-
Fish & Shellfish Immunology Jun 2024Excess utilization of plant protein sources in animal feed has been found to adversely affect the antioxidant properties and immunity of animals. While the role of gut...
Excess utilization of plant protein sources in animal feed has been found to adversely affect the antioxidant properties and immunity of animals. While the role of gut microbes in plant protein-induced inflammation has been identified in various models, the specific mechanisms regulating gut microbes in crustaceans remain unclear. Accordingly, this study was designed to investigate the effects of replacing fishmeal with soybean meal (SM) on the hepatopancreas antioxidant and immune capacities, and gut microbial functions of crayfish, as well as the potential microbial regulatory mechanisms. 750 crayfish (4.00 g) were randomly divided into five groups: SS0, SS25, SS50, SS75, and SS100, and fed diets with different levels of soybean meal substituted for fishmeal for six weeks. High SM supplementation proved detrimental to maintaining hepatopancreas health, as indicated by an increase in hemolymph MDA content, GPT, and GOT activities, the observed rupture of hepatopancreas cell basement membranes, along with the decreased number of hepatopancreatic F cells. Moreover, crayfish subjected to high SM diets experienced obvious inflammation in hepatopancreas, together with up-regulated mRNA expression levels of nfkb, alf, and tlr (p<0.05), whereas the lzm mRNA expression level exhibited the highest value in the SS25 group. Furthermore, hepatopancreas antioxidant properties highly attenuated by the level of dietary SM substitution levels, as evidenced by the observed increase in MDA content (p<0.05), decrease in GSH content (p<0.05), and inhabitation of SOD, CAT, GPx, and GST activities (p<0.05), along with down-regulated hepatopancreas cat, gpx, gst, and mmnsod mRNA expression levels via inhibiting nrf2/keap1 pathway. Functional genes contributing to metabolism identified that high SM diets feeding significantly activated lipopolysaccharide biosynthesis, revealing gut dysfunction acted as the cause of inflammation. The global microbial co-occurrence network further indicated that the microbes contributing more to serum indicators and immunity were in module eigengene 17 (ME17). A structural equation model revealed that the genes related to alf directly drove the serum enzyme activities through microbes in ME17, with OTU399 and OTU533 identified as major biomarkers and classified into Proteobacteria that secrete endotoxins. To conclude, SM could replace 25 % of fishmeal in crayfish diets without negatively affecting immunity, and antioxidant capacity. Excessive SM levels contributed to gut dysfunction and weakened the innate immune system of crayfish.
Topics: Animals; Astacoidea; Animal Feed; Glycine max; Antioxidants; Diet; Gastrointestinal Microbiome; Hepatopancreas; Immunity, Innate; Random Allocation; Intestines; Dietary Supplements
PubMed: 38701991
DOI: 10.1016/j.fsi.2024.109600 -
Chemosphere Feb 2024Selenium (Se), an indispensable micronutrient for living organisms, has been extensively studied for its heavy metal-detoxifying properties in diverse biological systems...
Selenium (Se), an indispensable micronutrient for living organisms, has been extensively studied for its heavy metal-detoxifying properties in diverse biological systems and tissues. Nevertheless, it is not entirely certain whether Se can effectively protect against Cadmium (Cd)-induced gut inflammation, especially in aquatic animals. In this study, we employed various approaches, including transcriptome profiling, histological examinations, assessment of antioxidant enzyme activities, and analysis of gut microbiota composition to investigate the effects on crayfish growth and intestinal health after exposure to dietary Cd (15 mg kg diet) and Se (15 mg kg diet) individually or in combination for 8 weeks. The results revealed that dietary Cd exposure resulted in reduced body weight and survival rates, along with an increased occurrence of intestinal inflammation. Nevertheless, Se supplementation proved effective in mitigating the adverse effects of Cd on growth and gut health. Se exhibited a remarkable ability to counteract the disruption of gut antioxidant abilities induced by dietary Cd, as evidenced by the observed increases in ROS and MDA contents, decrease in GSH levels, and inhibition of antioxidative enzyme activities. At the concentration of 6 mg kg in the diet, Se was found beneficial for maintaining gut microbiota richness and diversity. Among them, Flavobacterium, Thermomonas, and Chloronema displayed a weak negative correlation with the rate of gut inflammation. Meanwhile, the levels of short chain fatty acids (SCFAs), including acetic acid (AA) and butanoic acid (BA), showed a significant increase in the Se-Cd group compared to the Cd-only group. Furthermore, transcriptome analysis exhibited significant responses of the PI3K/Akt and NF-κB pathways following crayfish exposure to dietary Se and Cd, either separately or in combination. In short, this study provides a new evidence regarding the molecular mechanisms through which Se could regulate the PI3K/Akt and NF-κB pathways, either directly or indirectly via ROS and SCFAs, thereby alleviating Cd-induced gut inflammation in crayfish.
Topics: Animals; Selenium; Antioxidants; Cadmium; NF-kappa B; Astacoidea; Proto-Oncogene Proteins c-akt; Phosphatidylinositol 3-Kinases; Reactive Oxygen Species; Inflammation
PubMed: 38040256
DOI: 10.1016/j.chemosphere.2023.140814 -
PloS One 2024Although behavioural defensive responses have been recorded several times in both laboratory and natural habitats, their neural mechanisms have seldom been investigated....
Although behavioural defensive responses have been recorded several times in both laboratory and natural habitats, their neural mechanisms have seldom been investigated. To explore how chemical, water-borne cues are conveyed to the forebrain and instruct behavioural responses in anuran larvae, we conditioned newly hatched agile frog tadpoles using predator olfactory cues, specifically either native odonate larvae or alien crayfish kairomones. We expected chronic treatments to influence the basal neuronal activity of the tadpoles' mitral cells and alter their sensory neuronal connections, thereby impacting information processing. Subsequently, these neurons were acutely perfused, and their responses were compared with the defensive behaviour of tadpoles previously conditioned and exposed to the same cues. Tadpoles conditioned with odonate cues differed in both passive and active cell properties compared to those exposed to water (controls) or crayfish cues. The observed upregulation of membrane conductance and increase in both the number of active synapses and receptor density at the postsynaptic site are believed to have enhanced their responsiveness to external stimuli. Odonate cues also affected the resting membrane potential and firing rate of mitral cells during electrophysiological patch-clamp recordings, suggesting a rearrangement of the repertoire of voltage-dependent conductances expressed in cell membranes. These recorded neural changes may modulate the induction of an action potential and transmission of information. Furthermore, the recording of neural activity indicated that the lack of defensive responses towards non-native predators is due to the non-recognition of their olfactory cues.
Topics: Animals; Larva; Predatory Behavior; Cues; Anura; Olfactory Receptor Neurons; Astacoidea
PubMed: 38696517
DOI: 10.1371/journal.pone.0302728 -
Comparative Biochemistry and... Dec 2023Sexual dimorphism widely exists in crustaceans. However, sex differences in the hypoxia tolerance of crayfish have rarely been reported. In this study, the differences...
Sexual dimorphism widely exists in crustaceans. However, sex differences in the hypoxia tolerance of crayfish have rarely been reported. In this study, the differences in hypoxia tolerance between the two sexes of crayfish were assessed according to mortality, pathological features of hepatopancreas, antioxidant enzyme activity and differentially expressed genes (DEGs) analysis using transcriptome. The results showed that male crayfish displayed significantly higher mortality than the female under hypoxia stress (p < 0.05). Furthermore, female crayfish demonstrated higher levels of antioxidant enzyme activity. Hematoxylin-eosin staining analysis revealed that the damage of hepatopancreas was more severe in the male crayfish compared to the female crayfish. Additionally, there was higher expression level of the DEGs in hypoxia-inducible factor (HIF) pathway and higher energy metabolism level in the female compared to the male. Together, these findings suggest that the female crayfish with higher antioxidant ability and energy metabolism level exhibits stronger hypoxia tolerance than the male crayfish, providing the theoretical support for investigating sex differences in hypoxia tolerance among crustaceans.
Topics: Female; Male; Animals; Antioxidants; Astacoidea; Sex Characteristics; Energy Metabolism; Hypoxia
PubMed: 37683360
DOI: 10.1016/j.cbd.2023.101136 -
Fish & Shellfish Immunology May 2024Serine protease inhibitors Kazal type (SPINKs) function in physiological and immunological processes across multicellular organisms. In the present study, we identified...
Serine protease inhibitors Kazal type (SPINKs) function in physiological and immunological processes across multicellular organisms. In the present study, we identified a SPINK gene, designated as CqSPINK, in the red claw crayfish Cherax quadricarinatus, which is the ortholog of human SPINK5. The deduced CqSPINK contains two Kazal domains consisting of 45 amino acid residues with a typical signature motif C-X-C-X-PVCG-X-Y-X-C-X-C-X-C. Each Kazal domain contains six conserved cysteine residues forming three pairs of disulfide bonds, segmenting the structure into three rings. Phylogenetic analysis revealed CqSPINK as a homolog of human SPINK5. CqSPINK expression was detected exclusively in hepatopancreas and epithelium, with rapid up-regulation in hepatopancreas upon Vibrio parahaemolyticus E1 challenge. Recombinant CqSPINK protein (rCqSPINK) was heterologously expressed in Escherichia coli and purified for further study. Proteinase inhibition assays demonstrated that rCqSPINK could potently inhibit proteinase K and subtilisin A, weakly inhibit α-chymotrypsin and elastase, but extremely weak inhibit trypsin. Furthermore, CqSPINK inhibited bacterial secretory proteinase activity from Bacillus subtilis, E. coli, and Staphylococcus aureus, and inhibited B. subtilis growth. These findings suggest CqSPINK's involvement in antibacterial immunity through direct inhibition of bacterial proteases, contributing to resistance against pathogen invasion.
Topics: Humans; Animals; Serine Proteinase Inhibitors; Astacoidea; Phylogeny; Escherichia coli; Recombinant Proteins; Bacteria
PubMed: 38537926
DOI: 10.1016/j.fsi.2024.109525 -
The Science of the Total Environment Dec 2023Integrated rice-crayfish farming is an ecological rice farming mode. However, limited research has examined the comprehensive impacts of greenhouse gas (GHG) emissions,...
Integrated rice-crayfish farming is an ecological rice farming mode. However, limited research has examined the comprehensive impacts of greenhouse gas (GHG) emissions, nitrogen (N) uptake, and N utilization in rice under this farming modality. Herein, a dual-factor experiment was performed from 2021 to 2022 to assess the comprehensive impacts of N application and rice farming mode on greenhouse gas (GHG) emissions, N uptake, N utilization, and rice yield in paddy fields. Under N application, the rice-crayfish co-culture exhibits a 2.3 % decrease in global warming potential (GWP) and a 17.3 % increase in greenhouse gas intensity relative to the rice monoculture. Moreover, the N uptake of rice within the rice-crayfish co-culture is 5.2 %-10.4 % higher than that in the rice monoculture. However, owing to low rice yield under the rice-crayfish co-culture, its N partial factor productivity decreases by 5.6 %-22.6 %, while N agronomic efficiency is reduced by 18.3 %-46.9 % compared with the rice monoculture. In addition, N application significantly inhibits CH emissions from paddy fields in the rice-crayfish co-culture mode. Compared with no N application, the CH emissions and GWP of N-applied treatment are decreased by 12.1 %-31.0 % and 6.0 %-15.8 %, respectively. Hence, N regulation might reduce GHG emissions in rice-aquatic animal co-culturing agriculture. Collectively, the results of this study suggest that switching from a rice monoculture to rice-crayfish co-culture can mitigate GHG emissions and promote rice N uptake; however, continuously improving the productivity of co-culturing agriculture is key to achieving high N utilization efficiency and low environmental impact.
Topics: Animals; Greenhouse Gases; Oryza; Astacoidea; Nitrogen; Soil; Methane; Nitrous Oxide; Agriculture; Fertilizers
PubMed: 37838042
DOI: 10.1016/j.scitotenv.2023.167629 -
Environmental Pollution (Barking, Essex... Mar 2024Isoprothiolane (IPT) and tricyclazole (TCZ) are widely used in rice farming and recently in combined rice-fish farming. However, co-cultured animals are affected by...
Risk assessment of pesticide compounds: IPT and TCZ cause hepatotoxicity, activate stress pathway and affect the composition of intestinal flora in red swamp crayfish (Procambarusclarkii).
Isoprothiolane (IPT) and tricyclazole (TCZ) are widely used in rice farming and recently in combined rice-fish farming. However, co-cultured animals are affected by these pesticides. To investigate the organismal effects and toxicity of pesticides, crayfish were exposed to 0, 1, 10, or 100 ppt TCZ or IPT for 7 days. Pesticide bioaccumulation, survival rate, metabolic parameters, structure of intestinal flora, and antioxidant-, apoptosis-, and HSP-related gene expression were determined. Pesticide exposure caused bioaccumulation of IPT or TCZ in the hepatopancreas and muscles of crayfish; however, IPT bioaccumulation was higher than that of TCZ. Both groups showed significant changes in hepatopancreatic serum biochemical parameters. Mitochondrial damage and chromosomal agglutination were observed in hepatopancreatic cells exposed to 100 ppt IPT or TCZ. IPT induced more significant changes in serum biochemical parameters than TCZ. The results of intestinal flora showed that Vibro, Flavobacterium, Anaerorhabdus and Shewanella may have potential for use as a bacterial marker of TCZ and IPT. Antioxidant-, apoptosis-, and HSP-related gene expression was disrupted by pesticide exposure, and was more seriously affected by IPT. The results suggest that IPT or TCZ induce hepatopancreatic cell toxicity; however, IPT or TCZ content in dietary crayfish exposed to 1 ppt was below the food safety residue standard. The data indicated that IPT exposure may be more toxic than TCZ exposure in hepatopancreas and intestines and toxicity of organism are alleviated by activating the pathway of stress-response, providing an understanding of pesticide compounds in rice-fish farming and food safety.
Topics: Animals; Antioxidants; Pesticides; Gastrointestinal Microbiome; Astacoidea; Risk Assessment; Chemical and Drug Induced Liver Injury; Thiazoles; Thiophenes
PubMed: 38185353
DOI: 10.1016/j.envpol.2024.123315 -
Comparative Biochemistry and... Feb 2024The experiment was conducted to investigate the effects of Bisphenol S (BPS) on growth, physiological and biochemical indices, and the expression of ecdysteroid receptor...
The effects of bisphenol S exposure on the growth, physiological and biochemical indices, and ecdysteroid receptor gene expression in red swamp crayfish, Procambarus clarkii.
The experiment was conducted to investigate the effects of Bisphenol S (BPS) on growth, physiological and biochemical indices, and the expression of ecdysteroid receptor (ECR) of the red swamp crayfish (Procambarus clarkii). The gene encoding ECR was isolated from red swamp crayfish by homologous cloning and rapid amplification of cDNA ends (RACE). The ECR transcripts were 1757 bp long and encoded proteins of 576 amino acids. The quantitative real-time PCR (qRT-PCR) analysis showed that the ECR gene was expressed in various tissues under normal conditions, and the highest level was observed in the ovary and the lowest level was observed in the muscle (P < 0.05). Then, the experiment was designed with four different BPS concentrations (0, 1, 10, and 100 μg/L), BPS exposure for 14 days, three parallel groups, and a total of 240 red swamp crayfish. At 100 μg/L BPS, the survival rate, weight gain rate, and relative length rate were decreased significantly (P < 0.05). Malonaldehyde (MDA) content reached the highest level at 100 μg/L BPS. When BPS concentration was higher than 10 μg/L, the activities of superoxide dismutase (SOD) and catalase (CAT) were significantly lower than those of the control group (P < 0.05). The expression levels of the ECR gene in ovary, intestinal, gill, and hepatopancreas tissues were significantly increased after BPS exposure (P < 0.05). The ECR gene expression in ovaries and Y-organs was significantly higher than other groups in 10 μg/L BPS (P < 0.05). The expressions of the tumor necrosis factor -α (TNF-α) and interleukin-6 (IL-6) genes in the hepatopancreas gradually increased, and the highest expression was observed exposed in 100 μg/L BPS (P < 0.05). This research will provide novel insights into the health risk assessment of BPS in aquatic organisms.
Topics: Animals; Female; Astacoidea; Receptors, Steroid; Gene Expression
PubMed: 38061619
DOI: 10.1016/j.cbpc.2023.109811 -
Journal of Hazardous Materials Sep 2023Pesticides are important for agricultural development; however, animals involved in rice-fish farming absorb the pesticides used during the farming process. Thiamethoxam...
Pesticides are important for agricultural development; however, animals involved in rice-fish farming absorb the pesticides used during the farming process. Thiamethoxam (TMX) is extensively used in agriculture and is gradually occupying the market for traditional pesticides. Therefore, this study aimed to investigate whether selenomethionine (SeMet) could affect the survival rate, bioaccumulation of TMX, serum biochemical parameters, lipid peroxidation, antioxidants in the hepatopancreas, and expression of stress genes after exposure of red swamp crayfish to 10 ppt TMX for 7 days. The results showed that the survival rate significantly increased and the bioaccumulation of TMX significantly decreased with SeMet administration (P < 0.05). Furthermore, severe histological damage to the hepatopancreas of red crayfish was observed after exposure to TMX; however, this damage was alleviated after SeMet administration. SeMet also significantly reduced the TMX-induced changes in serum biochemical parameters, malondialdehyde content, and antioxidant enzyme activity in crayfish hepatopancreas (P < 0.05). Notably, analysis of the expression of 10 stress response genes showed that 0.5 mg/kg SeMet might decrease cell damage in the hepatopancreas. Consequently, our findings suggest that higher levels of TMX in crayfish may cause hepatopancreatic cell toxicity, which can be harmful to human health; however, SeMet could mitigate these effects, providing an understanding of pesticide compounds and food safety.
Topics: Humans; Animals; Antioxidants; Selenium; Astacoidea; Thiamethoxam; Oxidative Stress; Bioaccumulation; Selenomethionine; Pesticides
PubMed: 37364439
DOI: 10.1016/j.jhazmat.2023.131896 -
Journal of Invertebrate Pathology Nov 2023The crayfish plague, caused by the pathogen Aphanomyces astaci, is a pandemic disease endemic to North America that has been devastating susceptible crayfish populations...
The crayfish plague, caused by the pathogen Aphanomyces astaci, is a pandemic disease endemic to North America that has been devastating susceptible crayfish populations in Europe since the 19th century. In Spain, this disease has decimated populations of the native crayfish species Austropotamobius pallipes due to introductions of North American crayfish, which act as vectors of the pathogen. To combat against these losses, several regional governments have established ex-situ breeding programs to restock wild populations of the species. In this study, we report on an outbreak of A. astaci that occurred in one of the most important A. pallipes aquaculture centers in Spain. Using a variety of detection methods, we analyzed affected crayfish and environmental samples from the facilities over a period of six months and determined that the outbreak was caused by two haplotypes of A. astaci, d1 and d2, which are both associated with the North American crayfish species Procambarus clarkii. To our knowledge, this is the first report of a two-haplotype coinfection of A. astaci outside the native range of this pathogen.
Topics: Animals; Haplotypes; Astacoidea; Aphanomyces; Europe; Disease Outbreaks
PubMed: 37659741
DOI: 10.1016/j.jip.2023.107989