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1.
Virology ; 592: 110008, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38335866

RESUMO

Viral spike proteins undergo a special maturation process that enables host cell receptor recognition, membrane fusion, and viral entry, facilitating effective virus infection. Here, we investigated the protease cleavage features of ORF46, a spike-like protein in Ictalurid herpesvirus 1 (IcHV-1) sharing similarity with spikes of Nidovirales members. We noted that during cleavage, full-length ORF46 is cleaved into ∼55-kDa and ∼100-kDa subunits. Moreover, truncation or site-directed mutagenesis at the recognition sites of proprotein convertases (PCs) abolishes this spike cleavage, highlighting the crucial role of Arg506/Arg507 and Arg668/Arg671 for the cleavage modification. ORF46 cleavage was suppressed by specific N-glycosylation inhibitors or mutation of its specific N-glycosylation sites (N192, etc.), suggesting that glycoprotein ORF46 cleavage is modulated by N-glycosylation. Notably, PCs and N-glycosylation inhibitors exhibited potent antiviral effects in host cells. Our findings, therefore, suggested that PCs cleavage of ORF46, modulated by N-glycosylation, is a potent antiviral target for fish herpesviruses.


Assuntos
Ictalurivirus , Pró-Proteína Convertases , Animais , Pró-Proteína Convertases/genética , Pró-Proteína Convertases/metabolismo , Glicosilação , Proteínas Virais/genética , Proteínas Virais/metabolismo , Antivirais
2.
Fish Shellfish Immunol ; 141: 109051, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37689228

RESUMO

Herein, the effects of Agaricus bisporus Polysaccharides (ABPs) on anti-channel catfish virus (CCV) infections to promote their application in channel catfish culture were explored. Transcriptome and metabolome analyses were conducted on the spleen of a CCV-infected channel catfish model fed with or without ABPs. CCV infections upregulated many immune and apoptosis-related genes, such as IL-6, IFN-α3, IFN-γ1, IL-26, Casp3, Casp8, and IL-10, and activated specific immunity mediated by B cells. However, after adding ABPs, the expression of inflammation-related genes decreased in CCV-infected channel catfish, and the inflammatory inhibitors NLRC3 were upregulated. Meanwhile, the expression of apoptosis-related genes was reduced, indicating that ABPs can more rapidly and strongly enhance the immunity of channel catfish to resist viral infection. Moreover, the metabonomic analysis showed that channel catfish had a high energy requirement during CCV infection, and ABPs could enhance the immune function of channel catfish. In conclusion, ABPs can enhance the antiviral ability of channel catfish by enhancing immune response and regulating inflammation. Thus, these findings provided new insights into the antiviral response effects of ABPs, which might support their application in aquaculture.


Assuntos
Doenças dos Peixes , Ictaluridae , Ictalurivirus , Animais , Imunidade , Inflamação , Antivirais
3.
Viruses ; 15(8)2023 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-37632031

RESUMO

The efficacy of silver nanoparticles (AgNPs) was tested in vitro against three different fish viruses, causing significant economic damage in aquaculture. These viruses were the spring viraemia of carp virus (SVCV), European catfish virus (ECV), and Ictalurid herpesvirus 2 (IcHV-2). The safe concentration of AgNPs that did not cause cytotoxic effects in EPC cells proved to be 25 ng/mL. This dose of AgNPs decreased significantly (5-330×) the viral load of all three viruses in three different types of treatments (virus pre-treatment, cell pre-treatment, and cell post-treatment with the AgNPs). In a higher concentration, the AgNPs proved to be efficient against ECV and IcHV-2 even in a delayed post-cell-treatment experiment (AgNP treatment was applied 24 h after the virus inoculation). These first in vitro results against three devastating fish viruses are encouraging to continue the study of the applicability of AgNPs in aquaculture in the future.


Assuntos
Peixes-Gato , Ictalurivirus , Nanopartículas Metálicas , Animais , Antivirais/farmacologia , Prata/farmacologia
4.
Virus Res ; 334: 199163, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37364814

RESUMO

The family Alloherpesviridae contains herpesviruses of fish and amphibians. Due to the significant economic losses to aquaculture that herpesviruses can cause, the primary areas of research interest are concerning their pathogenesis and prevention. Despite alloherpesvirus genomic sequences becoming more widely accessible, methods regarding their genus/species classification are still relatively unexplored. In the present study, the phylogenetic relationships between 40 completely sequenced alloherpesviruses were illustrated by the viral proteomic tree (ViPTree), which was divided into three monophyletic groups, namely Cyprinivirus, Ictalurivirus and Batrachovirus. Additionally, average nucleotide identity (ANI) and average amino acid identity (AAI) analyses were performed across all available sequences and clearly displayed species boundaries with the threshold value of ANI/AAI set at 90%. Subsequently, core-pan analysis uncovered 809 orthogroups and 11 core genes shared by all 40 alloherpesvirus genome sequences. For the former, a 15 percent identity depicts a clear genus boundary; for the latter, 8 of them may be qualified for phylogenetic analysis based on amino acid or nucleic acid sequences after being verified using maximum likelihood (ML) or neighbor-joining (NJ) phylogenetic trees. Finally, although the dot plot analysis was valid for the members within Ictalurivirus, it was unsuccessful for Cyprinivirus and Batrachovirus. Taken together, the comparison of individual methodologies provides a wide range of alternatives for alloherpesviruses classification under various circumstances.


Assuntos
Herpesviridae , Ictalurivirus , Animais , Filogenia , Proteômica , Herpesviridae/genética , Ictalurivirus/genética , Genômica/métodos
5.
Virus Res ; 315: 198794, 2022 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-35504448

RESUMO

Channel catfish virus (CCV), an important member of the family Alloherpesviridae, causes a lethal infection in channel catfish. As with most animal viruses, the initial step of infection by CCV is entry into host cells, which is also a promising antiviral target for CCV disease. This study investigated the mechanism of host cell invasion by CCV using a series of biochemical inhibitor assays in channel catfish cells. CCV infection in host cells was does-dependently inhibited when cells were treated with endosomal acidification inhibitors (5 µM chloroquine, 50 nM bafilomycin A1, and 1 mM ammonium chloride) and hypertonic medium (50 mM sucrose) , which suggests that CCV invades host cells in a manner dependent on low-pH and the endocytic pathway. Moreover, when the cells were pretreated with inhibitors of clathrin-mediated endocytosis, including chlorpromazine (2 µM) and dynasore (50 µM), the CCV infection in the host cells was strongly inhibited. In contrast, the destruction of cellular cholesterol by methyl-ß-cyclodextrin and nystatin and inhibition of macropinocytosis had no effect on viral entry. Altogether, these findings indicate that CCV infects host cells via clathrin-mediated endocytosis in a low-pH-dependent manner, suggesting that this CCV entry pathway offers an antiviral target against CCV disease.


Assuntos
Clatrina , Ictalurivirus , Animais , Antivirais/farmacologia , Linhagem Celular , Clatrina/metabolismo , Endocitose , Internalização do Vírus
6.
J Fish Dis ; 45(5): 655-666, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35176182

RESUMO

The channel catfish virus (CCV) is a lethal pathogen to aquatic animals that can provoke severe haemorrhagic disease in juvenile channel catfish. Although the CCV genome has been fully sequenced, the molecular mechanisms of CCV infection and pathogenesis are less well known. Genomic DNA replication is a necessary and key event for the CCV life cycle. In this study, the impacts of the putative helicase and primase encoded by viral ORF25 and ORF63 on CCV genome replication and infection were evaluated in channel catfish ovary (CCO) cells. The results showed that the number of CCV genome copies was decreased significantly in virus-infected CCO cells after knockdown of ORF25 and ORF63 using RNA interference. In contrast, the overexpression of ORF25 and ORF63 led to slight increase in the number of virus genome copies. Consistent with the above results, the present results also showed that the expressions of CCV true-late genes which strictly depend on viral DNA replication, were significantly increased or repressed by overexpression or RNA interference targeting viral ORF25 and ORF63 genes in virus-infected CCO cells. In addition, knockdown of ORF25 and ORF63 remarkably inhibited CCV-induced cytopathic effects and decreased progeny virus titres in CCO cells. Moreover, transmission electron microscopy observation of CCO cells infected with CCV accompanied by siRNA targeting the viral ORF25 and ORF63 genes showed that the number of virus particles was remarkably reduced. Taken together, these results indicated that ORF25 and ORF63 are essential for regulating CCV genome replication and CCV-induced infection. Our findings will provide an understanding of the replication mechanisms of CCV and contribute to the development of antiviral strategies for controlling CCV infection in channel catfish culture.


Assuntos
Doenças dos Peixes , Ictaluridae , Ictalurivirus , Animais , Replicação do DNA , DNA Viral/genética , Feminino , Ictaluridae/genética , Ictalurivirus/genética , Replicação Viral
7.
Viruses ; 13(12)2021 11 29.
Artigo em Inglês | MEDLINE | ID: mdl-34960662

RESUMO

The channel catfish virus (CCV, Ictalurid herpesvirus 1) has caused sustained economic losses in the fish industry because of its strong infectivity and pathogenicity. Thus, it is necessary to determine the function of viral proteins in the CCV infection process. The present study aimed to characterize CCV glycoprotein ORF59 and explore its impact on virus infection in host cells. Firstly, its exclusive presence in the membrane fraction of the cell lysate and subcellular localization verified that CCV ORF59 is a viral membrane protein expressed at late-stage infection. A protein blocking assay using purified His6 tagged ORF59, expressed in sf9 insect cells using a baculovirus expression system, indicated a dose-dependent inhibitory effect of recombinant ORF59 protein on virus invasion. Knockdown of the ORF59 using a short hairpin (shRNA) showed that ORF59 silencing decreased the production of infectious virus particles in channel catfish ovary cells. The results of this study suggest that recombinant ORF59 protein might inhibit CCV entry into the host cells. These findings will promote future studies of the key functions of glycoprotein ORF59 during CCV infection.


Assuntos
Doenças dos Peixes/virologia , Glicoproteínas/metabolismo , Infecções por Herpesviridae/veterinária , Ictalurivirus/fisiologia , Proteínas Virais/metabolismo , Internalização do Vírus , Animais , Peixes-Gato/virologia , Glicoproteínas/genética , Infecções por Herpesviridae/virologia , Ictalurivirus/genética , Proteínas Virais/genética , Replicação Viral
8.
J Fish Dis ; 44(10): 1609-1617, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34192354

RESUMO

The expression of herpesvirus genes during infection of tissue culture cells can be classified into three main classes: immediate-early (IE), early and late. The transcriptional regulation of herpesvirus IE genes is a critical regulatory step in the initiation of viral infection, with their regulation differing from that of early and late genes. Herein, we report that an IE gene (ORF3) promoter in channel catfish virus (CCV, Ictalurid herpesvirus 1) can be activated regardless of the presence or absence of CCV infection, indicating that the ORF3 promoter is efficiently driven by host-cell transcription factors in a viral infection-independent manner. The analysis of truncated promoter activity suggested that several transcription elements play a role in activating the ORF3 promoter, with the key cis-elements seemingly located in the flanking sequence of the start codon ATG. We further found that this flanking sequence contained multiple AT-rich sequences, and systematic mutational analyses showed that these AT-rich sequences affected normal transcription levels of the ORF3 promoter. To summarize, multiple AT-rich domains, representing the novel architecture of IE gene promoters in Ictalurid herpesvirus 1, serve as a cis-element for ORF3 transcription.


Assuntos
Doenças dos Peixes/virologia , Infecções por Herpesviridae/veterinária , Ictaluridae , Ictalurivirus/genética , Regiões Promotoras Genéticas , Proteínas Virais/genética , Animais , Linhagem Celular , Infecções por Herpesviridae/virologia
9.
J Fish Dis ; 44(9): 1399-1409, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34028055

RESUMO

Blue catfish alloherpesvirus (BCAHV) is a novel virus isolated from the blue catfish (Ictalurus furcatus). To date, the ultrastructure, virulence and immunogenicity of BCAHV have not been reported. Given the importance of blue catfish in producing channel ♀ (I. punctatus) × â™‚ blue (I. furcatus) catfish hybrids and the increasing demand for hybrid catfish in the US catfish industry, the susceptibility of blue, channel and hybrid catfish to BCAHV was assessed. Further, the cross-protective potential of BCAHV against Ictalurid herpesvirus 1 (IcHV1) was investigated in channel and hybrid catfish that survive BCAHV exposure. Neutralization assays revealed BCAHV is refractive (neutralization index [NI] = 0) to anti-IcHV1 monoclonal antibody Mab 95, compared to IcHV1 (NI = 1.8). Exposure of blue catfish fingerling to 1.3 × 105 TCID50 /L BCAHV produced cumulative mortality of 51.67 ± 0.70% and pathologic changes similar to those of channel catfish virus disease. No mortality was observed in channel or hybrid catfish. Twenty-eight days post-challenge, surviving channel and hybrid catfish were exposed to 9.4 × 104 TCID50 /L IcHV1 (LC50 dose), resulting in 100% relative per cent survival compared to naïve cohorts. These data provide baseline information for BCAHV and lay the groundwork for future studies. Data also identify BCAHV as a potential vaccine candidate against IcHV1. Based on host range and immunogenicity evaluations, in addition to genome sequence data from previous studies, BCAHV should be given consideration as a new species of Ictalurivirus.


Assuntos
Doenças dos Peixes/virologia , Infecções por Herpesviridae/veterinária , Ictalurivirus/patogenicidade , Animais , Suscetibilidade a Doenças/veterinária , Suscetibilidade a Doenças/virologia , Doenças dos Peixes/mortalidade , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/mortalidade , Ictaluridae , Ictalurivirus/imunologia , Virulência
10.
J AOAC Int ; 104(5): 1350-1354, 2021 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-33787893

RESUMO

BACKGROUND: Channel catfish virus disease (CCVD) has resulted in great economic losses and has restricted the development of fisheries. There is therefore, a need for rapid and efficient diagnostic methods to control the spread of CCVD. OBJECTIVE: A colloidal gold immunochromatographic strip has been developed for the detection of CCVD. METHODS: In this study, a colloidal gold immunochromatographic strip for channel catfish virus (CCV) detection was developed using the monoclonal antibody 8B6 conjugated with colloidal gold as the detector antibody. A rabbit anti-CCV antibody was used as the capture complex at the test line, and a goat anti-mouse IgG antibody was used as the capture antibody at the control line. The strip was characterized in its specificity, sensitivity, and stability. In addition, an infection experiment was performed to test the applicability of the test strip. RESULT: The strip was able to detect concentrations of the virus (104 tissue culture infective dose (TCID50)/mL) and showed analytical specificity when tested against other viral pathogens. The strips were still usable after 30 days of storage at 60°C. It was possible to detect CCV experimentally in infected fish within 10-15 min of using the strip. CONCLUSIONS: The strip can be used as a rapid and convenient tool for on-site diagnosis to control outbreaks and the spread of CCVD. HIGHLIGHTS: The immunochromatographic strip was the first to be developed and applied for the detection of CCVD.


Assuntos
Coloide de Ouro , Ictalurivirus , Animais , Anticorpos Monoclonais , Cromatografia de Afinidade , Camundongos , Coelhos , Sensibilidade e Especificidade
11.
Virus Res ; 292: 198249, 2021 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-33253717

RESUMO

The channel catfish virus (CCV) can cause lethal hemorrhagic infection in channel catfish, resulting in significant economic losses in the fish industry. Effective drugs for the virus are still lacking. Acyclovir is known as a potent antiviral agent against human herpes viruses and some animal DNA viruses. The present study was undertaken to explore the antiviral response and mechanism of acyclovir against CCV in channel catfish ovary (CCO) cells. Acyclovir was able to significantly inhibit the expression of viral genes related to CCV viral DNA synthesis and suppress viral replication at a safe concentration. Furthermore, acyclovir blocked the cytopathic effects and apoptosis induced by CCV, thereby maintaining the normal cellular morphological structure, as shown by the protection of CCO cells from the formation of apoptotic bodies or nuclear fragmentation. Moreover, reverse transcript quantitative polymerase chain reaction (RT-qPCR) demonstrated that acyclovir suppressed the expression of caspase 3, caspase 8 and caspase 9, while there was no significant impact on the expression of the apoptosis-inhibiting gene bcl-2 in CCV-infected cells. In addition, acyclovir did not promote the expression of immune-related genes such as MyD88, Mx1, IRF3, IRF7, IFN-I, NF-kB and IL-1ß, suggesting that the antiviral activity of acyclovir to CCV infection is not achieved by facilitating the expression of immune-related genes in CCO cells. Taken together, the results from this study suggest that acyclovir could effectively regulate CCV-induced infection, and thus is a promising therapeutic agent against CCV. Our results will aid our understanding of the pharmacological mechanisms of antiviral agents.


Assuntos
Aciclovir/administração & dosagem , Antivirais/administração & dosagem , Peixes-Gato/virologia , Doenças dos Peixes/tratamento farmacológico , Ictalurivirus/efeitos dos fármacos , Ovário/citologia , Replicação Viral/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Efeito Citopatogênico Viral/efeitos dos fármacos , Feminino , Doenças dos Peixes/imunologia , Doenças dos Peixes/fisiopatologia , Doenças dos Peixes/virologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Regulação Viral da Expressão Gênica/efeitos dos fármacos , Ictalurivirus/genética , Ictalurivirus/fisiologia , Ovário/virologia , Proteínas Virais/genética , Proteínas Virais/metabolismo
12.
Arch Virol ; 163(9): 2503-2506, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29802548

RESUMO

Three monoclonal antibodies (MAbs) (27E4, 17H2, 8B6) against channel catfish virus (CCV) were developed by immunizing Balb/C mice. Using indirect ELISA, these MAbs reacted only with CCV and not with three other fish viruses or nine fish cell lines. During western blotting analysis, MAb 27E4 recognized 170 kDa and 47 kDa proteins, while MAb 17H2 and MAb 8B6 recognized 47 kDa and 56 kDa proteins, respectively. Furthermore, a sandwich ELISA was developed for detection of CCV. The detection limit of the test was 105 TCID50/mL.


Assuntos
Anticorpos Monoclonais/química , Ensaio de Imunoadsorção Enzimática/métodos , Doenças dos Peixes/diagnóstico , Ictaluridae/virologia , Ictalurivirus/imunologia , Proteínas Virais/administração & dosagem , Animais , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/isolamento & purificação , Especificidade de Anticorpos , Linhagem Celular , Doenças dos Peixes/virologia , Ictalurivirus/isolamento & purificação , Imunização , Limite de Detecção , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Proteínas Virais/imunologia
13.
Arch Virol ; 163(4): 1083-1085, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29282547

RESUMO

Ictalurid herpesvirus 2 (IcHV-2) has been causing substantial losses in the black bullhead aquaculture industry since the 1990s. Using next-generation sequencing, the genome of IcHV-2 was completely sequenced and analysed in this study. The complete genome was found to be 142,925 bp in size, containing 77 predicted protein-coding regions, including 12 ORFs that appear to have a homologue in every alloherpesvirus genome sequenced to date. The genome organization of the IcHV-2 shows high similarity to that of IcHV-1, the founding member of the genus Ictalurivirus within the family Alloherpesviridae. A unique sequence region of 101 kbp is flanked by terminal direct repeats of 20 kbp. Thirteen of the 77 putative genes do not show homology to any known genes with sequences in public databases; six of them are found in the repeat regions. Analysis of the whole genome confirms the previously established taxonomic position of IcHV-2.


Assuntos
DNA Viral/genética , Doenças dos Peixes/virologia , Genoma Viral , Infecções por Herpesviridae/veterinária , Ictaluridae/virologia , Ictalurivirus/genética , Animais , Mapeamento Cromossômico , Tamanho do Genoma , Infecções por Herpesviridae/virologia , Ictalurivirus/classificação , Ictalurivirus/isolamento & purificação , Fases de Leitura Aberta , Filogenia , Sequências Repetidas Terminais , Sequenciamento Completo do Genoma
14.
Fish Shellfish Immunol ; 71: 58-68, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28970047

RESUMO

The channel catfish virus (CCV) can cause lethal hemorrhagic infection in juvenile channel catfish, thereby resulting in a huge economic loss to the fish industry. The genome of the CCV has been fully sequenced, and its prevalence is well documented. However, less is known about the molecular mechanisms and pathogenesis of the CCV. Herein, the channel catfish ovary cells (CCO) were infected with CCV and their transcriptomic sketches were analyzed using an RNA sequencing technique. In total, 72,686,438 clean reads were obtained from 73,231,128 sequence reads, which were further grouped into 747,168 contigs. These contigs were assembled into 49,119 unigenes, of which 20,912 and 18,333 unigenes were found in Nr and SwissProt databases and matched 15,911 and 14,625 distinctive proteins, respectively. From these, 3641 differentially expressed genes (DEGs), comprising 260 up-regulated and 3381 down-regulated genes, were found compared with the control (non-infected) cells. For verification, 16 DEGs were analyzed using qRT-PCR. The analysis of the DEGs and their related cellular signaling pathways revealed a substantial number of DEGs that were involved in the apoptosis pathway induced by CCV infection. The apoptosis pathways were further elucidated using standard apoptosis assays. The results showed that CCV could induce extrinsic apoptosis pathway (instead of a mitochondrial intrinsic apoptosis pathway) in CCO cells. This study helps our understanding of the pathogenesis of CCV and contributes to the prevention of CCV infection in channel catfish.


Assuntos
Proteínas de Peixes/genética , Expressão Gênica , Infecções por Herpesviridae/imunologia , Ictaluridae/genética , Ictaluridae/imunologia , Animais , Apoptose/imunologia , Células Cultivadas , Feminino , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Ictalurivirus/fisiologia , Reação em Cadeia da Polimerase em Tempo Real
15.
Dis Aquat Organ ; 124(2): 159-163, 2017 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-28425428

RESUMO

The channel catfish virus (CCV) is a pathogenic herpesvirus that infects channel catfish Ictalurus punctatus in pond aquaculture in the southeastern USA. Mannose-binding lectin (MBL), an innate immune protein, could play an important role in the innate response of channel catfish by binding to CCV. Cell cultures of CCV were grown in channel catfish ovary cells (CCOC). A dot-immunoblot enzyme-linked immunosorbent assay was done to determine the binding ability of 5 mo old channel catfish serum MBL (26.2 µg ml-1) to CCOC infected with CCV. Two separate nitrocellulose membrane blotting techniques were done using uninfected and infected CCOC. The uninfected CCOC decreased by 29.3 and 33.4% in their binding of channel catfish MBL when compared with infected CCOC using the 2 membrane procedures. The combined average binding ability of channel catfish MBL towards infected CCOC was therefore 31.4% greater when comparing the infected and uninfected CCOC. Normalization equation values of MBL for the 5 mo old catfish were compared for the 2 membrane binding procedures. The 2 normalization values were very close (142 and 150) in binding ability of MBL to the infected CCOC. The 5 mo catfish serum had twice the concentration of MBL (26.2 µg ml-1) compared to 7 mo catfish serum (13.2 µg ml-1), and the binding percentage of 5 mo serum was 2.4 times greater in infected than in uninfected cells. This demonstrates that the binding of channel catfish serum MBL to CCV is concentration dependent and is related to serum concentrations of MBL.


Assuntos
Infecções por Herpesviridae/veterinária , Ictaluridae/sangue , Ictalurivirus/imunologia , Imunidade Inata/fisiologia , Lectina de Ligação a Manose/fisiologia , Animais , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Feminino , Infecções por Herpesviridae/virologia , Immunoblotting , Lectina de Ligação a Manose/sangue , Ovário/citologia , Ligação Proteica
16.
J Fish Dis ; 40(10): 1363-1372, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28239935

RESUMO

Siberian sturgeon herpesvirus (SbSHV) was isolated in Russia for the first time in 2006. Nine SbSHV isolates were recovered from different fish hatcheries producing the same cytopathic effect in cell cultures, the same clinical signs and mortality kinetics in virus-infected fish and the same virus neutralization pattern and shared identical nucleotide sequences. In 2011, a new isolate was recovered from juvenile sturgeon, which caused completely different cytopathic effect. That isolate was not readily neutralized by Siberian sturgeon hyperimmune antisera, and its DNA was not recognized by the routine PCR developed for SbSHV detection. Molecular study of the novel isolate revealed that it was more closely related to North American Acipenserid herpesvirus 2 (AciHV-2) isolates from white sturgeon, while the genome sequences of the former SbSHV isolates showed high similarity to the AciHV-2 isolated from shortnose sturgeon. While clinical signs and mortality caused by the novel isolate in infected Siberian sturgeon were similar to those of the formerly described SbSHV isolates, the incubation period and mean time to death produced by the novel isolate were twice as long. The differences between the former isolates and the recent one suggest that a novel SbSHV strain emerged in Europe and the molecular findings imply its North American origin.


Assuntos
Doenças dos Peixes/virologia , Infecções por Herpesviridae/veterinária , Ictalurivirus/fisiologia , Sequência de Aminoácidos , Animais , Aquicultura , Proteínas do Capsídeo/genética , Peixes , Infecções por Herpesviridae/virologia , Ictalurivirus/genética , Federação Russa , Alinhamento de Sequência/veterinária
17.
J Immunol ; 196(6): 2677-89, 2016 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-26856701

RESUMO

Channel catfish, Ictalurus punctatus, leukocyte immune type receptors (LITRs) represent a multigene family that encodes Ig superfamily proteins that mediate activating or inhibitory signaling. In this study, we demonstrate the use of mAb CC41 to monitor viral cytotoxic responses in catfish and determine that CC41 binds to a subset of LITRs on the surface of catfish clonal CTLs. Homozygous gynogenetic catfish were immunized with channel catfish virus (CCV)-infected MHC-matched clonal T cells (G14D-CCV), and PBL were collected at various times after immunization for flow cytometric analyses. The percentage of CC41(+) cells was significantly increased 5 d after primary immunization with G14D-CCV and at 3 d after a booster immunization as compared with control fish only injected with G14D. Moreover, CC41(+) cells magnetically isolated from the PBL specifically killed CCV-infected targets as measured by (51)Cr release assays and expressed messages for CD3γδ, perforin, and at least one of the CD4-like receptors as analyzed by RNA flow cytometry. When MLC effector cells derived from a G14D-CCV-immunized fish were preincubated with CC41 mAb, killing of G14D-CCV targets was reduced by ∼40%, suggesting that at least some LITRs have a role in target cell recognition and/or cytotoxicity. The availability of a LITR-specific mAb has allowed, to our knowledge for the first time, functional characterization of LITRs in an autologous system. In addition, the identification of an LITR subset as a cytotoxic cell marker will allow for more effective monitoring of catfish immune responses to pathogens.


Assuntos
Doenças dos Peixes/imunologia , Infecções por Herpesviridae/imunologia , Ictaluridae , Ictalurivirus/imunologia , Leucócitos/imunologia , Receptores Imunológicos/metabolismo , Linfócitos T Citotóxicos/imunologia , Animais , Anticorpos Monoclonais/metabolismo , Proliferação de Células , Células Clonais , Citotoxicidade Imunológica , Imunização , Leucócitos/virologia , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Receptores Imunológicos/imunologia , Transdução de Sinais , Linfócitos T Citotóxicos/virologia
20.
Medisan ; 16(11)nov. 2012. tab
Artigo em Espanhol | CUMED | ID: cum-51919

RESUMO

El objetivo del presente trabajo fue evaluar niveles de proteínas totales y el factor de bioconcentración por exposición a metales en la Gambusia punctata. La especie fue muestreada en el ecosistema Filé y luego trasladada hacia condiciones de laboratorio, donde fueron diseñados 3 tratamientos a 2 réplicas con 25 ejemplares. Se determinó la concentración letal media (CL-50) como parámetro de toxicidad durante 48 horas de bioensayo. Los metales analizados fueron plomo y cadmio, cuantificados por espectroscopia de plasma inductivamente acoplados con vista axial. Transcurrido el experimento, la CL-50 correspondió a 0,1, ensayándose las concentraciones 0,06 y 5,78 mg/L, además del control negativo. Posteriormente se cuantificó el nivel de proteínas totales y los metales en agua, tejido y su relación mediante el factor de bioconcentración. El menor valor de proteínas fue ante la exposición al cadmio, con 43,9 por ciento de inhibición (p< 0,05) en comparación con el control; en el caso del plomo se determinó 2,5 por ciento de estimulación. Las mayores concentraciones en agua y tejido correspondieron a este último, no así para el factor de bioconcentración. Se concluyó que los resultados mostraron sensibilidad en la respuesta del contenido de proteínas totales y alta capacidad bioacumulativa para ambos metales(AU)


The aim of this study was to evaluate levels of total proteins and the bioconcentration factor by metal exposure in Gambusia punctata. The species was sampled in the ecosystem Filé and then transferred to the laboratory, where 3 treatments in 2 replications with 25 copies were designed. Mean lethal concentration (CL-50) was determined as a toxicity parameter for 48 hours of bioassay. The analyzed metals were lead and cadmium, quantified by plasma spectroscopy inductively coupled with axial view. After the experiment, the CL-50 corresponded to 0.1 and concentrations of 0.06 and 5.78 mg/L and the negative control were tested. Then the level of total proteins and metals in water, tissue and its relationship by means of the bioconcentration factor were quantified. The lower value of proteins was by exposure to cadmium with 43.9 percent of inhibition (p <0.05) compared with the control; for lead 2.5 percent of stimulation was determined. The highest concentrations in water and tissue corresponded to the latter, but not for the bioconcentration factor. It was concluded that the results showed sensitivity in the response of total protein content and a high bioaccumulative capacity for both metals


Assuntos
Animais , Metaloproteínas , Exposição Ambiental , Poecilia , Ciprinodontiformes , Ictalurivirus/patogenicidade , Poluição de Rios , Chumbo/análise , Cádmio/análise , Bioacumulação
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