Insight into three water additives: Revealing the protective effects on survival and stress response under cold stress for Pacific white shrimp Litopenaeus vannamei.

The reproduction, development and growth of shrimp were hindered by cold stress, and even death was caused in severe cases. Moreover, huge economic losses to the shrimp aquaculture industry were caused every year by cold currents. The purpose of this study was to investigate the potential protective effects of water additives on the cold stress resistance of Pacific white shrimp (Litopenaeus vannamei) and their ability to improve the survival and stress response of the shrimp. Three potential cold-resistant additives adenosine triphosphate (A), soybean phospholipid (SP) and Clostridium butyricum (CB) on Pacific white shrimp under cold stress were added to the water with three concentrations for each additive. The mortality, activities of antioxidation enzymes and expression of anti-stress related genes in each group under cold stress were detected. The results showed that the cumulative mortality of low concentration for adenosine triphosphate (AL) and soybean phospholipid (SPL), medium concentration for soybean phospholipid (SPM) and high concentration for Clostridium butyricum (CBH) groups were significantly lower than that of the control (C) group when temperature maintained at 13 °C for 6 days. Total antioxidant capacity (T-AOC) content in shrimp plasma was significantly higher, while malondialdehyde (MDA) content was significantly lower than that in the C group. Gene expression analysis showed that 0.4 mg/L of adenosine triphosphate could regulate the immune defense ability and decrease apoptosis level of Pacific white shrimp under cold stress. Soybean phospholipid (2 mg/L) could enhance the immune ability of hepatopancreas, and Clostridium butyricum (10 mg/L) could significantly increase the expression of stress-related genes in shrimp intestine. Overall, these findings suggested that adenosine triphosphate and soybean phospholipid have the potential to be used as cold-resistant additives in Pacific white shrimp culture. This study provided valuable insights into addressing the problem of cold stress in shrimp culture.

New insights into the regulation mechanism of Litopenaeus vannamei hepatopancreas after lipopolysaccharide challenge using transcriptome analyses.

Litopenaeus vannamei (L. vannamei) is the most economically valuable cultured shrimp in the world, while Gram-negative bacteria infection causes huge economic losses to shrimp culture. In this study, we performed transcriptome sequencing of the hepatopancreas in L. vannamei after lipopolysaccharide (LPS, the cell wall component of Gram-negative bacteria) injection to investigate the response of shrimp under Gram-negative bacteria invasion. A total of 306 differentially expressed genes (DEGs) (70 up- and 236 down-regulated) were identified in the LPS treatment group (L group) when compared to their expression levels in the control group (C group). The oxidoreductase activity (GO:0016491) in the molecular function category was enriched in the LPS-responsive DEGs in GO annotation, and the metabolism of xenobiotics by cytochrome P450 (ko00980) was the most enriched pathway in KEGG annotation. The transcriptome profiling revealed that the toll like receptor, C-type lectin receptor, and ß-1,3-glucan binding protein were involved in the recognition of LPS during its early invasion stage. Although LPS could reduce the metabolic ability of exogenous substances, induce inflammation and reduce antioxidant capacity, L. vannamei could maintain its homeostasis by improving immunity, enhancing anti-stress ability and reducing apoptosis. Our research provides the first transcriptome profiling for the L. vannamei hepatopancreas after LPS injection. These results could offer a valuable reference on the mechanism of shrimp against Gram-negative bacteria and could provide guidance for shrimp farming.

Intestinal morphology and microflora to Vibrio alginolyticus in pacific white shrimp (Litopenaeus vannamei).

In recent years, the shrimp farming industry encountered significant economic losses induced by Vibrio alginolyticus. In this study, the influence of Vibrio alginolyticus on intestinal histomorphology and microbiome composition in Litopenaeus vannamei were studied. The results showed that the intestinal mucosal epithelial cells of Vibrio group (VA group) injected only with Vibrio alginolyticus showed large area exfoliation at 12 h, and the tissue morphology of intestine recovered at 48 h. Compared with the control group (CK group), the abundance of Proteobacteria was significantly higher (P < 0.05), while the abundance of Actinobacteria was significantly lower after infection with Vibrio alginolyticus. The abundance of Shewanella in intestinal microbiome of Litopenaeus vannamei was significantly higher at 12 h (P < 0.05), but the abundance of Candidatus_Bacilloplasma was significantly lower at 48 h after infection (P < 0.05). In VA group, the diversity of intestinal microbiome was significantly lower at 12 h, which could be caused by the proliferation of Candidatus_Bacilloplasma and Shewanella. All above findings suggested that the stability of the dynamic balance of microbiome in the intestine helped Litopenaeus vannamei to resist pathogen colonization.

Comparative analysis on the immunoregulatory roles of ferritin M in hybrid fish (Carassius cuvieri ♀ × Carassius auratus red var ♂) and its parental species after bacterial infection.

Ferritin M is involved in the regulation of fish immunity. In this study, open reading frame (ORF) sequences of ferritin M from hybrid fish and its parental species were 534 bp. Tissue-specific analysis indicated that the highest level of ferritin M from red crucian carp was observed in kidney, while peaked expressions of ferritin M from white crucian carp and hybrid carp were observed in gill. Elevated levels of ferritin M from hybrid carp and its parental species were detected in immune-related tissues following Aeromonas hydrophila infection or in cultured fish cell lines after lipopolysaccharide (LPS) challenge. Ferritin M overexpression could attenuate NF-κB and TNFα promoter activity in their respective fish cells. Purified ferritin M fusion proteins elicited in vitro binding activity to A. hydrophila and Edwardsiella tarda, lowered bacterial dissemination to tissues and alleviated inflammatory response. Furthermore, treatment with ferritin M fusion proteins could mitigate bacteria-induced liver damage and rescue antioxidant activity. These results suggested that ferritin M in hybrid fish showed a similar immune defense against bacteria infection in comparison with those of its parental species.

Blood cell characterization and transcriptome analysis reveal distinct immune response and host resistance of different ploidy cyprinid fish following Aeromonas hydrophila infection.

Aeromonas hydrophila can pose a great threat to survival of freshwater fish. In this study, A. hydrophila infection could decrease blood cell numbers, promote blood cell damage as well as alter the levels of alkaline phosphatase (ALP), lysozyme (LZM), aspartate aminotransferase (AST), total antioxidant capacity (T-AOC), total superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in immune-related tissues of red crucian carp (RCC, 2 N = 100) and triploid cyprinid fish (3 N fish, 3 N = 150). In addition, the significant alternation of antioxidant status was observed in PBMCs isolated from RCC and 3 N following LPS stimulation. The core differential expression genes (DEGs) involved in apoptosis, immunity, inflammation and cellular signals were co-expressed differentially in RCC and 3 N following A. hydrophila challenge. NOD-like receptor (NLR) signals appeared to play a critical role in A. hydrophila-infected fish. DEGs of NLR signals in RCCah vs RCCctl were enriched in caspase-1-dependent Interleukin-1ß (IL-1ß) secretion, interferon (IFN) signals as well as cytokine activation, while DEGs of NLR signals in 3Nah vs 3Nctl were enriched in caspase-1-dependent IL-1ß secretion and antibacterial autophagy. These results highlighted the differential signal regulation of different ploidy cyprinid fish to cope with bacterial infection.

Transcriptome analysis and co-expression network reveal the mechanism linking mitochondrial function to immune regulation in red crucian carp (Carassius auratus red var) after Aeromonas hydrophila challenge.

Aeromonas hydrophila is a common pathogen of freshwater fish. In this study, A. hydrophila infection was shown to cause tissue damage, trigger physiological changes as well as alter the expression profiles of immune- and metabolic-related genes in immune tissues of red crucian carp (RCC). Transcriptome analysis revealed that acute A. hydrophila infection exerted a profound effect on mitochondrial oxidative phosphorylation linking metabolic regulation to immune response. In addition, we further identified cellular senescence, apoptosis, necrosis and mitogen-activated protein kinase signal pathways as crucial signal pathways in the kidney of RCC subjected to A. hydrophila infection. These findings may have important implications for understanding modulation of immunometabolic response to bacterial infection.

Comparative analysis of erythrocyte hemolysis, plasma parameters and metabolic features in red crucian carp (Carassius auratus red var) and triploid hybrid fish following Aeromonas hydrophila challenge.

Aeromonas hydrophila can pose a great threat to survival of freshwater fish. In this study, A. hydrophila challenge could promote the erythrocyte hemolysis, increase free hemoglobin (FHB) level and generate malondialdehyde (MDA) production in plasma but decrease the levels of total antioxidant capacity (T-AOC), total superoxide dismutase (SOD), catalase (CAT), alkaline phosphatase (ALP) and lysozyme (LZM) of red crucian carp (RCC, 2 N = 100) and triploid hybrid fish (3 N fish, 3 N = 150) following A. hydrophila challenge. Elevated expression levels of heat shock protein 90 alpha (HSP90α), matrix metalloproteinase 9 (MMP-9), free fatty acid receptor 3 (FFAR3), paraoxonase 2 (PON2) and cytosolic phospholipase A2 (cPLA2) were observed in A. hydrophila-infected fish. In addition, A. hydrophila challenge could significantly increase expressions of cortisol, leucine, isoleucine, glutamate and polyunsaturated fatty acids (PUFAs) in RCC and 3 N, while glycolysis and tricarboxylic acid cycle appeared to be inactive. We identified differential fatty acid derivatives and their metabolic networks as crucial biomarkers from metabolic profiles of different ploidy cyprinid fish subjected to A. hydrophila infection. These results highlighted the comparative metabolic strategy of different ploidy cyprinid fish against bacterial infection.

A novel NK-lysin in hybrid crucian carp can exhibit cytotoxic activity in fish cells and confer protection against Aeromonas hydrophila infection in comparison with Carassius cuvieri and Carassius auratus red var.

NK-lysin, an effector of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs), not only exhibits cytotoxic effect in fish cells, but also participates in the immune defense against pathogenic infection. In this study, ORF sequences of RCC-NK-lysin, WCC-NK-lysin and WR-NK-lysin were 369 bp. Tissue-specific analysis revealed that the highest expressions of RCC-NK-lysin and WCC-NK-lysin were observed in gill, while the peaked level of WR-NK-lysin mRNA was observed in spleen. A. hydrophila infection sharply increased RCC-NK-lysin, WCC-NK-lysin and WR-NK-lysin mRNA expression in liver, trunk kidney and spleen. In addition, elevated levels of NK-lysin mRNA were observed in cultured fin cell lines of red crucian carp (RCC), white crucian carp (WCC) and their hybrid offspring (WR) after Lipopolysaccharide (LPS) challenge. RCC-NK-lysin, WCC-NK-lysin and WR-NK-lysin exerted regulatory roles in inducing ROS generation, modulating mitochondrial membrane potential, decreasing fish cell viability and antagonizing survival signalings, respectively. RCC/WCC/WR-NK-lysin-overexpressing fish could up-regulate expressions of inflammatory cytokines and decrease bacterial loads in spleen. These results indicated that NK-lysin in hybrid fish contained close sequence similarity to those of its parents, possessing the capacities of cytotoxicity and immune defense against bacterial infection.

Impacts of microplastics on three different juvenile shrimps: Investigating the organism response distinction.

The effects of microplastics (MPs) on aquaculture animals have raised increasing concern, but studies on MPs contamination in cultured shrimp are still limited. Therefore, the responses of three widely farmed shrimp species to MPs, including Penaeus monodon (P. monodon), Marsupenaeus japonicas (M. japonicus) and Litopenaeus vannamei (L. vannamei), were investigated in this study. The results showed that the mortality of P. monodon, M. japonicus and L. vannamei were 47%, 53% and 20% respectively after 48 h of 300 mg/L MPs exposure. After 48 h of 100 mg/L MPs exposure, for P. monodon, the MPs content in water and excreta were significantly different from that in M. japonicus and L. vannamei. For genes expressions, the expression of catalase (Cat) was significantly increased and the expression of apoptosis protein (IAP) was inhibited in these three shrimps, but only the expression of Lysozyme (Lys) was increased in L. vannamei after MPs exposure. After 48 h of depuration, the Cat and IAP expression of P. monodon and M. japonicus was significant decreased while the IAP and Lys expression of L. vannamei still maintained at a high level. The results suggested that the metabolic rate of MPs in P. monodon was significantly higher than that in M. japonicus and L. vannamei. The tolerance of L. vannamei to MPs was higher than that of P. monodon and M. japonicas and their different responses in anti-microbial gene might be one of the reasons for the difference of their mortality. This study provides the first report comparing the organism response distinction in cultured shrimp and enriching to the understanding of the impact of MPs on ecosystem.

The pivotal protein profile between the conjoined twins and normal mosquitofish Gambusia affinis based on iTRAQ proteomic analysis.

The fish abnormal embryonic development has attracted public attention in the recent few years. In this study, an iTRAQ proteomic analysis of mosquitofish between conjoined twins and normal fishes is applied for the first time by using the genome database of mosquitofish. Three thousand four hundred ninety proteins were identified with 304 differentially expressed proteins (DEPs). One hundred six differentially upregulated proteins (DUPs) and 198 differentially downregulated proteins (DDPs) were identified between the conjoined twins and normal mosquitofish groups. Notably, the proteins related to lipid and proteolysis were the important GO terms for the DUPs while response to light stimulus and response to radiation were the most enriched GO terms for the DDPs. The proteins related to lysosome, apoptosis, autophagy, and phagosome were the functional KEGG pathway for the DUPs while most of the pathways were related to cardiovascular for the DDPs. This study expatiated a pivotal protein profile between the conjoined twins and normal mosquitofish which can provide a conference for fish embryonic development.

New insights into the immune regulation and tissue repair of Litopenaeus vannamei during temperature fluctuation using TMT-based proteomics.

To investigate shrimp immunoregulation and tissue self-repair mechanism during temperature fluctuation stage, Litopenaeus vannamei (L. vannamei) was treated under conditions of gradual cooling from an acclimation temperature (28 °C, C group) to 13 °C (T group) in 2 days with a cooling rate of 7.5 °C/d and then rewarmed to 28 °C (R group) with the same rate. Tandem mass tags (TMT) -based proteomics technology was used to investigate the protein abundance changes of intestine in L. vannamei during temperature fluctuation. The results showed that a total of 5796 proteins with function annotation were identified. Of which, the abundances of 1978 proteins (34%) decreased after cooling and then increased after rewarming, 1498 proteins (26%) increased during the whole stage, 1263 proteins (22%) increased after cooling and then decreased after rewarming and 1057 proteins (18%) decreased during the whole stage. Differentially expressed proteins such as C-lectin, NFκBIA and Caspase may contributed to the regulation of immunity and tissue repair of shrimp intestine during the temperature fluctuation stage. These findings contribute to the better understanding of shrimp' regulatory mechanism against adverse environment.

The immune defense response of Pacific white shrimp (Litopenaeus vannamei) to temperature fluctuation.

Temperature is a significant environmental factor contributing to the success of aquaculture. To investigate the immune defense response during temperature fluctuation, Litopenaeus vannamei (L. vannamei) was treated under conditions of gradual cooling from an acclimation temperature of 28 °C (C group) to 13 °C (T group) in 2 days with a cooling rate of 7.5 °C/d and then rewarmed to 28 °C (R group) using the same rate. Relative expression of immune defense system-related genes and intestinal microbial composition in L. vannamei were investigated. The results showed that with a decrease in temperature, the expression of TLR, IMD, proPO and Casp3 in intestine was significantly decreased, while the expression of Muc-3A, Muc-5AC, Muc-17, IAP, p53, HSP70, MT and Fer was significantly increased after cooling. After temperature recovery, gene expression generally showed a trend of recovering to the normal level (C). Intestinal microbial analysis showed that, compared with the C group, the Chao and Ace indexes, the relative abundance of microflora from the Phylum Bacteroidetes, Class Alphaproteobacteria, and Class Bacteroidia, significantly decreased in the R group. The results revealed that cold-stress may decrease microbial community richness, alter the bacterial community in general and reduce shrimp immunity to pathogens and antibacterial activity. As a result, during temperature fluctuation shrimp may mobilize the immune defense system through upregulating the expression of Muc genes, anti-apoptosis related genes, and antioxidant related genes in order to maintain organism homeostasis as well as to repair damaged intestinal tissue.

WSV152 induces apoptosis and promotes viral replication in Litopenaeus vannamei.

Previous studies have indicated that white spot syndrome virus (WSSV) infection induces apoptosis in many shrimp organs. However, the mechanism by which WSSV causes host apoptosis remains largely unknown. In this study, we demonstrated the function of wsv152, the first mitochondrial protein identified as encoded by WSSV. Glutathione S-transferase pulldown and co-immunoprecipitation analysis revealed that wsv152 interacts with the shrimp mitochondrial protein cytochrome c oxidase 5a (COX5a), a subunit of the COX complex. We also found that wsv152 expression significantly increased the rate of apoptosis, suggesting a role of wsv152 in WSSV-induced apoptosis in shrimp. Knockdown of wsv152 in vivo led to downregulation of several apoptosis-related shrimp genes, including cytochrome c, apoptosis-inducing factor and caspase-3. Suppression of wsv152 also resulted in significant reductions in the number of WSSV genome copies in tissues and in the mortality of WSSV-infected shrimp. Together, these results suggest that wsv152 targets host COX5a and is associated with the expression profiles of apoptosis-related shrimp genes. Wsv152 is likely also involved in WSSV-induced apoptosis, thereby facilitating virus infection and playing a complex role in WSSV pathogenesis.

Cytochrome P450 1A mRNA in the Gambusia affinis and Response to Several PAHs.

Cytochrome P4501A (CYP1A) has been used as a specific biomarker for monitoring water contamination such as PAHs, PCBs and dioxins. In the present study, the cyp1a gene of Gambusia affinis was cloned and sequenced and their expressions under PAHs exposure were characterized. The newly identified cyp1a encodes a protein with 521 amino acids that shared 96-80% identity with other Cyprinodontiformes fishes. RT-PCR analysis revealed that the basal mRNA level of cyp1a was highly expressed in liver and intestine. The expression level of cyp1a was significantly induced by exposure to 100 µg/L 3, 4-Benzopyrene (BaP) for 5 days in the muscle, testis, brain, liver and intestine of adult male fish. Except in the testis, the induced mRNA level of cyp1a ultimately decreased after prolonging the exposure time to 25 days. As for testis, the induced mRNA level of cyp1a was maintained at a high level during the entire exposure time under 100 µg/L BaP exposure. Furthermore, the expression of cyp1a increased with exposure time under a relatively low exposure concentrations 1 µg/L. Regarding the effects of other PAHs, D(a,h)A, BbF, and BaA showed a statistically significant effect of induction on mRNA level of cyp1a in the muscle, testis, brain, liver and intestine.

Proteomic characterization of the hepatopancreas in the Pacific white shrimp Litopenaeus vannamei under cold stress: Revealing the organism homeostasis mechanism.

To understand the homeostasis mechanism of crustacean hepatopancreas to cold stress, iTRAQ proteomics based on the genome database of Litopenaeus vannamei (L. vannamei) was applied to investigate proteins changes and variety of the hepatopancreas during cold stress stage in this study. A total of 4062 distinct proteins were identified, 137 differentially expressed proteins (DEPs) including 62 differentially up-regulated proteins (DUPs) and 75 differentially down-regulated proteins (DDPs) were identified in G1 (18 °C) compared with CK (28 °C), 359 DEPs including 131 DUPs and 228 DDPs were identified in G2 (13 °C for 24 h) compared with CK. Based on bioinformatics analysis, the cold tolerance of L. vannamei might be related to energy metabolism such as amino acid, carbohydrate, lipid, and oxidative phosphorylation. Moreover, shrimp immunity was declined during cold stress stage. However, L. vannamei could cope with cold stress by enhancing the production of ATP and UFA. Notably, arginine kinase, heat shock proteins, and histones may act as positive regulators in L. vannamei under cold stress. Ten randomly selected proteins were used for validation using qRT-PCR and the expressions on the transcription level for most of the genes were similar to the results of iTRAQ. These results indicated that L. vannamei can maintain the organism homeostasis by a series of orderly regulatory process during cold stress. Furthermore, the results can provide guidance for shrimp farming.

White spot syndrome virus (WSSV) infection impacts intestinal microbiota composition and function in Litopenaeus vannamei.

Intestinal microbiota homeostasis is crucial to the health of host. Pathogen invasion results in dynamics of microbiota composition and structure, disrupting their function in maintaining host health. WSSV is the most prevalent viral pathogen and is able to cause extremely high mortality in Litopenaeus vannamei. However, the changes of intestinal microbiota induced by WSSV are yet to be elucidated. In this study, we analyzed and compared the microbiota of healthy and WSSV-challenged shrimp intestines. Though the richness and diversity of microbiota was barely affected by WSSV, the abundance of predominant phyla like Proteobacteria and Fusobacteria were upregulated significantly, while Bacteroidetes and Tenericutes were significantly decreased in WSSV-infected shrimps. At the genus level, significant increase was observed in Photobacterium, Propionigenium and Arcobacter, as well as significant decrease in Candidatus Bacilloplasma and Flavobacterium in WSSV-infected shrimps. Additionally, metagenomic predictions by PICRUSt suggested that the altered microbiota was mainly related to metabolism, human diseases, genetic information processing, environmental information processing and cellular processes. These results suggested that the invasion of WSSV could impact intestinal microbiota composition and function in L. vannamei.

Fish SUMO3 functions as a critical antiviral molecule against iridovirus and nodavirus.

Protein SUMOylation (SUMO is small ubiquitin-related modifier) is a dynamic process that is strictly regulated under physiological and pathological conditions. We previously cloned and characterized two SUMO homologue genes (EcSUMO1 and EcSUMO2) from orange-spotted grouper (Epinephelus coioides). In the present study, the SUMO3 homologue from E. coioides (EcSUMO3) was cloned and its possible roles in fish immunity were analyzed. The open reading frame of EcSUMO3 contains 285 base pairs encoding a 94 amino acid protein with a predicted molecular mass of 10.73 kDa. The protein sequence of EcSUMO3 revealed similar domains with mammals, including the UBQ (ubiquitin-like proteins) domain, the hydrophobic surface, the Ulp1-Smt3 interaction sites, a VKTE motif and the C-terminal Gly residues. EcSUMO3 shares 46.83% and 89.58% identity with EcSUMO1 and EcSUMO2, respectively, and it shares 94%, 98%, and 98% identity with SUMO3 from Oreochromis niloticus, Danio rerio, and Homo sapiens, respectively. Quantitative real-time polymerase chain reaction analysis indicated that EcSUMO3 was constitutively expressed in all of the analyzed tissues in healthy grouper. EcSUMO3 expression levels were remarkably (p < 0.01) up-regulated in grouper spleen (GS) cells in response to stimulation with red-spotted grouper nervous necrosis virus (RGNNV) and Singapore grouper iridovirus (SGIV). EcSUMO3 was distributed in both the cytoplasm and nucleus in GS cells. EcSUMO3 enhanced SGIV and RGNNV replication during viral infection in vitro. These results are important for better understanding of the SUMO pathway in fish and provide insights into the regulatory mechanism of viral infection in E. coioides under farmed conditions.

Effects of dietary Gelsemium elegans alkaloids on intestinal morphology, antioxidant status, immune responses and microbiota of Megalobrama amblycephala.

Numerous plant extracts used as feed additives in aquaculture have been shown to stimulate appetite, promote growth and enhance immunostimulatory and disease resistance in cultured fish. However, there are few studies on the famous Chinese herbal medicine Gelsemium elegans, which attracts our attention. In this study, we used the Megalobrama amblycephala to investigate the effects of G. elegans alkaloids on fish intestinal health after diet supplementation with 0, 5, 10, 20 and 40 mg/kg G. elegans alkaloids for 12 weeks. We found that dietary G. elegans alkaloids at 40 mg/kg improved intestinal morphology by increasing villus length, muscle thickness and villus number in the foregut and midgut and muscle thickness in the hindgut (P < 0.05). These alkaloids also significantly improved intestinal antioxidant capabilities by increasing superoxide dismutase, catalase, total antioxidant capacity and malondialdehyde levels and up-regulated intestinal Cu/Zn-SOD and Mn-SOD (P < 0.05) at 20 and 40 mg/kg. Dietary G. elegans alkaloids improved intestinal immunity via up-regulating the pro-inflammatory cytokines IL-1ß, IL-8, TNF-α and IFN-α and down-regulating expression of the anti-inflammatory cytokines IL-10 and TGF-ß (P < 0.05) at 20 and 40 mg/kg. The expression of Toll-like receptors TRL1, 3, 4 and 7 were also up-regulated in intestine of M. amblycephala (P < 0.05). In intestinal microbiota, the abundance of Proteobacteria was increased while the Firmicutes abundance was decreased at phylum level after feeding the alkaloids (P < 0.05). The alkaloids also increased the abundance of the probiotic Rhodobacter and decreased the abundance of the pathogenic Staphylococcus at genus level (P < 0.05). In conclusion, dietary G. elegans alkaloid supplementation promoted intestine health by improving intestine morphology, immunity, antioxidant abilities and intestinal microbiota in M. amblycephala.

Integrated analysis of intestinal microbiota and transcriptome reveals that a coordinated interaction of the endocrine, immune system and gut microbiota response to heat stress in Litopenaeus vannamei.

Due to the ongoing global warming, the risk of heatwaves in the oceans is continuously increasing while our understanding of the physiological response of Litopenaeus vannamei under extreme temperature conditions remains limited. Therefore, this study aimed to evaluate the physiological responses of L. vannamei under heat stress. Our results indicated that as temperature rose, the structure of intestinal and hepatopancreatic tissues was damaged sequentially. Activity of immune-related enzymes (acid phosphatase/alkaline phosphatase) initially increased before decreased, while antioxidant enzymes (superoxide dismutase and glutathione-S transferase) activity and malondialdehyde content increased with rising temperature. In addition, the total antioxidant capacity decreased with rising temperature. With the rising temperature, there was a significant increase in the expression of caspase-3, heat shock protein 70, lipopolysaccharide-induced tumor necrosis factor-α, transcriptional enhanced associate domain and yorkie in intestinal and hepatopancreatic tissues. Following heat stress, the number of potentially beneficial bacteria (Rhodobacteraceae and Gemmonbacter) increased which maintain balance and promote vitamin synthesis. Intestinal transcriptome analysis revealed 852 differentially expressed genes in the heat stress group compared with the control group. KEGG functional annotation results showed that the endocrine system was the most abundant in Organismal systems followed by the immune system. These results indicated that heat stress leads to tissue damage in shrimp, however the shrimp may respond to stress through a coordinated interaction strategy of the endocrine system, immune system and gut microbiota. This study revealed the response mechanism of L. vannamei to acute heat stress and potentially provided a theoretical foundation for future research on shrimp environmental adaptations.

Toxic effects of glyphosate on histopathology and intestinal microflora of juvenile Litopenaeus vannamei.

Glyphosate is a widely used broad-spectrum herbicide, its pollution to the surrounding conditions can't be ignored. It has been reported that glyphosate has poisonous impacts on aquatic animals. In this study, juvenile Litopenaeus vannamei (L. vannamei) was exposed to glyphosate, and the lethal concentration 50 (LC50) of glyphosate to juvenile L. vannamei for 48 h was 47.6 mg/L. The histological analysis for intestine and hepatopancreas and the intestinal microorganisms of L. vannamei were evaluated after 48 h of exposure to glyphosate with LC50. The histological analysis results showed that the lumen of hepatic tubules was diffused and deformed, the hepatic tubules were ruptured and intestinal villi were fallen off seriously after exposure to glyphosate for 48 h Moreover, the intestinal microbial composition and structure of L. vannamei were changed, with the abundance of Alphaproteobacteria increased significantly. The abundance of Rhodobacteraceae, Vibrio and Legionella increased, but there was no significant difference. The abundance of Bacillus, Paraburkholderia, Enhydrobacter, Comamonas and Alkanindiges decreased significantly. However, the homeostasis of intestinal microorganisms was destroyed. Phenotypic prediction of the two groups of microorganisms revealed a significant increase in the abundance of Facultatively Anaerobic in the glyphosate challenged group. This study suggested that hepatopancreas and intestinal tissue of L. vannamei were seriously damaged after 48 h of exposure to glyphosate with LC50, and intestinal microbial homeostasis was disrupted.