Protective effects of butyric acid during heat stress on the survival, immune response, histopathology, and gene expression in the hepatopancreas of juvenile pacific shrimp (L. Vannamei).

This study looked at the effects of adding butyric acid (BA) to the diets of juvenile Pacific shrimp and how it affected their response to survival, immunity, histopathological, and gene expression profiles under heat stress. The shrimp were divided into groups: a control group with no BA supplementation and groups with BA inclusion levels of 0.5 %, 1 %, 1.5 %, 2 %, and 2.5 %. Following the 8-week feeding trial period, the shrimp endured a heat stress test lasting 1 h at a temperature of 38 °C. The results showed that the control group had a lower survival rate than those given BA. Interestingly, no mortality was observed in the group receiving 1.5 % BA supplementation. Heat stress had a negative impact on the activities of alkaline phosphatase (AKP) and acid phosphatase (ACP) in the control group. Still, these activities were increased in shrimp fed the BA diet. Similar variations were observed in AST and ALT fluctuations among the different groups. The levels of triglycerides (TG) and cholesterol (CHO) increased with high temperatures but were reduced in shrimp-supplemented BA. The activity of an antioxidant enzyme superoxide dismutase (SOD) increased with higher BA levels (P < 0.05). Moreover, the groups supplemented with 1.5 % BA exhibited a significant reduction in malondialdehyde (MDA) content (P < 0.05), suggesting the potential antioxidant properties of BA. The histology of the shrimp's hepatopancreas showed improvements in the groups given BA. Conversely, the BA significantly down-regulated the HSPs and up-regulated MnSOD transcript level in response to heat stress. The measured parameters determine the essential dietary requirement of BA for shrimp. Based on the results, the optimal level of BA for survival, antioxidant function, and immunity for shrimp under heat stress is 1.5 %.

Implications on growth performance, glucose metabolism, PI3K/AKT pathway, intestinal flora induced by dietary taurine in a high-carbohydrate diet for grass carp (Ctenopharyngodon idella).

An 8-week experiment was performed to investigate the influence on growth performance, plasma biochemistry, glucose metabolism and the insulin pathway of supplementation of dietary taurine to a high-carbohydrate diet for grass carp. In this study, fish were fed diets at one of two carbohydrate levels, 31·49 % (positive control) or 38·61 % (T00). The high-carbohydrate basal diet (T00), without taurine, was supplemented with 0·05 % (T05), 0·10 % (T10), 0·15 % (T15) or 0·20 % (T20) taurine, resulting in six isonitrogenous (30·37 %) and isolipidic (2·37 %) experimental diets. The experimental results showed that optimal taurine level improved significantly weight gain, specific growth rate (SGR), feed utilisation, reduced plasma total cholesterol levels, TAG and promoted insulin-like growth factor level. Glucokinase, pyruvate kinase and phosphoenolpyruvate carboxykinase activities showed a quadratic function model with increasing dietary taurine level, while hexokinase, fatty acid synthetase activities exhibited a positive linear trend. Optimal taurine supplementation in high-carbohydrate diet upregulated insulin receptor (Ir), insulin receptor substrate (Irs1), phosphatidylinositol 3-kinase (pi3k), protein kinase B (akt1), glycogen synthase kinase 3 ß (gs3kß) mRNA level and downregulated insulin-like growth factor (igf-1), insulin-like growth factor 1 receptor (igf-1R) and Fork head transcription factor 1 (foxo1) mRNA level. The above results suggested that optimal taurine level could improve growth performance, hepatic capacity for glycolipid metabolism and insulin sensitivity, thus enhancing the utilisation of carbohydrates in grass carp. Based on SGR, dietary optimal tributyrin taurine supplementation in grass carp was estimated to be 0·08 %.

An integrated study of glutamine alleviates enteritis induced by glycinin in hybrid groupers using transcriptomics, proteomics and microRNA analyses.

Glutamine has been used to improve intestinal development and immunity in fish. We previously found that dietary glutamine enhances growth and alleviates enteritis in juvenile hybrid groupers (Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂). This study aimed to further reveal the protective role of glutamine on glycinin-induced enteritis by integrating transcriptome, proteome, and microRNA analyses. Three isonitrogenous and isolipidic trial diets were formulated: a diet containing 10% glycinin (11S group), 10% glycinin diet supplemented with 2% alanine-glutamine (Gln group), and a diet containing neither glycinin nor alanine-glutamine (fishmeal, FM group). Each experimental diet was fed to triplicate hybrid grouper groups for 8 weeks. The analysis of intestinal transcriptomic and proteomics revealed a total of 570 differentially expressed genes (DEGs) and 169 differentially expressed proteins (DEPs) in the 11S and FM comparison group. Similarly, a total of 626 DEGs and 165 DEPs were identified in the Gln and 11S comparison group. Integration of transcriptome and proteome showed that 117 DEGs showed consistent expression patterns at both the transcriptional and translational levels in the Gln and 11S comparison group. These DEGs showed significant enrichment in pathways associated with intestinal epithelial barrier function, such as extracellular matrix (ECM)-receptor interaction, tight junction, and cell adhesion molecules (P < 0.05). Further, the expression levels of genes (myosin-11, cortactin, tenascin, major histocompatibility complex class I and II) related to these pathways above were significantly upregulated at both the transcriptional and translational levels (P < 0.05). The microRNA results showed that the expression levels of miR-212 (target genes colla1 and colla2) and miR-18a-5p (target gene colla1) in fish fed Gln group were significantly lower compared to the 11S group fish (P < 0.05). In conclusion, ECM-receptor interaction, tight junction, and cell adhesion molecules pathways play a key role in glutamine alleviation of hybrid grouper enteritis induced by high-dose glycinin, in which miRNAs and target mRNAs/proteins participated cooperatively. Our findings provide valuable insights into the RNAs and protein profiles, contributing to a deeper understanding of the underlying mechanism for fish enteritis.

Integrated Metabolome and Transcriptome Analyses Reveal the Efficacy of Steroidal Saponins for Glucose and Lipid Metabolism in Hybrid Grouper (♀Epinephelus fuscoguttatus × â™‚Epinephelus lanceolatu) Fed Higher-Lipid Diets.

An analysis of the extent of the effect of steroidal saponin addition on glucose and lipid metabolism in hybrid grouper liver was performed at the transcriptomic and metabolomic levels. Feeds (52% crude protein, 14% crude lipid) were prepared containing 0% (S0), 0.1% (S0.1), and 0.2% (S0.2) steroidal saponins. After eight weeks of feeding trial, compared to the S0 group, the activities of serum albumin, alanine aminotransferase, and aspartate transaminase were significantly lower and the activities of lysozyme, acid phosphatase, and alkaline phosphatase were significantly higher in the S0.1 group (p < 0.05). The superoxide dismutase, catalase, and glutathione peroxidase activities in the livers of the S0.1 group were significantly higher than those of the S0 group, while the malondialdehyde content was significantly lower than that of the S0 group (p < 0.05). There were forty-two differentially expressed genes and thirty-two differential metabolites associated with glucose and lipid metabolism enriched using KEGG and GO. In the S0 group, the expression of prostaglandin-endoperoxide synthase 1, prostaglandin E synthase 1, and thromboxane-2 synthase mRNA was significantly higher than in the S0.1 group (p < 0.05). The expression levels of genes in the S0 group were significantly higher than those in the S0.1 group (p < 0.05), including for glycogen synthase kinase, glucose-6-phosphatase catalytic subunit 2, fructose-1,6-bisphosphatase, phosphoenolpyruvate carboxykinase, glucose transporter 4, and malate dehydrogenase. The expression of mRNA such as fatty acid synthase, acetyl-CoA carboxylase, and sterol regulatory element-binding protein 1 was significantly lower in the S0.1 group than in the S0 group, while the expression of carnitine acyltransferase 1, acyl-CoA synthetase, and acyl-CoA dehydrogenase genes was significantly higher in the S0 group (p < 0.05). In summary, glycogen synthesis, gluconeogenesis, and the arachidonic acid metabolism pathway were inhibited by 0.1% steroidal saponins, and glycogenolysis, glycolysis, the tricarboxylic acid cycle, and the fatty acid ß-oxidation pathway were activated. This study aims to provide a reference for the formulation of grouper feeds with a higher crude-lipid level.

Glutamine improves growth and intestinal health in juvenile hybrid groupers fed high-dose glycinin.

Glutamine addition can improve immunity and intestinal development in fish. This study examined the protective roles of glutamine on growth suppression and enteritis induced by glycinin in juvenile hybrid groupers (female Epinephelus fuscoguttatus × male Epinephelus lanceolatus). The experiment set four isonitrogenous and isolipidic trial diets: a diet containing 10% glycinin (11S), 10% of 11S diet supplemented with 1% or 2% alanine-glutamine (1% or 2% Ala-Gln), and a diet containing neither 11S nor Ala-Gln (FM). A feeding trial was conducted in hybrid grouper for 8 weeks. Weight gain and specific growth rates in Groups 1% and 2% Ala-Gln were significantly higher than those of the 11S group but were similar to those of the FM group. The intestinal muscular layer thickness, plica height and width of the 2% Ala-Gln group were significantly higher than those of Group 11S. The enterocyte proliferation efficiency of the 11S group was significantly lower compared to other groups. Compared with the 11S group, Groups 1% and 2% Ala-Gln fish had increased intestinal lysozyme activities, complement 3 and immunoglobulin M as well as cathelicidin contents. The mRNA levels of tnf-α, il-1ß, ifn-α, and hsp70 genes were more downregulated in Groups 1% and 2% Ala-Gln than in Group 11S. Compared with FM group, fish from the 11S group had significantly lower mRNA levels of myd88, ikkß, and nf-κb p65 genes. These three values in the 2% Ala-Gln group were significantly lower than those in Group 11S but not significantly different from those of Group FM. The relative abundance of Vibrio in Group 11S was higher than that in Groups FM and 2% Ala-Gln. Intestinal glutamine, glutaminase, glutamic acid, α-ketoglutarate, malate dehydrogenase and ATP contents were higher in Groups 1% and 2% Ala-Gln than in Group 11S. These results suggest that glutamine is a useful feed additive to enhance growth and intestinal immunity, alleviate inflammation, and modulate gut microbiota in hybrid grouper fed high-dose glycinin.

Scoparia dulcis L. Extract Relieved High Stocking Density-Induced Stress in Crucian Carp (Carassius auratus).

The objective of this study was to investigate the effects of Scoparia dulcis extract (SDE) on stress induced by high stocking density and Cu and trichlorfon exposure in crucian carp (Carassius auratus). The results showed that these stressors exerted detrimental effects in fish, such as inhibition of growth performance, reduced feed intake, and interruption of fish locomotion. Under high stocking density, dietary SDE supplementation increased the content of reduced glutathione (GSH) and the activities of amylase, catalase (CAT), and glutathione reductase (GR) and decreased the content of malonaldehyde (MDA) in the intestine of crucian carp. A similar trend was presented in the hepatopancreas under Cu exposure. Dietary SDE supplementation enhanced the activities of CAT, superoxide dismutase (SOD), glutathione peroxidase (GPx), lactate dehydrogenase, glutamate-oxaloacetate transaminase, and glutamate-pyruvate transaminase in the muscle of crucian carp under trichlorfon exposure. The optimum dietary SDE supplementation levels were 4.07, 4.33, and 3.95 g kg-1 diet based on the recovery rate of weight gain (RWG), feed intake (FI), and inhibitory rate of rollover (IR) for crucian carp under high stocking density and Cu and trichlorfon exposure, respectively. Overall, dietary supplementation with SDE may be a useful nutritional strategy for relieving these stresses in aquatic animals.

The Protective Effect of a Dietary Extract of Mulberry (Morus alba L.) Leaves against a High Stocking Density, Copper and Trichlorfon in Crucian Carp (Carassius auratus).

This study was designed to examine the protective effects of the extract of mulberry (Morus alba L.) leaves (EML) on crucian carp (Carassius auratus) against a high stocking density, Cu exposure and trichlorfon exposure, which adversely impact fish growth performance, feed intake and fish locomotion. High stocking densities decreased the activities of amylase, lipase, trypsin, Na+/K+-ATPase and alkaline phosphatase (AKP), and increased the content of malonaldehyde (MDA) in fish digestive organs, indicating an impairment of the digestive function and a disturbance of the antioxidant status. Cu exposure increased the activities of glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) in fish digestive organs, suggesting the activation of amino acid metabolism. Furthermore, trichlorfon exposure reduced the activities of lactate dehydrogenase (LDH), glutathione reductase (GR), GOT and GPT, and the capacities of the anti-superoxide anion (ASA) and anti-hydroxyl radical (AHR) in fish muscles, indicating a disruption of the bioenergetic homeostasis and antioxidant status. Our present study indicates that dietary EML supplementation relieved the detrimental effects induced by these stressors.

Generalized epidemic model incorporating non-Markovian infection processes and waning immunity.

The Markovian approach, which assumes exponentially distributed interinfection times, is dominant in epidemic modeling. However, this assumption is unrealistic as an individual's infectiousness depends on its viral load and varies over time. In this paper, we present a Susceptible-Infected-Recovered-Vaccinated-Susceptible epidemic model incorporating non-Markovian infection processes. The model can be easily adapted to accurately capture the generation time distributions of emerging infectious diseases, which is essential for accurate epidemic prediction. We observe noticeable variations in the transient behavior under different infectiousness profiles and the same basic reproduction number R_{0}. The theoretical analyses show that only R_{0} and the mean immunity period of the vaccinated individuals have an impact on the critical vaccination rate needed to achieve herd immunity. A vaccination level at the critical vaccination rate can ensure a very low incidence among the population in the case of future epidemics, regardless of the infectiousness profiles.

Host gut-derived Bacillus probiotics supplementation improves growth performance, serum and liver immunity, gut health, and resistive capacity against Vibrio harveyi infection in hybrid grouper (♀Epinephelus fuscoguttatus × â™‚Epinephelus lanceolatus).

Several reports have revealed the vital role that probiotics play in fish growth and health. However, few works are available for host gut-derived probiotics on the growth, immunity, and gut microbiota of fish, especially in hybrid grouper (♀Epinephelus fuscoguttatus × â™‚Epinephelus lanceolatus) due to their isolation difficulty and functional verification. This study aimed at assessing 3 host gut-derived Bacillus species' effects on the growth, immune and antioxidant-biochemical responses, haematological parameters, intestinal morphology, immune-related gene expression, gut microbiota, and disease resistance against Vibrio harveyi in hybrid grouper. A total of 480 hybrid grouper (initial weight = 9.03 ± 0.02 g) were randomly allotted into 4 groups, namely, the group fed a basal diet without probiotic inclusion (control, B0), the group fed the basal diet with Bacillus velezensis GPSAK4 (BV), the group fed the basal diet with Bacillus subtilis GPSAK9 (BS), and the group fed the basal diet with Bacillus tequilensis GPSAK2 (BT) strains at 1.0 × 109 CFU/g. After a 6-week feeding trial, the results revealed significant improvements (P < 0.05) in the growth performance, whole fish-body proximate composition, blood haematological parameters, serum, liver, and intestinal biochemical indexes, intestinal morphology, and protection against V. harveyi pathogen in the probiotic-treated groups compared with the untreated. Additionally, the expressions of intestinal tight junction genes (occludin and ZO1), pro- and anti-inflammatory genes, including IL1ß, IL6, IL8, TNFα, MyD88, IL10, and TGFß, were upregulated (P < 0.05) after Bacillus species administration. Host gut-derived Bacillus supplementation shaped the gut microbiota by significantly increasing (P < 0.05) the relative abundance of Proteobacteria, Bacteroidetes, Actinobacteria (except the BS group), Acidobacteria (except the BT group), Cyanobacteria (except the BV and BT groups), and Verrucomicrobia phyla, as well as known beneficial genera (Romboutsia, Turicibacter, Epulopiscium, Clostridium_sensu_stricto 1 and 13, Lactobacillus, and Bacillus), but significantly decreased (P < 0.05) the abundance of Firmicutes, Chloroflexi, and Fusobacteria phyla, and purported pathogenic genera (Staphylococcus and Photobacterium) compared with the control group. Collectively, the results suggest that B. velezensis GPSAK4, B. subtilis GPSAK9 (especially this strain), B. tequilensis GPSAK2 dietary supplementation at 1.0 × 109 CFU/g has positive effects on the intestinal health of hybrid grouper via microbial composition modulation, thus enhancing the assimilation and absorption of nutrients to boost fish growth, immunity, and disease resistance.

Effects of Clostridium butyricum on Growth Performance, Intestinal Health, and Disease Resistance of Hybrid Grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) Fed with Cottonseed Protein Concentrate (CPC) Replacement of Fishmeal.

An 8-week feeding trial was conducted to investigate the effects of C. butyricum on the growth performance, microbiota, immunity response, and disease resistance in hybrid grouper fed with cottonseed protein concentrate (CPC) replacement of fishmeal. Six groups of isonitrogenous and isolipid diets were formulated including a positive control group (50% fishmeal, PC), a negative control group (CPC replaced 50% of fishmeal protein, NC), and Clostridium butyricum supplemented with 0.05% (C1, 5 × 108 CFU/kg), 0.2% (C2, 2 × 109 CFU/kg), 0.8% (C3, 8 × 109 CFU/kg), and 3.2% (C4, 3.2 × 1010 CFU/kg), respectively, to the NC group. The results showed that weight gain rate and specific growth rate were significantly higher in the C4 group than that in the NC group (P < 0.05). After supplementation with C. butyricum, the amylase, lipase, and trypsin activities were significantly higher than the NC group (P < 0.05; except group C1), and the same results were obtained for intestinal morphometry. The intestinal proinflammatory factors were significantly downregulated, and the anti-inflammatory factors were significantly upregulated in the C3 and C4 groups compared with the NC group after supplementation with 0.8%-3.2% C. butyricum (P < 0.05). At the phylum level, the PC, NC, and C4 groups were dominated by the Firmicutes and the Proteobacteria. At the genus level, the relative abundance of Bacillus in the NC group was lower than that in the PC and C4 groups. After supplementation with C. butyricum, grouper in the C4 group showed significantly higher resistance to V. harveyi than the NC group (P < 0.05). Above all, taking into account the effects of immunity and disease resistance, it was recommended to supplement 3.2% C. butyricum in the diet of grouper fed the replacement of 50% fishmeal protein by CPC.

Effects of partially replacing fishmeal with corn gluten meal on growth, feed utilization, digestive enzyme activity, and apparent nutrient digestibility for juvenile white shrimp, Litopenaeus vannamei.

This experiment was conducted to assess the possibility of replacing fishmeal (FM, Fishmeal content of the control group: 30%) with corn gluten meal (CGM) at the following levels: 0, 10, 20, 30, 40, and 60%. The experimental diets, formulated to be isonitrogenous and isocaloric, were studied for their effects on growth, feed utilization, digestive enzyme activity and apparent nutrient digestibility in juvenile white shrimps, Litopenaeus vannamei (initial mean weight = 0.71 ± 0.01 g). Seven hundred twenty healthy and uniformed-size shrimp were distributed to six groups of three replicates, each with 40 shrimp in each tank (0.5 m3). Each experimental diet was fed to shrimp four times daily to apparent satiation at 7:00, 11:30, 17:00, and 21:30, respectively, for 8 weeks. At the end of the experiment, the total weight of fish in each tank was weighed and randomly selected for testing, including fish nutrient composition and digestive enzyme activity. Results showed that no significant differences were observed in the weight gain rate (WGR), feed coefficient rate (FCR) and specific growth rate (SGR) of shrimp after 30% FM was replaced with CGM (P > 0.05), but these indicators significantly decreased at higher replacement rates. As CGM content increased, the content of crude protein and phosphorus in the shrimp decreased significantly (P < 0.05), whereas the crude fat content first increased significantly and then decreased (P < 0.05). Compared to the control group, the protease activity was significantly lower in the 40% group and the lipase activity was significantly lower in the 60% group (P < 0.05). Amylase activity was significantly increased with increasing CGM levels (P < 0.05). The digestibility of protein and lipid was significantly reduced by CGM replacement of more than 30% FM (P < 0.05). As CGM content increased, the digestion of energy and dry matter was first significantly increased and then significantly decreased (P < 0.05). In the 30, 40, and 60% groups, the digestibility of all amino acids (AA), except methionine (Met), arginine (Arg) and serine (Ser), was significantly lower than in the control group (P < 0.05). In summary, FM could be partially replaced by CGM in the feed of L. vannamei. Based on the broken-line regression analysis of WGR, the optimal dietary CGM replacement was 27.47%.

The Effect of Glycerol Monolaurate on Intestinal Health and Disease Resistance in Cage-Farmed Juvenile Pompano Trachinotus ovatus.

This research studied the effects of glycerol monolaurate (GML) to diets on the digestive capacity, intestinal structure, intestinal microbiota, and disease resistance for juvenile pompano Trachinotus ovatus (mean weight = 14.00 ± 0.70 g). T. ovatus were, respectively, fed six diets containing 0.00, 0.05, 0.10, 0.15, 0.20, and 0.25% GML for 56 days. The highest weight gain rate was observed in the 0.15% GML group. In the intestine, amylase activities in the 0.10, 0.15, 0.20, and 0.25% GML groups were significantly increased, compared with 0.00% GML group (P < 0.05). Lipase activities in the 0.10 and 0.15% GML groups were significantly increased (P < 0.05). Similar significant elevations in the protease activities were also found in the 0.10, 0.15, and 0.20% GML groups (P < 0.05). Amylase activities were significantly higher in the 0.10, 0.15, 0.20, and 0.25% GML groups than that in the 0.00% GML group (P < 0.05). Villus lengths (VL) and muscle thicknesses (MT) of the 0.05, 0.10, 0.15, and 0.20% GML groups were significantly enhanced, and the villus widths (VW) in the 0.05, 0.10, and 0.15% groups were significantly increased (P < 0.05). Additionally, 0.15% GML significantly improved the intestinal immunity by upregulating interleukin 10 (il-10), increasing beneficial bacteria abundances (e.g., Vibrio, Pseudomonas, and Cetobacterium), downregulating nuclear factor kappa b (nf-κb) and interleukin 8 (il-8), and decreasing harmful bacteria abundances (e.g., Brevinema and Acinetobacter) (P < 0.05). After challenge test, GML significantly increased the survival rate (80%-96%) (P < 0.05). In addition, ACP and AKP activities in the GML-supplemented groups were significantly higher than those in the 0.00% GML group, and LZM activity was significantly higher in the 0.05, 0.10, 0.15, and 0.20% GML groups than that in the 0.00% GML group (P < 0.05). In summary, 0.15% GML significantly promoted the intestinal digestibility, improved the intestinal microflora, regulated intestinal immune-related genes, and increased resistance to V. parahaemolyticus of juvenile pompano T. ovatus.

The isolation, identification, whole-genome sequencing of Clostridium butyricum LV1 and its effects on growth performance, immune response, and disease-resistance of Litopenaeus vannamei.

In this study, a strain of Clostridium butyricum was isolated from the intestine of Litopenaeus vannamei with the method of anaerobic microbial isolation and culture. Next, the probiotic properties of LV1 were evaluated with susceptibility tests, tolerance tests, and whole genome sequencing in vivo and in vitro, followed by the analysis of the effect of LV1 on the growth performance, immune response, and disease resistance of Litopenaeus vannamei. According to the results, the 16 S rDNA sequence of LV1 was 100% homolofgous to the reference sequence of Clostridium butyricum. Moreover, LV1 was resistant to several antibiotics including amikacin, streptomycin, and gentamicin and highly tolerated artificial gastric and artificial intestinal fluids. The whole genome of LV1 was 4625,068 bp in size and included 4336 coding genes. Among these genes, GO, KEGG, and COG databases exhibited the highest number of genes annotated to metabolic pathway classes and 105 genes annotated as glycoside hydrolases. Meanwhile, 176 virulence genes were predicted. The use of diets supplemented with 1.2 × 109 CFU/kg of LV1 live cells significantly increased the weight gain and specific growth rates of Litopenaeus vannamei and the activity of serum superoxide dismutase, glutathione peroxidase, acid phosphatase, and alkaline phosphatase (P < 0.05). Meanwhile, the use of these diets markedly improved the relative expression of intestinal immunity- and growth-related genes. In conclusion, LV1 has excellent probiotic properties. Specifically, the addition of 1.2 × 109 CFU/kg of LV1 live cells to the diet improved the growth performance, immune response, and disease-resistance of Litopenaeus vannamei.

Preparation, Identification and Preliminary Application of the Fenvalerate Monoclonal Antibody in Six Kinds of Dark Tea.

Fenvalerate has the advantages of a broad insecticidal spectrum, high efficiency, low toxicity and low cost, and it is widely used in agriculture, especially in tea, resulting in the accumulation of fenvalerate residues in tea and the environment, posing a serious threat to human health. Therefore, the timely monitoring of fenvalerate residue dynamics is vital for ensuring the health of humans and the ecological environment, and it is necessary for establishing a fast, reliable, accurate and on-site method for detecting fenvalerate residues. Based on the methods of immunology, biochemistry and molecular biology, mammalian spleen cells, myeloma cells and mice were used as experimental materials to establish a rapid detection method of an enzyme-linked immunosorbent assay to detect the residues of fenvalerate in dark tea. Three cell lines-1B6, 2A11 and 5G2-that can stably secrete fenvalerate antibodies were obtained by McAb technology, and their sensitivities (IC50) were 36.6 ng/mL, 24.3 ng/mL and 21.7 ng/mL, respectively. The cross-reaction rates of the pyrethroid structural analogs were all below 0.6%. Six dark teas were used to detect the practical application of fenvalerate monoclonal antibodies. The sensitivity IC50 of the anti-fenvalerate McAb in PBS with 30% methanol is 29.12 ng/mL. Furthermore, a latex microsphere immunochromatographic test strip with an LOD of 10.0 ng/mL and an LDR of 18.9-357 ng/mL was preliminarily developed. A specific and sensitive monoclonal antibody for fenvalerate was successfully prepared and applied to detect fenvalerate in dark teas (Pu'er tea, Liupao tea, Fu Brick tea, Qingzhuan tea, Enshi dark tea and selenium-enriched Enshi dark tea). A latex microsphere immunochromatographic test strip was developed for the preparation of rapid detection test strips of fenvalerate.

Effects of Dietary Steroid Saponins on Growth Performance, Serum and Liver Glucose, Lipid Metabolism and Immune Molecules of Hybrid Groupers (♀Epinephelus fuscoguttatus × â™‚Epinephelus lanceolatu) Fed High-Lipid Diets.

High-lipid diets are attributed to excessive lipid deposition and metabolic disturbances in fish. The aim of this experiment was to investigate the effects of steroidal saponins on growth performance, immune molecules and metabolism of glucose and lipids in hybrid groupers (initial weight 22.71 ± 0.12 g) fed high-lipid diets. steroidal saponins (0%, 0.1% and 0.2%) were added to the basal diet (crude lipid, 14%) to produce three experimental diets, designated S0, S0.1 and S0.2, respectively. After an 8-week feeding trial, no significant differences were found between the S0 and S0.1 groups in percent weight gain, specific growth rate, feed conversion ratio, protein efficiency ratio and protein deposition rate (p > 0.05). All those in the S0.2 group were significantly decreased (p < 0.05). Compared to the S0 group, fish in the S0.1 group had lower contents of serum triglyceride and low-density lipoprotein cholesterol and higher high-density lipoprotein cholesterol and glucose (p < 0.05). The activities of superoxide dismutase, catalase and glutathione peroxidase were significantly higher, and malondialdehyde contents were significantly lower in the S0.1 group than in the S0 group (p < 0.05). Hepatic triglyceride, total cholesterol and glycogen were significantly lower in the S0.1 group than in the S0 group (p < 0.05). Activities of lipoprotein lipase, total lipase, glucokinase and pyruvate kinase, and gene expression of lipoprotein lipase, triglyceride lipase and glucokinase, were significantly higher in the S0.1 group than in the S0 group. Interleukin-10 mRNA expression in the S0.1 group was significantly higher than that in the S0 group, while the expression of interleukin-6 and tumor necrosis factor-α genes were significantly lower than those in the S0 group. In summary, adding 0.1% steroidal saponins to a high-lipid diet not only promoted lipolysis in fish livers, but also activated glycolysis pathways, thus enhancing the utilization of the dietary energy of the groupers, as well as supporting the fish's nonspecial immune-defense mechanism.

Effects of Carbohydrase Supplementation on Growth Performance, Intestinal Digestive Enzymes and Flora, Glucose Metabolism Enzymes, and glut2 Gene Expression of Hybrid Grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂) Fed Different CHO/L Ratio Diets.

An optimal carbohydrate-to-lipid (CHO: L) ratio facilitates fish growth and protein conservation, and carbohydrase promotes nutrient absorption. Therefore, an 8-week feeding trial was conducted to investigate the effects of carbohydrase supplementation on growth performance, intestinal digestive enzymes and flora, glucose metabolism enzymes and glut2 gene expression in juvenile hybrid grouper (Epinephelus fuscoguttatusâ™€× Epinephelus lanceolatus♂) fed different CHO: L ratios diets. L, M, and H represent CHO:L ratios of 0.91, 1.92 and 3.91, respectively. LE, ME, and HE represent CHO:L ratios of 0.91, 1.92, 3.91, respectively, supplemented with the same ratio of carbohydrase. Results showed that weight gain rate (WGR) and specific growth rate (SGR) reached a maximum in group M and were significantly enhanced by carbohydrase (p < 0.05). Crude lipid content decreased significantly with an increase in the dietary CHO:L ratio (p < 0.05). Significant increases in the trypsin (TRY) and amylase (AMS) activities and significant decreases in the lipase (LPS) activity were observed with increasing dietary CHO:L ratio, and the former two were significantly promoted by carbohydrase (p < 0.05). The content of liver and muscle glycogen increased significantly with the increasing dietary CHO:L ratio but decreased significantly after carbohydrase supplementation (p < 0.05). The glucokinase (GK), pyruvate kinase (PK), Phosphate 6 fructokinase-1 (PFK-1) and phosphoenolpyruvate kinase (PEPCK) activities increased significantly with increasing dietary CHO:L ratio (p < 0.05). Glut2 mRNA expression decreased significantly in liver and increased significantly in intestine with increasing dietary CHO:L ratio (p < 0.05). By linear discriminant analysis (LDA), the abundance of Alistipe was significantly higher in Group ME than in Group M. These results suggested that hybrid grouper can only moderately utilize dietary carbohydrate and lipid in diet, and a certain amount of high glycemic lipids occurred when fed with high-carbohydrate diets. By the weight gain for basis, the supplementation of carbohydrase in Group H with amylase, glycosylase, and pullulanase in a 1:1:1 ratio effectively lowered glycemic lipids, promoted the growth of grouper, digestive enzymes activities and carbohydrate metabolic enzyme, and glut2 gene expression in intestine, effectively balancing the negative effects of high-carbohydrate diet and improving the utilization of carbohydrate.

Soybean protein concentrate causes enteritis in juvenile pearl gentian groupers (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂).

Due to diminishing fish meal (FM) supplies, superb protein (PRO) sources are needed for aquaculture, such as soy-based PRO. However, these can cause enteritis and even intestinal injury in fish when used at high proportions in feed. This research examines the effects of substituting soybean protein concentrate (SPC) for FM on the growth performance and intestinal balance of pearl gentian groupers and investigates the mechanism of SPC-induced enteritis. Experimental fish (n = 720) were fed 1 of 3 following diets: (1) a 50% FM diet (control), (2) a diet with 20% of the FM substituted with SPC (group SPC20), and (3) a 40% SPC-substituted diet (SPC40). Fish were fed for 10 wk iso-nitrogenous (50% PRO) and iso-lipidic (10% lipid) diets. Groups SPC20 and SPC40 showed significantly lower developmental performance and intestinal structures than control. Group SPC40 had significantly higher expressions of pro-inflammatory-related genes, such as interleukin 1ß (IL1ß), IL12, IL17 and tumor necrosis factor α and significantly lower expressions of anti-inflammatory-related genes, such as IL5, IL10 and transforming growth factor ß1. Biochemical and 16S high-throughput sequencing showed that the abundance and functions of intestinal flora in group SPC40 were significantly affected (P < 0.05), and there were significant correlations between operational taxonomic unit abundance variations and inflammatory gene expressions at genus level (P < 0.05). The second- and third-generation full-length transcriptome sequence was used to analyze the mechanism of SPC-induced enteritis in pearl gentian groupers, which showed that enteritis induced by SPC may be caused by disturbances to intestinal immune function induced by an imbalance in intestinal nutrition and metabolism, such as the intestinal immunity network for IgA production pathway. However, it remains unclear as to which intestinal immune or nutritional imbalance is most important in enteritis development. This study provides a basis for further research into soy PRO-related enteritis in fish.

Effect of Rice Protein Meal Replacement of Fish Meal on Growth, Anti-Oxidation Capacity, and Non-Specific Immunity for Juvenile Shrimp Litopenaeus vannamei.

This study assessed the effect of rice protein meal replacement for fish meal on the growth, nonspecific immunity, and disease resistance on juvenile shrimp Litopenaeus vannamei. Six groups of iso-nitrogenous and iso-lipid feeds named FM, R10, R20, R40, R60, and R80 were prepared by replacing 0%, 10%, 20%, 40%, 60%, and 80% in FM protein with RPM, respectively, and then fed to the shrimps (0.54 ± 0.01 g). An amount of 720 healthy and evenly sized shrimps were allocated to six groups (three replicates per group) and fed four times a day (7:00, 11:00, 17:00 and 21:00) for eight weeks. Results revealed no significant differences in WG, FCR, and SGR of shrimps after replacing FM with 10% RPM (p > 0.05). In the R10 and R20 groups, SOD and T-AOC activities were significantly higher than those in the FM group, whereas the opposite was observed for MDA content (p < 0.05). CAT, ACP, and LZM were all significantly higher in the R10, R20, and R40 groups than in the FM group (p < 0.05). GSH-Px activity in the R10 group was significantly higher than the activity in the FM group (p < 0.05). AKP, PO, TYS, GPT, and GOT activities were significantly higher in the R10 group than in the FM group (p < 0.05). Compared to the FM group, the eukaryotic translation initiation factor 3K (eif3k) gene was significantly up-regulated in the R10 group, whereas the penaiedin 3a (pen 3a) and anti-lipopolysaccharide factor (alf) genes were significantly up-regulated in the R10 and R20 groups (p < 0.05). The crustin a (cru a), immune deficiency (imd), and lysozyme (lzm) mRNA levels were significantly higher in the R10, R20, and R40 groups than in the other groups (p < 0.05). The prophenoloxidase (PO) mRNA levels in the R20 group were significantly higher than those in the FM group (p < 0.05). The replacement of 10−40% of FM with RPM improved the gut flora composition of shrimps, increasing beneficial bacteria (Bacteroidetes) abundance and reducing harmful bacteria (Aspergillus and Vibrio) abundance. After the challenge test of Vibrio parahaemolyticus (7 days), the cumulative mortality in the R10 group significantly decreased (p < 0.05). In conclusion, replacement of 10% FM by RPM significantly improved digestibility, protein synthesis, antioxidant capacity, and disease resistance in L. vannamei.

Glycerol Monolaurate Alleviates Oxidative Stress and Intestinal Flora Imbalance Caused by Salinity Changes for Juvenile Grouper.

Groupers with an initial body weight of 9.10 ± 0.03 g were selected to investigate whether dietary addition of 0 (G0) and 1800 mg/kg glycerol monolaurate (GML, G1800) could alleviate the oxidative stress response and intestinal flora imbalance after 0, 6, 12, and 24 h of salinity change in grouper. Experimental results show that the dietary addition of GML significantly reduced the liver MDA content and increased the SOD activity of grouper. The gene expression of CAT and SOD increased and then decreased with time after adding 1800 mg/kg GML, and the highest values were significantly higher than those of the control group. Salinity change had a slight effect on the top four intestinal flora composition of grouper at 0, 12, and 24 h, with changes occurring only at 6 h when Cyanobacteria replaced Actinobacteria. The addition of dietary GML slowed down the intestinal flora disorder, inhibited the colonization of harmful bacterium Vibrio, and promoted the abundance of beneficial bacterium Bacillus. In conclusion, dietary GML significantly reduced the oxidative damage caused by sudden changes in salinity, improved the antioxidant capacity, and alleviated the intestinal flora imbalance in juvenile grouper.