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1.
Braz. j. vet. pathol ; 16(1): 64-70, mar. 2023. ilus, graf
Article in English | VETINDEX | ID: biblio-1425388

ABSTRACT

A postmortem study was performed on two lovebirds (Agapornis fischeri and Agapornis personatus) that had scabs in the periocular region and on the eyelid, as well as serous blepharitis. Microscopically, the eyelids showed ulcers, necrosis and serocellular crusts, severe hyperplasia of keratinocytes with eosinophilic intracytoplasmic inclusion bodies (Bollinger's bodies), bacterial colonies of gram-positive coccoid morphology and PAS-positive septate and 45° branching hyphae. The microbiological study identified the colonies as Staphylococcus spp. and Aspergillus fumigatus, respectively. Using molecular techniques, avian pox clade C was identified on the eyelid. This is the first report in Mexico of a case of avian pox in parrots associated with clade C Avipoxvirus.(AU)


Subject(s)
Animals , Poxviridae Infections/diagnosis , Agapornis/anatomy & histology , Agapornis/classification , Aspergillus fumigatus , Staphylococcus , Avipoxvirus/pathogenicity , Mexico
2.
Braz J Microbiol ; 53(2): 1077-1080, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35220553

ABSTRACT

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.


Subject(s)
Cattle Diseases , Poxviridae Infections , Animals , Cattle , Cattle Diseases/diagnosis , Pathology, Molecular , Poxviridae Infections/diagnosis , Poxviridae Infections/veterinary , Pseudocowpox Virus/genetics , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Sensitivity and Specificity
3.
Pesqui. vet. bras ; Pesqui. vet. bras;42: e07014, 2022. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1360625

ABSTRACT

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)


Subject(s)
Humans , Animals , Cattle , Vaccinia virus/isolation & purification , Parapoxvirus/isolation & purification , Pseudocowpox Virus/isolation & purification , Poxviridae Infections/diagnosis , Poxviridae Infections/pathology , Poxviridae Infections/epidemiology , Coinfection/veterinary , Viral Zoonoses
4.
Pesqui. vet. bras ; Pesqui. vet. bras;42: e07014, 2022. tab, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1487701

ABSTRACT

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.


Subject(s)
Humans , Animals , Cattle , Poxviridae Infections/diagnosis , Poxviridae Infections/epidemiology , Poxviridae Infections/pathology , Parapoxvirus/isolation & purification , Vaccinia virus/isolation & purification , Pseudocowpox Virus/isolation & purification , Coinfection/veterinary , Viral Zoonoses
5.
Avian Dis ; 64(2): 223-227, 2020 06.
Article in English | MEDLINE | ID: mdl-32550624

ABSTRACT

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.


Subject(s)
Avipoxvirus/isolation & purification , Chickens , Poultry Diseases/diagnosis , Poxviridae Infections/veterinary , Animals , Brazil , Poultry Diseases/virology , Poxviridae Infections/diagnosis , Poxviridae Infections/virology
6.
Arch Virol ; 164(12): 3059-3063, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31549301

ABSTRACT

Swine are the only known hosts of swinepox virus (SWPV), the sole member of the genus Suipoxvirus, family Poxviridae. Rapid diagnosis is recommended for appropriate interventions because of the high morbidity associated with this virus. This study describes a real-time quantitative PCR (qPCR) assay for rapid detection and quantification of SWPV. The detection limit, repeatability, reproducibility, and specificity of this assay were determined. The efficiency was 96%, and the R2 value was 0.996. The detection limit was 1 fg or 10-0.5 TCID50/50 µL. Tests showed that the greatest source of error in the SWPV qPCR assay was variation between analysts rather than different qPCR kits or equipment. All nucleic acids from other viruses or samples collected from swine were negative in the specificity test. qPCR for SWPV is a new method with tested variables that allows main sources of error in laboratory diagnosis and viral quantification to be identified.


Subject(s)
Poxviridae Infections/diagnosis , Suipoxvirus/genetics , Swine Diseases/virology , Animals , DNA, Viral/genetics , Limit of Detection , Poxviridae Infections/veterinary , Real-Time Polymerase Chain Reaction , Suipoxvirus/classification , Suipoxvirus/isolation & purification , Swine
7.
J Virol Methods ; 239: 38-41, 2017 01.
Article in English | MEDLINE | ID: mdl-27793645

ABSTRACT

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.


Subject(s)
Cattle Diseases/diagnosis , Cattle Diseases/virology , Multiplex Polymerase Chain Reaction/methods , Poxviridae Infections/veterinary , Animals , Brazil , Cattle , DNA Primers , DNA, Viral/genetics , Genes, Viral , Genome, Viral , Orthopoxvirus/genetics , Orthopoxvirus/isolation & purification , Parapoxvirus/genetics , Parapoxvirus/isolation & purification , Poxviridae Infections/diagnosis , Poxviridae Infections/virology , Pseudocowpox Virus/genetics , Pseudocowpox Virus/isolation & purification , Vaccinia virus/genetics , Vaccinia virus/isolation & purification
8.
Article in English | MEDLINE | ID: mdl-27865267

ABSTRACT

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.


Subject(s)
Bovine Virus Diarrhea-Mucosal Disease/virology , Cattle Diseases/epidemiology , Cattle Diseases/virology , Coinfection/veterinary , Disease Outbreaks/veterinary , Poxviridae Infections/veterinary , Pseudocowpox Virus/isolation & purification , Animals , Antigens, Viral/blood , Antigens, Viral/genetics , Bovine Virus Diarrhea-Mucosal Disease/diagnosis , Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Bovine Virus Diarrhea-Mucosal Disease/physiopathology , Brazil/epidemiology , Cattle , Cattle Diseases/diagnosis , Coinfection/diagnosis , Coinfection/epidemiology , Coinfection/virology , Diarrhea , Diarrhea Virus 1, Bovine Viral/genetics , Phylogeny , Poxviridae Infections/diagnosis , Poxviridae Infections/epidemiology , Poxviridae Infections/virology , Pseudocowpox Virus/genetics , Real-Time Polymerase Chain Reaction , Sequence Analysis, DNA
9.
J Vet Diagn Invest ; 28(4): 377-81, 2016 Jul.
Article in English | MEDLINE | ID: mdl-27154321

ABSTRACT

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.


Subject(s)
Bluetongue/diagnosis , Buffaloes , Cattle Diseases/diagnosis , Herpesviridae Infections/veterinary , Poxviridae Infections/veterinary , Animals , Bluetongue/virology , Bluetongue virus/isolation & purification , Brazil/epidemiology , Cattle , Cattle Diseases/virology , Herpesviridae Infections/diagnosis , Herpesviridae Infections/virology , Herpesvirus 1, Bovine/isolation & purification , Parapoxvirus/isolation & purification , Poxviridae Infections/diagnosis , Poxviridae Infections/virology
10.
PLoS One ; 6(1): e15989, 2011 Jan 13.
Article in English | MEDLINE | ID: mdl-21249151

ABSTRACT

The role of disease in regulating populations is controversial, partly owing to the absence of good disease records in historic wildlife populations. We examined birds collected in the Galapagos Islands between 1891 and 1906 that are currently held at the California Academy of Sciences and the Zoologisches Staatssammlung Muenchen, including 3973 specimens representing species from two well-studied families of endemic passerine birds: finches and mockingbirds. Beginning with samples collected in 1899, we observed cutaneous lesions consistent with Avipoxvirus on 226 (6.3%) specimens. Histopathology and viral genotyping of 59 candidate tissue samples from six islands showed that 21 (35.6%) were positive for Avipoxvirus, while alternative diagnoses for some of those testing negative by both methods were feather follicle cysts, non-specific dermatitis, or post mortem fungal colonization. Positive specimens were significantly nonrandomly distributed among islands both for mockingbirds (San Cristobal vs. Espanola, Santa Fe and Santa Cruz) and for finches (San Cristobal and Isabela vs. Santa Cruz and Floreana), and overall highly significantly distributed toward islands that were inhabited by humans (San Cristobal, Isabela, Floreana) vs. uninhabited at the time of collection (Santa Cruz, Santa Fe, Espanola), with only one positive individual on an uninhabited island. Eleven of the positive specimens sequenced successfully were identical at four diagnostic sites to the two canarypox variants previously described in contemporary Galapagos passerines. We conclude that this virus was introduced late in 1890's and was dispersed among islands by a variety of mechanisms, including regular human movements among colonized islands. At present, this disease represents an ongoing threat to the birds on the Galapagos Islands.


Subject(s)
Avipoxvirus , Bird Diseases/transmission , Passeriformes/virology , Poxviridae Infections/veterinary , Animals , Bird Diseases/diagnosis , Ecuador , Humans , Poxviridae Infections/diagnosis , Time Factors
11.
J Clin Virol ; 48(1): 69-72, 2010 May.
Article in English | MEDLINE | ID: mdl-20207192

ABSTRACT

BACKGROUND: Occupational exanthematic diseases represent an important cause of public health impact and economical losses. Among the viral exanthematic diseases, two caused by poxviruses are noteworthy: the bovine vaccinia (BV), caused by the Vaccinia virus (VACV); and the milker's nodule, in which the agent is the Pseudocowpox virus (PCPV). Both agents are zoonotic and have been associated with several cases of bovine infection. In Brazilian rural areas BV has been highly prevalent, particularly in milk herds. Farmers, milkers and their close contacts developed lesions on the hands, forearms, legs and face accompanied by several systemic symptoms. Although VACV and PCPV present with similar epidemiological and transmission patterns, no VACV and PCPV co-infection cases have to date been described. OBJECTIVES: To describe the first case of zoonotic VACV and PCVP co-infection, based on serological and molecular methods. STUDY DESIGN AND RESULTS: In this work we report a case of a Brazilian rural worker who presented with a large severely ulcerated-pustule skin lesion, associated with fever, headache, malaise, myalgia and axillary, inguinal and cervical limphadenopathy. The worker declared occupational contact with cattle that had notable injuries on their teats. Human and bovine clinical samples were collected and submitted to serological and molecular tests. PCR and phylogenetic analysis revealed the presence of VACV DNA and PCPV DNA in the patient's lesion. Serological tests indicated anti-VACV neutralizing antibodies and molecular assays showed the presence of VACV and PCPV DNA in the patient sera. VACV and PCPV also were detected in dairy cattle. CONCLUSION: Together, these results indicate a case of zoonotic VACV/PCPV co-infection. Epidemiological surveillance and appropriate medical treatment are essential for the control of both diseases, especially in the most severe cases, as described in the present study.


Subject(s)
Poxviridae Infections/virology , Pseudocowpox Virus/genetics , Vaccinia virus/genetics , Vaccinia/virology , Zoonoses/virology , Animals , Brazil , Cattle , Fingers/pathology , Fingers/virology , Humans , Male , Phylogeny , Poxviridae Infections/diagnosis , Skin/pathology , Skin/virology , Young Adult
12.
J Med Virol ; 82(4): 692-9, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20166167

ABSTRACT

Orthopoxvirus (OPV) has been associated with worldwide exanthematic outbreaks, which have resulted in serious economic losses as well as impact on public health. Although the current classical and molecular methods are useful for the diagnosis of OPV, they are largely inaccessible to unsophisticated clinical laboratories. The major reason for the inaccessibility is that they require both virus isolation and DNA manipulation. In this report, a rapid, sensitive and low-cost semi-nested PCR method is described for the detection of OPV DNA directly from clinical specimens. A set of primers was designed to amplify the conserved OPV vgf gene. The most useful thermal and chemical conditions were selected and minimum non-inhibitory dilutions were determined. More than 100 Brazilian Vaccinia virus (VACV) field clinical specimens were tested using this semi-nested PCR in order to confirm its applicability. Cowpox virus was also detected by PCR from the ear scabs of scarified Balb/c mice. In addition, the method was highly sensitive for the detection of VACV DNA in murine blood and excreta, which are among the suggested reservoirs of OPV. Together, these data suggest that semi-nested PCR can be used for initial screening for OPV and as a routine diagnostic laboratory method.


Subject(s)
Clinical Laboratory Techniques/methods , Orthopoxvirus/isolation & purification , Polymerase Chain Reaction/methods , Poxviridae Infections/diagnosis , Poxviridae Infections/veterinary , Animals , Brazil , Cattle , Cattle Diseases/diagnosis , Cattle Diseases/virology , DNA Primers/genetics , DNA, Viral/genetics , Disease Models, Animal , Genes, Viral , Humans , Male , Mice , Mice, Inbred BALB C , Orthopoxvirus/genetics , Poxviridae Infections/virology , Sensitivity and Specificity
13.
Pesqui. vet. bras ; Pesqui. vet. bras;29(7): 509-514, July 2009. ilus
Article in English | LILACS | ID: lil-526791

ABSTRACT

In the present study we investigated the presence of infections by vaccinia-like viruses in dairy cattle from 12 counties in the state of Rio de Janeiro in the last 9 years. Clinical specimens were collected from adult animals with vesicular/pustular lesions mainly in the udder and teats, and from calves with lesions around the nose and mouth. A plaque reduction neutralization test (PRNT) was applied to search for antibodies to Orthopoxvirus; the vesicular/pustular fluids and scabs were examined by PCR, electron microscopy (EM) and by inoculation in VERO cells for virus isolation. Antibodies to Orthopoxvirus were detected in most cases. The PCR test indicated a high nucleotide homology among the isolates and the vaccinia viruses (VACV) used as controls. By EM, typical orthopoxvirus particles were observed in some specimens. The agents isolated in tissue culture were confirmed as vaccinia-like viruses by EM and PCR. The HA gene of the vaccinia-like Cantagalo/IOC virus isolated in our laboratory was sequenced and compared with other vaccinia-like isolates, showing high homology with the original Cantagalo strain, both strains isolated in 1999 from dairy cattle. Antibodies to Orthopoxvirus were detected in one wild rodent (genus Akodon sp.) collected in the northwestern region of the state, indicating the circulation of poxvirus in this area. Nonetheless, PCR applied to tissue samples collected from the wild rodents were negative. Vesicular/pustular lesions in people in close contact with animals have been also recorded. Thus, the vaccinia-like virus infections in cattle and humans in the state seem to be an expanding condition, resulting in economic losses to dairy herds and leading to transient incapacitating human disease. Therefore, a possible immunization of the dairy cattle in the state should be carefully evaluated.


Neste estudo avaliou-se a presença de infecções por vírus semelhantes ao vírus vaccínia (VACV) em gado leiteiro em 12 municípios no estado do Rio de Janeiro, ao longo dos últimos nove anos. Amostras clínicas foram coletadas de animais com vesículas, pústulas e crostas no úbere e tetas, e da região do nariz e da cavidade oral de bezerros. Um teste de neutralização viral por redução de placas foi desenvolvido para investigar a presença de anticorpos contra Orthopoxvirus. Os fluidos de vesículas / pústulas e as crostas foram testadas por PCR, microscopia eletrônica (ME) e por inoculação em células VERO para isolamento viral. Anticorpos contra Orthopoxvirus foram detectados na grande maioria dos animais. O teste de PCR demonstrou homologia entre os vírus isolados e amostras de vírus vaccínia usados como controles. Na ME, partículas típicas de Orthopoxvirus foram observadas em vários espécimes analisados. Os vírus isolados em cultivo celular foram confirmados como Orthopoxvirus por PCR e ME. O gene HA da amostra Cantagalo/IOC isolada em nosso laboratório foi seqüenciado e comparado com outras amostras semelhantes ao vaccínia, mostrando uma alta homologia com a amostra original Cantagalo, tendo sido as duas amostras isoladas em 1999 de gado leiteiro. Anticorpos para Orthopoxvirus foram detectados em um roedor silvestre do gênero Akodon sp. coletado na região noroeste do estado, sugerindo uma circulação de poxvírus na natureza. No entanto, os testes de PCR aplicados a tecidos de roedores silvestres foram negativos. Infecções vesiculares / pustulares em humanos que mantinham contato com os animais afetados também foram relatadas. Assim, infecções por amostras semelhantes ao vírus VACV em bovinos e em humanos parecem em expansão no estado, gerando perdas econômicas em animais e quadros de doença incapacitante temporária em pacientes humanos. Dessa forma, a possibilidade da imunização do gado leiteiro no estado deve ser devidamente avaliada.


Subject(s)
Animals , Poxviridae Infections/complications , Poxviridae Infections/diagnosis , Poxviridae Infections/epidemiology , Poxviridae Infections/veterinary , Microscopy, Electron/methods , Orthopoxvirus/isolation & purification , Polymerase Chain Reaction/methods , Arvicolinae , Cattle , Brazil/epidemiology , Neutralization Tests/methods , Neutralization Tests/veterinary
14.
Pesqui. vet. bras ; 29(7): 509-514, 2009. ilus
Article in English | VETINDEX | ID: vti-567

ABSTRACT

In the present study we investigated the presence of infections by vaccinia-like viruses in dairy cattle from 12 counties in the state of Rio de Janeiro in the last 9 years. Clinical specimens were collected from adult animals with vesicular/pustular lesions mainly in the udder and teats, and from calves with lesions around the nose and mouth. A plaque reduction neutralization test (PRNT) was applied to search for antibodies to Orthopoxvirus; the vesicular/pustular fluids and scabs were examined by PCR, electron microscopy (EM) and by inoculation in VERO cells for virus isolation. Antibodies to Orthopoxvirus were detected in most cases. The PCR test indicated a high nucleotide homology among the isolates and the vaccinia viruses (VACV) used as controls. By EM, typical orthopoxvirus particles were observed in some specimens. The agents isolated in tissue culture were confirmed as vaccinia-like viruses by EM and PCR. The HA gene of the vaccinia-like Cantagalo/IOC virus isolated in our laboratory was sequenced and compared with other vaccinia-like isolates, showing high homology with the original Cantagalo strain, both strains isolated in 1999 from dairy cattle. Antibodies to Orthopoxvirus were detected in one wild rodent (genus Akodon sp.) collected in the northwestern region of the state, indicating the circulation of poxvirus in this area. Nonetheless, PCR applied to tissue samples collected from the wild rodents were negative. Vesicular/pustular lesions in people in close contact with animals have been also recorded. Thus, the vaccinia-like virus infections in cattle and humans in the state seem to be an expanding condition, resulting in economic losses to dairy herds and leading to transient incapacitating human disease. Therefore, a possible immunization of the dairy cattle in the state should be carefully evaluated.(AU)


Neste estudo avaliou-se a presença de infecções por vírus semelhantes ao vírus vaccínia (VACV) em gado leiteiro em 12 municípios no estado do Rio de Janeiro, ao longo dos últimos nove anos. Amostras clínicas foram coletadas de animais com vesículas, pústulas e crostas no úbere e tetas, e da região do nariz e da cavidade oral de bezerros. Um teste de neutralização viral por redução de placas foi desenvolvido para investigar a presença de anticorpos contra Orthopoxvirus. Os fluidos de vesículas / pústulas e as crostas foram testadas por PCR, microscopia eletrônica (ME) e por inoculação em células VERO para isolamento viral. Anticorpos contra Orthopoxvirus foram detectados na grande maioria dos animais. O teste de PCR demonstrou homologia entre os vírus isolados e amostras de vírus vaccínia usados como controles. Na ME, partículas típicas de Orthopoxvirus foram observadas em vários espécimes analisados. Os vírus isolados em cultivo celular foram confirmados como Orthopoxvirus por PCR e ME. O gene HA da amostra Cantagalo/IOC isolada em nosso laboratório foi seqüenciado e comparado com outras amostras semelhantes ao vaccínia, mostrando uma alta homologia com a amostra original Cantagalo, tendo sido as duas amostras isoladas em 1999 de gado leiteiro. Anticorpos para Orthopoxvirus foram detectados em um roedor silvestre do gênero Akodon sp. coletado na região noroeste do estado, sugerindo uma circulação de poxvírus na natureza. No entanto, os testes de PCR aplicados a tecidos de roedores silvestres foram negativos. Infecções vesiculares / pustulares em humanos que mantinham contato com os animais afetados também foram relatadas. Assim, infecções por amostras semelhantes ao vírus VACV em bovinos e em humanos parecem em expansão no estado, gerando perdas econômicas em animais e quadros de doença incapacitante temporária em pacientes humanos. Dessa forma, a possibilidade da imunização do gado leiteiro no estado deve ser devidamente avaliada.(AU)


Subject(s)
Animals , Orthopoxvirus/isolation & purification , Poxviridae Infections/complications , Poxviridae Infections/diagnosis , Poxviridae Infections/epidemiology , Poxviridae Infections/veterinary , Microscopy, Electron/methods , Polymerase Chain Reaction/methods , Neutralization Tests/methods , Neutralization Tests/veterinary , Cattle , Arvicolinae , Brazil/epidemiology
15.
Folia dermatol. peru ; 17(3): 133-136, sept.-dic. 2006. ilus
Article in Spanish | LILACS | ID: lil-475626

ABSTRACT

El nódulo de los ordeñadores es una afección viral benigna que por lo general se presenta como un nódulo en la mano de personas que manipulan ganado bovino infectado. Reportamos el caso de una mujer de 36 años con infrecuente localización facial consecutivo a cornada de vaca. Los rasgos clínicos característicos, la autorresolución del cuadro, así como los cambios citopáticos virales en la biopsia fueron significativos para establecer el diagnóstico. Es una enfermedad que podemos encontrarla en zonas periurbanas de nuestro país y factible de condicionar error diagnóstico.


Subject(s)
Humans , Female , Adult , Poxviridae Infections/diagnosis , Poxviridae Infections/therapy , Pseudocowpox Virus
16.
J. venom. anim. toxins incl. trop. dis ; J. venom. anim. toxins incl. trop. dis;11(2): 102-108, May-Aug. 2005. ilus
Article in English | LILACS | ID: lil-402359

ABSTRACT

Sheep and cattle parapoxviruses cause in human beings diseases of very similar aspect, named orf and milker's nodules, respectively. These infections are generically called farmyard pox. In the present article, we show the epidemiological, clinical, and histopathological aspects, as well as the treatment of these two viral diseases that are very similar, being differentiated only by their epidemiological aspects


Subject(s)
Humans , Animals , Male , Female , Ecthyma, Contagious/diagnosis , Ecthyma, Contagious/pathology , Poxviridae Infections/diagnosis , Poxviridae Infections/epidemiology , Poxviridae Infections/pathology , Poxviridae Infections/therapy , Cattle , Sheep
18.
An. bras. dermatol ; An. bras. dermatol;72(5): 477-80, set.-out. 1997. tab, ilus
Article in Portuguese | LILACS | ID: lil-217802

ABSTRACT

Os autores apresentam estudo de dez pacientes portadores de nódulos dos ordenhadores, com idades variando de 13 a 48 anos, sendo oito do sexo masculino e dois do feminino, todos com atividades relacionadas ao contato ou manuseio de animais ou derivados contaminados. O número de lesöes variou de um a três, todas localizadas nas mäos e nos dedos. Dois pacientes apresentaram fenômenos associados, febre em um e no outro linfangite e erupçäo micropapulosa do membro superior direito. Näo foram realizados tratamentos, excluído o sintomático, havendo regressäo espontânea das lesoes


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Animals , Cattle , Lymphangitis/diagnosis , Poxviridae Infections , Pseudocowpox Virus , Skin Diseases, Viral , Diagnosis, Differential , Granuloma, Pyogenic/diagnosis , Poxviridae Infections/complications , Poxviridae Infections/diagnosis , Poxviridae Infections/pathology , Poxviridae Infections/transmission , Sporotrichosis/diagnosis
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