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1.
Virol J ; 18(1): 167, 2021 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-34391449

RESUMO

BACKGROUND: Poxviruses within the Capripoxvirus, Orthopoxvirus, and Parapoxvirus genera can infect livestock, with the two former having zoonotic importance. In addition, they induce similar clinical symptoms in common host species, creating a challenge for diagnosis. Although endemic in the country, poxvirus infections of small ruminants and cattle have received little attention in Botswana, with no prior use of molecular tools to diagnose and characterize the pathogens. METHODS: A high-resolution melting (HRM) assay was used to detect and differentiate poxviruses in skin biopsy and skin scab samples from four cattle, one sheep, and one goat. Molecular characterization of capripoxviruses and parapoxviruses was undertaken by sequence analysis of RPO30 and GPCR genes. RESULTS: The HRM assay revealed lumpy skin disease virus (LSDV) in three cattle samples, pseudocowpox virus (PCPV) in one cattle sample, and orf virus (ORFV) in one goat and one sheep sample. The phylogenetic analyses, based on the RPO30 and GPCR multiple sequence alignments showed that the LSDV sequences of Botswana were similar to common LSDV field isolates encountered in Africa, Asia, and Europe. The Botswana PCPV presented unique features and clustered between camel and cattle PCPV isolates. The Botswana ORFV sequence isolated from goat differed from the ORFV sequence isolated from sheep. CONCLUSIONS: This study is the first report on the genetic characterization of poxvirus diseases circulating in cattle, goats, and sheep in Botswana. It shows the importance of molecular methods to differentially diagnose poxvirus diseases of ruminants.


Assuntos
Vírus da Doença Nodular Cutânea/genética , Vírus do Orf , Infecções por Poxviridae , Poxviridae , Vírus da Pseudovaríola das Vacas , Animais , Botsuana/epidemiologia , Bovinos , Cabras , Vírus do Orf/genética , Filogenia , Poxviridae/genética , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/genética , Ruminantes , Ovinos , Doenças dos Ovinos/epidemiologia
2.
Arch Virol ; 166(1): 243-247, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33159590

RESUMO

Pseudocowpox is a zoonosis caused by pseudocowpox virus (PCPV), which mainly affects cows but can be an occupational disease of humans. The aim of the study was to validate a quantitative polymerase chain reaction (qPCR) assay for the detection of PCPV. The assay was able to detect up to 1000 copies of PCPV per µL in field samples, with a sensitivity of 80% and a specificity of 100%. We did not observe any cross-reactivity between PCPV-positive samples and samples that were positive for other genetically similar viruses. The repeatability and reproducibility were adequate according to parameters preestablished in official test validation manuals.


Assuntos
Reação em Cadeia da Polimerase/métodos , Infecções por Poxviridae/virologia , Vírus da Pseudovaríola das Vacas/genética , Animais , Bovinos , Doenças dos Bovinos/virologia , Humanos , Infecções por Poxviridae/veterinária , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Zoonoses/virologia
3.
Microb Pathog ; 143: 104122, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32169495

RESUMO

We studied the pathogenesis of Pseudocowpox virus (PCPV), a zoonotic parapoxvirus associated with mucocutaneous lesions in cattle. Inoculation of calves with PCPV isolate SD 76-65 intranasally (n = 6) or transdermally in the muzzle (n = 2) resulted in virus replication and shedding up to day 13 post-infection (pi). No local or systemic signs were observed in inoculated calves up to day 20pi, when the clinical monitoring was discontinued. However, from days 28-34 pi, seven (7/8) inoculated calves underwent an asynchronous clinical course characterized by development of a few (one or two) to countless papulo-pustular, erosive-fibrinous and scabby lesions in the muzzle, in some cases extending to the lips and gingiva. In some animals, the lesions coalesced, forming extensive fibrinotic/necrotic and scabby plaques covering almost entirely the muzzle. The clinical course lasted 8-15 days and spontaneously subsided after day 42pi. Infectious virus and/or viral DNA were detected in swabs collected from lesions of 5/8 animals between days 34 and 42pi. Histological examination of fragments collected from the muzzle lesions of two affected calves (day 36pi) revealed marked epidermal hyperplasia and severe orthokeratotic and parakeratotic hyperkeratosis, covered by thick scabs. The epidermis showed multifocal areas of keratinocyte coalescing necrosis and mild multifocal vacuolar degeneration. Sera of inoculated calves at 50pi showed partial virus neutralization at low dilutions, demonstrating seroconversion. The delayed and severe clinical course associated with virus persistence in lesions are novel findings and contribute for the understanding of PCPV pathogenesis.


Assuntos
Doenças dos Bovinos/virologia , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas , Animais , Bovinos , Doenças dos Bovinos/patologia , Face/patologia , Infecções por Poxviridae/patologia , Infecções por Poxviridae/virologia , Carga Viral/veterinária
4.
Virol J ; 17(1): 152, 2020 10 09.
Artigo em Inglês | MEDLINE | ID: mdl-33036619

RESUMO

BACKGROUND: Pseudocowpox virus (PCPV) of the genus Parapoxvirus in the family Poxviridae causes pseudocowpox in cattle worldwide and presents a zoonotic concern. Most poxviruses produce diseases of similar clinical signs in affected animals, which are impossible to differentiate clinically or by serology. It is, therefore, vital to use molecular assays to rapidly identify the causative agents of poxvirus infections. This study aimed to detect, diagnose, and characterize the causative agent of pox-like skin lesions in a cattle herd in Zambia, initially suspected to be infected with Lumpy Skin Disease virus. METHODS: We used a High-Resolution Melting (HRM) analysis assay to detect the PCPV genome and sequenced the major envelope protein (B2L gene) for comparative sequence and phylogenetic analysis. RESULTS: Our field investigations showed cattle presenting atypical skin lesions and high morbidity within the herd. The laboratory diagnosis, based on the HRM assay revealed PCPV DNA in the samples. Phylogenetic and comparative sequence analyses confirmed PCPV in the samples and revealed genomic differences between samples collected in 2017 and 2018 from the same farm. CONCLUSION: Our work is the first documented report of PCPV in Zambia. It shows the strength of molecular methods to diagnose pox-like infections in cattle and discriminate between diseases causing similar clinical signs. This rapid and accurate diagnosis improves the response time for more accurate veterinary interventions.


Assuntos
Bovinos/virologia , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/genética , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Animais , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Filogenia , Infecções por Poxviridae/virologia , Vírus da Pseudovaríola das Vacas/classificação , Pele/patologia , Pele/virologia , Zâmbia/epidemiologia
5.
Arch Virol ; 165(11): 2659-2664, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32880730

RESUMO

Two cases of coinfection with bovine papular stomatitis virus (BPSV) and pseudocowpox virus (PCPV) in dairy calves in Tochigi Prefecture, Japan, are reported. Sequences of BPSV and PCPV were simultaneously detected in the same polymerase chain reaction (PCR) amplicons, which were obtained from the DNA of two dairy calves using a pan-parapoxvirus primer set. PCR amplification using BPSV- and PCPV-specific primer sets were able to distinguish between the two viruses in coinfected clinical samples. Based on these data, further studies on the occurrence BPSV/PCPV coinfections in cattle in Japan are warranted.


Assuntos
Doenças dos Bovinos/virologia , Parapoxvirus/isolamento & purificação , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Animais , Bovinos , Coinfecção/virologia , Feminino , Japão , Masculino , Parapoxvirus/genética , Reação em Cadeia da Polimerase/veterinária , Infecções por Poxviridae/virologia , Vírus da Pseudovaríola das Vacas/genética
6.
BMC Vet Res ; 16(1): 241, 2020 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-32660468

RESUMO

BACKGROUND: The present report describes a case of pseudocowpox virus (PCPV) infection in a seven-year-old female bison euthanized due to a history of declining condition and sores on the vulva and udder. CASE PRESENTATION: External examination revealed multifocal, raised, keratinized plaques (0.5-2 cm) covering the skin of the ventral surface of the tail, perineum, caudoventral abdomen, udder, both inguinal recesses, and the medial aspects of both thighs. No significant gross lesions were present in the reminder of the tissues examined. Histopathological examination of the affected skin showed moderate epidermal hyperplasia with rete pegs, marked parakeratotic hyperkeratosis with crusts of degenerate neutrophils and cell debris, and few epithelial cells undergoing ballooning degeneration with occasional eosinophilic intracytoplasmic inclusion bodies (3-5 µm Bollinger body). Negative staining electron microscopy from skin revealed typical Parapoxvirus (PPV) particles, which were also confirmed by real-time PCR (Ct =18.6). Metagenomic analysis of the skin samples revealed only poxviruses. The bison parapox B2L envelope gene clustered with other parapox sequences identified from ruminants. CONCLUSIONS: This is the first report of PCPV virus infection in an American bison. Identification of novel susceptible hosts of parapox viruses sheds light on the viral evolution and highlights the importance of potential economic impact of this disease to the bison industry.


Assuntos
Bison , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Animais , DNA Viral/análise , Feminino , Kansas , Microscopia Eletrônica , Infecções por Poxviridae/virologia , Vírus da Pseudovaríola das Vacas/genética , Vírus da Pseudovaríola das Vacas/ultraestrutura , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Dermatopatias Virais/patologia , Dermatopatias Virais/veterinária
8.
Virol J ; 14(1): 172, 2017 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-28874200

RESUMO

BACKGROUND: Pseudocowpox virus (PCPV) infects cattle worldwide with zoonotic potential but has not been isolated in Japan. Thus, the epidemiological status of PCPV infection in cattle is undetermined. RESULTS: In May 2016, a cattle in a farm in Yamaguchi Prefecture showed white vesicles and hyperemia in the mucosa under the tongue surface, but not on the teats and coronary cushions. A parapoxvirus was isolated from the oral lesion swab and was genetically characterized based on the full-length sequence of B2L gene encoding viral envelope. Phylogenetic analysis showed that the isolated virus was classified into PCPV. CONCLUSION: This case indicates its potential spread in Japan. This is the first report of isolation of PCPV in Japan.


Assuntos
Doenças dos Bovinos/virologia , Filogenia , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/classificação , Vírus da Pseudovaríola das Vacas/genética , Proteínas do Envelope Viral/genética , Animais , Bovinos , Doenças dos Bovinos/patologia , Linhagem Celular , Células Cultivadas , Cricetinae , Feminino , Japão , Reação em Cadeia da Polimerase , Infecções por Poxviridae/patologia , Infecções por Poxviridae/virologia , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Homologia de Sequência
9.
Cytokine ; 61(2): 356-9, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23306428

RESUMO

Cellular interleukin-10 (IL-10) gene from the peripheral blood mononuclear cells of the healthy Dromedary camel (Camelus dromedarius) and viral IL-10 (vIL-10) from the skin scabs of the Dromedary camels infected with contagious ecthyma (a parapoxviral infection in the camels) were amplified by polymerase chain reaction, cloned and characterized. Sequence analysis revealed that the open reading frame (ORF) of dromedarian camel IL-10 is 537 bp in length, encoding 178 amino acid polypeptide while open reading frame of vIL-10 from camel is 561 bp, encoding 187 amino acid polypeptide. The Dromedary camel IL-10 exhibited 62.6% and 68.5% sequence identity at the nucleotide and amino acid level, respectively, with vIL-10 from camel. Sequence analysis also revealed that the Dromedary camel IL-10 shared 99.4% and 98.3% identity at the nucleotide and amino acid level, respectively, with the Bactrian camel (Camelus bactrianus). But vIL-10 from camel shared 84.7% and 83.4% sequence identity at the nucleotide and amino acid level, respectively, with vIL-10 from reindeer (Rangifer tarandus), which is a ruminant species belonging to the order Artiodactyla. The present study was conducted to evaluate the evolutionary origin of the camel parapoxvirus with parapoxviruses of cattle and sheep and the resultant sequence analysis revealed that camel parapoxvirus is closely related to cattle parapoxvirus than sheep parapoxvirus (Orf virus).


Assuntos
Camelus/imunologia , Camelus/virologia , Interleucina-10/imunologia , Vírus da Pseudovaríola das Vacas/imunologia , Sequência de Aminoácidos , Animais , Interleucina-10/química , Masculino , Dados de Sequência Molecular , Infecções por Poxviridae/veterinária , Infecções por Poxviridae/virologia , Alinhamento de Sequência
10.
Braz J Microbiol ; 53(2): 1077-1080, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35220553

RESUMO

The pseudocowpox virus (PCPV) is recognized for causing exanthematic lesions in cattle and humans. The diagnosis is important because it is a zoonosis and its clinical signs can be confused with foot-and-mouth disease, a high-impact bovine disease in livestock. The objective of this work is to validate a SYBR Green qPCR and a conventional PCR for virus detection in bovine samples. Detection limit tests, repeatability, reproducibility, sensitivity, and specificity were compared. When two analysts were compared, results demonstrated that training and pipetting influence the repeatability. The qPCR was more sensitive than conventional PCR but showed nonspecific reactions distinguishable by the melting curve. Both showed high repeatability and reproducibility.


Assuntos
Doenças dos Bovinos , Infecções por Poxviridae , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Patologia Molecular , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/genética , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
11.
J Gen Virol ; 91(Pt 6): 1560-76, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20107016

RESUMO

Parapoxviruses (PPV), of the family Poxviridae, cause a pustular cutaneous disease in sheep and goats (orf virus, ORFV) and cattle (pseudocowpoxvirus, PCPV and bovine papular stomatitis virus, BPSV). Here, we present the first genomic sequence of a reference strain of PCPV (VR634) along with the genomic sequence of a PPV (F00.120R) isolated in Finland from reindeer (Rangifer tarandus tarandus). The F00.120R and VR634 genomes are 135 and 145 kb in length and contain 131 and 134 putative genes, respectively, with their genome organization being similar to that of other PPVs. The predicted proteins of F00.120R and VR634 have an average amino acid sequence identity of over 95%, whereas they share only 88 and 73% amino acid identity with the ORFV and BPSV proteomes, respectively. The most notable differences were found near the genome termini. F00.120R lacks six and VR634 lacks three genes seen near the right terminus of other PPVs. Four genes at the left end of F00.120R and one in the middle of both genomes appear to be fragmented paralogues of other genes within the genome. VR634 has larger than expected inverted terminal repeats possibly as a result of genomic rearrangements. The high G+C content (64%) of these two viruses along with amino acid sequence comparisons and whole genome phylogenetic analyses confirm the classification of PCPV as a separate species within the genus Parapoxvirus and verify that the virus responsible for an outbreak of contagious stomatitis in reindeer over the winter of 1999-2000 can be classified as PCPV.


Assuntos
DNA Viral/química , DNA Viral/genética , Genoma Viral , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/genética , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Rena/virologia , Sequência de Aminoácidos , Animais , Composição de Bases , Análise por Conglomerados , Finlândia , Ordem dos Genes , Genes Virais , Dados de Sequência Molecular , Filogenia , Infecções por Poxviridae/virologia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Sintenia , Sequências Repetidas Terminais , Proteínas Virais/genética
12.
Anal Chem ; 80(8): 2817-25, 2008 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-18303918

RESUMO

Surface-enhanced Raman spectroscopy (SERS) can be made an attractive approach for identification of Raman-active compounds and biological materials (i.e., toxins, viruses, or intact bacterial cells/spores) through development of reproducible, spatially uniform SERS-active substrates. Recently, reproducible (from substrate-to-substrate), spatially homogeneous (over large areas) SERS-active substrates have been commercialized and are now available in the marketplace. We have utilized these patterned surfaces to acquire SERS spectral signatures of intact bovine papular stomatitis, pseudocowpox, and Yaba monkey tumor viruses. Salient spectral signature features make it possible to discriminate among these genetically distinct Poxviridae-Chordopoxvirinae virions. In addition, partial least-squares, a multivariate calibration method, has been used to develop personal computer-borne algorithms useful for classification of unknown Parapoxvirus (e.g., bovine papular stomatitis virus and pseudocowpox virus) samples based solely on SERS spectral signatures. To our knowledge, this is the first report detailing application of these commercial-off-the-shelf (COTS) SERS-active substrates to identification of intact poxviruses.


Assuntos
Parapoxvirus/química , Análise Espectral Raman/métodos , Vírion/química , Animais , Bovinos , Cercopithecus , Parapoxvirus/isolamento & purificação , Vírus da Pseudovaríola das Vacas/química , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Vírion/isolamento & purificação , Vírus do Tumor do Macaco de Yaba/química , Vírus do Tumor do Macaco de Yaba/isolamento & purificação
13.
Yao Xue Xue Bao ; 43(3): 253-8, 2008 Mar.
Artigo em Zh | MEDLINE | ID: mdl-18630260

RESUMO

This study is to establish a cell-based pharmacological model targeting HIV-1 replication for compounds screening and to screen compounds randomly selected from compounds library by using this pseudotyped viral system. The cell-based HIV-1 replication pharmacological model was set up by HIV-1 core packed with vesicular stomatitis virus glycoprotein. The level of HIV-1 replication was presented by reporter genes expression (luciferase activity or percentage of GFP positive cells). When a compound has inhibitory effect on VSVG/HIV model, VSVG/MLV model would be used to test for specificity. Vesicular stomatitis virus glycoprotein can efficiently mediate HIV core into a wide range of host cells. Expression level of reporter genes showed dose-dependent manner with virion dilution. Among 500 compounds, three compounds dose-dependently inhibit HIV-1 replication, but not MLV replication. VSVG/HIV pseudotyped viral system can be used as a pharmacological model for HIV-1 replication inhibitor screening. Compounds 2-methylthio-5-(4-methylbenzo)amido-l,3,4-thiadiazole, N-(3-hydroxyphenyl)-2-(4-isobutylphenyl) propionamide, and N-(4-picolyl)-4-methylbenzenesulfonamide can specifically inhibit HIV-1 replication with IC50 of 1.92, 5.38, and 3.39 micromol L(-1) respectively.


Assuntos
Fármacos Anti-HIV/farmacologia , Replicação do DNA/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos/métodos , HIV-1/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Didanosina/farmacologia , Genes Reporter/efeitos dos fármacos , Genes Reporter/genética , HIV-1/fisiologia , Humanos , Lamivudina/farmacologia , Vírus da Pseudovaríola das Vacas , Células Tumorais Cultivadas , Zidovudina/farmacologia
14.
J Vet Diagn Invest ; 30(2): 256-259, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29153035

RESUMO

Species Pseudocowpox virus (PCPV; family Poxviridae) is known to cause pustular cutaneous disease in cattle. We describe an outbreak of pseudocowpox with an unusual clinical picture in a free-stall dairy herd of ~80 cows. Approximately 90% of the cows had vesicles, erosions, papules, and scabs on the vulva and vaginal mucosa. Histologic analysis of biopsy tissues indicated a primary, although not specified, viral infection. Transmission electron microscopy revealed parapoxvirus particles in both tissue and vesicular materials. Deep sequencing analysis of extracted DNA from swabbed vesicle areas gave a contig of nearly 120,000 nucleotides, matching the PCPV strain VR 634 with 100% identity. Analyses confirmed the absence of other potential causes of pustular vulvovaginitis such as bovine herpesvirus 1 and Ureaplasma diversum. A rolling cow brush was suspected to be the fomite.


Assuntos
Doenças dos Bovinos/epidemiologia , Surtos de Doenças/veterinária , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Vulvovaginite/veterinária , Animais , Bovinos , Doenças dos Bovinos/patologia , Doenças dos Bovinos/virologia , Indústria de Laticínios , Feminino , Infecções por Poxviridae/epidemiologia , Suécia/epidemiologia , Vulvovaginite/epidemiologia
15.
Vector Borne Zoonotic Dis ; 7(2): 257-60, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17627446

RESUMO

We have determined the nucleotide sequence of DNA extracted from pustules, saliva, and blood of camels presenting with contagious ecthyma, in Bahrain and also from a sample (SACamel) of infected tissue from a camel that had presented with contagious ecthyma in 1998 in Saudi Arabia (1). Sequence homologies and phylogenetic analysis showed that this extracted DNA was more closely related to Pseudocowpox virus (PCPV) than Orf virus (ORFV), which infects sheep, goats, and other animal species. The phylogeny also demonstrated that PCPV in Arabian camels was phylogenetically distinct from, and circulates independently of, ruminant-associated PCPV from Europe.


Assuntos
Camelus/virologia , Ectima Contagioso/epidemiologia , Filogenia , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/classificação , Animais , Barein/epidemiologia , Sequência de Bases , DNA Viral/química , Surtos de Doenças/veterinária , Ectima Contagioso/virologia , Feminino , Masculino , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/virologia , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Arábia Saudita/epidemiologia
16.
An Bras Dermatol ; 92(6): 838-840, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29364443

RESUMO

Milker's nodule is an occupational dermatovirose caused by Parapoxvirus, which is self-limited and, due to the lack of information of health professionals, may lead to underdiagnosis. We present two cases with exuberant manifestations and classic histopathologic findings. Case 1: Male, 19 years of age, milker, presented nodules and blisters on his palm for 15 days. Case 2: Male, 33 years of age, administrative assistant, presented erythematous nodules on his palms with lymphangitis for 5 days. He had milked a cow one week before the onset of the lesions. In both cases, the histopathology was representative and coincident with the clinical hypothesis. The lesions have presented complete involution. Milker's nodule diagnosis is based on clinical presentation, epidemiology, and histopathology. The knowledge of this disease is essential for its correct diagnosis, as well as to guide the implementation of public health measures and the appropriate treatment of sick cattle.


Assuntos
Dermatoses da Mão/patologia , Dermatoses da Mão/virologia , Infecções por Poxviridae/patologia , Adulto , Biópsia , Dermatite Ocupacional/patologia , Dermatite Ocupacional/virologia , Epiderme/patologia , Humanos , Masculino , Vírus da Pseudovaríola das Vacas/patogenicidade , Adulto Jovem
17.
J Virol Methods ; 239: 38-41, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27793645

RESUMO

Exanthematic and papulo-vesicular lesions in the udder and teats of milking cows are fairly common in some Brazilian dairies, especially those with poor sanitary conditions and hand milking. The orthopoxvirus Vaccinia virus (VACV) and the parapoxviruses Pseudocowpox virus (PCPV) and Bovine popular stomatitis virus (BPSV) have been frequently associated with such conditions. Elsewhere, Bovine herpesvirus 2 (BoHV-2) has also been associated with similar clinical signs. Thus, we herein describe a conventional multiplex PCR designed to detect the genome of these viruses in clinical samples while differentiating among them by amplicon size. For this, primer sets targeting the orthopoxvirus vascular growth factor (amplicon size 292bp), PCPV (374bp) and BSPV (607bp) B2L genes, and the BoHV-2 DNA polymerase gene (138bp) were selected. The chosen primers anneal within the same temperature range and do not interfere with each other during the PCR amplification. PCR conditions were initially standardized for each agent in individual PCR reactions firstly using the target virus as positive control followed by using a mixture of all four virues. Lastly, a multiplex PCR containing the four sets of primers was set up to amplify all four targeted viruses in one reaction. The multiplex PCR was able to detect DNA extracted from cell culture supernatants containing 20 TCID50 of BoHV-2 and 50 TCID50 of VACV. Further, the test could detect the viral genomes in 1:10, 1:50 and 1:1000 dilutions of total DNA extracted from clinical specimens (e.g. scabs, crusts) of natural cases (PCPV, VACV and BPSV) and 1:10 dilutions of DNA extracted from scabs collected from BoHV-2 experimentally infected cattle. A possible amplification of other orthopoxviruses, predicted by in silico analysis, was considered to not represent an important pitfall since these are exotic in Brazil, very rare, or viruses not associated with cattle. For definitive agent identification amplicon sequencing needs to be conducted. Thus, this multiplex PCR seems suitable for initial detection and identification of the agents involved in exanthematic and vesicular disease, providing a sensitive and specific diagnosis for such conditions in dairy cows.


Assuntos
Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/virologia , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções por Poxviridae/veterinária , Animais , Brasil , Bovinos , Primers do DNA , DNA Viral/genética , Genes Virais , Genoma Viral , Orthopoxvirus/genética , Orthopoxvirus/isolamento & purificação , Parapoxvirus/genética , Parapoxvirus/isolamento & purificação , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/virologia , Vírus da Pseudovaríola das Vacas/genética , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Vaccinia virus/genética , Vaccinia virus/isolamento & purificação
18.
Vet Q ; 37(1): 16-22, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27774853

RESUMO

BACKGROUND: Parapoxviruses are zoonotic viruses that infect cattle, goats and sheep; there have also been reports of infections in camels, domestic cats and seals. OBJECTIVE: The objective of this report was to describe a case of vesicular disease caused by pseudocowpox virus (PCPV) in water buffalo (Bubalus bubalis) in Brazil. ANIMALS: Sixty buffalo less than 6 months old exhibited ulcers and widespread peeling of the tongue epithelium. There were no cases of vesicular disease in pigs or horses on the same property. METHODS: Samples were analysed by PCR and sequencing. Phylogenetic analysis in MEGA 7.01 was reconstructed using major envelope protein (B2L) by the Tamura three-parameter nucleotide substitution model and the maximum likelihood and neighbor joining models, both with 1000 bootstrap replicates. The genetic distance between the groups was analysed in MEGA using the maximum composite likelihood model. The rate variation among sites was modeled using gamma distribution. RESULTS: The presence of PCPV in the buffalo herd could be demonstrated in epithelium and serum. The minimum genetic distance between the isolated PCPV strain (262-2016) and orf virus and bovine papular stomatitis virus was 6.7% and 18.4%, respectively. The maximum genetic distance calculated was 4.6% when compared with a PCPV detected in a camel. Conclusions/Clinical Importance: The peculiar position of the isolated strain in the phylogenetic trees does not necessarily indicate a different kind of PCPV that infects buffalo. More samples from cattle and buffalo in Brazil must be sequenced and compared to verify if PCPV from buffalo are genetically different from samples derived from cattle.


Assuntos
Búfalos , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas/fisiologia , Animais , Brasil/epidemiologia , Incidência , Filogenia , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/virologia , Análise de Sequência de DNA/veterinária , Proteínas do Envelope Viral/genética
19.
Pesqui. vet. bras ; 42: e07014, 2022. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1360625

RESUMO

A retrospective study of poxvirus infections diagnosed in cattle from Goiás state (GO), Brazil, from 2010 to 2018, was performed. All cases have been investigated by the GO Official Veterinary Service (Agrodefesa), from which technical forms and protocols of veterinary diagnosis laboratories were reviewed. In most cases, samples of oral or cutaneous tissues and/or swabs were submitted for virological diagnosis by polymerase chain reaction (PCR) and/or virus isolation. Thirty seven outbreaks/cases of vesicular disease were notified in cattle of 25 counties; in 33 cases the animals presented lesions clinically compatible with poxviruses. The etiology of 25 out of 33 outbreaks/cases was confirmed as poxviruses by PCR and/or viral isolation: 13 as bovine vaccinia virus (VACV), six as pseudocowpox virus (PCPV), five as bovine papular stomatitis virus (BPSV) and one coinfection (VACV and an Orf virus-like parapoxvirus). The laboratory confirmed that cases occurred mainly in dairy cattle (19/25) and during the dry season (22/25). In adult cattle, gross changes were observed mainly in the teats and udder and included vesicles, ulcers, crusts, papules and scars and varied of type, severity and affected region, depending on the poxvirus species. In calves, the main lesions were ulcers in the mouth and muzzle. Zoonotic lesions compatible with poxvirus infections were observed for all diagnosed poxviruses, affecting especially the hands of milkers and other farm workers. Our data demonstrate the sanitary and economic relevance of these diseases and the wide circulation of different poxviruses in cattle from GO.(AU)


Foi realizado um estudo retrospectivo das infecções por poxvírus diagnosticadas em bovinos do estado de Goiás (GO), entre 2010 e 2018. Todos os casos foram investigados pela Agência Goiana de Defesa Agropecuária (Agrodefesa). Foram revisados formulários técnicos e protocolos de laboratórios de diagnóstico veterinário. Na maioria dos casos, amostras de tecidos orais ou cutâneos e/ou swabs foram encaminhadas para diagnóstico virológico. Foram notificados 37 surtos/casos de doença vesicular em bovinos em 25 municípios; em 33 casos os animais apresentavam lesões clinicamente compatíveis com poxvírus. A etiologia de 25 de 33 surtos/casos foi confirmada como poxvírus por PCR e/ou isolamento viral: 13 como vírus vaccínia (VACV), seis como vírus pseudocowpox (PCPV), cinco como vírus da estomatite papular bovina (BPSV) e um caso de coinfecção (VACV e um parapoxvírus semelhante ao Orf vírus). Os casos confirmados laboratorialmente ocorreram principalmente em bovinos leiteiros (19/25) e durante a estação seca (22/25). Em bovinos adultos, alterações macroscópicas foram observadas principalmente nas tetas e úbere e incluíram vesículas, úlceras, crostas, pápulas e cicatrizes e variaram quanto ao tipo, gravidade e região afetada, dependendo da espécie do poxvírus. Em bezerros, as principais lesões foram úlceras na boca e focinho. Lesões zoonóticas compatíveis com infecção por poxvírus foram observadas em todas as poxviroses diagnosticadas, afetando principalmente as mãos dos ordenhadores e outros trabalhadores rurais. Nossos dados demonstram a relevância sanitária e econômica dessas doenças e a ampla circulação de diferentes poxvírus em bovinos de GO.(AU)


Assuntos
Humanos , Animais , Bovinos , Vaccinia virus/isolamento & purificação , Parapoxvirus/isolamento & purificação , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/patologia , Infecções por Poxviridae/epidemiologia , Coinfecção/veterinária , Zoonoses Virais
20.
Pesqui. vet. bras ; 42: e07014, 2022. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1487701

RESUMO

A retrospective study of poxvirus infections diagnosed in cattle from Goiás state (GO), Brazil, from 2010 to 2018, was performed. All cases have been investigated by the GO Official Veterinary Service (Agrodefesa), from which technical forms and protocols of veterinary diagnosis laboratories were reviewed. In most cases, samples of oral or cutaneous tissues and/or swabs were submitted for virological diagnosis by polymerase chain reaction (PCR) and/or virus isolation. Thirty seven outbreaks/cases of vesicular disease were notified in cattle of 25 counties; in 33 cases the animals presented lesions clinically compatible with poxviruses. The etiology of 25 out of 33 outbreaks/cases was confirmed as poxviruses by PCR and/or viral isolation: 13 as bovine vaccinia virus (VACV), six as pseudocowpox virus (PCPV), five as bovine papular stomatitis virus (BPSV) and one coinfection (VACV and an Orf virus-like parapoxvirus). The laboratory confirmed that cases occurred mainly in dairy cattle (19/25) and during the dry season (22/25). In adult cattle, gross changes were observed mainly in the teats and udder and included vesicles, ulcers, crusts, papules and scars and varied of type, severity and affected region, depending on the poxvirus species. In calves, the main lesions were ulcers in the mouth and muzzle. Zoonotic lesions compatible with poxvirus infections were observed for all diagnosed poxviruses, affecting especially the hands of milkers and other farm workers. Our data demonstrate the sanitary and economic relevance of these diseases and the wide circulation of different poxviruses in cattle from GO.


Foi realizado um estudo retrospectivo das infecções por poxvírus diagnosticadas em bovinos do estado de Goiás (GO), entre 2010 e 2018. Todos os casos foram investigados pela Agência Goiana de Defesa Agropecuária (Agrodefesa). Foram revisados formulários técnicos e protocolos de laboratórios de diagnóstico veterinário. Na maioria dos casos, amostras de tecidos orais ou cutâneos e/ou swabs foram encaminhadas para diagnóstico virológico. Foram notificados 37 surtos/casos de doença vesicular em bovinos em 25 municípios; em 33 casos os animais apresentavam lesões clinicamente compatíveis com poxvírus. A etiologia de 25 de 33 surtos/casos foi confirmada como poxvírus por PCR e/ou isolamento viral: 13 como vírus vaccínia (VACV), seis como vírus pseudocowpox (PCPV), cinco como vírus da estomatite papular bovina (BPSV) e um caso de coinfecção (VACV e um parapoxvírus semelhante ao Orf vírus). Os casos confirmados laboratorialmente ocorreram principalmente em bovinos leiteiros (19/25) e durante a estação seca (22/25). Em bovinos adultos, alterações macroscópicas foram observadas principalmente nas tetas e úbere e incluíram vesículas, úlceras, crostas, pápulas e cicatrizes e variaram quanto ao tipo, gravidade e região afetada, dependendo da espécie do poxvírus. Em bezerros, as principais lesões foram úlceras na boca e focinho. Lesões zoonóticas compatíveis com infecção por poxvírus foram observadas em todas as poxviroses diagnosticadas, afetando principalmente as mãos dos ordenhadores e outros trabalhadores rurais. Nossos dados demonstram a relevância sanitária e econômica dessas doenças e a ampla circulação de diferentes poxvírus em bovinos de GO.


Assuntos
Humanos , Animais , Bovinos , Infecções por Poxviridae/diagnóstico , Infecções por Poxviridae/epidemiologia , Infecções por Poxviridae/patologia , Parapoxvirus/isolamento & purificação , Vaccinia virus/isolamento & purificação , Vírus da Pseudovaríola das Vacas/isolamento & purificação , Coinfecção/veterinária , Zoonoses Virais
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