Search

OBJETIVO: O presente estudo realizou diagnóstico molecular e analisou fatores de risco associados à infecção de Leishmania spp. Hemoplasmas, Bartonella spp. e Pan-coronavírus em gatos do Nordeste do Brasil. MÉTODOS: Foram obtidas 127 amostras sanguíneas e 52 amostras de swabs retal de gatos domiciliados e errantes que foram extraídas e submetidas à PCR convencional (cPCR) com alvo na região kDNA de Leishmania spp. e do gene gltA de Bartonella spp.; e triadas por PCR em tempo real (qPCR) direcionado para o gene 16S rRNA de hemoplasmas. Amostras de RNA extraídas dos sobrenadantes de suspensões fecais foram submetidas à cPCR visando gene pan-coronavírus nsp12. RESULTADOS: A frequência de positividade foi de 12,6% (16/127) para hemoplasmas e 4,7% (6/127) para Leishmania spp., sendo identificada co-positividade para ambos os agentes em 0,79% (1/127) dos gatos avaliados. As amostras para Bartonella spp. e pan-coronavírus não foram amplificadas. Dos animais positivos para Leishmania e hemoplasma, 66,6% (4/6) e 81,2% (13/16), apresentaram maior frequência de sinais clínicos como: êmese, apatia e pirexia. CONCLUSÃO: Este estudo relata, pela primeira vez, coinfecção por Leishmania spp. e hemoplasmas em gatos da mesorregião do São Francisco, Nordeste do Brasil, expressando risco no subdiagnóstico de doenças felinas relevantes em saúde pública.

Of the 181 Brazilian bat species (Mammalia: Chiroptera), 46,7% occur in urban areas. However, little is known about the influence of these environments on species’ ecology, and few studies about urban bat community structure in Brazil have been published to date. The urbanization process, however, potentially lead to critical ecological changes in bat communities’ intra and interspecific pathogenic transmissions dynamics. Conversely, few published papers detected pathogens in urban bats. The present work aimed to perform bat captures in a preserved-rural-urban gradient from 2021 to 2022 to investigate how urbanization influences their richness, relative abundance, and activity degree. Most captured bats were Phyllostomidae, especially Sturnira lillium, Artibeus lituratus, A. fimbriatus, Glossophaga soricina, and Platyrrhinus lineatus, among others. From preserved-rural towards urban areas the lesser the bat richness, the higher the relative abundance of the captured bats. Noise level, luminosity and relative humidity correlated with bat abundance. The proportion of genders, sexually active bats and their size (weight, right forearm length, and body mass index) were stable throughout the investigation. Still, the proportion of pregnant females was higher in Spring and the number of juveniles in Summer, evidencing the seasonality of reproduction. No viruses (rabies and coronavirus), Salmonella spp. and Leptospira spp. were detected in the sample. Still, several Enterobacteria were isolated, evidencing a significant role of bats in the circulation of pathogens of medical and veterinary interest. These results are crucial in the pursuit of an harmonious coexistence between humans, bats and domestic animals in areas with different levels of anthropization.

AimsWe aimed to investigate the prevalence of rotavirus and coronavirus in dipterans that commonly inhabit the environment of dairy farms.Methods and ResultsWe collected 217 insect specimens from nine dairy farms, which were examined through hemi-nested RT-PCR followed by Sanger sequencing in search of VP1 and N genes for rotavirus and bovine coronavirus-BCoV, respectively. With a predominance of Muscidae (152/217 = 70%) 11 families of Diptera were identified. Rotavirus A (RVA) and betacoronavirus (BCoV) were detected in 14.7% (32/217) and 4.6% (10/217) of the dipterans, respectively. Sequencing of the amplicons was possible for 11.5% (25/217) of RVA and 0.5% (1/217) of BCoV, confirming the presence of these pathogens.ConclusionsOur findings highlight the role of dipterans as carriers of RVA and BCoV of great relevance for public and animal health.

Since 1998, with the occurrence of the last case of rabies in dogs or cats in São Paulo State related to the dog-specific rabies virus (RABV) lineage, 55 cases of rabies in these animals have been reported until 2021, and the vast majority (51) have been genetically characterized as belonging to the Desmodus rotundus/Artibeus lituratus lineage of RABV, However, it has not been demonstrated so far if there is the possibility to infer which of these bats acted as a source of infection in these cases through the genetic sequencing of isolated RABV. In order to characterize the Desmodus rotundus/Artibeus lituratus lineage aiming at the possibility of differentiating its isolates in RABV associated with each of these reservoirs, this research performed the DNA sequencing technique by amplification on the Illumina Miseq platform in 70 RABV isolates from the State of São Paulo, sent to the virology laboratory of the Pasteur Institute of São Paulo between 2006–2015, being 33 related to the hematophagous bat D. rotundus and 37 to the fruit bat A. lituratus. A genomic approach using phylogenetic and nucleotide identity analyses demonstrated that the isolates investigated in this study can be considered as belonging to the same genetic lineage of RABV and that in São Paulo State the D. rotundus/A. lituratus lineage is subdivided into at least four phylogenetic sub lineages, two associated with D. rotundus and two with A. lituratus, and that these results can be practically applied to the epidemiological surveillance of rabies in this state.

Lyssavirus rabies (RABV) is an RNA virus and, therefore, is subject to mutations due to low RNA polymerase replication fidelity, forming a population structure known as a viral quasispecies, which is the core of RNA viruses’ adaptive strategy. Under new microenvironmental conditions, the fittest populations are selected, and the study of this process on the molecular level can help determine molecular signatures related to virulence. Our aim was to survey gene signatures on nucleoprotein and glycoprotein genes that might be involved in virulence modulation during the in vitro evolution of RABV lineages after serial passages in a neuronal cell system with or without the presence of neutralizing antibodies based on replicative fitness, in vivo neurotropism and protein structure and dynamics. The experiments revealed that amino acids at positions 186 and 188 of the glycoprotein are virulence factors of Lyssavirus rabies, and site 186 specifically might allow the attachment to heparan as a secondary cell receptor, while polymorphism at position 333 might allow the selection of escape mutants under suboptimal neutralizing antibodies titers.

Bats are the second most diverse mammalian order worldwide. In the Neotropics, Brazil has one of the greatest bat richness, but species records are still very heterogeneously distributed, especially in Amazonia. Although this biome harbours 76% of Brazil’s bat fauna, basic knowledge on the distribution of Amazonian bats is far from well known, with intensive surveys at only a few sites and many areas without any records. The reduction of those gaps will improve the understanding of the bats’ distribution, with implications for management and conservation. Here we review the distribution of Trinycteris nicefori, Lionycteris spurrelli, Macrophyllum macrophyllum, and Dasypterus ega in the Brazilian Amazonia, presenting several new records for the region based both on literature review and on our own unpublished data. Our study contributes to the knowledge of the geographic distribution of these species, and the new records presented here help fill a gap in the distribution of bats in the western Brazilian Amazonia.

Rabies is a zoonotic neurological disease with 100% lethality. Some of the rare human patients who survived after multiple drug treatment have inherited severe sequelae. The objective of this study was to investigate the action of the transfection of antibodies against rabies in the central nervous system of mice as target therapy for rabies.Author summaryThe present study showed that after 48 h of RABV inoculation, mice injected by the intracerebral route with anti-RABV F(ab’)2 complexed with Bioporter® Protein Delivery Reagent (Genlantis) as a transfection agent, showing a morbidity/mortality rate of 30% with a minimum incubation period of seven days, while in the control group a significantly higher (p

Canine Distemper is a disease caused by Canine morbillivirus (CM), a pantropic virus that can affect the central nervous system (CNS), causing demyelination. However, the pathogenesis of this lesion remains to be clarified. Brain samples of 14 naturally infected dogs by CM were analyzed to evaluate the presence of oxidative stress and demyelination. RT-PCR assay was performed to confirm a diagnosis of canine distemper in the brain, immunohistochemistry anti-CM was used to localize the viral proteins in the tissue, and anti-4-hydroxy-2 nonenal (4-HNE) was a marker of a product of lipid peroxidation. The results showed the presence of viral proteins in the demyelinated area with the presence of 4-HNE. Our results suggest that the CM virus infection causes oxidative stress leading to lipid peroxidation, which causes tissue damage and demyelination. In conclusion, oxidative stress plays a significant role in canine distemper pathogenesis in the CNS.

Mast cell tumors (MCTs) are common neoplasms in dogs, and treatments for these diseases include surgery, polychemotherapy and targeted therapy with tyrosine kinase inhibitors. This study aimed to evaluate the response and the adverse events of treatment with imatinib mesylate (IM) compared to conventional therapy using vinblastine and prednisolone (VP) in canine cutaneous MCTs. Twenty-four dogs were included in the study; 13 animals were treated with IM and 11 with VP. Tumor tissue samples were submitted for histological diagnosis, grading and KIT immunostaining. The response to treatment was assessed by tomographic measurements according to VCOG criteria. Adverse events were classified according to VCOG-CTCAE criteria. The IM and VP groups had dogs with similar breeds, gender, ages, MCT localization, WHO stages and lymph node metastasis profiles. Most MCTs were grade 2/low and had KIT- patterns 2 and 3. The objective response rate (ORR) was significantly higher (30.79%) in the IM group then in VP group (9.09%). Adverse events (AE) in IM group were all grade 1, significantly different from VP. In conclusion, IM presented better ORR and less severe adverse events when compared to VP, representing a suitable option for the treatment of low-grade canine MCTs.

The screening of flu-like syndrome is difficult due to nonspecific symptoms or even oligosymptomatic presentation and became even more complex during the Covid-19 pandemic. However, an efficient screening tool plays an important role in the control of highly contagious diseases, allowing more efficient medical-epidemiological approaches and rational management of global health resources. Infrared thermography is a technique sensitive to small alterations in the skin temperature which may be related to early signs of inflammation and thus being relevant in the detection of infectious diseases. Thus, the objective of this study was to evaluate the potential of facial thermal profiles as a risk evaluator of symptoms and signs of SARs diseases, using COVID-19 as background disease. A total of 136 patients were inquired about the most common symptoms of COVID-19 infection and were submitted to an infrared image scanning, where the temperatures of 10 parameters from different regions of the face were captured. We used RT-qPCR as the ground truth to compare with the thermal parameters, in order to evaluate the performance of infrared imaging in COVID-19 screening. Only 16% of infected patients had fever at the hospital admission, and most infrared thermal variables presented values of temperature significantly higher in infected patients. The maximum eye temperature (MaxE) showed the highest predictive value at a cut-off of >35.9°C (sn = 71.87%, sp = 86.11%, LR+ = 5.18, LR- = 0.33, AUC = 0.850, p < 0.001). Our predictive model reached an accuracy of 86% for disease detection, indicating that facial infrared thermal scanning, based on the combination of different facial regions and the thermal profile of the face, has potential to act as a more accurate diagnostic support method for early COVID-19 screening, when compared to classical infrared methods, based on a single spot with the maximum skin temperature of the face.

The early use of antimicrobial therapy has been introduced in many farms to prevent diarrhea and respiratory disease in young calves; however, there is controversy about whether this practice has a beneficial effect on the health of these animals. This study evaluated the influence of the early use of antimicrobials on the health and performance of neonatal Holstein calves. Twenty-six Holstein calves were screened and divided into two groups, according to the administration (ATB+), or not (ATB-) of tulathromycin (2.5mg/kg, subcutaneously) within the first 12 hours of life. Calves were evaluated by general clinical examination, fecal score, respiratory score, and external palpation of the umbilical region, besides fecal output of dry matter. Anemia was determined by using an automatic system and, also, using a commercial kit for iron dosage. Diarrhea was diagnosed by a centrifuge-flotation technique using a sugar solution (Cryptosporidium) and multiplex semi-nested RT-PCR (rotavirus/coronavirus). The performance of the calves was estimated by Daily Weight Gain (DWG). The young dairy calves were evaluated within 12 hours of birth (≤12h) and at 3-5th (D3-5), 7-9th (D7-9), 13-15th (D13-15), 20-23rd (D20-23), and 27-30th (D27-30) days of life. No difference was noted between the ATB+ and ATB- groups concerning heart rate, respiratory frequency, and rectal temperature. Erythrogram showed a higher frequency of anemia in ATB- group (P=0.016) at the D3-5 check-up; lower values of serum iron were also observed simultaneously (P=0.051). Thirteen cases of respiratory disease were detected during this study; however, no significant difference was observed between the groups in this regard. The frequency of diarrhea (fecal score 2-3) was high in both groups, peaking at D13-D15. No differences were noted between the groups regarding the frequency of diarrhea when considering the dry fecal matter. The predominant etiological agent for diarrhea was Cryptosporidium spp.. The DWG was similar between groups, with maximum weight reduction on D13-15. The administration of tulathromycin in prophylactic dose (2.5mg/kg) at birth decreased the frequency of anemia but did not influence weight gain or the prevalence of diarrhea.

We obtained the complete sequence of a novel poxvirus, tentatively named Brazilian porcupinepox virus, from a wild porcupine (Coendou prehensilis) in Brazil that had skin and internal lesions characteristic of poxvirus infection. The impact of this lethal poxvirus on the survival of this species and its potential zoonotic importance remain to be investigated.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), responsible for the coronavirus disease 2019 (COVID-19), is the causative infectious agent of the current pandemic. As researchers and health professionals are still learning the capabilities of this virus, public health concerns arise regarding the zoonotic potential of SARS-CoV-2. With millions of people detected with SARS-CoV-2 worldwide, reports of companion animals possibly infected with the virus started to emerge. Therefore, our aim is to review reported cases of animals naturally infected with SARS-CoV-2, particularly companion pets, shedding light on the role of these animals in the epidemiology of COVID-19.

Feline immunodeficiency virus (FIV) is an important pathogen of domestic and wild felids. Although serological tests suggest the presence of FIV in cats from Colombia, no molecular characterization has been reported. Here, we describe the near-complete genome of FIV subtype A from a Colombian domestic cat.

El lector está recibiendo un Manual Técnico de Operaciones desarrollado por investigadores de la Universidad de São Paulo (USP, Brasil) y de la Universidad de Cambridge (Reino Unido). Este documento pretende ofrecerle a los técnicos extensionistas, un conjunto de orientaciones que pretenden disminuir el riesgo de propagación del Coronavirus (Sars.CoV2), para las diversas situaciones que puedan presentarse en su trabajo junto a los productores rurales, funcionarios y familiares; que suceden en las rutinas productivas de las granjas de cerdos visitadas. Debido a la pandemia global de la COVID-19, es importante que medidas de bioseguridad sean tomadas para evitar la contaminación de los técnicos, productores de cerdos, su familia y funcionarios, en busca de asegurar y mantener la salud de esas personas en sus territorios rurales. Complementariamente, este manual técnico también pretende contribuir con el mantenimiento de los procesos productivos de los mataderos. Actualmente la ciencia está todos los días haciendo nuevos descubrimientos sobre la COVID-19, y, por lo tanto, el contenido de este manual será actualizado en la medida de lo posible.

Este Manual Técnico Operacional foi desenvolvido por pesquisadores da Universidade de São Paulo e da Universidade de Cambridge (UK), visando ofertar, ao leitor técnico extensionista e também aos interessados no tema, um conjunto de orientações que busca diminuir o risco de propagação do Coronavírus (Sars-CoV-2), nos mais variados tipos de encontros de trabalho que acontecem nas rotinas produtivas das granjas de suínos junto dos produtores rurais, funcionários e familiares. Devido à pandemia global da COVID-19, é importante que medidas de biossegurança sejam tomadas para evitar a contaminação dos técnicos, produtores de suínos, seus familiares e funcionários, buscando assegurar a manutenção da saúde dessas pessoas nos seus territórios rurais. De forma complementar, este manual técnico também visa contribuir com a manutenção dos processos produtivos dos frigoríficos. De acordo com as normas vigentes de biossegurança, este documento oferta um conjunto de informações sobre procedimentos interacionais, os quais deverão ser internalizados e adotados nas rotinas produtivas de cada granja de suínos. A ciência revela, todos os dias, novas descobertas e novos conhecimentos e, portanto, o conteúdo deste manual será atualizado na medida do possível.

Cetacean morbillivirus (CeMV, family Paramyxoviridae) is a re-emergent pathogen associated with severe epizootic outbreaks causing high mortality among cetaceans worldwide. Recently, CeMV caused an unusual mortality event of Guiana dolphins (Sotalia guianensis) in Brazil. Partial sequence of the viral phosphoprotein (P) gene showed that the Guiana dolphin morbillivirus (GDMV) might represent a new lineage of CeMV. This study aimed to develop a molecular technique to detect the most common CeMV strains known to circulate in the Atlantic Ocean: GDMV, Dolphin morbillivirus (DMV) and Pilot-whale morbillivirus (PWMV). A sensible real-time reverse transcription polymerase chain reaction (RT-qPCR) method based on intercalating dye, targeting the P gene was described. This assay successfully detected GDMV, PWMV and DMV from field samples. Its performance was compared to a RT-qPCR method that specifically detects GDMV. Both assays had high sensibility and excellent intra- and inter-assay reproducibility. A total of 109 field samples from 32 Guiana dolphins were screened for CeMV by conventional RT-PCR in parallel with the RT-qPCR assay. The detection rate increased from 32% to 60% by use of the novel RT-qPCR. The RT-qPCR assay described herein allows rapid and sensitive detection of Atlantic CeMV strains, and is potentially suitable for screening of CeMV globally.

Introduction: Staphylococci are the most important agents associated with bovine mastitis. This study aimed at characterizing resistance factors to antimicrobials in Staphylococcus spp. isolated from the milk of cows with subclinical mastitis. Methodology: In vitro resistance of 243 Staphylococcus spp. isolates to antimicrobials commonly used in clinical practice was evaluated. The detection and expression of genes encoding resistance mecA (gene encoding penicillin binding protein 2a) mecALGA251 (mecA homologue), blaZ (gene encoding penicillin resistance), femA and femB (genes encoding essential factors - A and B - for the expression of methicillin resistance) and aacA-aphD (gene encoding for a bifunctional enzyme that confers resistance to gentamicin) using PCR and RT-PCR was investigated. Results: One or more genes encoding resistance to different antimicrobials were detected in 184 Staphylococcus spp. samples. The femA and femB genes were the most frequent. Regarding the variables’ detection (N = number of strains) and expression (% of strains), the following results were obtained: blaZ (N = 40 – 82.5%), femA (N = 147 – 47.6%), aacAaphD (N = 30 – 43.3%), femB (N = 138 – 29.7%), mecA (N = 33 – 27.3%), mecALGA251 (N = 01 – 0.0%). There was a higher occurrence of phenotypic resistant strains for amoxicillin, ampicillin and penicillin in isolates positive for detection and/or expression of blaZ gene when compared with the other genes. Conclusions: The present study provides new information on genotypic traits of Staphylococcus isolates from bovine subclinical mastitis especially regarding the evaluation of expression of genes associated with antimicrobial resistance in Staphylococcus spp. using molecular tools.

A modified TaqMan real-time polymerase chain reaction targeting a 138 bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleotide sequencing analysis. The overall agreement between the assays was 94.8% (confidence interval [CI] 95% 88–100%). The newly developed assay presented 91.6% (CI 95% 71.5–98.5%) relative sensitivity (22[+] lipl32 PCR/24[+] 16S RNA and sequencing), 100% (CI 95% 96.3–100%) relative specificity and 98.7% accuracy (CI 95% 94.8–100%). The lipl32 assay was able to detect and quantify at least 10 genome equivalents/reaction. DNA extracted from 17 pathogenic Leptospira spp., 8 intermediate/saprophytic strains and 21 different pathogenic microorganisms were also tested using the lipl32 assay, resulting in amplification exclusively for pathogenic leptospiral strains. The results also demonstrated high intra and inter-assay reproducibility (coefficient of variation 1.50 and 1.12, respectively), thereby qualifying the newly developed assay as a highly sensitive, specific and reliable diagnostic tool for leptospiral infection in dogs using urine specimens.

The equine influenza virus (EIV) H3N8 subtype is responsible for all EIV outbreaks worldwide while the H7N7 subtype is less pathogenic and is considered extinct as it has not been confirmed in outbreaks since 1980. Although EIV is enzootic in Brazil, few reports describe the actual EIV antibody status in the country. The aims of this study were: - to evaluate the efficiency of different serum treatments described by the World Organisation for Animal Health (OIE) and the World Health Organization (WHO) to remove non-specific haemagglutination inhibitors for the haemagglutination inhibition (HI) assay for EIV - to evaluate the presence of EIV antibodies by HI, enzyme-linked immunosorbent assay and agar gel immunodiffusion in 83 non-vaccinated equines from Sao Paulo State - to evaluate a strategy to better analyse equine sera for EIV antibodies. Although there was no statistical difference among treatments, receptor-destroying enzyme treatment followed by chicken erythrocyte adsorption showed more consistent results, which corroborate the OIE and WHO recommendation to use this treatment preferentially. The HI results suggest equine H3N8 virus circulation among the animals tested from Sao Paulo State. The algorithm suggested here could be used to guide antibody detection against equine influenza virus in equines, improving the test specificity by aiming to avoid false positive results.

Made available in DSpace on 2018-12-11T17:14:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-09-01 We describe molecular testing for felid alphaherpesvirus 1 (FHV-1), carnivore protoparvovirus 1 (CPPV-1), feline calicivirus (FCV), alphacoronavirus 1 (feline coronavirus [FCoV]), feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and canine distemper virus (CDV) in whole blood samples of 109 free-ranging and 68 captive neotropical felids from Brazil. Samples from 2 jaguars (Panthera onca) and 1 oncilla (Leopardus tigrinus) were positive for FHV-1; 2 jaguars, 1 puma (Puma concolor), and 1 jaguarundi (Herpairulus yagouaroundi) tested positive for CPPV-1; and 1 puma was positive for FIV. Based on comparison of 103 nucleotides of the UL24-UL25 gene, the FHV-1 sequences were 99–100% similar to the FHV-1 strain of domestic cats. Nucleotide sequences of CPPV-1 were closely related to sequences detected in other wild carnivores, comparing 294 nucleotides of the VP1 gene. The FIV nucleotide sequence detected in the free-ranging puma, based on comparison of 444 nucleotides of the pol gene, grouped with other lentiviruses described in pumas, and had 82.4% identity with a free-ranging puma from Yellowstone Park and 79.5% with a captive puma from Brazil. Our data document the circulation of FHV-1, CPPV-1, and FIV in neotropical felids in Brazil. Departments of Preventive Veterinary Medicine and Animal Health School of Veterinary Medicine and Animal Science University of São Paulo Departments of Pathology School of Veterinary Medicine and Animal Science University of São Paulo Jaguar Conservation Fund Instituto Onça-Pintada Institute for the Conservation of Neotropical Carnivores Instituto Pró-Carnívoros Institute for Ecological Research Instituto de Pesquisas Ecológicas (IPE) Department of Microbiology and Immunology Biosciences Institute State University of São Paulo (UNESP) Júlio de Mesquita Filho Brazilian Institute for Conservation Medicine Instituto Brasileiro para Medicina da Conservação (TRÍADE) Chico Mendes Institute for Biodiversity Conservation ICMBio Department of Microbiology and Immunology Biosciences Institute State University of São Paulo (UNESP) Júlio de Mesquita Filho

Rotaviruses are members of the family Reoviridae and are a common cause of acute diarrhea in many mammalian and avian species. They are non-enveloped icosahedral particles and their genome comprises 11 segments of double-stranded RNA, which encodes six structural proteins (VP1–4, VP6–7) and six nonstructural proteins (NSP1–6). Genotypes are defined based upon the diversity found in these genes and viral characterization plays a central role on epidemiological studies and prevention. Here we investigate the distribution of Brazilian RVAs genotypes in 8 chicken samples collected between 2008 and 2015 from different regions by RT-PCR, partial (Sanger) nucleotide sequencing and phylogenetic analysis from all rotavirus genes. Although the identified genotypes were typical from avian host species, when analyzed together, they form novel genetic constellations: G19-P[31]-I11-R6-C6-M7-A16-N6-T8-E10-H8 and G19-P[31]-I4-R4-C4-M4-A16-N4-T4-E4-H4. This study highlights that avian rotaviruses are widespread among commercial farms in Brazil, and the co-circulation of at least two different genomic constellations indicates that may present a way bigger genetic variability, that can be increased by the possible transmission events from other birds, lack of specific preventive measures, as well as the different viral evolution mechanisms. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s42770-020-00259-4) contains supplementary material, which is available to authorized users.

The vampire bat (Desmodus rotundus) plays a crucial role in the maintenance and transmission of the rabies virus to humans and livestock, impacting public health and economic production. Its importance lies not only in its capacity to transmit the virus but also in its ability to adapt to anthropic changes, as expressed in its wide geographic distribution in Latin America. Deforestation, livestock intensification, and other human activities have reduced the abundance of its wild prey but have also provided new and abundant shelter and foraging resources for the vampire bats. We used radiotelemetry to evaluate relations between topography, feeding site choice, and foraging behavior in southeastern Brazil, where three occupied D. rotundus roosts, out of 11 possible, were systematically monitored throughout a 1-year period once every 2 months. Sixty-two vampire bats were captured; biometric data were collected and 44 radiotransmitters were installed in adult individuals, producing telemetry data that were recorded in VHF receptors installed in the farms. Elevation of the roosts was related to the farms attacked by the vampire bats. Understanding the use of the environment and resources by vampire bats is critical to improving rabies control aiming at the reduction of disease impacts. From the perspective of the official veterinary service, telemetry would be ineffective as a rabies surveillance system due to the costs and limitations of the available technology. However, livestock rabies control measures would be greatly improved if ecological characteristics of the vampire bat were considered.

Avian coronavirus (AvCoV) infects a range of tissues in chickens and several other avian species. Although the viruscan be isolated in chicken embryos, only a few strains of the 6 genotypes/33 lineages can grow in cell lines, with theBeaudette strain (GI-1 lineage) being the most used for in vitro studies. Considering the differences between celllines and chicken embryos as habitats for AvCoV, this study aimed to assess the diversity of the genes coding forthe nonstructural protein 3 (nsp3) and spike envelope protein (S) after serial passages in BHK-21 and Vero cells.After 14 passages of an embryo-adapted Beaudette strain, the virus loads fluctuated in both cell lines, with the highestloads being 8.72 log genome copies/µL for Vero and 6.36 log genome copies/µL for BHK-21 cells. No polymorphismswere found for nsp3; regarding S, not only aa substitutions (Vero: 8th passage A150S, and 14th S150A; BHK-21: 4thS53F, 8th F53Y, and 8th S95R), but also minor variants could be detected on chromatograms with fluctuating intensities.As the regions of these aa substitutions are within the receptor-binding domain of S, it can be speculated that differencesin cell receptors between Vero and BHK-21 cells and the speed of cell death led to the selection of different dominantstrains, while the stability of nsp3 supports its function as a protease involved in AvCoV replication. In conclusion,AvCoV quasispecies evolution is influenced by the biological model under consideration, and a gradual transition isseen for minor and major variants. O Coronavírus aviário AvCoV infecta uma variedade de tecidos de galinhas e de outras espécies aviárias. Apesarde este vírus poder ser isolado em ovos embrionados de galinha, apenas alguns dos 6 genótipos / 33 linhagenspodem crescer em cultivo celular, sendo a cepa Beuadette (linhagem GI-11) a mais utilizada para estudos in vitro.Considerando as diferentes linhagens celulares e ovos embrionados como habitats para o AvCoV, este estudo tevepor objetivo estudar a diversidade de genes que codificam para a proteína não-estrutural 3 (nsp3) e espícula (S)após passagens seriadas em células BHK-21 e VERO. Após 14 passagens, de uma amostra Beuadette adaptada aovos embrionados, os títulos virais variaram em ambas as células, com os maiores títulos sendo de 8,72 log cópiasgenômicas/µL para Vero e 6,36 cópias genômicas/µL para BHK-21. Nenhum polimorfismo foi encontrando paransp3. Considerando a proteína S, não somente foram encontradas substituições de aminoácidos (Vero: 8apassagemA150S e 14apassagem S150A; BHK-21: 4apassagem S53F, 8apassagem F53Y e S95R), mas também, variantessubconsensuais foram detectadas pelos cromatogramas com intensidades flutuantes. Uma vez que as regiões destesaa se encontram no domínio de ligação de receptor de S, pode-se especular que diferenças em receptores celularesentre Vero e BHK-21, além da velocidade da morte celular, levaram à seleção de diferentes cepas dominantes,enquanto que a estabilidade de nsp3 concorda com sua função como protease com papel na replicação de AvCoV.Como conclusão, a evolução de quase-espécies de AvCoV é influenciada pelo modelo biológico sob consideraçãoe uma transição gradual é vista para variantes dominantes e subdominantes.&nbsp

Rabies is a fatal zoonotic infection of the central nervous system of mammals and has been known to humans for millennia. The etiological agent, is a neurotropic RNA virus in the order Mononegavirales, family Rhabdoviridae, genus Lyssavirus. There are currently accepted to be two cycles for rabies transmission: the urban cycle and the sylvatic cycle. The fact that both cycles originated from a common RABV or lyssavirus ancestor and the adaptive divergence that occurred since then as this ancestor virus adapted to a wide range of fitness landscapes represented by reservoir species in the orders Carnivora and Chiroptera led to the emergence of the diverse RABV lineages currently found in the sylvatic and urban cycles. Here we study full genome phylogenies and the time to the most recent common ancestor (TMRCA) of the RABVs in the sylvatic and urban cycles. Results show that there were differences between the nucleotide substitution rates per site per year for the same RABV genes maintained independently in the urban and sylvatic cycles. The results identify the most suitable gene for phylogenetic analysis, heterotachy among RABV genes and the TMRCA for the two cycles.

Alphacoronavirus 1 (subgenus Tegacovirus, genus Alphacoronavirus, family Coronaviridae), which encompasses transmissible gastroenteritis virus (TGEV), feline coronavirus (FCoV) and canine coronavirus (CCoV), is an important pathogen that can cause severe gastroenteritis and is distributed worldwide. CCoV has two different genotypes: CCoV type I, which has a high identity with FCoV-I, and CCoV type II, which is divided into two subtypes, CCoV IIa (pantropic) and CCoV IIb, which is related to FCoV-II and has been involved in multiple recombination events. Between 2014 and 2018, 43 fecal samples from puppies and young dogs under 1 year of age with hemorrhagic enteritis and from 5 cats under 2 years of age with ascites or thoracic effusion were collected by a private veterinary practice in Bogotá, Colombia. A screening for Coronavirus via RT-PCR (nsp12) and PCR amplification of Canine protoparvovirus (VP1) revealed 27.1% (13/49) and 72.9% (35/49) positive samples, respectively. Positive samples for coronavirus were tested for M, N, S and the sequences grouped in the FCoV, CCoV-I and CCoV-IIb clusters that were distant from the pantropic type (IIa). The N gene formed two clusters, one exclusively with samples from this study in subtype II and another with strains in subtype I. For gene S (subtype I), the samples clustered with the Brazilian samples, while samples positive for S subtype IIb grouped into a cluster distinct from the other reference sequences. The prevalence of coronaviruses identified in this study is within the range reported by different countries worldwide., Genetics; Microbiology; Virology; Biocomputational method; Viral disease; Viral genetics; Phylogeny; Epidemiology; Coronavirus; Alphacoronavirus 1; Dogs; Cats; Colombia; Phylogenetic relationship.

In this study, we describe the viral composition of adult Antricola delacruzi ticks collected in a hot bat cave in the state of Rondônia, Western Amazonia, Brazil. A. delacruzi ticks, are special, compared to many other ticks, in that they feed on both bats (larval blood feeding) and bat guano (nymphal and adult feeding) instead of feeding exclusively on vertebrate hosts (blood feeding). Considering this unique life-cycle it is potentially possible that these ticks can pick up/be infected by viruses not only present in the blood of viremic bats but also by virus shed through the bat guano. The viral metagenomic investigation of adult ticks showed that single-stranded negative-sense RNA viruses were the dominant group of viruses identified in the investigated ticks. Out of these, members of the Nairoviridae family were in clear majority constituting 88% of all viral reads in the data set. Genetic and phylogenetic analyses indicate the presence of several different orthonairoviruses in the investigated ticks with only distant relationship to previously described ones. In addition, identification of viral sequences belonging to Orthomyxoviridae, Iflaviridae, Dicistroviridae, Polycipiviridae, Reoviridae and different unclassified RNA viruses showed the presence of viruses with low sequence similarity to previously described viruses.

This paper expands on a previous report about coronaviruses in quail. After surveillance carried out in 2009 and 2010, some farmers started vaccinating quail with the Massachusetts avian infectious bronchitis virus serotype. The samples for this study were collected in 2013 from São Paulo state in southeastern Brazil. Pools of trachea, lungs, reproductive tract, kidneys, and enteric contents from quail and laying hens kept in the same farms and from quail-only farms as well as from both healthy birds and those showing infectious bronchitis-like symptoms were sampled in this study. The samples were screened using nested RT-PCR targeting the 3'-untranslated region of the Gammacoronavirus genus. Based on the DNA sequence for the RNA-dependent RNA polymerase (RdRp) gene, the strains isolated from quail clustered within either the Gammacoronavirus or Deltacoronavirus genus, and sequences from both genera were found in one quail sample. The phylogeny based on the partial S1 subunit sequence showed that the gammacoronaviruses detected in quail and layers belonged to the Brazil type. These results suggest that quail are susceptible to Gammacoronavirus and Deltacoronavirus viruses and indicate that the Massachusetts vaccination was not controlling IBV in quail or chickens.

Caprine arthritis encephalitis (CAE) is a multisystem infectious disease caused by a small ruminant lentivirus that is widely disseminated in the goat flock of southeastern Brazil. Little information about its variants is available in Brazil. Phylogenetic analysis was performed on the sequences of 14 clinical isolates of CAEV that obtained from mammary glands, lung, brain, milk and blood of goats with any clinical form of the disease in Sao Paulo State, Brazil. Part of the pol gene of CAEV was amplified by nested-PCR and sequenced. These sequences were compared with 42 reference strains previously described and commonly used in phylogenies. The sequences derived from this study were more related to caprine strains than to ovine strains. Additionally, subtype B of the CAEV group was detected in different clinical samples with distinct clinical symptoms. Subgroup B1 was the predominant genotype. A subgroup C-like was isolated from animals that presented interstitial indurative mastitis. Animals from which the sequences belonged to this subgroup presented interstitial mastitis induration, chronic arthritis and chronic interstitial pneumonia. Subgroup C sequences have not been reported in Brazil previously, while circulating subgroup B1 variants were already known to be present in the country. This study provides new information about CAEV variants in Brazil, since we isolated and reported the subgroup C-like. A variant from B1 subgroup was the predominant genotype. More studies are necessary in order to confirm the presence of CAEV genotype C or even a new genotype. CAEV is an important disease of goat production and all knowledge is necessary to understand its epidemiology and, prevent this illness.

With the raise of SARS-CoV-2 in late 2019 and the associated COVID-19 pandemics, much of the knowledge gathered from decades on research on other coronavirus species that use humans and other animals as hosts is invaluable to help counteract the burden of this new disease. This review ais to bring the highlights on coronavirus Biology and the most frequent diseases they might cause, serving rather as an introduction to the field.

Bovine coronavirus (BCoV) is one of the main aetiological agents of gastroenteritis in calves, causing significant economic damage to livestock. This study aims to characterise BCoV genetically on the basis of the N gene. A total of 114 faecal samples from beef and dairy calves with or without clinical symptoms of diarrhoea from five Brazilian states (São Paulo, Minas Gerais, Santa Catarina, Mato Grosso and Bahia) were evaluated between 2008 and 2015 by technique of Semi‐nested RT‐PCR for gene N and genealogical analysis. Of the 114 samples analysed, 14.91% (17/114) were positive. BCoV was detected in 22.72% (10/44) of the animals with diarrhoea and in 10% (7/70) of asymptomatic animals. BCoV was identified in calves from rural properties located in all of the regions sampled. Genealogical analysis showed that the Brazilian sequences of BCoV for the gene which codes for the N protein can be broken down into two distinct clusters, and the samples from this study were closely linked to Asian strains. These results contribute to the molecular characterization of BCoV in Brazil and are the first report of the circulation of BCoV in the states of Santa Catarina and Bahia.

Introduction: The identification of species that act as reservoirs or hosts of zoonotic agents is essential for control and epidemiological surveillance of the important illness in public health. Identification of the reservoirs for zoonoses can help to clarify how the pathogens are maintained in nature, leading to more effective disease control and avoiding indiscriminate extermination of wild animals.

Made available in DSpace on 2018-12-11T17:31:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Diarrhea is a major clinical problem affecting foals up to 3 months of age. The aim of this study was to identify enteric microorganisms involved in monoinfections and coinfections and the associated virulence factors in healthy and diarrheic foals. Diarrheic (D) (n=56) and nondiarrheic (ND) foals (n=60) up to three months of age were studied. Fecal samples were analyzed for identification of infectious agents (microbiological culturing, molecular techniques, and microscopic analyses). Escherichia coli fimH (30% versus 25%), Salmonella spp. (25% versus 7%), Strongyloides westeri (25% versus 25%), Clostridium perfringens type A (21% versus 10%), E. coli ag43 (20% versus 35%), Strongylus (11% versus 18%), and vapA-positive Rhodococcus equi (5% versus 2%) were the most frequent enteric pathogens detected in D and ND foals, respectively. The frequency of toxin A-positive C. perfringens was significantly increased in the D (p=0.033) compared with the ND animals. R. equi strains harboring virulent plasmids were also identified (VapA 85-kb type I and VapA 87-kb type I) in D and ND foals. Coinfections were observed in 46% of the D and 33% of the ND foals. Our results demonstrate the great diversity of enteric pathogens, virulence factors, and coinfections involved in enteric infections of foals. Department of Veterinary Clinical Sciences Universidade Estadual Paulista (UNESP) Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP) Department of Preventive Veterinary Medicine Federal University of Minas Gerais (UFMG) Department of Genetics Evolution and Bioagents Campinas State University (Unicamp) Department of Preventive Veterinary Medicine University of S�o Paulo Department of Animal Hygiene Kitasato University Department of Veterinary Clinical Sciences Universidade Estadual Paulista (UNESP) Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP)

Many native bird species are currently considered rare in Brazil because they have been indiscriminately collected by animal traffickers and commercialized, leading to dwindling numbers in their natural habitats. Confiscated animals are at times destined for reintroduction programs that must ensure these animals do not pose a risk to native populations. Healthy or sick wild passerines may carry a great diversity of microorganisms. Therefore, knowledge of the sanitary status of confiscated animals destined for reintroduction is critical to assess whether these animals act as microorganism carriers and to investigate the epidemiology of transmissible diseases, a crucial aspect for animal and human health preservation. This study examined the occurrence of aerobic and facultative anaerobic bacteria and fungi in cloacal swabs collected from wild confiscated passerines intended for reintroduction programs. In vitro susceptibility tests of the most frequent isolates as well as studies of the molecular aspects of Escherichia coli isolates were also performed. There was microorganism growth in 62.5% of 253 samples. The microorganisms that were most frequently isolated were Staphylococcus spp. (15.0%), Micrococcus spp. (11.5%), E. coli (10.7%) and Klebsiella spp. (10.7%). Fifteen bacteria genera and seven fungi genera were isolated. Multidrug-resistance to antimicrobials was observed in Staphylococcus spp., Micrococcus spp., E. coli and Klebsiella spp. isolates. The high occurrence of Enterobacteria observed is possibly related to the sanitary conditions in which confiscated animals are usually kept. One E. coli sample (out of 27 isolates) was positive for the S-fimbrial adhesion encoding gene (sfa). Considering the low occurrence of genes that encode virulence factors, confiscated passerines may represent a low risk for the potential transmission of EPEC, APEC, UPEC and NMEC isolates to other animals or humans. The potential risk of intra- or inter-specific transmission of multidrug-resistant isolates and the introduction of these microorganisms into the environment must be considered, although there are still therapeutic alternatives for treatment of these animals among the antimicrobials which were tested. The stress and poor hygiene conditions imposed on animals during trafficking may have caused their contamination by multidrug-resistant agents transmitted by humans or by the precarious environment to which they were subjected. Risks related to the dissemination of Salmonella spp., Cryptococcus spp. and Candida spp. are low when reintroduction programs are considered.

A raiva é uma zoonose que afeta o sistema nervoso central, de evolução aguda e fatal, mantida em mamíferos e conhecida há milênios. Presente na América, Europa, África e Ásia, tem como agente etiológico o vírus da raiva (RABV), um vírus RNA neurotrópico, pertencente à ordem Mononegavirales, família Rhabdoviridae, gênero Lyssavirus., o qual é composto por quatorze espécies. Entre os Lyssavirus, o RABV é o mais amplamente distribuído mundialmente e tem maior importância epidemiológica dada sua associação com um maior número de casos de encefalite por Lyssavirus em humanos em relação às outras espécies. São admitidos dois ciclos de transmissão para a raiva, o ciclo urbano e o ciclo silvestre. O ciclo urbano ou terrestre tem o cão como principal reservatório e transmissor do vírus para outros cães, outros animais domésticos e para o homem, enquanto o ciclo silvestre ou aéreo é mantido por diferentes mamíferos silvestres e quirópteros. A origem comum dos dois ciclos do RABV á partir de um RABV ou Lyssavirus ancestral e a divergência adaptativa ocorrida desde então, causada pela adaptação de tal vírus em paisagens adaptativas tão variadas e distintas representadas pelas ordens Carnivora e Chiroptera, levaram ao surgimento das diversas linhagens encontradas nos ciclos terrestre e aéreo. Sendo assim, com o objetivo de se estudar as diferenças geradas nos RABV dos ciclos aéreo e terrestres devido a sua evolução em paralelo nestas duas ordens foram analisadas 159 sequências genômicas do RABV (59 do ciclo terrestre e 100 do ciclo aéreo), sendo que 21 destas sequências foram obtidas neste estudo e representam oito linhagens de RABV existentes no Brasil e cinco destas linhagens de RABV tiveram seus genomas sequenciados pela primeira vez. Foram analisados aspectos como as diferentes taxas de substituição de nucleotídeos por sítios (heterotaquia) entre os mesmos genes do RABV mantidos no ciclo aéreo e terrestre, análise do melhor gene para a realização de estudo filogenéticos confiáveis para o RABV, tempo de divergência entre os ciclos, padrões de variabilidade genética e vieses quanto ao uso preferencial de códons em cada ciclo. Como resultado, concluí-se que a divergência adaptativa ocorrida entre os dois ciclos do RABV fez com que alguns aspectos evolutivos de seu genoma apresentem padrões diferentes de acordo com o ciclo do RABV analisado. Rabies is a zoonosis that affects the central nervous system, showing an acute and fatal evolution, occurring in mammals and known for millennia. Present in America, Europe, Africa and Asia, its etiological agent is Rabies virus (RABV), a neurotropic RNA virus in the order Mononegavirales, family Rhabdoviridae, genus Lyssavirus, composed by fourteen species. Amongst the lyssaviruses, RABV is the most widely spread worldwide and has a higher epidemiological importance due to its association to a higher number of cases of encephalitis. Two cycles are accepted for rabies transmission, the urban and the wild ones. In the urban (or terrestrial) cycle, dogs are the main reservoirs and transmitters of the virus to other dogs, other domestic animals and to humans, while in the wild (or aerial) cycle bats are the reservoirs. The common origin of both cycles from an ancestor RABV or lyssavirus and the adaptive divergence that occurred since then, caused by the adaptation of this ancestor virus to a wide range of adaptive landscapes represented by the orders Carnivora and Chiroptera led to the emergence of diverse RABV lineages currently found in the aerial and terrestrial cycles. Thus, aiming to study differences found in RABV lineages from the aerial and terrestrial cycles due to their parallel evolution in these two orders, 159 genomic sequences of RABV (59 from the terrestrial and 100 from the aerial cycles) were analyzed, being 21 of these sequences referent to eight lineages of RABV found in Brazil sequenced in this study and five of these eight lineages of RABV had their genomes sequenced for the first time The study included the per site nucleotide substitution rate differences (heterotachy) between the same genes RABV maintained in the aerial and terrestrial, survey of the most suitable gene for phylogenetic analysis, time of divergence between the two cycles, patterns of genetic variability and codon usage bias. As a conclusion, the adaptive divergence occurred between the two cycles caused some evolutionary aspects of RABV genome to show an intricate cycle-specific evolutionary pattern.

Avian coronavirus, the causative agent of avian infectious bronchitis, occurs as multiple genotypes and lineages, and full genomes are not available for the majority of them. This paper reports the (previously unknown) complete genome sequence of strain D274 of this virus (27,599 nucleotides), isolated from chickens in The Netherlands in 1979.

Made available in DSpace on 2019-10-04T12:36:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-09-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Feline immunodeficiency virus (FIV) has worldwide distribution; nevertheless, only a few FIV genomes from domestic cats are available. This is the first report of a nearly complete genome of FIV from a Brazilian cat (8,967 nucleotides [nt]), including the entire coding region and the 3' untranslated region. Univ Sao Paulo, Dept Prevent Vet Med & Anim Hlth, Sch Vet Med, Sao Paulo, SP, Brazil Sao Paulo State Univ UNESP, Dept Microbiol & Immunol, Inst Biosci, Botucatu, SP, Brazil Sao Paulo State Univ UNESP, Inst Biotechnol IBTEC, Botucatu, SP, Brazil Sao Paulo State Univ UNESP, Dept Microbiol & Immunol, Inst Biosci, Botucatu, SP, Brazil Sao Paulo State Univ UNESP, Inst Biotechnol IBTEC, Botucatu, SP, Brazil CNPq: 301225/2013-3 CAPES: 1841/2016 FAPESP: 2014/13532-3

This paper reports on an investigation of the role of codon usage evolution on the suggested bovine-to-human spillover of Bovine coronavirus (BCoV), an enteric/respiratory virus of cattle, resulting in the emergence of the exclusively respiratory Human coronavirus OC43 (HCoV-OC43). Analyses based on full genomes of BCoV and HCoV-OC43 and on both human and bovine mRNAs sequences of cholecystokinin (CCK) and surfactant protein 1 A (SFTP1-A), representing the enteric and respiratory tract codon usage, respectively, have shown natural selection leading to optimization or deoptimization of viral codon usage to the human enteric and respiratory tracts depending on the virus genes under consideration. A higher correlation was found for the nucleotide distance at the 3rd nucleotide position of codons and codon usage optimization to the human respiratory tract when BCoV and HCoV-OC43 were compared. An MCC tree based on relative synonymous codon usage (RSCU) data integrating data from both viruses and hosts into a same analysis indicated three putative host/virus contact dates ranging from 1.54E8 to 2.44E5 years ago, suggesting that an ancestor coronavirus might have followed human evolution.

Background Tuberculosis caused by Mycobacterium bovis is an important worldwide zoonosis and has been reported to cause clinical disease in several animal species, including captive wildlife. This report describes a case of M. bovis infection in a European bison from a Brazilian zoo and compiles a number of literature reports that raise concern regarding tuberculosis among captive wildlife in Brazil. Case presentation A 13 year-old captive-born male bison (Bison bonasus) from a Brazilian zoo began presenting weight loss, diarrhea and respiratory symptoms, which inevitably led to his death. At the animal’s necropsy, inspection of the thoracic and abdominal cavities revealed multiple enlarged lymph nodes, ranging from 4 to 10 cm, and pulmonary nodules containing caseous masses with firm white materials consistent with mineralization. Histopathology findings showed a significant amount of acid-alcohol resistant bacilli compatible with Mycobacterium spp. Specimens from lymph nodes and lungs were cultured on Petragnani and Stonebrink media, and specific PCR assays of the bacterial isolate identified it as M. bovis. Conclusion The European bison reported herein died from a severe form of disseminated tuberculosis caused by M. bovis. A review of the available literature indicates possible widespread occurrence of clinical disease caused by M. bovis or M. tuberculosis affecting multiple animal species in Brazilian wildlife-related institutions. These likely underestimated numbers raise concern regarding the control of the disease in captive animal populations from Brazil.

Highlights • We performed a search for canine coronavirus in fecal samples and organs from dogs. • Pantropic canine coronavirus was found in different organs. • Similar lesions to those found in canine parvovirus were observed. • This study reinforces the need for differential diagnosis between canine parvovirus and canine coronavirus., Characterization of canine coronavirus (CCoV) strains currently in circulation is essential for understanding viral evolution. The aim of this study was to determine the presence of pantropic CCoV type IIa in tissue samples from five puppies that died in Southern Brazil as a result of severe gastroenteritis. Reverse-transcriptase PCR was used to generate amplicons for sequence analysis. Phylogenetic analysis of the CCoV-IIa strains indicated that they were similar to those found in other countries, suggesting a common ancestor of these Brazilian isolates. This is the first report of pantropic CCoV-II in puppies from Latin America and our findings highlight that CCoV should be included as a differential diagnosis when dogs present with clinical signs and lesions typically seen with canine parvovirus infection.

Eastern equine encephalitis is a viral zoonosis that exhibits complex distribution and epidemiology, and greater importance should be given to this disease by the public-health authorities. In Brazil, although eastern equine encephalitis virus (EEEV) has been identified in vectors and antibodies are sometimes detected in horses and humans, there have been no records of equine encephalitis in horses caused by this virus during the last 24 years. This study describes eighteen cases of eastern equine encephalomyelitis that occurred in six Brazilian states between 2005 and 2009. Viral RNA was identified using semi-nested RT-PCR to detect members of the genus Alphavirus, and by genetic sequencing. The gene encoding NSP1 was partially amplified, and after genetic sequencing, eighteen sequences were generated. All eighteen strains were classified as belonging to lineage III of American EEEV. These findings could be an indication of the importance of this virus in animal and human public health.

Rotaviruses are a major cause of diarrhea in humans and animals, including several mammalian and avian species. Using different PCR protocols, we report the occurrence of rotavirus A in 21 (53.84%; 21/39) from 39 fecal pool samples of broilers, layers, and broiler breeders from Brazilian avian farms. We typed the G5, G8, G11, G19, and P[31] genotypes.

ObjectivesThe objectives of this study were to confirm the prevalence of feline immunodeficiency virus (FIV) infection in domestic cats in the region north of Ceará, Brazil, and to determine the factors associated with infection and the major circulating subtypes of the virus in this area.MethodsSamples from 148 cats were collected and tested using anti-FIV antibody screening, with confirmation of positive results by PCR. Univariate analysis was performed considering the epidemiological characteristics and FIV status. Sequencing and phylogenetic analysis of the gag and pol genes were performed to confirm the FIV subtype.ResultsNine cats (6.1%) tested positive for FIV – one female (0.7%) and eight males (5.4%). Male cats were significantly more likely to be infected ( P Conclusions and relevanceIn this study, we demonstrated a low prevalence for FIV in the northwest of Ceará, north-eastern Brazil. Male sex is a significant risk factor for FIV infection and the best predictive factor for FIV status. All isolates examined in this study clustered within subtype B, which is the predominant subtype in Brazil. This is the first report of genetic characterization of FIV in the state of Ceará, Brazil.

The application of viral metagenomic techniques and a series of PCRs in a human fecal sample enabled the detection of two novel circular unisense DNA viral genomes with 92% nucleotide similarity. The viruses were tentatively named circo-like virus-Brazil (CLV-BR) strains hs1 and hs2 and have genome lengths of 2526 and 2533 nucleotides, respectively. Four major open reading frames (ORFs) were identified in each of the genomes, and differences between the two genomes were primarily observed in ORF 2. Only ORF 3 showed significant amino acid similarities to a putative rolling circle replication initiator protein (Rep), although with low identity (36%). Our phylogenetic analysis, based on the Rep protein, demonstrated that the CLV-BRs do not cluster with members of the Circoviridae, Nanoviridae or Geminiviridae families and are more closely related to circo-like genomes previously identified in reclaimed water and feces of a wild rodent and of a bat. The CLV-BRs are members of a putative new family of circular Rep-encoding ssDNA viruses. Electron microscopy revealed icosahedral (~23 nm) structures, likely reflecting the novel viruses, and rod-shaped viral particles (~65-460 × 21 × 10 nm in length, diameter, and axial canal, respectively). Circo-like viruses have been detected in stool samples from humans and other mammals (bats, rodents, chimpanzees and bovines), cerebrospinal fluid and sera from humans, as well as samples from many other sources, e.g., insects, meat and the environment. Further studies are needed to classify all novel circular DNA viruses and elucidate their hosts, pathogenicity and evolutionary history.

With increasing frequency, avian Metapneumovirus (aMPV) is reported to induce respiratory signs in chickens. An adequate knowledge of current aMPV prevalence among Italian broilers is lacking, with little information available on its economical and health impact on the poultry industry. In order to collect preliminary data on the epidemiological context of aMPV in broiler flocks, a survey was performed in areas of Northern Italy with high poultry density from 2014 to 2016. Upper respiratory tract swabs were collected and processed by A and B subtype-specific multiplex real-time reverse transcription PCR (RT-PCR). Samples were also screened for infectious bronchitis virus (IBV) by generic RT-PCR and sequencing. Productive data and respiratory signs were detailed where possible. The high prevalence of aMPV was confirmed in broilers older than 26 d and also attested in IBV-negative farms. All aMPV detections belonged to subtype B. Italian strain genetic variability was evaluated by the partial attachment (G) gene sequencing of selected strains and compared with contemporary turkey strains and previously published aMPV references, revealing no host specificity and the progressive evolution of this virus in Italy.