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.

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.

Cross-sectional study of the G and P genotypes of rotavirus A field strains circulating in regularly vaccinated dairy cattle herds.

Neonatal diarrhea is the main cause of morbidity and mortality in calves up to 30 days old, and rotavirus A (RVA) is the main viral etiology. RVA vaccines are one of the main tools for diarrhea control in neonates. The aim of this cross-sectional study was to monitor by RT-PCR the G and P genotypes of RVA strains identified in dairy cattle herds regularly vaccinated with the RVA UK strain (G6P[5]). Of the 14 randomly selected herds, two were excluded because no calf was diagnosed with diarrhea on the day of fecal collection. Another six herds were also excluded from the study because all 20 diarrheic fecal samples evaluated were RT-PCR-negative. In the remaining six herds, 17 (25.4%) of the 67 diarrheic samples were RVA-positive. One G and P amplicon from each herd were selected for nucleotide sequencing. In the phylogenetic analysis, five RVA strains presented the G6P[11] genotype, and one presented the G10P[11] genotype. The G6 genotype present in all RVA field strains clustered into a distinct phylogenetic arrangement (lineage III) of the UK vaccine strain (lineage IV), characterizing the emergence of a phylogenetically distant G6 strain. In addition, we observed the emergence of strains with G10 and P[11] genotypes characterizing failure in heterologous immune protection. These results show the epidemiological importance of constant monitoring of RVA strains in vaccinated cattle herds and the low frequencies of diarrhea and diagnosis of RVA suggest that a regular vaccination program reduces the frequency and severity of RVA diarrhea in suckling calves.

Dairy calf rearing unit and infectious diseases: diarrhea outbreak by bovine coronavirus as a model for the dispersion of pathogenic microorganisms.

Dairy calf rearing unit is a management system that is only recently being implemented by some milk producer's cooperatives in southern Brazil. However, aspects related to the health profile of the heifer calves that arrive in the rearing unit as well as about biosecurity practices and microbiological challenges have not yet been evaluated in this rearing system in a tropical country. Diarrhea is the main and most frequent consequence of enteric infections in newborn calves. This study, through some etiological and epidemiological characteristics of an outbreak of neonatal diarrhea, has the aim to alert to the possibility of pathogenic microorganism spread in a dairy heifer calf rearing unit. The diarrhea outbreak presented some non-regular characteristics observed in bovine coronavirus (BCoV) enteric infections in dairy calves. The spread of infection was extremely rapid (1 week); the attack rate (> 50%) was much higher than that observed in calves subjected to conventional rearing; and the age range (5 to 90 days) of the affected heifer calves was much broader than that often observed in the BCoV diarrhea worldwide. These unusual epidemiological characteristics observed in this BCoV diarrhea outbreak raise awareness of the health threat present in calf rearing units as well as of the easy and rapid viral spread in a population of young animals from different dairy herds and, therefore, with very distinct immunological status.

Phylogenetic analysis of VP1 and RdRP genes of Brazilian aichivirus B strains involved in a diarrhea outbreak in dairy calves.

Aichivirus B has been reported worldwide in calves and adult cattle with and without diarrhea. The aim of this study was to describe the molecular characteristics of the RdRP and VP1 genes of aichivirus B strains identified as the most frequent etiologic agent in a neonatal diarrhea outbreak in a high-production Brazilian dairy cattle herd. Preliminary laboratory analysis ruled out important enteropathogens (Cryptosporidium spp; Eimeria spp., E. coli F5, and bovine coronavirus). Fecal samples from diarrheic (n = 24) and asymptomatic (n = 5) calves up to 30 days old were collected for virological analysis. RT-PCR assays were performed for the detection of aichivirus B RdRP and VP1 genes and for rotavirus A VP7 and VP4 genes in fecal samples. Asymptomatic calves (control group) were negative for both viruses. Aichivirus B and rotavirus A G10P[11] genotypes were found in 54.2% (13/24) and 25% (6/24) of the diarrheic fecal samples, respectively. Aichivirus B was only identified (83.3%, 10/12) in calves up to two weeks old. Phylogenetic analysis based on the RdRP gene grouped the Brazilian strains in a new branch within the aichivirus B group. Comparative analysis of the nucleotide sequence of the VP1 gene of Brazilian and Chinese aichivirus B strains allowed the strains identified in this study to be classified in the putative lineage 1. This is the first description of a high rate of aichivirus B detection in a diarrhea outbreak in dairy calves, and the first phylogenetic study of the VP1 gene of aichivirus B wild-type strains performed in South America.

Extra-intestinal detection of canine kobuvirus in a puppy from Southern Brazil.

This study presents the pathological, immunohistochemical, and molecular findings associated with the extra-intestinal detection of canine kobuvirus (CaKV) in a 5-month-old Chihuahua puppy, that had a clinical history of bloody-tinged feces. Principal pathological findings were interstitial pneumonia, necrotizing bronchitis, and parvovirus-induced enteritis. Molecular diagnostic methods identified CaKV within the cerebellum, cerebrum, lung, tonsil, and liver. CaKV and rotavirus were not identified within the feces and intestine. Immunohistochemistry (IHC) assays detected antigens of CDV and CAdV-1 in the lungs. These results confirmed the extra-intestinal detection of CaKV in this puppy and represent the first extra-intestinal detection of CaKV in a dog.

Genetic heterogeneity of the VP6 gene and predominance of G6P[5] genotypes of Brazilian porcine rotavirus C field strains.

The aim of this study was to investigate the genetic diversity of the VP6, VP7, and VP4 genes of 15 Brazilian wild-type porcine RVC strains identified in diarrheic fecal samples. The VP6 gene analysis demonstrated heterogeneity between the 15 RVC strains, which clustered in three distinct genotypes (I1, I5, and I6). In the VP7 and VP4 gene analysis, the genotype combination G6P[4] was detected in only one strain (UEL-77), while G6P[5] was the most commonly (n = 14) detected in RVC strains identified in the Brazilian pig herds evaluated, indicating its probable predominance in this country, mainly in 2014.

Isolation of an unusual G26P[13] group A rotavirus strain from piglets with diarrhea in Brazil.

This report describes the identification of an unusual G26P[13] genotype combination in a porcine group A rotavirus (RVA) strain. In a retrospective study, the VP7 (G type) and VP4 (P type) genes of porcine RVA Brazilian field strains identified in two diarrheic suckling piglets were amplified by RT-PCR and subjected to sequencing. The sequence analysis revealed the G26P[13] RVA genotype in one strain (BRA381) and G26P[X] in the other (BRA382). This study presents evidence of porcine RVA G26 genotype circulating in a Brazilian pig herd.

Molecular detection of bovine coronavirus in a diarrhea outbreak in pasture-feeding Nellore steers in southern Brazil.

Worldwide diarrhea outbreaks in cattle herds are more frequently detected in calves being that diarrhea outbreaks in adult cattle are not common. Winter dysentery (WD) is a bovine coronavirus (BCoV) enteric infection that is more reported in Northern hemisphere. Seasonal outbreaks of WD in adult cattle occur mainly in dairy cows. WD has not been described in beef cattle herds of tropical countries. This study describes the molecular detection of BCoV in a diarrhea outbreak in beef cattle steers (Nellore) raised on pasture in Parana, southern Brazil. During the outbreak, the farm had about 600 fattening steers. Watery and bloody diarrhea unresponsive to systemic broad-spectrum antibiotic therapy reveals a morbidity rate of approximately 15 %. The BCoV N gene was identified in 42.9 % (6/14) of the diarrheic fecal samples evaluated by semi-nested polymerase chain reaction (SN-PCR) technique. Other enteric microorganisms occasionally identified in adult cattle and evaluated in this study such as bovine groups A, B, and C rotavirus, bovine viral diarrhea virus, bovine torovirus, aichivirus B, and Eimeria sp. were not identified in the fecal samples. To the best knowledge of the authors, this is the first description of the BCoV diagnosis in fecal samples collected in a diarrhea outbreak in adult beef cattle grazing in the grass in a tropical country.

Age distribution of porcine sapovirus asymptomatic infection and molecular evidence of genogroups GIII and GIX? circulation in distinct Brazilian pig production systems.

This study aimed to evaluate the natural infection by SaV in pigs of different categories of production cycle in an important Brazilian pig-producing region. Faecal samples (n = 169) of suckling, post-weaning, finisher and breeder pig categories were analysed. Animals were from five farrow-to-weaning and nine grower-to-finish commercial pig farms. The RT-PCR assay was performed targeting the partial RNA-dependent RNA polymerase (RdRp) gene of porcine SaV genome. The virus was detected in 23.7% (40/169) of faecal samples and in 10/14 (5/5 farrow-to-weaning; 5/9 grower-to-finish) of pig farms evaluated. Porcine SaV was most frequently (p < 0.05) detected in pigs at post-weaning than in grower-to-finish and breeder categories. The phylogenetic analysis revealed that the porcine SaV strains belong to the GIII and GIX? genogroups. This study showed that the porcine SaV GIII genogroup has spread in the pig herds and provides the first evidence of GIX? genogroup circulation in South America.

Phylogenetic analysis of a G6P[5] bovine rotavirus strain isolated in a neonatal diarrhea outbreak in a beef cattle herd vaccinated with G6P[1] and G10P[11] genotypes.

The aim of this study was to perform the molecular characterization of the eleven genes of a G6P[5] bovine group A rotavirus (RVA) strain detected in a diarrhea outbreak from a vaccinated beef cattle herd. The outbreak affected 80 % of calves between 15-30 days old. RVA was identified by RT-PCR in 12 (70.6 %) out of 17 diarrheic fecal samples evaluated. The rotavirus wild-type strain had the genotype constellation G6(IV)-P[5](IX)-I2c-R2-C2-M2-A3-N2-T6-E2e-H3a. This study confirms the importance of homotypic immunity against the bovine RVA P[5] genotype in neonatal diarrhea in cattle herds that are regularly vaccinated against rotaviruses.

VP6 gene diversity in 11 Brazilian strains of porcine group C rotavirus.

Porcine group C rotavirus (RVC) is recognised as an enteric pathogen in piglets worldwide. The VP6 gene of RVC is divided into seven I-genotypes. Genotypes I2 and I3 are found in human and bovine strains, respectively; the porcine strains are divided into the other five genotypes (I1, I4-I7). In this study, molecular analysis of nearly the full length of the VP6 gene was performed in 11 Brazilian wild-type porcine RVC strains identified in diarrhoeic faecal samples, which were collected from eight pig farms located in five Brazilian states from piglets of 1-4 weeks of age. The nucleotide sequences of the VP6 gene showed 82.9-100 % identity between the Brazilian strains, 84.9-93.1 % with the prototype Cowden strain, and 82.4-92.2 % with other porcine RVC strains. In the 11 diarrhoeic faecal samples analysed in this study, three distinct porcine RVC genotypes (I1, I5, and I6) were identified and none were predominant. The results presented in this study revealed a high nucleotide diversity of the VP6 gene in porcine RVC field strains circulating in Brazil, which highlights the importance of further epidemiological and molecular surveys worldwide.

Species H rotavirus detected in piglets with diarrhea, Brazil, 2012.

We determined nucleotide and deduced amino acid sequences of the rotavirus gene encoding viral protein 6 from 3 fecal samples collected from piglets with diarrhea in Brazil, 2012. The analyses showed that the porcine rotavirus strains in Brazil are closely related to the novel species H rotavirus.

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 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.

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 evaluation of enteric viruses in asymptomatic free-ranging non-human primates (Alouatta guariba clamitans, Alouatta caraya, Callithrix spp., Callithrix penicillata, and Leontopithecus caissara) in southern Brazil.

BACKGROUND: This study evaluated the presence of rotavirus groups A, B, and C (RV-A, RV-B, and RV-C), sapovirus (SaV), and norovirus (NoV) in asymptomatic non-human primates (NHP). METHODS: Fecal samples were collected from 19 recently captured (Red-howler, Alouatta guariba clamitans, n = 18; Howler, Alouatta caraya, n = 1) and 43 free-ranging NHP (Marmosets, Callithrix spp., Callithrix penicillata, n = 30; Black-faced lion tamarin, Leontopithecus caissara, n = 12, Red-howler, Alouatta guariba clamitans, n = 1) that were maintained in southern Brazil without manifestation of diarrhea. Screening was performed by a combination of silver-stained polyacrylamide gel electrophoresis (ss-PAGE) and RT-PCR analyses. RESULTS: All samples were negative for RV-A, RV-B, RV-C, SaV, and NoV by both assays. CONCLUSION: The negative results obtained might be due to the absence of clinical manifestations of disease in the population of NHP evaluated.

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.

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.