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1.
Mol Immunol ; 125: 83-94, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32652363

RESUMO

Leukocyte immune-type receptors (LITRs) are a multigene family of teleost immunoregulatory proteins that share structural, phylogenetic, and likely functional relationships with several innate immune receptor proteins in other vertebrates, including mammals. Originally discovered in channel catfish (Ictalurus punctatus), representative IpLITR-types have been shown to regulate diverse innate immune cell effector responses including phagocytosis, degranulation, and cytokine secretion. To date, IpLITRs have been primarily characterized using mammalian cell line expression systems, therefore many unanswered questions remain regarding their actual regulatory roles in fish immunity. In the present study, we report on the preliminary molecular characterization of five goldfish (Carassius auratus) CaLITR-types and the identification of several putative splice variants of these receptors cloned from various goldfish tissues and primary myeloid cell cultures. In general, CaLITR mRNA transcripts were detected in all goldfish tissues tested, and also in primary kidney macrophage and neutrophil cultures. Specifically, CaLITR1 is a functionally ambiguous receptor with no charged amino acids in its transmembrane (TM) segment and is devoid of tyrosine-based signaling motifs in its short cytoplasmic tail (CYT) region. CaLITR2 is a putative activating receptor-type that contains immunotyrosine-based activation motifs (ITAMs) within its long CYT region, and CaLITR3 has a positively charged TM segment, suggesting that it may recruit intracellular stimulatory adaptor signaling molecules. CaLITR4 and CaLITR5 appear to have diverse signaling capabilities since they contain various immunoregulatory signaling motifs within their CYT regions including putative Nck and STAT recruitment motifs as well as ITAM-like and ITIM sequences. We also identified putative CaLITR splice variants with altered extracellular Ig-like domain compositions and variable CYT regions. Interestingly, this suggests that alternative splicing-mediated diversification of CaLITRs can generate receptor forms with possible variable binding and/or intracellular signaling abilities. Overall, these findings reveal new information about the teleost LITRs and sets the stage for exploring how alternative splicing leads to the functional diversification of this complex multigene immunoregulatory receptor family.


Assuntos
Carpa Dourada/imunologia , Imunidade Inata/imunologia , Leucócitos/imunologia , Receptores Imunológicos/genética , Receptores Imunológicos/imunologia , Processamento Alternativo , Animais , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Carpa Dourada/genética , Imunidade Inata/genética
2.
Fish Shellfish Immunol ; 98: 285-295, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31962149

RESUMO

As one of the most important fish in freshwater aquaculture, gibel carp (Carassius auratus gibelio) is easily susceptible to Cyprinid herpesvirus 2 (CyHV-2). Immersion vaccination has attracted many researchers due to its simple operation in preventing infectious diseases. However, the unavoidable disadvantage is that the immersion vaccine must be used with adjuvants to get a better performance. In this study, gibel carps were vaccinated by a 60 min bath in a ß-propiolactone-inactivated Cyprinid herpesvirus 2, mixed with DTT, ß-glucan, anisodamine and scopolamine, respectively. After immunization, the fishs were challenged by CyHV-2 in 2 weeks. By analyzing pathological section, we found that ß-glucan, anisodamine and scopolamine groups protected the gibel carp compared to the control group, which was consistent with the trend of survival rate. Specifically, ß-glucan group in serum appeared best on lysozyme, TSOD and complement C3. Real time quantitative RT-PCR results demonstrated that in both spleen and head kidney tissues, mRNA expressions of typical Th1 immune response cytokines IL-2 and IFN-γ2 in ß-glucan group and anisodamine group were significantly higher than other groups and the level of immunoglobulins related to systemic immunity (IgM) and mucosal immunity (IgZ) were also enhanced in the immune period. DTT group slightly affected immune gene and serum enzyme activity, while did not show an adjuvant effect on survival rate. In addition, four adjuvant groups could obviously inhibit CyHV-2 replication. This study explored and proved the good efficiency of ß-glucan or anisodamine as immersion immune adjuvant and also provided reference for improving the efficiency of immersion immunity.


Assuntos
Doenças dos Peixes/prevenção & controle , Carpa Dourada , Infecções por Herpesviridae/veterinária , Herpesviridae/imunologia , Imunização/veterinária , Alcaloides de Solanáceas/imunologia , Vacinas Virais/imunologia , beta-Glucanas/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Aquicultura , Doenças dos Peixes/virologia , Carpa Dourada/imunologia , Carpa Dourada/virologia , Herpesviridae/fisiologia , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/virologia , Imunidade Inata , Imunidade nas Mucosas , Imunização/métodos , Propiolactona , Escopolamina/administração & dosagem , Escopolamina/imunologia , Alcaloides de Solanáceas/administração & dosagem , Taxa de Sobrevida , Vacinas de Produtos Inativados/administração & dosagem , Vacinas de Produtos Inativados/imunologia , Vacinas Virais/administração & dosagem , Replicação Viral , beta-Glucanas/administração & dosagem
3.
Artigo em Inglês | MEDLINE | ID: mdl-31707088

RESUMO

LEAP-2, a multifunctional peptide, not only exhibits a regulatory role in pathogenic infection, but also participates in the regulation of teleostean immunity. In this study, ORF sequence of WR-LEAP-2 was 240 bp and encoded 79 amino acid residues. Tissue-specific analysis revealed that the highest expression of WR-LEAP-2 was observed in liver. Aeromonas hydrophila challenge can sharply increase WR-LEAP-2 mRNA expression in liver, kidney and spleen. The purified WR-LEAP-2 peptide can directly bind to A. hydrophila and S. agalactiae, reduce the relative bacterial activity and limit bacterial growth in vitro. In addition, the treatment of WR-LEAP-2 can restrict bacterial dissemination in vivo and reduce production of pro-inflammatory cytokines. These results indicated that WR-LEAP-2 can confer protection against A. hydrophila- or S. agalactiae-stimulated MyD88-dependent pro-inflammatory cytokines activation.


Assuntos
Aeromonas hydrophila , Peptídeos Catiônicos Antimicrobianos/imunologia , Quimera/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/metabolismo , Carpa Dourada/imunologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Animais , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/farmacologia , Quimera/genética , Citocinas/metabolismo , Diploide , Doenças dos Peixes/genética , Carpa Dourada/genética , Infecções por Bactérias Gram-Negativas/genética , Fígado/imunologia
4.
Fish Shellfish Immunol ; 97: 72-82, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31846772

RESUMO

The aquaculture system based on biofloc technology (BFT) showed positive effects on prevention of Cyprinid herpesvirus 2 (CyHV-2) infection in gibel carp (Carassius auratus gibelio), which is detrimental to health and causes seriously economic losses to aquaculture. However, the enhancement mechanism of BFT regarding immunity and disease resistance of cultured species is scarce. Poly-ß-hydroxybutyrate (PHB) has been proved as one of bioactive compounds in bioflocs. In this study, two groups (4% PHB supplementation diets and control with basal diets) with 30-day feeding were set to study the effect of PHB supplementation on immune-related gene expression by qRT-PCR, time-course CyHV-2 replication in vivo by qPCR and intestinal microbiota by illumine high-throughput sequencing. PHB supplementation significantly up-regulated transcriptional levels of eight immune-related genes, decreased cumulative mortality of gibel carp and early CyHV-2 replication in spleen in vivo (P < 0.05). Additionally, PHB changed the microbial structure but not diversity, and significantly increased beneficial bacteria such as Bacillus sp. KEGG pathway analysis by PICRUSt demonstrated that oral administration of PHB up-regulated abundances of genes responsible for seven pathways and down-regulated genes in eleven pathways. Histological structures of foregut, mindgut and hindgut were also affected. Our findings suggested that profitable effects of PHB on immunity and disease resistance might be gut microbiota-related, and regulated through pathways of enzymes secretion, replication and repair, and host immune system. This study will provide new insights into understanding the enhancing mechanism of BFT on immunity and disease resistance of cultured animals, and developing prebiotics/probiotics-based immunotherapies to improve animal health and disease resistance.


Assuntos
Microbioma Gastrointestinal/efeitos dos fármacos , Carpa Dourada/imunologia , Infecções por Herpesviridae/veterinária , Herpesviridae/efeitos dos fármacos , Hidroxibutiratos/administração & dosagem , Imunidade Inata/genética , Poliésteres/administração & dosagem , Ração Animal/análise , Animais , Aquicultura , Suplementos Nutricionais , Resistência à Doença , Doenças dos Peixes/imunologia , Doenças dos Peixes/prevenção & controle , Doenças dos Peixes/virologia , Expressão Gênica , Carpa Dourada/genética , Herpesviridae/fisiologia , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/prevenção & controle , Replicação Viral/efeitos dos fármacos
5.
Fish Shellfish Immunol ; 94: 510-516, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31541778

RESUMO

Skin plays an important role in the innate immune responses of fish, particularly towards bacterial infection. To understand the molecular mechanism of mucosal immunity of fish during bacterial challenge, a de novo transcriptome assembly of crucian carp Carassius auratus skin upon Aeromonas hydrophila infection was performed, the latter with Illumina Hiseq 2000 platform. A total of 118111 unigenes were generated and of these, 9693 and 8580 genes were differentially expressed at 6 and 12 h post-infection, respectively. The validity of the transcriptome results of eleven representative genes was verified by quantitative real-time PCR (qRT-PCR) analysis. A comparison with the transcriptome profiling of zebrafish skin to A. hydrophila with regards to the mucosal immune responses revealed similarities in the complement system, chemokines, heat shock proteins and the acute-phase response. GO and KEGG enrichment pathway analyses displayed the significant immune responses included TLR, MAPK, JAK-STAT, phagosome and three infection-related pathways (ie., Salmonella, Vibrio cholerae and pathogenic Escherichia coli) in skin. To our knowledge, this study is the first to describe the transcriptome analysis of C. auratus skin during A. hydrophila infection. The outcome of this study contributed to the understanding of the mucosal defense mechanisms in cyprinid species.


Assuntos
Doenças dos Peixes/imunologia , Carpa Dourada/genética , Carpa Dourada/imunologia , Imunidade Inata/genética , Pele/metabolismo , Aeromonas hydrophila/fisiologia , Animais , Perfilação da Expressão Gênica/veterinária , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária
6.
Fish Shellfish Immunol ; 93: 879-887, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31421239

RESUMO

Cyprinid herpesvirus 2 (CyHV-2) is the causative pathogen of herpesviral haematopoietic necrosis disease, which has caused huge economic losses to aquaculture industry in China. In this study, nine truncated CyHV-2 membrane glycoproteins (ORF25, ORF25C, ORF25D, ORF30, ORF124, ORF131, ORF136, ORF142A, ORF146) and a GFP reporter protein were respectively expressed using baculovirus surface displaying system. Western blot showed that the proteins were successfully packaged in the recombinant virus particles. In baculovirus transduced gibel carp kidney cells, the target proteins were expressed and displayed on the fish cell surface. Healthy gibel carp were immunized by immersion with the recombinant baculoviruses and the fish treated with phosphate-buffered saline (PBS) were served as mock group. The expression of interleukin-11 (IL-11), interferon α (IFNα) and a complement component gene C3 were significantly up-regulated in most experimental groups, and interferon γ (IFNγ) expression in some groups were also induced after immunization. Subsequently, the immunized gibel carp were challenged by intraperitoneal injection of CyHV-2 virus. All the immunized groups exhibited reduced mortality after CyHV-2 challenge. In the groups immunized with baculoviruses displaying and expressing ORF25, ORF25C and ORF146, the relative percentage survival values reached 83.3%, 87.5% and 70.8%, respectively. Our data suggested that baculovirus-displayed ORF25, ORF25C and ORF146 could be potential vaccine candidates for the prevention of CyHV-2 infection in gibel carp.


Assuntos
Doenças dos Peixes/imunologia , Carpa Dourada/imunologia , Herpesviridae/fisiologia , Imunidade Inata , Imunização/veterinária , Proteínas da Matriz Viral/metabolismo , Animais , Baculoviridae/fisiologia , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/veterinária , Imersão , Imunização/métodos , Vírus Reordenados/fisiologia
7.
Fish Shellfish Immunol ; 92: 101-110, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31163296

RESUMO

Cyprinid herpesvirus II (CyHV-2) is highly contagious and pathogenic to Carassius auratus gibelio (gibel carp), causing enormous economic losses in aquaculture in Yancheng city, Jiangsu province, China; however, to date, there is no effective way to protect C. auratus gibelio from CyHV-2 infection. In this study, a recombinant baculovirus vector vaccine, BacCarassius-D4ORFs, containing a fused codon-optimized sequence D4ORFs comprising the ORF72 (region 1-186 nt), ORF66 (region 993-1197 nt), ORF81 (region 603-783 nt) and ORF82 (region 85-186 nt) genes of CyHV-2, driven by a Megalobrama amblycephala ß-actin promoter, was constructed. Then, qPCR, Western blotting and immunofluorescence assays showed that the fused gene D4ORFs was successfully delivered and expressed in fish cells or tissues by transduction with BacCarassius-D4ORFs. The fused gene D4ORFs could not be detected by PCR in the C. auratus gibelio injected with BacCarassius-D4ORFs after 7 weeks. Specific antibody against ORF72 could be detected in the serum of vaccinated C. auratus gibelio by injection with BacCarassius-D4ORFs. Furthermore, when C. auratus gibelio were vaccinated with BacCarassius-D4ORFs via the oral or injection route, followed by challenge with CyHV-2, the relative survival rate of immunized C. auratus gibelio reached 59.3% and 80.01%, respectively. These results suggested that BacCarassius-D4ORFs has the potential to be used as a vector-based vaccine for the prevention and treatment of disease caused by CyHV-2 infection.


Assuntos
Doenças dos Peixes/prevenção & controle , Carpa Dourada/imunologia , Herpesviridae/imunologia , Vacinas contra Herpesvirus/imunologia , Animais , Genes Virais , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/veterinária , Fases de Leitura Aberta , Vacinas Atenuadas/imunologia
8.
Fish Shellfish Immunol ; 91: 78-86, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31039439

RESUMO

Type I interferons, as a class of multipotent cytokines, play a key role in host antiviral immune responses. In this study, a type I IFN coding gene of gibel carp, Carassius auratus gibelio, CagIFNa was cloned and sequenced. The full-length cDNA sequence of CagIFNa consists of 724 nucleotides that encode a predicted protein of 183 amino acids. CagIFNa has two highly conserved cysteine residues in the deduced protein, which is mostly conserved in the fish group I type I IFNs. CagIFNa was identified as a member of the IFNa subgroup of group I type I IFNs by phylogenetic analysis. CagIFNa transcripts were detected in all investigated tissues with higher levels in the liver, intestine, spleen and head kidney of gibel carp. Following injection with Cyprinid herpesvirus 2 (CyHV-2), CagIFNa gene expression was significantly inhibited in the spleen but delayed and then increased in head kidneys. Similarly, while CagIFNa expression was rapidly induced in gibel carp brain (GiCB) cells by poly I:C stimulation and its high induction level was delayed following CyHV-2 infection. CagIFNa overexpression in GiCB cells drastically reduced virus CPE and titer. Furthermore, several genes associated with type I IFN signaling pathway including IRF3, IRF7, IRF9, STAT1, Mx1 and PKR were induced in GiCB cells overexpressing CagIFNa upon CyHV-2 infection. These results show that CagIFNa plays a role in antiviral immune system in gibel carp.


Assuntos
Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Carpa Dourada/genética , Carpa Dourada/imunologia , Imunidade Inata/genética , Interferon Tipo I/genética , Interferon Tipo I/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Herpesviridae , Infecções por Herpesviridae/imunologia , Interferon Tipo I/química , Filogenia , Poli I-C/farmacologia , Distribuição Aleatória , Alinhamento de Sequência/veterinária
9.
Fish Shellfish Immunol ; 89: 141-148, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30926477

RESUMO

Cadmium (Cd) is the most common heavy metal and is easily detected in aquatic environments worldwide. The genus Bacillus was one of dominant probiotics, which was commonly used in aquaculture. The present study was undertaken to explore the effects of Bacillus cereus (B. cereus) supplementation on hematological parameters and the immune response of Carassius auratus gibelio (C. gibelio) following Cd exposure. Fish were exposed to waterborne Cd (0, 1 and 2 mg/L) and/or treated with dietary B. cereus at 108 cfu/g for four weeks. The hematological disturbances observed after exposure of waterborne Cd included significant decreases in red blood cell (RBC) count, hemoglobin (Hb) concentration and hematocrit (HCT). While significant elevation (P < 0.05) of RBC count, HCT and Hb levels in the 1 and 2 mg/L Cd-B. cereus administration group at 4 weeks, compared with the Cd-only group. Among serum enzymatic activities, aspartate aminotransferase (AST) and alanine transaminase (ALT) activities by Cd exposure were significantly higher than controls, but this increase was effectively inhibited in Cd-B. cereus administration groups. In the Cd-B. cereus administration group, significant down-regulation of Hsp70, Hsp90, IL-1ß, IL-6, IL-10 and TNF-α in conjunction with the up-regulation of IgM and LZM in the spleen indicated that B. cereus alleviated the Cd-induced damage to the immune system to some degree. The results of this study suggested that B. cereus has the potential to countermeasure Cd-induced hematological disturbances and immunosuppression in C. gibelio.


Assuntos
Bacillus cereus/química , Cádmio/efeitos adversos , Carpa Dourada/imunologia , Tolerância Imunológica , Probióticos/farmacologia , Poluentes Químicos da Água/efeitos adversos , Ração Animal/análise , Animais , Dieta/veterinária , Testes Hematológicos/veterinária , Distribuição Aleatória
10.
J Immunol ; 202(8): 2407-2420, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30850476

RESUMO

In mammals, tripartite motif (TRIM) proteins have emerged as pivotal players endowed with, directly, antiviral effects and, indirectly, modulatory capacity of the innate immune response. An unprecedented expansion of TRIM family has occurred in fish; however, the functional role of fish TRIM family members remains largely unknown. In this study, we identify a species-specific TRIM gene from crucian carp Carassius auratus, named FTRCA1, phylogenetically similar to the members of finTRIM, a subfamily of TRIM exclusively in teleost fish. FTRCA1 is induced by IFN and IFN stimuli as a typical IFN-stimulated gene. Overexpression of FTRCA1 negatively regulates IFN antiviral response by inhibition of IRF3 phosphorylation; consistently, knockdown of FTRCA1 results in enhanced levels of IRF3 phosphorylation and also IFN expression following poly(I:C) transfection. Whereas FTRCA1 is associated with several pivotal signaling molecules of RIG-I-like receptor pathway, its association with TBK1 results in autophage-lysosomal degradation of TBK1, thus abrogating the downstream IFN induction. Interestingly, FTRCA1 is phosphorylated by TBK1, but this phosphorylation is not required for downregulation of TBK1 protein. Transfection assays indicate that FTRCA1 is likely an E3 ligase with the requirement of RING finger domain, and deletion of N-terminal RING domain or mutation of seven conservative sites abolishes the negative regulatory function of FTRCA1. Collectively, these results illuminate a novel finTRIM-mediated innate immune modulatory pathway, thus providing insights into species-specific regulation of fish IFN response.


Assuntos
Autofagossomos/imunologia , Proteínas de Peixes/imunologia , Carpa Dourada/imunologia , Interferons/imunologia , Lisossomos/imunologia , Proteínas Serina-Treonina Quinases/imunologia , Proteólise , Proteínas com Motivo Tripartido/imunologia , Animais , Proteínas de Peixes/genética , Carpa Dourada/genética , Células HEK293 , Humanos , Interferons/genética , Lisossomos/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas com Motivo Tripartido/genética
11.
Dev Comp Immunol ; 94: 11-15, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30659854

RESUMO

Hepcidin is an antimicrobial peptide and an iron regulatory protein that prevents the release of excess iron in the blood. There is evidence suggesting that teleost hepcidin is a major player in antimicrobial defense against various bacteria species, but little is known regarding the effects of teleost hepcidin in protozoan parasitic infections. We examined the role of hepcidin during the course of infection of goldfish with Trypanosoma carassii. Quantitative real-time PCR was used to determine the expression of hepcidin in goldfish immune organs during the course of T. carassii infection. During the acute phase of the T. carassii infection, the mRNA levels of hepcidin were up-regulated in liver and kidney. In contrast, an up-regulation of hepcidin mRNA expression in spleen was observed during the chronic phase of the infection. Furthermore, a synthetic goldfish hepcidin peptide induced trypanosome lysis in vitro, and parasite surface disruption was confirmed by scanning electron microscopy (SEM) analysis. These results suggest that, in addition to well-characterized direct antibacterial activities, teleost hepcidin also exhibits trypanocidal activity.


Assuntos
Anti-Infecciosos/metabolismo , Doenças dos Peixes/imunologia , Carpa Dourada/imunologia , Hepcidinas/metabolismo , Macrófagos/imunologia , Trypanosoma/imunologia , Tripanossomíase/imunologia , Animais , Citocinas/metabolismo , Imunidade Inata , Transcriptoma , Regulação para Cima
12.
Fish Shellfish Immunol ; 86: 913-921, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30550991

RESUMO

The present study was conducted to evaluate dietary inosine 5'-monophosphate (5'-IMP) on growth, immune genes expression and disease resistance against Aeromonas hydrophila in juvenile gibel carp (Carassius auratus gibelio var. CAS Ⅲ) (initial body weight: 7.48 g). Six diets were formulated containing exogenous 5'-IMP at three gradient levels (0, 0.1% and 0.2%) in the high dietary fishmeal group (15% fishmeal: D1, D2, D3) and in the high dietary soybean meal group (33% soybean meal: D4, D5, D6). Each diet was randomly allotted to triplicate tanks in a recirculating system. After the feeding trial, fish were exposed to Aeromonas hydrophila challenge. Hematological and immunological responses were analyzed before and after challenge. The results indicated that feeding rate in all 5'-IMP supplemented treatments (D2, D3, D5 and D6) and daily growth coefficient in D5 and D6 were reduced compared with those of respective control treatments (D1 and D4) without 5'-IMP addition (P < 0.05). The cumulative survival rates were numerically improved by dietary 5'-IMP supplementation (P > 0.05). Compared with the respective control treatment, in the high fishmeal group, plasma SOD and MPO were significantly elevated in D3 at the end of feeding trial (P < 0.05), plasma SOD and lysozyme were significantly increased in D3 after bacterial challenge (P < 0.05); in high soybean meal group, plasma lysozyme activity was significantly elevated in D5 post bacterial challenge (P < 0.05). Most of the expression of immune related genes (intelectin, major histocompatibility complex class II ß (MHC II ß), Complement 3 (C3), Complement component C7-1 (ccC7), lysozyme C, Interleukin 1ß (IL-1ß), Tumor necrosis factor α1 (TNF-α1), Transforming growth factor-beta (TGF-ß) and Interleukin 8 (IL-8)) in spleen, kidney and liver of the fish were significantly affected by supplementation of 5'-IMP at the end of feeding trial and post bacterial challenge. Additionally, adding 5'-IMP in high soybean meal diets exerted further effects of promoting immunity than counterparts in high fishmeal diets. Considering enhanced disease resistance, the immunopotentiation of 5'-IMP was manifested when the addition level was 0.1% in high soybean meal diets and 0.2% in high fishmeal diets.


Assuntos
Dieta/veterinária , Regulação da Expressão Gênica/imunologia , Carpa Dourada/genética , Carpa Dourada/imunologia , Inosina Monofosfato/metabolismo , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Dieta/classificação , Suplementos Nutricionais/análise , Resistência à Doença/efeitos dos fármacos , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Carpa Dourada/crescimento & desenvolvimento , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Inosina Monofosfato/administração & dosagem
13.
Fish Physiol Biochem ; 45(1): 427-437, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30361821

RESUMO

ß-Thymosins play critical roles in the regulation of many important physiological processes, but their function in teleost fishes remains poorly understood. In this study, the full-length cDNA coding for a thymosin ß (Tß) was cloned and identified in goldfish, Carassius auratus (gfTß). The gfTß cDNA consisted of 653 bp with an open reading frame of 135 bp that encodes a 44 amino acid polypeptide. Sequence analysis revealed one thymosin domain and a highly conserved actin-binding motif (18LKKTET23). Expression of gfTß transcript was detected ubiquitously in all tissues examined, with relatively higher levels in the brain, intestine, spleen, gill, skin, kidney, and testis. Cadmium and H2O2 exposure induced increases in gfTß transcript levels in the liver and spleen. Moreover, gfTß transcription was upregulated in response to LPS challenge in the spleen while Poly I:C treatment did not affect gfTß expression. In vivo injection of recombinant gfTß generated from an Escherichia coli system induced expression of T lymphocyte-related genes (RAG1 and CD8α). These results suggest that gfTß may be involved in the immune response of teleost fishes via modulation of T lymphocyte development.


Assuntos
Clonagem Molecular , Regulação da Expressão Gênica/fisiologia , Carpa Dourada/metabolismo , Timosina/metabolismo , Animais , Carpa Dourada/genética , Carpa Dourada/imunologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Timosina/genética
14.
Dev Comp Immunol ; 91: 93-100, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30385316

RESUMO

In this study, we investigated the immune responses against Mycobacterium gordonae in ginbuna crucian carp. Cumulative mortality of ginbuna injected with 2.0 × 107 CFU of M. gordonae was 50% at 170 days post-infection. CD4-1, CD8α, T-bet and IFNγ2 gene expression levels were significantly upregulated in ginbuna injected with 1.9 × 108 CFU of M. gordonae at 21 and 28 days post-infection. The CD4-2 level did not change during the experiment. Granulomatous responses consisted of central macrophage accumulation and surrounding lymphocytes, and Ziehl-Neelsen-positive bacteria were observed in the trunk kidney of the challenged fish. Immunohistochemistry using anti-ginbuna IFNγs and anti-ginbuna CD4-1 polyclonal antibody revealed that the marginal lymphocytes were positive for CD4-1, and the IFNγ-producing cells surrounded the mycobacterial cell-laden phagocytes. These results suggest that CD4-1+ cells and IFNγ2 play important roles in the granulomatous inflammation against Mycobacterial infections in teleosts.


Assuntos
Doenças dos Peixes/imunologia , Carpa Dourada/imunologia , Granuloma/imunologia , Inflamação/imunologia , Linfócitos/imunologia , Macrófagos/imunologia , Infecções por Mycobacterium não Tuberculosas/imunologia , Micobactérias não Tuberculosas/fisiologia , Animais , Antígenos CD4/metabolismo , Antígenos CD8/metabolismo , Proteínas de Peixes/metabolismo , Imuno-Histoquímica , Interferon gama/metabolismo , Proteínas com Domínio T/metabolismo
15.
Fish Shellfish Immunol ; 84: 458-469, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30312666

RESUMO

Probiotic strains play an increasing role in the production of healthy animals used as a food source. Elucidating the mechanisms of action that allow probiotic-driven immunomodulation may facilitate different applications such as the prevention of infectious diseases in food organisms. This study elucidates the probiotic effects of Exiguobacterium acetylicum S01 on the growth, haematological profile, innate immune capacity, expression of cytokine genes, and resistance to diseases of Carassius auratus caused by Aeromonas hydrophila infection. Three fish groups were fed with the following diets containing different doses of E. acetylicum S01 (CFU g-1): basal diet 0 (BD, without probiotic), 2.5 × 107 (DI) and 2.7 × 109 (DII)-CFU g-1 for 4 weeks. After 4 weeks, the fish were injected intraperitoneally with A. hydrophila and the percentage of survival was recorded over 21 days of post-challenge. Results revealed that dietary supplementation of E. acetylicum S01 significantly (P < 0.05) enhanced the growth, haematological profile and cellular immune responses including respiratory burst, phagocytic activities and antimicrobial enzymes (myeloperoxidase and lysozyme) and total immunoglobulin levels were improved by probiotic feeding at both occasions. Comparatively, expression of c- and g-type lysozyme followed by pro- and anti-inflammatory cytokines (IL-1ß, IL-10 and TGFß) was up-regulated in kidney, head-kidney and spleen. Moreover, fish fed with diet DII had a significantly higher (P < 0.05) survival rate (73.2%) after challenging. The survival rate was only 33.2% of the BD group against A. hydrophila infection. Our results revealed that E. acetylicum S01 delivered probiotic in feed exerts its influence on growth performance and provides disease resistance by stimulating the immune system at the cellular and molecular levels in C. auratus.


Assuntos
Bacillales/química , Resistência à Doença/efeitos dos fármacos , Doenças dos Peixes/prevenção & controle , Regulação da Expressão Gênica/efeitos dos fármacos , Carpa Dourada/imunologia , Imunidade Inata/efeitos dos fármacos , Probióticos/farmacologia , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Citocinas/genética , Citocinas/metabolismo , Dieta/veterinária , Relação Dose-Resposta a Droga , Doenças dos Peixes/microbiologia , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Carpa Dourada/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Infecções por Bactérias Gram-Negativas/veterinária
16.
Fish Shellfish Immunol ; 82: 400-407, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30144566

RESUMO

A 50-day feeding trial was carried out to evaluate the partial replacement of fishmeal by yeast culture (YC) on growth performance, immune response and resistance against Aeromonas hydrophila in gibel carp CAS Ⅲ (Carassius auratus gibelio). Four isonitrogenous and isoenergetic practical diets including a basal diet (the control diet containing 10% fish meal, D0) and three yeast culture diets (substituting 20%, 40%, 60% of the fishmeal in the basal diet, D20, D40 and D60, respectively) were formulated. Each diet was randomly allocated to quadruplicate fish groups (average initial body weight: 28.70 ±â€¯0.03 g) reared in a recirculating system. After the growth trial, bacterial challenge test was conducted. The results showed that no noteworthy variations in feed intake, growth performance and morphology indices were found among groups (P > 0.05). YC Supplemented diet exerted little significant influence on plasma parameters including triglyceride, glucose, creatinine, total protein and urea nitrogen compared with the control group (P > 0.05). No obvious variations were found in activities of plasma lysozyme, IgM, MPO and SOD before challenge test among dietary treatments (P > 0.05), whereas considerable higher value of the foresaid indicators was discovered in D40 after bacteria challenge (P < 0.05). Transcriptional levels of Toll like receptor 2 (TLR2), myeloid differentiation factor 88 (MyD88), Toll/IL-1 receptor domain-containing adaptor protein (TIRAP) and interleukin-1ß (IL-1ß) in spleen after challenge were significantly up-regulated in D40 compared with D0 (P < 0.05). Cumulative survival rate in D40 and D60 were significantly higher than those in D0 and D20 (P < 0.05). Taken together, yeast culture could be a suitable fishmeal alternative in diets of gibel carp and dietary inclusion of 4 g YC per 100 g diet enhanced the immunity and disease resistance of gibel carp partly via TLR2 pathway.


Assuntos
Resistência à Doença/imunologia , Doenças dos Peixes/imunologia , Carpa Dourada/imunologia , Fermento Seco/farmacologia , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Dieta/veterinária , Carpa Dourada/crescimento & desenvolvimento , Infecções por Bactérias Gram-Negativas/imunologia , Distribuição Aleatória
17.
Ecotoxicol Environ Saf ; 161: 755-762, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29957583

RESUMO

Lead (Pb) poisoning in humans and fish represents a significant global problem. Bacillus subtilis (B. subtilis) is a widely used probiotic in aquaculture. Carassius auratus gibelio (C. gibelio) is one of the most important aquaculture species with great commercial value. The objective of this study was to evaluate the potential of B. subtilis in ameliorating lead-induced toxicity in C. gibelio. The fish were exposed for 60 days to waterborne Pb at 0, 0.05, 0.5 and 1 mg/L and/or dietary B. subtilis at 109 cfu/g. After 30 and 60 days, the fish were sampled and bioaccumulation, antioxidant activity and immune responses were assessed. The results revealed that B. subtilis confers significant protective effects against lead toxicity by preventing alterations in the levels of bioaccumulation, superoxide dismutase, catalase and glutathione. B. subtilis also assists in the recovery of blood δ-aminolevulinic acid dehydratase, lysozyme, and IgM levels while regulating the expression of immune-related genes including IL-10, lysozyme, TNF-α, IgM and Hsp70 after 60 days of lead exposure. Our results suggest that administration of B. subtilis (109 cfu/g) has the potential to combat lead toxicity in C. gibelio.


Assuntos
Bacillus subtilis , Carpa Dourada/metabolismo , Chumbo/toxicidade , Probióticos , Animais , Antioxidantes/metabolismo , Catalase/metabolismo , Glutationa/metabolismo , Carpa Dourada/sangue , Carpa Dourada/imunologia , Imunoglobulina M/sangue , Muramidase/sangue , Sintase do Porfobilinogênio/sangue , Superóxido Dismutase/metabolismo
18.
Fish Shellfish Immunol ; 79: 265-273, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29775741

RESUMO

The present study examined the effect of dietary spirulina, Arthrospira platensis on growth performance, blood physiological indices, immune-related gene expressions and resistance of juvenile gibel carp against Aeromonas hydrophila infection. Four isonitrogenous (360 g kg-1) and isolipidic (90 g kg-1) diets were formulated with containing different levels of spirulina powder of 0 g (SP0, the control diet), 3.38 g (SP3.38), 6.76 g (SP6.76) and 13.52 g (SP13.52) per 100 g diet to replace 0%, 25%, 50% and 100% of fishmeal protein, respectively. And each diet was randomly assigned to triplicate tanks (150-L capacity per each) and each tank was stocked with 22 fish (15.37 ±â€¯0.06 g). Fish were fed one of the tested diets up to satiation twice a day for 46 days. A challenge test was carried out after the feeding trial by injecting Aeromonas hydrophila intraperitoneally for 7 days. The results showed that fish growth, feeding rate in groups SP3.38 and SP6.76 were significantly higher than those of groups SP0 and SP13.52 (P < 0.05). Feed efficiency and protein retention rate had no significant difference among all tested groups. Plasma superoxide dismutase and phagocyte activity of blood leukocytes significantly increased in the spirulina-fed fish groups at 12-h post the bacterial challenge (P < 0.05). Both pre and post challenge test, plasma lysozyme activities in spirulina-fed groups were significantly higher than that in the control group (P < 0.05). Plasma malondialdehyde got the lowest value in the SP13.52 group before and after the challenge test. The transcriptional levels of TLR2 (Toll like receptor 2), myeloid differentiation factor 88 (MyD88), Toll/IL-1 receptor domain-containing adaptor protein (TIRAP), interleukin-1ß (IL-1ß) and tumor necrosis factor-α1 (TNF-α1) in spleen and kidney significantly increased post the bacterial challenge compared to the pre challenge. And the relative expressions of the immune-related genes of spirulina-fed fish groups were higher than those of the control group before and after the challenge test. The 7-day cumulative survival rate after the bacterial challenge was highest in the SP3.38 group (P < 0.05). The present results indicated that low dietary inclusion of spirulina significantly enhanced the immune response of gibel carp partly through TLR2 pathway and 3.38% of dietary spirulina was recommended for the juveniles based on the growth and immune response.


Assuntos
Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Regulação da Expressão Gênica/imunologia , Carpa Dourada/genética , Carpa Dourada/imunologia , Spirulina/química , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Resistência à Doença/imunologia , Proteínas de Peixes/metabolismo , Carpa Dourada/crescimento & desenvolvimento , Infecções por Bactérias Gram-Negativas/imunologia
19.
J Parasitol ; 104(4): 353-358, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29648929

RESUMO

Species composition and diversity of dactylogyrids were compared on gills of wild and cultured goldfish (silver crucian carp) Carassius auratus from 3 naturally populated lakes and 3 stocked aquaculture ponds in the Hubei province of China to examine the differences in the gill parasite community between these natural and farmed waters. Of the 7 Dactylogyrus spp. detected, all were found in lakes and 5 in ponds, with Dactylogyrus inexpectatus and Dactylogyrus anchoratus being absent from ponds. No significant correlation was found between the species richness and habitat area or host size, nor was there a significant difference in mean species richness between lakes (0.41-0.65) and ponds (0.30-0.76). Brillouin's diversity in lakes (0.049-0.067) was higher than that in ponds (0.024-0.046), but not significantly so. Although the diversity of parasite communities was higher in wild goldfish, higher mean abundance of some Dactylogyrus spp. was found in cultured goldfish. Based on Bray-Curtis similarity, it was difficult to differentiate parasite communities in lakes from those in ponds at the infracommunity level, whereas the 3 lakes and Guanqiao pond differed markedly from the remaining 2 ponds at the component community level. Although infracommunities differed among waterbodies, no effects of fish length or waterbody type were found on infracommunity or component community structure. Together, these results suggest that abundance and species richness of Dactylogyrus spp. on goldfish in lakes and farm ponds are influenced by habitat-specific environmental factors.


Assuntos
Doenças dos Peixes/parasitologia , Carpa Dourada/parasitologia , Platelmintos/classificação , Infecções por Trematódeos/veterinária , Análise de Variância , Animais , Animais Selvagens , China/epidemiologia , Doenças dos Peixes/epidemiologia , Doenças dos Peixes/imunologia , Pesqueiros , Brânquias/parasitologia , Carpa Dourada/imunologia , Lagos , Platelmintos/crescimento & desenvolvimento , Tanques , Densidade Demográfica , Prevalência , Infecções por Trematódeos/epidemiologia , Infecções por Trematódeos/imunologia , Infecções por Trematódeos/parasitologia
20.
Fish Shellfish Immunol ; 78: 322-330, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29702236

RESUMO

This study evaluated the probiotic potential of B. velezensis JW through experimental and genomic analysis approaches. Strain JW showed antimicrobial activity against a broad range of fish pathogenic bacteria including Aeromonas hydrophila, Aeromonas salmonicida, Lactococcus garvieae, Streptococcus agalactiae, and Vibrio Parahemolyticus. Fish (Carassius auratus) were fed with the diets containing 0 (control), 107, and 109 cfu/g of B. velezensis JW for 4 weeks. Various immune parameters were examined at 1, 2, 3, and 4 weeks of post-feeding. Results showed that JW supplemented diets significantly increased acid phosphatase (ACP), alkaline phosphatase (AKP), and glutathione peroxidase (GSH-PX) activity. The mRNA expression of immune-related genes in the head kidney of C. auratus was measured. Among them, the interferon gamma gene (IFN- γ) and tumor necrosis factor-α (TNF-α) showed higher expression after 3 and 4 weeks of feeding (P < 0.05). The expression of interleukin-1 (IL-1) only being significantly upregulated by 109 cfu/g of JW after 1 week of feeding (P < 0.05). The upregulation of interleukin-4 (IL-4) increased over time from 1st to 4th week. The expression of interleukin-10 (IL-10) and interleukin-12 (IL-12) showed an opposite expression pattern with IL-10 significantly upregulated and IL-12 significantly downregulated by JW containing diets at 2, 3, and 4 weeks of post-feeding (P < 0.05). Moreover, fish fed with JW supplemented diets showed significantly improved survival rate after A. hydrophila infection. The analysis of the genome of JW revealed several features aiding host health and being relevant to the GIT adaptation. Four bacteriocins, three Polyketide Synthetase (PKS), and five Nonribosomal Peptide-Synthetase (NRPS) gene clusters were identified in the genome. In summary, the above results clearly proved that B. velezensis JW has the potential to be developed as a probiotic agent in aquaculture.


Assuntos
Bacillus/química , Doenças dos Peixes/imunologia , Proteínas de Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Carpa Dourada/imunologia , Imunidade Inata/genética , Probióticos/farmacologia , Ração Animal/análise , Animais , Infecções Bacterianas/imunologia , Fenômenos Fisiológicos Bacterianos , Dieta/veterinária , Proteínas de Peixes/genética , Carpa Dourada/genética
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