Respiratory illness in young and adult cattle caused by bovine viral diarrhea virus subgenotype 2b in singular and mixed bacterial infection in a BVDV-vaccinated dairy herd.

Bovine respiratory disease (BRD) is a common global health problem in dairy cattle. The definitive diagnosis of BRD is complex because its etiology involves several predisposing and determining factors. This report describes the etiology of a BRD outbreak in a dairy herd in the mesoregion of Central Eastern Paraná, which simultaneously affected young (calves and heifers) and adult (cows) Holstein-Friesian cattle. Nine biological samples, consisting of five lung samples from two cows and three suckling calves, and four nasal swab samples from heifers, were used for etiological diagnosis. The nucleic acids extracted from lung fragments and nasal swabs were subjected to PCR and RT-PCR assays for partial amplification of the genes of five viruses [bovine viral diarrhea virus (BVDV), bovine alphaherpesvirus 1 (BoAHV1), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus 3 (BPIV-3), and bovine coronavirus (BCoV)] and four bacteria (Mycoplasma bovis, Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni) involved in the etiology of BRD. All nine biological samples from the animals with BRD tested negative for BoAHV1, BRSV, BPIV-3, BCoV, and H. somni. Therefore, the involvement of these microorganisms in the etiology of BRD outbreak can be ruled out. It was possible to identify the presence of BVDV and M. bovis in singular and mixed infections of the lower respiratory tract in cattle. BVDV was also identified in two nasal swabs: one as a single etiological agent and the other in association with two bacteria (P. multocida and M. haemolytica). The phylogenetic analysis conducted in the nucleotide sequence of the 5'UTR region and Npro gene of the BVDV amplicons demonstrated that the BVDV field strains of this BRD outbreak belong to subgenotype 2b. To the best of our knowledge, this is the first report of BVDV-2b involvement in the etiology of BRD in Brazil. Finally, it is necessary to highlight that the cattle were obtained from an open dairy herd with biannual vaccinations for BVDV-1a and - 2a.

Serological Profile for Major Respiratory Viruses in Unvaccinated Cows from High-Yielding Dairy Herds.

This study aims to determine the serological profile of high-yielding dairy cows for four main viruses (bovine alphaherpesvirus 1 (BoAHV1), bovine viral diarrhea virus (BVDV), bovine parainfluenza virus 3 (BPIV3), and bovine respiratory syncytial virus (BRSV)) related to bovine respiratory disease (BRD) in cattle herds worldwide. In this survey, 497 blood serum samples were collected from non-vaccinated dairy cows without clinical respiratory signs in 39 herds in the central-eastern mesoregion of Paraná State, South Brazil. The presence of neutralizing antibodies was determined by virus neutralization (VN) tests. VN antibodies against BoAHV1, BVDV, BPIV3, and BRSV were detected in 355 (71.4%), 280 (56.3%), 481 (96.8%), and 315 (63.4%) serum samples, respectively. The frequencies of seropositive herds for BoAHV1, BVDV, BPIV3, and BRSV were 79.5 (n = 31), 82.0 (n = 32), 100 (n = 39), and 84.6% (n = 33), respectively. The frequencies of seropositive cows varied according to the type of herd management and the number of cows in the herd. The detection of VN antibodies in unvaccinated dairy cattle herds demonstrated the endemic circulation of the four viruses in the herds evaluated. For BRD prevention, it is recommended to implement a vaccination program for cows that provides passive immunity in calves and active immunity in cows.

Outbreak of persistently infected heifer calves with bovine viral diarrhea virus subgenotypes 1b and 1d in a BVDV-vaccinated open dairy herd.

Bovine viral diarrhea virus (BVDV) infection has a significant economic impact on beef and dairy industries worldwide. Fetal infection with a non-cytopathic strain may lead to the birth of persistently infected (PI) offspring, which is the main event in the epidemiological chain of BVDV infection. This report describes the birth of 99 BVDV-PI heifer calves within 52 days of birth in a regular BVDV-vaccinated Brazilian dairy cattle herd and the subgenotypes of the infecting field strains. This study was conducted in a high-yielding open dairy cattle herd that frequently acquired heifers from neighboring areas for replacement. The farm monitors the birth of PI calves by screening all calves born using an ELISA (IDEXX) for BVDV antigen detection. All calves aged 1-7 days were evaluated. For positive and suspected results, the ELISA was repeated when the calves were close to one month old. A total of 294 heifer calves were evaluated between February and March 2021. Of these, 99 (33.7 %) had positive ELISA results and were considered PI calves. To evaluate the predominant BVDV species and subgenotypes in this outbreak, whole blood samples were collected from 31 calves born during the study period. All samples were submitted to the RT-PCR assay for the partial amplification of the BVDV 5'-UTR region, and these amplicons were subjected to nucleotide sequencing. Phylogenetic analysis identified BVDV-1b and BVDV-1d in 16 and 13 heifer calves, respectively. In two calves, it was not possible to determine the BVDV-1 subgenotype. Detection of PI animals and monitoring of circulating BVDV subgenotype strains are central to disease control. This study shows that regular BVDV vaccination alone may be insufficient to prevent BVDV infection in high-yielding open dairy cattle herds. Other biosecurity measures must be adopted to avoid the purchase of cattle with acute infections by BVDV or BVDV-PI, which can cause a break in the health profile of the herd and economic losses.

Serological Survey for Three Canine Viruses in Brazilian Wild Carnivores : Antibodies Against Canine Viruses in Wild Carnivores.

We evaluated the presence of antibodies against CaHV-1, CDV, and CPV-2 in serum samples from Brazilian wild carnivore species. Nine maned wolves and six crab-eating foxes were tested for CaHV-1 and CDV by virus neutralization test and CPV-2 by hemagglutination inhibition assay. Antibodies to CaHV-1, CDV, and CPV-2 were detected in serum samples of 1 (6.7%), 5 (33.3%), and 10 (66.7%) wild carnivores, respectively. Two maned wolves and one crab-eating fox were seropositive simultaneously for CDV and CPV-2. Antibodies against all viruses were detected in one crab-eating fox. This is the first report of CaHV-1 antibody detection in crab-eating foxes.

Serological profile of respiratory viruses in unvaccinated steers upon their arrival at Brazilian feedlot facilities.

Bovine viral diarrhea virus (BVDV), bovine alphaherpesvirus 1 (BoAHV1), bovine respiratory syncytial virus (BRSV), and bovine parainfluenza virus 3 (BPIV-3) are involved in bovine respiratory disease. These viruses can infect the respiratory system and cause considerable economic losses to beef and dairy cattle herds. This study aimed to determine the serological profiles of steers for BVDV, BoAHV1, BRSV, and BPIV-3 upon their arrival at Brazilian feedlot facilities. A total of 1,282 serum samples from unvaccinated steers were obtained on the first day of feeding. Samples were collected from 31 beef cattle herds reared in an extensive rearing system in six Brazilian states. Antibodies against BVDV, BoAHV1, BRSV, and BPIV-3 were detected using a virus neutralization test. The steers were distributed in agreement with their age and the Brazilian state of origin. The highest seropositivity was for BoAHV1 and BPIV-3 at 92.1% (1,154/1,253) and 86.6% (1,100/1,270), respectively. The seropositivity of BRSV was 77.1% (959/1,244). BVDV presented a lower rate, at slightly more than 50% (51.8%; 656/1,266). Age was a risk factor for the presence of antibodies against BVDV, BoAHV1, and BPIV-3 but not BRSV. A positive correlation was identified between BoAHV1 and BPIV-3 (P = 0.85) and between BRSV and BPIV-3 (P = 0.47). The high rate of seropositive steers for these four respiratory viruses on the first day of confinement identified in this serological survey provides important epidemiological information on respiratory infections, as the seropositivity of the four main bovine respiratory viruses in Brazilian beef cattle herds in an extensive rearing system.

The COVID-19 pandemic's impact on the practices and biosecurity measures of veterinary medicine professionals in Brazil.

Veterinarians, among other health professionals, are considered health professionals at high risk of exposure to and contraction of COVID-19. The main objective of this study is to assess changes in the clinical practices of veterinarians during the COVID-19 pandemic around prophylactic and biosafety measures, as well as to evaluate changes in workload and cost-benefit ratio. An online questionnaire was sent to veterinary professionals from July 2020 to July 2021 using Google Forms. A total of 1134 veterinarians answered the questionnaire on clinical experiences and biosafety practices during the COVID-19 pandemic. Veterinarians changed their routine clinical practices, as there was a reduction in working hours, and applied new patient approaches and advice to their owners, as well as restricting the number of people allowed inside. Biosafety measures were added in their workplaces, with an increase in the use of personal protective equipment. COVID-19 tests were administered at least once in 19.0%, and more than once in 9.5% of the respondents. Flu symptoms were present in 23.8% of the respondents, and 31.0% of the veterinarians attended to COVID-19 positive pet owners. Therefore, most veterinarians altered their routine practices, and some were exposed to sources of COVID-19 infection.

Os médicos veterinários, entre outros profissionais de saúde, são considerados profissionais de saúde sob alto risco de exposição e contração do COVID-19. O objetivo principal do presente estudo foi avaliar as mudanças na prática clínica de médicos veterinários durante a pandemia de COVID-19 em torno das medidas profiláticas e de biossegurança, assim como avaliar as mudanças na carga de trabalho e a relação custo-benefício. De julho a dezembro de 2020, um questionário online foi enviado aos profissionais por meio da ferramenta Formulários Google. Um total de 1.134 veterinários responderam ao questionário relacionado às experiências clínicas e práticas de biossegurança durante a pandemia COVID-19. Os médicos veterinários mudaram suas práticas clínicas rotineiras, pois houve redução da jornada de trabalho, novas abordagens dos pacientes e orientações aos proprietários, além da restrição do número de pessoas nos locais. Medidas de biossegurança foram adicionadas aos locais de trabalho, com aumento do uso de equipamentos de proteção individual. Os testes COVID-19 foram realizados pelo menos uma vez em 19,0% e mais de uma vez em 9,5% dos entrevistados. Sintomas de gripe estavam presentes em 23,8% dos profissionais entrevistados e 31,0% dos veterinários atendiam tratadores de animais positivos para COVID-19. Portanto, a maioria dos veterinários alterou suas práticas de rotina e alguns foram expostos a fontes de infecção COVID-19.

Congenital Neurological Disease Associated With HoBi-like Pestivirus Infection in a Newborn Dairy Calf From Brazil.

HoBi-like pestivirus (HoBiPeV) has been reported in several biological samples from cattle worldwide, but there are no descriptions of this virus associated with neurological symptoms. This report described the first occurrence of neurological disease associated with HoBiPeV in a newborn dairy calf. A mixed-breed Holstein calf had severe neurological symptoms at birth and died at 21 days old. The tissue fragments (central nervous system (CNS), myocardium, liver, kidney, lung, intestine, and spleen) were submitted to reverse transcription (RT)-PCR assay for the partial 5'-untranslated region (5'UTR) and N-terminal autoprotease (Npro) gene of the pestivirus genome, and the CNS tissue fragments were submitted to histopathological and immunohistochemical evaluation. The RT-PCR assay indicated that the kidney, CNS, and intestinal tissue fragments were positive for the pestivirus 5'UTR, and the CNS and intestinal tissue fragments were positive for the pestivirus Npro gene. Amplicons with high DNA quantification in the 5'UTR (CNS-cerebral cortex) and Npro (CNS-cerebral cortex and intestine) RT-PCR assays were sequenced. The nucleotide (nt) sequence and phylogenetic analysis of the 5'UTR strain exhibited 93.6 to 99.4%, 85%, 89.4 to 89.9%, 85.1%, and 90.5 to 91.5% nt identity with HoBiPeV strains from clades a, b, c, d, and e, respectively. The Npro amplicons showed 99.7% nt identity to each other and 90.4 to 96.5%, 85.1 to 85.3%, 79.2 to 79.7%, and 85.8 to 86.5% nt identity with HoBiPeV strains from clades a, c, d, and e, respectively. A histopathology revealed neuronal necrosis at the cerebrum, cerebellum, and brain stem. An immunohistochemical assay designed to identify antigens of bovine viral diarrhea virus revealed positive intracytoplasmic immunoreactivity within neurons at the cerebral cortex, cerebrum, cerebellum, and spinal cord. Thus, this report provides information about the first identification of HoBiPeV in tissues of the CNS in a newborn dairy calf with neurological symptoms.

Bacterial and fungal pathogens in granulomatous lesions of Chelonia mydas in a significant foraging ground off southern Brazil.

The green sea turtle Chelonia mydas inhabit near-shore areas exposed to threatening anthropogenic activities. The granulomatous lesions in these animals may indicate infectious diseases that can be associated with environmental contamination and hazards to human health. This study aimed to characterize the granulomatous inflammation associated with bacterial and fungal infection in C. mydas off Paraná state. From September 2015 to February 2019, systematic monitoring was performed by the Santos Basin Beach Monitoring Project for sea turtles'carcasses recovery, necropsy, and cause of death diagnosis. The tissue samples were fixed in buffered formalin 10% for histochemical analysis and frozen for molecular analysis to fungi detection (Internal Transcribed Spacer region of the nuclear rDNA) and bacteria detection (16S ribosomal gene). From a total of 270 C. mydas, granulomatous lesions were observed in different organs of 63 (23.3%) individuals. The histological analysis indicated lesions in 94 organs, affecting most respiratory and digestive systems. Bacteria were identified in 25 animals, including an acid-fast bacteria detected in one animal, and fungi in 24 C. mydas. The fungi species included the genus Candida (Candida zeylanoides, n = 3), Yarrowia (Yarrowia lipolytica, n = 9; Yarrowia deformans, n = 5; and Yarrowia divulgata, n = 1), and Cladosporium anthropophilum (n = 1). No species of bacteria was identified by molecular testing. All fungi species identified are saprobic, some are important to food and medical industries, but are also pathogens of humans and other animals. Therefore, long-term monitoring of these pathogens and the C. mydas health may indicate changes in environmental quality, possible zoonotic diseases, and their effects.

Investigation of avian rotavirus infections in broiler chicks from commercial flocks with different performance efficiency indexes.

The aim of this study was to investigate and compare the frequency of occurrence of avian rotavirus (AvRV) in poultry flocks according to its Performance Efficiency Index (PEI) scores. A total of 256 individual intestinal content samples of small sized-chicks (runts) with clinical signs of Runting Stunting Syndrome (RSS) and 24 clinically healthy chicks (control) were collected from twelve flocks in southern Brazil with different PEI scores: good (n = 4, PEI mean = 365); moderate (n = 4, PEI mean = 342) or poor (n = 4, PEI mean = 319). Silver-stained polyacrylamide gel electrophoresis (ss-PAGE) was used to detect and identify the AvRV species followed by RT-PCR and sequencing of the partial VP6 gene for species confirmation. AvRV was detected in 83% (10/12) of the flocks and 23.4% (60/256) of the chicks. The electrophoretic migration patterns of viral dsRNA segments were compatible with AvRV species A (AvRV- A), D (AvRV-D) and F (AvRV-F) in 9 (15%), 18 (30%), and 33 (55%) of the positive chicks fecal samples, respectively. The AvRV species identified by ss-PAGE were confirmed by RT-PCR and partial sequence analysis of the VP6 gene. The AvRV detection rate was statistically higher (p = 0.007) in chicks from flocks with poor PEI when compared to those with good PEI. The occurrence of AvRV-D and AvRV-F was statistically higher in 7 to 9 days old chicks, while AvRV-A was detected only in 13 to 14 days old animals.

First detection of Feline morbillivirus infection in white-eared opossums (Didelphis albiventris, Lund, 1840), a non-feline host.

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.

Severe outbreak of bovine neonatal diarrhea in a dairy calf rearing unit with multifactorial etiology.

This study describes the etiological diversity observed in a severe neonatal diarrhea outbreak with morbidity and mortality rates of 80 and 20%, respectively, with detection of mixed infections with viral, bacterial, and protozoan disease agents in a dairy calf rearing unit. Diarrheic fecal samples were collected from eight 5 to 18 days of age calves and were submitted to the investigation of the presence of rotavirus A (RVA), bovine coronavirus (BCoV), bovine kobuvirus (BKV), bovine viral diarrhea virus 1 and 2 (BVDV-1 and BVDV-2), enteropathogenic Escherichia coli (ETEC), Salmonella sp., and Cryptosporidium spp. Fragments of the small intestine of one calf with diarrhea that spontaneously died were submitted for histopathological analyses. The most frequent infectious agent detected in diarrheic fecal samples was BKV (8/8-100%), followed by RVA (5/8-62.5%), BVDV (5/8-62.5%), Cryptosporidium parvum (5/8-62.5%), ETEC (4/8-50%), and Cryptosporidium ryanae (1/8-12.5%). These etiological agents were found in mixed infections with two or more pathogens per diarrheic fecal sample. The association of viral and protozoan pathogens was the most frequently identified (37.5%) in these samples, followed by viral and bacterial (25%); viral, bacterial, and protozoan (25%); and only viral agents (12.5%). BCoV and Salmonella sp. were not identified in the diarrheic fecal samples analyzed. Additionally, histopathology of the small intestine diagnosed chronic lymphocytic enteritis. In conclusion, in calf rearing units, the adoption and strict monitoring of health management practices are critical to the success of this calf creation system.

Bovine viral diarrhea virus subgenotype 1a in a mummified fetus from a Brazilian dairy cattle herd.

We describe the molecular analysis of a wild-type field strain of bovine viral diarrhea virus (BVDV) identified in a mummified fetus from a small Brazilian dairy cattle herd. Nucleic acids extracted from samples of the lung, liver, heart, spleen, and kidney were tested by PCR assays for bovine alphaherpesvirus 1, Neospora caninum, Leptospira spp., Histophilus somni, and Brucella abortus, a nested PCR assay for Mycoplasma bovigenitalium and Ureaplasma diversum, and a RT-PCR assay for BVDV. Amplicons were only obtained in the RT-PCR assay for the partial amplification of the BVDV 5'UTR (288 bp) in kidney and spleen samples and the Npro (438 bp) gene in the kidney sample. Nucleotide sequencing of the amplified products and phylogenetic analyses based on the 2 BVDV genomic regions enabled the BVDV strain to be classified as subgenotype 1a.

Renicolidae infection in Manx shearwater (Puffinus puffinus): is parasitism implicated on renal lesions?

Renicolid digeneans are frequently observed in the renal tubules and ureters of seabirds, such Puffinus puffinus, a migratory species distributed along the Brazilian coast. However, few studies have focused on the relationship between renicolid infection and health status in P. puffinus. Thus, the aim of this study was to describe (i) renal and systemic alterations, (ii) the renicolids and (iii) the biological aspects associated with the presence of renicolids in P. puffinus. Gross and histological assays were performed in 93 P. puffinus stranded on the Paraná coast, southern Brazil, and renicolids were submitted to morphological and molecular assays. A high prevalence of renicolids in P. puffinus (71/93) was observed. In the kidney, the main microscopic findings were lymphocytic interstitial infiltrate, ductal ectasia and tubular necrosis. The renal lesions were significantly associated with the parasite infection. The morphological (n = 84) and molecular analyses (n = 2) confirmed the species as Renicola sloanei (100% and 95.9% of nucleotide identity with R. sloanei strains from P. puffinus and from Spheniscus demersus, respectively). In both parasitized and non-parasitized animals, cardiac and skeletal muscle degeneration and necrosis were the most frequent systemic changes. Therefore, the results suggest renicolids being a possible cause for the demonstrated renal alterations. A contribution of this parasite to a decreased health status of Puffinus puffinus along their migratory route is possible.

Molecular characterization of Feline paramyxovirus and Feline morbillivirus in cats from Brazil.

This study is aimed at detecting Feline paramyxovirus (FPaV) and Feline morbillivirus (FeMV) in 35 urine samples from domestic cats, collected in 2019, with or without clinical signs of uropathies using a reverse transcription-polymerase chain reaction (RT-PCR) followed by semi-nested polymerase chain reaction (SN-PCR) assays to amplify a partial paramyxovirus L gene. Eight (22.9%) out of the 35 urine samples were positive for paramyxoviruses. Sequencing and phylogenetic analyses revealed that three samples were positive for FPaV, four samples were positive for FeMV, and it was not possible to determine which virus was present in one RT-SN-PCR positive urine sample. FPaV strains showed 100% nucleotide (nt) identity with each other and 97% nt identity with a Japanese 163 FPaV strain. The FeMV strains showed 85.9% nt identity with each other; three strains were similar to previously described Brazilian FeMV strains, and one strain clustered in a different branch of the phylogenetic tree together with the first described Chinese FeMV strain. This study provides the first description of FPaV strains in cats from Brazil and provides new information about the molecular characteristics of FPaV and FeMV strains circulating in domestic cats in Brazil.

First detection of canine parvovirus 2b DNA in a crab-eating fox pup (Cerdocyon thous, Linnaeus, 1766).

The crab-eating fox (Cerdocyon thous) is a small wild mammal present in all Brazilian biomes and in some countries of South America. This study aimed to verify the involvement of viral infectious agents in the death of a wild crab-eating fox pup (Cerdocyon thous) in Brazil. The Center for Medicine and Research of Wild Animals of the Universidade Estadual Paulista received a free-living crab-eating fox aged approximately 21 days and apparently healthy. After 13 days, the animal presented anorexia, diarrhea, fever, prostration, and neurological signs progressing to death with an inconclusive diagnosis. In a retrospective study, tissue fragments stored at - 80 °C were used to identify nucleic acids from major canine viruses, such as canine parvovirus-2 (CPV-2), canine adenovirus A types 1 and 2, canid alphaherpesvirus 1, and canine distemper virus. The amplified product with the expected length for CPV-2 was obtained from the heart fragment. After performing nucleotide (nt) sequencing of the amplicon, it was possible to demonstrate that the crab-eating fox strain exhibited high (99.8%) nt identity with the CPV-2b prototype (CPV-39 strain). Additionally, deduced amino acid (aa) sequence analysis showed the GAT codon for the aa Asp (D) at position 426 of the CPV-2 viral protein VP2, which characterizes the subtype 2b. To the best of the authors' knowledge, this report describes the first detection of CPV-2b DNA in tissue fragments from a crab-eating fox.

Microbial diversity involved in the etiology of a bovine respiratory disease outbreak in a dairy calf rearing unit.

The etiological agents involved in a bovine respiratory disease (BRD) outbreak were investigated in a dairy heifer calf rearing unit from southern Brazil. A battery of PCR assays was performed to detect the most common viruses and bacteria associated with BRD, such as bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine alphaherpesvirus 1 (BoHV-1), bovine coronavirus (BCoV), bovine parainfluenza virus 3 (BPIV-3), Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. Bronchoalveolar lavage fluid (BALF) samples were taken from 21 heifer calves (symptomatic n = 15; asymptomatic n = 6) that, during the occurrence of the BDR outbreak, were aged between 6 and 90 days. At least one microorganism was detected in 85.7 % (18/21) of the BALF samples. Mixed infections were more frequent (72.2 %) than single infections (27.7 %). The interactions between viruses and bacteria were the most common in coinfections (55.5 %). The frequencies of BRD agents were 38.1 % for BRSV, 28.6 % for BVDV, 33.3 % for BCoV, 42.85 % for P. multocida, 33.3 % for M. bovis, and 19 % for H. somni. BoHV-1, BPIV-3, and M. haemolytica were not identified in any of the 21 BALF samples. Considering that BALF and not nasal swabs were analyzed, these results demonstrate the etiological multiplicity that may be involved in BRD outbreaks in dairy calves.

Genotype constellation of a rotavirus A field strain with an uncommon G8P[11] genotype combination in a rotavirus-vaccinated dairy cattle herd.

In this report we describe the genotype constellation of a bovine rotavirus A (RVA) strain with an uncommon G8P[11] genotype combination. The RVA/Cow-wt/BRA/Y136/2017/G8P[11] strain was classified as G8-P[11]-I2-R5-C2-M2-A3-N2-T9-E2-H3. Phylogenetic analysis based on the VP7 gene showed that the Y136 strain and a human G8P[1] strain comprise a putative new (VII) lineage for the G8 genotype. In addition, two other genotypes, R5 (VP1) and T9 (NSP3), were identified in the constellation of Y136 that are rarely found in RVA strains of bovine origin. The immunological pressure caused by regular vaccination of cows might be responsible for the selection of heterologous RVA strains.

Canine parvovirus 2b in fecal samples of asymptomatic free-living South American coatis (Nasua nasua, Linnaeus, 1766).

Canine parvovirus type 2 (CPV-2) is classified into three subtypes (CPV-2a, CPV-2b, and CPV-2c) and is the main cause of enteritis and myocarditis in young domestic and wild animals. This study aimed to evaluate the presence of CPV-2 in the feces of asymptomatic free-living coatis from Garden Forest Reserve, Palmital city, SP, Brazil. Fecal samples from 21 coatis (both sexes, different ages, and different aspects of feces) were collected in August 2014 and March 2015. The nucleic acid extracted was submitted to a polymerase chain reaction (PCR) assay to amplify a fragment of the VP2 gene of CPV-2. Eight (38%) fecal samples were positive in the PCR assay and were confirmed by sequencing. The 7 nucleotide (nt) sequences analyzed showed 100% nt identity with the prototype strain of CPV-2b (CPV-39 strain). The analysis of the deduced amino acid (aa) sequence revealed the presence of the GAT codon (aa D-Asp) at position 426 of the VP2 viral protein (subtype 2b). This study describes for the first time the identification of CPV-2b in asymptomatic free-living coatis (Nasua nasua) and suggests that coatis are susceptible to Carnivore protoparvovirus 1 infection and are important as a reservoir and an asymptomatic carrier to other wild and domestic animal species.

Longitudinal study of rotavirus C VP6 genotype I6 in diarrheic piglets up to 1 week old.

The reports of rotavirus C (RVC) involvement in diarrhea outbreaks in newborn piglets have been increasing in recent years. This longitudinal study, conducted over a 37-day period, aimed to evaluate the frequency of RVC infection in piglets aged up to 7 days obtained from a pig herd with a previous diagnosis of RVC infection in this age group. Piglets from 50 different litters were monitored daily for the occurrence of diarrhea, and all litters were classified into the following categories: sow parity order (PO) 1 to 5; litter size (LS) ≤ 10 piglets and > 10 piglets; and piglet birth weight (BW) 1.2 to 1.3 kg and > 1.3 to 1.4 kg. Two hundred six diarrheic fecal samples were collected and classified according to the fecal consistency score (pasty, semiliquid, liquid). Ten fecal samples were collected from asymptomatic piglets (control group). Fecal samples were screened for rotavirus (RV) by silver stained-polyacrylamide gel electrophoresis (ss-PAGE), and samples with inconclusive and negative-ss-PAGE results were submitted to RVC VP6 gene amplification by RT-PCR. RVC was identified in 71 (34.5%) samples, in 1 (10%) sample of the control group, and in piglets from 33 (66%) litters. The electrophoretic profile of RV species A was identified in only two samples. Of the 72 RVC-positive samples, 51 (70.8%) presented semiliquid or liquid consistency. There was no significant difference in either group regarding the production parameters (PO, LS, BW) evaluated. An analysis of the whole VP6 gene of three RVC field strains collected on the first, fifteenth, and last day of the experiment enabled us to identify genotype I6. This report describes the first longitudinal study examining epidemiological aspects of RVC infection in newborn piglets.

Neonatal diarrhea and rotavirus A infection in beef and dairy calves, Brazil, 2006-2015 / Diarreia neonatal e infecção por rotavírus A em bezerros de corte e leite, Brasil, 2006-2015

Calf diarrhea causes substantial economic losses in the cattle industry worldwide. Bovine rotavirus A (RVA) is the main viral agent that leads to enteric infection and diarrhea outbreaks in calves throughout the world. The aim of this retrospective (2006-2015) study was to determine the frequency of RVA detection in diarrheic fecal samples from beef and dairy calves from the three main cattle-producing regions of Brazil. Diarrheic fecal samples (n=1,498) of 124 beef and 56 dairy cattle herds from the Midwest, South, and Southeast geographical regions of Brazil were evaluated using the silver-stained polyacrylamide gel electrophoresis (ss-PAGE) technique. RVA double stranded-RNA was identified by the ss-PAGE technique in 410 (27.4%) fecal samples. The frequency of positive samples found in beef calves (31.9%; 328/1,027) was higher than the frequency found in diarrheic fecal samples from dairy calves (17.4%; 82/471). RVA infection was identified in calves from the three Brazilian geographical regions analyzed. However, the frequency of positive diarrheic calves in the Midwest region (39.4%), predominantly beef calves, was higher than in the South (19.4%) and Southeast (17.6%) regions. The temporal distribution of RVA-infected calves evaluated by two five-year periods (2006-2010, 24.5%; 2011-2015, 28.8%) demonstrated a very similar frequency of RVA in both periods. Considering the wide regional and temporal scope of this study, it can be concluded that RVA remains an important etiology of neonatal diarrhea in calves of Brazilian cattle herds.(AU)

A diarreia neonatal ocasiona perdas econômicas importantes na pecuária bovina em todo o mundo. Rotavírus A (RVA) é o principal agente etiológico viral de infecções entéricas e surtos de diarreia em bezerros de rebanhos de corte e leite. O objetivo deste estudo retrospectivo (2006-2015) foi determinar a frequência de detecção de RVA em amostras de fezes diarreicas de bezerros de corte e leite das três principais regiões produtoras de bovinos do Brasil. Amostras de fezes diarreicas (n=1.498) de 124 rebanhos bovinos de corte e 56 rebanhos bovinos de leite das regiões Centro-Oeste, Sul e Sudeste do Brasil foram avaliadas utilizando a técnica de eletroforese em gel de poliacrilamida (EGPA). O genoma segmentado de RVA foi identificado pela técnica de EGPA em 410 (27,4%) amostras de fezes. A frequência de amostras positivas encontrada em bezerros de rebanhos de corte (31,9%; 328/1.027) foi maior que a frequência identificada em amostras de fezes diarreicas de bezerros de rebanhos leiteiros (17,4%; 82/471). A infecção por RVA foi identificada em bezerros das três regiões geográficas brasileiras analisadas. No entanto, a frequência de bezerros com diarreia positivos para RVA na região Centro-Oeste (39,4%), predominantemente de bezerros de rebanhos de corte, foi maior que nas regiões Sul (19,4%) e Sudeste (17,6%). A distribuição temporal dos bezerros infectados com RVA avaliados por dois períodos de cinco anos (2006-2010, 24,5%; 2011-2015, 28,8%) demonstrou uma frequência muito semelhante em ambos os períodos. Considerando a amplitude regional e temporal deste estudo, pode-se concluir que RVA continua sendo uma importante etiologia de diarreia neonatal em bezerros de rebanhos bovinos brasileiros.(AU)