RESUMO
Bovine coronavirus (BCoV) is one of the important causes of diarrhoea in cattle. The virus is responsible for the high fatality rate associated with acute diarrhoea in calves. Rapid and accurate tests need to be conducted to detect the virus and minimise economic losses associated with the disease. Nucleic acid-based detection assays including PCR is an accurate test for detecting pathogens. However, these tests need skilled personnel, time and expensive devices. In this study, we developed a novel assay for the detection of BCoV in clinical cases. This novel assay combined reverse transcription-recombinase polymerase amplification with CRISPR/Cas13 and conducted a rapid visualisation of cleavage activity using a Lateral Flow Device. A conserved sequence of the BCV M gene was used as a target gene and the assays were tested in terms of specificity, sensitivity and time consumption. The result showed the specificity of the assay as 100% with no false positives being detected. Ten copies of the input RNA were enough to detect the virus and perform the assay. It took up to forty minutes for reading the results. Conducted together, the assay should be used as a rapid test to clinically diagnose infectious pathogens including bovine coronavirus. However, the assay needed the RNA to be extracted from the clinical sample in order to detect the virus. Therefore, more studies are needed to optimise the assay to be able to detect the virus in the clinical sample without extracting the RNA.
Assuntos
Sistemas CRISPR-Cas , Doenças dos Bovinos , Coronavirus Bovino , Diarreia , Sensibilidade e Especificidade , Animais , Bovinos , Coronavirus Bovino/isolamento & purificação , Coronavirus Bovino/genética , Diarreia/veterinária , Diarreia/virologia , Diarreia/diagnóstico , Doenças dos Bovinos/virologia , Doenças dos Bovinos/diagnóstico , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Infecções por Coronavirus/diagnósticoRESUMO
IMPORTANCE: Although the role of bovine coronavirus (BCoV) in calf diarrhea and respiratory disorders is well documented, its contribution to neurological diseases is unclear. OBJECTIVE: This study conducted virological investigations of calves showing diarrhea and respiratory and neurological signs. METHODS: An outbreak of diarrhea, respiratory, and neurological disorders occurred among the 12 calves in July 2022 in Istanbul, Türkiye. Two of these calves exhibited neurological signs and died a few days after the appearance of symptoms. One of these calves was necropsied and analyzed using molecular and histopathological tests. RESULTS: BCoV RNA was detected in the brain, lung, spleen, liver, and intestine of the calf that had neurological signs by real-time reverse transcription polymerase chain reaction. Immunostaining was also observed in the intestine and brain. A 622 bp S1 gene product was noted on gel electrophoresis only in the brain. Phylogenetic analysis indicated that the BCoV detected in this study had a high proximity to the BCoV strain GIb with 99.19% nucleotide sequence homology to the strains detected in Poland, Israel, Türkiye, and France. No distinct genetic lineages were observed when the brain isolate was compared with the respiratory and enteric strains reported to GenBank. In addition, the highest identity (98,72%) was obtained with the HECV 4408 and L07748 strains of human coronaviruses. CONCLUSIONS AND RELEVANCE: The strain detected in a calf brain belongs to the GIb-European lineage and shares high sequence homology with BCoV strains detected in Europe and Israel. In addition, the similarity between the human coronaviruses (4408 and L07748) raises questions about the zoonotic potential of the strains detected in this study.
Assuntos
Encéfalo , Doenças dos Bovinos , Infecções por Coronavirus , Coronavirus Bovino , Filogenia , Animais , Bovinos , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Infecções por Coronavirus/patologia , Doenças dos Bovinos/virologia , Doenças dos Bovinos/patologia , Encéfalo/virologia , Encéfalo/patologia , Turquia/epidemiologia , Surtos de Doenças/veterinária , Pneumonia Viral/veterinária , Pneumonia Viral/virologia , Pneumonia Viral/patologia , Doenças do Sistema Nervoso/veterinária , Doenças do Sistema Nervoso/virologia , Doenças do Sistema Nervoso/patologiaRESUMO
Coronaviruses have been confirmed to infect a variety of species, but only one case of associated winter dysentery of European bison has been described. The study aimed to analyze the prevalence, and define the impact on the species conservation, the source of coronavirus infection, and the role of the European bison in the transmission of the pathogen in Poland. Molecular and serological screening was performed on 409 European bison from 6 free-ranging and 14 captive herds over the period of 6 years (2017-2023). Presence of coronavirus was confirmed in one nasal swab by pancoronavirus RT-PCR and in 3 nasal swab samples by bovine coronavirus (BCoV) specific real time RT-PCR. The detected virus showed high (> 98%) homology in both RdRp and Spike genes to BCoV strains characterised recently in Polish cattle and strains isolated from wild cervids in Italy. Antibodies specific to BCoV were found in 6.4% of tested samples, all originating from free-ranging animals. Seroprevalence was higher in adult animals over 5 years of age (p = 0.0015) and in females (p = 0.09). Our results suggest that European bison play only a limited role as reservoirs of bovine-like coronaviruses. Although the most probable source of infections in the European bison population in Poland is cattle, other wild ruminants could also be involved. In addition, the zoonotic potential of bovine coronaviruses is quite low.
Assuntos
Bison , Infecções por Coronavirus , Animais , Bison/virologia , Polônia/epidemiologia , Feminino , Masculino , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Prevalência , Coronavirus/genética , Coronavirus/classificação , Coronavirus/isolamento & purificação , Estudos Soroepidemiológicos , Bovinos , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , Filogenia , Anticorpos Antivirais/sangueRESUMO
Bovine coronavirus (BCoV) poses a threat to cattle health worldwide, contributing to both respiratory and enteric diseases. However, few contemporary strains have been isolated. In this study, 71 samples (10 nasal and 61 fecal) were collected from one farm in Ohio in 2021 and three farms in Georgia in 2023. They were screened by BCoV-specific real-time reverse transcription-PCR, and 15 BCoV-positive samples were identified. Among them, five BCoV strains from fecal samples were isolated using human rectal tumor-18 (HRT-18) cells. The genomic sequences of five strains were obtained. The phylogenetic analysis illustrated that these new strains clustered with US BCoVs that have been detected since the 1990s. Sequence analyses of the spike proteins of four pairs of BCoVs, with each pair originally collected from the respiratory and enteric sites of one animal, revealed the potential amino acid residue patterns, such as D1180 for all four enteric BCoVs and G1180 for three of four respiratory BCoVs. This project provides new BCoV isolates and sequences and underscores the genetic diversity of BcoVs, the unknown mechanisms of disease types, and the necessity of sustained surveillance and research for BCoVs.
Assuntos
Doenças dos Bovinos , Infecções por Coronavirus , Coronavirus Bovino , Fezes , Filogenia , Bovinos , Animais , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , Coronavirus Bovino/classificação , Fezes/virologia , Doenças dos Bovinos/virologia , Infecções por Coronavirus/virologia , Infecções por Coronavirus/veterinária , Genoma Viral , Glicoproteína da Espícula de Coronavírus/genética , Humanos , Variação Genética , OhioRESUMO
Obtaining the complete or near-complete genome sequence of pathogens is becoming increasingly crucial for epidemiology, virology, clinical science and practice. This study aimed to detect viruses and conduct genetic characterization of genomes using metagenomics in order to identify the viral agents responsible for a calf's diarrhoea. The findings showed that bovine coronavirus (BCoV) and bovine rotavirus (BRV) are the primary viral agents responsible for the calf's diarrhoea. The current study successfully obtained the first-ever near-complete genome sequence of a bovine coronavirus (BCoV) from Türkiye. The G+C content was 36.31% and the genetic analysis revealed that the Turkish BCoV strain is closely related to respiratory BCoV strains from France and Ireland, with high nucleotide sequence and amino acid identity and similarity. In the present study, analysis of the S protein of the Turkish BCoV strain revealed the presence of 13 amino acid insertions, one of which was found to be shared with the French respiratory BCoV. The study also identified a BRV strain through metagenomic analysis and detected multiple mutations within the structural and non-structural proteins of the BRV strain, suggesting that the BRV Kirikkale strain may serve as an ancestor for reassortants with interspecies transmission, especially involving rotaviruses that infect rabbits and giraffes.
Assuntos
Coronavirus Bovino , Genoma Viral , Metagenômica , Rotavirus , Animais , Metagenômica/métodos , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , Bovinos , Rotavirus/genética , Rotavirus/isolamento & purificação , Rotavirus/classificação , Turquia , Doenças dos Bovinos/virologia , Infecções por Rotavirus/veterinária , Infecções por Rotavirus/virologiaRESUMO
BACKGROUND: Bovine coronavirus (BCoV) is implicated in severe diarrhea in calves and contributes to the bovine respiratory disease complex; it shares a close relationship with human coronavirus. Similar to other coronaviruses, remarkable variability was found in the genome and biology of the BCoV. In 2022, samples of feces were collected from a cattle farm. A virus was isolated from 7-day-old newborn calves. In this study, we present the genetic characteristics of a new BCoV isolate. The complete genomic, spike protein, and nucleocapsid protein gene sequences of the BCoV strain, along with those of other coronaviruses, were obtained from the GenBank database. Genetic analysis was conducted using MEGA7.0 and the Neighbor-Joining (NJ) method. The reference strains' related genes were retrieved from GenBank for comparison and analysis using DNAMAN. RESULTS: The phylogenetic tree and whole genome consistency analysis showed that it belonged to the GIIb subgroup, which is epidemic in Asia and America, and was quite similar to the Chinese strains in the same cluster. Significantly, the S gene was highly consistent with QH1 (MH810151.1) isolated from yak. This suggests that the strain may have originated from interspecies transmission involving mutations of wild strains. The N gene was conserved and showed high sequence identity with the epidemic strains in China and the USA. CONCLUSIONS: Genetic characterization suggests that the isolated strain could be a new mutant from a wild-type lineage, which is in the same cluster as most Chinese epidemic strains but on a new branch.
Assuntos
Doenças dos Bovinos , Infecções por Coronavirus , Coronavirus Bovino , Genoma Viral , Filogenia , Animais , Bovinos , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , China/epidemiologia , Doenças dos Bovinos/virologia , Doenças dos Bovinos/epidemiologia , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Infecções por Coronavirus/epidemiologia , Fezes/virologia , Glicoproteína da Espícula de Coronavírus/genética , Animais Recém-NascidosRESUMO
In this study, virological surveillance focused on coronaviruses in marmots in the Alpine region in 2022, captured as part of a population control reduction program in the Livigno area. Seventy-six faecal samples were randomly collected from marmots at the time of capture and release and tested for genome detection of pan-coronavirus, pan-pestivirus, canine distemper virus, and influenza A and D virus. Nine faecal samples were positive in the Pan-CoV RT-PCR, while all were negative for the other viruses. Pan-coronavirus positives were further identified using Illumina's complete genome sequencing, which showed the highest homology with Bovine Coronavirus previously detected in roe deer in the Alps. Blood samples (n.35) were collected randomly from animals at release and tested for bovine coronavirus (BCoV) antibodies using competitive ELISA and VNT. Serological analyses revealed that 8/35 sera were positive for BCoV antibodies in both serological tests. This study provides molecular and serological evidence of the presence of BCoV in an alpine marmot population due to a likely spillover event. Marmots share areas and pastures with roe deer and other wild ruminants, and environmental transmission is a concrete possibility.
Assuntos
Anticorpos Antivirais , Coronavirus Bovino , Fezes , Marmota , Filogenia , Animais , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , Marmota/virologia , Fezes/virologia , Anticorpos Antivirais/sangue , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Infecções por Coronavirus/diagnóstico , Bovinos , Ensaio de Imunoadsorção Enzimática , Genoma ViralRESUMO
Bovine coronavirus (BCoV) has dual tropisms that can trigger enteric and respiratory diseases in cattle. Despite its global distribution, BCoV field strains from Brazil remain underexplored in studies investigating the virus's worldwide circulation. Another research gap involves the comparative analysis of S protein sequences in BCoV isolates from passages in cell lines versus direct sequencing from clinical samples. Therefore, one of the objectives of our study was to conduct a comprehensive phylogenetic analysis of BCoV strains identified from Brazil, including a respiratory strain obtained during this study, comparing them with global and ancestral BCoV strains. Additionally, we performed a comparative analysis between wild-type BCoV directly sequenced from the clinical sample (nasal secretion) and the cell culture-adapted strain, utilizing the Sanger method. The field strain and multiple cell passage in cell culture (HRT-18) adapted BCoV strain (BOV19 NS) detected in this study were characterized through molecular and phylogenetic analyses based on partial fragments of 1,448 nt covering the hypervariable region of the S gene. The analyses have demonstrated that different BCoV strains circulating in Brazil, and possibly Brazilian variants, constitute a new genotype (putative G15 genotype). Compared with the ancestral prototype (Mebus strain) of BCoV, 33 nt substitutions were identified of which 15 resulted in non-synonymous mutations (nine transitions and six transversions). Now, compared with the wild-type strain was identified only one nt substitution in nt 2,428 from the seventh passage onwards, which resulted in transversion, neutral-neutral charge, and one substitution of asparagine for tyrosine at aa residue 810 (N810Y).
Assuntos
Doenças dos Bovinos , Coronavirus Bovino , Filogenia , Bovinos , Brasil , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , Coronavirus Bovino/classificação , Animais , Doenças dos Bovinos/virologia , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Genótipo , Glicoproteína da Espícula de Coronavírus/genética , Infecções Respiratórias/virologia , Linhagem CelularRESUMO
Background: Diarrhea in newborn small ruminants continues to be the cause of significant financial loss in sheep and goat farms worldwide. Commercial immunochromatographic (IC) assays have been designed and evaluated to be used for the diagnosis of diarrhea in cattle; however, there are no trials to use rapid tests in small ruminants. Aim: This study was carried out in Kuwait to evaluate the performance of the rapid immunochromatography test (BoviD-4, BioNote, Inc, Korea) for diagnostics of Cryptosporidium, rotavirus A (RVA), bovine coronavirus (BCoV), and Escherichia coli K99 (E. coli K99) in fecal samples of sheep and goats. Methods: A total of 85 samples were examined using BoviD-4, and the results were compared with that of polymerase chain reaction for Cryptosporidium, RVA, and BCoV, whereas for E. coli K99 it was by isolation and identification as reference tests. Results: The kappa test agreement results between the BoviD-4 and reference tests were 0.870 (perfect), 0.783 (substantial), 0.728 (substantial), and 0.281 (fair) for the detection of E. coli K99, Cryptosporidium, RVA, and BCoV, respectively. The sensitivity of BoviD-4 kit was 91.2%, 80.0%, 90.0%, and 37.5% and the specificity was 88.2%, 96.0%, 96.4%, and 92.2% for Cryptosporidium, RVA, E. coli K99, and BCoV, respectively. Conclusion: The Bovid-4 kit can be used as a rapid pen-side test for Cryptosporidium spp., E. coli K99, and RVA in the field. Nonetheless, care must be taken while interpreting the BCoV results of the kit.
Assuntos
Cromatografia de Afinidade/veterinária , Coronavirus Bovino , Cryptosporidium , Escherichia coli , Rotavirus , Animais , Coronavirus Bovino/isolamento & purificação , Criptosporidiose/diagnóstico , Cryptosporidium/isolamento & purificação , Escherichia coli/isolamento & purificação , Fezes , Cabras , Kuweit , Rotavirus/isolamento & purificação , OvinosRESUMO
BACKGROUND: Bovine rotavirus (BRV) and bovine coronavirus (BCoV) are the most common viral agents in neonatal calf diarrhea and result in serious economic consequences. The aim of the study was to determine the epidemiology of those viruses in randomly selected dairy farms of Addis Ababa. METHODS: A cross-sectional study was conducted from November 2018 to April 2019 using a probability proportional to size (PPS) sampling technique. A total of 110 calves, less than 30 days of age, from 57 dairy herds were involved in the study. Associated factors of herds and calves were collected using semistructured interviews from farm owners and through physical observation of selected calves. Fecal samples were collected and analyzed using the sandwich ELISA method. Data generated from both semistructured interviews and laboratory investigation were analyzed using STATA_MP version 15. RESULTS: From the total 110 calves, 42 (38.18%) had diarrhea during the survey. The prevalence of bovine rotavirus and coronavirus was 3.64% (4/110) and 0.91% (1/110), respectively. Diarrhea, feeding colostrum timing, and sex of the neonatal calves had statistically significant association with bovine rotavirus infection (p < 0.05). All rotavirus-positive neonatal calves were identified in small scale dairy farms and in dairy farms that reported mortality though they lack statistically significant association. Only one coronavirus case was detected among the neonatal calves. The case was identified among small scale herds and in a herd with diarrheal cases. The sex of the coronavirus calf was female, diarrheic, and among 11-20 days old. CONCLUSION: The prevalence of rotavirus and coronavirus infections in neonatal calves was seldom in dairy farms of the study area. Rotavirus was more common than coronavirus, and further studies should be initiated on other (infectious and noninfectious) causes of neonatal calf diarrhea in the area.
Assuntos
Doenças dos Bovinos/epidemiologia , Infecções por Coronavirus/veterinária , Infecções por Rotavirus/veterinária , Animais , Animais Recém-Nascidos , Bovinos , Doenças dos Bovinos/virologia , Infecções por Coronavirus/epidemiologia , Coronavirus Bovino/isolamento & purificação , Estudos Transversais , Diarreia/epidemiologia , Diarreia/veterinária , Diarreia/virologia , Etiópia/epidemiologia , Fazendas/estatística & dados numéricos , Fezes/virologia , Feminino , Masculino , Mortalidade , Prevalência , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologiaRESUMO
Bovine coronavirus (BCoV) is the causative agent of winter dysentery (WD). In adult dairy cattle, WD is characterized by hemorrhagic diarrhea and a reduction in milk production. Therefore, WD leads to significant economic losses in dairy farms. In this study, we aimed to isolate and characterize local BCoV strains. BCoV positive samples, collected during 2017-2021, were used to amplify and sequence the S1 domain of S glycoprotein and the full hemagglutinin esterase gene. Based on our molecular analysis, local strains belong to different genetic variants circulating in dairy farms in Israel. Phylogenetic analysis revealed that all local strains clustered together and in proximity to other BCoV circulating in the area. Additionally, we found that local strains are genetically distant from the reference enteric strain Mebus. To our knowledge, this is the first report providing molecular data on BCoV circulating in Israel.
Assuntos
Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/genética , Disenteria/veterinária , Filogenia , Animais , Antígenos Virais/genética , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Coronavirus Bovino/classificação , Coronavirus Bovino/isolamento & purificação , Indústria de Laticínios , Disenteria/virologia , Fezes/virologia , Feminino , Variação Genética , Israel/epidemiologia , Análise de Sequência de DNARESUMO
Calf gastrointestinal disease remains one of the main causes of productivity and economic losses on dairy operations. The majority of pre-weaned calf mortality is attributed to diarrhea or other digestive problems. Five enteric pathogens are commonly associated with diarrhea in dairy calves, including bovine rotavirus, bovine coronavirus, Escherichia coli, Salmonella spp., and Cryptosporidium parvum. Pathogen-associated differences in health outcomes and case fatality rates have not been well-characterized. Additionally, updated prevalence estimates may reflect important changes in the epidemiology of the pathogens on dairy farms. For this cohort study, fecal samples were collected from 276 clinically ill calves across 5 central Ohio dairy farms on the first day of diarrheal diagnosis. Genomic techniques, including reverse transcription polymerase chain reaction (RT-PCR) and droplet digital polymerase chain reaction (ddPCR) were used to test for the presence of the five enteric pathogens. A Poisson regression model was used to estimate the relative risk of mortality, and a survival analysis with a Cox regression model was used to analyze time to return to a healthy clinical status by pathogen. Rotavirus was the most frequently identified at 68.1 % (188/276), followed by F5 (K99)+E. coli at 42.5 % (114/268), C. parvum at 28.4 % (66/232), coronavirus at 5.8 % (16/276), and Salmonella had the lowest prevalence at 3.7 % (10/268). Risk of mortality tended to be higher for calves infected with Salmonella (RR = 3.83; 95 %CI: 0.93, 16.02, p = 0.062); however, the time to return to a healthy clinical status was not different for different pathogens. Only farm was a significant predictor of time to return to health (p = 0.017); the within-farm median duration of signs substantially varied between 2 and 7 days. The results suggest that the prevalence and distribution of rotaviral infections is higher than reported in prior studies. With the exception of infections caused by Salmonella spp., pathogen diagnosis on the first day of diarrhea was a poor predictor of the outcome and duration of disease. These results are critical to guide the implementation of prevention measures to detect, treat, and prevent calf diarrhea.
Assuntos
Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/virologia , Diarreia/veterinária , Animais , Animais Recém-Nascidos , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/prevenção & controle , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/isolamento & purificação , Criptosporidiose/epidemiologia , Cryptosporidium parvum/isolamento & purificação , Indústria de Laticínios , Diarreia/epidemiologia , Diarreia/microbiologia , Diarreia/virologia , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/veterinária , Fazendas , Estudos Longitudinais , Ohio/epidemiologia , Prevalência , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/veterinária , Salmonella/isolamento & purificação , Salmonelose Animal/epidemiologia , Resultado do Tratamento , DesmameRESUMO
BRD is associated with infectious agents, but management and transport-stress are trigger factors. Metaphylactic administration of antimicrobial reduces colonization of respiratory tract by pathogens, but the development of antibiotic-resistance raises public health concerns leading to propose new control strategies. The study analyzed nasopharyngeal swabs of 231 imported cattle, 10% of 49 trucks, transported from France to southern Italy and, through Real-time PCR identified the prevalence of the involved pathogens speculating on strategies to reduce the impact of BRD. The samples were tested by Real-time PCR, for the detection of bovine coronavirus (BCoV), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus (BPiV), bovine adenovirus (BAdV), Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. Yates-corrected chi squared, or Fisher's exact test were used to compare both animal-health status and positivity/negativity to pathogens, and the relationship between presence/absence of clinical signs and Real-time PCR-positivity. H. somni and BCoV were the most frequently identified pathogens. In BRD-diagnosed cattle, BAdV was detected in 13.8% (19/138), BRSV in 14.5% (20/138) and BPiV in 4.3% (6/138). Healthy cattle were mostly positive for H. somni (89.2%, 83/93). A statistically significant association was observed between clinical signs and positivity to M. haemolytica (p value = 0.016). Although mass-medication and vaccination are used for BRD control, it still remains a primary health problem. Our results highlight that the nasopharyngeal microbiota could be affected by transport and that strategies to enhance calf immunity for reducing BRD-risk development would be more effective if applied at farm of origin prior to loading.
Assuntos
Doenças dos Bovinos/epidemiologia , Coronavirus Bovino/isolamento & purificação , Microbiota , Pasteurellaceae/isolamento & purificação , Doenças Respiratórias/veterinária , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/prevenção & controle , Coronavirus Bovino/genética , Estudos Epidemiológicos , França/epidemiologia , Imunidade , Itália/epidemiologia , Masculino , Mastadenovirus/genética , Mastadenovirus/isolamento & purificação , Nasofaringe/microbiologia , Pasteurellaceae/genética , Vírus Sincicial Respiratório Bovino/genética , Vírus Sincicial Respiratório Bovino/isolamento & purificação , Sistema Respiratório/microbiologia , Doenças Respiratórias/epidemiologia , Doenças Respiratórias/microbiologia , Doenças Respiratórias/prevenção & controle , Respirovirus/genética , Respirovirus/isolamento & purificação , Meios de TransporteRESUMO
BACKGROUND: Apart from the huge worldwide economic losses often occasioned by bovine coronavirus (BCoV) to the livestock industry, particularly with respect to cattle rearing, continuous surveillance of the virus in cattle and small ruminants is essential in monitoring variations in the virus that could enhance host switching. In this study, we collected rectal swabs from a total of 1,498 cattle, sheep and goats. BCoV detection was based on reverse transcriptase polymerase chain reaction. Sanger sequencing of the partial RNA-dependent RNA polymerase (RdRp) region for postive samples were done and nucleotide sequences were compared with homologous sequences from the GenBank. RESULTS: The study reports a BCoV prevalence of 0.3%, consisting of 4 positive cases; 3 goats and 1 cattle. Less than 10% of all the animals sampled showed clinical signs such as diarrhea and respiratory distress except for high temperature which occurred in > 1000 of the animals. However, none of the 4 BCoV positive animals manifested any clinical signs of the infection at the time of sample collection. Bayesian majority-rule cladogram comparing partial and full length BCoV RdRp genes obtained in the study to data from the GenBank revealed that the sequences obtained from this study formed one large monophyletic group with those from different species and countries. The goat sequences were similar to each other and clustered within the same clade. No major variations were thus observed between our isolates and those from elsewhere. CONCLUSIONS: Given that Ghana predominantly practices the extensive and semi-intensive systems of animal rearing, our study highlights the potential for spillover of BCoV to small ruminants in settings with mixed husbandry and limited separation between species.
Assuntos
Doenças dos Bovinos/virologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/isolamento & purificação , Doenças das Cabras/virologia , Doenças dos Ovinos/virologia , Animais , Sequência de Bases , Teorema de Bayes , Bovinos , Doenças dos Bovinos/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Coronavirus Bovino/genética , Diarreia/veterinária , Gana/epidemiologia , Doenças das Cabras/epidemiologia , Cabras , Filogenia , Prevalência , RNA Polimerase Dependente de RNA/genética , Síndrome do Desconforto Respiratório/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Ovinos , Doenças dos Ovinos/epidemiologiaRESUMO
The hemagglutinin-esterase (HE) protein of betacoronavirus lineage A is a secondary receptor in the infection process and is involved in the emergence of new betacoronavirus genotypes with altered host specificity and tissue tropism. We previously reported a novel recombinant bovine coronavirus (BCoV) strain that was circulating in dairy cattle in China, but this virus was not successfully isolated, and the genetic characteristics of BCoV are still largely unknown. In this study, 20 diarrheic faecal samples were collected from a farm in Liaoning province that had an outbreak of calf diarrhea (≤ 3 months of age) in November 2018, and all of the samples tested positive for BCoV by RT-PCR. In addition, a BCoV strain with a recombinant HE (designated as SWUN/A1/2018) and another BCoV strain with a recombinant HE containing an insertion (designated as SWUN/A10/2018) were successfully isolated in cell culture (TCID50: 104.25/mL and 104.73/mL, respectively). Unexpectedly, we identified the emergence of a novel BCoV variant characterized by a 12-nt bovine gene insertion in the receptor-binding domain in a natural recombinant HE gene, suggesting a novel evolutionary pattern in BCoV.
Assuntos
Doenças dos Bovinos/epidemiologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/genética , Diarreia/veterinária , Hemaglutininas Virais/genética , RNA Viral/genética , Proteínas Virais de Fusão/genética , Animais , Bovinos , Doenças dos Bovinos/patologia , Doenças dos Bovinos/virologia , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Coronavirus Bovino/classificação , Coronavirus Bovino/isolamento & purificação , Diarreia/epidemiologia , Diarreia/patologia , Diarreia/virologia , Evolução Molecular , Fezes/virologia , Expressão Gênica , Genótipo , Modelos Moleculares , Mutagênese Insercional , Filogenia , Estrutura Secundária de Proteína , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
Bovine respiratory disease complex is etiologically complex and usually involves co-infection by several agents, including bovine parainfluenza virus-3 (BPIV-3), bovine respiratory syncytial virus (BRSV), and bovine coronavirus (BCoV). Traditionally, vaccines have been tested in seronegative calves infected with a single in vitro-passaged agent, often with little disease, resulting in unvaccinated subjects. To overcome the potential problem of attenuation coincident with in vitro culture of the viruses, cocktails of field isolates of BPIV-3s and BCoVs were passaged in the lungs of neonatal colostrum-deprived calves. Lung lavage fluids were used as inocula, alone and in combination with in-vivo passaged BRSV, and aerosolized into a trailer containing conventionally reared 9-week-old weaned Holstein calves with decayed, but still measurable, maternal antibodies. Calves developed acute respiratory disease of variable severity. Upon necropsy, there were characteristic gross and histologic lesions in the respiratory tract, associated immunohistochemically with BPIV-3, BRSV, and BCoV. In-vivo passage of viruses is an alternative to in vitro culture to produce inocula to better study the pathogenesis of infection and more rigorously and relevantly assess vaccine efficacy.
Le complexe des maladies respiratoires bovines possède une étiologie complexe et implique habituellement une co-infection par plusieurs agents, incluant le virus parainfluenza bovin 3 (BPIV-3), le virus respiratoire syncitial bovin (BRSV) et le coronavirus bovin (BCoV). Traditionnellement, les vaccins ont été testés chez des veaux séronégatifs infectés avec un seul agent cultivé in vitro, présentant souvent peu de maladie, résultant en des sujets non-vaccinés. Afin de contrecarrer le problème potentiel d'atténuation associé à la culture in vitro des virus, des cocktails d'isolats de champs de BPIV-3 et de BCoV furent passés dans des poumons de veaux nouveau-nés privés de colostrum. Les liquides de lavage pulmonaire furent utilisés comme inoculum, seul et en combinaison avec des BRSV passés in vivo, et aérosolisés dans une remorque contenant des veaux Holstein sevrés élevés de manière conventionnelle âgés de 9 semaines ayant des anticorps maternels en déclin mais toujours mesurables. Les veaux ont développé une maladie respiratoire aiguë de sévérité variable. Lors de la nécropsie, il y avait des lésions macroscopiques et histologiques caractéristiques dans le tractus respiratoire, associées immuno-histochimiquement avec BPIV-3, BRSV et BCoV. Le passage in vivo de virus est une alternative à la culture in vitro afin de produire un inoculum permettant de mieux étudier la pathogénie de l'infection et d'évaluer plus rigoureusement et plus pertinemment l'efficacité de vaccins.(Traduit par Docteur Serge Messier).
Assuntos
Doenças dos Bovinos/virologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/patogenicidade , Vírus da Parainfluenza 3 Bovina/patogenicidade , Infecções por Vírus Respiratório Sincicial/veterinária , Infecções por Respirovirus/veterinária , Animais , Anticorpos Antivirais/sangue , Antígenos Virais/isolamento & purificação , Bovinos , Doenças dos Bovinos/patologia , Infecções por Coronavirus/complicações , Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Coronavirus Bovino/isolamento & purificação , Imuno-Histoquímica/veterinária , Pulmão/patologia , Pulmão/virologia , Vírus da Parainfluenza 3 Bovina/imunologia , Vírus da Parainfluenza 3 Bovina/isolamento & purificação , Atelectasia Pulmonar/patologia , Atelectasia Pulmonar/veterinária , Infecções por Vírus Respiratório Sincicial/complicações , Infecções por Vírus Respiratório Sincicial/patologia , Vírus Sinciciais Respiratórios/isolamento & purificação , Vírus Sinciciais Respiratórios/patogenicidade , Infecções por Respirovirus/complicações , Infecções por Respirovirus/patologia , Infecções por Respirovirus/virologia , Traqueia/patologia , Traqueia/virologiaRESUMO
Bovine coronaviruses are spread all over the world. They cause two types of clinical manifestations in cattle either an enteric, calf diarrhoea and winter dysentery in adult cattle, or respiratory in all age groups of cattle. The role of coronaviruses in respiratory infections is still a hot topic of discussion since they have been isolated from sick as well as healthy animals and replication of disease is rarely successful. Bovine coronavirus infection is characterised by high morbidity but low mortality. The laboratory diagnosis is typically based on serological or molecular methods. There is no registered drug for the treatment of virus infections in cattle and we are limited to supportive therapy and preventative measures. The prevention of infection is based on vaccination, biosecurity, management and hygiene. This paper will cover epidemiology, taxonomy, pathogenesis, clinical signs, diagnosis, therapy, economic impact and prevention of coronavirus infections in cattle.
Assuntos
Doenças dos Bovinos/virologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/terapia , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/terapia , Coronavirus Bovino/classificação , Coronavirus Bovino/isolamento & purificação , Coronavirus Bovino/fisiologia , Diarreia/veterinária , Diarreia/virologia , Enterocolite Necrosante/veterinária , Enterocolite Necrosante/virologia , Infecções Respiratórias/veterinária , Infecções Respiratórias/virologia , Vacinação/veterináriaRESUMO
Group A rotavirus (RVA) and bovine coronavirus (BCoV) are the two main viral enteropathogens associated with neonatal calf diarrhea. The aim of the present survey was to investigate the epidemiology and the role of RVA and BCoV in the presentation of dairy and beef calf diarrhea in Lerma Valley of Salta province, within the Northwest region of Argentina. Stool samples of calves with or without diarrhea younger than 2 months of age were collected from 19 dairy farms and 20 beef farms between the years 2014 and 2016. Stool samples were screened for RVA and BCoV detection by ELISA. Heminested multiplex RT-PCR was used for RVA typing and RT-PCR to confirm BCoV. Positive samples were submitted to sequencing analysis. Bovine RVA and BCoV were circulating in 63% (12/19) and 10.52% (2/19) of the dairy farms, respectively, where 9.5% (46/484) of the calves were positives to RVA and 0.4% (2/484) to BCoV. In beef herds, RVA was detected in 40% (8/20) of the farms and in 6.75% (21/311) of the calves, without positives cases of BCoV. Molecular analysis showed that in dairy farms, G6P[11] and G10P[11] were the prevalent RVA strains, while in beef farms, G10P[11] was the prevalent. The main finding was the detection for the first time of a G15P[11] causing diarrhea in beef calves of Argentina that represents a new alert to be consider for future vaccine updates. Analysis of detected BCoV showed that it is related to the other circulating strains of Argentina.
Assuntos
Doenças dos Bovinos/virologia , Coronavirus Bovino/isolamento & purificação , Diarreia/veterinária , Infecções por Rotavirus/veterinária , Rotavirus/isolamento & purificação , Animais , Animais Recém-Nascidos , Argentina , Bovinos , Coronavirus Bovino/genética , Diarreia/virologia , Ensaio de Imunoadsorção Enzimática/veterinária , Fezes/virologia , Genótipo , Rotavirus/genética , Infecções por Rotavirus/virologiaRESUMO
Bovine coronaviruses (BoCVs) have been found in respiratory tissues in cattle and frequently associated with bovine respiratory disease (BRD); however, pathogenesis studies in calves are limited. To characterize the pathogenesis and pathogenicity of BoCV isolates, we used 5 different BoCV strains to inoculate colostrum-deprived calves, ~ 2-5 wk of age. Later, to determine if dual viral infection would potentiate pathogenicity of BoCV, calves were inoculated with BoCV alone, bovine viral diarrhea virus (BVDV) alone, or a series of dual-infection (BVDV-BoCV) schemes. A negative control group was included in all studies. Clinical signs and body temperature were monitored during the study and samples collected for lymphocyte counts, virus isolation, and serology. During autopsy, gross lesions were recorded and fixed tissues collected for histopathology and immunohistochemistry; fresh tissues were collected for virus isolation. Results suggest increased pathogenicity for isolate BoCV OK 1776. Increased body temperature was found in all virus-inoculated groups. Lung lesions were present in calves in all dual-infection groups; however, lesions were most pronounced in calves inoculated with BVDV followed by BoCV inoculation 6 d later. Lung lesions were consistent with mild-to-moderate interstitial pneumonia, and immunohistochemistry confirmed the presence of BoCV antigen. Our studies demonstrated that BVDV-BoCV dual infection may play an important role in BRD pathogenesis, and timing between infections seems critical to the severity of lesions.
Assuntos
Anticorpos Antivirais/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Coronavirus Bovino/isolamento & purificação , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Doenças Respiratórias/veterinária , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/patologia , Bovinos , Colostro , Diarreia/veterinária , Vírus da Diarreia Viral Bovina/imunologia , Feminino , Gravidez , Doenças Respiratórias/patologia , Doenças Respiratórias/virologiaRESUMO
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.