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.

Characterization, Expression, and Functional Analysis of the Northern Snakehead (Channa argus) Hepcidin.

Hepcidin, an antimicrobial peptide (AMP), is a well-conserved molecule present in various species such as fish, amphibians, birds, reptiles, and mammals. It exhibits broad-spectrum antimicrobial activity and holds a significant role in the innate immune system of host organisms. The northern snakehead (Channa argus) has become a valuable freshwater fish in China and Asia. In this investigation, the cDNA encoding the hepcidin gene of northern snakehead was cloned and named caHep. The amino acid sequences and protein structure of caHep are similar to those of hepcidins from other fish. The eukaryotic expression product of the caHep gene showed broad-spectrum antibacterial activity. Scanning electron microscope analysis indicated that the caHep peptide inhibited bacterial growth by damaging their cell membranes. Lipopolysaccharide (LPS) injection induced significant expression of caHep, implying the involvement of caHep in the innate immune response of northern snakeheads. This investigation showed that the caHep peptide is potentially a robust antibacterial drug against bacterial diseases in aquaculture animals.

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.

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.

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.

The complete mitochondrial genome of Diagramma pictum (Perciformes: Haemulidae).

The painted sweetlips Diagramma pictum (Thunberg 1792) is an important fish for commercial fisheries which is widely distributed in the Indo-West Pacific Ocean. It can change its external coloration and pattern during their lives. The complete mitochondrial genome of D. pictum was determined in this study. The genome was 16,531 bp in length and consisted of 13 protein coding genes, 22 transfer RNA (tRNA), 2 ribosomal RNA (rRNA), and one noncoding control region. The overall base composition was estimated to be A: 27.5%; T: 24.7%; C: 30.9% and G: 16.9% with AT bias of 52.2%. The molecular phylogenetic result revealed that D. pictum did not form an independent branch but was tightly clustered inside the Plectorhinchus groups, closely related to the species Plectorhinchus chaetodonoides, indicating the close relationships between genera Diagramma and Plectorhinchus. These results may provide important genomic information for species evolution and mitogenome based phylogenetic analyses of D. pictum in the family Haemulidae.

[Role of enamel matrix proteins in inducing biomimetic mineralization of the enamel: a study with quartz crystal microbalance technique].

OBJECTIVE: To investigate the adsorption behavior of enamel matrix proteins (EMPs) on the enamel surface and study their effect on biomineralization of enamel using quartz crystal microbalance (QCM) technique. METHODS AND RESULTS: The EMPs were adsorbed on the enamel surface to form a protein film, which was soaked in simulated body fluid solutions. After 30 days of biomimetic mineralization, the hydroxyapatite nucleation, growth and aggregation occurred with hydroxyapatite crystal formation on the enamel surface. CONCLUSION: The EMPs play a key role in regulating enamel mineralization.