RESUMEN
Aeromonas veronii is an important pathogen found in various aquatic environments and products, posing a threat to public health. The Hanks-like serine/threonine protein kinase is closely linked to the pathogenesis of pathogenic bacteria, but the exact role of YihE in A. veronii remains still unknown. To study the specific function of the YihE kinase, we constructed a knockout mutant of the yihE gene in A. veronii. The deletion of the yihE gene resulted in changes to the metabolism of L-arginine-AMC and acetic acid, as well as enhanced resistance to ampicillin and kanamycin in A. veronii. Additionally, the ΔyihE strain demonstrated a 1.4-fold increase in biofilm formation ability and a 1.8-fold decrease in adhesion and invasion to EPCs when compared to the wild-type strain. A significant decrease in cytotoxicity was observed at 2 and 3 h post-infection with EPCs compared to the wild-type strain. Additionally, the deletion of the yihE gene was associated with a significant decrease in motility of the strain. Furthermore, the deletion of the yihE gene resulted in a 1.44-fold increase in the LD50 of A. veronii in zebrafish. These findings offer valuable insights into the pathogenic mechanisms of A. veronii.
Asunto(s)
Aeromonas veronii , Adhesión Bacteriana , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Pez Cebra , Aeromonas veronii/patogenicidad , Aeromonas veronii/genética , Aeromonas veronii/fisiología , Virulencia , Animales , Enfermedades de los Peces/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Bacterias Gramnegativas/microbiología , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Biopelículas/crecimiento & desarrolloRESUMEN
The efficacy of phoxim in treating bacterial sepsis in silver carp is significant, yet its underlying mechanism remains elusive. This study aimed to establish a model of Aeromonas veronii infection in silver carp and subsequently treat the infected fish with 10 µg/L phoxim. Kidney and intestine samples from silver carp were collected for transcriptome analysis and assessment of intestinal microbial composition, with the aim of elucidating the mechanism underlying the efficacy of phoxim in treating bacterial sepsis in silver carp. The results of transcriptome and intestinal microbial composition analysis of silver carp kidney indicated that A. veronii infection could up-regulate the expression of il1ß, il6, nos2, ctsl, casp3 et al., which means, signifying that the kidney of silver carp would undergo inflammation, induce apoptosis, and alter the composition of intestinal microorganisms. Phoxim immersion might enhance the energy metabolism of silver carp and change its intestinal microbial composition, potentially elevating the antibacterial infection resistance of silver carp. These findings may contribute to an understanding of how phoxim can effectively treat bacterial sepsis in silver carp.
Asunto(s)
Carpas , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Compuestos Organotiofosforados , Animales , Carpas/inmunología , Enfermedades de los Peces/inmunología , Compuestos Organotiofosforados/farmacología , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/tratamiento farmacológico , Aeromonas veronii/fisiología , Microbioma Gastrointestinal/efectos de los fármacosRESUMEN
There is a long-standing debate on the attributes of temperature for fish health. We recently showed that thermoregulatory programs exerted through natural behavioural fever drive molecular and cellular responses that contribute to pathogen clearance, inflammation control, and tissue repair. These offered a mechanistic basis for the survival advantage conferred through fever. Herein, we show the attributes of mechanical replication of this fever response. Central to our approach was consideration of both, the maximal temperatures naturally selected by fish after infection, as well as the dynamics of thermal changes induced through this response. Coarse replication of the febrile thermal program as well as shorter truncated thermal schedules offered immune-regulatory capacity. Most notably, these promoted induction of acute inflammation and significant enhancements to pathogen clearance. However, the coarse protocols tested only partially recapitulated enhancements to induction and control of tissue repair. Our findings highlight a promising new alternative to combat infections in fish using a natural, drug-free, sustainable approach.
Asunto(s)
Aeromonas , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Animales , Aeromonas veronii/fisiología , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Bacterias Gramnegativas/prevención & control , Peces , Inflamación , Enfermedades de los Peces/prevención & control , Aeromonas/fisiologíaRESUMEN
In this study, the effects of Coriandrum sativum to control Aeromonas veronii infection in Oreochromis niloticus were determined. Coriandrum sativum extract (CE) was tested in vitro against A. veronii by the disc diffusion assay. In in vivo, 150 O. niloticus (from El-Abbassa, Sharkia, Egypt, weighing 34.95 ± 1.98 g) was distributed in five groups (with three replications) in glass aquariums (80 × 40 × 30 cm). The first group (control) was intraperitoneally injected with 0.2 ml of sterilized tryptic soya broth. Groups 2-5 were intraperitoneally challenged with 0.2 ml of A. veronii (4.3 × 106). The five groups were administered a basal diet until clinical signs appeared, and then therapeutic feeding (15 days) was followed: the first (CONT) and second (AV) groups were administered a normal basal diet. The third (AV+CP) and fourth (AV+CE) groups were administered diets supplemented with C. sativum powder and extract, respectively, each at 30 mg/kg. The fifth group (AV+OT) was administered a diet supplemented with oxytetracycline at 500 mg/kg diet. The results of the in vitro experiment revealed that CE has a zone of inhibition of 43 mm against A. veronii. The in vivo results showed that fish administered a therapeutic diet supplemented with CE showed a significant improvement in hematological, biochemical, and immunological parameters, as well as antioxidant capacity (P < 0.05) and the pathological findings of the liver and kidney tissues. The current findings supported that the administration of a CE-enriched diet (30 mg/kg) is an eco-friendly strategy for controlling A. veronii in O. niloticus.
Asunto(s)
Cíclidos , Coriandrum , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Animales , Antioxidantes/farmacología , Aeromonas veronii/fisiología , Dieta/veterinaria , Suplementos Dietéticos , Resistencia a la Enfermedad , Riñón/fisiología , Enfermedades de los Peces/prevención & control , Alimentación Animal/análisis , Infecciones por Bacterias Gramnegativas/prevención & control , Infecciones por Bacterias Gramnegativas/veterinariaRESUMEN
Aeromonas veronii is a pathogenic bacterium associated with various diseases in aquaculture. However, few studies address the antibacterial activity using nanoparticles (NPs). Hence, the current study is innovative to evaluate the antibacterial efficacy of silica nanoparticles (SiNPs) against A. veronii infection in-vitro with a trial for treatment in-vivo. Primarily, we assessed the in-vitro antibacterial activity against A. veronii. Further, we investigated the hematological profile, immune-antioxidant response, and gene expression of African catfish (Clarias gariepinus) in response to SiNPs exposure and the A. veronii challenge. Fish (N = 120; weight: 90 ± 6.19 g) were distributed into four groups (30 fish/group) for a ten-days-treatment trial. The first (control) and second (SiNPs) groups were treated with 0 mg/L and 20 mg/L SiNPs in water, respectively. The third (A. veronii) and fourth (SiNPs + A. veronii) groups were treated with 0 mg/L and 20 mg/L SiNPs in water, respectively, and infected with A. veronii (1.5 × 107 CFU/mL). Results demonstrated that SiNPs displayed an in-vitro antibacterial activity against A. veronii with a 21 mm inhibitory zone. A. veronii infection caused a high mortality rate (56.67%) and substantial reductions in hematological indices and immune indicators [nitric oxide (NO) and immunoglobulin M (IgM)]. Additionally, marked decline in the level of antioxidants [superoxide dismutase (SOD), catalase (CAT), and reduced glutathione content (GSH)] as well as down-regulation in the immune-related genes [interleukins (IL-1ß and IL-8) and tumor necrosis factor-alpha (TNF-α)] and antioxidant-related genes [SOD1, glutathione peroxidase (GPx), and glutathione-S-transferase (GST)] were the consequences of A. veronii infection. Surprisingly, treatment of A. veronii-infected fish with SiNPs lessened the mortality rate, enhanced the blood picture, modulated the immune-antioxidant parameters, and resulted in gene up-regulation. Overall, this study encompasses the significant role of SiNPs, a new versatile tool for combating hematological, immuno-antioxidant alterations, and gene down-regulation induced by A. veronii infection and sustainable aquaculture production.
Asunto(s)
Bagres , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Nanopartículas , Animales , Antioxidantes/metabolismo , Aeromonas veronii/fisiología , Regulación hacia Abajo , Expresión Génica , Bagres/genética , Bagres/metabolismoRESUMEN
Interferon regulatory factors (IRFs) play an important role in innate and adaptive immune system. However, in teleosts, the data on IRFs is still scarce. Here, for the first time, we identified 11 members of IRFs from the zig-zag eel Mastacembelus armatus (MarIRF1-10). The deduced protein sequences are highly conserved among different fish species especially in DBD and IAD domain. Phylogenetic analysis indicated that MarIRFs preferentially grouped with fish species in Synbranchiformes or Perciformes. Expression analysis showed that MarIRFs were expressed in all nine tissues including spleen, gill, muscle and intestine. After infected by Aeromonas veronii, expression of MarIRF2, MaIRF4b and MaIRF5 were significantly upregulated in spleen, MarIRF1, MarIRF2 were significantly upregulated in kidney, but in liver, nearly all MarIRFs were downregulated. Taken together, this study first reported molecular characterization and expression patterns of 11 IRFs in the zig-zag eel. All these results will contribute a lot to better understanding the antibacterial mechanism of IRFs in teleosts.
Asunto(s)
Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Animales , Factores Reguladores del Interferón/metabolismo , Aeromonas veronii/fisiología , Filogenia , Peces/genética , Anguilas/genética , Proteínas de Peces/metabolismo , Factor 1 Regulador del Interferón/metabolismoRESUMEN
The potential of rice protein concentrate (RPC) to substitute fishmeal (FM) protein in the diet of Oreochromis niloticus was assessed in a five-month-long feeding trial. Fishmeal protein was replaced by RPC at rates of 0% (control), 25%, 50%, and 75% (RPC0, RPC25, RPC50, and RPC75, respectively). RPC25 had no significant effect on antioxidant capacity (total antioxidant capacity; superoxide dismutase, catalase, and glutathione peroxidase activities) and immune indices (lysozyme, nitric oxide, antiprotease, and bactericidal activities) after one, two, and five months of feeding, while the values for these parameters were significantly lower in the RPC75 group compared to those in the RPC0 group. The RPC25 group showed higher mRNA levels of the intestinal cytokines IL-1ß, IL-10ß, TGF-ß, and TNF-α than the control group. In fish affected by Aeromonas veronii, the highest significant cumulative mortality was recorded in the RPC75 group, followed by the RPC50, RPC25, and control groups. Gut microbiome analyses showed a reduction in microbial diversity in response to the addition of RPC, regardless of the RPC content, and the composition of the community of the RPC samples differed from that of the control. RPC-enriched diets resulted in higher relative abundances of Bacteroidetes and Fusobacteria in the gut compared to that in the gut of the control fish. In summary, RPC can be used to replace up to 25% of the FM protein in the diet of O. niloticus, while improving the antioxidant capacity, immunocompetence, and disease resistance of the fish.
Asunto(s)
Cíclidos , Enfermedades de los Peces , Microbioma Gastrointestinal , Infecciones por Bacterias Gramnegativas , Oryza , Aeromonas veronii/fisiología , Alimentación Animal/análisis , Animales , Antioxidantes/metabolismo , Citocinas , Dieta/veterinaria , Resistencia a la EnfermedadRESUMEN
In November 2019, an acute disease outbreak in Australian redclaw crayfish (Cherax quadricarinatus) occurred in a farm in Hubei, China, with a cumulative mortality rate of over 80%. One of the characteristic symptoms of the disease was blisters on the tail. This symptom is also common in diseased Procambarus clarkii every year in this country, but the causative agent has not been determined. This study analyzed the etiological characteristics of this disease. Bacterial isolation and identification combined with high-throughput sequencing analysis were conducted to obtain the microbiota characteristics in the hemolymph, hepatopancreas, and intestines. Results showed that this outbreak was caused by infection from Aeromonas hydrophila and Aeromonas veronii. The underlying cause was stress imposed on crayfish during transferring from outdoor pond to indoor pond because of temperature drops. Aeromonas infection caused remarkable changes in the structure of the microbial composition in the hemolymph, hepatopancreas, and intestines of the crayfish. The abundance of Aeromonas in the hemolymph of the sick crayfish was as high as 99.33%. In particular, KEGG metabolic pathway analysis showed that some antibiotic synthesis, enterobactin biosynthesis, and myo-inositol degradation pathways were abundant in healthy crayfish hemolymphs, which may be the mechanism of maintaining crayfish health. Conversely, inhibition of these pathways led to the disorder of microbiota structure, finally leading to the occurrence of diseases. To the knowledge of the authors, this study was the first to use high-throughput amplicon sequencing targeting the 16S rRNA gene to find the causative bacteria in aquatic animals. This protocol can provide more comprehensive and reliable evidence for pathogen identification, even if the pathogenic bacteria are anaerobes or other hard-to-culture bacteria.
Asunto(s)
Aeromonas hydrophila/fisiología , Aeromonas veronii/fisiología , Astacoidea/microbiología , Animales , China , Hemolinfa/microbiología , Hepatopáncreas/microbiología , Intestinos/microbiología , Cola (estructura animal)/microbiología , Cola (estructura animal)/patologíaRESUMEN
Bacterial disease outbreaks in filter feeder bivalve Hyriopsis cumingii as water contamination become more frequent in the water ecosystem, especially in intensive aquaculture habitats. To characterize host-pathogen interactions between H. cumingii and bacterial infection, we investigated the effects of Stenotrophomonas maltophilia HOP3 and Aeromonas veronii GL1 on the antioxidant response, tissue invasion and transcriptome expression of H. cumingii by infectivity trials. We showed that bacterial infections resulted in tubular necrosis of the hepatopancreas and induced the acute immune response in H. cumingii. The transcriptomic study identified a total of 5957 differentially expressed genes (DEGs) after A. veronii challenge. These DEGs were implicated in 302 KEGG pathways, notably in Apoptosis, Phagosome and Lysosome. The results showed that the relative expressions of all six immune-related DEGs were effectively stimulated with A. veronii, accompanied by tissue differences. Overall, these findings will contribute to an analysis of the immune response of H. cumingii to bacterial infection at the transcriptomic level and its genomic resource for research.
Asunto(s)
Expresión Génica/inmunología , Infecciones por Bacterias Gramnegativas/inmunología , Transcriptoma/inmunología , Unionidae/inmunología , Aeromonas veronii/fisiología , Animales , Antioxidantes/metabolismo , Acuicultura , Infecciones por Bacterias Gramnegativas/genética , Infecciones por Bacterias Gramnegativas/patología , Hepatopáncreas/inmunología , Hepatopáncreas/patología , Interacciones Huésped-Patógeno/inmunología , Stenotrophomonas maltophilia/fisiología , Distribución Tisular , Unionidae/genética , Unionidae/microbiología , Factores de Virulencia/inmunologíaRESUMEN
Bacillus amyloliquefaciens X030 (BaX030) has broad-spectrum antibacterial activity against the fish pathogens Aeromonas hydrophila and Aeromonas veronii. To improve its antibacterial effect, BaX030 was subjected to compound mutagenesis of atmospheric and room temperature plasma (ARTP) and nitrosoguanidine (NTG). The results showed that, compared with the original strain, the production of macrolactin A and oxydifficidin in mutated strain N-11 increased to 39 % and 268 %, respectively. The re-sequencing analysis suggested that there were SNPs and InDels in the gene clusters focused on the sucrose utilization pathway, glycolysis pathway and fatty acid synthesis pathway. Scanning electron microscopy revealed that strain N-11 became thin and long. The qRT-PCR results indicated that the expression of immune factors in the liver or kidney tissue of grass carp increased after feeding with N-11. H&E staining and protection experiments also showed that the mortality and surface symptoms of grass carp infected by the two pathogens were significantly reduced. The study identified a probiotic strain with potential application value in aquaculture production and provided a new strategy for the discovery of new strains with higher antibacterial biological activity.
Asunto(s)
Aeromonas hydrophila/fisiología , Aeromonas veronii/fisiología , Bacillus amyloliquefaciens/genética , Carpas/microbiología , Interacciones Microbianas/genética , Mutación , Probióticos , Animales , Bacillus amyloliquefaciens/fisiología , Enfermedades de los Peces/microbiologíaRESUMEN
The heavy use of prophylactic antibiotics in aquaculture leads to elevated antibiotic residues, posing a huge hidden danger in aquaculture products and other natural aquatic environments. Therefore, this study aims to isolate probiotics that can replace antibiotics from the gut of grass carp for disease control. Bacillus velezensis B8 was isolated from the gut of grass carp and showed broad-spectrum antimicrobial activity against several fish pathogenic bacteria, including Aeromonas hydrophilis, Aeromonas veronii, Vibrio parahaemolyticus, Escherichia coli, Edwardsiella tarda and Vibrio mimicus. The safety evaluation showed that the strain B8 was non-toxic to grass carp, had no hemolytic activity, and was sensitive to most antibiotics. In vitro study indicated that strain B8 was viable at pH 2-7, had weak tolerance to 0.1% (w/v) bile salt, and could grow at 10°C-40 °C. The grass carps were fed with diets containing 0 (control), 107, and 109 cfu/g of strain B8 for 4 weeks. Various immune parameters were measured at 1, 2, 3, and 4 weeks of post-feeding. The results of non-specific immunoassay showed that diets supplemented with B8 significantly increased alkaline phosphatase (AKP) and superoxide dismutase (SOD) activity in serum samples (p < 0.05). The expression levels of immune-related genes in the kidney and spleen of grass carp were measured. Among them, the expression levels of IgM and TNF-α both in spleen and kidney were significantly increased after 3 and 4 weeks of post-feeding (p < 0.05). The expression of IgD and MHCI in kidney was significantly upregulated in high-dose groups after 2 and 3 weeks of feeding, respectively (p < 0.05). In addition, after 7 days of challenging with A. veronii, the high-dose group and low-dose group had 48% and 53% survival compared to 25% survival for the control group. These results suggest that B. velezensis B8 has the potential to be developed into a microecological preparation for the alternatives of antibiotics in aquaculture.
Asunto(s)
Bacillus/química , Carpas/inmunología , Resistencia a la Enfermedad , Enfermedades de los Peces/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Inmunidad Innata , Probióticos/metabolismo , Aeromonas veronii/fisiología , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Resistencia a la Enfermedad/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Infecciones por Bacterias Gramnegativas/inmunología , Inmunidad Innata/efectos de los fármacos , Probióticos/administración & dosificación , Distribución AleatoriaRESUMEN
The oriental river prawn Macrobrachium nipponense is a commercially important freshwater shrimp that is widely farmed in China. Aeromonas veronii is a conditional pathogen of farmed shrimp, which has caused huge economic losses to the industry. Therefore, there is urgency to study the host-pathogen interactions between M. nipponense and A. veronii to screen individuals with antimicrobial resistance. In this study, we examined the hepatopancreas of moribund M. nipponense infected with A. veronii and healthy individuals at both the histopathological and transcriptomic levels. We showed that A. veronii infection resulted in tubular necrosis of the M. nipponense hepatopancreas. Such changes likely affect assimilation, storage, and excretion by the hepatopancreas, which could ultimately affect the survival and growth of infected individuals. Among the 61,345 unigenes obtained through RNA sequencing and de novo transcriptome assembly, 232 were differentially expressed between the two groups. KEGG and GO analyses revealed that these differentially expressed genes were implicated in pathways, including PPAR, PI3K/AKT, and AMPK signaling. The results of this study will contribute to an analysis of the immune response of M. nipponense to A. veronii infection at the transcriptomic level. Furthermore, the RNA-seq data generated here provide an important genomic resource for research on M. nipponense in the absence of a reference genome.
Asunto(s)
Aeromonas veronii/fisiología , Hepatopáncreas/inmunología , Palaemonidae/microbiología , Alimentos Marinos/microbiología , Transcriptoma/inmunología , Animales , China , Hepatopáncreas/patología , Interacciones Huésped-Patógeno , Necrosis , Palaemonidae/inmunología , Transducción de SeñalRESUMEN
The current work was planned to assess the effects of dietary microalga, Nannochloropsis oculata (NP) on the growth indices, serum biochemistry, non-specific immunity, and resistance of Nile tilapia (Oreochromis niloticus) juveniles against challenge with pathogenic Aeromonas veronii. Fish (10.21 ± 0.28 g) were randomly divided into four treatments in triplicates and were fed on diets supplemented with different levels of NP (0, 5, 10, and 15%) for eight weeks. The results showed that the dietary 5% NP significantly improved the growth parameters (final body weight, weight gain, specific growth rate, and total feed intake) (P < 0.05). Meanwhile, the feed conversion ratio was significantly decreased in NP-supplemented groups with respect to the control group. The serum protein profile (total protein, albumin, globulin, and albumin/globulin ratio) was significantly decreased by 15% NP supplemented group. Meanwhile, liver enzymes (alanine transaminase, and aspartate transaminase) activities were not significantly affected by NP dietary supplementation (P < 0.05). Serum lysozyme activity, nitric oxide (NO), and nitroblue tetrazolium (NBT) levels were significantly enhanced only in 5% NP supplemented group (P < 0.05). Interestingly, the hepatopancreatic and intestinal tissues had apparently normal histomorphology of Nile tilapia fed at 5% NP-supplemented diets. Significant upregulation of cytokines [interleukin 1beta (IL-1ß), interleukin 8 (IL-8), interferon-gamma (IFN-γ), transforming growth factor-beta (TGF- ß), and tumor necrosis factor-alpha (TNF-α)], with significant downregulation of the antioxidant gene [superoxide dismutase (SOD)] in 10% and 15% NP supplemented groups. Attractively, the relative level of protection (RLP) against challenge with pathogenic A. veronii was significantly elevated in 10% and 15% NP supplemented groups. Conclusively, the obtained results reflect the beneficial roles of dietary NP to improve growth and boost the immune responses of Nile tilapia.
Asunto(s)
Cíclidos/inmunología , Resistencia a la Enfermedad/inmunología , Enfermedades de los Peces/inmunología , Estramenopilos/química , Aeromonas veronii/fisiología , Alimentación Animal/análisis , Animales , Cíclidos/sangre , Cíclidos/crecimiento & desarrollo , Dieta/veterinaria , Suplementos Dietéticos/análisis , Relación Dosis-Respuesta a Droga , Enfermedades de los Peces/microbiología , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Distribución AleatoriaRESUMEN
The present study investigated the effects of black mulberry (Morus nigra) syrup supplementation on growth performance, hematological, serum biochemical, innate immune parameters, immune and antioxidant related gene expression responses, and disease resistance of Nile tilapia, Oreochromis niloticus. Five isonitrogenous and isoenergetic diets were formulated to contain black mulberry syrup at levels of 0%, 0.75%, 1.5%, 2.0%, and 3.0%. Fish were fed experimental diets for 60 days. Dietary black mulberry syrup increased activities of serum lysozyme, myeloperoxidase, superoxide dismutase and catalase, and increased the expression levels of immune-related genes (interleukin 1, beta, tumor necrosis factor, immunoglobulin M, interferon gamma and heat shock protein 70) in the spleen and antioxidant-related genes (superoxide dismutase, catalase and glutathione peroxidase) in the liver of fish fed especially with 1.5%, 2.0%, and 3.0% black mulberry syrup supplemented diets. Furthermore, at 2.0% incorporation level, growth performance increased. The findings of the present study indicate that Nile tilapia fed with diet containing 2.0% black mulberry might be adequate to improve the growth performance, innate immune parameters, antioxidant related gene expression responses, and disease resistance against Aeromonas veronii.
Asunto(s)
Cíclidos/inmunología , Resistencia a la Enfermedad , Enfermedades de los Peces/inmunología , Proteínas de Peces/genética , Expresión Génica/inmunología , Morus/química , Extractos Vegetales/metabolismo , Aeromonas veronii/fisiología , Alimentación Animal/análisis , Animales , Cíclidos/genética , Cíclidos/crecimiento & desarrollo , Cíclidos/fisiología , Dieta/veterinaria , Suplementos Dietéticos/análisis , Resistencia a la Enfermedad/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Proteínas de Peces/metabolismo , Expresión Génica/efectos de los fármacos , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Extractos Vegetales/administración & dosificación , Distribución AleatoriaRESUMEN
Lactic acid bacteria (LAB) has been documented to promoting growth, enhancing immunity and disease resistance. In this study, we aimed to evaluate the single or conjoint effects of Lactococcus lactis L19 (Genbank: MT102745.1) and Enterococcus faecalis W24 (Genbank: MT102746.1) isolated from the intestine of Channa argus (C. argus) on growth performance, immune response and disease resistance of C. argus. A total of 720 apparently healthy C. argus (9.50 ± 0.03 g) were randomly divided into four equal groups. Fish were fed with a basal diet (CK) supplemented with L. lactis (L19), E. faecalis (W24), and L. lactis L19 + E. faecalis W24 (L + W) at 1.0 × 108 cfu/g basal diet for 56 days. After feeding, the final body weight (FBW), weight gain (WG), feed efficiency ratio (FER), specific growth rate (SGR) and protein efficiency ratio (PER) had significantly increased (p < 0.05), especially with L19. The results indicated that single or conjoint administration of LAB as potential probiotics can induce high levels of IgM, ACP, AKP, LZM, C3 and C4 activity in serum, which may effectively induce humoral immunity, and L19 induce even higher levels. Meanwhile, when compared to CK group, the results of qPCR showed that LAB administration significantly up-regulated (p < 0.05) the expression of IL-1ß, IL-6, IL-10, TNF-α, IFN-γ, HSP70, HSP90, TGF-ß in the spleen, head kidney, gill, liver and intestine of C. argus. After challenge with Aeromonas veronii, the survival rates in all LAB-fed groups were significantly higher (p < 0.05) than that of the CK group, and the L19 group showed the highest (63.3%) disease resistance. Our data indicated that L. lactis L19 and E. faecalis W24, as a feed additive at 1.0 × 108 cfu/g feed, could promote growth performance, enhance immune response and disease resistance of C. argus, with greatest effects in fish fed L. lactis L19 for 56 days. Hence, these LAB additives could be used as promising probiotics for C. argus. L19 was more effective than W24 or the mixture of the two for promoting growth performance, enhancing immune response and disease resistance of C. argus.
Asunto(s)
Resistencia a la Enfermedad/efectos de los fármacos , Enterococcus faecalis/química , Enfermedades de los Peces/inmunología , Peces/inmunología , Inmunidad Humoral/efectos de los fármacos , Lactococcus lactis/química , Probióticos/metabolismo , Aeromonas veronii/fisiología , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Peces/crecimiento & desarrollo , Microbioma Gastrointestinal/fisiología , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Intestinos/efectos de los fármacos , Intestinos/microbiología , Probióticos/administración & dosificación , Distribución AleatoriaRESUMEN
Aeromonas veronii is a widely distributed novel pathogen that can affect humans and animals, it can cause sepsis in fish with high mortality and serious economic losses to aquaculture. In the study, the gut microbiome of the infected and uninfected grass carp with Aeromonas veronii were analyzed probiotics and pathogenic bacteria by the Miseq high-throughput sequencing, the results showed that the infected fish were significantly higher in Proteobacteria, Firmicutes, Fusobacteria, and the immune factors in liver and kidney were up-regulated by qRT-PCR. In order to effectively inhibit the pathogen, we screened an actinomycete strain and had good antibacterial effect on Aeromonas veronii. The new antagonistic bacteria was named as Streptomyces flavotricini X101, the whole genome sequencing revealed that the metabolic process was most active. After grass carp was inoculated with the minimum inhibitory concentration of 900 µg/mL of the strain's fermentation supernatant, then Aeromonas veronii was injected, we found that the pathological symptoms such as body surface, anus and abdominal congestion were alleviated by H&E staining. Cellular experiments showed that it wasn't toxic to liver cells of grass carp. Overall, this is the first study of changes in intestinal flora, phenotype, and immune factors in grass crap infected with Aeromonas veronii, it had important theoretical significance and application value for immunization and prevention.
Asunto(s)
Aeromonas veronii/fisiología , Carpas/microbiología , Enfermedades de los Peces/microbiología , Microbioma Gastrointestinal , Infecciones por Bacterias Gramnegativas/veterinaria , Streptomyces/fisiología , Animales , Carpas/inmunología , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/patología , Microbioma Gastrointestinal/genética , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/microbiología , Infecciones por Bacterias Gramnegativas/patología , Secuenciación de Nucleótidos de Alto Rendimiento , Inmunoglobulina M/metabolismo , Interleucinas/metabolismo , ARN Ribosómico 16S/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Streptomyces/genéticaRESUMEN
Aeromonas veronii is a virulent fish pathogen that causes extensive economic losses in the aquaculture industry worldwide. In this study, a virulent strain of A. veronii TH0426 was used to establish an in vitro biofilm model. The results show that the biofilm-forming abilities of A. veronii TH0426 were similar in different media, peaking under conditions of 20 °C and pH 6. Further, isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics methods were used to compare the differential expression of A. veronii between the biofilm and planktonic cells. The results show alterations in 277 proteins, with 130 being upregulated and 147 downregulated. Pathway analysis and GO (Gene Ontology) annotations indicated that these proteins are mainly involved in metabolic pathways and the biosynthesis of secondary metabolites and antibiotics. These proteins are the main factors affecting the adaptability of A. veronii to its external environment. MRM (multiple reaction 27 monitoring) and qPCR (qPCR) were used to verify the differential proteins of the selected A. veronii. This is the first report on the biofilm and planktonic cells of A. veronii, thus contributing to studying the infection and pathogenesis of A. veronii.
Asunto(s)
Aeromonas veronii/fisiología , Proteínas Bacterianas/biosíntesis , Biopelículas/crecimiento & desarrollo , Regulación Bacteriana de la Expresión Génica/fisiología , ProteómicaRESUMEN
Suppressors of cytokine signaling (SOCS) family members have negative effects on cytokine signaling pathways involved in immunity, growth and development. Owing to their typical feature, they have been extensively studied in mammalians, but they have not offered systematic studies among teleosts. In the present study, nine SOCS family genes were identified in the swamp eel genome and analyzed regulation mechanisms of SOCS family members in swamp eels. The open reading frames of MaSOCS1a, MaSOCS1b, MaSOCS2, MaSOCS3a, MaSOCS3b, MaSOCS4, MaSOCS5, MaSOCS6 and MaSOCS7 were 663 bp, 603 bp, 717 bp, 618 bp, 645 bp, 1188 bp, 1488 bp, 1611 bp and 1998 bp and encoded 220, 238, 200, 205, 214, 395, 496, 536 and 655 amino acids, respectively. All SOCS proteins have no signal peptides. Multiple alignment revealed that MaSOCS family members possessed a typical conserved SOCS box and SH2 region. Phylogenetic analyses showed that all SOCS proteins were divided into two main clusters. Taken together with the similarity and identity of SOCS protein amino acids, these results indicated that MaSOCS family members shared conserved with other homologous genes, in which MaSOCS7 was more conserved. Further syntenic analysis confirmed the phylogenetic analysis results and annotation of SOCS protein, suggesting that MaSOCS5 shared a common ancestor gene with that of fish and humans. MaSOCS family members were constitutively expressed in a wide range of tissues with different levels. In particular, spleen and head kidneys play an important role in immune-related pathways. After Aeromonas veronii and polyinosinic-polycytidylic acid (poly I:C) challenge in the spleen and head kidney, MaSOCS family members exhibit different expression profiles. These expression patterns indicated that MaSOCS family members could make acute responses after pathogen invasion. Taken together, these results indicate that MaSOCS family members participate in the immune response against pathogens and offer a solid foundation for future studies of SOCS function.
Asunto(s)
Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Smegmamorpha/genética , Smegmamorpha/inmunología , Proteínas Supresoras de la Señalización de Citocinas/genética , Proteínas Supresoras de la Señalización de Citocinas/inmunología , Aeromonas veronii/fisiología , Animales , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Filogenia , Poli I-C/farmacologíaRESUMEN
The effects of dietary administration of Astragalus membranaceus nanoparticles (ANP) on immune and anti-oxidative responses, growth performance and disease resistance of Oreochromis niloticus were evaluated in the present study. Fish were divided into three groups and received the ANP at rates of 0 (control), 1, and 2%/kg diet for four weeks. After the four-week feeding trial, three fish from each replicate were sampled for immune and anti-oxidative responses evaluation, ten fish from each group were challenged with A. veronii, and nine fish from each group were subjected to cold and hypoxia challenges. It was obvious from the results that ANP significantly enhanced lysozyme activity and nitrous oxide (NO) activities, as well as improved superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities. Also, aspartate aminotransferase, alanine transaminase, glucose, and cortisol measurements showed significantly lower levels in incorporated groups compared to the control. Growth performance; and amylase and lipase digestive enzymes activities also showed markedly improved results. Expression of heat shock protein 70 (HSP70) and interleukin 1, beta (IL-1ß) genes were significantly upregulated throughout the entire experimental period. When challenged with A. veronii, the mortality of treated groups was significantly (P < 0.05) lower than the control. Current results proofs that dietary ANP had a synergistic effect on immune and anti-oxidative responses, growth performance and disease resistance of Oreochromis niloticus.
Asunto(s)
Antioxidantes/metabolismo , Astragalus propinquus/química , Cíclidos/inmunología , Enfermedades de los Peces/inmunología , Inmunidad Innata/efectos de los fármacos , Nanopartículas , Aeromonas veronii/fisiología , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Suplementos Dietéticos/análisis , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Nanopartículas/administración & dosificaciónRESUMEN
Interleukin-17 (IL-17) is an important cytokine that plays a critical role in the inflammatory response and host defense against extracellular pathogens. In the present study, six novel IL-17 family genes (MaIL-17) were identified by analyzing Asian swamp eel (Monopterus albus) genome. Sequence analysis revealed that the MaIL-17 family genes shared similar features, comprising a signal peptide, an IL-17 superfamily region, and four conserved cysteines. Phylogenetic analysis showed that the MaIL-17 genes were clustered together with their corresponding IL-17 genes from other species. The similarity and identity of all IL-17 family genes indicated that the MaIL-17 genes are conserved among teleosts, while Ma-IL-17D is more conserved than the other Ma-IL-17s. Except for MaIL-17A/F3 and MaIL-17D, all MaIL-17s shared the same genomic structure as the genes from other fish, namely three exons and two introns. The MaIL-17s showed conserved synteny among fish, and we found that the MaIL-17D locus has a more conserved syntenic relationship with the loci from other fish and humans. These results demonstrated that MaIL-17D and human IL-17D might have evolved from a common ancestral gene and subsequently diverged. The analysis of swamp eel reference genes revealed that EEF1A1 (encoding eukaryotic translation elongation factor 1 alpha 1) was an ideal reference gene for accurate real-time qRT-PCR normalization in the swamp eel. The MaIL-17 genes are widely distributed throughout tissues, suggesting that MaIL-17s carry out their biological functions in immune and non-immune tissues compartments. The transcript of Ma-IL17s exhibited different fold changes in head kidney cells in response to Aeromonas veronii phorbol 12-myristate 13-acetate (PMA) and polyinosinic:polycytidylic acid (poly I:C) challenge, showing that MaIL-17A/F1 has stronger antiviral activities compared with other MaIL-17 family genes, and that MaIL-17A/F3 and MaIL-17A/F2 possess stronger effects against extracellular pathogens compared with the others; however, MaIL-17C2 and MaIL-17D may play vital roles during pathogen infection. The differential immune responses of these genes to Aeromonas veronii, PMA and poly I:C implied distinct mechanisms of host defense against extracellular pathogens.