RESUMO
The overwhelming majority of SARS-CoV-2 epidemiological studies cover a narrow time period, making general knowledge about the COVID-19 pandemic difficult. To assess COVID-19-related host aspects in the overall pandemic, we analyzed COVID-19 cases during the first two years of SARS-CoV-2 circulation in southern Brazil. Herein, 390 patients admitted in 2020-2022 to a Brazilian public referral hospital were allocated into two groups according to the COVID-19 outcome: hospital discharge (n=237) or death (n=153). In the univariate analysis, several variables, including sociodemographic, clinical and laboratory aspects (primary data), were significantly different between the analyzed groups. In multivariate logistic regression, eight of these factors remained associated with the COVID-19 outcome. In particular, we report oxygen supplementation and the need for hemodialysis as predictors of hospital discharge and death from COVID-19, respectively. To the best of our knowledge, none of these findings have been previously reported in the Brazilian or world population. In conclusion, our results contribute to current knowledge by demonstrating that factors described at different times may remain associated with COVID-19 over the pandemic and by identifying novel predictors of COVID-19 outcome.
Assuntos
COVID-19 , SARS-CoV-2 , Humanos , COVID-19/epidemiologia , Brasil/epidemiologia , Pandemias , HospitalizaçãoRESUMO
This study aimed to evaluate the demographic and healthcare situation of dogs and cats owned by families assisted by the Family Health Strategy (FHS), from Santa Maria/RS, Brazil. This research was a cross-sectional and population-based study developed by applying a questionnaire to residents in the 16 FHS areas of the city. This was the first study addressing pet animal conditions in the FHS area. A total of 414 households were studied, and 88.5% of them had pets (dogs and/or cats), with an average of 2.2 dogs and 0.8 cats per household. Only 18.4% (228/1.241) of the animals were sterilized (dogs, 15.1% [135/891]; cats, 26.7% [93/348]). When considering the number of dogs, households with one resident had fewer dogs than households with two or more residents (p=0.006). The level of education and family income were not associated with the number of animals (p>0.001). However, higher levels of education and family income were associated with the sterilization of dogs, veterinary monitoring, vaccination, and treatment of ectoparasites in dogs and cats (p<0.0001). Additionally, the higher family income was associated with a higher frequency of endoparasite treatment (p<0.05). The study shows a high average number of pets per household in FSH areas compared to the average household population as well as a lack of veterinary care, making it essential to promote responsible custody.
Este estudo teve como objetivo avaliar a situação demográfica e de saúde de cães e gatos pertencentes a famílias assistidas pela Estratégia de Saúde da Família (ESF), de Santa Maria/RS, Brasil. Esta pesquisa foi um estudo transversal baseado na população adscrita, desenvolvido por meio da aplicação de questionário aos residentes das 16 áreas de ESF da cidade. Este foi o primeiro estudo que abordou as condições dos animais de estimação na área de ESF. Foram estudados 414 domicílios, e 88,5% deles tinham animais de estimação (cães e/ou gatos), com uma média de 2,2 cães e 0,8 gatos por domicilio. Apenas 18,4% (228/1,241) dos animais foram esterilizados (cães, 15,1% [135/891]; gatos, 26,7% [93/348]). Ao considerar o número de cães, os lares com um residente tinham menos cães que os lares com dois ou mais residentes (p=0,006). O nível de educação e a renda familiar não estavam associados ao número de animais (p>0,001). Entretanto, níveis mais altos de educação e renda familiar foram associados com a esterilização de cães, monitoramento veterinário, vacinação e tratamento de ectoparasitas em cães e gatos (p<0,0001). Adicionalmente, a maior renda familiar foi associada a uma maior freqüência de tratamento endoparasitário (p<0,05). O estudo mostra um alto número médio de animais de estimação por domicílio nas áreas de ESF em comparação com a população domiciliar média, bem como uma falta de cuidados veterinários, tornando essencial promover a custódia responsável.
Assuntos
Humanos , Animais , Gatos , Cães , Vínculo Humano-Animal , Enquete Socioeconômica , Criação de Animais Domésticos/métodos , Criação de Animais Domésticos/estatística & dados numéricos , Serviço Social/estatística & dados numéricos , BrasilRESUMO
In Latin America, hematophagous bats are the main reservoirs of rabies virus (RABV) to livestock, to other mammals and, occasionally, to human. Nonetheless, reports of exposure of human and pets to RABV upon aggression by non-hematophagous bats are increasing, possibly facilitated by the synanthropic habits of these bats. We, herein, report the detection and genetic identification of a RABV recovered from an insectivorous bat found sick in a student housing building at the Federal University of Santa Maria, Southern Brazil. Taxonomic characterization identified the captured bat as a member of the genus Nyctinomops, family Molossidae, the group of insectivorous bats. Brain fragments of the bat were positive for RABV antigens by fluorescent antibody test (FAT) and for sequences of the nucleoprotein (N) gene by RT-PCR. The N amplicon was submitted to nucleotide sequencing and analysis, showing that the consensus sequences (SV 33/19) had high identity with RABV sequences of insectivorous bats deposited in GenBank. At phylogenetic tree, the N gene sequences of SV 33/19 clustered with RABV recovered from Nyctinomops laticaudatus, Molossus molossus, and Tadarida lauticaudata bats, and a part of RABV variant 3, 4, and 6, that correspond to Desmodus rotundus, Tadarida brasiliensis, and Lasiurus cinereus, respectively. Although no direct human or domestic animal exposure has been reported, this case strengthens the need for a continuous rabies vaccination in pets in the surrounding areas, since non-hematophagous bats may serve as source of infection for these animals. These findings also call attention for continuous monitoring of populations of synanthropic bats to avoid/prevent human exposure.
Assuntos
Quirópteros , Vírus da Raiva , Raiva , Animais , Brasil , Quirópteros/virologia , Filogenia , Raiva/veterinária , Vírus da Raiva/genéticaRESUMO
A emergência de infecções por coronavírus em humanos já foi documentada diversas vezes nas últimas décadas. Estudos filogenéticos indicaram que os coronavírus humanos (human coronavirus - HCoV) de transmissão comunitária e de baixa patogenicidade, como o HCoV-229E e HCoV-OC43, nos anos de 1966/67, e do HCoV-HKU1 e HCoV-NL63, no ano de 2004, se originaram em morcegos e/ou roedores. Os coronavírus causadores das síndromes respiratórias agudas severas (Severe acute respiratory syndrome SARS-CoV) originaram-se de morcegos e, possivelmente, infectaram humanos a partir de hospedeiros intermediários. O gato de algália (Paguma larvata) é o hospedeiro intermediário do SARS-CoV que emergiu no ano de 2002, e o camelo dromedário (Camelus dromedarius) é o hospedeiro intermediário do Middle East respiratory syndrome coronavírus (MERS-CoV), que surgiu em 2012. O hospedeiro intermediário do SARS-CoV-2 ainda é incerto, mas acredita-se que seja o pangolin (Manis javalica). O contato direto ou indireto de humanos com os hospedeiros intermediários, ou com suas secreções e excreções, ou ainda a manipulação e/ou consumo de carnes infectadas provavelmente permitiu o salto deste vírus para a espécie humana.(AU)
Assuntos
Animais , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/patogenicidade , Síndrome Respiratória Aguda Grave/epidemiologia , Síndrome Respiratória Aguda Grave/transmissão , Síndrome Respiratória Aguda Grave/veterinária , Animais de Estimação/virologiaRESUMO
A emergência de infecções por coronavírus em humanos já foi documentada diversas vezes nas últimas décadas. Estudos filogenéticos indicaram que os coronavírus humanos (human coronavirus - HCoV) de transmissão comunitária e de baixa patogenicidade, como o HCoV-229E e HCoV-OC43, nos anos de 1966/67, e do HCoV-HKU1 e HCoV-NL63, no ano de 2004, se originaram em morcegos e/ou roedores. Os coronavírus causadores das síndromes respiratórias agudas severas (Severe acute respiratory syndrome SARS-CoV) originaram-se de morcegos e, possivelmente, infectaram humanos a partir de hospedeiros intermediários. O gato de algália (Paguma larvata) é o hospedeiro intermediário do SARS-CoV que emergiu no ano de 2002, e o camelo dromedário (Camelus dromedarius) é o hospedeiro intermediário do Middle East respiratory syndrome coronavírus (MERS-CoV), que surgiu em 2012. O hospedeiro intermediário do SARS-CoV-2 ainda é incerto, mas acredita-se que seja o pangolin (Manis javalica). O contato direto ou indireto de humanos com os hospedeiros intermediários, ou com suas secreções e excreções, ou ainda a manipulação e/ou consumo de carnes infectadas provavelmente permitiu o salto deste vírus para a espécie humana.
Assuntos
Animais , Animais de Estimação/virologia , Síndrome Respiratória Aguda Grave/epidemiologia , Síndrome Respiratória Aguda Grave/transmissão , Síndrome Respiratória Aguda Grave/veterinária , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/patogenicidadeRESUMO
We studied the pathogenesis of Pseudocowpox virus (PCPV), a zoonotic parapoxvirus associated with mucocutaneous lesions in cattle. Inoculation of calves with PCPV isolate SD 76-65 intranasally (n = 6) or transdermally in the muzzle (n = 2) resulted in virus replication and shedding up to day 13 post-infection (pi). No local or systemic signs were observed in inoculated calves up to day 20pi, when the clinical monitoring was discontinued. However, from days 28-34 pi, seven (7/8) inoculated calves underwent an asynchronous clinical course characterized by development of a few (one or two) to countless papulo-pustular, erosive-fibrinous and scabby lesions in the muzzle, in some cases extending to the lips and gingiva. In some animals, the lesions coalesced, forming extensive fibrinotic/necrotic and scabby plaques covering almost entirely the muzzle. The clinical course lasted 8-15 days and spontaneously subsided after day 42pi. Infectious virus and/or viral DNA were detected in swabs collected from lesions of 5/8 animals between days 34 and 42pi. Histological examination of fragments collected from the muzzle lesions of two affected calves (day 36pi) revealed marked epidermal hyperplasia and severe orthokeratotic and parakeratotic hyperkeratosis, covered by thick scabs. The epidermis showed multifocal areas of keratinocyte coalescing necrosis and mild multifocal vacuolar degeneration. Sera of inoculated calves at 50pi showed partial virus neutralization at low dilutions, demonstrating seroconversion. The delayed and severe clinical course associated with virus persistence in lesions are novel findings and contribute for the understanding of PCPV pathogenesis.
Assuntos
Doenças dos Bovinos/virologia , Infecções por Poxviridae/veterinária , Vírus da Pseudovaríola das Vacas , Animais , Bovinos , Doenças dos Bovinos/patologia , Face/patologia , Infecções por Poxviridae/patologia , Infecções por Poxviridae/virologia , Carga Viral/veterináriaRESUMO
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 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 pestiviruses bovine viral diarrhea virus 1 (BVDV-1), 2 (BVDV-2), and HoBi-like (HoBiPeV) are endemic among Brazilian cattle, the world's largest commercial bovine herd. In the last two decades (1998-2018) over 300 bovine pestiviruses have been partially or fully sequenced in Brazil, including viruses from different regions, different epidemiological backgrounds, and associated with diverse clinical presentations. Phylogenetic analysis of these viruses demonstrated a predominance of BVDV-1 (54.4%), with subgenotypes -1a (33.9% of total) and -1b (16.3%) being more frequent and subgenotypes -1d, -1e, and -1i at very low frequencies. The overall BVDV-2 frequency was 25.7% but it varied largely by region, reaching up to 48% in Southern states. BVDV-2b was the predominant subgenotype (84.8% of BVDV-2), followed by BVDV-2a (8.86%). HoBiPeV accounted for 19.9% (61/307) of the genotyped viruses and were detected at high frequency in cattle from Northeastern states. These findings demonstrate a unique mix of pestivirus species and subgenotypes, unlike that seen in Europe or North America. The design of effective diagnostic tools, vaccines, and control programs for limiting bovine pestivirus infections in Brazil must take into consideration this unique mix of viruses. This article provides a critical review of two decades of genetic identification of pestiviruses in Brazil.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina/genética , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Brasil/epidemiologia , Bovinos , Vírus da Diarreia Viral Bovina/isolamento & purificação , GenótipoRESUMO
Hobi-like viruses comprise an unclassified group of bovine pestiviruses related to bovine viral diarrhea virus 1 (BVDV-1) and 2 (BVDV-2). These viruses were originally identified in fetal bovine serum from Brazilian origin and, subsequently, isolated from diseased animals in several countries. Herein we performed an antigenic characterization of eight Brazilian HoBi-like viruses isolated from persistently infected (PI) animals and from gastroenteric disease (2007-2015). Phylogenetic analysis based on the 5' unstranslated region (UTR) clustered these viruses with other HoBi-like viruses from European and Asiatic origin. Monoclonal antibody (MAb) binding indicated variability in the Hobi-like virus glycoprotein E2 and significant differences from the homologous BVDV-1 and BVDV-2 glycoprotein. Analysis of antigenic relatedness based on virus-neutralizing titers using virus-specific antisera revealed that HoBi-like viruses are antigenically very different from BVDV-1 and, to a lesser extent, from BVDV-2. Cross-neutralizing assays between pairs of HoBi-like viruses and their respective antisera indicated the existence of antigenic variability among these viruses, even for viruses isolated from the same herd in different occasions. Moreover, the identification of a HoBi-like isolate with low antigenic similarity with the other isolates indicates the potential existence of antigenic subgroups among HoBi-like virus isolates. Finally, sera of lambs immunized with commercial BVDV vaccines showed low or undetectable neutralizing activity against HoBi-like isolates. These results indicate significant antigenic differences between BVDV genotypes and Brazilian HoBi-like viruses and the existence of antigenic variability within this atypical group of pestiviruses. These findings extend the knowledge about the antigenic diversity of HoBi-like viruses and reinforce the need for their inclusion in current BVDV vaccines.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/sangue , Variação Antigênica , Doenças dos Bovinos/imunologia , Infecções por Pestivirus/veterinária , Pestivirus/imunologia , Animais , Bovinos , Doenças dos Bovinos/virologia , Vírus da Diarreia Viral Bovina Tipo 1/genética , Vírus da Diarreia Viral Bovina Tipo 1/imunologia , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Vírus da Diarreia Viral Bovina Tipo 2/genética , Vírus da Diarreia Viral Bovina Tipo 2/imunologia , Vírus da Diarreia Viral Bovina Tipo 2/isolamento & purificação , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/imunologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Imunização/veterinária , Pestivirus/genética , Pestivirus/isolamento & purificação , Infecções por Pestivirus/imunologia , Infecções por Pestivirus/virologia , Filogenia , OvinosRESUMO
Oncolytic virotherapy is a novel strategy for treatment of cancer in humans and companion animals as well. Canine distemper virus (CDV), a paramyxovirus, has proven to be oncolytic through induction of apoptosis in canine-derived tumour cells, yet the mechanism behind this inhibitory action is poorly understood. In this study, three human mammary tumour cell lines and one canine-derived adenofibrosarcoma cell line were tested regarding to their susceptibility to CDV infection, cell proliferation, apoptosis, mitochondrial membrane potential and expression of tumour necrosis factor-alpha-induced protein 8 (TNFAIP8). CDV replication-induced cytopathic effect, decrease of cell proliferation rates, and >45% of infected cells were considered death and/or under late apoptosis/necrosis. TNFAIP8 and CDVM gene expression were positively correlated in all cell lines. In addition, mitochondrial membrane depolarization was associated with increase in virus titres (p < 0.005). Thus, these results strongly suggest that both human and canine mammary tumour cells are potential candidates for studies concerning CDV-induced cancer therapy.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Vírus da Cinomose Canina/metabolismo , Terapia Viral Oncolítica/veterinária , Adenocarcinoma/terapia , Adenocarcinoma/veterinária , Animais , Apoptose , Neoplasias da Mama/terapia , Morte Celular , Linhagem Celular Tumoral , Proliferação de Células , Doenças do Cão/terapia , Cães , Feminino , Humanos , Neoplasias Mamárias Animais/terapia , Terapia Viral Oncolítica/métodosRESUMO
Pestivirus infections in ruminants result in significant economic losses worldwide. The aetiological agents are three species from the genus Pestivirus, family Flaviviridae, including bovine viral diarrhoea virus type 1 (BVDV-1), BVDV-2, border disease virus (BDV), and an atypical pestivirus named HoBi-like pestivirus. In this study, eighty-nine pestivirus isolates that were collected in Brazil between 1995 and 2014 and that originated from either cattle, fetal bovine serum (FBS) or as cell culture contaminants were genotyped based on a comparison of gene sequences from their 5' untranslated regions (5'UTR), N-terminal autoprotease (Npro ) and envelope glycoprotein 2 (E2). Of these isolates, 53.9% of the sequences were genotyped as BVDV-1, 33.7% as BVDV-2 and 12.4% as HoBi-like pestivirus. The prevalence of subgenotypes within the species was as follows: BVDV-1a (35.9%), BVDV-2b (31.4%), BVDV-1b (10.1%), BVDV-1d (6.7%), BVDV-2c (2.2%) and BVDV-1e (1.1%). BVDV-2c and BVDV-1e were detected for the first time in Brazil. This study revealed extensive genetic diversity among Brazilian pestivirus isolates, and the combination of pestiviruses that was detected is unique to Brazil. This information may serve as a foundation for designing and evaluating diagnostic tools and in the development of more effective vaccines; therefore, it may potentially contribute to pestivirus control and eradication.
Assuntos
Vírus da Diarreia Viral Bovina Tipo 1/genética , Vírus da Diarreia Viral Bovina Tipo 2/genética , Variação Genética , Animais , Brasil , Bovinos , FilogeniaRESUMO
In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two-dimensional (2D) and three-dimensional (3D) conditions. The 3D was performed in six-well AlgiMatrix™ (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50-125 µm for CC and 175-200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases-1, -2, -9 and -13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E-cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)-derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model.
Assuntos
Tecido Conjuntivo/metabolismo , Doenças do Cão/metabolismo , Neoplasias Mamárias Animais/metabolismo , Alicerces Teciduais , Animais , Caderinas/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular , Cães , Receptores ErbB/metabolismo , Feminino , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Peptídeos/metabolismoRESUMO
A bovine herpesvirus 1 (BoHV-1) defective in glycoprotein E (gE) was constructed from a Brazilian genital BoHV-1 isolate, by replacing the full gE coding region with the green fluorescent protein (GFP) gene for selection. Upon co-transfection of MDBK cells with genomic viral DNA plus the GFP-bearing gE-deletion plasmid, three fluorescent recombinant clones were obtained out of approximately 5000 viral plaques. Deletion of the gE gene and the presence of the GFP marker in the genome of recombinant viruses were confirmed by PCR. Despite forming smaller plaques, the BoHV-1â³gE recombinants replicated in MDBK cells with similar kinetics and to similar titers to that of the parental virus (SV56/90), demonstrating that the gE deletion had no deleterious effects on replication efficacy in vitro. Thirteen calves inoculated intramuscularly with BoHV-1â³gE developed virus neutralizing antibodies at day 42 post-infection (titers from 2 to 16), demonstrating the ability of the recombinant to replicate and to induce a serological response in vivo. Furthermore, the serological response induced by recombinant BoHV-1â³gE could be differentiated from that induced by wild-type BoHV-1 by the use of an anti-gE antibody ELISA kit. Taken together, these results indicated the potential application of recombinant BoHV-1 â³gE in vaccine formulations to prevent the losses caused by BoHV-1 infections while allowing for differentiation of vaccinated from naturally infected animals.
Assuntos
Deleção de Genes , Herpesvirus Bovino 1/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/imunologia , Animais , Bovinos , Eletroforese em Gel de Poliacrilamida , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 1/química , Herpesvirus Bovino 1/genética , Immunoblotting , Reação em Cadeia da Polimerase , Recombinação Genética/genética , Vacinas de Produtos Inativados/genética , Vacinas de Produtos Inativados/imunologia , Vacinas Virais/genéticaRESUMO
Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) share high genetic and antigenic similarities, but exhibit marked differences in tissue tropism and neurovirulence. The amino-terminal region of glycoprotein C (gC), which is markedly different in each of the viruses, is involved in virus binding to cellular receptors and in interactions with the immune system. This study investigated the genetic and antigenic differences of the 5′ region of the gC (5′ gC) gene (amino-terminal) of South American BoHV-1 (n=19) and BoHV-5 (n=25) isolates. Sequence alignments of 374 nucleotides (104 amino acids) revealed mean similarity levels of 97.3 and 94.2% among BoHV-1 gC (gC1), respectively, 96.8 and 95.6% among BoHV-5 gC (gC5), and 62 and 53.3% between gC1 and gC5. Differences included the absence of 40 amino acid residues (27 encompassing predicted linear epitopes) scattered throughout 5′ gC1 compared to 5′ gC5. Virus neutralizing assays testing BoHV-1 and BoHV-5 antisera against each isolate revealed a high degree of cross-neutralization between the viruses, yet some isolates were neutralized at very low titers by heterologous sera, and a few BoHV-5 isolates reacted weakly with either sera. The virus neutralization differences observed within the same viral species, and more pronounced between BoHV-1 and BoHV-5, likely reflect sequence differences in neutralizing epitopes. These results demonstrate that the 5′ gC region is well conserved within each viral species but is divergent between BoHV-1 and BoHV-5, likely contributing to their biological and antigenic differences.
Assuntos
Humanos , Anti-Infecciosos/uso terapêutico , Revisão de Uso de Medicamentos , Política Organizacional , Assistência Ambulatorial/organização & administração , Assistência Ambulatorial/normas , Pesquisa Biomédica , Resistência Microbiana a Medicamentos , Revisão de Uso de Medicamentos/legislação & jurisprudência , Revisão de Uso de Medicamentos/organização & administração , Revisão de Uso de Medicamentos/normas , Avaliação de Programas e Projetos de Saúde , Sociedades Médicas , Estados UnidosRESUMO
Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) share high genetic and antigenic similarities, but exhibit marked differences in tissue tropism and neurovirulence. The amino-terminal region of glycoprotein C (gC), which is markedly different in each of the viruses, is involved in virus binding to cellular receptors and in interactions with the immune system. This study investigated the genetic and antigenic differences of the 5' region of the gC (5' gC) gene (amino-terminal) of South American BoHV-1 (n=19) and BoHV-5 (n=25) isolates. Sequence alignments of 374 nucleotides (104 amino acids) revealed mean similarity levels of 97.3 and 94.2% among BoHV-1 gC (gC1), respectively, 96.8 and 95.6% among BoHV-5 gC (gC5), and 62 and 53.3% between gC1 and gC5. Differences included the absence of 40 amino acid residues (27 encompassing predicted linear epitopes) scattered throughout 5' gC1 compared to 5' gC5. Virus neutralizing assays testing BoHV-1 and BoHV-5 antisera against each isolate revealed a high degree of cross-neutralization between the viruses, yet some isolates were neutralized at very low titers by heterologous sera, and a few BoHV-5 isolates reacted weakly with either sera. The virus neutralization differences observed within the same viral species, and more pronounced between BoHV-1 and BoHV-5, likely reflect sequence differences in neutralizing epitopes. These results demonstrate that the 5' gC region is well conserved within each viral species but is divergent between BoHV-1 and BoHV-5, likely contributing to their biological and antigenic differences.
Assuntos
Região 5'-Flanqueadora/genética , DNA Viral/genética , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 5/genética , Análise de Sequência de DNA , Proteínas do Envelope Viral/genética , Animais , Antígenos Virais/análise , Bovinos , Epitopos/análise , Herpesvirus Bovino 1/imunologia , Herpesvirus Bovino 1/patogenicidade , Herpesvirus Bovino 5/imunologia , Herpesvirus Bovino 5/patogenicidade , Testes de Neutralização , Especificidade de Órgãos , Filogenia , Reação em Cadeia da Polimerase , Alinhamento de Sequência , América do Sul , VirulênciaRESUMO
The identification and elimination of persistently infected (PI) cattle are the most effective measures for controlling bovine pestiviruses, including bovine viral diarrhea virus (BVDV) and the emerging HoBi-like viruses. Here, colostrum-deprived calves persistently infected with HoBi-like pestivirus (HoBi-like PI calves) were generated and sampled (serum, buffy coat, and ear notches) on the day of birth (DOB) and weekly for 5 consecutive weeks. The samples were subjected to diagnostic tests for BVDV--two reverse transcriptase PCR (RT-PCR) assays, two commercial real-time RT quantitative PCR (RT-qPCR), two antigen capture enzyme-linked immunosorbent assays (ACE), and immunohistochemistry (IHC)--and to HoBi-like virus-specific RT-PCR and RT-qPCR assays. The rate of false negatives varied among the calves. The HoBi-like virus-specific RT-PCR detected HoBi-like virus in 83%, 75%, and 87% of the serum, buffy coat, and ear notch samples, respectively, while the HoBi-like RT-qPCR detected the virus in 83%, 96%, and 62%, respectively. In comparison, the BVDV RT-PCR test had a higher rate of false negatives in all tissue types, especially for the ear notch samples (missing detection in at least 68% of the samples). The commercial BVDV RT-qPCRs and IHC detected 100% of the ear notch samples as positive. While ACE based on the BVDV glycoprotein E(rns) detected infection in at least 87% of ear notches, no infections were detected using NS3-based ACE. The BVDV RT-qPCR, ACE, and IHC yielded higher levels of detection than the HoBi-like virus-specific assays, although the lack of differentiation between BVDV and HoBi-like viruses would make these tests of limited use for the control and/or surveillance of persistent HoBi-like virus infection. An improvement in HoBi-like virus tests is required before a reliable HoBi-like PI surveillance program can be designed.
Assuntos
Técnicas de Laboratório Clínico/métodos , Testes Diagnósticos de Rotina/métodos , Infecções por Pestivirus/veterinária , Pestivirus/isolamento & purificação , Animais , Buffy Coat/virologia , Bovinos , Orelha/virologia , Reações Falso-Negativas , Imunoensaio/métodos , Imuno-Histoquímica/métodos , Técnicas de Diagnóstico Molecular/métodos , Infecções por Pestivirus/diagnóstico , Soro/virologiaRESUMO
The immunostimulatory properties of inactivated Parapoxvirus ovis (iPPVO) have long been investigated in different animal species and experimental settings. In this study, we investigated the effects of iPPVO on cytokine expression in mice after intraperitoneal inoculation. Spleen and sera collected from iPPVO-treated mice at intervals after inoculation were submitted to cytokine mRNA determination by real-time PCR (qPCR), serum protein concentration by ELISA, and interferon (IFN)-α/ß activity by bioassay. The spleen of iPPVO-treated animals showed a significant increase in mRNA expression of all cytokines assayed, with different kinetics and magnitude. Proinflammatory cytokines interleukin (IL)-1ß, tumor necrosis factor-alpha (TNF-α), and IL-8 mRNA peaked at 24 hours postinoculation (hpi; 5.4-fold increase) and 48 hpi (3- and 10-fold increases), respectively. A 15-fold increase in IFN-γ and 6-fold IL-12 mRNA increase were detected at 48 and 24 hpi, respectively. Increased expression of autoregulatory cytokines (Th2), mainly IL-10 and IL-4, could be detected at later times (72 and 96 hpi) with peaks of 4.7- and 4.9-fold increases, respectively. IFN-I antiviral activity against encephalomyocarditis virus was demonstrated in sera of treated animals between 6 and 12 hpi, with a >90% reduction in the number of plaques. Measurement of serum proteins by ELISA revealed increased levels of IL-1, TNF-α, IL-12, IFN-γ, and IL-10, with kinetics similar to those observed by qPCR, especially for IL-12 and IFN-γ. These data demonstrate that iPPVO induced a transient and complex cytokine response, initially represented by Th1-related cytokines followed by autoregulatory and Th2 cytokines.
Assuntos
Citocinas/metabolismo , Vírus do Orf/imunologia , Células Th1/metabolismo , Animais , Citocinas/sangue , Citocinas/imunologia , DNA Complementar/biossíntese , Ensaio de Imunoadsorção Enzimática , Feminino , Camundongos , Parapoxvirus/imunologia , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Células Th1/virologia , Fatores de TempoRESUMO
The immunostimulatory properties of inactivated Parapoxvirus ovis (iPPVO) have long been investigated in different animal species and experimental settings. In this study, we investigated the effects of iPPVO on cytokine expression in mice after intraperitoneal inoculation. Spleen and sera collected from iPPVO-treated mice at intervals after inoculation were submitted to cytokine mRNA determination by real-time PCR (qPCR), serum protein concentration by ELISA, and interferon (IFN)-α/β activity by bioassay. The spleen of iPPVO-treated animals showed a significant increase in mRNA expression of all cytokines assayed, with different kinetics and magnitude. Proinflammatory cytokines interleukin (IL)-1β, tumor necrosis factor-alpha (TNF-α), and IL-8 mRNA peaked at 24 hours postinoculation (hpi; 5.4-fold increase) and 48 hpi (3- and 10-fold increases), respectively. A 15-fold increase in IFN-γ and 6-fold IL-12 mRNA increase were detected at 48 and 24 hpi, respectively. Increased expression of autoregulatory cytokines (Th2), mainly IL-10 and IL-4, could be detected at later times (72 and 96 hpi) with peaks of 4.7- and 4.9-fold increases, respectively. IFN-I antiviral activity against encephalomyocarditis virus was demonstrated in sera of treated animals between 6 and 12 hpi, with a >90% reduction in the number of plaques. Measurement of serum proteins by ELISA revealed increased levels of IL-1, TNF-α, IL-12, IFN-γ, and IL-10, with kinetics similar to those observed by qPCR, especially for IL-12 and IFN-γ. These data demonstrate that iPPVO induced a transient and complex cytokine response, initially represented by Th1-related cytokines followed by autoregulatory and Th2 cytokines.