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1.
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
2.
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
3.
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
4.
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
5.
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
6.
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
7.
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
8.
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
9.
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
10.
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
13.
Medisan ; 16(11): 1731-1735, nov. 2012.
Artigo em Espanhol | LILACS | ID: lil-660125

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.


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 , Bioacumulação , Ciprinodontiformes , Cádmio/análise , Exposição Ambiental , Ictalurivirus/patogenicidade , Metaloproteínas , Poecilia , Chumbo/análise , Poluição de Rios
14.
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
15.
Can J Microbiol ; 58(3): 271-7, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22356443

RESUMO

Channel catfish virus (CCV) is a viral pathogen of fry and fingerling channel catfish and can cause significant commercial loss. Previous studies have shown that the CCV virion contains at least 25 predicted structural proteins, including viral protein 10, which is encoded by the orf10 gene of the CCV. In this paper, the orf10 gene was expressed in Escherichia coli and used to produce a specific antibody. Western blot analysis confirmed that open reading frame 10 is an envelope protein. A viral neutralization assay demonstrated that open reading frame 10 antiserum was able to inhibit CCV infection of channel catfish ovary cells, suggesting that viral protein 10 is likely to play an important role in the CCV infection of channel catfish ovary cells.


Assuntos
Produtos do Gene env/genética , Ictalurivirus/genética , Animais , Linhagem Celular , Escherichia coli/genética , Produtos do Gene env/imunologia , Produtos do Gene env/metabolismo , Ictalurivirus/imunologia , Ictalurivirus/metabolismo , Soros Imunes/imunologia , Microscopia Imunoeletrônica , Testes de Neutralização , Fases de Leitura Aberta/genética , Vírion/metabolismo
16.
Hybridoma (Larchmt) ; 30(6): 555-8, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22149282

RESUMO

Three monoclonal antibodies (MAbs) against channel catfish virus (CCV) were generated from mice immunized with purified CCV. Western blot analysis revealed that the MAb 3G12 reacted with three CCV proteins of 94 kDa, 130 kDa, and 170 kDa; the MAb 4C4 reacted with two CCV proteins of 130 kDa and 170 kDa; and the MAb 4D4 reacted with two CCV proteins of 94 kDa and 98 kDa. Indirect immunofluorescence assay showed intense fluorescence in the CCV-infected channel catfish ovary (CCO) cells in areas corresponding to the location of granular structures. In addition, the three MAbs could completely neutralize CCV at a dilution of 1:500. This study demonstrated that these MAbs could recognize CCV specifically and will be useful in the development of diagnostic methods for the detection of fish CCV infection.


Assuntos
Anticorpos Monoclonais Murinos/imunologia , Especificidade de Anticorpos , Infecções por Herpesviridae/veterinária , Ictaluridae/virologia , Ictalurivirus/imunologia , Animais , Anticorpos Monoclonais Murinos/genética , Anticorpos Neutralizantes , Reações Antígeno-Anticorpo , Western Blotting , Linhagem Celular , Feminino , Fluorescência , Técnica Indireta de Fluorescência para Anticorpo , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/virologia , Hibridomas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Testes de Neutralização , Ovário/imunologia , Ovário/virologia
17.
Dis Aquat Organ ; 95(3): 189-201, 2011 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-21932530

RESUMO

Channel catfish virus (CCV, Ictalurid herpesvirus 1) and CCV disease have been extensively studied. Yet, little is known about CCV-host interaction after resolution of the primary infection. In order to determine potential recrudescence of CCV from latency, we established latency by exposing channel catfish juveniles with CCV or a thymidine kinase-negative recombinant (CCVlacZ) at a dose that caused less than 20% mortality. Then, we evaluated antibody response by serially sampling the same fish at 0 (pre-infection), 30, 60 and 90 d post challenge (DPC). We then attempted to induce viral recrudescence by intramuscular administration of dexamethasone and sampled the fish at 2, 4, 7, or 10 d post treatment. Recrudescence was evaluated by leukocyte co-cultivation and cell culture of tissue homogenates but no virus was detected. Western blot data demonstrated the highest number of seropositive fish by 30 DPC and a secondary antibody induction after dexamethasone treatment. The antigen specificity of the secondary response corresponded to viral proteins with molecular masses similar to those recognized by the same fish by 30 DPC. The recognized proteins were predominantly large, ranging from approximately 90 to >200 kDa. Expression analysis of selected virus genes at 90 DPC and following dexamethasone treatment demonstrated occasional immediate-early virus gene expression in peripheral blood leukocytes. Early and late gene expression was rarely detected. The combined data suggest restricted re-activation of CCV in our experimental system. Primary and secondary responses and virus gene expression were demonstrated in CCVlacZ-exposed fish but were less frequent than in CCV-exposed fish.


Assuntos
Anticorpos Antivirais/sangue , Dexametasona/toxicidade , Doenças dos Peixes/imunologia , Infecções por Herpesviridae/veterinária , Ictaluridae , Ictalurivirus , Animais , Doenças dos Peixes/virologia , Regulação Viral da Expressão Gênica , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/virologia , Interações Hospedeiro-Patógeno , Temperatura Alta/efeitos adversos , Imunossupressores/toxicidade , Recidiva , Latência Viral
18.
Arch Virol ; 156(12): 2291-6, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21935624

RESUMO

Sequencing of approximately one half of the genome of acipenserid herpesvirus 2 (AciHV-2), which is a member of the genus Ictalurivirus in the family Alloherpesviridae, revealed that the gene organization is very similar to that of ictalurid herpesvirus 1 (IcHV-1), the founder member of the genus. The sequenced region encodes the AciHV-2 homologues of IcHV-1 ORF24 to ORF69. It contains 46 predicted protein-coding regions, including 12 that seem to have a homologue in every alloherpesvirus genome sequenced to date. Phylogenetic tree reconstruction, based on the concatenated sequence of these conserved genes, implied that the family Alloherpesviridae is composed of three major clades and could be subdivided into three subfamilies.


Assuntos
Genoma Viral , Herpesviridae/classificação , Herpesviridae/genética , Ictalurivirus/classificação , Ictalurivirus/genética , Anfíbios/virologia , Animais , Sequência de Bases , Classificação , Primers do DNA/genética , DNA Viral/genética , Peixes/virologia , Dados de Sequência Molecular , Filogenia , Especificidade da Espécie
19.
Intervirology ; 54(5): 282-9, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21228534

RESUMO

OBJECTIVE: Partial genome sequences were determined and subjected to comparative analyses from two fish herpesviruses (HVs). Acipenserid (Aci) HV-2, originating from the white sturgeon (Acipenser transmontanus), and ictalurid (Ic) HV-2, isolated from the black bullhead (Ameiurus melas), are recently approved species of the genus Ictalurivirus of the family Alloherpesviridae. METHODS: An almost 8,000-base-pair fragment, spanning between the genes of the DNA polymerase and the ATPase subunit of the terminase, was sequenced from each virus. RESULTS: The size, position and orientation of 2 partial and 3 full open reading frames, contained in the studied genome fragment, proved to be similar to their counterparts in IcHV-1, the type species of the genus Ictalurivirus. Thus, a well-conserved genus-specific gene block was identified. In the members of two other genera (Cyprinivirus and Batrachovirus) of the family Alloherpesviridae, no such gene block could be found; the location and orientation of the homologous genes showed significant divergence. CONCLUSION: The results of phylogenetic calculations were in good agreement with the genome arrangements inasmuch as AciHV-2, IcHV-1 and -2 are monophyletic and separated from the lineages of the other two genera. The new sequence enabled the inclusion of a hitherto unassigned HV, that of the Australian pilchard, into a phylogenetic calculation.


Assuntos
Sequência Conservada , Genes Virais , Genoma Viral , Ictalurivirus/genética , Animais , Análise por Conglomerados , DNA Viral/química , DNA Viral/genética , DNA Polimerase Dirigida por DNA/genética , Endodesoxirribonucleases/genética , Peixes/virologia , Ictalurivirus/isolamento & purificação , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , Homologia de Sequência , Proteínas Virais/genética
20.
Virol J ; 7: 182, 2010 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-20691099

RESUMO

Viral-induced infectious diseases represent a major health threat and their control remains an unachieved goal, due in part to the limited availability of effective anti-viral drugs and measures. The use of natural products in drug manufacturing is an ancient and well-established practice. Marine organisms are known producers of pharmacological and anti-viral agents. In this study, a total of 20 extracts from marine microorganisms were evaluated for their antiviral activity. These extracts were tested against two mammalian viruses, herpes simplex virus (HSV-1) and vesicular stomatitis virus (VSV), using Vero cells as the cell culture system, and two marine virus counterparts, channel catfish virus (CCV) and snakehead rhabdovirus (SHRV), in their respective cell cultures (CCO and EPC). Evaluation of these extracts demonstrated that some possess antiviral potential. In sum, extracts 162M(4), 258M(1), 298M(4), 313(2), 331M(2), 367M(1) and 397(1) appear to be effective broad-spectrum antivirals with potential uses as prophylactic agents to prevent infection, as evident by their highly inhibitive effects against both virus types. Extract 313(2) shows the most potential in that it showed significantly high inhibition across all tested viruses. The samples tested in this study were crude extracts; therefore the development of antiviral application of the few potential extracts is dependent on future studies focused on the isolation of the active elements contained in these extracts.


Assuntos
Antivirais/farmacologia , Bactérias/química , Diatomáceas/química , Água do Mar/microbiologia , Vírus/efeitos dos fármacos , Animais , Antivirais/isolamento & purificação , Bactérias/isolamento & purificação , Linhagem Celular , Diatomáceas/isolamento & purificação , Herpesvirus Humano 1/efeitos dos fármacos , Humanos , Ictalurivirus/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Novirhabdovirus/efeitos dos fármacos , Vesiculovirus/efeitos dos fármacos
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