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1.
Mol Immunol ; 119: 8-17, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31927202

RESUMO

Motile sperm domain containing 2 (MOSPD2) is a single-pass membrane protein to which until recently little function had been ascribed. Although its mammalian homologs have been identified, the status of the mospd2 gene in lower vertebrates is still unknown. In the present study, cDNA of the mospd2 gene of barbel steed (Hemibarbus labeo) was cloned and sequenced to characterize its potential involvement in the innate immune system of this fish. Sequence analysis revealed that the predicted barbel steed MOSPD2 protein contained an N-terminal extracellular portion composed of a CRAL-TRIO domain, a motile sperm domain, and a transmembrane domain, as well as a short C-terminal intracellular domain. Phylogenetic tree analysis indicated that barbel steed MOSPD2 is closely related to that of zebrafish. Barbel steed mospd2 transcripts were detected in a wide range of tissues, with the highest level being found in the gill. In response to lipopolysaccharide (LPS) treatment or Aeromonas hydrophila infection, mospd2 gene expression was significantly altered in the head kidney, spleen, and mid-intestine. The expression of mospd2 gene was detected in monocytes/macrophages (MO/MФ), neutrophils, and lymphocytes, and was found to be mainly expressed in MO/MФ. At the same time, using flow cytometry, we also confirmed that MOSPD2 protein is located on MO/MФ, neutrophil, and lymphocyte membranes. Following treatment with LPS or A. hydrophila, MOSPD2 protein expression was induced in these immune cells. The migration of MO/MФ and neutrophils decreased significantly upon MOSPD2 blockade with anti-MOSPD2 IgG in a dose-dependent manner, whereas this treatment had no significant effect on lymphocytes migration. To the best of our knowledge, our study, for the first time, provides evidence that MOSPD2 mediates the migration of MO/MФ and neutrophils in a fish species.


Assuntos
Quimiotaxia/fisiologia , Cyprinidae/fisiologia , Proteínas de Peixes/fisiologia , Proteínas de Membrana/fisiologia , Aeromonas hydrophila/imunologia , Animais , Clonagem Molecular , Cyprinidae/genética , Cyprinidae/imunologia , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Macrófagos/fisiologia , Proteínas de Membrana/genética , Monócitos/fisiologia , Neutrófilos/fisiologia , Análise de Sequência de DNA
2.
Mol Immunol ; 119: 18-26, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31954283

RESUMO

Fish interferon (IFN)-mediated antiviral innate immunity is the first line of defense against virus invasion. In the present study, we identify two fish IFN genes (here tentatively named IFNa and IFNc) with different-sized 3' UTRs from clone F strain of gibel carp Carassius auratus gibelio. Carp IFNa has a relatively short 3'UTR without AU-rich elements (AREs) but IFNc has a long one with 9 AREs. Functionally, carp IFNa and IFNc display significantly antiviral potential to viral infection, likely through induction of downstream IFN-stimulated genes (ISGs). Both carp IFN genes are induced by viral infection, poly(I:C) treatment and IRF3/7, which are ascribed to the IFN-sensitive response elements (ISRE) within their promoters. Carp IFN genes are also induced by each other and by themselves, indicating existence of a positive feedback loop in fish IFN-mediated antiviral immune response. Comparative analyses of 3'UTR-mediated expression regulation at mRNA and protein levels show that the ARE-containing 3'UTR of carp IFNc rather than the short 3'UTR of carp IFNa promotes mRNA decay but instead results in high-level protein expression, indicating that 3'UTR of fish IFN mRNAs might be a potential factor for regulation of IFN-mediated antiviral immune response. Considering a fact that a given protein function is largely related to its protein level, these results suggest that both promoter and 3'UTR contribute to the transcription and translation of fish IFN genes, thus shaping their eventually antiviral potential.


Assuntos
Regiões 3' não Traduzidas , Doenças dos Peixes/genética , Carpa Dourada/genética , Interferons/genética , Regulação para Cima , Elementos Ricos em Adenilato e Uridilato , Animais , Linhagem Celular , Doenças dos Peixes/imunologia , Infecções por Herpesviridae/genética , Infecções por Herpesviridae/veterinária , Infecções por Herpesviridae/virologia , Especificidade da Espécie
3.
J Fish Dis ; 42(12): 1657-1666, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31591734

RESUMO

Streptococcus iniae is one of the most serious aquatic pathogens, causing significant economic losses in marine and freshwater species, including Asian seabass (Lates calcarifer). Controlling this gram-positive bacterial pathogen has been an issue in aquaculture systems, due to the combined effects of aquaculture intensification and climatic impacts. To date, there have not been any genetic parameter estimates for S. iniae resistance in Asian seabass. The main aim of this study was to examine genetic variation in S. iniae resistance and its genetic correlations with growth and cannibalism in Asian seabass families produced from a breeding programme for high growth in 2016 and 2017. The study included a total of 5,835 individual fish that were offspring of 41 sires and 60 dams (31 half-sib and 34 full-sib families). The experimental fish were challenged by intraperitoneal injection with a volume containing 105  CFU (colony-forming unit)/fish. Resistance to S. iniae was measured as survival rate at 6 hr, 3, 5, 7, 10 and 15 days post-challenge test. There were significant variations in S. iniae resistance among families at different observation periods (ranging from 24.4% to 80%). Restricted maximum-likelihood method and mixed model analysis were applied to estimate heritability for S. iniae resistance. The heritability for S. iniae resistance ranged from 7% to 18% across different statistical models used. The common full-sib effects accounted for 0.1%-2% of the total variation in resistance to S. iniae. Genetic correlations of the S. iniae resistance at 6 hr and 3 days with later post-challenge test periods were low to moderate. However, these estimates for S. iniae resistance between successive measurement times (5, 7, 10 and 15 days) were high and close to 1. The genetic correlations of resistance with body weights at 180, 270 and 360 days post-hatch were not significant as well with cannibalism. It is concluded that there is substantial additive genetic variation in resistance to S. iniae, suggesting there is potential for genetic improvement of Asian seabass for resistance to S. iniae through selective breeding.


Assuntos
Resistência à Doença/genética , Doenças dos Peixes/genética , Perciformes/genética , Infecções Estreptocócicas/veterinária , Animais , Aquicultura , Canibalismo , Feminino , Doenças dos Peixes/microbiologia , Masculino , Modelos Estatísticos , Perciformes/microbiologia , Fenótipo , Infecções Estreptocócicas/genética , Infecções Estreptocócicas/microbiologia , Streptococcus iniae
4.
Aquat Toxicol ; 215: 105282, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31509759

RESUMO

Aquatic ecosystems are now chronically polluted by a cocktail of many chemical substances. There is now clear evidence of associations between exposure to pollutants and greater susceptibility to pathogens. The aim of the present study was to characterize the defense capacities of rainbow trout (Oncorhynchus mykiss), chronically exposed to pendimethalin (PD), to subsequent experimental challenge with the infectious hematopoietic necrosis virus (IHNV). Immunological responses were examined at different organizational levels, from individuals to gene expression. No negative effects of PD were noted on the Fulton index nor on the liver or spleen somatic indices (LSI; SSI) before viral infection, but the infectious stress seems to generate a weak but significant decrease in Fulton and LSI values, which could be associated with consumption of energy reserves. During the viral challenges, the distribution of cumulative mortality was slightly different between infected groups. The impact of the virus on fish previously contaminated by PD started earlier and lasted longer than controls. The proportion of seropositive fish was lower in the fish group exposed to PD than in the control group, with similar quantities of anti-IHNV antibodies secreted in positive fish, regardless of the treatment. While no significant differences in C3-1 expression levels were detected throughout the experiment, TNF1&2, TLR3, Il-1ß and IFN expression levels were increased in all infected fish, but the difference was more significant in fish groups previously exposed to herbicide. On the other hand, ß-def expression was decreased in the pendimethalin-IHNV group compared to that in fish only infected by the virus (control-IHNV group).


Assuntos
Herbicidas/toxicidade , Imunidade Celular/efeitos dos fármacos , Imunidade Humoral/efeitos dos fármacos , Vírus da Necrose Hematopoética Infecciosa/fisiologia , Oncorhynchus mykiss/genética , Oncorhynchus mykiss/imunologia , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/veterinária , Compostos de Anilina/toxicidade , Animais , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Regulação da Expressão Gênica/efeitos dos fármacos , Oncorhynchus mykiss/virologia , Infecções por Rhabdoviridae/genética , Infecções por Rhabdoviridae/patologia , Poluentes Químicos da Água/toxicidade
5.
Fish Shellfish Immunol ; 94: 294-307, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31491530

RESUMO

Aeromonas salmonicida is a ubiquitous fish pathogen known to cause furunculosis. With the emergence of new subtypes and the expansion of the host range, it has threatened the health of a variety of marine and freshwater fish, particularly the non-salmonids, manifesting differently from the classical furunculosis. Although there have been reports of infection by atypical strains on the crucian carp, the pathogenesis and tissue pathology remain unclear. In this study, transcriptomics and histopathology were used to analyze the immune response and lesions of crucian carp infected with A. salmonicida. Comparative analysis showed 6579 differentially expressed genes (DEGs) (3428 down-regulated and 3151 up-regulated) were identified on day 5 post-infection (5 dpi). Further annotation and analysis revealed that the DEGs were enriched in enzyme regulator activity, response to oxidative stress, iron ion homeostasis and other functions, and mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB), toll-like receptor (TLR), and nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) etc., and immune-related signaling pathways. Meanwhile, the four C-type lysozyme genes found in all DEGs were significantly up-regulated after infection. In addition, there was severe bleeding on the body of the infected fish. Also, the intestine, liver, spleen, and kidney showed varying degrees of inflammatory damage, especially the goblet cell hyperplasia of intestinal mucosa epithelium and degeneration and necrosis of renal tubular epithelium cells. Additionally, with the increase in pathogen concentration, the cumulative mortality increased, the severity of lesions in the hindgut and head-kidney tissues increased. The relative expression levels of four immune-related genes (TNF-α, IL-1ß, IL-11, C-lysozyme) were also significantly upregulated, compared with the control (P < 0.05). In conclusion, this study provides a scientific basis for further study on the immune response, pathological diagnosis, and prevention of crucian carp infection caused by atypical A. salmonicida.


Assuntos
Aeromonas salmonicida/fisiologia , Carpas , Doenças dos Peixes/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Imunidade Inata/genética , Transcriptoma/imunologia , Imunidade Adaptativa/genética , Aeromonas salmonicida/classificação , Animais , Doenças dos Peixes/genética , Infecções por Bactérias Gram-Negativas/imunologia , Filogenia
6.
Fish Shellfish Immunol ; 94: 258-263, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31513913

RESUMO

Grass carp septicemia is a systemic inflammatory response that develops following a bacterial infection. The hyperinflammatory state develops could lead to septic shock and lethality. There is increasing evidence that microRNAs are involved in the regulation of the inflammatory response. In the present study, miR-21 was confirmed to be involved in the inflammatory response following infection with Aeromonas hydrophila and LPS stimulation. Both jnk and ccr7 were identified as target gene of miR-21 by overexpression, inhibition, and dual luciferase reporter assays experiments. Meanwhile, miR-21 targets the jnk and ccr7 to modulate downstream pro-inflammatory factors tnf-α, il-1ß, il-6, and il-12. Our results provide a theoretical basis for exploring the molecular mechanism of grass carp miR-21 regulating inflammation.


Assuntos
Carpas/genética , Carpas/imunologia , Doenças dos Peixes/microbiologia , Inflamação/veterinária , Proteínas Quinases JNK Ativadas por Mitógeno/genética , MicroRNAs/genética , Receptores CCR7/genética , Aeromonas hydrophila/fisiologia , Animais , Sequência de Bases , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Inflamação/genética , Inflamação/imunologia , Inflamação/microbiologia , Proteínas Quinases JNK Ativadas por Mitógeno/imunologia , Lipopolissacarídeos/farmacologia , MicroRNAs/imunologia , Receptores CCR7/imunologia
7.
Fish Shellfish Immunol ; 94: 548-557, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31539573

RESUMO

A 56-day growth trial was conducted to investigate the effects of dietary yeast hydrolysate on the growth performance, antioxidation, immune response and resistance against Aeromonas hydrophila in largemouth bass. Four experimental diets were prepared with yeast hydrolysate levels of 0% (Y0), 1.5% (Y1.5), 3.0% (Y3.0) and 4.5% (Y4.5). Each diet was randomly assigned to triplicate 150-L tanks and each tank was stocked with 30 largemouth bass (initial body weight, IBW = 7.71 ±â€¯0.02 g). A challenge test was carried out after the feeding trial by injecting A. hydrophila intraperitoneally for 4-day observation. The results showed that the FBW and WGR in Y1.5 group were significantly higher than those in Y0 group (P < 0.05) and the feed conversion ratio (FCR) got the lowest value in Y1.5 group. And the hydrolysate supplement significantly increased the 4-day cumulative survival rate after the bacterial challenge (P < 0.05). The plasma malondialdehyde was lower in the yeast hydrolysate supplement groups in both pre- and post-challenge test (P < 0.05), while the plasma C3 increased (P < 0.05). In post-challenge test, the plasma superoxide dismutase (SOD) and catalase (CAT) activities increased in the Y1.5 and Y3.0 groups respectively (P < 0.05), and plasma lysozyme in Y1.5 group and the plasma IgM in Y3.0 group were higher than those in others respectively (P < 0.05). For the q-PCR results, in post-challenge test, the hepatic hep2 expression level in Y1.5 and Y4.5 groups were both significantly higher than those in others (P < 0.05), as well as il-8 in Y3.0 group. The spleen hif-1alpha and tgf-beta1 expression levels in Y4.5 group were all significantly lower than those in others (P < 0.05), while the gilt was significantly higher (P < 0.05) in the post-challenge test. And the expression levels of spleen tnf-alpah1 in Y1.5 and Y3.0 groups and il-8 in Y3.0 group were all significantly higher than those in other groups (P < 0.05) in the post-challenge test. The head kidney gilt expression level was significantly higher in the yeast hydrolysate supplement groups compared with the Y0 group (P < 0.05), and the head kidney il-8 expression level in Y1.5 group was significant higher than those in other groups in post-challenge test (P < 0.05). The present results indicated dietary yeast hydrolysate improved the antioxidant ability and enhanced the immune response of largemouth bass without negative effect on growth. And 1.5% or 3.0% of dietary yeast hydrolysate was recommended for largemouth bass based on the present results.


Assuntos
Bass , Resistência à Doença/efeitos dos fármacos , Doenças dos Peixes/imunologia , Expressão Gênica/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Fermento Seco/metabolismo , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Bass/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Doenças dos Peixes/genética , Doenças dos Peixes/metabolismo , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/metabolismo , Fermento Seco/administração & dosagem
8.
J Microbiol Biotechnol ; 29(8): 1324-1334, 2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31370117

RESUMO

Fish mycobacteriosis is a common bacterial disease in many species of freshwater and marine fish and has caused severe loss of fish production. Mycobacterium marinum has been the most prevalent pathogen observed in several outbreaks of mycobacteriosis of farmed sturgeons in China. However, the immune responses and pathology of sturgeons in mycobacterial infection are rarely studied. Therefore, we used the Illumina RNA-seq method to analyze the transcriptome profile of Acipenser schrenckii challenged with Mycobacterium marinum. To begin, 168,220 non-redundant contigs were acquired from the infection and control groups, and among these, 33,225 contigs have acquired annotations. A total of 4,043 differently expressed (DE) contigs between the two groups were identified, and among these, 2479 were upregulated and 1564 were down-regulated in the infected fish. A total of 1,340 DE contigs with acquired annotations in KEGG were enriched for 124 pathways including the TNF signaling pathway, and the Toll-like receptor signaling pathway. The roles of DE genes involved in significant pathways and other processes were discussed. The 2,209 DE contigs that have yet to acquire proper annotation may represent candidate genes associated with infection in sturgeons and are expected to serve as immunogenetic resources for further study. To our best knowledge, this is the first transcriptome study on sturgeons under bacterial infection.


Assuntos
Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Peixes/genética , Infecções por Mycobacterium não Tuberculosas/genética , Infecções por Mycobacterium não Tuberculosas/imunologia , Infecções por Mycobacterium não Tuberculosas/veterinária , Transcriptoma , Animais , China , Regulação para Baixo , Doenças dos Peixes/microbiologia , Proteínas de Peixes/metabolismo , Perfilação da Expressão Gênica , Imunidade , Anotação de Sequência Molecular , Mycobacterium marinum/patogenicidade , Regulação para Cima
9.
J Fish Dis ; 42(10): 1409-1417, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31424570

RESUMO

Bacillary necrosis of Pangasius (BNP), caused by Edwardsiella ictaluri, is one of the most devastating diseases in striped catfish farming. To date, quantitative genetic inheritance of BNP resistance is not known in striped catfish Pangasianodon hypophthalmus. The main aim of this study was to estimate genetic parameters for BNP resistance in a breeding population of striped catfish undergoing four generations of selection for high growth. Specifically, the study examined whether BNP resistance is heritable to enable family selection and whether genetic improvement for enhanced BNP resistance may have detrimental effects on growth and survival rate. To test these hypotheses, 720 full- and half-sib families were challenged with E. ictaluri pathogen using injection and cohabitation methods over four years, from 2010 to 2012 and 2015. In total, the data included 398,234 animals in the pedigree, from which 18,849 animals had disease challenge test records and 39,103 siblings had growth performance. Both univariate and bivariate sire-dam linear and threshold mixed models were used to estimate (co)variance components for BNP resistance, survivals and growth traits. The estimates of heritability for the BNP resistance recorded as death or survival were low regardless of models used (0.10-0.16), whereas survival time (days post-challenge test) showed moderate heritability (0.35). The survival rate during hapa rearing had medium heritability (0.33-0.52). The genetic correlations of BNP resistance with body weight and survival were all positive (0.03-0.53), suggesting that selection of increased BNP resistance may have positive impacts on growth and survival traits, and these traits could be easily improved simultaneously in the selective breeding programme for striped catfish.


Assuntos
Cruzamento , Peixes-Gato , Resistência à Doença/genética , Infecções por Enterobacteriaceae/veterinária , Doenças dos Peixes/genética , Animais , Edwardsiella ictaluri/fisiologia , Infecções por Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/microbiologia , Doenças dos Peixes/microbiologia
10.
Fish Shellfish Immunol ; 94: 58-65, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31470137

RESUMO

TIR domain-containing protein is an important member for some bacterial pathogens to subvert host defenses. Here we described a fish virulent Yersinia ruckeri SC09 strain that interfered directly with Toll-like receptor (TLR) function by a TIR-containing protein. Firstly, the novel TIR-containing protein was identified by bioinformatics analysis and named as TcpA. Secondly, the toxic effects of TcpA in fish was demonstrated in vivo challenge experiments through knockout mutant and complement mutant of tcpA gene. Thirdly, The study in vitro revealed that TcpA could down-regulate the expression and secretion of IL-6, IL-1ß and TNF-α. Finally, we demonstrated that TcpA could inhibit the TLR signaling pathway through interaction with myeloid differentiation factor 88 (MyD88) in experiments such as NF-κB dependent luciferase reporter system, co-immunoprecipitation, GST pull-down and yeast two-hybrid. The study revealed that TcpA was essential for virulence and was able to interact with the TIR adaptor protein MyD88 and inhibit the pre-inflammatory signal of immune cells and promote the intracellular survival of pathogenic Yersinia ruckeri SC09 strain. In conclusion, our results showed that TcpA acted as a new virulence factor in Y. ruckeri could suppress innate immune response and increase virulence by inhibiting TLR and MyD88-mediated specific signaling, highlighting a novel strategy for innate immune evasion in bacteria.


Assuntos
Evasão da Resposta Imune/genética , Imunidade Inata/genética , Fator 88 de Diferenciação Mieloide/genética , Receptores Toll-Like/genética , Fatores de Virulência/genética , Yersiniose/veterinária , Yersinia ruckeri/genética , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Fator 88 de Diferenciação Mieloide/imunologia , Transdução de Sinais/imunologia , Receptores Toll-Like/metabolismo , Fatores de Virulência/metabolismo , Yersiniose/genética , Yersiniose/imunologia
11.
Fish Shellfish Immunol ; 94: 249-257, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31470139

RESUMO

In recent years, streptococcal diseases have severely threatened the development of tilapia aquaculture, but effective prevention and control methods have not yet been established. To understand the immune responses of vaccinated Nile tilapia (Oreochromis niloticus), digital gene expression (DGE) technology was applied in this study to detect the gene expression profile of the Nile tilapia (O. niloticus) liver in response to ScpB (Streptococcal C5a peptidase from group B Streptococcus, ScpB) vaccination and a Streptococcus agalactiae-challenge. The control and the ScpB-vaccinated Nile tilapia yielded a total of 25,788,734 and 27,088,598 clean reads, respectively. A total of 1234 significant differentially expressed unigenes were detected (P < 0.05), of which 236 were significantly up-regulated, and 269 were significantly down-regulated (P < 0.05, |fold|>2, FDR<0.05). Of the differentially expressed gene, the identified genes which were enriched using databases of GO and KEGG could be categorized into a total of 67 functional groups and were mapped to 153 signaling pathways including 15 immune-related pathways. The differentially expressed genes (TLR1, TLR2, TLR3, TLR5, TLR9, MyD88, C3, IL-1ß, IL-10) were detected in the expression profiles, and this was subsequently verified via quantitative real-time PCR (qPCR). The results of this study can serve as a basis for future research not only on the molecular mechanism of S. agalactiae invasion, but also on the anti-S. agalactiae mechanism in targeted tissues of Nile tilapia.


Assuntos
Ciclídeos/imunologia , Doenças dos Peixes/genética , Perfilação da Expressão Gênica/veterinária , Regulação da Expressão Gênica , Infecções Estreptocócicas/veterinária , Streptococcus agalactiae/fisiologia , Animais , Ciclídeos/genética , Regulação para Baixo , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Biblioteca Gênica , Ontologia Genética , Fígado/metabolismo , Fígado/microbiologia , Infecções Estreptocócicas/genética , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/microbiologia , Vacinas Estreptocócicas/administração & dosagem , Regulação para Cima
12.
Genet Sel Evol ; 51(1): 42, 2019 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-31387519

RESUMO

BACKGROUND: Columnaris disease (CD) is an emerging problem for the rainbow trout aquaculture industry in the US. The objectives of this study were to: (1) identify common genomic regions that explain a large proportion of the additive genetic variance for resistance to CD in two rainbow trout (Oncorhynchus mykiss) populations; and (2) estimate the gains in prediction accuracy when genomic information is used to evaluate the genetic potential of survival to columnaris infection in each population. METHODS: Two aquaculture populations were investigated: the National Center for Cool and Cold Water Aquaculture (NCCCWA) odd-year line and the Troutlodge, Inc., May odd-year (TLUM) nucleus breeding population. Fish that survived to 21 days post-immersion challenge were recorded as resistant. Single nucleotide polymorphism (SNP) genotypes were available for 1185 and 1137 fish from NCCCWA and TLUM, respectively. SNP effects and variances were estimated using the weighted single-step genomic best linear unbiased prediction (BLUP) for genome-wide association. Genomic regions that explained more than 1% of the additive genetic variance were considered to be associated with resistance to CD. Predictive ability was calculated in a fivefold cross-validation scheme and using a linear regression method. RESULTS: Validation on adjusted phenotypes provided a prediction accuracy close to zero, due to the binary nature of the trait. Using breeding values computed from the complete data as benchmark improved prediction accuracy of genomic models by about 40% compared to the pedigree-based BLUP. Fourteen windows located on six chromosomes were associated with resistance to CD in the NCCCWA population, of which two windows on chromosome Omy 17 jointly explained more than 10% of the additive genetic variance. Twenty-six windows located on 13 chromosomes were associated with resistance to CD in the TLUM population. Only four associated genomic regions overlapped with quantitative trait loci (QTL) between both populations. CONCLUSIONS: Our results suggest that genome-wide selection for resistance to CD in rainbow trout has greater potential than selection for a few target genomic regions that were found to be associated to resistance to CD due to the polygenic architecture of this trait, and because the QTL associated with resistance to CD are not sufficiently informative for selection decisions across populations.


Assuntos
Cruzamento , Mapeamento Cromossômico , Doenças dos Peixes/genética , Infecções por Flavobacteriaceae/veterinária , Flavobacterium , Oncorhynchus mykiss/genética , Animais , Resistência à Doença/genética , Feminino , Pesqueiros , Infecções por Flavobacteriaceae/genética , Padrões de Herança , Masculino , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Seleção Genética
13.
Fish Shellfish Immunol ; 93: 1041-1046, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31465870

RESUMO

Although the type I interferon-mediated increase of Mx1 and ISG15 gene expression in Epithelioma papulosum cyprini (EPC) cells has been reported, the antiviral role of Mx1 and ISG15 in EPC cells has not been investigated. In this study, to know the anti-viral hemorrhagic septicemia virus (VHSV) role of Mx1 and ISG15 of EPC cells, either Mx1 or ISG15 gene was knocked-out using a CRISPR/Cas9 system, and the progression of cytopathic effects (CPE) and viral growth were analyzed. Mx1 gene and ISG15 gene knockout EPC cells were successfully produced via CRISPR/Cas9 coupled with a single-cell cloning. Through the sequence analysis, one clone showing two heterozygous indel patterns in Mx1 gene and a clone showing three heterozygous indel patterns in ISG15 gene were selected for further analyses. Mx1 knockout EPC cells did not show any differences in VHSV-mediated CPE progression, even when pre-treated with polyinosinic:polycytidylic acid (poly I:C), compared to control EPC cells. These results suggest that Mx1 in EPC cells may be unfunctional to cytoplasmic RNA viruses. In contrast to Mx1, ISG15 knockout cells showed clearly hampered anti-VHSV activity even when pre-treated with poly I:C, indicating that ISG15 plays an important role in type I interferon-mediated anti-viral activity in EPC cells, which allowed VHSV to replicate more efficiently in ISG15 knockout cells than Mx1 knockout and control cells.


Assuntos
Sistemas CRISPR-Cas/imunologia , Cyprinidae/imunologia , Resistência à Doença/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Expressão Gênica , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular Tumoral , Cyprinidae/genética , Resistência à Doença/imunologia , Doenças dos Peixes/genética , Técnicas de Inativação de Genes/veterinária , Septicemia Hemorrágica Viral/genética , Septicemia Hemorrágica Viral/imunologia , Interferon Tipo I/genética , Novirhabdovirus/fisiologia , Poli I-C/farmacologia , Análise de Sequência de DNA/veterinária , Análise de Sequência de Proteína/veterinária
14.
Genet Sel Evol ; 51(1): 47, 2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31455244

RESUMO

BACKGROUND: Infectious hematopoietic necrosis (IHN) is a disease of salmonid fish that is caused by the IHN virus (IHNV). Under intensive aquaculture conditions, IHNV can cause significant mortality and economic losses. Currently, there is no proven and cost-effective method for IHNV control. Clear Springs Foods, Inc. has been applying selective breeding to improve genetic resistance to IHNV in their rainbow trout breeding program. The goals of this study were to elucidate the genetic architecture of IHNV resistance in this commercial population by performing genome-wide association studies (GWAS) with multiple regression single-step methods and to assess if genomic selection can improve the accuracy of genetic merit predictions over conventional pedigree-based best linear unbiased prediction (PBLUP) using cross-validation analysis. RESULTS: Ten moderate-effect quantitative trait loci (QTL) associated with resistance to IHNV that jointly explained up to 42% of the additive genetic variance were detected in our GWAS. Only three of the 10 QTL were detected by both single-step Bayesian multiple regression (ssBMR) and weighted single-step GBLUP (wssGBLUP) methods. The accuracy of breeding value predictions with wssGBLUP (0.33-0.39) was substantially better than with PBLUP (0.13-0.24). CONCLUSIONS: Our comprehensive genome-wide scan for QTL revealed that genetic resistance to IHNV is controlled by the oligogenic inheritance of up to 10 moderate-effect QTL and many small-effect loci in this commercial rainbow trout breeding population. Taken together, our results suggest that whole genome-enabled selection models will be more effective than the conventional pedigree-based method for breeding value estimation or the marker-assisted selection approach for improving the genetic resistance of rainbow trout to IHNV in this population.


Assuntos
Doenças dos Peixes/genética , Vírus da Necrose Hematopoética Infecciosa , Oncorhynchus mykiss/genética , Infecções por Rhabdoviridae/veterinária , Animais , Teorema de Bayes , Cruzamento , Cruzamentos Genéticos , Resistência à Doença/genética , Doenças dos Peixes/virologia , Pesqueiros , Estudo de Associação Genômica Ampla/veterinária , Herança Multifatorial , Oncorhynchus mykiss/virologia , Locos de Características Quantitativas , Infecções por Rhabdoviridae/genética
15.
J Fish Dis ; 42(11): 1553-1562, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31448825

RESUMO

The potential of selection to improve resistance to streptococcosis was evaluated in a commercial population of Nile tilapia in Thailand. The base generation (G0) consisted of offspring from 98 sires and 149 dams using a partly nested design. At 60 days post-hatch, 30 fish from each family were injected intraperitoneally with a Streptococcosis agalactiae solution (1 × 109  CFU/ml) and evaluated for 14 days. Disease resistance was recorded as the number of days from challenge until death (DD) and as a binary (BIN) trait (dead/alive) on day 14. Three models were used for genetic analyses: Cox frailty model for DD; animal model for DD; and animal model for BIN. Age at challenge was fitted as a covariate and contemporary group as fixed or random effect, depending on the model. Fish from the 18 most resistant families were selected to produce the first generation (G1). Heritability estimates for G0 were 0.22, 0.14 ± 0.02 and 0.11 ± 0.02 for the Cox, linear DD and linear BIN models, respectively. Selection response indicated that the risk of death decreased to 54%, survival time increased to 3.4 days and survival rate increased to 21%. These results suggest that genetic improvement is possible for this population.


Assuntos
Ciclídeos , Resistência à Doença , Doenças dos Peixes/genética , Seleção Genética , Infecções Estreptocócicas/veterinária , Streptococcus agalactiae/fisiologia , Animais , Doenças dos Peixes/microbiologia , Modelos Biológicos , Infecções Estreptocócicas/genética , Infecções Estreptocócicas/microbiologia
16.
Biomed Res Int ; 2019: 9617659, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31317044

RESUMO

The yellow catfish (Pelteobagrus fulvidraco) is an important economic freshwater aquaculture species in Asia. However, little is known about its immune response to bacterial pathogen infection. Here, two cytokines, the proinflammatory cytokine interleukin-8 (IL-8) and the anti-inflammatory cytokine interleukin-10 (IL-10), were identified and characterized in the yellow catfish for the first time. We found that the full length of the IL-8 cDNA was 784 bp and contained an open reading frame (ORF) of 336 bp, while the IL-10 gene was 973 bp in length with a 549 bp of ORF. In addition, both the IL-8 and the IL-10 had similar tissue-specific expression patterns. They were more abundant in the spleen and lowest expressed in the liver. Furthermore, IL-10 but not IL-8 was significantly upregulated in the intestine of yellow catfish by feed supplementation of Clostridium butyricum (CB). More importantly, the expression levels of intestinal IL-10 and IL-8 were up- and downregulated by pathogen Aeromonas punctata stimuli with the presence of CB, respectively. Collectively, these results suggest that IL-10 and IL-8 mediate important roles in the immunity of yellow catfish, and feed supplementation of CB may able to reduce the intestinal inflammation caused by bacteria infections through regulating the expression of IL-10 and IL-8.


Assuntos
Infecções Bacterianas/genética , Peixes-Gato/genética , Interleucina-10/genética , Interleucina-8/genética , Animais , Infecções Bacterianas/microbiologia , Infecções Bacterianas/patologia , Peixes-Gato/microbiologia , Clonagem Molecular , Doenças dos Peixes/genética , Doenças dos Peixes/microbiologia , Regulação da Expressão Gênica/genética , Fígado/microbiologia , Filogenia , Baço/microbiologia
17.
Fish Shellfish Immunol ; 93: 99-107, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31323328

RESUMO

Epinephelus moara is an economically important fish in Southeast Asian countries but is suffering from nervous necrosis virus (NNV) infection. A deeper understanding of the host-NNV interaction mechanisms makes sense for disease control, however, at present, the pathogenesis of natural NNV infection and the resistance mechanism in host remains poorly understood. In this study, asymptomatic and diseased E. moara with clinical symptoms of viral nervous necrosis (VNN) from a grouper farm were both detected with a positive RT-PCR signal of NNV, then transcriptome sequencing of their immune tissues (liver, spleen and kidney) were performed for comparation analysis. The de novo assemblies yielded 53,789 unigenes which had a length varied from 201 to 19,675 bp and a N50 length of 2115 bp, and 29,451 unigenes were functionally annotated, with 83, 250 and 5632 unigenes being differentially expressed in liver, spleen and kidney respectively. KEGG pathway enrichment analysis of the DEGs showed many DEGs were enriched in immune related pathways. Although the expression of class I major histocompatibility complex (MHC) was significantly higher in three immune tissues of the diseased grouper, many immune related genes, including humoral immune molecules (such as antibodies), the cellular mediated cytotoxic molecules (such as perforin) and some adhesion related genes were down regulated in the diseased grouper. Our results provided many unigenes that might play important roles in NNV resistance for further research. Furthermore, a total of 8666 unigenes containing 11,623 simple sequence repeats (SSRs) were identified, which provided useful information for screening molecular markers associated with NNV resistance in E. moara.


Assuntos
Bass/genética , Bass/imunologia , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Infecções por Vírus de RNA/veterinária , Transcriptoma/imunologia , Animais , Doenças dos Peixes/genética , Perfilação da Expressão Gênica/veterinária , Nodaviridae/fisiologia , Infecções por Vírus de RNA/genética , Infecções por Vírus de RNA/imunologia
18.
Fish Shellfish Immunol ; 93: 269-277, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31306762

RESUMO

As an isoform of Rho family GTPases, RhoB plays a pivotal role in cytoskeletal organization, cell proliferation, apoptosis and immune response. However, the regulatory mechanisms of RhoB expression in aquatic animals are still unknown. In the present study, we first construct Vibrio anguillarum infection model in S. maximus, including susceptible and resistant individuals. Then the temporal expression of RhoB was detected after V. anguillarum challenge using qRT-PCR and found that RhoB transcripts were significantly induced in the liver, gill and blood despite of differential expression levels and responsive time points. In addition, the mRNA levels of RhoB in resistant individuals were significantly higher than in susceptible ones. The length of 2083 bp sequences of RhoB promoter was cloned and characterized. Moreover, DNA methylation of the RhoB promoter was measured by bisulfite sequencing (BSP) and hypo-methylated was detected in the CpG islands. Three SNPs (-1590, -1575 and -1449) and two haplotypes in the promoter region of RhoB were identified to be associated with V. anguillarum resistance in turbot by association analysis in group 17-R and 17-S. Deletion analysis indicated that these SNPs could negatively mediate the activity of RhoB promoter. Site-directed mutagenesis and qRT-PCR of individuals with different genotypes demonstrated that -1575 T/A polymorphism affected promoter activity. Further study showed that this mutation altered the binding site of the transcription factor CREB. Co-transfection of SmCREB and RhoB promoter was performed in HEK293T cells which confirmed the -1575 allelic differences on transcriptional activity, with the susceptibility allele showing reduced activity. Taken together, our findings implicate that losing of binding of CREB to SmRhoB promoter due to -1575T/A polymorphisms enhances SmRhoB expression in resistant turbot, which provide insights into the effect of SmRhoB expression in response to V. anguillarum infection.


Assuntos
Doenças dos Peixes/imunologia , Linguados/genética , Polimorfismo de Nucleotídeo Único/imunologia , Vibrio/fisiologia , Proteína rhoB de Ligação ao GTP/imunologia , Animais , Proteína de Ligação a CREB/genética , Proteína de Ligação a CREB/metabolismo , Suscetibilidade a Doenças/imunologia , Suscetibilidade a Doenças/veterinária , Doenças dos Peixes/genética , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Linguados/imunologia , Regulação da Expressão Gênica/imunologia , Haplótipos/imunologia , Mutação , Regiões Promotoras Genéticas/genética , Regiões Promotoras Genéticas/imunologia , Vibrioses/imunologia , Vibrioses/veterinária , Proteína rhoB de Ligação ao GTP/genética
19.
Fish Shellfish Immunol ; 92: 209-215, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31181340

RESUMO

Disease outbreak is a major obstruction for intensive aquaculture worldwide. One of the promising solutions is genetic improvement by selective breeding, providing that a sufficient proportion of additive genetic variance (measured by heritability-h2) of disease resistance traits exists. In addition, immunity traits are of interest as potential indirect targeted traits for disease resistance. In this study, the genetic parameters of resistance to Aeromonas hydrophila were reported for the first time in the bighead catfish, Clarias macrocephalus Günther, 1864 which is an important parental species for the production of the commercially important hybrid C. macrocephalus × C. gariepinus. The analyses were performed on 736 data records obtained from 74 full-sib families (31 half-sib families) produced by factorial mating design. The results showed that the heritability of survival rate after disease (Aeromonas hydrophila) challenge (intraperitoneal injection with 0.1 ml containing 1 × 106 CFU/ml of A. hydrophila) was low to moderate (0.05 ±â€¯0.02-0.27 ±â€¯0.15). The immune traits (bactericidal activity-BA, lysozyme activity-LA, and alternative complement activity-ACH50) had low to moderate heritability (h2BA = 0.05 ±â€¯0.02; h2LA = 0.16 ±â€¯0.04; h2ACH50 = 0.31 ±â€¯0.06) while heritability of hematocrit (Hct) was also low (h2Hct = 0.17 ±â€¯0.04). The results suggested the possibility to improve resistance to A. hydrophila by selection, while the possibility to use immunity traits as indirect selection criteria for disease resistance is still unclear.


Assuntos
Peixes-Gato/genética , Doenças dos Peixes/imunologia , Hereditariedade/imunologia , Imunidade Inata/genética , Aeromonas hydrophila/fisiologia , Animais , Peixes-Gato/imunologia , Resistência à Doença/genética , Doenças dos Peixes/genética , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária
20.
Artigo em Inglês | MEDLINE | ID: mdl-31174158

RESUMO

Flavobacterium psychrophilum is the etiologic agent of rainbow trout fry syndrome (RTFS). This pathogen infects a wide variety of salmonid species during freshwater stages, causing significant losses in the aquaculture industry. Rainbow trout (Oncorhynchus mykiss) infected with F. psychrophilum, presents as the main external clinical sign ulcerative lesions and necrotic myositis in skeletal muscle. We previously reported the in vitro cytotoxic activity of F. psychrophilum on rainbow trout myoblast, however little is known about the molecular mechanisms underlying the in vivo pathogenesis in skeletal muscle. In this study, we examined the transcriptomic profiles of skeletal muscle tissue of rainbow trout intraperitoneally challenged with low infection dose of F. psychrophilum. Using high-throughput RNA-seq, we found that 233 transcripts were up-regulated, mostly associated to ubiquitin mediated proteolysis and apoptosis. Conversely, 189 transcripts were down-regulated, associated to skeletal muscle contraction. This molecular signature was consistent with creatine kinase activity in plasma and oxidative damage in skeletal muscle. Moreover, the increased caspase activity suggests as a whole skeletal muscle atrophy induced by F. psychrophilum. This study offers an integrative analysis of the skeletal muscle response to F. psychrophilum infection and reveals unknown aspects of its pathogenesis in rainbow trout.


Assuntos
Doenças dos Peixes/genética , Infecções por Flavobacteriaceae/veterinária , Flavobacterium/fisiologia , Oncorhynchus mykiss/genética , Transcriptoma , Animais , Aquicultura , Doenças dos Peixes/microbiologia , Doenças dos Peixes/patologia , Infecções por Flavobacteriaceae/genética , Infecções por Flavobacteriaceae/microbiologia , Infecções por Flavobacteriaceae/patologia , Interações Hospedeiro-Patógeno , Músculo Esquelético/metabolismo , Músculo Esquelético/microbiologia , Músculo Esquelético/patologia , Oncorhynchus mykiss/microbiologia
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