were observed in prawns from C3. This study confirmed compensatory growth with the application of biofloc technology in the culture of freshwater prawns in short fasting periods and beneficial effects on growth performance, digestive enzyme activity, and prawn health. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Several studies have reported the high bioindication capacity of Isopoda (Crustacea, Oniscidea), which is related to their important ability to accumulate contaminants, usefulness in soil ecotoxicology and bioindication activities. Any change in the isopod population, diversity and life cycle can indicate relevant pollution levels. The analysis of target tissues, such as the hepatopancreas, is another emerging approach (from a cytologic/histological level) to detect contaminant accumulation from different sources. In this study, tissue disaggregation procedures were optimised in the hepatopancreas, and flow cytometry (FC) was applied to detect cell viability and several cell functions. After disaggregation, two hepatopancreatic cell types, small (S) and big (B), were still recognisable: they differed in morphology and behaviour. The analyses were conducted for the first time on isopods from sites under different conditions of ecological disturbance through cytometric re-interpretation of ecological-environmental parameters. Significant differences in cell functional parameters were found, highlighting that isopod hepatopancreatic cells can be efficiently analysed by FC and represent standardisable, early biological indicators, tracing environmental-induced stress through cytologic/histologic analyses. [ABSTRACT FROM AUTHOR]

Vitellogenin (Vtg) is a precursor of yolk proteins in egg-laying vertebrates and invertebrates and plays an important role in vitellogenesis and embryonic development. However, the Vtg family remains poorly characterized in Exopalaemon carinicauda, a major commercial mariculture species found along the coasts of the Yellow and Bohai Seas. In this study, 10 Vtg genes from the genomes of E. carinicauda were identified and characterized. Phylogenetic analyses showed that the Vtg genes in crustaceans could be classified into four groups: Astacidea, Brachyra, Penaeidae, and Palaemonidae. EcVtg genes were unevenly distributed on the chromosomes of E. carinicauda, and a molecular evolutionary analysis showed that the EcVtg genes were primarily constrained by purifying selection during evolution. All putative EcVtg proteins were characterized by the presence of three conserved functional domains: a lipoprotein N-terminal domain (LPD_N), a domain of unknown function (DUF1943), and a von Willebrand factor type D domain (vWD). All EcVtg genes exhibited higher expression in the female hepatopancreas than in other tissues, and EcVtg gene expression during ovarian development suggested that the hepatopancreas is the main synthesis site in E. carinicauda. EcVtg1a, EcVtg2, and EcVtg3 play major roles in exogenous vitellogenesis, and EcVtg3 also plays a major role in endogenous vitellogenesis. Bilateral ablation of the eyestalk significantly upregulates EcVtg mRNA expression in the female hepatopancreas, indicating that the X-organ/sinus gland complex plays an important role in ovarian development, mostly by inducing Vtg synthesis. These results could improve our understanding of the function of multiple Vtg genes in crustaceans and aid future studies on the function of EcVtg genes during ovarian development in E. carinicauda. [ABSTRACT FROM AUTHOR]

The microstructure of the stomach, intestines, hepatopancreas, kidneys, lungs, reproductive organs, and protein gland of two species of snails (Helix pomatia and Helix aspersa Muller) under the influence of abiotic (climatic) and biotic (parasite) factors was studied. In snails of the first group, the visceral organs had a typical structure. The parenchyma of the hepatopancreas, as the most vulnerable organ, is represented by glandular tubules and the system of excretory ducts that open into the intestinal cavity. Structural changes in the hepatopancreas and gonads were observed in snails of the second group, which were in the state of anabiosis under the influence of dry, hot weather. The helminthic invasion in the snails of the third experimental group caused necrosis of the hepatopancreas and replacement of the parenchyma of the gland with loose connective tissue. [ABSTRACT FROM AUTHOR]

Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most used pharmaceuticals in the world that introduced into the environment via hospital wastes. These drugs have been detected in many aquatic ecosystems in Egypt, such as the Rosetta Branch of the River Nile, even in drinking water. The objective of this study is to investigate the toxicological effects of ibuprofen (IBP), diclofenac (DCF), and naproxen (NPX) on the freshwater crayfish Procambarus clarkii at concentrations that are close to those in its natural environment. The experimental crayfish were exposed for 60 days and divided into 9 groups; including the control, DCF, IBP, and NPX separately or in combination with the three drugs. The used low concentrations were 0.64mg/L, 100µg/L, and 2mg/L, while high ones were 17mg/L, 63mg/L, and 15mg/L for DCF, IBP, and NPX, respectively. The eighth group of crayfish was treated with a mixer of drugs at low doses while the ninth was treated with a previous mixer co-treated with Chlorella vulgaris algae. The results showed that lipid peroxidation, nitric oxide contents, glutathione-S-transferase, aspartate transaminase, alanine transaminase, and alkaline phosphatase activities were significantly increased in the hepatopancreas of the crayfish exposed to IBP, DCF, and NPX. On the other hand, total protein, glucose, and calcium contents were significantly decreased after IBP, DCF, and NPX exposure versus the control. Biological treatment with C. vulgaris improved marginally these symptoms. Also, obvious histological damage was observed by drug treatments and slight improvement was noticed by adding algae. According to these findings, DCF, IBP, and NPX were to blame for the abnormalities in the biochemical indicators examined and the histological damage at low and high doses. While treatment with C. vulgaris adjusted the damage in the mixed low dose although this improvement was not to the level of the control. [ABSTRACT FROM AUTHOR]

The hepatopancreas of Procambarus clarkii provides lipids and other nutrients for ovarian development and contains hormones such as estradiol related to ovarian development. During the development of the P. clarkii ovary, the hepatopancreas' tissue structure and fatty acid content undergo corresponding changes. A controlled indoor experiment was conducted to investigate the effects of exogenous estradiol injection on the tissue structure and fatty acid content in the hepatopancreas of P. clarkii. The experiment consists of three experimental groups and one control group. The P. clarkii in the experimental group were injected with 5, 0.5, and 0.05 µg/(g body weight) of estradiol once every 5 days for 5 times and with normal saline injection as the control. After 25 days, the content of fatty acids and changes in tissue structure in the hepatopancreas were measured. The results showed that the main fatty acids in P. clarkii hepatopancreas were C18:4n, C20:3n, and EPA, and polyunsaturated fatty acid (PUFA) content was 57.17%±4.76%. Injecting estradiol mainly affected the content of C16:0, C16:1n, C18:0, C18:1n9, C18:2n, C18:3n6, C18:3n6, C18:4n, C20:1n, C20:2n, C20:3n, C20:4n3, C20:5n3, C22:n (n=1-4), C22:5n, and C22:6n in the hepatopancreas. Injecting estradiol significantly reduced the total content of PUFA in hepatopancreas with the reduction of C18:3n6, C18:4n, C20:3n, C20:4n3, C20:5n3 C22: n (n=1-4) (P<0.05); Estradiol injection significantly increased the total content of monounsaturated fatty acid (MUFA) in hepatopancreas with the increase of C18:1n9, and C20:1n (P<0.05); Injection of estradiol (0.05 µg/g) significantly increased the total content of saturated fatty acids (SFA) in the hepatopancreas (P<0.05) with the increase of C16:0 and C18:0 (P<0.05). Injecting estradiol can increase the volume and quantity of hepatopancreatic B cells, and the 5 µg/g group had the most obvious changes. [ABSTRACT FROM AUTHOR]

The oriental river prawn Macrobrachium nipponense is an important freshwater economic species in China, producing huge economic benefits. However, M. nipponense shows lower alkali tolerance than fish species, thus genetic selection is urgently needed in order to improve alkali tolerance in this species. In the present study, the effects of alkalinity exposure on the hepatopancreas of M. nipponense were measured under the alkali concentrations of 0 (control), 4, 8, and 12 mmol/L with the exposure time of 96 h through histological observations, measurement of antioxidant enzymes, metabolic profiling analysis, and transcriptome profiling analysis. The present study identified that the low concentration of alkali treatment (<4 mmol/L) did not result in morphological changes in the hepatopancreas and activity changes in antioxidant enzymes, while high-alkali treatment (>8 mmol/L) damaged the normal structures of the lumen and vacuoles and significantly stimulated the levels of superoxide dismutase, catalase, and total antioxidant capacity, indicating these antioxidant enzymes play essential roles in the protection of the body from the damage caused by the alkali treatment. Metabolic profiling analysis revealed that the main enriched metabolic pathways of differentially expressed metabolites in the present study were consistent with the metabolic pathways caused by environmental stress in plants and other aquatic animals. Transcriptome profiling analysis revealed that the alkali concentration of <8 mmol/L did not lead to significant changes in gene expression. The main enriched metabolic pathways were selected from the comparison between 0 mmol/L vs. 12 mmol/L, and some significantly up-regulated genes were selected from these metabolic pathways, predicting these selected metabolic pathways and genes are involved in the adaptation to alkali treatment in M. nipponense. The expressions of Ras-like GTP-binding protein, Doublesex and mab-3 related transcription factor 1a, and Hypothetical protein JAY84 are sensitive to changes in alkali concentrations, suggesting these three genes participated in the process of alkali adaptation in M. nipponense. The present study identified the effects of alkalinity exposure on the hepatopancreas of M. nipponense, including the changes in antioxidant status and the expressions of metabolites and genes, contributing to further studies of alkali tolerance in this species. [ABSTRACT FROM AUTHOR]

Background: The rock goby, Gobius paganellus, is not a commercial species. This species has an essential role in the coastal ecosystem as a biological indicator. Therefore, it has been selected as the study's model species. Aim: Due to the insufficient studies that have described the hepatopancreas of G. paganellus, this study aimed to provide information on the anatomical and histological structure of the hepatopancreas of the alimentary canal of this species on the western coast of Libya. Methods: Fifty mature G. paganellus specimens were collected from the northwest of Libya (Tajoura, Jodaem, and Farwa Island). Total length and total weight of the samples were measured and performed by using gross anatomy and histology. Then, the histological sections (3-5 µm) were stained with hematoxylin and eosin (H and E). Results: Morphologically, the liver has a large pyriform lobe. The dorsal surface of the liver is shiny and smooth, and the ventral surface contains shallow sulci; each sulus has a large blood vessel. Histologically, the liver is wrapped with a thin capsule of fibro-connective tissue. The hepatic parenchyma is made of hepatocytes with blood sinusoids. The hepatocytes are polygonal-shaped cells and have no hepatic lobules or portal triads. Melano-macrophage centers are distributed next to the blood vessels and bile ducts. The bile ducts are lined by columnar epithelial cells. The exocrine pancreatic tissue was observed in the liver parenchyma, and it consists of acini that are composed of pyramidal cells and contain zymogen granules. Conclusion: The liver of this species has both pancreatic and liver tissue, which was discovered in this investigation for the first time. [ABSTRACT FROM AUTHOR]

The supplementation of cytosine phosphate guanosine-oligodeoxynucleotides (CpG-ODNs) in feeds to enhance the immune responses of crustaceans has been studied. However, the immunomodulatory effects of CpG-ODNs alone on prawn hepatopancreas and midgut remain unclear. Hence, we delivered CpG-ODNs directly into the midgut of Macrobrachium rosenbergii by reverse gavage (RG). In this study, M. rosenbergii was fasted before receiving different doses (0, 2, 4, and 10 μg) of CpG-ODN 2006. Furthermore, the hemolymph, hepatopancreas, and midgut were sampled at 2, 6, 12, and 24 h after RG. The results showed that administration of CpG-ODN 2006 could boost total hemocyte count (THC), transglutaminase (TG), phenoloxidase (PO), lysozyme-like (LYS), and respiratory burst (RB) responses in the hemolymph. In addition, the expression levels of prophenoloxidase (MrProPO), crustin (MrCrs), peroxinectin (MrPxt), and superoxide dismutase (MrSOD) in the hepatopancreas and midgut were also enhanced, in a dose- and time-dependent manner. Overall, our results indicate that administration of 4 μg CpG-ODN 2006 alone by RG elicited the most potent immunological response in the hemolymph, hepatopancreas, and midgut of M. rosenbergii. [ABSTRACT FROM AUTHOR]

The study focuses on the isolation, characterization, and expression analysis of a lectin from the hepatopancreas of Macrobrachium rosenbergii. The protein was isolated by affinity chromatography on a melibiose-agarose column. The molecular weight of the native protein was found to be ~120 kDa which consists of a single polypeptide of ~39.5 kDa. On mass spectrometric analysis, the protein was identified as lipopolysaccharide- and beta-1,3-glucan binding protein (LGBP). LGBP showed hemagglutination with rabbit RBC like a lectin and its carbohydrate-binding specificity was determined by the hemagglutination inhibition test. The protein also showed antibacterial activity against two Gram-negative bacteria Vibrio harveyi and Aeromonas sobria, and one Gram positive bacteria Bacillus cereus in the disc diffusion test. Rabbit antiserum was raised against the purified LGBP and used to develop a sandwich ELISA system for quantitation of the protein in hepatopancreas and serum samples of M. rosenbergii. The expression of the LGBP transcripts in muscle, hepatopancreas, and gill tissues from M. rosenbergii juveniles at 72 h post-challenge of V. harveyi was not modulated as noticed in qPCR analysis. However, significant increases in the concentrations of LGBP protein in hepatopancreas (5.23 ± 0.45 against 3.43 ± 0.43 mg/g tissue in control) and serum (1.08 ± 0.14 against 0.61 ± 0.08 µg/ml in control) were observed in the challenged group of prawns in ELISA suggesting its putative role against bacterial infections. The study for the first time characterized the native LGBP of M. rosenbergii showing a multifunctional role in immunity. [ABSTRACT FROM AUTHOR]

Human enteric viruses, such as hepatitis A virus (HAV), hepatitis E virus (HEV), and norovirus genogroups I and II (NoVGI and NoVGII), cause infections, and it has been largely demonstrated that mussels play an important role if consumed as raw or undercooked food matrices. This study aimed to investigate, through qualitative and quantitative biomolecular assays, the detection of partial genomic regions belonging to the most relevant enteropathogenic viruses for humans (HAV, HEV, NoVGI and NoVGII) in mussels (Mytilus galloprovincialis) farmed along the coasts of two Italian regions on the central Adriatic Sea: Abruzzo (Casalbordino, Chieti) and Molise (Termoli, Campobasso). A total of 425 animals were sampled, and the respective georeferentiations were registered. A total of 85 pools, each composed of five subjects/aliquot, were formed (22 from Abruzzo and 63 from Molise regions). This step was followed by homogenization and RNA extraction, and then the biomolecular assays [nested reverse transcription polymerase chain reaction (PCR) and real-time reverse transcription-quantitative PCR] were performed. 1.17% of the pool was positive for HAV RNA detection (102 copies/mL), 9.41% for HEV (102-103 copies/mL), 2.35% for NoVGI (101 copies/mL), and no pool was positive for NoVGII. This study demonstrated the human enteric viruses’ presence in mussels farmed in a low-investigated marine area. Based on a one-health point of view, this paper wants to enforce the importance of biomolecular and epidemiological screenings as surveillance systems to guarantee human, animal, and environmental health.

Lipid accumulation in ovaries is essential for reproductive success in decapods. Understanding of the mechanism of ovarian lipid accumulation is still in its infancy. In this study, we investigated lipid compositions of ovaries and hepatopancreas throughout vitellogenesis and hemolymph in the late-vitellogenic stage in the mud crab Scylla paramamosain. Total lipid content in ovaries consecutively increased from the early to late stage, and the main accumulated lipids were phosphatidylcholine (PC), alkenylglycerophosphoethanolamine (alkenyl-PE), triacylglycerol (TG), cholesteryl ester, and cholesterol. The lipid composition of hemolymph indicated that PC, alkenyl-PE, diacylglycerol (DG), ether DG, monoacylglycerol, and cholesterol were the main lipids transferred to ovaries from hepatopancreas. In hepatopancreas, the total lipid content appeared to increase during vitellogenesis and the late stage was significantly different from early and mid-stages. TG was the predominant lipid class that slightly decreased from the early to mid-stage, but dramatically increased in the late stage. In contrast, PC and alkenyl-PE significantly increased from the early to mid-stage and remained constantly in relatively low proportion until the late stage. These findings suggested PC and alkenyl-PE in the hepatopancreas likely play critical roles in regulating lipid transfer. Esterified fatty acids in ovaries and hepatopancreas were mainly C16:0, C18:0, 16:1, C18:1, C18:2, C20:4, C20:5, and C22:6, of which highly unsaturated fatty acids (HUFA) primarily occurred in alkenyl-PE and phosphatidylserine (PS) but not PC. In hemolymph, PC was abundant in HUFA besides of saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA), while PS was predominantly composed of SFA and MUFA, implying a transformation of HUFA between phospholipids prior to transfer. These findings highlight the importance of phospholipid metabolism in the hepatopancreas to ovarian lipid accumulation. Our study provides novel insights into the mechanism of ovarian lipid accumulation in decapods.

The crustacean hepatopancreas is a complex organ, with multiple functions that are essential for coping with adverse environments. Excessive alkalinity in water seriously threatens the survival of crustaceans, however, studies on its mechanism of alkalinity stress and adaption are still limited. In this study, we aimed to investigate the mechanism of acute alkalinity stress in hepatopancreas of Pacific white shrimp Litopenaeus vannamei (initial weight: 0.22±0.13 g) by integrating physiological, histological, transcriptome, and metabolome analyses. Groups performed acute alkalinity stress for 24 h, including control (alkalinity: 45 mg/L) and treatment (alkalinity: 350 mg/L). The physiological parameters related to energy metabolism, ammonia detoxification, and oxidative stress were significantly changed under acute alkalinity stress, including glycogen, glucose, fatty acid synthesis, glucose dehydrogenase, glutamine synthase, urea, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde. The expression levels of genes and metabolites were significantly changed, which related to glycolysis/gluconeogenesis, fatty acid metabolism, glycerophospholipid metabolism, amino acid metabolism, ammonia metabolism, and antioxidation. In addition, the local structure of the hepatopancreas was damaged and accompanied by cell apoptosis. The above results indicate that the glycogen catabolism in hepatopancreas was significantly enhanced under acute alkalinity stress, while the lipid anabolism was inhibited. Additionally, the ornithine-urea cycle mechanism was further mobilized to alleviate the ammonia excretion and accumulation in the internal environment. The antioxidant capacity and glycerophospholipid metabolism were disturbed in hepatopancreas. In addition, acute alkalinity stress would induce apoptosis in hepatopancreas. The current study will provide important clues for understanding the stress response mechanism of Pacific white shrimp under acute alkalinity stress and lay a theoretical basis for the optimization of related saline-alkaline aquaculture.

Background: The rock goby, Gobius paganellus is not a commercial species. This species has an essential rule in the coastal ecosystem as a biological indicator. Therefore, it has been selected as study's model species. Aim: Due to the insufficient studies that have been described on the hepatopancreas of G. paganellus, this study aimed to provide information on the anatomical and histological structure of the hepatopancreas of the alimentary canal of this species in the western coast of Libya. Methods: Fifty mature G. paganellus specimens were collected from the northwest of Libya (Tajoura, Jodaem, and Farwa Island). Total length and total weight of the samples were measured, and performed by using gross anatomy, and histology. Then, the histological sections (3-5 µm) were stained with hematoxylin and eosin (H and E). Results: Morphologically, the liver has a large pyriform lobe. The dorsal surface of the liver is shiny and smooth, and the ventral surface contains shallow sulci; each sulus has a large blood vessel. Histologically, the liver is wrapped with a thin capsule of fibro-connective tissue. The hepatic parenchyma is made of hepatocytes with blood sinusoids. The hepatocytes are polygonal-shaped cells and have no hepatic lobules or portal triads. Melano-macrophage centers are distributed next to the blood vessels and bile ducts. The bile ducts are lined by columnar epithelial cells. The exocrine pancreatic tissue was observed in the liver parenchyma and it consists of acini that are composed of pyramidal cells and contain zymogen granules. Conclusion: The liver of this species has both pancreatic and liver tissue, which was discovered in this investigation for the first time. [Open Vet J 2023; 13(10.000): 1251-1258]

Abstract Background The hepatopancreas of crustaceans serves as a significant organ for both the synthesis and secretion of digestive enzymes, as well as energy storage. In the event of food shortage, the hepatopancreas can provide energy for survival. To investigate the potential regulatory mechanisms of the hepatopancreas in response to starvation in Eriocheir Sinensis, transcriptome analysis, histological study and qRT-PCR were performed. Results The results showed that starvation caused a decrease in the hepatopancreas index of E. sinensis, which had certain effects on the tissue structure, metabolism and angiogenesis in the hepatopancreas. In addition, WGCNA and linear regression analysis showed that the genes significantly related to the hepatopancreas index were mainly enriched in the angiogenesis pathway, in which AKT signaling played an important role. Starvation may inhibit AKT signaling pathway by reducing the expression of TGFBI, HSP27, HHEX, and EsPVF1, thereby hindering angiogenesis, promoting apoptosis, and leading to hepatopancreas atrophy. Conclusion These results indicate that AKT plays an important role in the angiogenesis pathway and apoptosis of the starvation induced hepatopancreas index reduction, which is beneficial to further understand the effect of starvation stress on hepatopancreas of Chinese mitten crab.

ABSTRACT Large quantities of heavy metals come from anthropogenic sources, including coal combustion, furthermore, these heavy metals can cause oxidative stress in terrestrial animals, such as snails, which acquire these metals through food from storage plants in industrial and coal-burning-polluted areas. In this research project, we measured the impact and distribution of power plant TC Kosova A's activity on the concentrations of heavy metals (Pb, Zn, Ni) in the locality of Obiliq, as well as their effect on oxidative stress parameters such as carbonyl proteins, malondialdehyde (MDA), and total proteins in the hepatopancreas of the snail Helix pomatia L. We collected 120 soil samples and 12 snails from concentric circles around the pollution point at distances of 1 km, 2 km, and 5 km. Relevant methods were then applied, and the samples were measured using a spectrophotometer and flame absorber Analyticyena. Our results shown that the heavy metals Pb, Zn, and Ni bioaccumulated from snail shells (Helix pomatia L.), and concentrations of these metals influenced an increase in the levels of oxidative stress parameters, including protein carbonylation, malondialdehyde (MDA), and total proteins in the hepatopancreas.

Transport is an essential part of the aquaculture and research of the main freshwater aquaculture crayfish Procambarus clarkii in China. However, transport is often accompanied by a low survival rate. Assessing the physiological state of P. clarkii before and after transport may discover the cause of this high mortality rate. In this study, ice-cold and exposed-to-air transport methods were compared using an array of parameters, including relative expression level of heat shock protein 70 (HSP70), content of serum glucose and cortisol, immune parameters (enzyme and immune-related genes), and white spot syndrome virus (WSSV) concentration were investigated to understand the physiological state of P. clarkii before and after transport, as well as the cause of dying crayfish on days 5 and 7 after transport stress. Histological sections of hepatopancreas, gills, and intestines reflected pathological changes. The survival rate of crayfish with ice-cold transport was significantly higher than that with exposed-to-air transport, and mortality peaked at 3–9 days after transport stress. A prolonged response to oxidative stress and short-term immunosuppression was present after transport, and the trend of the WSSV concentration in the hepatopancreas was similar to the mortality rate of P. clarkii. The contents of serum glucose and cortisol, antioxidant enzymes and immune-related indexes, and the concentration of WSSV in hepatopancreas of dying crayfish were significantly higher than those of vibrant crayfish on the 5th and 7th days after transport. The hepatopancreas, intestines, and gills of dying crayfish had varying degrees of damage, and the hepatopancreas and intestines were severely damaged. The results suggested that the death of P. clarkii after transport stress is caused by oxidative stress, the imbalance of reactive oxygen species regulation, and decreased WSSV resistance, which eventually led to irreversible tissue damage. The increase of WSSV in the body of crayfish might be the direct cause of crayfish death.

A high-performance liquid chromatography method coupled to tandem mass spectrometry was validated in order to study the pharmacokinetics of cefotaxime in shrimp hepatopancreases and plasma, as well as its withdrawal time related to a maximum residue limit (MRL) in shrimp muscle. Pharmacokinetics parameters were investigated through oral medication at a single dose of 25 mg/kg shrimp body weight and subsequent hepatopancreas and plasma cefotaxime concentration measurements at 0.5, 1, 2, 4, 8, 12 and 24 h after shrimp were fed with medication. The maximum concentration of cefotaxime was observed after one hour in the hepatopancreas (Cmax, 19.45 ± 2.10 mg/kg) and 4 h in plasma (0.184 ± 0.061 mg/L). Based on a minimum inhibitory concentration (MIC) of cefotaxime of 4.13 mg/L against Vibrio parahaemolyticus (known to cause acute hepatopancreatic necrosis disease (AHPND) in white leg shrimp), it was observed that the time during which the hepatopancreas cefotaxime concentration was above the MIC was 23 h. An every 24 h cefotaxime treatment could thus be effective in fighting against this bacterium in shrimp. The withdrawal time of cefotaxime was determined after shrimp were fed with medicated feed once a day and twice a day for three consecutive days. Shrimp muscle was collected on day 1 and day 3 during medication and 1, 3, 7, 14 and 21 days after medication was stopped. Considering an MRL of 50 μg/kg, the withdrawal times were 8.5 degree-days (corresponding to 6.9 h at 29.5 °C) after shrimp were fed with medicated feed once a day for 3 days and 95.5 degree-days (77.7 h at 29.5 °C) after shrimp were fed with medicated feed twice a day for 3 days. Moreover, histological analysis revealed that feeding shrimp with cefotaxime at the given dose in once- or twice-a-day treatments did not negatively impact the shrimp hepatopancreas.

ObjectiveTo understand the concentration of heavy metal cadmium and cadmium in portunus and mantis shrimp， and to timely identify food safety problems and potential hazards.MethodsPortunus and mantis shrimp samples from different provinces were collected and categorized based on different regions and locations， and some samples were made from tissue parts. Graphite furnace atomic absorption spectrometry was used to detect cadmium items， the cadmium exposure of portunus and mantis shrimp was evaluated simultaneouslyResultsThe detection rate of cadmium in 124 batches of portunus sold in Shanghai was 100% （124/124）， the detection rate of cadmium in 63 batches of mantis shrimp sold in the market was also 100% （63/63）. The cadmium content varied in different tissue parts， and the cadmium enrichment in hepatopancreas was the highest in the edible parts of portunus and mantis shrimp. The average detection value， 50th percentile value， 95th percentile value of cadmium in the hepatopancreas of portunus accounted for 52.64%， 49.28% and 98.65% of the PTMI， respectively. The average detection value， 50th percentile value and 95th percentile value of cadmium in the hepatopancreas of mantis shrimp accounted for 30.76%， 32.04% and 46.16% of the PTMI， respectively.ConclusionThe average residual levels of heavy metal cadmium in portunus and mantis shrimp are within the safe range.

Chinese herbs and their purified components in aquafeeds have been proved effective to combat stress in aquatic animals to combat toxic environmental stress. Lycium barbarum poly-saccharides (LBP) is an active antioxidant, anti-inflammatory and antitoxic purified herbal agent. The objective of this paper was to evaluate the effect of 0.1–2.0% LBP on the hepatopancreas of Chinese mitten crab (Eriocheir sinensis) challenged with 2 mg/kg T-2 toxin orally for 5 weeks. The results showed that feeding LBP significantly restored the hepatopancreas index (HI), gonadsomatic index (GSI) reduced by T-2 toxin. Moreover, H&E sections indicated that LBP dose-dependently restored the disrupted microstructure of the hepatopancreas. In antioxidative capacity, significant increases in malondialdehyde (MDA) reduction and elevated glutathione (GSH) content, gluta-thione reductase (GPx) activity were seen in the 0.5% LBP group. In addition, reactive oxygen species (ROS) and apoptosis rate of hepatopancreas showed the anti-apoptotic effect of LBP doses, plus qPCR assay of 10 apoptosis-related genes to support them. The mRNA abundance of three pro-inflammatory genes was significantly reduced by LBP, corroborating its anti-inflammatory efficacy. In brief, feed supplementation with LBP alleviated the oxidative stress-apoptosis-inflammation induced by co-feeding T-2 toxin in E. sinensis, maintained the structural and functional integrity of the hepatopancreas, prevented degenerative atrophy.

The effect of moderate hypoxia on the processes of energy metabolism was studied in the tissues (gills, hepatopancreas) of the Mediterranean mussel Mytilus galloprovincialis (Lamarck, 1819). The mollusks was kept at 6.8–6.9 mg O2 L–1 (control group) or 1.9–2.0 mg O2 L–1 (experimental group). In both cases, water temperature was 22 ± 1°C, salinity 17–18‰, exposure time 72 h. Dissolved oxygen content in water was lowered by nitrogen bubbling for 4–5 h. Under moderately hypoxic conditions, a complex of biochemical reactions aimed at retaining the baseline energy status developed in the mussel tissues. While aerobic processes were clearly on the decline, as evidenced by a decrease in succinate dehydrogenase (SDH) activity, anaerobic glycolysis intensified. Aldolase and malate dehydrogenase (MDH) activities, as well as tissue pyruvate level, increased. Both lactate dehydrogenase (LDH) activity and lactate levels in mussel tissues remained unchanged. The state of the M. galloprovincialis adenylate system was comprehensively characterized. The tissue content of the ATP fraction remained at the level of control values, reflecting an adaptive trend in the reorganization of tissue metabolism. The ability of the hepatopancreas to accumulate ADP and AMP fractions from the circulatory systems under conditions of experimental hypoxic load was noted for the first time. Presumably, this may largely explain the increase in the pool of adenylates and the decrease in the adenylate energy charge in this organ. It was concluded that the reorganization of carbohydrate metabolism in M. galloprovincialis under moderately hypoxic conditions allows the mussel to retain the energy status of its tissues and, therefore, to populate challenging (hypoxic) waters. [ABSTRACT FROM AUTHOR]

This is an open access article under the CC BY-NC-SA license Enterocytozoon hepatopenaei (EHP) is a spore-forming microsporidia intracellular parasite, which slough off the cells of the hepatopancreases of the shrimp. This study was intended to determine the effect of immersion duration and maintenance time to stimulate the immune system of shrimps by using a crude protein from Zoothamnium penaei to protect against EHP infections. The study used a completely randomized design where shrimps were divided into four groups each having five replicates. Groups P1, P2, P3 were immersed in immunostimulant for 10, 15, and 20 minutes respectively, except negative control. The phenoloxidase, phagocytosis activities, histopathology study were determined. Also, the molecular diagnosis of EHP was measured to confirm EHP infection in the shrimp samples. The phenoloxidase activity significantly increased after immersion for 20 minutes (P3) from 0.64-, 0.72-, and 0.78-units min-1 mil-1 of protein on the first week, second week and third week, respectively. The phagocytosis activities of the sample immersed for 20 minutes were significantly increased to 68% on the second week. The sample also showed improved hepatopancreases epithelial cells with only 26% necrotic cell, less EHP spore and vacuolation. There was an interaction between immersion duration and maintenance time of the L. vannamei immersed in crude protein from Zoothamnium penaei. Twenty minutes immersion were significantly found to stimulate the immune system of the shrimp against EHP. The present work revealed that, the application of crude protein from Z. penaei is effective against EHP in shrimp culture. [ABSTRACT FROM AUTHOR]

Crustaceans have successfully adapted to survive in their natural habitat, rich in microorganisms, due to the presence of antimicrobial peptides (AMPs) in their organism. They achieve this adaptation despite lacking the highly specific adaptive immune system found in vertebrates. One valuable source of AMPs is the hepatopancreas, a waste product from crab fishery and its processing. Applying zymographic and spectrophotometric techniques, we discovered a small peptide (approximately 5 kDa) within a low molecular weight protein fraction extracted from the acetone powder of the red king crab hepatopancreas. This peptide hydrolyzes both M. lysodeikticus cell wall and M. lysodeikticus cell wall polysaccharide, while showing no activity against gelatin. The found peptide may be of interest for application in medicine, biotechnology, and the food industry, for example as a bio-preservative. [ABSTRACT FROM AUTHOR]

Background: The hepatopancreas of crustaceans serves as a significant organ for both the synthesis and secretion of digestive enzymes, as well as energy storage. In the event of food shortage, the hepatopancreas can provide energy for survival. To investigate the potential regulatory mechanisms of the hepatopancreas in response to starvation in Eriocheir Sinensis, transcriptome analysis, histological study and qRT-PCR were performed. Results: The results showed that starvation caused a decrease in the hepatopancreas index of E. sinensis, which had certain effects on the tissue structure, metabolism and angiogenesis in the hepatopancreas. In addition, WGCNA and linear regression analysis showed that the genes significantly related to the hepatopancreas index were mainly enriched in the angiogenesis pathway, in which AKT signaling played an important role. Starvation may inhibit AKT signaling pathway by reducing the expression of TGFBI, HSP27, HHEX, and EsPVF1, thereby hindering angiogenesis, promoting apoptosis, and leading to hepatopancreas atrophy. Conclusion: These results indicate that AKT plays an important role in the angiogenesis pathway and apoptosis of the starvation induced hepatopancreas index reduction, which is beneficial to further understand the effect of starvation stress on hepatopancreas of Chinese mitten crab. [ABSTRACT FROM AUTHOR]

This study aimed to explore the effects of different starch source diets on the growth performance and hepatopancreas RNA-seq of grass carp. Juvenile grass carp (initial body weight of 39.4 ± 1.6 g) were fed diets containing 25% corn (CO), potato (PO), and wheat (WH) starch for 8 weeks, respectively. The weight gain ratio (WGR) was significantly lower, whereas the visceral somatic index (VSI) and feed conversion ratio (FCR) were significantly higher in the CO group than those in the PO and WH groups. These indicators did not significantly differ between the PO and WH groups. Hepatopancreas RNA-seq analysis showed that 536, 514, and 647 differentially expressed genes (DEGs) were screened out in the comparisons of PO vs. WH, PO vs. CO, and CO vs. WH. The DEGs were mainly enriched in the several known pathways involved in steroid biosynthesis, cell cycle, fatty acid metabolism, and fat digestion and absorption according to Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. The major DEGs related to lipid and carbohydrate metabolism were analyzed, in which lipogenesis-related DEGs (fasn, acc1, scd1, elovl6, and me1), fat digestion and absorption-related DEGs (fabp7, apoa1, apoa4, and pla2), and glycometabolism-related DEGs (gk, g6pd, and pepck) were down-regulated in the PO group compared with those in the CO and WH groups. Conversely, steroid synthesis-related DEGs (hmgcs, fdft1, sqle, lss, cyp51, msmo1, nsdhl, ugt, cyp1b1, and cyp7a1) were up-regulated in the PO group. These results indicate that the long-term PO ingestion could modulate hepatic lipid metabolism by reducing fatty acid biosynthesis and increasing bile acid biosynthesis. PO may be healthier in contrast to CO alone, which may not be suitable as a starch source in grass carp diet. [ABSTRACT FROM AUTHOR]

The aim: study the effect of the synthetic anabolic steroid 19-nortestosterone on the fractional composition of blood plasma proteins, as well as the total protein and albumin content, indicators of the metabolism of carbohydrates, lipids, macro- and microelements in the fish hepatopancreas. Materials and methods. The experiments were conducted on two-year-old carp (Cyprinus carpio L.), which were kept for 24 hours in aquariums with a volume of 40 dm³ with different concentrations of 19-nortestosterone in water at optimal temperature, oxygen content, and pH. Spectrophotometric methods and an automatic biochemical analyzer were used to determine metabolic indicators in fish, and disk electrophoresis in PAGE was used to determine the fractional composition of proteins. Results. It was established that the steroid hormone 19-nortestosterone, the concentration of which in the water of the fish of the experimental groups was 50 and 200 µg/dm³, respectively, increased the level of total protein and albumin, the concentration of glucose, triglycerides, creatinine, inorganic phosphorus and iron in the hepatopancreas and did not affect the calcium content. In the blood plasma of carp, kept in water with a concentration of 19-nortestosterone of 50 µg/dm³, the content of proteins with a molecular weight of 240-231 increases; 215-179; 169-146 and 105-95 kDa, decreases - 35-33 kDa, fractions of proteins with a molecular weight of 228-220 appear; 116-105 and 48-44 kDa. An increase in the concentration of 19-nortestosterone in water to 200 µg/dm³ increased the content of proteins with a molecular weight of 240-231 and 215-179 kDa in the blood plasma of fish, and decreased it by 35-33 kDa, causing the redistribution of individual protein fractions due to the appearance of proteins with a molecular weight of 240-228; 116-105 and 48-44 kDa in the absence of proteins 42-41; 38-36 and 24-20 kDa. Conclusions. Based on the obtained results, it was concluded that the anabolic synthetic steroid 19- nortestosterone stimulates the biosynthesis of proteins, the metabolism of carbohydrates and lipids, and its effect on the content of minerals in the hepatopancreas and the fractional composition of blood plasma proteins in carp fish was proven [ABSTRACT FROM AUTHOR]

Recent research on decapod crustaceans revealed the presence of multiple vitellogenin genes within a single species. Characterizing their expression dynamics and regulatory mechanisms is helpful for the sustainability of commercially relevant decapods through a greater comprehension of vitellogenesis. However, it is unclear whether multiple vitellogenin genes exist in the kuruma prawn Marsupenaeus japonicus, an essential species in several Asian countries. This study determined the full-length cDNA sequence of the new vitellogenin (Maj-Vg2) using transcriptome analysis and cDNA cloning. The deduced Maj-Vg2 consisted of 2,554 amino acids and shared only 53.8% identity with Maj-Vg1, the known vitellogenin of this species. The Maj-Vg1 gene (Maj-Vg1) was expressed in both the hepatopancreas and ovary, whereas the Maj-Vg2 gene (Maj-Vg2) was expressed only in the hepatopancreas. Maj-Vg2 expression in the hepatopancreas was low during the previtellogenic stage and increased during the early and late yolk globule stages. When eyestalks were ablated from immature prawns, Maj-Vg2 expression in the hepatopancreas increased significantly, like Maj-Vg1 expression in the ovary, suggesting possible regulation by hormones from the eyestalk ganglia. These results suggested that Maj-Vg2 and Maj-Vg1 are involved in vitellogenesis in this species. [ABSTRACT FROM AUTHOR]