RESUMEN
Mammalian interleukin-22 (IL-22) attenuates organismal injury by inhibiting reactive oxygen species (ROS) and impeding the NLRP3 inflammasome activation. However, the role of fish IL-22 in this process remains unclear. We characterized MaIL-22, an IL-22 homolog in blunt snout bream (Megalobrama amblycephala). Despite its low sequence identity, it shares conserved structures and close evolutionary relationships with other teleost IL-22s. Furthermore, Aeromonas hydrophila (A. hydrophila) infection leads to tissue injury in M. amblycephala immune organs and concomitantly altered Mail-22 mRNA expression, suggesting that MaIL-22 was involved in the antimicrobial immune response. To explore MaIL-22's biological functions, we produced recombinant MaIL-22 (rMaIL-22) protein and demonstrated it significantly enhanced the survival of M. amblycephala post-A. hydrophila infection. To unravel its protective mechanisms, we explored the ROS/NLRP3 inflammasome axis and its downstream signaling responses. The results showed that rMaIL-22 treatment significantly elevated antioxidant enzyme (T-SOD, CAT and GSH-PX) activities to inhibit MDA activity and scavenge ROS in visceral tissues. Meanwhile, rMaIL-22 impeded the activation of NLRP3 inflammasome by suppressing NLRP3 protein and mRNA expression. This indicated that rMaIL-22 contributed to inhibit A. hydrophila-induced activation of the ROS/NLRP3 inflammasome axis. Consistent with these findings, rMaIL-22 treatment attenuated the expression of proinflammatory cytokines (il-1ß, tnf-α and il-6) and proapoptotic genes (caspase-3 and caspase-8) while promoting antiapoptotic genes (bcl-2b and mcl-1a) expression, ultimately mitigating tissue injury in visceral tissues. In conclusion, our research underscores MaIL-22's key role in microbial immune regulation, offering insights for developing IL-22-targeted therapies and breeding programs.
Asunto(s)
Aeromonas hydrophila , Apoptosis , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Inflamasomas , Inflamación , Interleucina-22 , Interleucinas , Proteína con Dominio Pirina 3 de la Familia NLR , Especies Reactivas de Oxígeno , Animales , Aeromonas hydrophila/inmunología , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Inflamasomas/metabolismo , Inflamasomas/inmunología , Apoptosis/efectos de los fármacos , Infecciones por Bacterias Gramnegativas/inmunología , Especies Reactivas de Oxígeno/metabolismo , Interleucinas/metabolismo , Interleucinas/inmunología , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Inflamación/inmunología , Proteínas de Peces/metabolismo , Proteínas de Peces/inmunología , Proteínas de Peces/genética , Cyprinidae/inmunología , Cyprinidae/metabolismo , Transducción de SeñalRESUMEN
Polymeric immunoglobulin receptor (pIgR) is an important immune factor in the mucosal immune system of fish, which plays a key role in mediating the secretion and transport of immunoglobulin into mucus. In this study, the full-length cDNA sequence of Megalobrama amblycephala pIgR gene was firstly cloned and the immune response to Aeromonas hydrophila was detected. After being challenged by Aeromonas hydrophila at 3 d, significantly pathological features were observed in intestine, head kidney, spleen, liver and gill of Megalobrama amblycephala. The content of lysozyme (Lys) and the activities of acid phosphatase (ACP) and alkaline phosphatase (AKP) increased significantly at 1 d and reached the peak at 3 d, and the activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-PX) and catalase (CAT) in serum reached the peak at 5 d and 7 d after infection, respectively. The expression level of IL-1ß gene reached the peak at 3 d in intestine, 5 d in gill and spleen, 7 d in head kidney and liver of Megalobrama amblycephala after infected by Aeromonas hydrophila, respectively. The TNF-α gene expression reached the peak at 3 d in intestine and gill, 5 d in head kidney and spleen, 7 d in liver after infection, respectively. The experimental results showed that the infection of Aeromonas hydrophila caused the pathological changes of immune-related tissues and triggered the inflammation responses. The full-length cDNA sequence of Megalobrama amblycephala pIgR was 1828 bp, and its open reading frame (ORF) was 1023 bp, encoding 340 amino acids. The pIgR of Megalobrama amblycephala has a signal peptide sequence, followed by extracellular region, transmembrane region and intracellular region. The extracellular region includes two Ig-like domains (ILDs), and its tertiary structure is twisted "L". The phylogenetic tree was constructed using the adjacency method, and the pIgR genes of Megalobrama amblycephala and cyprinidae fish were clustered into a single branch. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of pIgR gene in different tissues of Megalobrama amblycephala. The expression level of pIgR gene was the highest in liver, followed by intestine, head kidney, skin, middle kidney and spleen, lower in heart, gill and brain, and the lowest in muscle. After being infected by Aeromonas hydrophila, the expression level of Megalobrama amblycephala pIgR gene in intestine, head kidney, spleen, liver and gill showed a trend of increasing first and then decreasing within 28 d. The pIgR gene expression reached the peak in mucosal immune-related tissues (gill and intestine) was earlier than that in systemic immune-related tissues (head kidney and spleen), and the relative expression level of pIgR gene at peak in intestine (12.3 fold) was higher than that in head kidney (3.73 fold) and spleen (7.84 fold). These results suggested that Megalobrama amblycephala pIgR might play an important role in the mucosal immune system to against Aeromonas hydrophila infection.
Asunto(s)
Aeromonas hydrophila , Secuencia de Aminoácidos , Cyprinidae , Enfermedades de los Peces , Proteínas de Peces , Infecciones por Bacterias Gramnegativas , Inmunidad Innata , Filogenia , Receptores de Inmunoglobulina Polimérica , Animales , Aeromonas hydrophila/fisiología , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Cyprinidae/inmunología , Cyprinidae/genética , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Proteínas de Peces/química , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Receptores de Inmunoglobulina Polimérica/química , Inmunidad Innata/genética , Regulación de la Expresión Génica/inmunología , Alineación de Secuencia/veterinaria , Perfilación de la Expresión Génica/veterinaria , Secuencia de BasesRESUMEN
Introduction: STAT1a is an essential signal transduction protein involved in the interferon pathway, playing a vital role in IFN-alpha/beta and gamma signaling. Limited information is available about the STAT protein in fish, particularly in Indian major carps (IMC). This study aimed to identify and characterize the STAT1a protein in Labeo rohita (LrSTAT1a). Methods: The full-length CDS of LrSTAT1a transcript was identified and sequenced. Phylogenetic analyses were performed based on the nucleotide sequences. The in-vivo immune stimulant poly I: C was used to treat various tissues, and the expression of LrSTAT1a was determined using quantitative real-time polymerase chain reaction (qRT-PCR). A 3D model of the STAT1a protein was generated using close structure homologs available in the database and checked using molecular dynamics (MD) simulations. Results: The full-length CDS of Labeo rohita STAT1a (LrSTAT1a) transcript consisted of 3238 bp that encoded a polypeptide of 721 amino acids sequence was identified. Phylogenetic analyses were performed based on the nucleotide sequences. Based on our findings, other vertebrates share a high degree of conservation with STAT1a. Additionally, we report that the in vivo immune stimulant poly I: C treatment of various tissues resulted in the expression of LrSTAT1a as determined by quantitative real-time polymerase chain reaction (qRT-PCR). In the current investigation, treatment with poly I: C dramatically increased the expression of LrSTAT1a in nearly every organ and tissue, with the brain, muscle, kidney, and intestine showing the highest levels of expression compared to the control. We made a 3D model of the STAT1a protein by using close structure homologs that were already available in the database. The model was then checked using molecular dynamics (MD) simulations. Consistent with previous research, the MD study highlighted the significance of the STAT1a protein, which is responsible for Src homology 2 (SH2) recognition. An important H-bonding that successfully retains SH2 inside the STAT1a binding cavity was determined to be formed by the conserved residues SER107, GLN530, SER583, LYS584, MET103, and ALA106. Discussion: This study provides molecular insights into the STAT1a protein in Rohu (Labeo rohita) and highlights the potential role of STAT1a in the innate immune response in fish. The high degree of conservation of STAT1a among other vertebrates suggests its crucial role in the immune response. The in-vivo immune stimulation results indicate that STAT1a is involved in the immune response in various tissues, with the brain, muscle, kidney, and intestine being the most responsive. The 3D model and MD study provide further evidence of the significance of STAT1a in the immune response, specifically in SH2 recognition. Further research is necessary to understand the specific mechanisms involved in the IFN pathway and the role of STAT1a in the immune response of IMC.
Asunto(s)
Proteínas de Peces , Filogenia , Poli I-C , Factor de Transcripción STAT1 , Animales , Poli I-C/inmunología , Factor de Transcripción STAT1/metabolismo , Factor de Transcripción STAT1/genética , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Proteínas de Peces/metabolismo , Dominios Homologos src , Unión Proteica , Secuencia de Aminoácidos , Simulación de Dinámica Molecular , Carpas/inmunología , Carpas/genética , Carpas/metabolismo , Perfilación de la Expresión Génica , Cyprinidae/inmunología , Cyprinidae/genética , Cyprinidae/metabolismoRESUMEN
Bacterial extracellular vesicles (BEVs) are natural nanocarriers that have shown great potential for biomedical applications such as biomarkers, cancer therapy, immunomodulators, vaccines, wound healing, tissue engineering, and drug carriers. In the present study, BEVs were isolated from the gram-negative bacterium, Aeromonas hydrophila using the ultracentrifugation method and denoted as AhEVs. Using transmission electron microscopy imaging, we confirmed the ultrastructure and spherical shape morphology of AhEVs. Nanoparticle-tracking analysis results showed a mean particle size of 105.5 ± 2.0 nm for AhEVs. Moreover, the particle concentration of AhEVs was 2.34 ± 0.12 × 1011 particles/mL of bacterial supernatant. AhEV-treated fathead minnow (FHM) cells did not show cytotoxicity effects up to 50 µg/mL with no significant decrease in cells. Moreover, no mortality was observed in larval zebrafish up to 50 µg/mL which indicates that the AhEVs are biocompatible at this concentration. Furthermore, fluorescent-labeled AhEVs were internalized into FHM cells. Results of qRT-PCR analysis in FHM cells revealed that cellular pro-inflammatory cytokines such as nuclear factor (NF)-κB, interferon (Ifn), Irf7, interleukin (Il) 8, and Il11 were upregulated while downregulating the expression of anti-inflammatory Il10 in a concentration-dependent manner. AhEV-treated adult zebrafish (5 µg/fish) induced toll-like receptor (tlr) 2 and tlr4; tumor necrosis factor-alpha (tnfα); heat shock protein (hsp) 70; and il10, il6, and il1ß in kidney. Protein expression of NF-κB p65 and Tnfα presented amplified levels in the spleen of AhEVs-treated zebrafish. Based on the collective findings, we conclude that AhEVs exhibited morphological and physicochemical characteristics to known EVs of gram (-)ve bacteria. At biocompatible concentrations, the immunomodulatory activity of AhEVs was demonstrated by inducing different immune response genes in FHM cells and zebrafish. Hence, we suggest that AhEVs could be a novel vaccine candidate in fish medicine due to their ability to elicit strong immune responses.
Asunto(s)
Aeromonas hydrophila , Vesículas Extracelulares , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Pez Cebra , Animales , Aeromonas hydrophila/fisiología , Vesículas Extracelulares/inmunología , Vesículas Extracelulares/química , Enfermedades de los Peces/inmunología , Pez Cebra/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Bacterias Gramnegativas/inmunología , Cyprinidae/inmunología , Factores Inmunológicos/farmacología , Factores Inmunológicos/química , Línea Celular , Agentes Inmunomoduladores/química , Agentes Inmunomoduladores/farmacologíaRESUMEN
Bacterial septicemia in freshwater fish is mainly caused by Aeromonas hydrophila infection, which affects the development of aquaculture industry. In the context of sustainable aquaculture, subunit vaccines are of great values because they play positive roles in reducing the overuse of antibiotics and protecting aquatic animals against bacterial infection. In this study, the recombinant outer membrane protein OmpTS of A. hydrophila were used as subunit vaccine to immunize Megalobrama amblycephala, and its immunoprotective effect and host immune responses were evaluated. The survival rates of the vaccinated groups after bacterial infection were significantly higher than that of the control group, especially of the OmpTS high-dose vaccinated group. The better protective effects of vaccinated groups might be attributed to the increased levels of serum IgM-specific antibody titer, the reduced relative abundance of A. hydrophila in various tissues, the increased number of immune-positive cells with different epitopes, the up-regulated expression levels of immune-related genes, and the enhanced activities of antibacterial enzymes. In conclusion, OmpTS subunit vaccine could strongly induce host immune responses in M. amblycephala, thereby enhancing both cellular and humoral immunity, which exhibited excellent and effective immunoprotective efficacy.
Asunto(s)
Aeromonas hydrophila , Vacunas Bacterianas , Cyprinidae , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Vacunas de Subunidad , Aeromonas hydrophila/inmunología , Animales , Enfermedades de los Peces/prevención & control , Enfermedades de los Peces/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/prevención & control , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/administración & dosificación , Vacunas de Subunidad/inmunología , Vacunas de Subunidad/administración & dosificación , Cyprinidae/inmunología , Proteínas de la Membrana Bacteriana Externa/inmunología , Inmunidad HumoralRESUMEN
This study marks the first utilization of reverse vaccinology to develop recombinant subunit vaccines against Pseudomonas koreensis infection in Empurau (Tor tambroides). The proteome (5538 proteins) was screened against various filters to prioritize proteins based on features that are associated with virulence, subcellular localization, transmembrane helical structure, antigenicity, essentiality, non-homology with the host proteome, molecular weight, and stability, which led to the identification of eight potential vaccine candidates. These potential vaccine candidates were cloned and expressed, with six achieving successful expression and purification. The antigens were formulated into two distinct vaccine mixtures, Vac A and Vac B, and their protective efficacy was assessed through in vivo challenge experiments. Vac A and Vac B demonstrated high protective efficacies of 100 % and 81.2 %, respectively. Histological analyses revealed reduced tissue damage in vaccinated fish after experimental infection, with Vac A showing no adverse effects, whereas Vac B exhibited mild degenerative changes. Quantitative real-time PCR results showed a significant upregulation of TNF-α and downregulation of IL-1ß in the kidneys, spleen, gills, and intestine in both Vac A- and Vac B-immunized fish after challenged with P. koreensis. Additionally, IL-8 exhibits tissue-specific differential expression, with significant upregulation in the kidney, gills, and intestine, and downregulation in the spleen, particularly notable in Vac A-immunized fish. The research underscores the effectiveness of the reverse vaccinology approach in fish and demonstrates the promising potential of Vac A and Vac B as recombinant subunit vaccines.
Asunto(s)
Enfermedades de los Peces , Infecciones por Pseudomonas , Pseudomonas , Animales , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/prevención & control , Pseudomonas/inmunología , Infecciones por Pseudomonas/veterinaria , Infecciones por Pseudomonas/prevención & control , Infecciones por Pseudomonas/inmunología , Vacunas de Subunidad/inmunología , Vacunas de Subunidad/administración & dosificación , Vacunas Bacterianas/inmunología , Vacunas Bacterianas/administración & dosificación , Vacunología , Vacunas Sintéticas/inmunología , Cyprinidae/inmunología , Vacunas contra la Infección por Pseudomonas/inmunología , Proteoma/inmunologíaRESUMEN
Interleukin-10 (IL-10) is an immunosuppressive cytokine, which plays a vital role in regulating inflammation for inhibiting the generation and function of pro-inflammatory cytokines in vivo or in vitro. In the present study, the full length cDNA of IL-10 was characterized from Nibea albiflora (named as NaIL-10) of 1238 base pairs (bp), containing a 5'-UTR (untranslated region) of 350 bp, a 3'-UTR of 333 bp and an open reading frame (ORF) of 555 bp (Fig. 1A) to encode 184 amino acid residues with a signal peptide at the N-terminus. The sequence analysis showed that NaIL-10 possessed the typical IL-10 family symbolic motif and conversed cysteine residues, similar to its teleost orthologues. Real-time PCR indicated that NaIL-10 had wide distribution in different healthy tissues, with a relatively high expression in immune-related tissues (head kidney, spleen, kidney, liver and gill). Significantly, up-regulations of NaIL-10 after infection against Vibrio parahaemolyticus, Vibrio alginolyticus and Poly I:C were also observed. Subcellular localization manifested that NaIL-10 mainly distributed in the cytoplasm unevenly and aggregately, and there was also a small amount on the cell membrane, indicating that NaIL-10 was secreted to the extracellular space as the known IL-10 homologous molecules. It could co-locate with IL-10 Rα on the membrane of HEK293T cells for their potential interaction, and GST pull-down and Co-IP studies certified the specific and direct interaction between NaIL-10 and NaIL-10 Rα, confirming that an IL-10 ligand-receptor system existed in N.albiflora. The expression of pro-inflammatory cytokines, including TNF-α, IL-6, IL-1ß, were dramatically inhibited in LPS-stimulated RAW264.7 macrophages pre-incubated with recombinant NaIL-10 protein, demonstrating its anti-inflammatory roles. Taken together, the results demonstrated the existence of IL-10 ligand-receptor system in N.albiflora for the first time, and indicated the suppressive function of NaIL-10 on pro-inflammatory cytokine expression in inflammatory response, which would be conducive to better comprehending the role of IL-10 in the immunomodulatory mechanisms of teleost.
Asunto(s)
Secuencia de Aminoácidos , Enfermedades de los Peces , Proteínas de Peces , Regulación de la Expresión Génica , Inmunidad Innata , Interleucina-10 , Filogenia , Animales , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Proteínas de Peces/química , Enfermedades de los Peces/inmunología , Interleucina-10/genética , Interleucina-10/inmunología , Inmunidad Innata/genética , Regulación de la Expresión Génica/inmunología , Regulación de la Expresión Génica/efectos de los fármacos , Vibrio parahaemolyticus/fisiología , Alineación de Secuencia/veterinaria , Perfilación de la Expresión Génica/veterinaria , Poli I-C/farmacología , Vibriosis/inmunología , Vibriosis/veterinaria , Cyprinidae/inmunología , Cyprinidae/genética , Vibrio alginolyticus/fisiología , Secuencia de BasesRESUMEN
The present study explores the use of periphyton to ameliorate toxic properties of arsenic (As) to Labeo rohita and also assesses the human food safety aspects. Fish were introduced to arsenite [As(III)] contaminated water (0.3 and 3 mg/L) along with periphyton. Biochemical, physiological and immunological parameters, including gene expression, were assessed after 30 days of exposure. Periphyton incorporation significantly improved (p < 0.05) the adverse effects of As on respiration, NH3 excretion and brain AChE activity by reducing oxidative stress and As bioaccumulation. The presence of periphyton in As(III) exposed fish (3 mg/L) increased the immune response (Immunoglobulin M and Complement C3) in the serum and the regulation of the respective immune genes in the anterior kidney was found to be similar to the control. A speciation study using LC-ICP-MS confirmed the high accumulation of As by periphyton (5.0-31.9 µg/g) as arsenate [As (V)], resulting in a lower amount of As in fish muscle. The calculated human health risk indices, Target Hazard Quotient (THQ) and Target Cancer risk (TCR) indicate that fish grown in periphyton-treated water may lower the human health risks associated with As. The study signifies the importance of periphyton-based aquaculture systems in As contaminated regions for safe fish production with enhanced yield.
Asunto(s)
Arsénico , Bioacumulación , Cyprinidae , Estrés Oxidativo , Contaminantes Químicos del Agua , Animales , Contaminantes Químicos del Agua/toxicidad , Medición de Riesgo , Cyprinidae/inmunología , PecesRESUMEN
Culter alburnus is sensitive to stressors. Arginine is a precursor of nitric oxide, which can effectively relieve the level of oxidative stress and improve the antioxidant and immune capacity of fish. The effect of different arginine levels on topmouth culter (Culter alburnus) fry development performance, liver antioxidant capacity, and immune parameters were investigated in this study. Five diets (1.96%, ARG1, control group; 2.28%, ARG2; 2.52%, ARG3; 2.81%, ARG4; 3.09%, ARG5) were used to feed fry (initial weight 0.31 ± 0.01 g) for 8 weeks. The data showed that the final weight (FW), weight gain rate (WGR), and specific growth rate (SGR) of the ARG3 and ARG4 groups were significantly improved, while the feed conversion ratio (FCR) reduced significantly. Compared with the ARG1 group, all groups remarkably reduced the crude ash content of the whole body. The activity of hepatic superoxide dismutase (SOD) and the content of hepatic glutathione (GSH) were significantly increased in the ARG3 and ARG4 groups, while the malondialdehyde (MDA) content was significantly decreased. Compared with the ARG1 group, arginine levels in ARG2, ARG3, and ARG4 groups up-regulated the expression levels of Nrf2, down-regulated the gene expression level of Keap1 in the liver. And the expression of Nrf2/Keap1 pathway downstream genes Mn-SOD and CAT was up-regulated in ARG2 and ARG3 groups. Furthermore, the expression levels of MyD88 and IL-1ß were down-regulated, and the anti-inflammatory gene TGF-ß expression levels were up-regulated in the ARG2, ARG3, and ARG4 groups. Additionally, compared to the ARG1 group, there was a significant increase in the relative expression levels of the C3 and C4 genes in the ARG4 group. In conclusion, 2.28-2.81% dietary arginine levels improved the growth performance, promoted antioxidant capacity, and enhance immune response. The optimal level of arginine was determined by the quadratic regression analysis of SGR and FCR to be 2.55% of diet (5.43% of dietary protein) and 2.53% of diet (5.38% of dietary protein), accordingly.
Asunto(s)
Alimentación Animal , Antioxidantes , Arginina , Cyprinidae , Dieta , Animales , Arginina/administración & dosificación , Arginina/farmacología , Antioxidantes/metabolismo , Dieta/veterinaria , Alimentación Animal/análisis , Cyprinidae/crecimiento & desarrollo , Cyprinidae/inmunología , Hígado/metabolismo , Suplementos Dietéticos , Proteínas de Peces/metabolismo , Proteínas de Peces/genéticaRESUMEN
This study was designed to determine the effects of papaya peel extract (PPE) supplementation on the growth and immunophysiological responses of rohu fingerlings at different stocking densities. In this study, three isonitrogenous (307.2-309.8 g kg-1 protein) and isocaloric diets (16.10-16.16 MJ digestible energy kg-1) were prepared using three different inclusion levels (0, 5, and 10 g kg-1) of PPE. Four hundred and five rohu fingerlings (mean weight: 4.24 g ± 0.12) were randomly distributed into nine treatment groups in triplicates viz. low (10nos 75 L-1 or ≈ 0.565 kg/m3), medium (15nos 75 L-1 or ≈ 0.848 kg/m3), and high (20nos 75 L-1 or ≈ 1.13 kg/m3) following a completely randomized design. The study found that increasing stocking density negatively affected fish growth indices, such as weight gain percentage (WG%), feed efficiency ratio (FER), specific growth rate (SGR) and survival. In contrast, dietary PPE supplementation improved growth indices and survival (p < 0.05). We also observed that aminotransferase, lactate (LDH), and malate dehydrogenase (MDH) activity increased with stocking density, whereas 5 and 10 g kg-1 PPE supplementation reduced LDH and MDH activity (p < 0.05). PPE supplementation positively affected serum indices, decreased glucose levels, and increased respiratory burst activity (p < 0.05). Interferon-gamma (IFN-γ) expression was highest in the low- and medium-stocking density groups fed with 5 g kg-1 PPE, which also increased total immunoglobulin and myeloperoxidase activity while decreasing malondialdehyde concentration (p < 0.05). The results revealed that 5 g kg-1 dietary PPE supplementation could be used as a growth promoter and immunostimulant to improve immuno-physiological responses at low and medium stocking densities.
Asunto(s)
Alimentación Animal , Carica , Cyprinidae , Dieta , Suplementos Dietéticos , Extractos Vegetales , Animales , Carica/química , Extractos Vegetales/farmacología , Extractos Vegetales/administración & dosificación , Alimentación Animal/análisis , Dieta/veterinaria , Cyprinidae/inmunología , Cyprinidae/crecimiento & desarrollo , Aglomeración , Estrés Fisiológico/efectos de los fármacosRESUMEN
A study was conducted to evaluate the effect of Piper nigrum (black pepper) leaf extract on on the growth performance, proximate composition, hematological parameters, and immune response of Labeo rohita fingerlings with an average weight of 22.14 ± 0.98g. After acclimation for two weeks, fish (n=25) were randomly selected and placed in four glass aquaria (T0, T1, T2 and T3) at constant water temperature (30.0 ± 1.0 °C), pH (7.50 ± 0.5) and total hardness (200 ± 2.0 mgL,-¹) for a period of 12 weeks, with three replicates each. Fish were fed with P. nigrum leaf extract supplemented feed @ 0.0%, 1.0%, 2.0% and 3.0% in T0, T1, T2 and T3, respectively. At the end of experiment, five fish were randomly selected from each aquaria for proximate composition, gut and skin microbial load, hematological parameters. Total proteins, albumins, and globulins were also recorded to evaluate immunological memory. The result revealed that fish in T2 showed better growth performance with an average weight gain of 56.11 ± 0.51 g. Thus, it had been concluded that Piper nigrum, a medicinal plant, can also be used to enhance the growth performance and immune response of Labeo rohita as attractive alternatives against antibiotics and vaccines and has shown no negative side effects on fish health as well as on its environment.
Um estudo foi conduzido para avaliar o efeito do extrato da folha de Piper nigrum (pimenta-do-reino) sobre o desempenho de crescimento, composição centesimal, parâmetros hematológicos e resposta imune de alevinos de Labeo rohita com peso médio de 22,14 ± 0,98g. Após aclimatação por duas semanas, os peixes (n = 25) foram selecionados aleatoriamente e colocados em quatro aquários de vidro (T0, T1, T2 e T3) em temperatura constante da água (30,0 ± 1,0 °C), pH (7,50 ± 0,5) e dureza total (200 ± 2,0 mgL-¹) por um período de 12 semanas, com três repetições cada. Os peixes foram alimentados com ração suplementada com extrato de folha de P. nigrum @ 0,0%, 1,0%, 2,0% e 3,0% em T0, T1, T2 e T3, respectivamente. Ao final do experimento, cinco peixes foram selecionados aleatoriamente de cada aquário para composição centesimal, carga microbiana intestinal e cutânea e parâmetros hematológicos. Proteínas totais, albuminas e globulinas também foram registradas para avaliar a memória imunológica. O resultado revelou que os peixes em T2 apresentaram melhor desempenho de crescimento com ganho de peso médio de 56,11 ± 0,51 g. Assim, concluiu-se que Piper nigrum, uma planta medicinal, também pode ser usado para melhorar o desempenho de crescimento e resposta imunológica de Labeo rohita como alternativas atraentes contra antibióticos e vacinas e não mostrou efeitos colaterais negativos na saúde dos peixes, bem como sobre seu ambiente.
Asunto(s)
Animales , Cyprinidae/crecimiento & desarrollo , Cyprinidae/inmunología , Cyprinidae/sangre , Piper/químicaRESUMEN
The effect of four level of Astragalus polysaccharides (APs) supplementation diets, (CD: control diet and three experiment diet (E), EA: 100 mg kg-1 APs; EB: 200 mg kg-1 APs; EC: 300 mg kg-1 APs) on growth, changes in haemato-biochemical parameters and metabolic-digestive enzymes, enhancement of antioxidant activity, innate-adaptive immune response, and cytokine gene expression were studied in catla (Catla catla) against Edwardsiella tarda. The healthy and challenged groups fed the CD displayed no mortality, while fish fed EA or EC revealed 10% mortality, but the mortality was only 5% in diet EB. Fish fed diet EB and EC revealed significantly better growth rates and high RBC count during the experimental period. Albumin and globulin levels were significant improved when fish were fed the diet EB and EC from weeks 6-8. The superoxide dismutase (SOD) was significant ameliorated by EB feeding from weeks 4-8. In contrast, serum myeloperoxidase (MPO), catalase (CAT), malondialdehyde (MDA)/lipid peroxidation (LPO), glutathione peroxidase (GPx), respiratory burst activity (RBA), bactericidal action (BCA), serum lysozyme activity (SLA), nitric oxide synthase (NOS), head kidney leukocytes response proliferation (HKLP), hemolytic action (HLA), hydrogen peroxides (H2O2), and immunoglobulin (Ig) were significantly improved from week 6-8. Groups fed the APs enriched diets had significant ameliorated interleukin (IL)-1ß and interferon (IFN)-γ mRNA expression after 6 and 8 weeks of feeding. However, IL-10 and major histocompatibility complex (MHC)-1 mRNA expressions were significant enhanced in catla fed all APs diets on week 8. APs enriched diets revealed significant improved tumor necrosis factor (TNF)-α and TNF receptor-associated factor-6 (TRAF6) mRNA expression on week 4, but toll-like receptor-2 (TLR2) and TLR4 mRNA expression were significant enhanced by diet EB and EC after weeks 6 and 8. Similarly, the lysozyme (Lyz)-C and Lyz-G mRNA levels in the head kidney (HK) increased by APs feeding on weeks 6 and 8, whereas the EB diet, the expression of nucleotide binding oligomerization domain-1 (NOD1) was significantly improved on weeks 6 and 8, but NOD2 mRNA expression was only significant enhanced after 8 weeks of diet EB. By feeding healthy catla and E. tarda challenged fish fed diet EB, resulted in significantly increased growth, haemato-biochemical indices, metabolic-digestive enzymes, antioxidant activities, innate-adaptive immune responses, and cytokine gene expression mainly between 6 and 8 weeks.
Asunto(s)
Cyprinidae , Dieta , Infecciones por Enterobacteriaceae/veterinaria , Enfermedades de los Peces , Polisacáridos/administración & dosificación , Alimentación Animal/análisis , Animales , Antioxidantes/metabolismo , Planta del Astrágalo/química , Cyprinidae/crecimiento & desarrollo , Cyprinidae/inmunología , Cyprinidae/microbiología , Citocinas , Dieta/veterinaria , Suplementos Dietéticos , Edwardsiella tarda/patogenicidad , Infecciones por Enterobacteriaceae/inmunología , Peróxido de Hidrógeno , Inmunidad , Muramidasa , ARN MensajeroRESUMEN
The present study investigates the effects of Supplementary diet Hygrophila auriculata on the growth, survival, biochemical and haematological parameters of Cirrhinus mrigala. The seaweed was administered to the fish possessing an initial average weight of 14.063 ± 1.828 g. Fish were fed with supplementary diet H. auriculata exhibited significant difference (P < 0.05) in the growth performance, haematological indices such as RBC count, haematocrit volume, haemoglobin, WBC, MCV, MCH and MCHC concentration in contrast to the control after a period of 8 weeks. Also, there were significant differences in biochemical parameters (P < 0.05), between the fish supplemented with dietary H. auriculata extract and the control group. These findings suggest that the administration of H. auriculata extract has a positive effect on the immunological indices and the immune system activity in Mrigal fish.
Asunto(s)
Alimentación Animal , Cyprinidae/sangre , Cyprinidae/crecimiento & desarrollo , Dieta , Acanthaceae , Animales , Acuicultura , Cyprinidae/inmunología , Suplementos Dietéticos , Agua Dulce , Plantas Medicinales , Algas MarinasRESUMEN
Nutritional programming signifies a process in which broodstock feeding approaches have long-term effects on the subsequent progeny. The present study aimed to elucidate whether supplementing golden mahseer, Tor putitora broodstock diets with ß-glucan affects progeny growth performance, survival, thermal tolerance, and non-specific immunity. Initially, the growth performance of progeny produced from brooders fed with different levels of ß-glucan was non-significant. However, on the 15th and 35th DPH, the maximum weight was observed in fry obtained from the brooders fed with 0.5% followed by 1.0% ß-glucan. Furthermore, on 50th DPH, significantly higher weight was registered in the fry from the 0.5% ß-glucan fed group while 1.0% ß-glucan group had no transgenerational effect on growth. The condition factor of fry obtained from golden mahseer brooders fed with a 0.5% ß-glucan diet was greater than the control and 1.0% ß-glucan fed group. On the other hand, we did not find any significant transgenerational influence of ß-glucan on the survival of the progeny. The thermal tolerance of fry produced from brooders fed with ß-glucan was significantly modulated at both end-points (CTmax and CTmin). Expression of interleukin-1ß was significantly up-regulated in fry obtained from ß-glucan fed brooders. In contrast, the expression level of tumor necrosis factor-α was significantly higher only in fry produced from 1.0% ß-glucan fed brooders. The expression of immunoglobulin light chain and serum amyloid A gene was significantly higher in fry produced from 0.5% ß-glucan fed brooders. Overall results suggest that the dietary provisioning of ß-glucan in golden mahseer brooders can be a strategy to produce healthy and robust fry in captivity for stock enhancement and conservation programs.
Asunto(s)
Cyprinidae/crecimiento & desarrollo , Especies en Peligro de Extinción , Termotolerancia/efectos de los fármacos , beta-Glucanos/farmacología , Animales , Cyprinidae/inmunología , Cyprinidae/metabolismo , Suplementos Dietéticos , Femenino , Larva/efectos de los fármacos , MasculinoRESUMEN
Experiment was conducted to assess the impact of ginger (Zingiber officinale) as a dietary supplement with probiotic bacterium Bacillus coagulans on growth performance, hematological parameters, and non-specific immune parameters in Catla catla. An attempt was also made to investigate their effects on histology and extent of DNA damage against pathogenic bacterium Aeromonas hydrophila in C. catla. Ten dietary treatments were designed in triplicate in which five groups of fingerlings fed on diets containing different incorporation level of dried ginger at 0 g kg-1 (C1; basal diet 1 with duckweed as major protein source), 1 g kg-1 (G1), 5 g kg-1 (G2), 10 g kg-1 (G3), and 15 g kg-1 (G4) and other five groups of fingerlings fed on diets (G5-G9) containing different incorporation level of dried ginger at 0, 1, 5, 10, and 15 g kg-1 along with probiotic bacterium B. coagulans @ 3000 CFU ml-1 for 90 days. Results clearly revealed significantly (P < 0.05) high values of weight gain, growth % gain in body weight, specific growth rate (SGR), RBC and WBC count, phagocytic activity, respiratory burst activity, serum protein, and serum bactericidal activity in group of fingerlings fed on ginger supplemented diets which further improved with the inclusion of probiotic with best results in treatment G8. Histopathological study and COMET assay reflected that Z. officinale and probiotics protect extent of DNA damage and the digestive organs from the detrimental effects of pathogenic bacteria affirming their positive role and harmonious effects of probiotic bacterium with Z. officinale-supplemented diets.
Asunto(s)
Bacillus coagulans , Cyprinidae , Dieta , Probióticos , Zingiber officinale , Alimentación Animal/análisis , Animales , Cyprinidae/crecimiento & desarrollo , Cyprinidae/inmunología , Daño del ADN , Dieta/veterinaria , Zingiber officinale/química , Inmunización , Probióticos/farmacologíaRESUMEN
Chitooligosaccharide (COS) is an important immune enhancer and has been proven to have a variety of biological activities. Our previous research has established an M1 polarization mode by COS in blunt snout bream (Megalobrama amblycephala) macrophages, but the mechanism of COS activation of blunt snout bream macrophages remains unclear. In this study, we further explored the internalization mechanism and signal transduction pathway of chitooligosaccharide hexamer (COS6) in blunt snout bream macrophages. The results showed that mannose receptor C-type lectin-like domain 4-8 of M. amblycephala (MaMR CTLD4-8) could recognize and bind to COS6 and mediate COS6 into macrophages by both clathrin-dependent and caveolin-dependent pathways. In the inflammatory response of macrophages activated by COS6, the gene expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and nitric oxide synthase 2 (NOS2) was significantly inhibited after MaMR CTLD4-8-specific antibody blockade. However, even if it was blocked, the expression of these inflammation-related genes was still relatively upregulated, which suggested that there are other receptors involved in immune regulation. Further studies indicated that MaMR CTLD4-8 and Toll-like receptor 4 (TLR4) cooperated to regulate the pro-inflammatory response of macrophages caused by COS6. Taken together, these results revealed that mannose receptor (MR) CTLD4-8 is indispensable in the process of recognition, binding, internalization, and immunoregulation of COS in macrophages of blunt snout bream.
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Quitosano/metabolismo , Cyprinidae/inmunología , Expresión Génica , Lectinas Tipo C/metabolismo , Macrófagos/inmunología , Lectinas de Unión a Manosa/metabolismo , Oligosacáridos/metabolismo , Receptores de Superficie Celular/metabolismo , Animales , Anticuerpos/efectos adversos , Cyprinidae/metabolismo , Interleucina-1beta/metabolismo , Receptor de Manosa , Óxido Nítrico Sintasa de Tipo II/metabolismo , Transducción de Señal , Linfocitos T Reguladores/inmunología , Receptores Toll-Like/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
p65 is one of the important subunits of the inflammation-related transcription factor NF-κB. In the present study, we cloned and identified the p65 from Megalobrama amblycephala (Mnp65) by homologous cloning and RACE technique. The full-length Mnp65 cDNA consisted of 2331 bp, and included one open reading frame encoding a 604-amino acid putative protein. The protein sequence included a DNA binding motif, a well conserved N-terminal Rel-homology domain (RHD), and a C-terminal IG-like plexins transcription (IPT). Mnp65 was closely related with the other p65 proteins of Cypriniformes and clearly distinct from that of Perciformes and Salmoniformes in terms of sequence homology. Mnp65 homodimer may interact with IκBα in the IPT domain based on the predicted 3D structure of IκBα/Mnp65 complex. Mnp65 was ubiquitously expressed in M. amblycephala tissues, and the highest levels were detected in muscle and liver. Intragastric infection with Aeromonas hydrophila caused respiratory burst and cytokine storm from 8 h to 48 h, showing significantly higher level of respiratory burst activities and significantly high cytokines levels, such as TNF-α, IL-1ß, IL-6, IL-8 etc., compared to 0 h. In addition, the bacterial challenge downregulated the IkBα, and upregulated Mnp65 and TNF-α in the liver. IkBα-Mnp65 was regulated by the negative feedback of cytokine storm, to increase IkBα and decrease Mnp65. Then cytokine storm was relieved at 96 h. Finally, severe intestinal inflammation was observed from 24 h to 48 h after infection, characterized by extensive villous necrosis, epithelial hyperplasia and lymphocyte infiltration, all of which were relieved at 96 h. Taken together, Mnp65 plays a crucial role in the physiological response of teleost fish to bacterial infection.
Asunto(s)
Aeromonas hydrophila/metabolismo , Cyprinidae/microbiología , Síndrome de Liberación de Citoquinas/inmunología , Enfermedades de los Peces/inmunología , Proteínas de Peces/metabolismo , Inflamación/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Cyprinidae/genética , Cyprinidae/inmunología , Cyprinidae/metabolismo , Síndrome de Liberación de Citoquinas/metabolismo , Síndrome de Liberación de Citoquinas/microbiología , Síndrome de Liberación de Citoquinas/patología , Enfermedades de los Peces/metabolismo , Enfermedades de los Peces/microbiología , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Inflamación/metabolismo , Inflamación/microbiología , Inflamación/patología , Filogenia , Conformación Proteica , Estallido RespiratorioRESUMEN
This study reports the effect of ulvan enriched diet on the influence of growth, changes in hemato-biochemical indices, improvement of antioxidant system, enhancement of innate-adaptive immunity and modification of immuno-antioxidant genes expression in Labeo rohita against Flavobacterium columnaris. The weight gain (WG) was significantly high (P > 0.05) in unchallenged normal and challenged fish fed with diets enriched with 25 and 50 mg kg-1 ulvan; the FCR was better (P > 0.05) when fed with 50 mg kg-1 enriched diet. In normal fish fed with or without ulvan supplementation was noted 100% survival rate (SR). In both groups, the red blood cell (RBC) and while blood cell (WBC) counts increased significantly (P > 0.05) when fed with 50 mg kg-1 ulvan diet whereas the hemoglobin (Hb) level increased significantly on being fed with 25 and 50 mg kg-1 ulvan diets. The SOD activity was enhanced significantly in both groups fed with any dose of ulvan diets whereas the MDA and GPx activity increased only with 25 and 50 mg kg-1 ulvan diets. The phagocytic (PC) activity significantly increased with any enriched diet and control diet groups while the respiratory burst (RB) activity increased only with 50 mg kg-1 ulvan diet. The alternate complement pathway (ACP), activity of lysozyme (Lyz), and immunoglobuline M (IgM) were better in both groups fed with 50 mg kg-1 ulvan diet. The SOD and GPx antioxidant gene expression were significantly high in both groups fed with any ulvan diet while the Nrf2 gene expression was high with 50 mg kg-1 ulvan diet. The IL-1ß, TNFα, hepcidin, Lyz, and IgM cytokines or proteins mRNA expression were significant in both groups fed with all ulvan supplement diet whereas the ß-2M expression was significant only with 50 mg kg-1 ulvan diet. The present research indicates that both L. rohita groups fed with 50 mg kg-1 ulvan diet significantly improved growth, antioxidant system, immune defense system, and immuno-antioxidant related gene expression against F. columnaris.
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Cyprinidae , Enfermedades de los Peces , Infecciones por Flavobacteriaceae , Flavobacterium , Factores Inmunológicos/farmacología , Polisacáridos/farmacología , Animales , Cyprinidae/genética , Cyprinidae/crecimiento & desarrollo , Cyprinidae/inmunología , Cyprinidae/microbiología , Enfermedades de los Peces/sangre , Enfermedades de los Peces/genética , Enfermedades de los Peces/inmunología , Infecciones por Flavobacteriaceae/sangre , Infecciones por Flavobacteriaceae/genética , Infecciones por Flavobacteriaceae/inmunología , Infecciones por Flavobacteriaceae/veterinaria , Regulación de la Expresión Génica/efectos de los fármacos , Glutatión/inmunología , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/inmunología , Riñón Cefálico/efectos de los fármacos , Riñón Cefálico/inmunología , Inmunoglobulina M/sangre , Inmunoglobulina M/genética , Malondialdehído/inmunología , Muramidasa/sangre , Muramidasa/genética , Superóxido Dismutasa/genética , Superóxido Dismutasa/inmunología , Aumento de Peso/efectos de los fármacosRESUMEN
Current evidence suggests thyroid hormones (THs) impact development of the immune system, but few studies have explored the connection between the thyroid and immune systems, especially in fish. This is important as some environmental contaminants disrupt TH homeostasis and may thus have negative impacts on the immune system. To determine the long-term consequences of early life stage (ELS) hypothyroidism on immune function, fathead minnows were exposed to the model thyroid hormone suppressant propylthiouracil (PTU) from < 1 to 30 days post hatch. Fish were transferred to clean water and raised to adulthood (5-7 months post hatch) at which time, several aspects of immune function were evaluated. Ex vivo assessment of immune cell function revealed significant decreases (1.2-fold) in the phagocytic cell activity of PTU-treated fish relative to the controls. Fish were also injected with Yersinia ruckeri to evaluate their in vivo immune responses across a suite of endpoints (i.e., transcriptomic analysis, leukocyte counts, spleen index, hematocrit, bacterial load and pathogen resistance). The transcriptomic response to infection was significantly different between control and PTU-treated fish, though no differences in bacterial load or pathogen resistance were noted. Overall, these results suggest that early life stage TH suppression causes long-term impacts on immune function at the molecular and cellular levels suggesting a key role for TH signaling in normal immune system development. This study lays the foundation for further exploration into thyroid-immune crosstalk in fish. This is noteworthy as disruption of the thyroid system during development, which can occur in response to chemicals present in the environment, may have lasting effects on immune function in adulthood.
Asunto(s)
Cyprinidae/inmunología , Enfermedades de los Peces/inmunología , Interacciones Huésped-Patógeno/inmunología , Glándula Tiroides/fisiopatología , Yersiniosis/veterinaria , Animales , Antitiroideos/farmacología , Femenino , Enfermedades de los Peces/microbiología , Proteínas de Peces/genética , Regulación de la Expresión Génica , Interacciones Huésped-Patógeno/fisiología , Inmunidad Celular , Inmunidad Innata , Riñón/microbiología , Riñón/fisiología , Propiltiouracilo/farmacología , Glándula Tiroides/efectos de los fármacos , Glándula Tiroides/inmunología , Hormonas Tiroideas/metabolismo , Yersiniosis/inmunología , Yersiniosis/microbiología , Yersinia ruckeriRESUMEN
To understand the intestinal microbial diversity and community structure of bighead carp (Aristichthys nobilis) under different feeding strategies, 39 fish from three groups (A: 9 fish, natural live food only; B: 15 fish, natural live food + fish formulated feeds; C: 15 fish, natural live food + fish formulated feed + lactic acid bacteria) were obtained for the high throughput 16S rRNA gene sequencing. We first examined five non-specific immunity indications of the carp-lysozyme (LZM), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GSH-PX), and superoxide dismutase (SOD). Interestingly, the composition of gut microbiota and related non-specific immune indices were affected by the feeding treatment of the bighead carp. Notably, all enzyme activity indexes were significantly different (p < 0.01) in the spleen and three enzyme activity indexes (LZM, GSH-PX, and SOD) had significant differences in the hepatopancreas (p < 0.001) of the carp from the three groups. The 16S rRNA gene sequencing showed higher diversity in groups B and C. Compared to group A, the relative abundance of Actinobacteria increased significantly and the relative abundance of Proteobacteria and Firmicutes decreased significantly in groups B and C at the phylum level. Functional analysis revealed the association between non-specific immune indicators and import genera in the hepatopancreas and spleen of bighead carp. This study provides new insights into the gut microbiomes and non-specific immune of bighead carp.