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1.
PLoS One ; 16(3): e0247266, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33651823

RESUMO

Carnivore protoparvovirus-1 (CPPV-1), a viral species containing feline panleukopenia virus (FPV) and canine parvovirus (CPV) variants, are widely spread among domestic and wild carnivores causing systemic fatal diseases. Wild fishing cats (Prionailurus viverrinus), a globally vulnerable species, have been found dead. Postmortem examination of the carcasses revealed lesions in intestine, spleen and kidney. CPPV-1 antigen identification in these tissues, using polymerase chain reaction (PCR) and immunohistochemistry (IHC), supported the infection by the virus. PCR- and IHC-positivity in kidney tissues revealed atypical localization of the virus while in situ hybridization (ISH) and transmission electron microscopy (TEM) with the pop-off technique confirmed the first description of viral localization in kidneys. Complete genome characterization and deduced amino acid analysis of the obtained CPPV-1 from the fishing cats revealed FPV as a causative agent. The detected FPV sequences showed amino acid mutations at I566M and M569R in the capsid protein. Phylogenetic and evolutionary analyses of complete coding genome sequences revealed that the fishing cat CPPV-1 genomes are genetically clustered to the FPV genomes isolated from domestic cats in Thailand. Since the 1970s, these genomes have also been shown to share a genetic evolution with Chinese FPV strains. This study is the first evidence of CPPV-1 infection in fishing cats and it is the first to show its localization in the kidneys. These findings support the multi-host range of this parvovirus and suggest fatal CPPV-1 infections may result in other vulnerable wild carnivores.


Assuntos
Felidae/virologia , Vírus da Panleucopenia Felina/genética , Vírus da Panleucopenia Felina/patogenicidade , Animais , Animais Selvagens/virologia , Evolução Biológica , Proteínas do Capsídeo/genética , Carnívoros/genética , Gatos , Evolução Molecular , Panleucopenia Felina/virologia , Vírus da Panleucopenia Felina/isolamento & purificação , Especificidade de Hospedeiro , Rim/patologia , Rim/virologia , Mutação , Infecções por Parvoviridae/virologia , Parvovirus/genética , Parvovirus Canino/genética , Filogenia , Reação em Cadeia da Polimerase/veterinária , Tailândia
2.
J Vet Diagn Invest ; 32(6): 880-886, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32996420

RESUMO

Canine parvovirus 2 (CPV-2) and feline panleukopenia virus (FPLV) often cause acute enteric disease in their hosts. A simple, rapid, and effective method for the on-site detection of these viruses would be useful. We used a denaturation bubble-mediated strand exchange amplification (SEA) method to successfully detect CPV-2 and FPLV in fecal samples. SEA could detect as little as 3.6 pg/µL of CPV-2 and 6.6 pg/µL of FPLV genomic DNA following a 40-min incubation at an isothermal temperature of 61°C. Unlike PCR, SEA does not require complicated equipment, and positive samples produce a color change that can be visualized by the naked eye. Additionally, SEA is simpler than PCR because no extraction is needed, and heating of the fecal sample at 98°C can be performed with a heating block or water bath. This rapid and effective nucleic acid detection platform could be used as a point-of-care test for the detection of CPV-2 and FPLV.


Assuntos
Doenças do Cão/virologia , Fezes/virologia , Vírus da Panleucopenia Felina/isolamento & purificação , Panleucopenia Felina/diagnóstico , Infecções por Parvoviridae/veterinária , Parvovirus Canino/isolamento & purificação , Animais , Doenças do Gato/diagnóstico , Doenças do Gato/virologia , Gatos , Doenças do Cão/diagnóstico , Cães , Panleucopenia Felina/virologia , Vírus da Panleucopenia Felina/genética , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/virologia , Parvovirus Canino/genética , Reação em Cadeia da Polimerase/veterinária
3.
Biomed Res Int ; 2020: 9089768, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32923488

RESUMO

Purpose: We investigated sequences of the feline coronaviruses (FCoV), which include feline enteric coronavirus (FECV) and feline infectious peritonitis virus (FIPV), from China and other countries to gain insight into the adaptive evolution of this virus. Methods: Ascites samples from 31 cats with suspected FIP and feces samples from 8 healthy cats were screened for the presence of FCoV. Partial viral genome sequences, including parts of the nsp12-nsp14, S, N, and 7b genes, were obtained and aligned with additional sequences obtained from the GenBank database. Bayesian phylogenetic analysis was conducted, and the possibility of recombination within these sequences was assessed. Analysis of the levels of selection pressure experienced by these sequences was assessed using methods on both the PAML and Datamonkey platforms. Results: Of the 31 cats investigated, two suspected FIP cats and one healthy cat tested positive for FCoV. Phylogenetic analysis showed that all of the sequences from mainland China cluster together with a few sequences from the Netherlands as a distinct clade when analyzed with FCoV sequences from other countries. Fewer than 3 recombination breakpoints were detected in the nsp12-nsp14, S, N, and 7b genes, suggesting that analyses for positive selection could be conducted. A total of 4, 12, 4, and 4 positively selected sites were detected in the nsp12-nsp14, S, N, and 7b genes, respectively, with the previously described site 245 of the S gene, which distinguishes FIPV from FECV, being a positive selection site. Conversely, 106, 168, 25, and 17 negative selection sites in the nsp12-14, S, N, and 7b genes, respectively, were identified. Conclusion: Our study provides evidence that the FCoV genes encoding replicative, entry, and virulence proteins potentially experienced adaptive evolution. A greater number of sites in each gene experienced negative rather than positive selection, which suggests that most of the protein sequence must be conservatively maintained for virus survival. A few of the sites showing evidence of positive selection might be associated with the more severe pathology of FIPV or help these viruses survive other harmful conditions.


Assuntos
Adaptação Biológica/genética , Coronavirus Felino/genética , Peritonite Infecciosa Felina/virologia , Panleucopenia Felina/virologia , Genoma Viral/genética , Animais , Teorema de Bayes , Gatos , China , Coronavirus Felino/fisiologia , Evolução Molecular , Fezes/virologia , Peritonite Infecciosa Felina/patologia , Panleucopenia Felina/patologia , Mutação , Filogenia , Recombinação Genética , Seleção Genética , Proteínas Virais/genética
4.
J Vet Med Sci ; 82(9): 1243-1246, 2020 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-32759574

RESUMO

Feces obtained from 204 domestic cats with gastrointestinal symptoms were genetically examined for feline astrovirus (FeAstV) and feline parvovirus (FPV), both of which are known feline gastroenteric viruses. FeAstV detection rates were significantly higher in winter (44.4%) than in other seasons, and in cats under a year old (27.8%) than in a year or older ones (12.4%) (P<0.05). In contrast, no significant seasonal and age differences were obtained in FPV detection rates. Upon FeAstV ORF2 sequence analysis, the 23 present isolates were classified into the same clade (Mamastrovirus 2) as the 18 reference strains from other countries. Our findings suggest that FeAstV is already circulating in Japan, and it is more prevalent in juvenile cats in winter, unlike FPV.


Assuntos
Doenças do Gato/epidemiologia , Doenças do Cão/epidemiologia , Vírus da Panleucopenia Felina , Panleucopenia Felina/epidemiologia , Infecções por Parvoviridae/veterinária , Parvovirus Canino , Animais , Doenças do Gato/virologia , Gatos , Doenças do Cão/virologia , Cães , Panleucopenia Felina/virologia , Feminino , Japão/epidemiologia , Masculino , Infecções por Parvoviridae/epidemiologia , Prevalência
5.
Viruses ; 12(6)2020 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-32545689

RESUMO

Feline parvovirus (FPV) causes severe gastroenteritis and leukopenia in cats; the outcome is poor. Information regarding specific treatments is lacking. Class A CpG oligodeoxynucleotides (CpG-A) are short single-stranded DNAs, stimulating type I interferon production. In cats, CpG-A induced an antiviral response in vivo and inhibited FPV replication in vitro. The aim was to prospectively investigate the effects of CpG-A on survival, clinical score, hematological findings, antiviral response (cytokines), viremia, and fecal shedding (real-time qPCR) in cats naturally infected with FPV. Forty-two FPV-infected cats were randomized to receive 100 µg/kg of CpG-A (n = 22) or placebo (n = 20) subcutaneously, on admission and after 48 h. Blood and fecal samples were collected on admission, after 1, 3, and 7 days. All 22 cats showed short duration pain during CpG-A injections. The survival rate, clinical score, leukocyte and erythrocyte counts, viremia, and fecal shedding at any time-point did not differ between cats treated with CpG-A (50%) and placebo (40%). Antiviral myxovirus resistance (Mx) gene transcription increased in both groups from day 1 to 3 (p = 0.005). Antibodies against FPV on admission were associated with survival in cats (p = 0.002). In conclusion, CpG-A treatment did not improve the outcome in cats with FPV infection. FPV infection produced an antiviral response.


Assuntos
Vírus da Panleucopenia Felina/efeitos dos fármacos , Panleucopenia Felina/tratamento farmacológico , Oligodesoxirribonucleotídeos/administração & dosagem , Animais , Gatos , Contagem de Células , Panleucopenia Felina/sangue , Panleucopenia Felina/mortalidade , Panleucopenia Felina/virologia , Vírus da Panleucopenia Felina/fisiologia , Feminino , Leucócitos/citologia , Masculino , Estudos Prospectivos
6.
J Vet Sci ; 21(3): e43, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32476317

RESUMO

BACKGROUND: Canine parvovirus (CPV) and feline panleukopenia (FPV) cause severe intestinal disease and leukopenia. OBJECTIVES: In Korea, there have been a few studies on Korean FPV and CPV-2 strains. We attempted to investigate several genetic properties of FPV and CPV-2. METHODS: Several FPV and CPV sequences from around world were analyzed by Bayesian phylo-geographical analysis. RESULTS: The parvoviruses strains were newly classified into FPV, CPV 2-I, CPV 2-II, and CPV 2-III genotypes. In the strains isolated in this study, Gigucheon, Rara and Jun belong to the FPV, while Rachi strain belong to CPV 2-III. With respect to CPV type 2, the new genotypes are inconsistent with the previous genotype classifications (CPV-2a, -2b, and -2c). The root of CPV-I strains were inferred to be originated from a USA strain, while the CPV-II and III were derived from Italy strains that originated in the USA. Based on VP2 protein analysis, CPV 2-I included CPV-2a-like isolates only, as differentiated by the change in residue S297A/N. Almost CPV-2a isolates were classified into CPV 2-III, and a large portion of CPV-2c isolates was classified into CPV 2-II. Two residue substitutions F267Y and Y324I of the VP2 protein were characterized in the isolates of CPV 2-III only. CONCLUSIONS: We provided an updated insight on FPV and CPV-2 genotypes by molecular-based and our findings demonstrate the genetic characterization according to the new genotypes.


Assuntos
Doenças do Gato/virologia , Doenças do Cão/virologia , Vírus da Panleucopenia Felina/fisiologia , Genótipo , Infecções por Parvoviridae/veterinária , Animais , Gatos , Cães , Panleucopenia Felina/virologia , Vírus da Panleucopenia Felina/genética , Infecções por Parvoviridae/virologia , Parvovirus Canino/genética , Parvovirus Canino/fisiologia , Filogenia , República da Coreia
7.
Viruses ; 12(3)2020 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-32188011

RESUMO

Feline panleukopenia, caused by feline parvovirus (FPV), is a highly infectious disease characterized by leucopenia and hemorrhagic gastroenteritis that severely affects the health of large wild Felidae. In this study, tiger FPV virus-like particles (VLPs) were developed using the baculovirus expression system. The VP2 gene from an infected Siberian tiger (Panthera tigris altaica) was used as the target gene. The key amino acids of this gene were the same as those of FPV, whereas the 101st amino acid was the same as that of canine parvovirus. Indirect immunofluorescence assay (IFA) results demonstrated that the VP2 protein was successfully expressed. SDS-PAGE and Western blotting (WB) results showed that the target protein band was present at approximately 65 kDa. Electron micrograph analyses indicated that the tiger FPV VLPs were successfully assembled and were morphologically similar to natural parvovirus particles. The hemagglutination (HA) titer of the tiger FPV VLPs was as high as 1:218. The necropsy and tissue sections at the cat injection site suggested that the tiger FPV VLPs vaccine was safe. Antibody production was induced in cats after subcutaneous immunization, with a >1:210 hemagglutination inhibition (HI) titer that persisted for at least 12 months. These results demonstrate that tiger FPV VLPs might provide a vaccine to prevent FPV-associated disease in the tiger.


Assuntos
Vírus da Panleucopenia Felina/genética , Vírus da Panleucopenia Felina/imunologia , Panleucopenia Felina/imunologia , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/veterinária , Tigres/virologia , Animais , Proteínas do Capsídeo/genética , Gatos , Panleucopenia Felina/patologia , Panleucopenia Felina/virologia , Testes de Inibição da Hemaglutinação , Infecções por Parvoviridae/patologia , Infecções por Parvoviridae/virologia , Parvovirus Canino/genética , Parvovirus Canino/imunologia , Células Sf9 , Vacinas de Partículas Semelhantes a Vírus/imunologia
8.
Viruses ; 12(3)2020 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-32188115

RESUMO

Multiple, epizootic outbreaks of feline panleukopenia (FPL) caused by feline parvovirus(FPV) occurred in eastern Australia between 2014 and 2018. Most affected cats were unvaccinated.We hypothesised that low population immunity was a major driver of re-emergent FPL. The aim ofthis study was to (i) determine the prevalence and predictors of seroprotective titres to FPV amongshelter-housed and owned cats, and (ii) compare the prevalence of seroprotection between a regionaffected and unaffected by FPL outbreaks. FPV antibodies were detected by haemagglutinationinhibition assay on sera from 523 cats and titres ≥1:40 were considered protective. Socioeconomicindices based on postcode and census data were included in the risk factor analysis. The prevalenceof protective FPV antibody titres was high overall (94.3%), even though only 42% of cats wereknown to be vaccinated, and was not significantly different between outbreak and non-outbreakregions. On multivariable logistic regression analysis vaccinated cats were 29.94 times more likelyto have protective FPV titres than cats not known to be vaccinated. Cats from postcodes of relativelyless socioeconomic disadvantage were 5.93 times more likely to have protective FPV titres. Thepredictors identified for FPV seroprotective titres indicate targeted vaccination strategies in regionsof socioeconomic disadvantage would be beneficial to increase population immunity. The criticallevel of vaccine coverage required to halt FPV transmission and prevent FPL outbreaks should bedetermined.


Assuntos
Surtos de Doenças , Vírus da Panleucopenia Felina/imunologia , Panleucopenia Felina/epidemiologia , Panleucopenia Felina/imunologia , Animais , Anticorpos Antivirais/sangue , Austrália/epidemiologia , Gatos , Surtos de Doenças/prevenção & controle , Panleucopenia Felina/prevenção & controle , Panleucopenia Felina/virologia , Feminino , Masculino , Análise de Regressão , Fatores de Risco , Estudos Soroepidemiológicos , Vacinação/veterinária , Vacinas Virais
9.
PLoS One ; 15(1): e0227705, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31945103

RESUMO

Protoparvoviruses, widespread among cats and wild animals, are responsible for leukopenia. Feline panleukopenia virus (FPLV) in domestic cats is genetically diverse and some strains may differ from those used for vaccination. The presence of FPLV in two domestic cats from Hebei Province in China was identified by polymerase chain reaction. Samples from these animals were used to isolate FPLV strains in CRFK cells for genome sequencing. Phylogenetic analysis was performed to compare our isolates with available sequences of FPLV, mink parvovirus (MEV) and canine parvovirus (CPV). The isolated strains were closely related to strains of FPLV/MEV isolated in the 1960s. Our analysis also revealed that the evolutionary history of FPLV and MEV is characterized by local adaptations in the Vp2 gene. Thus, it is likely that new FPLV strains are emerging to evade the anti-FPLV immune response.


Assuntos
Antígenos Virais/imunologia , Gatos/virologia , Vírus da Panleucopenia Felina/genética , Panleucopenia Felina/virologia , Genes Virais/genética , Sequência de Aminoácidos/genética , Animais , Antígenos Virais/genética , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/imunologia , Linhagem Celular , China , Análise Mutacional de DNA , DNA Viral/genética , DNA Viral/isolamento & purificação , Cães/virologia , Evolução Molecular , Fezes/virologia , Panleucopenia Felina/imunologia , Vírus da Panleucopenia Felina/imunologia , Vírus da Panleucopenia Felina/patogenicidade , Vison/virologia , Mutação , Filogenia
10.
Viruses ; 11(12)2019 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-31847268

RESUMO

Feline panleukopenia (FPL), a frequently fatal disease of cats, is caused by feline parvovirus (FPV) or canine parvovirus (CPV). We investigated simultaneous outbreaks of FPL between 2014 and 2018 in Australia, New Zealand and the United Arab Emirates (UAE) where FPL outbreaks had not been reported for several decades. Case data from 989 cats and clinical samples from additional 113 cats were obtained to determine the cause of the outbreaks and epidemiological factors involved. Most cats with FPL were shelter-housed, 9 to 10 weeks old at diagnosis, unvaccinated, had not completed a primary vaccination series or had received vaccinations noncompliant with current guidelines. Analysis of parvoviral VP2 sequence data confirmed that all FPL cases were caused by FPV and not CPV. Phylogenetic analysis revealed that each of these outbreaks was caused by a distinct FPV, with two virus lineages present in eastern Australia and virus movement between different geographical locations. Viruses from the UAE outbreak formed a lineage of unknown origin. FPV vaccine virus was detected in the New Zealand cases, highlighting the difficulty of distinguishing the co-incidental shedding of vaccine virus in vaccinated cats. Inadequate vaccination coverage in shelter-housed cats was a common factor in all outbreaks, likely precipitating the multiple re-emergence of infection events.


Assuntos
Surtos de Doenças , Vírus da Panleucopenia Felina/classificação , Panleucopenia Felina/epidemiologia , Panleucopenia Felina/virologia , Animais , Austrália/epidemiologia , Gatos , DNA Viral , Geografia Médica , Nova Zelândia/epidemiologia , Estudos Retrospectivos , Análise de Sequência de DNA , Emirados Árabes Unidos/epidemiologia , Carga Viral
11.
J Virol Methods ; 271: 113679, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31216435

RESUMO

Feline panleukopenia caused by feline parvovirus (FPV), a single-stranded DNA virus, is typically highly contagious and often presents with lethal syndrome. The broad spectrum of possible hosts suggests its potential for transmission from animal to person through close contact with pets. FPV thus serves as an example of the importance of new rapid point-of-care field diagnostic tools for the control and prevention of transmission, especially among rare wild animals and pet cats. Recombinase polymerase amplification (RPA), as a real-time and isothermal method, could be a more affordable alternative to PCR when combined with a lateral flow dipstick (LFD) indicator. In this study, we report a novel FPV lateral flow dipstick RPA (LFD-RPA) instant detection method capable of detecting a range of different FPV strains. The LFD-RPA assay consists of specific primers, probe, and nucleic acid strip. It is capable of detecting 102 copies of target nucleic acid per reaction, which is one order of magnitude higher than the sensitivity of traditional PCR. The most suitable reaction conditions for this assay are at 38 ℃ for 15 min. This paper develops an efficient visual detection system that can eliminate the need for professional staff and expensive and sophisticated equipment for field detection.


Assuntos
Vírus da Panleucopenia Felina/isolamento & purificação , Panleucopenia Felina/diagnóstico , Técnicas de Diagnóstico Molecular/veterinária , Técnicas de Amplificação de Ácido Nucleico/veterinária , Recombinases/genética , Canal Anal/virologia , Animais , Gatos/virologia , Primers do DNA/genética , Sondas de DNA/genética , DNA Viral/isolamento & purificação , Panleucopenia Felina/virologia , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Sistemas Automatizados de Assistência Junto ao Leito , Fitas Reagentes , Sensibilidade e Especificidade
12.
Vet Clin North Am Small Anim Pract ; 49(4): 651-670, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30967253

RESUMO

Feline panleukopenia (FPL) is caused by a Carnivore protoparvovirus infection. Feline parvovirus (FPV) causes most cases. When Canine parvovirus 2 (CPV-2) first emerged, it could not replicate in cats. All current CPV variants (CPV-2a-c) can infect cats to cause subclinical disease or FPL. Feline panleukopenia has re-emerged in Australia in shelter cats associated with failure to vaccinate. Parvoviruses can remain latent in mononuclear cells post-infection. Molecular methods such as polymerase chain reaction are used to determine the infecting strain. Current perspectives on causes, epidemiology, diagnosis, treatment, prognostic indicators, and management of outbreaks in shelters are reviewed.


Assuntos
Doenças Transmissíveis Emergentes/veterinária , Panleucopenia Felina/epidemiologia , Animais , Gatos , Doenças Transmissíveis Emergentes/transmissão , Doenças Transmissíveis Emergentes/virologia , Panleucopenia Felina/patologia , Panleucopenia Felina/virologia , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/virologia , Parvovirus Canino
13.
Vet Pathol ; 56(4): 604-608, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30917745

RESUMO

Neurologic manifestations other than cerebellar hypoplasia are rarely associated with feline panleukopenia virus (FPV) infection in cats. Here the authors describe lymphoplasmacytic meningoencephalitis and neuronal necrosis in 2 cats autopsied after exhibiting ataxia and nystagmus. Gross changes consisted of cerebellar herniation through the foramen magnum, with flattening of cerebrocortical gyri and narrowing of sulci. Histologically, lymphoplasmacytic meningoencephalitis, extensive neuronal necrosis, and neuroaxonal degeneration with digestion chambers were present in the telencephalon and brain stem in both cats. Frozen brain tissue of both cats was positive for parvoviral antigen via fluorescent antibody testing, and formalin-fixed, paraffin-embedded tissue sections of brain were immunoreactive for parvovirus antigen and positive for parvoviral DNA on in situ hybridization. Frozen brain tissue from 1 case was positive for parvovirus NS1 and VP2 genes using conventional polymerase chain reaction, and subsequent DNA sequencing and phylogenetic analysis revealed that the viral strain was a FPV. Reverse transcription quantitative polymerase chain reaction on formalin-fixed, paraffin-embedded brain tissue revealed high levels of parvovirus in both cases, supporting an acute and active viral infection. Although rare, FPV infection should be considered in cases of lymphoplasmacytic meningoencephalitis and neuronal necrosis in cats.


Assuntos
Vírus da Panleucopenia Felina/isolamento & purificação , Panleucopenia Felina/patologia , Meningoencefalite/veterinária , Animais , Encéfalo/patologia , Gatos , Panleucopenia Felina/diagnóstico , Panleucopenia Felina/virologia , Vírus da Panleucopenia Felina/genética , Hibridização In Situ/veterinária , Meningoencefalite/diagnóstico , Meningoencefalite/virologia , Necrose/veterinária , Neurônios/patologia , Reação em Cadeia da Polimerase/veterinária
14.
Virus Genes ; 55(1): 95-103, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30519855

RESUMO

Feline panleukopenia virus (FPV) infects cats and can be fatal to kittens. There is evidence that canine parvovirus originated from FPV, which makes FPV important in studies of the family Parvoviridae. In the present study, the entire genome of FPV strain HH-1/86 was converted into a full-length infectious clone (pFPV). The FPV strain HH-1/86 has a 5123-nt single stranded DNA genome with a Y-shaped inverted 3' terminal repeat (ITR) and a U-shaped inverted 5' ITR. Feline kidney cells (F81) were transfected with the pFPV clone which contained a genetic marker, and a rescued virus was obtained (rFPV). The rFPV was identified by its cytopathic effects, indirect immunofluorescence, growth curve analysis, western blot assay and hemagglutination, and was indistinguishable from the parent virus. The FPV infectious clone will facilitate the study of pathogenicity and viral replication of FPV and the inter-species transmission of parvoviruses.


Assuntos
Vírus da Panleucopenia Felina/genética , Panleucopenia Felina/virologia , Genética Reversa , Animais , Gatos , Clonagem Molecular , DNA Viral , Marcadores Genéticos , Genoma Viral , Genômica/métodos , Hemaglutinação , Hemaglutininas Virais/metabolismo , Genética Reversa/métodos , Sequenciamento Completo do Genoma
15.
J Vet Med Sci ; 81(2): 252-255, 2019 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-30541981

RESUMO

To evaluate the accuracy of hemagglutination inhibition (HI) test as the index of feline panleukopenia virus (FPV)-protective ability, sera from 153 FPV-vaccinated cats aged ≥7 months with HI titer of <1:10-1:40, were examined for serum neutralizing (SN) antibody. SN antibody was detected (≥1:10) in 33 (62.3%) of 53 HI antibody-negative cats, and ranged <1:10-1:160. This suggests that FPV-antibody detection sensitivity of HI test is lower than SN test, and SN test is more suitable for the assessment of FPV-vaccine effect than HI test especially in cats with negative or low HI titer. SN titer was 1:32, FPV-protective threshold, or higher in all cats with HI titers of ≥1:20, suggesting it may be appropriate to set protective HI threshold at 1:20.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Vírus da Panleucopenia Felina/imunologia , Panleucopenia Felina/imunologia , Testes de Inibição da Hemaglutinação/veterinária , Animais , Gatos/imunologia , Gatos/virologia , Panleucopenia Felina/virologia , Vacinas Virais/imunologia
16.
Transbound Emerg Dis ; 65(6): 1429-1435, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30188020

RESUMO

Feline panleukopenia virus (FPV) and canine parvovirus (CPV) infections are highly contagious and cause serious enteric diseases, with high fatality rates of cats and dogs. Given the importance of cats as a potential source of genetic diversity for parvoviruses, parvovirus strains detected in cats with gastroenteritis signs were analysed, and molecular characterisation, sequence analysis and phylogeny were evaluated on the VP2 gene. The results showed that FPV, new CPV-2a, and CPV-2 are co-circulating in the cat population in Henan province of China. Moreover, CPV-2 strains (F2016020, and F2016021) with Ser297Ala substitution in VP2 protein was for the first time detected in cats with clinical gastroenteritis. This study provided new important findings about the evolutionary of parvovirus infection in domestic cats.


Assuntos
Animais Domésticos/virologia , Doenças do Gato/epidemiologia , Vírus da Panleucopenia Felina/genética , Panleucopenia Felina/epidemiologia , Animais , Doenças do Gato/virologia , Gatos , China/epidemiologia , Panleucopenia Felina/virologia , Vírus da Panleucopenia Felina/isolamento & purificação , Variação Genética , Filogenia , Proteínas Virais/genética
17.
Virol J ; 15(1): 45, 2018 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-29544546

RESUMO

BACKGROUND: The infection in dogs due to canine parvovirus (CPV), is a highly contagious one with high mortality rate. The present study was undertaken for a detailed genetic analysis of partial VP2 gene i.e., 630 bp isolated from rectal swab samples of infected domestic and stray dogs from all areas of district Faisalabad. Monitoring of viruses is important, as continuous prevalence of viral infection might be associated with emergence of new virulent strains. METHODS: In the present study, 40 rectal swab samples were collected from diarrheic dogs from different areas of district Faisalabad, Pakistan, in 2014-15 and screened for the presence of CPV by immunochromatography. Most of these dogs were stray dogs showing symptoms of diarrhea. Viral DNA was isolated and partial VP2 gene was amplified using gene specific primer pair Hfor/Hrev through PCR. Amplified fragments were cloned in pTZ57R/T (Fermentas) and completely sequenced. Sequences were analyzed and assembled by the Lasergene DNA analysis package (v8; DNAStar Inc., Madison, WI, USA). RESULTS: The results with immunochromatography showed that 33/40 (82%) of dogs were positive for CPV. We were able to amplify a fragment of 630 bp from 25 samples. In 25 samples the sequences of CPV-2a were detected showing the amino acid substitution Ser297Ala and presence of amino acid (426-Asn) in partial VP2 protein. Interestingly the BLAST analysis showed the of feline panleukopenia virus (FPV) sequences in 3 samples which were already positive for new CPV-2a, with 99% sequence homology to other FPV sequences present in GenBank. CONCLUSIONS: Phylogenetic analysis showed clustering of partial CPV-VP-2 gene with viruses from China, India, Japan and Uruguay identifying a new variant, whereas the 3 FPV sequences showed immediate ancestral relationship with viruses from Portugal, South Africa and USA. Interesting observation was that CPV are clustering away from the commercial vaccine strains. In this work we provide a better understanding of CPV prevailing in Pakistan at molecular level. The detection of FPV could be a case of real co-infection or a case of dual presence, due to ingestion of contaminated food.


Assuntos
Proteínas do Capsídeo/genética , Doenças do Cão/diagnóstico , Doenças do Cão/virologia , Vírus da Panleucopenia Felina/genética , Panleucopenia Felina/diagnóstico , Panleucopenia Felina/virologia , Variação Genética , Infecções por Parvoviridae/veterinária , Parvovirus Canino/genética , Animais , Gatos , DNA Viral , Cães , Vírus da Panleucopenia Felina/classificação , Vírus da Panleucopenia Felina/isolamento & purificação , Paquistão , Parvovirus Canino/classificação , Parvovirus Canino/isolamento & purificação , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA
19.
Vet Pathol ; 54(4): 669-675, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28622497

RESUMO

Canine parvovirus-2 (CPV-2) is nearly indistinguishable from feline panleukopenia virus (FPV) and is a well-known cause of viral myocarditis in young puppies; however, it is not known whether either FPV or CPV-2 naturally infects feline cardiomyocytes and causes myocarditis. Endomyocarditis (EMC) and left ventricular endomyocardial fibrosis (LVEF), clinically known as "endomyocardial restrictive cardiomyopathy," are important feline heart diseases suspected to have an infectious etiology. A continuum is suggested with EMC representing the acute reaction to an unknown infectious agent and LVEF the chronic manifestation of repair. The purpose of this study was to determine (1) whether there is natural parvovirus infection of the feline myocardium and (2) whether parvoviral infection is associated with feline EMC and/or LVEF. In a retrospective study, polymerase chain reaction and sequencing for the parvovirus VP1/2 gene was performed on archived heart tissue from cats with endomyocardial disease and controls. Similar methods were used prospectively on myocardial tissues from shelter-source kittens. Although 8 of 36 (22%; 95% confidence interval [CI], 11%-40%) shelter kittens had parvoviral DNA in myocardial tissue, VP1/2 DNA was not detected in 33 adult cases or 34 controls (95% CI, 0% to ∼11%). These findings were confirmed by in situ hybridization: adult cats did not have detectable parvovirus DNA, although rare intranuclear signal was confirmed in 7 of 8 shelter-source kittens. In kittens, parvovirus was not significantly associated with myocarditis, and in situ hybridization signal did not colocalize with inflammation. Although infection of cardiomyocytes was demonstrated in kittens, these data do not support a role for parvovirus in EMC-LVEF.


Assuntos
Cardiomiopatia Restritiva/veterinária , Vírus da Panleucopenia Felina , Panleucopenia Felina/patologia , Miocardite/veterinária , Animais , Cardiomiopatia Restritiva/patologia , Cardiomiopatia Restritiva/virologia , Gatos , Panleucopenia Felina/virologia , Feminino , Coração/virologia , Masculino , Miocardite/patologia , Miocardite/virologia , Miocárdio/patologia , Inclusão em Parafina/veterinária , Reação em Cadeia da Polimerase/veterinária , Estudos Retrospectivos
20.
Vet Microbiol ; 205: 92-98, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28622870

RESUMO

Six feline panleukopenia viruses (FPV) were detected in the intestinal samples from the 176 mink collected in China during 2015 to 2016, named MEV-SD1, MEV-SD2, MEV-SD3, MEV-SD4, MEV-SD5 and MEV-SD6. The VP2 genes of the isolates shared 98.9%-100% identity with the reference sequences. The substitution of residue V300A in VP2 protein differentiates the isolates from the reference MEVs, and A300 is a characteristic of FPV. Furthermore, phylogenetic analysis of VP2 genes indicated that the six isolates were clustered into the same branch of all the reference FPVs. The NS1 genes of the isolates shared 98.2%-100% identity with the reference sequences. The NS1 genes of the six isolates and the three reference FPVs formed one unique evolutionary branch. To clarify the pathogenicity of the isolates, animal experiments were performed on healthy mink, using MEV-SD1. As a result, the morbidity of the inoculated animals was 100% and the mortality was as high as 38.9%. It was implied that the FPV infection caused a high morbidity and mortality in mink and the inoculation dose had an effect on pathogenicity of MEV-SD1 in mink.


Assuntos
Vírus da Panleucopenia Felina/classificação , Panleucopenia Felina/virologia , Animais , Gatos , China , Vírus da Panleucopenia Felina/genética , Vírus da Panleucopenia Felina/isolamento & purificação , Vírus da Panleucopenia Felina/patogenicidade , Vison , Filogenia
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