Crystal structure of the iron-sulfur cluster transfer protein ApbC from Escherichia coli.

Iron-sulfur (Fe-S) clusters are ubiquitous and are necessary to sustain basic life processes. The intracellular Fe-S clusters do not form spontaneously and many proteins are required for their biosynthesis and delivery. The bacterial P-loop NTPase family protein ApbC participates in Fe-S cluster assembly and transfers the cluster into apoproteins, with the Walker A motif and CxxC motif being essential for functionality of ApbC in Fe-S protein biogenesis. However, the structural basis underlying the ApbC activity and the motifs' role remains unclear. Here, we report the crystal structure of Escherichia coli ApbC at 2.8 Å resolution. The dimeric structure is in a W shape and the active site is located in the 2-fold center. The function of the motifs can be annotated by structural analyses. ApbC has an additional N-terminal domain that differs from other P-loop NTPases, possibly conferring its inherent specificity in vivo.

The toxic effects of tetracycline exposure on the physiological homeostasis of the gut-liver axis in grouper.

Antibiotic residues, such as tetracycline (TET), in aquatic environments have become a global concern. The liver and gut are important for immunity and metabolism in aquatic organisms. In this study, juvenile groupers were subjected to 1 and 100 µg/L TET for 14 days, and the physiological changes of these fish were evaluated from the perspective of gut-liver axis. After TET exposure, the liver showed histopathology, lipid accumulation, and the elevated ALT activity. An oxidative stress response was induced in the liver and the metabolic pattern was disturbed, especially pyrimidine metabolism. Further, intestinal health was also affected, including the damaged intestinal mucosa, the decreased mRNA expression levels of tight junction proteins (ZO-1, Occludin, and Claudin-3), along with the increased gene expression levels of inflammation (IL-1ß, IL-8, TNF-α) and apoptosis (Casp-3 and p53). The diversity of intestinal microbes increased and the community composition was altered, and several beneficial bacteria (Lactobacillus, Bacteroidales S24-7 group, and Romboutsia) and harmful (Aeromonas, Flavobacterium, and Nautella) exhibited notable correlations with hepatic physiological indicators and metabolites. These results suggested that TET exposure can adversely affect the physiological homeostasis of groupers through the gut-liver axis.

[Evaluation of chemical constituents of Draconis Sanguis and its protective effect on renal tubular injury in diabetic kidney disease].

The chemical constituents of Draconis Sanguis were preliminarily studied by macroporous resin, silica gel, dextran gel, and high-performance liquid chromatography. One retro-dihydrochalcone, four flavonoids, and one stilbene were isolated. Their chemical structures were identified as 4-hydroxy-2,6-dimethoxy-3-methyldihydrochalcone(1), 4'-hydroxy-5,7-dimethoxy-8-methylflavan(2), 7-hydroxy-4',5-dimethoxyflavan(3),(2S)-7-hydroxy-5-methoxy-6-methylflavan(4),(2S)-7-hydroxy-5-methoxyflavan(5), and pterostilbene(6) by modern spectroscopy, physicochemical properties, and literature comparison. Compound 1 was a new compound. Compounds 2 and 6 were first found in the Arecaceae family. Compound 5 had the potential to prevent and treat diabetic kidney disease.

Effects of Cryptocaryon irritans infection on the histopathology, oxidative stress, immune response, and intestinal microbiota in the orange-spotted grouper Epinephelus coioides.

Cryptocaryon irritans is a parasitic ciliate of marine fish, causing serious mortality and economic loss of grouper. In this study, the orange-spotted grouper (Epinephelus coioides) were separately exposed to C. irritans infection for 72 h at a dose of 5000 or 10000 active theronts per fish, and we evaluated the changes in histopathology, oxidative stress, immune response, and intestinal microbiota composition. The results showed that C. irritans infection caused pathological alteration on the skin, gills, and liver of E. coioides. Oxidative stress responses occurred in the liver and gills, reflected in the corresponding antioxidant enzyme and gene indexes. The mRNA expression levels of inflammation-related genes (IL-1ß, IL-6, and IL-8) and the mediators of apoptosis (casp3, casp9, and cytc) were increased in the liver and gills of the fish. C. irritans infection also affected the diversity and composition of intestinal microbiota. Specifically, the relative abundance of Firmicutes was increased, whereas that of Proteobacteria was decreased. Several potentially beneficial bacteria (Pandoraea, Clostridium sensu stricto 1, Christensenellaceae R-7 group, and Weissella) were decreased, whereas pathogenic bacteria (Streptococcus and Acinetobacter) were increased. In conclusion, this study reveals that C. irritans infection caused histopathology, immune disorders, and intestinal microbial community variation in E. coioides.

Vibrio harveyi co-infected with Cryptocaryon irritans to orange-spotted groupers Epinephelus coioides.

The orange-spotted grouper (Epinephelus coioides) is a high economic value aquacultural fish in China, however, it often suffers from the outbreak of parasitic ciliate Cryptocaryon irritans as well as bacterium Vibrio harveyi which bring great loss in grouper farming. In the present study, we established a high dose C. irritans local-infected model which caused the mortality of groupers which showed low vitality and histopathological analysis demonstrated inflammatory response and degeneration in infected skin, gill and liver. In addition, gene expression of inflammatory cytokines was detected to assist the estimate of inflammatory response. Furthermore, we also found that the activity of Na+/K+ ATPase in gill was decreased in groupers infected C. irritans and the concentration of Na+/Cl- in blood were varied. Base on the morbidity symptom occurring in noninfected organs, we hypothesized that the result of morbidity and mortality were due to secondary bacterial infection post parasitism of C. irritans. Moreover, four strains of bacteria were isolated from the infected site skin and liver of local-infected groupers which were identified as V. harveyi in accordance of phenotypic traits, biochemical characterization and molecular analysis of 16S rDNA genes, housekeeping genes (gyrB and cpn60) and species-specific gene Vhhp2. Regression tests of injecting the isolated strain V. harveyi has showed high pathogenicity to groupers. In conclusion, these findings provide the evidence of coinfections with C. irritans and V. harveyi in orange-spotted grouper.

An Optimized Ni-catalyzed Chan-Lam Type Coupling: Enantioretentive Access to Chiral N-Aryl Sulfinamides.

C-N bond formation takes on a critical significance in reactions of organic synthesis, material production and pharmaceutical manufactory. Chan-Lam has proposed a useful methodology to furnish secondary arylamides under mild conditions. However, when chiral sulfinamides serve as the coupling precursors, the Cu-catalyzed coupling reaction is found with low efficacy. Complex side-products are generated under classic conditions. Moreover, it led to the racemization of the coupling product. In this study, an optimized Ni-catalyzed Chan-Lam type coupling conditions were proposed, which resulted in clean conversion from chiral sulfinamides and arylboronic acids to offer N-aryl sulfinamides efficiently and enantioretentively. The trans-N1 ,N2 -dimethylcyclohexane-1,2-diamine was proven as the most efficient ligand. Under the optimized conditions, a series of chiral N-aryl sulfinamides was prepared with high chemical yield without racemization. Furthermore, a plausible and novel mechanism was proposed. Interestingly, the method could efficiently furnish a wide variety of C-X bonds by coupling arylboronic acids with different nucleophiles.

Production of monoclonal antibody against grouper (Epinephelus coioides) CD4-1 and the distribution of CD4-1+ cells.

CD4-a transmembrane glycoprotein molecule expressed on the surface of helper T (Th) cells-plays a central role in adaptive immune protection. In the current study, we developed a monoclonal antibody (mAb) against the grouper CD4-1. Western blotting and immunohistochemistry results revealed that the CD4-1 mAb could recognize the recombinant and natural protein of grouper CD4-1 as well as the CD4-1+ cells in the various tissues from grouper. Tissue distribution analyses revealed that the grouper CD4-1+ cells were expressed in all tissues tested in the healthy grouper, with greater localization in the thymus, head kidney, and spleen tissues. In addition, we tested the changes in the proportion of CD4-1+ cells in the thymus, head kidney, and the gills of grouper post the infection by C. irritans. Our data suggest that the CD4-1 mAb produced against grouper in the current study can be used as a tool to characterize CD4-1+ cells and to investigate the functions of the grouper CD4-1+ cells in the host response against pathogens infection.

Essential oil of Magnolia denudata is an effective anesthetic for spotted seabass (Lateolabrax maculatus): a test of its effect on blood biochemistry, physiology, and gill morphology.

Magnolia denudata is a well-known ornamental tree in China due to its beautiful blossoms, and it has been used as an analgesic to treat human headaches. This study investigated the anesthetic potential and physiological response of the essential oil of M. denudata flowers on spotted seabass Lateolabrax maculatus. Fish (mean ± SD, 164.16 ± 15.40 g) were individually exposed to different concentrations of M. denudata essential oil (MDO, 10, 20, 40, 60, 80, 100, and 120 mg/L) and eugenol (10, 20, 30, 40, 50, 60, and 70 mg/L) to investigate anesthetic efficacy. Based on the ideal time criterion for anesthetic induction (< 3 min) and recovery (< 10 min), the lowest effective concentration for spotted seabass was 100 mg/L for MDO and 60 mg/L for eugenol. The physiological and histopathological damage in the gill of L. maculatus after using MDO and eugenol was also evaluated at the minimum dose inducing deep anesthesia, and at 0, 6, and 24 h after recovery. The results showed that MDO and eugenol anesthesia alleviated the levels of cortisol and glucose and the lactic dehydrogenase activity induced by handling. Compared with eugenol, MDO also caused secondary stress to the body, but MDO caused minor physiological responses and histological changes in the gills. This study suggests that MDO is an effective anesthetic for spotted seabass.

Characterization and functional study of nuclear factor erythroid 2-related factor 2 (Nrf2) in black tiger shrimp (Penaeus monodon).

Nuclear factor erythroid 2-related factor 2 (Nrf2) is a member of the Cap'n'collar basic region leucine zipper (CNC-bZIP) transcription factor family, and is activated by diverse oxidants, pro-oxidants, antioxidants and chemopreventive agents. The full-length cDNA of Nrf2 from Penaeus monodon (PmNrf2; 2024 bp long with 729 bp coding region, GenBank accession no. MW390830) was cloned. The 242-amino-acid polypeptide encoded by this gene had a predicted molecular mass of 27.80 kDa. Sequence homology and phylogenetic analysis showed that PmNrf2 was similar to the insect Cap'n'Collar (CNC) transcription factor and mammalian Nrf2. Tissue expression profile analyzed by quantitative real-time RT-PCR (qRT-PCR) demonstrated that PmNrf2 was constitutively expressed in all examined tissues, with the highest expression observed in the intestines and the weakest expression observed in the hemocyte. PmNrf2 expression profiles were detected in the hepatopancreas of shrimp after bacterial challenge. The results suggested that PmNrf2 was involved in the responses to bacterial challenge, but the temporal expression pattern trend of PmNrf2 differed between the gram-negative and gram-positive bacterial challenges in the shrimp hepatopancreas. The recombinant PmNrf2 protein was expressed and purified through affinity chromatography. Furthermore, an anti-PmNrf2 polyclonal antibody was obtained, which was able to clearly detect PmNrf2 protein expression in the hepatopancreas of shrimp. Knockdown of PmNrf2 by RNA interference (RNAi) resulted in a reduction in the expression of PmGPx gene. Taken together, the results of our study indicated that PmNrf2 played a role in regulation the transcription of PmGPx antioxidant enzyme genes.

Transcriptomic analysis of juvenile Chinese sea bass (Lateolabrax maculatus) anesthetized by MS-222 (tricaine methanesulfonate) and eugenol.

MS-222 (tricaine methanesulfonate) and eugenol are the two frequently used fish anesthetics. This study intends to analyze the regulation of these anesthetics in Chinese sea bass, Lateolabrax maculatus, through transcriptomic analysis. L. maculatus were exposed to MS-222 or eugenol, and those without any treatments were regarded as controls. Gills and livers were extracted for transcriptomic analysis after recovery in fresh water for 6 h. Identified genes were assigned to NR, COG, SWISS, GO, and KEGG database for predicting gene functions. A FDR ≤ 0.05 and |log2(FC)| ≥ 1 were applied to determined differentially expressed gene (DEG). A total of 45,626 unigenes were annotated using at least one database. The eugenol-treated liver group presented less DEGs compared with that treated by MS-222. Both the MS-222- and eugenol-treated liver groups presented notable DEGs that participated in human disease and metabolism pathways. The eugenol group showed more pathways related to detoxification activity and xenobiotics biodegradation, and those from the MS-222 group were related to organismal system such as reproduction. By comparing gill and liver samples using the same drug, the enriched pathways were generally consistent among the three comparisons. In conclusion, eugenol and MS-222 could change the pathways related to metabolism and immunity in L. maculatus. MS-222 may trigger more damages on the fish liver and reproduction.

Protection of teprenone against hypoxia and reoxygenation stress in stomach and intestine of Lateolabrax maculatus.

Teprenone (geranylgeranylacetone) is one kind of safe and effective agent in gastrointestinal mucosa, which have been widely used in human and veterinary, but rarely used in aquaculture animals. In this study, Lateolabrax maculatus, an important economic fish species in southern China, was taken as the object of study to investigate the protective effect of teprenone on intestinal stress. The present study was designed to investigate the potential mechanism underlying the protection offered by teprenone to protect the gastrointestinal tract against hypoxia and reoxygenation injury of L. maculatus. (a) For oxidative stress parameters, SOD, CAT, and T-AOC in control group were higher than those in teprenone group. MDA content was significantly higher than that in teprenone group at N and 12h time points in intestine (P < 0.05), and at 12, 24, and 48 h time points in stomach. (b) For immune-associated proteins, LZM activity in the control group was lower than that in the teprenone group, and the difference between the two groups in stomach and intestine was significant at 12.48 h and 6.48 h time points, respectively (P < 0.05). Compared with time point N, the content of HSP70 in the control group increased at 0 h in intestine. At 0-48 h, intestine HSP70 content in the control group showed a gradually decreasing trend, which was higher than that in the teprenone group. (c) For apoptosis-related factors, the activity of Cyt-C, caspase9, and caspase3 increased first and then decreased in both groups. The content of Cyt-C in the control group was significantly higher than that in the teprenone group at N-3.6 h, and 3.48 h time points in stomach and intestine, respectively (P<0.05). The activity of caspase9 and caspase3 was higher than that in the teprenone group at N-48 h. Results indicated that acute hypoxia and reoxygenation cause the expression levels of oxidative stress and apoptosis-related factors in the stomach and intestine increased first and then decreased within 0-48 h. Acute hypoxia and reoxygenation also that causes the level of nonspecific immunity decreased first and then increased. A total of 400-mg/kg treatment of teprenone can protect stomach and intestinal tissues to a certain extent. It can effectively protect oxidative stress and apoptosis within 0-48 h after acute hypoxia and reoxygenation and enhance non-specific immunity.

Correction to: The effect of teprenone on the intestinal morphology and microbial community of Chinese sea bass (Lateolabrax maculatus) under intermittent hypoxic stress.

In the end of the Discussion section, following information should be included.

The effect of teprenone on the intestinal morphology and microbial community of Chinese sea bass (Lateolabrax maculatus) under intermittent hypoxic stress.

Hypoxia stress may affect the fish intestine and thereby threaten the growth and survival of the fish. Teprenone is a clinically effective agent in protecting gastrointestinal mucosa. This study aims to assess the effect of teprenone in the intestine of Chinese sea bass Lateolabrax maculatus under intermittent hypoxic stress. L. maculatus juveniles were either raised under intermittent hypoxic condition or normal condition (NC). Part of the hypoxic-intervened fish were treated with teprenone at different concentrations (HTs), and the rest were regarded as hypoxic control (HC). Histological analysis was performed on the epithelial tissue of the fish intestine. High-throughput sequencing technology was used to analyze the diversity and composition of the microbial community in L. maculatus intestine. Reduced villi length and goblet cell, exfoliated enterocyte, and improper arrangement of villi were observed in HC compared with NC and HTs. Proteobacteria, Firmicutes, and Bacteroidetes represented the most abundant phyla in each sample. Significantly higher microbial diversity was detected in HC compared with NC (P < 0.05). At the phylum level, HC presented significantly decreased relative abundance of Proteobacteria, and significantly increased relative abundance of Bacteroidetes, Chloroflex, and Cyanobacteria compared with NC (P < 0.05). At the class level, HC showed significantly reduced relative abundance of Alphaproteobacteria and Bacilli, and significantly increased relative abundance of Clostridia, Gammaproteobacteria, and Bacteroides (P < 0.05). Teprenone protects the intestine from epithelial damages and maintains the microbial harmony in L. maculatus under intermittent hypoxic stress.

RNA-seq revealed the signatures of immunity and metabolism in the Litopenaeus vannamei intestine in response to dietary succinate.

The intestine is important for nutrition, metabolism and immunity. Succinate (SA) plays a vital role in the physiological homeostasis of animal intestines. However, the effects of dietary SA on the intestinal immunity and metabolism in shrimp are not clear. In this study, we investigated the immune and metabolic responses in the intestine of Litopenaeus vannamei that were fed diets consisting of different levels of SA: 0 g/kg (Con) and 10 g/kg (SA) for 56 days. The results from a RNA-seq analysis identified 6005 differentially expressed genes (DEGs), including 2728 upregulated genes and 3277 downregulated genes, which were grouped into 312 pathways. The DEGs were most enriched in pathways related to protein synthesis and amino acid metabolism, including "ribosome", "aminoacyl-tRNA biosynthesis", "pyrimidine metabolism", and "arginine and proline metabolism"; additionally, carbohydrate and lipid metabolism pathways were also activated. A large number of immune-related genes were associated with mucus barrier modification, antimicrobial activity, pathogen attachment and recognition, antioxidant activity, and apoptosis. The expression patterns of several candidate genes involved in the immune response and nutrition metabolism were detected by qPCR. This study provides insight into the transcriptomic modulating mechanisms associated with intestinal immunity and the metabolism of L. vannamei in response to the intake of dietary SA.

Changes in the intestine barrier function of Litopenaeus vannamei in response to pH stress.

pH of water environment affects the survival of aquatic animals. Intestine barrier function influences the health of animals, which is related to its mucosa structure, immune components, and microbial communities. In this study, we investigated the histological structure, digestive and metabolic capacity, immune responses, and microbial composition in the intestine of Litopenaeus vanmei under three different conditions: control (pH 8.3), low pH stress (pH 6.9), and high pH stress (pH 9.7) for 72 h. The results showed both low and high pH stress disrupted the intestine morphological structure, and induced variations in the activities of digestive (AMS, LPS, Tryp, and Pep) and metabolic (HK, PK, CCO, and LDH) enzymes. Low and high pH stress also increased oxidative stress (MDA, LPO, PC, and ·O2- generation), and decreased the antioxidant enzyme activities (T-AOC, SOD, and GST); shrimp enhanced CAT activity and HSP70, Trx, MT and Fer gene transcripts as defense mechanism. Additionally, Immune confusion was also found in the shrimp intestine in response to low and high pH stress, including the antibacterial ability (T-NOS, PO, proPO, ALF, and Lys), pathogen recognition (TLR and Lec), apoptosis (Casp, IAP and p53), and mucus homeostasis (Muc-1, Muc-2, Muc-5AC, Muc-5B, and Muc-19). pH exposure also decreased the diversity of the intestine bacterial, disturbed the composition of microbiota, and decreased the microbial metabolite SCFA contents. Our results indicated that acute pH stress can impair the intestine barrier function of white shrimp, probably via destroying mucosa structure, confusing digestion and metabolism, inducing oxidative stress, disordering immunity, and disrupting the microbial composition.

Transcriptomic and microbiota response on Litopenaeus vannamei intestine subjected to acute sulfide exposure.

Harmful effects of water pollutants are myriad. Sulfide from water bodies affects the aquatic animals. Intestine barrier function serves as the front-line of animals defense. Our previous study confirmed the toxic effect of sulfide on intestine immune response of Litopenaeus vannamei, but the underlying mechanisms remained elusive. Therefore, in this study, we investigated the transcriptomic and microbiota responses of the L. vannamei intestine subjected to acute sulfide exposure. Sulfide decreased bacterial richness and altered the intestine microbiota composition. Specifically, sulfide increased the abundances of Bacteroidetes and Actinobacteria, but decreased the abundance of Proteobacteria. At the genus level, sulfide increased typical cellulolytic characteristics bacteria, such as Formosa, Sphingomonas, and Demequina. RNA-seq analysis identified differential expression of 1799 genes (701 up-regulated and 1098 down-regulated) were grouped into 267 pathways. The most enriched pathway 'amoebiasis' was related to the intestine mucus homeostasis. A number of immune-related genes associated with antimicrobial, antioxidant, pathogen attachment and recognition, and apoptosis processes in contrasting accessions; they were correlated with the abundance of intestine bacterial at the phylum level. This study provides an insight into the mechanisms associated with molecular and microbiota response and processes involved in adaptation strategies towards sulfide stress.

Effects of palytoxins extracted from Ostreopsis ovata on the oxidative stress and immune responses in Pacific white shrimp Litopenaeus vannamei.

Palytoxins (PLTXs) are a group of complex and poisonous marine natural products that are toxic to marine life and even human beings. In the present study, the oxidative stress and immune response in the hepatopancreas and gills of Litopenaeus vannamei were assessed for 72 h after injection with PLTX extracts. Chemical and physiological parameters, e.g., the respiratory burst (O2-), activities of antioxidant enzymes, oxidative damage to lipids, carbonylation of proteins, and immune gene mRNA expression levels, were analysed. The results showed that the PLTX extract was not fatal to the shrimp but could reduce their mobility. The O2- levels in the gills gradually increased after exposure to PLTX extracts and were significantly higher than those in the control from 6 to 72 h. The malondialdehyde content, lipid peroxidation, protein carbonyl levels, and total antioxidant capacity in the gills all peaked at 12 h. At the same time, the gills were loosely connected, there was a clear disintegration of the epithelial tissue, and the stratum corneum disappeared after 12 h. In addition, compared to those in the control group, the PLTX extract treatment increased the O2- content, malondialdehyde content, lipid peroxidation, and protein carbonyl levels from 12 to 72 h, 24-48 h, 12-24 h, and 12-72 h after injection in the hepatopancreas of the shrimp, respectively. Both the Crustin and Toll gene expression levels significantly increased in the hepatopancreas compared to those in the control 6-72 h after injection of the toxin. In parallel, the expression levels of the manganese superoxide dismutase gene gradually decreased from 6 to 48 h and returned to normal levels after 72 h. Interestingly, the total antioxidant capacity also significantly increased compared to that in the control from 6 to 72 h. Our results indicate that although PLTX extracts cause lipid peroxidation and carbonylation of proteins in hepatopancreatic cells, leading to their damage, they did not cause a decrease in the total antioxidant capacity of the hepatopancreas.

Impairment of the intestine barrier function in Litopenaeus vannamei exposed to ammonia and nitrite stress.

Intestine barrier serves as the front-line of shrimp defense, which rely on its structural integrity, microbial composition, and mucus immune compounds. Mucins are the major organic components of the intestine mucus layer that contribute to the immunity of intestine mucus. In this study, we examined the histological structure, microbial composition, and mucin genes expression in the intestines of Litopenaeus vanmei under three different conditions: control, ammonia stress, and nitrite stress for 72 h. H&E stain showed that ammonia and nitrite stress exposure both damaged the intestine mucosal tissue. High-throughput 16S rDNA sequencing revealed that two stresses exposure decreased the bacterial diversity, and altered the composition of intestine microbial. Specifically, the dominant bacterial phyla Bacteroidetes abundance was increased, while Proteobacteria and Planctomycetes were decreased; at the genus level, Formosa abundance was increased and Photobacterium was decreased, opportunistic pathogens including Nautella and Pseudoalteromonas was also increased. Intestine mucus immune genes including mucin-2 and mucin-19 were up-regulated, while mucin-1, mucin-5AC, and mucin-5B were down-regulated in two stress exposure groups. These results revealed that ammonia and nitrite stress harmed the intestine barrier function of L. vannamei by damaging the mucosal tissue, disrupting the composition of intestine microbial, and suppressing the immune function.

Dietary effects of succinic acid on the growth, digestive enzymes, immune response and resistance to ammonia stress of Litopenaeus vannamei.

Organic acids acts as an growth promoter and antimicrobial agent in aquaculture. The present study investigated the effects of a natural organic acid - succinic acid (SA) on the growth, digestive enzymes, immune response and resistance to ammonia stress of Litopenaeus vannamei. The shrimps were firstly fed with diets containing different levels of SA: 0% (Control), 0.25% (SA1), 0.50% (SA2), and 1.0% (SA3) (w/w) for 56 days, followed by an acute ammonia stress for 48 h. The results indicated that dietary of SA improved the growth of shrimp, and increased the survival rate of shrimp after ammonia stress for 48 h. The amylase, lipase and pepsin activity increased in hepatopancreas in three SA group, while trypsin activity was only increased in the SA1 and SA2 groups. At 56 d, T-NOS activity, proPO and HSP70 gene expression level increased in the three SA group, PO activity increased in the SA1 and SA2 groups, T-AOC content and Toll gene expression level increased in the SA2 and SA3 groups, Trx and SOD gene expression level increased in the SA2 group, while Imd, GS and GDH gene expression level was no changes. After exposure to ammonia stress for 48 h, immune biochemical parameters (T-AOC and PO) and genes (proPO, HSP70, Trx and GDH) expression level increased in the three SA group, T-NOS activity, Toll, Imd and GS gene expression level increased in the SA2 and SA3 groups, while SOD gene expression level increased in the SA1 and SA2 groups. These results indicated that SA improved growth, enhanced digestive and immune capacities of L. vannamei against ammonia stress, and may be a potential feed additive for shrimp. The optimal dietary supplementation dosage is 0.50% (w/w) in diet.

Physiological and immune response in the gills of Litopenaeus vannamei exposed to acute sulfide stress.

Sulfide is a harmful environmental pollutant that affects the survival and immunity of shrimps. The gill is important for shrimp respiratory and osmotic adjustment, the physiological and immune homeostasis of the organ can be influenced by sulfide. In this study, we investigated the acute toxicity of sulfide (5 mg/L) on the morphology, physiological and immune response in the gills of Litopenaeus vannamei. H&E stain showed that sulfide stress damaged the gills histological structure. Specifically, osmoregulation capacity including of Na+/K+-ATPase and Ca2+/Mg2+-ATPase activity was both increased at 6 h and 12 h, and decreased at 72 h; the contents of free amino acid including of Gly, Pro, Ser, Thr and Ala were decreased at 72 h. Respiratory metabolic enzymes, such as cytochrome c oxidase and succcinate dehydrogenase activity was decreased at 12 h-72 h, while fumarate reductase and lactate dehydrogenase activity kept a higher level at 12 h-72 h. Significant variations in the activities of immune enzymes (acid phosphatase, alkaline phosphatase, total antioxidant capacity and lysozyme). The expression of immune-related genes (heat shock protein 70, thioredoxin and caspase-3) was increased at first and then decreased, while hypoxia inducible factor 1α kept a higher level at 6 h-72 h. These results revealed that sulfide stress influenced the L. vannamei gills physiological and immune function by damaging histological structure, and confusing osmoregulation, respiratory metabolic and immune capacity.