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A campilobacteriose é uma doença causada por bactérias do gênero Campylobacter, responsáveis por uma das principais zoonoses de caráter global. As espécies de Campylobacter comumente isoladas de amostras fecais de cães e gatos são C. upsaliensis, C. helveticus e C. jejuni e, embora a maioria destes animais apresentem infecções subclínicas, alguns desenvolvem enterite leve a moderada. Animais jovens, mantidos sob alta densidade populacional e/ou convivendo com outros animais com doenças concomitantes são especialmente predispostos à infecção e ao desenvolvimento dos sinais clínicos. O contato com cães e gatos é um fator de risco reconhecido para a campilobacteriose humana, sendo que infecções por Campylobacter e suas complicações, como a Síndrome de Guillain-Barré, podem causar crescente morbidade em seres humanos suscetíveis. Portanto, indivíduos imunossuprimidos, que vivem ou trabalham em contato próximo com estes animais, devem ser informados dos micro-organismos zoonóticos que podem estar presentes. A compreensão da epidemiologia, patogenicidade, fatores de risco e medidas de prevenção e controle da campilobacteriose ampliou nos últimos anos, juntamente com o reconhecimento de novas espécies e diversidade genômica, atribuídos em grande parte à melhoria da tecnologia, permitindo análises comparativas destes micro-organismos. Embora tenha ocorrido progresso no entendimento do potencial patogênico de Campylobacter spp., o incremento nas pesquisas envolvendo este agente bacteriano ainda precisa ser realizado. Palavras-chave: Campylobacter. Zoonose. Diarreia. Abstract Campylobacteriosis is a disease caused by bacteria of the genus Campylobacter, representing one of the main global zoonoses. Campylobacter species commonly isolated from dogs’ and cats’ fecal samples are C. upsaliensis, C. helveticus, and C. jejuni, and although most of these animals have subclinical infections, some develop mild to moderate enteritis. Young animals kept under high population density and / or living with other animals presenting concomitant diseases are especially predisposed to infection and to the development of clinical signs. Contact with infected dogs and cats is a recognized risk factor for human campylobacteriosis, and infections by Campylobacter and its complications, such as Guillain-Barre syndrome, can cause increasing morbidity in susceptible humans. Therefore, immunosuppressed individuals living or working in close contact with these animals should be informed of the zoonotic microorganisms that may be present. The understanding of epidemiology, pathogenicity, risk factors, preventive and control measures for campylobacteriosis has expanded in recent years along with the recognition of new species and genomic diversity, largely attributed to the technology improvement, allowing comparative analyses of these microorganisms. Although progress has been made in understanding the pathogenic potential of Campylobacter spp., the increase in research involving this bacterial agent still needs to be carried out. Keywords: Campylobacter. Zoonosis. Diarrhea.

Hendra henipavírus (HeV) e Nipah henipavírus (NiV) são membros do gênero Henipavirus, pertencente à família Paramyxoviridae, sendo classificados como patógenos de nível de biossegurança 4, em função de sua alta capacidade em causar doença letal em seres humanos associada à constituição genética única, carência de terapia e profilaxia específicas. O reservatório natural destes vírus são os morcegos pertencentes ao gênero Pteropus, encontrados em regiões que se estendem do Pacífico Ocidental à Costa Leste da África. O desmatamento é um dos responsáveis pela saída dos morcegos de seus nichos ecológicos e aproximação de fazendas e vilarejos. Novos casos de infecção pelo HeV em cavalos continuam ocorrendo na Austrália, enquanto o NiV é responsável por surtos anuais em humanos, desde 2001, na Índia e Bangladesh. O NiV, em particular, possui vários recursos que destacam seu potencial como ameaça pandêmica, incluindo sua capacidade de infectar humanos diretamente, a partir de reservatórios naturais, além de uma capacidade limitada de transmissão entre seres humanos. Apesar disso, atualmente, pouco se sabe sobre os mecanismos pelos quais os morcegos abrigam vírus capazes de causar doenças tão graves em outros mamíferos terrestres. A presente revisão traz informações relevantes para o entendimento sobre a epidemiologia destas viroses, a patogenia nas espécies suscetíveis, bem como a importância destes vírus nas espécies domésticas, principalmente, nos equídeos. Palavras-chave: Hendra Vírus. Nipah Vírus. Morcegos. Equinos. Suínos. Abstract Hendra henipavirus (HeV) and Nipah henipavirus (NiV) are members of the genus Henipavirus, belonging to the family Paramyxoviridae, being classified as biosafety level 4 pathogens, due to their high capacity to cause lethal disease in humans associated with their unique genetic constitution, lack of specific therapy and prophylaxis. Bats belonging to the genus Pteropus, found in regions that extend from the Western Pacific to the East coast of Africa, are natural reservoir of such viruses. Deforestation is one of the factors responsible for the bats’ leaving their ecological niches and their adaptation to farms and villages. New cases of HeV infection in horses continue to occur in Australia while NiV has been responsible for annual human outbreaks since 2001 in India and Bangladesh. NiV has several features that highlight its potential as a pandemic threat, including its ability to infect humans directly from natural reservoirs, as well as a limited capacity for transmission between humans. Despite this, little is currently known about the mechanisms by which bats harbor viruses capable of causing serious diseases in other terrestrial mammals. This review provides relevant information for understanding the epidemiology of these viruses, the pathogenesis in susceptible species, as well as the importance of these viruses in domestic species, especially in horses. Keywords: Hendra Virus. Nipah Virus. Bats. Horses. Pigs.

O uso do tramadol vem sendo comumente e amplamente realizado na Medicina Veterinária nos últimos anos, o qual vem sendo utilizado tanto para dor aguda quanto para dor crônica. O tramadol é um opioide atípico com baixa afinidade por receptores µ (mu), e com ação monoaminérgica através da inibição da recaptação de norepinefrina e liberação de serotonina. O principal metabólito que é resultante da metabolização do tramadol nas isoenzimas do complexo P450 no fígado, é chamado de O-dismetiltramadol (M1), sendo o principal metabólito responsável pelo efeito analgésico do tramadol. No entanto, esse metabólito é encontrado em uma menor concentração em cães, quando comparado a gatos e humanos. E isso levou a um certo questionamento quanto a eficácia analgésica do tramadol em cães. Ademais, vários estudos já foram realizados avaliando os efeitos do tramadol, tanto em questões farmacocinéticas e farmacodinâmicas quanto em questões de avaliação de doses, vias de administração, e eficácia analgésica. Portanto, estudos a respeito do uso do tramadol em cães são crescentes, sendo necessário que os médicos veterinários se mantenham atualizados, para que possam empregar o fármaco de maneira eficaz. Diante de tais fatos, o presente trabalho teve como objetivo realizar uma revisão de literatura científica dos principais estudos relacionados ao uso do tramadol em cães. Palavras-chave: Analgesia. O-desmetiltramadol. Opióide. Tramadol. Abstract The use of tramadol has been commonly and widely used in Veterinary Medicine in recent years, which has been used for both acute and chronic pain. Tramadol is an atypical opioid with low affinity for µ receptors, and with monoaminergic action through inhibition of norepinephrine reuptake and serotonin release. The main metabolite that results from the metabolization of tramadol in the isoenzymes of the P450 complex in the liver, is called O-dismethyltramadol (M1), being the main metabolite responsible for the analgesic effect of tramadol. However, this metabolite is found in a lower concentration in dogs, when compared to cats and humans. This led to some questioning as to the analgesic efficacy of tramadol in dogs. In addition, several studies have been carried out evaluating the effects of tramadol, both on pharmacokinetic and pharmacodynamic issues and on questions of dose assessment, routes of administration, and analgesic efficacy. Therefore, studies on the use of tramadol in dogs are growing, and it is necessary that veterinarians keep up to date, so that they can use the drug effectively. Given these facts, the present study aimed to conduct a scientific literature review of the main studies related to the use of tramadol in dogs. Keywords: Analgesia. O-desmethyltramadol. Opioid. Tramadol.

Canine morbillivirus, also known as canine distemper virus (CDV), induces a contagious multisystemic disease caused by an enveloped RNA virus belonging to the genus Morbillivirus within the family Paramyxoviridae. CDV replicates readily in epithelial, nerve and lymphoid tissues; it is excreted in urine, feces, saliva, oral and nasal discharge; and its major route of entry for infection is through the respiratory system. Although the virus was originally believed to infect domestic dogs, new studies have shown that it can also naturally or experimentally infect non‐domestic hosts. A recent blood test performed on a giant anteater (Myrmecophaga tridactyla) found Lentz inclusions in the animal's leucocytes. A rapid CDV test, an RT‐PCR assay and pathology findings confirmed this report of canine morbillivirus in this species, which corresponds to the second report of CDV infection in the order Pilosa, family Myrmecophagidae in central west Brazil., This article is the first report of canine morbillivirus infection in a Giant anteater (Myrmecophaga tridactyla).The disease was confirmed through samples collected from the animal after the visualization of Lents corpuscles inside leucocytes during blood count cells.

The objective was to evaluate the effectiveness of mineral supplementation with organic selenium (Se) on rabies humoral immune response in ewes. It was randomly selected 18 Texel ewes without antirabic vaccination in the last 12 months and divided into 2 groups: treated group (TG) supplemented with a mineral mixture additioned of organic selenium and immunized against rabies and a control group (CG) supplemented with mineral mixture without organic selenium addition and immunized. Determination of Se serum was obtained by the enzyme glutathione peroxidase (GSH-Px) activity in two moments, the title and the persistence of anti-rabies neutralizing antibodies in five times by the Rapid Fluorescent Focus Inhibition Test (RFFIT). The mean values found in D210 GSH-Px were higher in TG with 981.8 ± 105.2 U/g Hb and 628.2 ± 188.2 U/g Hb in the CG (p-value < 0.001). The mean values of titers of anti-rabies neutralizing antibodies 30 days after primary vaccination, demonstrated statistically similar titles in the GT and GC, 1,020 IU/ mL in GT and 1,885 IU/ mL CG. Oral mineral supplementation of organic selenium during 210 days did not enhance the humoral immune response and persistence of evidence of rabies neutralizing antibodies, however caused increased serum retention of selenium in ewes.

Os norovirus foram os primeiros agentes virais ligados a doencas gastrointestinais em humanos. Por muito tempo, foram considerados como causa secundaria de gastroenterite, apos o rotavirus. O desenvolvimento de tecnicas moleculares favoreceu dados mais claros sobre o impacto epidemiologico dos norovirus, que sao atualmente reconhecidos como principal causa de surtos de gastroenterite nao bacteriana esporadica em criancas e adultos. Em analise da diversidade genetica e recombinacao dos norovirus sabe-se que os mesmos estao presentes em varias especies animais, incluindo os caes, o que denota na possivel exposicao dos seres humanos a estas novas cepas recem-descobertas. Devido a intensa relacao social atualmente observada entre humanos e animais de estimacao, sugerem-se novas investigacoes sobre o potencial zoonotico deste norovirus. O metodo de pesquisa utilizado neste trabalho foi uma revisao de literatura, que teve como objetivo revisar toda literatura atual sobre a epidemiologia e relevância do cao como potencial reservatorio para este agente. Ainda sao poucos os relatos sobre a presenca do norovirus em caes, que podem apresentar um potencial de transferencia zoonotica, porem devido a sua maior proximidade com os seres humanos na atualidade, considera-se a importância de novos estudos para avaliar o papel do cao como reservatorio do norovirus, em especial no Brasil, pela inexistencia de relatos ate o presente momento. Palavras-chave: Caes. Gastroenterite nao Bacteriana. Risco Zoonotico. Abstract Noroviruses were the first viral agents linked to gastrointestinal diseases in humans. For a long time, they were considered as secondary cause of gastroenteritis after rotavirus. The development of molecular techniques has favored clearer data on the epidemiological impact of noroviruses, which are currently recognized as the main cause of outbreaks of sporadic non-bacterial gastroenteritis in children and adults. In analyzing the genetic diversity and recombination of noroviruses it is known that they are present in several animal species, including dogs, which denotes the possible exposure of humans to these newly discovered strains. Due to the intense social relationship currently observed between humans and pets, further research on the zoonotic potential of this norovirus is suggested. The research method used is a review of the literature, which aimed to review all current literature on the epidemiology and relevance of the dog as potential reservoir for this agent. There are still few reports on the presence of norovirus in dogs, which may present a potential for zoonotic transfer, but due to its greater proximity to humans nowadays, the importance of new studies to evaluate the role of the dog as norovirus reservoir, especially in Brazil, due to the lack of reports to date. Keywords : Dog. Nonbacterial Gastroenteritis. Zoonotic Potential.

Feline Morbillivirus (FeMV) was first detected in 2012 in domestic cats from Hong Kong and was found to be associated with tubulointerstitial nephritis and chronic kidney disease. In subsequent studies in other countries, FeMV was detected in asymptomatic cats. However, it is not clear whether FeMV plays a role as a pathogen in the kidney diseases of cats, and other epidemiological data are still unknown. To date, studies have reported the presence of FeMV exclusively in domestic cats. This study is the first molecular detection of the FeMV RNA associated with pathological and immunohistochemical findings in a synanthropic marsupial, the white-eared opossum (Didelphis albiventris), inhabiting peri-urban areas of north-central Parana, Southern Brazil. Molecular techniques identified the viral RNA in the lungs and kidneys. Histopathologic evaluation of these tissues revealed interstitial pneumonia in the lungs with lymphocytic nephritis and tubular necrosis in the kidneys. Immunohistochemistry assays detected positive intralesional immunoreactivity to N protein of FeMV within the lungs and kidneys. A FeMV opossum strain was isolated in Crandell Rees feline kidney lineage cells, resulting in syncytia formation and cell death. Therefore, these results support the ability of FeMV to infect other mammal species and reinforce the possibility of the opossum to be a disseminator of this virus among domestic and wild animals.

Made available in DSpace on 2021-06-25T10:18:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Feline morbillivirus was discovered in 2012 in cats from Hong Kong, and it was initially found to be associated with chronic kidney disease. Although subsequent molecular surveys showed a common occurrence in cat populations from distinct countries, there were controversial results regarding the relationship between viral shedding through urine and reduced kidney function. In this study, 276 domestic cats of diverse origins from Western Brazil had their urine evaluated for the presence of paramyxoviral RNA by reverse transcription seminested PCR and direct sequencing. Additionally, a selected Brazilian feline morbillivirus strain was isolated in Crandell Rees feline kidney cells, and a nearly complete genome sequence was obtained. To assess the kidney function of all cats, serum biochemistry screening and standard urinalysis were performed. Our results revealed a relatively high paramyxovirus-positive rate (34.7%) in the evaluated cats although there was not a statistical association between the shedding of viral RNA through urine and kidney disease. Direct sequencing of partial fragments of the L gene demonstrated high genetic diversity among strains detected in cats in this study, since both feline morbillivirus RNA and feline paramyxovirus RNA were frequently shed in urine. Phylogenetic reconstruction based on partial amino acid sequences of the L gene showed that Brazilian feline paramyxovirus strains were genetically diverse since they grouped into two distinct subclusters; one subcluster contained three strains identified in Germany, while the second contained Japanese strain 163, which was recently classified in the Jeilongvirus genus of the Paramyxoviridae family. In contrast, the Brazilian feline morbillivirus strain FeMV/BR_Boni, herein characterized by nearly complete genome sequencing, was classified in the Morbillivirus genus with other strains previously identified as genotype 1. In conclusion, urinary excretion of diverse paramyxoviral RNA is frequent in cats of different origins from Western Brazil, but viral infection is not related to altered kidney function. Laboratory of Veterinary Microbiology Universidade de Cuiabá (UNIC) Laboratory of Animal Virology Department of Veterinary Preventive Medicine Universidade Estadual de Londrina (UEL) Institute of Biotechnology (IBTEC) Universidade Estadual Paulista (UNESP) Department of Small Animal Medicine Veterinary Teaching Hospital Universidade de Cuiabá Veterinary Clinical Pathology Universidade de Cuiabá Institute of Biotechnology (IBTEC) Universidade Estadual Paulista (UNESP)

Bluetongue (BT) is an infectious and non-contagious disease caused by bluetongue virus (BTV) belonging to the genus Orbivirus. It is transmitted by a hematophagous vector, Culicoides sp., to ruminants, particularly to sheep, which are most susceptible to this disease. The main serological tests are agar gel immunodiffusion (AGID), which is recommended by the World Organization for Animal Health (OIE), and the competitive enzyme-linked immunosorbent assay (cELISA), which has the advantage of no cross-reaction with other orbiviruses. The aim was to compare the results of these two tests by conducting them on sera collected from sheep in the state of Paraná, Brazil. From March to October 2017, serum samples were collected from 270 sheep from 10 farms in six mesoregions of Paraná. The samples were subjected to AGID and cELISA to detect antibodies against BTV. Based on the test results, we classified the sheep as low, moderate, and high occurrence. The results demonstrated that 64.81% (175/270) of the sheep were seropositive through the cELISA test, showing a high occurrence, and 41.11% (111/270) were seropositive through the AGID test, indicating a moderate occurrence. The concordance between the tests was moderate (0.51) as determined by the Kappa coefficient. Among the studied farms, 90% (9/10) presented at least one seropositive sheep, and the number of animals tested positive by the cELISA test was higher than those by the AGID test. Favorable climate, which favors the presence and multiplication of the culicoid vector and the occurrence of infection, was the biggest predominant factor responsible for the obtained results. The low occurrence in farms with milder climate suggest that the presence of antibodies also occurs due to the low pathogenicity of circulating serotypes in the different mesoregions studied. It is concluded that BTV infection is present in the sheep herds in Paraná, and the occurrence was moderate detected by AGID test and high detected by cELISA test. A língua azul (LA) é uma enfermidade infecciosa e não contagiosa causada por um vírus (VLA) do gênero Orbivirus, transmitida por vetores hematófagos Culicoides sp., aos ruminantes sobretudo aos ovinos, espécie mais susceptível. Os principais testes sorológicos utilizados são a Imunodifusão em Gél de Ágar (IDGA), preconizada pela OIE, e o teste Imunoensaio Enzimático Competitivo (ELISAc), sendo que este tem como vantagem não ocorrer reação cruzada contra outros orbivírus. O objetivo do trabalho foi detectar a presença de anticorpos contra o VLA em ovinos no estado do Paraná através dos testes diagnósticos IDGA e ELISAc. Durante os meses de março a outubro de 2017, colheu-se sangue de 270 ovinos, em 10 propriedades localizadas em seis mesorregiões paranaenses. As amostras foram submetidas aos testes de IDGA e ELISAc para detecção de anticorpos contra o VLA. Baseado nos resultados classificaram-se os rebanhos como baixa, moderada ou elevada ocorrência. Os resultados demonstraram elevada ocorrência através do teste de ELISAc, que apresentou 64,81% (175/270) de ovinos positivos, e moderada ocorrência através do IDGA, com 41,11% (111/270) de ovinos soropositivos. A concordância obtida entre os testes foi moderada (0,51) através do coeficiente Kappa. O número de ovinos reagentes no exame de ELISAc foi maior que o teste de IDGA em todas as propriedades positivas, demonstrando ser superior. O clima propício foi um dos fatores favoráveis para as ocorrências observadas, pois favorece a presença e multiplicação do vetor Culicoide e a ocorrência da infecção. A baixa ocorrência nas propriedades com clima mais ameno sugere que a presença de anticorpos provavelmente ocorra também pela baixa patogenicidade dos sorotipos circulantes nas diferentes mesorregiões estudadas. Conclui-se que há infecção de VLA no rebanho ovino paranaense, e a detecção de anticorpos para o VLA foi moderada, através do teste de IDGA, e elevada, através do teste ELISAc.

Canine distemper virus (CDV) is a highly contagious disease pathogen which causes disease in the domestic dog and species classified in the Canidae, Procyonidae, Mustelidae, Hyaenidae, Ursidae, Viveridae, Felidae, Tayassuidae, and Cercopithecidae families. A combined strategy that involved the direct sequencing of amplicons from genes coding for nucleocapsid, large polymerase, and hemagglutinin proteins of CDV, as well as the pathological findings and the immunohistochemical detection of viral nucleocapsid protein in diverse tissues, confirmed the participation of CDV in the development of a neurological disease in a southern tamandua ( Tamandua tetradactyla ) from Midwestern Brazil. Phylogenetic analysis based on the hemagglutinin gene sequences revealed that the strain from this study grouped with isolates from the Europe 1/South America 1 lineage. The specific polymorphisms at the SLAM receptor-binding site of the hemagglutinin gene, previously linked to disease emergence in novel hosts, were not detected in this genome. These findings represent the first description of CDV-induced infection in the Tamandua tetradactyla and extend the distribution of this infection to include members of the family Myrmecophagidae, order Pilosa.

Papillomaviruses (PVs) are complex viruses which infect the skin or mucosae of a broad range of amniotes worldwide. They cause benign or malignant lesions depending on environmental factors, virus oncogenicity and the location of infection. Bovine papillomaviruses (BPVs) are the second most studied PVs beyond human PVs. In the past few years, genetic characterization of animal PVs has increased due to the availability of new techniques, which simplified the sequencing of entire genomes. Therefore, this review aims to provide an update of the current epidemiology, classification and genome features of ruminant PVs (mainly BPVs) affecting animals worldwide. The review also aimed to clarify the key differences between the high-risk Delta papillomaviruses and the seemingly low-risk Xi, Epsilon, Dyoxi and Dyokappapillomavirus as well as the recently described PVs BPV18, 19, 21 and PpuPV1 that belongs to an unclassified genus.

Currently, bovine papillomavirus types are divided into five genera, namely, Deltapapillomavirus, Epsilonpapillomavirus, Xipapillomavirus, Dyoxipapillomavirus, and Dyokappapapillomavirus. In the recent decades, the characterization of numerous putative and novel bovine papillomavirus types from cattle in several geographic regions, has revealed the occurrence of a high viral diversity. In this study, we describe the identification and characterization of a putative new bovine papillomavirus type within species Xipapillomavirus 1 of Xipapillomavirus genus. The detection of the viral types identified in the skin warts was obtained by polymerase chain reaction assays targeting the L1 gene, followed by direct sequencing of the generated amplicons. The partial L1 sequences revealed that bovine papillomavirus types 6, 10, and 11, the putative new bovine papillomavirus type designated BPV/CHI-SW2, and an unreported putative new bovine papillomavirus type (named BPV/BR-UEL08) were associated with cutaneous papillomatosis in the cows from the dairy herd investigated. Phylogenetic reconstruction based on the L1 gene revealed that the BPV/BR-UEL08 isolate clustered with other bovine papillomaviruses classified in the Xipapillomavirus genus, being closely related to representatives of the species Xipapillomavirus 1. Investigations focusing on the molecular epidemiology of bovine papillomaviruses related to clinical outcomes in cattle are of fundamental importance to determine the actual genetic diversity and prevalent viral types to be included in vaccines for cattle.

The present review aims to show the main aspects related to bluetongue virus (BTV) infection in sheep. The bluetongue (BT) is a viral, infectious, and non-contagious disease caused by a virus (BTV) of the Orbivirus genus, transmited by a hematophagous vector of the Culicoides genus, to domestic and wild ruminants, mainly to sheep, the most susceptible species. It is caused by the association of endemic with climate conditions, with high temperatures and humidity. Economic loss is directly linked to death, abortion, weight loss, loss of milk, and meat production, and, indirectly, to the restriction on the export of animals and their by-products. The study concludes that the BTV is worldwidely spread, and probably persists due to the warm and humid climate that leads to the proliferation of Culicoides sp., being necessary to adopt measures that reduce the risk factors associated to the BTV infection. RESUMO A presente revisão objetivou apresentar os principais aspectos relacionados à infecção causada pelo vírus da língua azul em ovinos. A língua azul é uma doença viral, infecciosa e não contagiosa, causada por um vírus (BTV) do gênero Orbivírus, transmitida por meio de vetores hematófagos do gênero Culicoides a ruminantes domésticos e selvagens, principalmente aos ovinos, a espécie mais susceptível. A infecção ocorre de forma endêmica, associada a condições climáticas com elevada temperatura e umidade. As perdas econômicas estão ligadas diretamente à morte, ao abortamento, à perda de peso, à perda na produção de leite e carne, e, indiretamente, devido à restrição na exportação de animais e seus subprodutos. O estudo conclui que a língua azul está disseminada mundialmente e persiste, provavelmente, devido ao clima quente e úmido que propicia a proliferação de Culicoides sp., sendo necessário adotar medidas que diminuam os fatores de risco associados à infecção pelo vírus.

The objective of this study was to determine the gamma-glutamyl transferase (GGT) levels and its relationship with diarrhea and passive transfer of immunity in Holstein calves within 24 hours and 30 days of life from Leopolis municipality, the north Parana region. Colostrum is rich in immunoglobulins and vital for immunity to newborn calves, since bovine placenta does not allow the passage of immunoglobulin to the fetus. Calves undergo various challenges that can lead to disease and death in the first month of life, including diarrhea. Diarrhea has a multifactorial etiology, and the passive immunity transferred through ingestion of colostrum is able to protect the calf against many of these etiologic agents. GGT measurements indirectly infer the amount of immunoglobulin ingested by the calf. Higher serum GGT levels (381.72 IU / L) were found at 24 hours, and a significant reduction was observed at 30 days (66.22 IU / L). When the presence or absence of diarrhea was associated with GGT levels above and below 200 IU / L, no statistical significance (P> 0.05) was observed, since 80% of animals with diarrhea had serum GGT levels higher than 200 IU / L. Under the conditions of this study, there was no relationship between the GGT concentration and the occurrence of diarrhea, and no mortality was observed despite some animals presented diarrhea.

The 30th meeting of the Brazilian Society for Virology (SBV) was held, for the first time in its 30 years of existence, in Cuiabá, the capital of Mato Grosso State, Central Western Brazil, a tropical region between the three richest biomes in the world: Amazon Florest, Cerrado and Pantanal. In recent years, the field of virology has been built in the State. The aim of this report is to support participants and virologists to receive the most up-to-date information about the meeting, which occurred from 16 to 19 October 2019. National and international speakers gave SBV the opportunity to learn about their experience on their virology fields, sharing recent scientific findings, compiling conferences, round table presentations and work presentations in oral and poster sessions. The meeting held over 300 attendants, who were also involved on oral and poster presentations, showing a great variety of recent unpublished studies on environmental, basic, animal, human, plant and invertebrate virology. In addition, SBV offered the Helio Gelli Pereira award for the best research studies in each field presented during the meeting. The 30th meeting of SBV was very productive and has also encouraged scientific partnership and collaboration among virologists worldwide.

Resumo:Sarcoides são tumores fibroblásticos, considerados os tumores de pele mais comuns em pele de equinos e que raramente apresentam regressão espontânea. Papilomavírus bovino (BPV) tipos 1 e 2 são relacionados com a patogenia do sarcoide e, provavelmente, o BPV tipo 13 (BPV13), recentemente descrito, também pode estar associado com a formação dessa lesão. Neste estudo, 20 amostras de lesões cutâneas, sendo 12 constituídas por tecidos frescos e 8 amostras de tecido fixado em formalina e embebido em parafina, provenientes de 15 cavalos foram utilizadas para a identificação do DNA de BPV. A análise histopatológica (HE) confirmou todas as lesões como sarcoide. Para a amplificação do DNA de papilomavírus (PV) foram realizadas três reações de PCR. Como triagem, os primers IFNR2/IDNT2 foram utilizados para amplificar um fragmento da ORF L1 do PV. O segundo par de primersutilizado é complementar a sequência dos genes E5 e L2 de BPVs 1, 2 e 13. O terceiro par de primers(FAP59/FAP64) utilizado tem o gene L1 como alvo. A primeira e a segunda PCRs permitiram amplificar produtos em todas as amostras avaliadas. Entretanto, na terceira reação, na qual foram utilizados os primers FAP, foi possível amplificar produtos com tamanho molecular esperado somente nas amostras constituídas por tecidos frescos. O sequenciamento de nucleotídeos e as análises filogenéticas realizadas nos fragmentos E5L2 resultaram na identificação de BPV1, 2 e 13 em 14 (70%), 2 (10%) e em 4 (20%) amostras de sarcoides, respectivamente. As amostras de sarcoides de um dos animais continha somente o DNA de BPV1. Entretanto, nas amostras provenientes do segundo cavalo foi possível identificar o DNA de três tipos de Deltapapillomavirus bovino (BPV1, 2 e 13) em lesões distintas. Este estudo ratifica a presença do DNA de BPV1, 2 e 13 em lesões de sarcoides em equinos, além de identificar três tipos de BPVs em um mesmo animal e descrever pela primeira vez no Brasil a presença de BPV1 e 2 nesse tipo de lesão. Abstract:Sarcoids are fibroblastic lesions, which are considered as the most common skin tumors of horses; spontaneous regression rarely occurs. The bovine papillomavirus (BPV) types 1 and 2 may be involved in the pathogenesis of sarcoids, and probably the recently described BPV type (BPV13) might be associated with the pathogenesis of this lesion. This study characterized the DNA of BPVs in sarcoids from 15 horses from Brazil by analyzing 20 cutaneous lesions (12 recently collected; 8 from formalin-fixed paraffin-embedded (FFPE) tissues). Histopathology confirmed the proliferative lesions as sarcoids. Three PCRs were performed to amplify papillomavirus (PV) DNA. For screening, the primers IFNR2/IDNT2 were used to amplify a fragment of the PV L1 ORF. The second primer set was complementary to a common sequence of the E5L2 genomic region of BPV1, 2, and 13. The third primer pair (FAP59/FAP64) targeted a fragment of the PVs L1 ORF. The screening and E5L2 PCRs yielded amplicons in all samples evaluated. The FAP amplicons identified BPV1, 2, and 13 only from fresh tissue samples. The phylogenetic analyses of E5L2 resulted in the identification of BPV1, 2, and 13 in 14 (70%), 2 (10%), and 4 (20%) sarcoids, respectively. Two horses demonstrated multiple lesions: the sarcoids of one of these contained only BPV1 DNA and those of the other contained three types of bovine Deltapapillomavirus (BPV1, 2, and 13). This study confirmed the presence of BPV1, 2, and 13 DNA in equine sarcoids. Moreover, these findings represent the first description of three types of BPV diagnosed in the same horse, as well as the first confirmation of BPV1 and 2 in horses from Brazil.

Bacteria classified in Mycoplasma (M. bovis and M. bovigenitalium) and Ureaplasma (U. diversum) genera are associated with granular vulvovaginitis that affect heifers and cows at reproductive age. The traditional means for detection and speciation of mollicutes from clinical samples have been culture and serology. However, challenges experienced with these laboratory methods have hampered assessment of their impact in pathogenesis and epidemiology in cattle worldwide. The aim of this study was to develop a PCR strategy to detect and primarily discriminate between the main species of mollicutes associated with reproductive disorders of cattle in uncultured clinical samples. In order to amplify the 16S-23S rRNA internal transcribed spacer region of the genome, a consensual and species-specific nested-PCR assay was developed to identify and discriminate between main species of mollicutes. In addition, 31 vaginal swab samples from dairy and beef affected cows were investigated. This nested-PCR strategy was successfully employed in the diagnosis of single and mixed mollicute infections of diseased cows from cattle herds from Brazil. The developed system enabled the rapid and unambiguous identification of the main mollicute species known to be associated with this cattle reproductive disorder through differential amplification of partial fragments of the ITS region of mollicute genomes. The development of rapid and sensitive tools for mollicute detection and discrimination without the need for previous cultures or sequencing of PCR products is a high priority for accurate diagnosis in animal health. Therefore, the PCR strategy described herein may be helpful for diagnosis of this class of bacteria in genital swabs submitted to veterinary diagnostic laboratories, not demanding expertise in mycoplasma culture and identification.

The aim of this study was to investigate the prevalence of Brucella abortus infection in employees of two slaughterhouses (A and B) in the metropolitan area of Cuiabá in the state of Mato Grosso and the risk factors associated with its transmission. We collected 134 blood samples from workers in all sections of the slaughterhouses to evaluate brucellosis, and we administered a questionnaire to obtain information about potential risk factors associated with its transmission. We also collected blood from 468 cows before slaughter on the same day of the blood collection from the slaughterhouse workers to determine bovine seropositivity. Six (4.5%) out of the 134 workers examined were positive for B. abortus infection, and all worked as slaughterers. We found a significant association between the prevalence of brucellosis and slaughterers compared to other occupations at the slaughterhouse. It is noteworthy that four seropositive workers had worked as a slaughterer in other establishments, they did not use personal protective equipment (PPE) and they had consumed dairy products without heat treatment. In assessing the prevalence of brucellosis in slaughtered animals in both slaughterhouses, there was a higher number of seropositive cows at slaughterhouse B than slaughterhouse A: of the 468 cows tested in both slaughterhouses, 35 (7.5%) were positive for B. abortus infection, as determined by the 2-mercaptoethanol test (2-ME). Slaughterhouse workers are susceptible to infection by B. abortus, and it was noted that in both slaughterhouses, all of the seropositive workers were slaughterers. Cows that were positive for B. abortus were slaughtered in both slaughterhouses without proper sanitary procedures and necessary information for slaughter. Not using personal protective equipment and the slaughter of animals that are seropositive for brucellosis are risk factors for the transmission of the disease to slaughterhouse workers. O objetivo deste trabalho foi estudar a prevalência da infecção por Brucella abortus em trabalhadores de duas indústrias frigoríficas (A e B) na região metropolitana de Cuiabá, Mato Grosso, e os fatores de risco associados a sua transmissão. Foram colhidas 134 amostras de sangue dos trabalhadores, em todos os setores das indústrias, para exame de brucelose e aplicado questionário para levantamento de informações de possíveis fatores de risco associados à transmissão da doença. Realizou-se também, a colheita de sangue em 468 fêmeas bovinas antes do abate, no mesmo dia da colheita de sangue dos funcionários das indústrias pesquisadas, para identificação da soropositividade bovina. Dos 134 trabalhadores examinados, seis (4,5%) foram reagentes para a infecção por B. abortus e todos trabalhavam na atividade de magarefe. Obteve-se associação significativa da prevalência de brucelose em magarefes, em relação às demais atividades desenvolvidas na indústria. Quatro trabalhadores soropositivos já haviam trabalhado na função de magarefe em outros estabelecimentos, não utilizavam equipamento de proteção individual (EPI) e já haviam consumido produtos lácteos sem tratamento térmico. Ao avaliar a prevalência de brucelose nos animais abatidos nas duas indústrias frigoríficas, encontrou-se maior número de fêmeas soropositivas na indústria B do que na indústria A onde, das 468 fêmeas com sangue colhidos nas duas indústrias, 35 (7,5%) reagiram positivamente a infecção por B. abortus pela prova de 2-mercaptoetanol (2-ME). Trabalhadores das indústrias frigoríficas estão susceptíveis a contaminação por B. abortus, sendo observado nas indústrias que as pessoas soropositivas desenvolviam atividades de magarefe. Fêmeas bovinas soropositivas a B. abortus foram abatidas nas indústrias frigoríficas sem a devida informação e procedimentos sanitários necessários ao abate. As atividades de magarefe, a não utilização de EPI e o abate de animais soropositivos a brucelose são fatores de risco à transmissão da enfermidade para os trabalhadores da indústria frigorífica.

Bovine viral diarrhoea virus (BVDV) is an important pathogen of cattle that occurs worldwide with substantial economic impact on beef and dairy industries. The aim of this study was to describe the diversity of BVDV subgenotypes in persistently infected (PI) animals identified in a highly productive, regularly vaccinated, dairy cattle herd presenting with reproductive failure. Serum samples were collected from all animals within the herd (n = 692) and used to detect the presence of BVDV RNA. Using reverse transcription polymerase chain reaction assay, 29 cows were identified as transiently infected, three animals (two cows and one calf) as persistently infected, and one calf as putative BVDV PI animal. The sequences of 5'UTR and/or N(pro) gene of BVDV used in phylogenetic analyses revealed that the three PI animals were infected by three different BVDV subgenotypes (BVDV-1a, BVDV-1b, and BVDV-1d). These results demonstrated that in an open dairy cattle herd, regular vaccination against BVDV by itself is not able to prevent viral circulation in the herd. Furthermore, depending on the frequency of the acquisition of heifers and/or cows for replacement, several BVDV subgenotypes may co-exist simultaneously in the same herd.

A infecção pelo vírus da diarreia viral bovina (BVDV) foi avaliada em um rebanho bovino leiteiro de alta produção com histórico de problemas reprodutivos e de vacinação regular contra o BVDV. A identificação do vírus foi realizada por RT-PCR em soro sanguíneo e o perfil sorológico por vírus-neutralização. Inicialmente, 100% (n=692) dos animais do rebanho foram avaliados com relação à presença de infecção ativa pelo BVDV por meio da RT-PCR. Quatro meses após, todos os animais positivos (n=29) na primeira avaliação foram avaliados novamente pela RT-PCR, assim como todos os animais que nasceram (n=72) e os que apresentaram problemas reprodutivos (n=36) no intervalo entre a primeira e a segunda colheita de sangue. Os resultados finais do estudo possibilitaram identificar 27 animais transitoriamente infectados e três animais persistentemente infectados (PI). A sorologia, realizada apenas nos animais positivos na primeira avaliação pela RT-PCR e nas vacas que apresentaram problemas reprodutivos entre a primeira e a segunda RT-PCR, demonstrou grande flutuação nos títulos de anticorpos neutralizantes, além de soroconversão na maioria dos animais. Foram identificados aumentos nos títulos de anticorpos neutralizantes que variaram entre 3 e 8 log2, indicando infecção ativa no rebanho. A circulação viral no rebanho avaliado foi responsável pela expressão de sinais clínicos da esfera reprodutiva em animais com baixo título de anticorpos e consequente falha na proteção fetal. Os resultados demonstram que o controle da infecção pelo BVDV apenas por meio da vacinação regular em rebanhos com animais PI pode não ser eficaz na profilaxia dessa virose. The profile of bovine viral diarrhea virus (BVDV) infection was studies in a high production dairy herd selected based on a history of reproductive failures and regular vaccination against BVDV. Virus identification was performed by RT-PCR and serological profile was determined by virus-neutralization (VN). Initially, 100% (n=692) of the animals in the herd were monitored for identification of an active infection by RT-PCR in sera. Four months later, all positive animals (n=29) were retested by RT-PCR, along with newly born animals (n=72), or those that had reproductive failures (n=36) in the interval. The RT-PCR assay identified 27 transiently infected animals and three persistently infected (PI). Serology performed only in positive animals in the first RT-PCR and in cows with reproductive failures between the first and second RT-PCR analysis, showed large variation VN antibody titers and seroconversion in most animals. Increases in VN titers were demonstrated, with variation between 3 and 8 log2, indicating virus circulation within the herd. Virus circulation in the vaccinated herd evaluated in this study was likely responsible for reproductive failures observed in cows with low VN titers and for fetal infections. These results demonstrate that control of BVDV infection by regular vaccination in dairy cattle herds with PI animals represents a great challenge for the prophylaxis of this infection.

This report describes the complete genomic sequence and taxonomic position of BPV type 13. The BPV13 genome was amplified using the multiply primed rolling-circle amplification technique and long-template PCR employing two specific primers. The two long PCR fragments obtained were cloned and sequenced via primer walking. The complete genomic sequence of the BPV13 contains 7961 bp encoding eight proteins, E1, E2, E4, E5, E6, E7, L1, and L2. Similarly to the E5 gene in BPVs 1 and 2, the putative BPV13 E5 ORF encodes a small transforming protein that contains a hydrophobic transmembrane domain. Meanwhile, the retinoblastoma tumor suppressor-binding domain is absent in the putative BPV13 E7 protein. The presence of these two specific molecular features has been recognized as a distinct marker for the development of fibropapilloma in artiodactyl PV-induced lesions. The phylogenetic analysis demonstrated that BPV13 is a new member of the Deltapapillomavirus genus, to be classified as the third representative of the Delta 4 species. The characterization of the genomic sequence of this novel PV will aid in the interpretation of the pathologies described to be related to this virus and provide support for the development of diagnostic tools for epidemiological surveillance of BPV13 in its potential natural hosts.

Feline morbillivirus was first identified in healthy and diseased stray cats captured in Hong Kong. Recently, it was demonstrated that the virus circulates within cat populations in Japan, Italy, Germany, and the USA. Importantly, an association between feline morbillivirus infection and chronic kidney disease was suggested by histological analysis of kidney tissue of infected cats. The aim of this study was to verify the presence and examine the genetic diversity of feline morbilliviruses associated with infections of domestic cats in Brazil. Seventeen cats without clinical manifestations of urinary tract diseases from a multi-cat household and 35 random client-owned cats admitted to the Teaching Veterinary Hospital for a variety of reasons were evaluated for paramyxoviral infection and the presence of uropathy. A fragment of the paramyxoviral L gene was amplified from urine samples using a reverse transcription semi-nested PCR assay. For the first time, we detected a feline morbillivirus strain that was genetically related to viral strains previously characterized in Japan in urine samples from cats in South America, in Brazil. This together with the recent description of feline morbillivirus identification within cat populations in the USA, suggests a possible widespread distribution of this viral agent on the American continent. Our data demonstrated feline morbillivirus RNA shedding mostly in the urine of cats without clinical, laboratorial, or ultrasonographic signs of urinary tract diseases. In contrast to previously published findings that associated feline morbillivirus infection with chronic kidney disease, we did not observe a clear relationship between feline morbillivirus RNA shedding in urine and kidney disease in the cats evaluated.

Bovine herpesvirus 5 is a DNA virus that has been associated with meningoencephalitis in young cattle. While its clinical diagnosis is obscured by other major diseases that also produce similar neurological disease in cattle, the use of conventional virological techniques is hampered by the establishment of a lifelong latent infection in the host and the difficulty in differentiating BoHV-1 and BoHV-5. The aim of the current report is to describe the clinical and epidemiological aspects observed in a natural outbreak of BoHV-5 meningoencephalitis in a dairy cattle herd from Brazil. In the outbreak, the affected animals consisted of nine calves, which presented three possible forms of the neurological disease, subjectively classified as peracute, acute, and subacute/chronic. In contrast to conventional herpetic meningoencephalitis, characterized mainly by progressive multifocal brain dysfunctions, BoHV-5 infection resulted in focal non-progressive caudal brainstem dysfunction (pontomedullary syndrome) in an animal presented with subacute/chronic BoHV-5 meningoencephalitis. The evaluation of CNS tissue of affected calves through both histological examination and multiplex-PCR was able to confirm BoHV-5 infection. Additionally, the analysis of CSF samples through PCR allowed ante-mortem BoHV-5 diagnosis during the outbreak, which enabled the implementation of several measures of control for the disease.O herpesvírus bovino 5 é um vírus DNA que tem sido associado a casos de meningoencefalite em bovinos jovens. Enquanto o diagnóstico clínico da doença é dificultado pelo fato de outras enfermidades importantes também determinarem quadro neurológico em bovinos, a utilização de técnicas virológicas convencionais no diagnóstico laboratorial tem sido inviabilizada pela latência viral que ocorre no hospedeiro e pela dificuldade em se diferenciar o BoHV-1 do BoHV-5. O objetivo deste relato é o de descrever alguns aspectos clínicos e epidemiológicos observados em um surto de meningoencefalite determinado pelo BoHV-5 em um rebanho bovino leiteiro do estado do Paraná, Brasil. Neste surto, nove bezerros afetados apresentaram três formas distintas da doença, subjetivamente classificadas como hiperaguda, aguda e subaguda/crônica. Diferentemente da meningoencefalite herpética convencional, caracterizada principalmente por disfunções cerebrais multifocais progressivas, a infecção pelo BoHV-5 resultou em disfunção focal não-progressiva do tronco encefálico (síndrome pontobulbar) em um animal apresentando a meningoencefalite subaguda/crônica por BoHV-5. As avaliações de tecido do sistema nervoso de bezerros afetados, por meio do exame histológico e da técnica de multiplex-PCR, confirmaram a infecção pelo BoHV-5. Adicionalmente, a análise de amostras de líquor por meio da PCR permitiu o diagnóstico ante-mortem da infecção pelo BoHV-5, fato que possibilitou a implementação de várias medidas de controle para a doença durante o surto.

The amplification by degenerate primers FAP59/FAP64 and sequencing allowed the detection of 15 putative new BPV types in cutaneous warts as well as in healthy skin. Four of these isolates were recently recognized as new BPV types (BPV-7, -8, -9, and -10) after determination of their complete genome sequences. In Brazil, investigations involving the definition of BPV types present in skin warts are still rare. The aim of the current study was to identify the BPV types associated with cutaneous papillomatosis observed in Brazilian cattle herds. Twenty-two cutaneous papilloma specimens were submitted to PCR assay employing the FAP primer pair. All PCR products with approximately 480 bp were submitted to direct sequencing. Cloning was performed for the amplicons which prior analysis revealed as putative new BPV types. From 16 cutaneous lesions, BPV-1, -2, and -6 were identified in two, six, and eight papilloma specimens, respectively. In addition, four putative new BPV types were identified in other six skin warts, and then designated as BPV/BR-UEL2 to -5. The detection of the BPV-1, -2, and -6 types in skin wart specimens supports the existence of these BPV types throughout the Brazilian cattle herd. In addition, the identification of four putative new BPV types is the first report of the presence of different BPV types in the American continent.

When first described in 1946, bovine viral diarrhea (BVD) was characterized as an acute transmissible disease associated with severe leucopenia, high fever, depression, diarrhea, gastrointestinal erosions, and hemorrhages. Recently the severe acute form has been related only to some hypervirulent BVDV-2 strains. This article reports the detection of BVDV-1b associated with an acute and fatal outbreak of BVD in a Brazilian beef cattle herd. Depression, anorexia, watery diarrhea, sialorrhea, and weakness were observed in six steers. One of these animals was evaluated for laboratorial, clinical, and pathological alterations. Laboratory findings were non-specific; clinically, the animal was weak, with dehydration and erosive oral lesions. Pathological alterations were predominant at the tongue, esophagus, and rumen. A RT-PCR assay using primers to partially amplify the 5' untranslated region (5'UTR) of the BVDV genome was performed and identified BVDV in all clinical samples analyzed. Phylogenetic analysis of BVDV derived from lymph node revealed that this strain was clustered within the BVDV subtype 1b. This differentiating was only possible to be performed by molecular characterization since both clinical presentation and pathologic findings were similar to BVDV-2 infection.

Bovine herpesvirus 5 (BoHV-5) is an important cause of meningoencephalitis in young and adult cattle. The multiple etiology of neurological disturbances in cattle makes the quick and conclusive diagnosis of BoHV-5 infection important for animal and public health, mainly because of herbivore rabies that is endemic in Brazilian cattle herds. The objective of this retrospective study was to use a multiplex-polymerase chain reaction (multiplex-PCR) for BoHV-5 and BoHV-1 glycoprotein C gene detection from stored central nervous system (CNS) tissue fragments of cattle with neurological clinical signs. Forty-seven frozen CNS samples of young and adult cattle from 31 herds in three Brazilian geographical regions (South, Southeast, and Center-west) were evaluated. Eighteen (38.3%) of these CNS samples were BoHV-positive by virus isolation in cell culture. By multiplex-PCR 30 (63.8%) CNS samples were BoHV-5 positive. All 18 positive samples by virus isolation were confirmed as BoHV-5 by the multiplex-PCR, that provided a increase of 25.5% (12/47) in the BoHV-5 diagnosis rate. BoHV-1 was not detected in any CNS sample. This retrospective study demonstrated the wide regional distribution of BoHV-5 infection in Brazilian cattle herds since positive results were obtained in CNS samples of cattle with neurological disease from Paraná, São Paulo, Minas Gerais, Mato Grosso, and Mato Grosso do Sul States.O herpesvírus bovino 5 (BoHV-5) é um importante agente etiológico de meningoencefalite em bovinos jovens e adultos. A etiologia múltipla dos distúrbios neurológicos em bovinos torna o diagnóstico conclusivo do BoHV-5 importante tanto em termos de sanidade animal quanto de saúde pública, principalmente pela característica endêmica da raiva dos herbívoros nos rebanhos bovinos brasileiros. O objetivo desse estudo retrospectivo foi utilizar a reação em cadeia da polimerase (multiplex-PCR) para a detecção do gene da glicoproteína C do BoHV-5 e do BoHV-1 em fragmentos estocados de sistema nervoso central (SNC) de bovinos com sinais clínicos neurológicos. Foram avaliadas 47 amostras congeladas de fragmentos de SNC de bovinos jovens e adultos pertencentes a 31 rebanhos de três regiões geográficas brasileiras (Sul, Sudeste e Centro-oeste). Por meio do isolamento viral em cultivo celular foi possível o isolamento do BoHV em 18 (38,3%) amostras. Pela técnica de multiplex-PCR 30 (63,8%) amostras de SNC foram positivas para o BoHV-5. Todas as 18 amostras positivas no isolamento viral foram confirmadas como BoHV-5 pela multiplex-PCR, proporcionando um incremento na taxa de diagnóstico do BoHV-5 de 25,5% (12/47). Em nenhuma das amostras avaliadas foi possível a identificação do BoHV-1 pela multiplex-PCR. Esse estudo retrospectivo demonstrou a ampla distribuição da infecção pelo BoHV-5 nos rebanhos bovinos brasileiros uma vez que resultados positivos foram obtidos em amostras de SNC colhidas de bovinos com doença neurológica, provenientes dos estados do Paraná, São Paulo, Minas Gerais, Mato Grosso e Mato Grosso do Sul.

Teat papillomatosis affects dairy cows worldwide. Milking can become difficult due to teat warts, and maintaining affected cows in the herds may diminish economic profit in the dairy industry. Currently, 13 bovine papillomavirus (BPV) types have been fully characterized, and numerous putative BPV types have been identified through partial L1 gene PCR. In order to identify the viral types present in warts on the udders of dairy cows, 40 teat lesions from 24 cows from 13 cattle farms in three States of Brazil were evaluated by PV L1 gene PCR. The warts that were evaluated contained sequences from BPVs 6-10, the putative BPV types BAPV9 and BAPV4, and two unreported putative papillomavirus (PV) types, named BPV/BR-UEL6 and BPV/BR-UEL7. In addition, mixed infections and coinfections were identified, since more than one lesion was observed on the udders of 13 cows. Phylogenetic analysis showed that BPV/BR-UEL6 is closely related to BPVs belonging to the genus Xipapillomavirus, while BPV/BR-UEL7 clustered with the previously reported strains Cervus timorensis and Pudu puda PVs, which represent a putative new PV type, and it was only distantly related to xi-, epsilon-, delta- and dyoxi-PVs. These results provide information that will assist in the understanding of the association of BPVs 6, 7, 8, 9, and 10, as well as putative BPV types BAPV4 and BAPV9, with mammary papillomatosis. This is the first characterization of putative novel PV types BPV/BR-UEL6 and BPV/BR-UEL7 in teat warts of dairy cows, highlighting the high genetic diversity of BPVs associated with teat papillomatosis.

Equine sarcoids are locally aggressive fibroblastic neoplasms considered to be the most common skin tumors of horses worldwide. Bovine papillomavirus types 1 and 2 have typically been associated with sarcoids in equids. Investigations aiming to identify papillomavirus strains, aside from bovine papillomaviruses 1 and 2, which might be associated with sarcoid lesions, have been lacking. The aim of this article is to report the identification of a third bovine papillomavirus type, bovine papillomavirus 13, associated with equine sarcoids. Six sarcoid lesions were collected from diverse anatomical sites on two horses from southern Brazil. To detect a broad spectrum of papillomavirus strains, eight degenerate primer pairs designed to detect conserved regions on the L1 and E1 genes were tested on the DNA samples. Direct sequencing was then performed on the obtained amplicons, and sequence identities were compared with sequences from all bovine papillomavirus types. The FAP59/FAP64, MY09/MY11, and AR-E1F2/AR-E1R4 sequences generated from the sarcoids were shown to present 99 to 100% identity with bovine papillomavirus 13, a new bovine papillomavirus type previously described in cattle. The results from this study suggest that there is a need to identify bovine papillomavirus type 13 and other papillomavirus strains that might be associated with sarcoids in diverse geographical areas; such investigations might establish the frequency of occurrence of this viral type in these common tumors of equids.

O papilomavírus (PV) pertence a um grupo muito diversificado de vírus que infecta grande variedade de espécies de vertebrados. O PV está relacionado com proliferações da pele e mucosa de seus hospedeiros. Em bovinos, o papilomavírus bovino (BPV) é o agente etiológico da papilomatose cutânea e dos cânceres da bexiga urinária e do trato gastrointestinal superior. Nos equinos, os sarcóides são neoplasias fibroblásticas locais com característica agressiva, reconhecidas como o tumor de pele mais comum em equinos de todo o mundo. Os sarcóides raramente regridem e, frequentemente, recorrem após a terapia. Pela identificação sucessiva do DNA do BPV em sarcóides, e a demonstração da expressão de diversos genes virais nestas lesões, o envolvimento direto do BPV na patogênese do sarcóide foi corroborado. Atualmente, doze tipos de BPV foram identificados e caracterizados em bovinos. Estes BPVs estão classificados nos gêneros Deltapapillomavirus (BPV1 e 2), Xipapillomavirus (BPV3, 4, 6, 9, 10, 11, e 12) e Epsilonpapillomavirus (BPV5 e 8), com a exceção do BPV7 que pertence a um gênero ainda indefinido. Além dos tipos de BPV caracterizados por meio da sequência completa do genoma viral, dezenas de prováveis novos tipos virais, identificados por meio de PCR com primers degenerados que amplificam fragmentos parciais do gene L1, têm sido descritos em rebanhos bovinos provenientes de regiões geográficas diversas. Recentemente, estudos realizados no Brasil revelaram notável diversidade entre os BPVs detectados em papilomas bovinos, sendo descritos quatro prováveis novos tipos virais que foram designados BPV/BR-UEL2 a BPV/BR-UEL5. O presente estudo descreve a sequência genômica completa e o posicionamento filogenético de um destes novos tipos de BPV (BPV/BR-UEL4) identificado em bovinos e tentativamente denominado de papilomavírus bovino tipo 13. Adicionalmente, o envolvimento deste novo tipo de BPV em sarcóides equinos também é relatado. Em cavalos, seis sarcóides que foram coletados cirurgicamente, a partir de várias regiões anatômicas de dois cavalos, foram avaliados com oito pares de primers degenerados. Estes primers foram desenhados para detectar regiões conservadas dos genes L1 e E1 dos PVs cutâneos. Os produtos de PCR obtidos foram sequenciados diretamente. Nos sarcóides avaliados, as quatro sequências geradas dos produtos de PCR apresentaram 100% de identidade com o BPV13. O BPV13 foi identificado em um papilomas localizado na orelha de bovino pertencente a um rebanho da região sul do Brasil. Para amplificar a sequência completa do genoma do BPV13, a amostra de DNA foi submetida à técnica de amplificação por círculo rolante (RCA). Para confirmar que o produto obtido era DNA de papilomavírus, PCR com primers degenerados foi realizada no produto RCA. Estes produtos de PCR foram sequenciados e, baseando-se nestas sequências parciais dos genes E1 e L1, dois pares de primers para PCR de fragmentos longos foram selecionados para a amplificação da maior parte do genoma do BPV13. A sequência completa dos fragmentos longos clonados foi determinada por sequenciamento contíguo, iniciando-se a partir do sítio para os primers universais no vetor de clonagem pCR4-TOPO. As falhas remanescentes nas sequências foram eliminadas por sequenciamento direto do produto RCA. A sequência genômica completa do BPV13 possue 7961 pb, com conteúdo GC de 45,1%, e oito ORFs, codificando para seis proteínas não-estruturais (E1, E2, E4, E5, E6 e E7) e duas proteínas estruturais (L1 e L2). A análise filogenética, realizada a partir do alinhamento de sequência de nucleotídeos concatenada dos genes E7, E1, E2, L2 e L1 do BPV13 e de outros PVs de ungulados, mostrou que o BPV13 é um membro do gênero Deltapapillomavirus, que já contém os BPVs 1 e 2. A sequência do gene L1 do BPV13 apresentou identidade de 85,5-88,2% com as mesmas sequências dos BPVs 1 e 2. A identificação do BPV13 nos sarcóides examinados sugere a possibilidade de que muitos PVs isolados de sarcóides, e que foram previamente diagnosticados por meio de PCR com primers específicos para o BPV1/2, podem ter sido classificados erroneamente como BPV2, especialmente devido ao fato do sequenciamento direto não ter sido realizado para muitas amostras. Assim como o relatado para os BPVs 1 e 2, é tentador sugerir que o BPV13 também é capaz de induzir fibropapilomas em seus hospedeiros naturais. O agrupamento do BPV13 no gênero Deltapapillomavirus, o qual reune PVs que determinam fibropapilomas em artiodactilos ruminantes, assim como os achados da presença do gene E5 e da perda do motivo de ligação do fator pRB na proteína E7, dão suporte a esta hipótese. Os PVs são espécie-específicos e a ocorrência de transmissão entre espécies é relatada como evento muito raro, somente ocorrendo entre espécies hospedeiras próximas, como a infecção de cavalos pelos BPVs 1 e 2. Assim como os outros representantes da espécie Delta4, este novo tipo viral foi identificado em associação com casos de sarcóide equino em rebanhos brasileiros. A caracterização de novos BPVs colabora para a compreensão da filogenia do PV e para a interpretação das enfermidades relacionadas a estes vírus. A obtenção de informações adicionais sobre a base molecular e epidemiologia dos BPVs caracterizados, assim como dos ainda não caracterizados, será necessária para a completa compreensão da patogenia das doenças associadas ao BPV. Papillomaviruses (PVs) represent a large and highly diverse group of pathogens that infect a wide variety of vertebrate species. PVs are linked to epithelial proliferations of the skin or the mucosa of their hosts. In cattle, the bovine papillomavirus (BPV) is thought to be the casual agent of cutaneous papillomatosis, cancer of the urinary bladder and upper gastrointestinal tract. Equine sarcoids are locally aggressive fibroblastic neoplasms, considered as the most common skin tumor in horses worldwide. These tumors rarely regress and very often recur after therapy. Through consistent identification of BPV DNA in sarcoids as well as the demonstration of expression of diverse viral genes, the direct involvement of BPV in the pathogenesis of sarcoid tumors was corroborated. Until now, twelve BPV types were identified and characterized from cattle. These BPVs are classified in the genera Deltapapillomavirus (BPV1 and 2), Xipapillomavirus (BPV3, 4, 6, 9, 10, 11, and 12) and Epsilonpapillomavirus (BPV5 and 8), with the exception of BPV7, which belongs to an undesignated PV genus. In addition to the fully sequenced BPV types, through PCR assay employing consensus or degenerate primers which amplify partial fragments of the L1, the presence of numerous putative new BPV types have been described in cattle herds from diverse geographical regions. Recently, an investigation using the strategy above mentioned revealed notable diversity among BPVs detected in papillomas from Brazilian cattle herds. The study identified four putative new BPV types designated as BPV/BR-UEL2 to BPV/BRUEL5. This study describes the complete genomic sequence and phylogenetic position of one of these novel BPV types (BPV/BR-UEL4) isolated from cattle: the bovine papillomavirus type 13. Additionally, it reports the involvement of this new BPV type in equine sarcoid lesions. In horses, six sarcoids which were individually and surgically collected from diverse anatomic locations of two horses were tested with eight degenerate primer pairs, designed to detect conserved regions on L1 and E1 genes of a broad-spectrum of diverse cutaneous PV strains. The obtained amplicons were directly sequenced. In the evaluated equine sarcoids, the four different sequences generated from obtained amplicons were shown to present 100% identity with the BPV13, the new BPV type identified from cutaneous warts of cattle. BPV13 was isolated from a cutaneous papilloma of the ear of a cow from a herd located in South region of Brazil. In order to amplify the total genome sequence of the BPV13, the PV genome was submitted to multiply-primed rolling circle amplification (RCA) technique. To confirm that the band was indeed PV DNA, PCR with degenerate primers specific for cutaneous PVs was performed on the RCA product. These PCR products were sequenced and based on the sequences generated, two primer sets for long template PCR were chosen in the partial obtained E1 and L1 sequences in order to amplify the most part of genome of the BPV13. The complete nucleotide sequences of the cloned long PCR products were determined by primerwalking sequencing, starting from the universal primers in the pCR4-TOPO vector. The remaining gaps in the sequences were determined by primer-walking directly on the RCA product. The complete genomic sequence of the BPV13 counts 7961 bp, with a GC content of 45.1%. BPV13 contains eight PV ORFs, coding for six early (E) proteins E1, E2, E4, E5, E6, and E7, and two late (L) proteins L1 and L2. The phylogenetic analysis using a concatenated nucleotide sequence alignment of E7, E1, E2, L2 and L1 ORFs of BPV13 isolated from a cow and other ungulate PVs showed that BPV13 is a member of the genus Deltapapillomavirus, closely related to BPV1and BPV2. The entire L1 ORF of BPV13 had 85.5-88.2% sequence identity, respectively, with Delta-PVs BPV1and 2. The identification of BPV13 in sarcoid lesions herein examined gives rise to the hypothesis that many PV isolates from sarcoid lesions previously diagnosed worldwide through PCR employing BPV1/2 specific primers were misclassified as BPV2, specially due to the direct sequencing of numerous amplicons were not performed. As it has been described for BPVs 1 and 2-induced lesions, it is tempting to suppose that BPV13 is also capable of inducing fibropapillomas in its natural hosts. The grouping of the BPV13 in Deltapapillomavirus genus, which lump fibropapilloma-inducing artiodactyls ruminant PVs together, as well as the finding of the E5 ORF in BPV13 genome and the lack of the pRB-binding motif in E7 encoded protein, support this hypothesis. PVs have a species-specific nature, with interspecies transmission being very rare event and only occurring between closely related species, such as infection of horses with BPV1 and 2. Like other Delta4 representatives, the novel BPV type was detected in association with cases of equine sarcoid in Brazilian herds. The characterization of novel BPVs improves our understanding on PV phylogeny and our interpretation of the described pathologies related to these viruses. Additional information about the molecular basis and epidemiology of characterized and uncharacterized BPVs will be needed to fully elucidate pathogenecity regarding diseases associated with BPV.