RESUMO
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.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina , Vírus da Diarreia Viral Bovina Tipo 1 , Vírus da Diarreia Viral Bovina , Surtos de Doenças , Filogenia , Animais , Bovinos , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/prevenção & controle , Surtos de Doenças/veterinária , Feminino , Vírus da Diarreia Viral Bovina Tipo 1/genética , Vírus da Diarreia Viral Bovina Tipo 1/classificação , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Vírus da Diarreia Viral Bovina Tipo 1/imunologia , Brasil/epidemiologia , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/isolamento & purificação , Vírus da Diarreia Viral Bovina/imunologia , Genótipo , Vacinas Virais/imunologia , Ensaio de Imunoadsorção Enzimática , Indústria de Laticínios , Vacinação/veterinária , Anticorpos Antivirais/sangueRESUMO
The envelope glycoprotein E2 of pestiviruses is a major target for neutralizing antibodies. In this study, we analyzed the E2 DA domain of 43 pestiviruses from Southern Brazil. The isolates were identified as Bovine viral diarrhea virus (BVDV) subtypes 1a and 1b or BVDV-2b. Compared to reference strains, the BVDV-1 and -2 isolates had four and two mutations in the DA domain, respectively. All BVDV-2 isolates had a deletion of residues 724 and 725. All mutated amino acids in the BVDV isolates had the same aa substitution, and all were in previously identified antibody binding sites. It is possible that an immunity-mediated selection is acting on the pestiviruses circulating in Southern Brazil.
Assuntos
Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/isolamento & purificação , Proteínas do Envelope Viral/genética , Animais , Antígenos Virais/genética , Sítios de Ligação de Anticorpos/genética , Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Brasil/epidemiologia , Bovinos , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/imunologia , Mutação , Filogenia , RNA Viral/genética , Proteínas do Envelope Viral/imunologiaRESUMO
Detection of bovine viral diarrhea virus (BVDV) in aborted fetus samples is often difficult due to tissue autolysis and inappropriate sampling. Studies assessing different methods for BVDV identification in fetal specimens are scarce. The present study evaluated the agreement between different diagnostic techniques to detect BVDV infections in specimens from a large number of bovine aborted fetuses and neonatal deaths over a period of 22 years. Additionally, genetic, serological, and pathological analyses were conducted in order to characterize BVDV strains of fetal origin. Samples from 95 selected cases from 1997 to 2018 were analyzed by antigen-capture ELISA (AgELISA), nested RT-PCR (RT-nPCR), and real-time RT-PCR (RT-qPCR). In addition, amplification and sequencing of the 5'UTR region were performed for phylogenetic purposes. Virus neutralization tests against the BVDV-1a, BVDV-1b, and BVDV-2b subtypes were conducted on 60 fetal fluids of the selected cases. Furthermore, the frequency and severity of histopathological lesions were evaluated in BVDV-positive cases. This study demonstrated that RT-nPCR and RT-qPCR were more suitable than AgELISA for BVDV detection in fetal specimens. However, the agreement between the two RT-PCR methods was moderate. The BVDV-1b subtype was more frequently detected than the BVDV-1a and BVDV-2b subtypes. Neutralizing antibodies to any of the three subtypes evaluated were present in 94% of the fetal fluids. Microscopically, half of the BVDV-positive cases showed a mild non-suppurative inflammatory response. These results emphasize the need to consider different methods for a diagnostic approach of BVDV associated to reproductive losses.
Assuntos
Feto Abortado/virologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/diagnóstico , Vírus da Diarreia Viral Bovina/classificação , Filogenia , Regiões 5' não Traduzidas , Animais , Anticorpos Neutralizantes/imunologia , Bovinos , Vírus da Diarreia Viral Bovina/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase em Tempo Real , Fatores de TempoRESUMO
Bovine viral diarrhea virus (BVDV) is a major pathogen worldwide, causing significant economic losses to the livestock sector. In Uruguay, BVDV seroprevalence at the farm level is >80%. In this work, 2546 serum, blood or tissue samples collected from animals suspected of being affected by BVD between 2015 and 2017 were analyzed by reverse transcription PCR and sequencing. Analysis of the BVDV genomic regions 5'UTR/Npro, Npro and E2 revealed that BVDV-1a, 1i and 2b circulate in the country, with BVDV-1a being the most prevalent subtype. Population dynamics studies revealed that BVDV-1a has been circulating in our herds since ~1990. This subtype began to spread and evolve, accumulating point mutations at a rate of 3.48 × 10-3 substitutions/site/year, acquiring specific genetic characteristics that gave rise to two local genetic lineages of BVDV-1a. These lineages are divergent from those circulating worldwide, as well as the vaccine strain currently used in Uruguay. The most notable differences between field and vaccine strains were found in the E2 glycoprotein, suggesting that the amino acid substitutions could result in failure of cross-protection/neutralization after vaccination. This is the first study that compares Uruguayan BVDV field and vaccine strains with other BVDV strains from throughout the world. The results obtained in this study will be very useful for developing a suitable immunization program for BVDV in Uruguay by identifying local field strains as candidates for vaccine development.
Assuntos
Vírus da Diarreia Viral Bovina/classificação , Mutação Puntual , Análise de Sequência de RNA/métodos , Substituição de Aminoácidos , Animais , Bovinos , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/imunologia , Evolução Molecular , Filogenia , Estudos Soroepidemiológicos , Uruguai , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/imunologia , Vacinas Virais/imunologiaRESUMO
HoBi-like is an emerging pestivirus of the family Flaviviridae detected in cattle herds and biological products of bovine origin in many parts of the world, causing disease similar to that observed in bovine viral diarrhea virus (BVDV) infections. In this study we reported the detection of HoBi-like pestivirus in an outbreak of respiratory disease in calves from Brazil, seropositive for viruses of the bovine respiratory disease complex (BRDC). Thus, serum samples and nasal swabs were collected from calves up to one year old, presenting or not clinical signs of respiratory disease. Serum samples were submitted to virus neutralization test (VNT) for BVDV-1, BVDV-2, bovine herpesvirus-1 (BoHV-1), bovine respiratory syncytial virus (BRSV) and bovine parainfluenza-3 (BPIV-3). These samples were also tested for the presence of pestiviruses (BVDV-1, BVDV-2 and HoBi-like) and BoHV-1 by RT-PCR and PCR, respectively. Nasal swabs were analyzed by RT-PCR for pestiviruses, BRSV and BPIV-3. VNT results showed high serological prevalence and a wide range of antibodies titers, for all viruses studied, in calves of different age groups. The RT-PCR amplified the 5'UTR and E2 regions of pestiviruses of four calves, from both nasal swabs and serum samples, which sequencing identified the HoBi-like pestivirus. This is the first detection of HoBi-like in nasal secretions of calves in an outbreak of respiratory disease in Brazil, along with the serological detection of other respiratory viruses. We concluded that HoBi-like pestivirus should be considered as part of the BRDC, as a differential diagnosis, to take correct measures of control and prophylaxis.
Assuntos
Doenças dos Bovinos/epidemiologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Surtos de Doenças/veterinária , Infecções por Pestivirus/veterinária , Doenças Respiratórias/veterinária , Animais , Complexo Respiratório Bovino/epidemiologia , Complexo Respiratório Bovino/virologia , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/virologia , Vírus da Diarreia Viral Bovina/classificação , Feminino , Infecções por Pestivirus/epidemiologia , Infecções por Pestivirus/virologia , Prevalência , Doenças Respiratórias/epidemiologia , Doenças Respiratórias/virologia , Estudos SoroepidemiológicosRESUMO
BACKGROUND: Bovine Viral Diarrhea Virus (BVDV) is associated with gastrointestinal, respiratory and reproductive diseases of livestock across the world that causes continuous economic losses in the cattle industry. This virus can establish a persistent infection (PI) in calves after the fetal infection, making BVDV positive catle carriers and primary reservoirs which will constantly transmit the virus to healthy and new-born animals. For this reason, the detection of the PI animals in herds is the first line of prevention of the viral infection. RESULTS: In this study, PI animals were detected in five different regions of Colombia through RT-PCR techniques and confirmed by sequencing. BVDV genotypes were determined using one fragment of the 5'UTR. It was found a 7% BVDV prevalence in animals and 22% in farms; and genotype 1 was identified as a single genotype for all of the samples. All samples were BVDV 1a. CONCLUSION: This is the first report in Colombia with higher prevalence rates compared with other places in the world, turned out to be of great importance for the ranchers, the vaccine producers and animal health control parties.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/prevenção & controle , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Bovinos , Colômbia/epidemiologia , Vírus da Diarreia Viral Bovina/classificação , Feminino , Masculino , Filogenia , Prevalência , Reação em Cadeia da Polimerase em Tempo Real/veterináriaRESUMO
Dermatitis might occur in mucosal disease (MD) caused by bovine viral diarrhea virus (BVDV). This study describes the pathological and virological features of skin lesions associated with BVDV infection in four persistently infected (PI) cattle. Skin samples were reprocessed for histopathology and IHC. BVDV isolates were obtained and were genetically characterized. In addition to upper alimentary system ulcerative lesions, all cattle (one outbreak and three individual cases) presented focal crusty and ulcerative lesions affecting the mucocutaneous and skin-horn junctions, interdigital clefts, pastern, and areas surrounding the dewclaws and diffuse thickened skin within 7-20 days of infection. Microscopic analysis revealed parakeratotic hyperkeratosis and single-cell keratinocyte death, accompanied by ballooning degeneration and spongiosis in the epidermis, as well as intraepithelial and subcorneal pustules. IHC showed BVDV antigen in the cytoplasm of keratinocytes undergoing individual cell death. Phylogenetic analysis revealed that the isolates from cattle #1, #2, and #4 belonged to BVDV-1a, whereas that from cattle #3 belonged to BVDV-1d. Cytopathic BVDV was isolated from cattle #2 and #3 (MD), and non-cytopathic BVDV was isolated from cattle #1 and #4. Thus, BVDV infection might cause acute disease, characterized by skin and upper alimentary system ulcerative lesions, in both MD and PI cattle.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Dermatopatias/veterinária , Animais , Anticorpos Antivirais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/patologia , Bovinos , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/fisiologia , Filogenia , Pele/patologia , Pele/virologia , Dermatopatias/patologia , Dermatopatias/virologiaRESUMO
Hobi-like viruses (HobiPeV) comprise a novel, recently classified species of bovine pestiviruses, originally identified in commercial fetal bovine serum of Brazilian origin and, subsequently, isolated from diseased animals in several countries. Although frequently isolated from clinical cases, most HobiPeV isolates failed to reproduce overt disease in cattle upon experimental inoculation. Herein, we describe the outcome of experimental infection of four to six months-old seronegative calves with two Brazilian HobiPeV isolates. Calves inoculated intranasally with isolate SV478/07 developed viremia between days 2 and 9 post-inoculation (pi) and shed virus in nasal secretions up to day 11pi. These animals presented hyperthermia (day 7 to 10-11 pi) and lymphopenia from days 4 to 8pi. Clinically, all four calves developed varied degrees of apathy, anorexia, mild to moderate respiratory signs (nasal secretion, hyperemia), ocular discharge and pasty diarrhea in the days following virus inoculation. In contrast, calves inoculated with isolate SV757/15 presented only hyperthermia (days 3 to 10-11 pi) and lymphopenia (days 4-8 pi), without other apparent clinical signs. In these animals, viremia was detected up to day 9 pi and virus shedding in nasal secretions lasted up to day 12-14 pi. Both groups seroconverted to the inoculated viruses, developing virus neutralizing (VN) titers from 320 to 5120â¯at day 28pi. These results extend previous findings that experimental infections of calves with HobiPeV are predominantly mild, yet they also indicate that field isolates may differ in their ability to cause disease in susceptible animals.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Doenças dos Bovinos/virologia , Bovinos/virologia , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/patogenicidade , Febre/virologia , Linfopenia/virologia , Infecções por Pestivirus/virologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Temperatura Corporal , Doença das Mucosas por Vírus da Diarreia Viral Bovina/imunologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/fisiopatologia , Brasil , Vírus da Diarreia Viral Bovina/isolamento & purificação , Modelos Animais de Doenças , Masculino , Infecções por Pestivirus/imunologia , Infecções por Pestivirus/veterinária , Fatores de Tempo , Carga Viral , Viremia/virologia , Eliminação de Partículas ViraisRESUMO
The ruminant pestiviral species BVDV-1, BVDV-2 and BDV, along with the putative species HoBi-like, may cause substantial economic losses in cattle, sheep and goats. Brazil's large size, variable biomes and wide range of ruminant animal production within different geographic regions suggest that the presence and prevalence of ruminant pestivirus may differ by regions within Brazil. This study investigated the genetic diversity of ruminant pestiviruses and determined the frequency of active infections within two states of the Northeast Region of Brazil, Maranhão and Rio Grande do Norte. Serum samples from 16,621 cattle and 2,672 small ruminants from 569 different herds residing in this region were tested by RT-PCR followed by DNA sequencing. Seventeen positive cattle were detected (0.1%) from fifteen different herds (2.64%). All isolates were classified as HoBi-like pestiviruses based on phylogenetic analysis. All small ruminant samples tested negative. The findings presented herein suggest that the Northeast Region of Brazil has a uniquely high prevalence of HoBi-like viruses. The increasing reports of HoBi-like viruses detected in cattle in the field suggest that natural infection with these viruses may be more widespread than previously thought. The identification of HoBi-like viruses as the most prevalent type of ruminant pestivirus circulating in the Northeast Region of Brazil indicates the need for both continued monitoring and determination of the extent of economic losses associated with HoBi-like virus infections. In addition, it must be taken into account in the choice of diagnostic tests and in vaccine formulations.
Assuntos
Doenças dos Bovinos/virologia , Vírus da Diarreia Viral Bovina/genética , Variação Genética , Infecções por Pestivirus/veterinária , Animais , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Vírus da Diarreia Viral Bovina Tipo 1/classificação , Vírus da Diarreia Viral Bovina Tipo 1/genética , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Vírus da Diarreia Viral Bovina Tipo 2/classificação , Vírus da Diarreia Viral Bovina Tipo 2/genética , Vírus da Diarreia Viral Bovina Tipo 2/isolamento & purificação , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/isolamento & purificação , Infecções por Pestivirus/epidemiologia , Infecções por Pestivirus/virologia , Filogenia , Prevalência , Ruminantes , Análise de Sequência de DNA/veterináriaRESUMO
The present study performed a genetic identification of pestiviruses contaminating batches of fetal bovine serum (FBS) produced in Brazil from 2006 to 2014. Seventy-three FBS lots were screened by a RT-PCR targeting the 5'untranslated region (UTR) of the pestivirus genome. Thirty-nine lots (53.4%) were positive for pestivirus RNA and one contained infectious virus. Nucleotide sequencing and phylogenetic analysis of the 5'UTR revealed 34 lots (46.6%) containing RNA of bovine viral diarrhea virus type 1 (BVDV-1), being 23 BVDV-1a (5' UTR identity 90.8-98.7%), eight BVDV-1b (93.9-96.7%) and three BVDV-1d (96.2- 97.6%). Six lots (8.2%) contained BVDV-2 (90.3-100% UTR identity) being two BVDV-2a; three BVDV-2b and one undetermined. Four FBS batches (5.5%) were found contaminated with HoBi-like virus (98.3 to 100%). Five batches (6.8%) contained more than one pestivirus. The high frequency of contamination of FBS with pestivirus RNA reinforce the need for systematic and updated guidelines for monitoring this product to reduce the risk of contamination of biologicals and introduction of contaminating agents into free areas.(AU)
No presente estudo foi realizada a identificação genética de pestivírus contaminantes de lotes de soro fetal bovino (SFB) produzidos no Brasil de 2006 a 2014. Setenta e três lotes de SFB foram testados por RT-PCR para a região 5' não traduzida do genoma dos pestivírus. Trinta e nove lotes (53,4%) foram positivos para RNA de pestivírus e um continha vírus infeccioso. O sequenciamento de nucleotídeos e análise filogenética da região 5'UTR revelou que 34 lotes (46,6%) continham RNA do vírus da diarreia viral bovina tipo 1 (BVDV-1), sendo 23 BVDV-1a (identidade na 5' UTR de 90,8-98,7%), oito BVDV-1b (93,9 a 96,7%) e três BVDV-1d (96,2%-97,6%). Seis lotes (8,2%) continham BVDV-2 (90,3 a 100% de identidade), sendo dois BVDV-2a, três BVDV-2b e um de subgenótipo indeterminado. Quatro lotes de SFB (5,5%) estavam contaminados com o vírus HoBi-like (98,3 a 100%). Cinco lotes (6,8%) continham mais do que um pestivírus. A alta frequência de contaminação de SFB com RNA de pestivírus reforça a necessidade para diretrizes sistemáticas atualizadas para a monitoração deste produto com a finalidade de reduzir a contaminação de produtos biológicos e a introdução de agentes contaminantes em áreas livres.(AU)
Assuntos
Animais , Bovinos , Bovinos/virologia , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/genéticaRESUMO
The present study performed a genetic identification of pestiviruses contaminating batches of fetal bovine serum (FBS) produced in Brazil from 2006 to 2014. Seventy-three FBS lots were screened by a RT-PCR targeting the 5'untranslated region (UTR) of the pestivirus genome. Thirty-nine lots (53.4%) were positive for pestivirus RNA and one contained infectious virus. Nucleotide sequencing and phylogenetic analysis of the 5'UTR revealed 34 lots (46.6%) containing RNA of bovine viral diarrhea virus type 1 (BVDV-1), being 23 BVDV-1a (5' UTR identity 90.8-98.7%), eight BVDV-1b (93.9-96.7%) and three BVDV-1d (96.2- 97.6%). Six lots (8.2%) contained BVDV-2 (90.3-100% UTR identity) being two BVDV-2a; three BVDV-2b and one undetermined. Four FBS batches (5.5%) were found contaminated with HoBi-like virus (98.3 to 100%). Five batches (6.8%) contained more than one pestivirus. The high frequency of contamination of FBS with pestivirus RNA reinforce the need for systematic and updated guidelines for monitoring this product to reduce the risk of contamination of biologicals and introduction of contaminating agents into free areas.(AU)
No presente estudo foi realizada a identificação genética de pestivírus contaminantes de lotes de soro fetal bovino (SFB) produzidos no Brasil de 2006 a 2014. Setenta e três lotes de SFB foram testados por RT-PCR para a região 5' não traduzida do genoma dos pestivírus. Trinta e nove lotes (53,4%) foram positivos para RNA de pestivírus e um continha vírus infeccioso. O sequenciamento de nucleotídeos e análise filogenética da região 5'UTR revelou que 34 lotes (46,6%) continham RNA do vírus da diarreia viral bovina tipo 1 (BVDV-1), sendo 23 BVDV-1a (identidade na 5' UTR de 90,8-98,7%), oito BVDV-1b (93,9 a 96,7%) e três BVDV-1d (96,2%-97,6%). Seis lotes (8,2%) continham BVDV-2 (90,3 a 100% de identidade), sendo dois BVDV-2a, três BVDV-2b e um de subgenótipo indeterminado. Quatro lotes de SFB (5,5%) estavam contaminados com o vírus HoBi-like (98,3 a 100%). Cinco lotes (6,8%) continham mais do que um pestivírus. A alta frequência de contaminação de SFB com RNA de pestivírus reforça a necessidade para diretrizes sistemáticas atualizadas para a monitoração deste produto com a finalidade de reduzir a contaminação de produtos biológicos e a introdução de agentes contaminantes em áreas livres.(AU)
Assuntos
Animais , Bovinos , Bovinos/virologia , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/genéticaRESUMO
Bovine viral diarrhea (BVD) affects bovine production and reproduction causing significant economic losses all over the world. Two viral species has been recognized: BVDV-1 and BVDV-2, both distributed worldwide. Recently, novel specie of BVDV named HoBi-like pestivirus was discovered. The presence of BVDV was confirmed in 1996 in Uruguay, however, does not exist until today a schedule of compulsory vaccination along the country. Serological studies with samples from all Uruguayan herds were performed during 2000 and 2001 demonstrating that all of them were seropositive to BVDV with a mean prevalence of 69%. In addition, there have been no new studies done since those previously described and it is important to mention that the genetic diversity of BVD has never been described in Uruguay. Nowadays, there is strongly suspect that BVDV is one of the most important causes of reproductive failures in our herds. The aim of this study was to describe for the first time in Uruguay the genetic diversity of BVDV with samples collected from different regions along the country. Serological status of 390 non-vaccinated animals against BVDV with reproductive problems from farms of Rivera, Tacuarembó and Florida departments of Uruguay were studied. All herds were seropositive to BVDV and high proportion of animals were positive (298/390), while 4.1% (16/390) of the animals were positive to Antigen Capture ELISA test and Real Time PCR. Phylogenetic analysis performed with concatenated sequences from the 5'UTR and Npro genomic regions revealed that BVDV-1 and BVDV-2 are infecting our herds, being BVDV-1 the most frequently found. The major subtype was BVDV-1a, followed by BVDV-1i and BVDV-2b. This is the first study that describes the genetic diversity of BVDV in Uruguay and it will contribute to the elaboration of sanitization programs.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/genética , Variação Genética , Regiões 5' não Traduzidas , Animais , Bovinos , Análise por Conglomerados , Vírus da Diarreia Viral Bovina/isolamento & purificação , Genótipo , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Homologia de Sequência , Uruguai , Proteínas Virais/genéticaRESUMO
Bovine viral diarrhoea virus (BVDV) causes one of the most important diseases of cattle in terms of economic costs and welfare. The aims were to estimate herd prevalence and to investigate the factors associated with antibodies in bulk tank milk (BTM) in dairy herds through a matched case-control study. To estimate herd prevalence, BTM samples were randomly selected (n = 314) from a population (N = 1604). The true prevalence of BVDV was 24.3% (CI 95% = 20.1-29.3%). For the case-control study, BVDV antibody-positive herds (high antibody titres) were classified as cases (n = 21) and matched (n = 63) by milk production with herds presenting low antibody titres (ratio of 1 : 3). Three multivariable models were built: 1) full model, holding all 21 variables, and two models divided according to empirical knowledge and similarity among variables; 2) animal factor model; and 3) biosecurity model. The full model (model 1) identified: age as a culling criteria (OR = 0.10; CI 95% = 0.02-0.39; P < 0.01); farms that provided milk to other industries previously (OR = 4.13; CI 95% = 1.17-14.49; P = 0.02); and isolation paddocks for ill animals (OR = 0.14; CI 95% = 0.01-0.26; P = 0.02). The biosecurity model revealed a significant association with the use of natural mating (OR = 9.03; CI 95% = 2.14-38.03; P < 0.01); isolation paddocks for ill animals (OR = 0.06; CI 95% = 0.05-0.83; P = 0.03); years providing milk for the same industry (OR = 0.94; CI 95% = 0.91-0.97; P = 0.02); and direct contact over fences among cattle of neighbouring farms (OR = 5.78; CI 95% = 1.41-23.67; P = 0.04). We recommend the application of grouping predictors as a good choice for model building because it could lead to a better understanding of disease-exposure associations.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Animais , Anticorpos Antivirais/análise , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Brasil/epidemiologia , Estudos de Casos e Controles , Bovinos , Estudos Transversais , Vírus da Diarreia Viral Bovina/classificação , Leite/virologia , Análise Multivariada , Prevalência , Fatores de RiscoRESUMO
Bovine viral diarrhoea virus (BVDV) is an important pathogen of cattle that occurs worldwide with substantial economic impact on beef and dairy industries. The aim of this study was to describe the diversity of BVDV subgenotypes in persistently infected (PI) animals identified in a highly productive, regularly vaccinated, dairy cattle herd presenting with reproductive failure. Serum samples were collected from all animals within the herd (n = 692) and used to detect the presence of BVDV RNA. Using reverse transcription polymerase chain reaction assay, 29 cows were identified as transiently infected, three animals (two cows and one calf) as persistently infected, and one calf as putative BVDV PI animal. The sequences of 5'UTR and/or N(pro) gene of BVDV used in phylogenetic analyses revealed that the three PI animals were infected by three different BVDV subgenotypes (BVDV-1a, BVDV-1b, and BVDV-1d). These results demonstrated that in an open dairy cattle herd, regular vaccination against BVDV by itself is not able to prevent viral circulation in the herd. Furthermore, depending on the frequency of the acquisition of heifers and/or cows for replacement, several BVDV subgenotypes may co-exist simultaneously in the same herd.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/prevenção & controle , Vírus da Diarreia Viral Bovina/genética , Vacinas Virais , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Brasil , Bovinos , Indústria de Laticínios , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/imunologia , Feminino , Variação Genética , Genótipo , Masculino , FilogeniaRESUMO
A infecção natural pelo vírus da diarréia viral bovina (BVDV) foi monitorada em três rebanhos bovinos por meio de amostras de soro sangüíneo, obtidas em várias colheitas realizadas em cada rebanho, que foram submetidas ao teste de virusneutralização (VN) para o BVDV-1 e para o BVDV-2. As amostras não reagentes a pelo menos um dos genótipos e aquelas oriundas de bovinos com menos de seis meses de idade, reagentes ou não, foram analisadas pela reação em cadeia da polimerase precedida pela transcrição reversa (RT-PCR) para a pesquisa do BVDV. Em dois rebanhos o vírus não foi detectado em nenhuma amostra e a quantidade de animais reagentes ao vírus no teste de VN, principalmente nos animais jovens, diminuiu à medida que as colheitas foram realizadas. No entanto, no terceiro rebanho, a infecção permaneceu durante o período monitorado, pois o BVDV foi detectado em dois animais persistentemente infectados (PI) e também em um animal transitoriamente infectado (TI). O sistema de criação, bem como o intenso trânsito de animais, foram favoráveis à permanência da infecção nesse último rebanho. A dinâmica da infecção pelo BVDV foi variável nos rebanhos analisados, destacando a provável eliminação espontânea do BVDV no rebanho 1 e os fatores de risco relacionados à transmissão do BVDV, como a freqüente aquisição de animais de diversas procedências pelo rebanho 4, assim como a provável hipótese da infecção do rebanho 17 ter originado a partir do rebanho vizinho.(AU)
Natural infection by bovine viral diarrhea virus (BVDV) was monitored in blood serum samples of three cattle herds. The samples were drawn in several harvests from each herd and submitted to virus neutralization test (VN) against BVDV-1 and BVDV-2. The non reactive samples to at least one of the genotypes and also those collected from calves younger than six months, were analyzed by reverse transcription polymerase chain reaction (RT-PCR). In two herds, BVDV was not detected in any blood sample and the number of reactive samples to VN test, especially young animals, decreased as the blood sample harvests were conducted. However, in the third herd, the infection remained during the monitored period, because BVDV was detected in two persistently infected animals (PI) and also in one transiently infected animal (TI). The cattle breeding system and intense movement of animals were favorable to the permanence of infection in this last herd. The dynamics of BVDV infection changed in the analyzed herds, highlighting the probable self clearance of BVDV in herd 1 and the risk factors related to transmission of BVDV, such as the frequent purchase of animals from different origins for herd 4, as well as the probable hypothesis that infection of herd 17 may have originated from the neighboring herd.(AU)
Assuntos
Animais , Bovinos , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/crescimento & desenvolvimentoRESUMO
A infecção natural pelo vírus da diarréia viral bovina (BVDV) foi monitorada em três rebanhos bovinos por meio de amostras de soro sangüíneo, obtidas em várias colheitas realizadas em cada rebanho, que foram submetidas ao teste de virusneutralização (VN) para o BVDV-1 e para o BVDV-2. As amostras não reagentes a pelo menos um dos genótipos e aquelas oriundas de bovinos com menos de seis meses de idade, reagentes ou não, foram analisadas pela reação em cadeia da polimerase precedida pela transcrição reversa (RT-PCR) para a pesquisa do BVDV. Em dois rebanhos o vírus não foi detectado em nenhuma amostra e a quantidade de animais reagentes ao vírus no teste de VN, principalmente nos animais jovens, diminuiu à medida que as colheitas foram realizadas. No entanto, no terceiro rebanho, a infecção permaneceu durante o período monitorado, pois o BVDV foi detectado em dois animais persistentemente infectados (PI) e também em um animal transitoriamente infectado (TI). O sistema de criação, bem como o intenso trânsito de animais, foram favoráveis à permanência da infecção nesse último rebanho. A dinâmica da infecção pelo BVDV foi variável nos rebanhos analisados, destacando a provável eliminação espontânea do BVDV no rebanho 1 e os fatores de risco relacionados à transmissão do BVDV, como a freqüente aquisição de animais de diversas procedências pelo rebanho 4, assim como a provável hipótese da infecção do rebanho 17 ter originado a partir do rebanho vizinho.
Natural infection by bovine viral diarrhea virus (BVDV) was monitored in blood serum samples of three cattle herds. The samples were drawn in several harvests from each herd and submitted to virus neutralization test (VN) against BVDV-1 and BVDV-2. The non reactive samples to at least one of the genotypes and also those collected from calves younger than six months, were analyzed by reverse transcription polymerase chain reaction (RT-PCR). In two herds, BVDV was not detected in any blood sample and the number of reactive samples to VN test, especially young animals, decreased as the blood sample harvests were conducted. However, in the third herd, the infection remained during the monitored period, because BVDV was detected in two persistently infected animals (PI) and also in one transiently infected animal (TI). The cattle breeding system and intense movement of animals were favorable to the permanence of infection in this last herd. The dynamics of BVDV infection changed in the analyzed herds, highlighting the probable self clearance of BVDV in herd 1 and the risk factors related to transmission of BVDV, such as the frequent purchase of animals from different origins for herd 4, as well as the probable hypothesis that infection of herd 17 may have originated from the neighboring herd.
Assuntos
Animais , Bovinos , Vírus da Diarreia Viral Bovina/classificação , Vírus da Diarreia Viral Bovina/crescimento & desenvolvimentoRESUMO
Bovine pestiviruses (Bovine Viral Diarrea Virus 1 (BVDV 1) and Bovine Viral Diarrea Virus 2 (BVDV 2)) belong to the genus Pestivirus (Flaviviridae), which is composed of positive stranded RNA viruses causing significant economic losses world-wide. We used phylogenetic and bootstrap analyses to systematically scan alignments of previously sequenced genomes in order to explore further the evolutionary mechanisms responsible for variation in the virus. Previously published data suggested that homologous crossover might be one of the mechanisms responsible for the genomic rearrangements observed in cytopathic (cp) strains of bovine pestiviruses. Nevertheless, homologous recombination involves not just homologous crossovers, but also replacement of a homologous region of the acceptor RNA. Furthermore, cytopathic strains represent dead paths in evolution, since they are isolated exclusively from the fatal cases of mucosal disease. Herein, we report evidence of homologous inter-genotype recombination in the genome of a non-cytopathic (ncp) strain of Bovine Viral Diarrea Virus 1, the type species of the genus Pestivirus. We also show that intra-genotype homologous recombination might be a common phenomenon in both species of Pestivirus. This evidence demonstrates that homologous recombination contribute to the diversification of bovine pestiviruses in nature. Implications for virus evolution, taxonomy and phylogenetics are discussed.
Assuntos
Vírus da Diarreia Viral Bovina/genética , RNA Viral/genética , Recombinação Genética , Algoritmos , Sequência de Aminoácidos , Animais , Bovinos , Vírus da Diarreia Viral Bovina/classificação , Evolução Molecular , Variação Genética , Genoma Viral , Filogenia , Homologia de Sequência de AminoácidosRESUMO
The objective of the present work is the description outbreaks caused by bovine viral diarrhea virus (BVDV) in commercial beef cattle ranches in Argentina. Genetic affiliation and their association with the clinical manifestation were carried out with five BVDV isolates from an outbreak of mucosal disease (MD) (Outbreak #1), acute enteritis (Outbreaks #2 and #3) and generalized dermatitis (Outbreaks #4 and #5). Upon genetic analysis CP BVDV isolate of Outbreak #1 clustered to closely to BVDV Oregon (Genotype 1). BVDV isolates from the outbreaks of generalized dermatitis (Outbreaks #4 and #5) were located close to BVDV Osloss within Genotype 1. The identification by immunohistochemistry of BVDV in exudative dermatitis indicates the epithelial cell tropism of the virus. Phylogenic characterization of BVDV from Outbreaks #2 and #3 locate them as BVDV-2. 5'UTR sequence of these viruses revealed a homology of 88 and 90% to BVDV-890 (Genotype 2) and a 77 and 75% to BVDV-SD1 (Genotype 1), respectively. The association of BVDV-2 with severe disease indicates the presence of highly virulent strains. Data from natural outbreaks where BVDV-1 and BVDV-2 were isolated revealed that pathology overlaps and not clearly allows the differentiation between genotypes based on gross or microscopic lesions. Thus, for a definitive diagnosis, further virology and molecular studies are necessary. Additionally, the results of this work focused on the origin and consequences of genetic variations of BVDV with regard to pathogenesis and suggest the association between genotype and a defined clinical syndrome.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina/genética , Surtos de Doenças/veterinária , Animais , Argentina/epidemiologia , Sequência de Bases , Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/patologia , Bovinos , DNA Viral/análise , DNA Viral/química , Vírus da Diarreia Viral Bovina/classificação , Feminino , Genótipo , Imuno-Histoquímica/veterinária , Masculino , Dados de Sequência Molecular , Testes de Neutralização/veterinária , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Homologia de Sequência do Ácido NucleicoRESUMO
Phylogenetic analysis divides bovine viral diarrhea viruses (BVDV) into two different genotypes (BVDV1 and BVDV2). BVDV1 strains have been further subdivided into two to 11 subgenotypes. Phylogenetic analysis of BVDV2 isolates, however, has not been able to identify discrete subgenotypes. In this study, we identified six South American BVDV2 strains and one North American BVDV2 strain that cluster to a separate genetic group within BVDV2, thus representing a distinct subgenotype. The 5' untranslated region (UTR) sequence homology between these six strains and other BVDV2 from North America, Europe and Asia (81.7%) is lower than the homology used to segregate BVDV1 into BVDV1a and BVDV1b (83.6%). Most nucleotide differences observed between the two subgroups of BVDV2 were concentrated in two regions, which also harbor most of the differences seen between BVDV1a and BVDV1b. To determine if this segregation was real, an additional analysis was performed comparing NS2/3 sequences. Analysis of a conserved sequence located between nucleotides 6670 and 7186 of the NS2/3 coding region also segregated these isolates to a separate group. The sequence homology between the two subgroups (86.3%) was higher than the homology in the 5'UTR (81.7%), with mean sequence homologies of 91 and 87.2% within the proposed subgroups. In contrast to the 5'UTR, alignment of the NS2/3 sequences revealed nucleotide differences distributed across the region. These results demonstrate that BVDV2 isolates cluster to two genetically distinct subgroups within BVDV2. The differences in both the 5'UTR and NS2/3 are consistent and justify this segregation. We suggest that BVDV2 may thereafter be subgenotyped into BVDV2a and BVDV2b. The existence of subgroups within the BVDV2 genotype with genetic heterogeneity similar to that seen among BVDV1 subgroups argues against BVDV2 isolates arising from BVDV1 in a recent evolutionary event. Unless the evolutionary clocks for BVDV1 and BVDV2 isolates tick along at different rates, these results indicate that BVDV2 have existed as long as BVDV1.
Assuntos
Vírus da Diarreia Viral Bovina/classificação , Regiões 5' não Traduzidas , Animais , Sequência de Bases , Brasil , Bovinos , DNA Viral , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/isolamento & purificação , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNARESUMO
Genetic typing of 29 Bovine Viral Diarrhea Virus (BVDV) isolates from Argentina was carried out by sequencing 245 nucleotides of the RT-PCR products of the 5'-UTR region. Sequence analysis shows that these Argentinean BVDV include types 1 and 2. The majority (26/29) of the isolates are type 1, which comprises subtypes 1a and 1b, together with an additional subgroup within subtype 1a. This subgroup is close to the South African subgroup Ic of 1a viruses, and to the deer pestivirus strain "Deer". The three type 2 BVDV were isolated from fetal tissues or serum during the 7-8 years before a clinical outbreak in Argentina had been reported. Only inactivated vaccines are used in bovines of the country, thus the analysed viruses are authentic field strains. The long term circulation of type 2 BVDV (situation similar to that of North America before the epidemic of 1993), and the existence of viral populations which differ from the reference strains commonly used in vaccine elaboration should be considered by manufacturers of diagnostic reagents and vaccines.