qPCR assay for the detection of pseudocowpox virus.

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.

Identification of Clade E Avipoxvirus in Brazil.

Avian poxvirus (APV) is an enveloped double-stranded DNA virus that affects many domestic and wild birds worldwide. APVs are classified into three clades (A to C), represented by fowlpox (FP) virus (clade A), canarypox virus (clade B), and psittacinepox virus (clade C), although two additional clades (D and E) have been proposed. In this study, a tumorlike skin lesion found in a domestic fowl was submitted for molecular diagnosis of Avipoxvirus by PCR and sequencing. The phylogenetic analysis revealed that the amplified segment of the corelike 4b protein and polymerase genes clustered in clade E. The APVs in clade E were previously reported from outbreaks in Hungary (flock of turkeys) and in Mozambique (layer chickens), associated with a possible vaccine failure to protect against clade E viruses. To our knowledge, this report is the first identification of clade E in this country, providing new information about host range and genetic diversity of APVs in Brazil, and may represent a potential risk of FP disease outbreaks in commercial poultry.

Reporte de caso- Identificación del Avipoxvirus clado E en Brasil. El poxvirus aviar (APV) es un virus de ADN bicatenario envuelto que afecta a muchas aves domésticas y silvestres en todo el mundo. Los poxvirus aviares se clasifican en tres clados (A, B y C), representados por el virus de la viruela aviar (FP) (clado A), el virus de la viruela del canario (clado B) y el virus de la viruela de los psitácidos (clado C), aunque dos clados adicionales (D y E) han sido propuestos. En este estudio, una lesión cutánea similar a un tumor encontrada en una gallina doméstica fue sometida a diagnóstico molecular de Avipoxvirus por PCR y secuenciación. El análisis filogenético reveló que el segmento amplificado de los genes de la proteína del centro 4b y de la polimerasa se agruparon en el clado E. Los poxvirus aviares en el clado E se reportaron previamente de brotes en Hungría (parvada de pavos) y en Mozambique (gallinas de postura), asociados con una posible falla de la vacuna para proteger contra los virus del clado E. De acuerdo con el conocimiento de los autores, este informe es la primera identificación del clado E en este país, brindando nueva información sobre el rango de hospedadores y la diversidad genética de poxvirus aviares en Brasil, y puede representar un riesgo potencial de brotes de viruela aviar en aves comerciales.

Late development of pustular, erosive lesions in the muzzle of calves inoculated with Pseudocowpox virus.

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.

Poxviruses diagnosed in cattle from Distrito Federal, Brazil (2015-2018).

A retrospective study of officially diagnosed poxvirus infections in cattle in Distrito Federal (DF), Brazil, between 2015 and 2018 was performed. All cases were investigated by the DF Official Veterinary Service. In the most cases, samples of oral, cutaneous (teats, udder) or foot lesions were submitted to molecular diagnosis by PCR. In approximately 70% of the cases, additional samples were also submitted for histopathology. Ninety-three out of 2,467 clinically examined cattle (from 385 farms) presented suggestive and/or compatible lesions with poxviruses. Fifty-two out of these 93 cases were confirmed as poxviruses: 27 vaccinia virus (VACV), 9 pseudocowpox virus (PCPV), 8 bovine papular stomatitis virus (BPSV), 5 coinfection by PCPV and BPSV and 3 unidentified parapoxvirus. The clinical cases were observed in farms with different exploration (beef, dairy or mixed) from 9 out of 30 administrative regions of DF. Gross findings consisted of papules, vesicles, ulcers, scabs and scars and varied of type, severity and affected tissue, according to the detected virus. A single human case was observed associated with a BPSV infection. Histologically, the lesions were very similar, independently of the detected poxvirus, and included mild to moderate, superficial, multifocal inflammatory infiltrate of lymphocytes, plasma cells, macrophages and/or neutrophils, with acanthosis and parakeratotic hyperkeratosis, usually associated with serous content, cellular debris and spongiosis. In the ulcerated lesions, there were focally extensive areas of necrosis with severe infiltrate of neutrophils in the adjacent connective tissue. Few to moderate amount of 4- to 8-µm eosinophilic inclusion bodies were observed in the cytoplasm of keratinocytes in 6 cases (2 of VACV, 2 of PCPV and 2 of PCPV/BPSV coinfection). Data of the current study demonstrate the wide circulation of different poxviruses in cattle from DF.

Here, There, and Everywhere: The Wide Host Range and Geographic Distribution of Zoonotic Orthopoxviruses.

The global emergence of zoonotic viruses, including poxviruses, poses one of the greatest threats to human and animal health. Forty years after the eradication of smallpox, emerging zoonotic orthopoxviruses, such as monkeypox, cowpox, and vaccinia viruses continue to infect humans as well as wild and domestic animals. Currently, the geographical distribution of poxviruses in a broad range of hosts worldwide raises concerns regarding the possibility of outbreaks or viral dissemination to new geographical regions. Here, we review the global host ranges and current epidemiological understanding of zoonotic orthopoxviruses while focusing on orthopoxviruses with epidemic potential, including monkeypox, cowpox, and vaccinia viruses.

NOVEL POXVIRAL INFECTION IN THREE FINCH SPECIES ILLEGALLY IMPORTED INTO TRINIDAD, WEST INDIES, WITH IMPLICATIONS FOR NATIVE BIRDS.

Oryzoborus angolensis (Lesser Seed-Finch), Oryzoborus crassirostris (Large-billed Seed-Finch), and Sporophila intermedia (Grey Seedeater) are finch species native to the Caribbean island of Trinidad. These species are locally trapped and kept for their song, but with declining native populations, enthusiasts have turned to illegally importing birds from the South American mainland. The smuggling of wild birds from South America poses significant disease risks to the native bird species of Trinidad. Herein we describe the first case of poxviral infection in these illegally imported birds in Trinidad and partial genome sequence of the causative agent. Phylogenetic analysis of the 4b core protein sequence indicated that the avian poxvirus identified was most closely related to a 2012 avian pox sequence from Brazil, with 96.2% and 98.1% identity at the nucleotide and amino acid level.

Putative parapoxvirus-associated foot disease in the endangered huemul deer (Hippocamelus bisulcus) in Bernardo O'Higgins National Park, Chile.

The huemul (Hippocamelus bisulcus) is an endangered cervid endemic to southern Argentina and Chile. Here we report foot lesions in 24 huemul from Bernardo O'Higgins National Park, Chile, between 2005 and 2010. Affected deer displayed variably severe clinical signs, including lameness and soft tissue swelling of the limbs proximal to the hoof or in the interdigital space, ulceration of the swollen tissues, and some developed severe proliferative tissue changes that caused various types of abnormal wear, entrapment, and/or displacement of the hooves and/or dewclaws. Animals showed signs of intense pain and reduced mobility followed by loss of body condition and recumbency, which often preceded death. The disease affected both genders and all age categories. Morbidity and mortality reached 80% and 40%, respectively. Diagnostics were restricted to a limited number of cases from which samples were available. Histology revealed severe papillomatous epidermal hyperplasia and superficial dermatitis. Electron microscopy identified viral particles consistent with viruses in the Chordopoxvirinae subfamily. The presence of parapoxvirus DNA was confirmed by a pan-poxvirus PCR assay, showing high identity (98%) with bovine papular stomatitis virus and pseudocowpoxvirus. This is the first report of foot disease in huemul deer in Chile, putatively attributed to poxvirus. Given the high morbidity and mortality observed, this virus might pose a considerable conservation threat to huemul deer in Chilean Patagonia. Moreover, this report highlights a need for improved monitoring of huemul populations and synergistic, rapid response efforts to adequately address disease events that threaten the species.

PREVALENCE OF ANTIBODIES TO ORTHOPOXVIRUS IN WILD CARNIVORES OF NORTHWESTERN CHIHUAHUA, MEXICO.

The distribution of orthopoxviruses (OPXVs) across the North American continent is suggested to be widespread in a wide range of mammalian hosts on the basis of serosurveillance studies. To address the question of whether carnivores in northwestern Mexico are exposed to naturally circulating OPXVs, wild carnivores were collected by live trapping within four different habitat types during fall of 2013 and spring of 2014 within the Janos Biosphere Reserve in northwestern Chihuahua, Mexico. A total of 51 blood samples was collected for testing. Anti-OPXV immunoglobulin G enzymelinked immunosorbent assay, western blot, and rapid fluorescent focus inhibition test (RFFIT) assays were conducted. About 47% (24/51) of the carnivores tested were seropositive for anti-OPXV binding antibodies and had presence of immunodominant bands indicative of OPXV infection. All samples tested were negative for rabies virus neutralizing antibodies by RFFIT, suggesting that the OPXV antibodies were due to circulating OPXV, and not from exposure to oral rabies vaccine (vacciniavectored rabies glycoprotein vaccine) bait distributed along the US-Mexico border. Our results indicated that there may be one or more endemic OPXV circulating within six species of carnivores in northwestern Mexico.

Bayesian Phylogenetic Analysis of Avipoxviruses from North American Wild Birds Demonstrates New Insights into Host Specificity and Interspecies Transmission.

Avian pox is commonly diagnosed in a variety of North American wild and domestic birds, yet little is known about the evolutionary relationships among the causative poxviruses. This study aimed to determine the phylogenetic relationships among isolates identified in different avian host species to better characterize the host range of specific viral strains and compare the genetic variability within and between viral clades. Skin lesions grossly and microscopically consistent with poxvirus infection from 82 birds collected in Canada, the United States, and the U.S. Virgin Islands were included in this study. A total of 12 avian species were represented; the most common species sampled were wild turkeys (Meleagris gallopavo), mourning doves (Zenaida macroura), and American crows (Corvus brachyrhynchos). Poxvirus samples from these birds were genotyped using PCR that targeted the 4b core protein gene followed by amplicon sequencing. Bayesian phylogenetic analyses of these viruses, in conjunction with publicly available sequences, representing avipoxvirus strains from six continents revealed statistically significant monophyletic clades based on genetic distances of sequences within and between observed clades. Genetic variation within the fowlpox clade was low compared to the canarypox clade. Host and geographic origins of viral isolates revealed overall clustering of viral strains within avian species, with a few exceptions. No genetic differences were observed between viruses from Canada and the United States within individual species. These results are novel in their characterization and comparison of the phylogenetic relationships of poxvirus isolates in wild bird species from North America. Further, we provide new data on the level of host specificity and specific strains circulating in North America.

El análisis filogenético bayesiano de los avipoxvirus de las aves silvestres de América del Norte demuestra nuevos conocimientos sobre la especificidad del huésped y la transmisión interespecífica. La viruela aviar se diagnostica comúnmente en una variedad de aves silvestres y domésticas de América del Norte, pero se sabe poco sobre las relaciones evolutivas entre los poxvirus. Este estudio tuvo como objetivo determinar las relaciones filogenéticas entre aislamientos identificados en diferentes especies de hospedadores aviares para caracterizar mejor el rango de hospedadores de cepas virales específicas y comparar la variabilidad genética dentro y entre los clados virales. Se incluyeron en este estudio lesiones cutáneas que eran consistentes macro y microscópicamente con la infección por poxvirus de 82 aves recolectadas en Canadá, Estados Unidos y las Islas Vírgenes de los Estados Unidos. Un total de 12 especies de aves fueron representadas; las especies más comunes en la muestra fueron los pavos silvestres (Meleagris gallopavo), huilota común (Zenaida macroura) y cuervos americanos (Corvus brachyrhynchos). Las muestras de poxvirus de estas aves fueron genotipadas mediante PCR que se enfocó en el gene de la proteína central 4b seguido de secuenciación de amplicón. Los análisis filogenéticos bayesianos de estos virus, junto con las secuencias disponibles públicamente, que representan cepas de avipoxvirus de seis continentes revelaron clados monofiléticos estadísticamente significativos basados en distancias genéticas de las secuencias dentro y entre los clados observados. La variación genética dentro del clado de la viruela del pollo fue baja en comparación con el clado de virus de canario. El huésped y los orígenes geográficos de los aislamientos virales revelaron un agrupamiento general de cepas virales dentro de las especies aviares, con algunas excepciones. No se observaron diferencias genéticas entre los virus de Canadá y los Estados Unidos dentro de las especies individuales. Estos resultados son novedosos en la caracterización y comparación de las relaciones filogenéticas de los aislados de poxvirus en especies de aves silvestres de América del Norte. Además, se proporcionan nuevos datos sobre el nivel de especificidad del huésped y las cepas específicas que circulan en América del Norte. Key words: Bayesian analysis, mourning dove, phylogenetic, poxvirus, sequencing, wild turkey, 4b gene.

Outbreak of Systemic Avian Pox in Canaries (Serinus canaria domestica) Associated with the B1 Subgroup of Avian Pox Viruses.

This study reports an outbreak of avian pox in a quarantine of canaries imported from Europe, with a mortality of 30% and clinical signs of dyspnea and blepharoconjunctivitis. During necropsy, beak cyanosis, serous blepharitis, caseous sinusitis, oropharyngitis, tracheitis, pulmonary edema, pneumonia, fibrinous airsacculitis, and splenomegaly were observed. Microscopically, edema, epithelial hyperplasia, hydropic degeneration, and vacuolated eosinophilic intracytoplasmic inclusion bodies were found; similar lesions were observed in the thymus, spleen, and other organs. The virus was isolated in chicken embryos, and it was identified and characterized using a sequence of 913 nucleotides of the DNA polymerase gene. Pathologic characteristics and molecular biology indicate the systemic presence of avian pox associated with an avipoxvirus of the B1 subgroup. Additionally, other lesions associated with Aspergillus sp., Macrorhabdus ornithogaster, and Isospora sp. were found, which could contribute to the high mortality. Canarypox virus should be considered a differential diagnosis in cases of dyspnea and high mortality in canary flocks.

Reporte de caso- Brote de viruela aviar sistémica en canarios (Serinus canaria domestica) asociado con el virus de la viruela aviar subgrupo B1. En este estudio se reporta un brote de viruela aviar en unos canarios importados de Europa que estaban bajo cuarentena, con una mortalidad del 30% y signos clínicos de disnea y blefaroconjuntivitis. Durante la necropsia, se observó cianosis de pico, blefaritis serosa, sinusitis caseosa, orofaringitis, traqueítis, edema pulmonar, neumonía, fibrinosa respiratoria y esplenomegalia. Microscópicamente, se encontró edema, hiperplasia epitelial, degeneración hidrópica y cuerpos de inclusión intracitoplásmicos eosinófilos vacuolados; Se observaron lesiones similares en el timo, el bazo y otros órganos. El virus se aisló en embriones de pollo, se identificó y caracterizó analizando una secuencia de 913 nucleótidos del gene de la polimerasa de ADN. Las características patológicas y la biología molecular indican la presentación sistémica de la viruela aviar asociada a un avipoxvirus del subgrupo B1. Además, se encontraron otras lesiones asociadas con Aspergillus sp., Macrorhabdus ornithogaster y Isopora sp., lo que pudo contribuir a la alta mortalidad. El virus de la viruela del canario debe considerarse un diagnóstico diferencial en casos de disnea y alta mortalidad en parvadas de canarios.

Spread of poxviruses in livestock in Brazil associated with cases of double and triple infection.

The objective of this work is to describe the distribution of outbreaks of vaccinia virus (VACV), pseudocowpox virus (PCPV), and bovine papular stomatitis virus (BSPV) in Brazil. The Official Laboratory of the Brazilian Ministry of Agriculture received 89 samples from different locations in Brazil in 2015 and 2016 for diagnosis of vesicular and exanthematous disease. Poxvirus coinfections occurred in 11 out of 33 outbreaks, including the first reported triple infection by BPSV, PCPV, and VACV. This occurrence may be associated with the circulation of these viruses in Brazilian cattle.

Detection and Molecular Characterization of Zoonotic Poxviruses Circulating in the Amazon Region of Colombia, 2014.

During 2014, cutaneous lesions were reported in dairy cattle and farmworkers in the Amazon Region of western Colombia. Samples from 6 patients were analyzed by serologic and PCR testing, and results demonstrated the presence of vaccinia virus and pseudocowpox virus. These findings highlight the need for increased poxvirus surveillance in Colombia.

Detection of pseudocowpox virus in water buffalo (Bubalus bubalis) with vesicular disease in the state of São Paulo, Brazil, in 2016.

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.

A multiplex PCR for viruses associated with exanthematic and vesicular disease in cattle.

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.

Avian Pox in Native Captive Psittacines, Brazil, 2015.

To investigate an outbreak of avian pox in psittacines in a conservation facility, we examined 94 birds of 10 psittacine species, including sick and healthy birds. We found psittacine pox virus in 23 of 27 sick birds and 4 of 67 healthy birds. Further characterization is needed for these isolates.

Occurrence of Pseudocowpox virus associated to Bovine viral diarrhea virus-1, Brazilian Amazon.

In 2011, an outbreak of severe vesicular disease occurred in the state of Pará, Amazon region. Besides proliferative or verrucous lesions, cattle showed atypical clinical signs such as diarrhea and leading to death. The animals were submitted to clinical, pathological and molecular diagnosis, and laboratory tests have confirmed the presence of Pseudocowpox virus (PCPV), a Parapoxvirus genus member, and have also found Bovine viral diarrhea virus-1 (BVDV-1), probably causing persistent infection. The results of molecular diagnostics, followed by sequencing data demonstrated the circulation of both viruses (PCPV and BVDV-1) in an area previously affected by another poxvirus, as Vaccinia virus.The cocirculation between PCPV and BVDV-1 indicates a major concern for animal health because the clinical presentation can be a severe disease. This is the first detection of PCPV in the Brazilian Amazon.

Seroprevalence of Orthopoxvirus in rural Brazil: insights into anti-OPV immunity status and its implications for emergent zoonotic OPV.

BACKGROUND: Bovine vaccinia (BV) is a zoonosis caused by Vaccinia virus, a virus from Orthopoxvirus genus (OPV) that affects mainly cattle herds and humans in rural areas in Brazil. Because most studies have focused on outbreaks situations, data on BV epidemiology is limited. A cross sectional study in Brazilian rural areas during 2012-2013 was conducted to determine the neutralizing antibodies seroprevalence and risk factors for BV. METHODS: A structured questionnaire was applied to elicit demographics data and farming practices considered risk factors for BV exposure. Neutralizing anti-OPV antibodies were investigated using plaque reduction neutralization test. The neutralizing antibodies prevalence rates were calculated and the risk factor analysis was performed using multivariate logistic regression. RESULTS: Two hundred and forty participants were enrolled in this study with a prevalence of neutralizing antibodies of 30.8 % (95 % confidence interval [CI], 25.3-36.9). In multivariate analysis, age > 35 years (Odds Ratio [OR] = 18.2; CI 95 % = 7.7 - 43.2) and previous outbreak in property (OR = 3.9; C I95 % = 1.2 - 12.6) were independently associated with anti-OPV neutralizing antibodies. CONCLUSIONS: In this study, anti-OPV protective immunity (neutralizing antibody titers) was assessed in an endemic BV Brazilian rural area. Our findings indicate that epidemiological surveillance is required and should be applied by public health authorities to create interventions and/or prevention strategies to avoid viral spread causing future outbreaks among individuals who are under risk of infection.

The detection of Vaccinia virus confirms the high circulation of Orthopoxvirus in buffaloes living in geographical isolation, Marajó Island, Brazilian Amazon.

In Brazil, serologic evidence of Orthopoxvirus (OPV) circulation showed positivity around 20% in cattle, humans, monkeys and rodents. Although OPV seropositivity has been described in buffalo herds in southeastern Brazil, no Vaccinia virus (VACV) (member of genus OPV) outbreaks in buffalo herds have been described in this country. This study aimed to investigate the detection of anti-OPV antibodies and to study the OPV genome in Brazilian buffalo herds. Our results demonstrated a high OPV seropositivity in buffalo herds on Marajó Island and molecular data confirmed the circulation of VACV. The geographical isolation conditionmight be a sine qua non condition to explain our results.

Detection of multiple viral infections in cattle and buffalo with suspected vesicular disease in Brazil.

Vesicular diseases are of high importance for livestock, primarily because of foot-and-mouth disease (FMD), which is a high-morbidity disease that generates direct losses caused by low milk production, weight loss, and indirect losses because of the need for sanitary barriers. Other vesicular diseases are also of importance for livestock because of direct impacts or because their clinical signs may be confused with those of FMD. We report herein the detection of multiple infections in cattle with suspected vesicular disease in the Brazilian states of Amazonas (AM), Mato Grosso (MT), and Roraima. Thirty-seven epithelial samples from cattle and 1 sample from a buffalo were sent to the laboratory for testing for FMDV and similar disease agents. All samples from MT were positive for parapoxvirus (Pseudocowpox virus and Bovine papular stomatitis virus). In addition, 3 samples were positive for Bluetongue virus, and 5 samples were positive for Bovine herpesvirus 1 Among these samples, 1 was positive for all of these 3 agents. Only 2 samples from AM were negative for parapoxvirus. The molecular tests conducted in this study detected multiple infections, with a high prevalence of parapoxvirus.