Genome of a husavirus from Southern Brazil

ABSTRACT New viruses of the Picornavirales order have been discovered with the increase in the number of sequences obtained by high-throughput sequencing, as well as human stool-associated RNA virus (husavirus [HuV]), found in human stool samples. However, there is much to be clarified about HuV. Its cellular host, evolutionary history, and other biological characteristics are still unknown. Therefore, samples collected from human beings and environmental samples in a watershed in Southern Brazil were processed for the metagenomic library. Upon metagenomic analysis, we identified a HuV (husavirus LMM_67754 OP019707) genome with 8,846 bp, which was reported for the first time in Southern Brazil. The new genome presents only 37% of nucleotide identity with Brazilian strains and more than 90% with genomes from China, Vietnam, Venezuela, and the Netherlands. The HuV phylogeny presents significant differences among genomes, probably because multiple introductions of the virus may have occurred. Many questions still need to be answered about HuV. Therefore, more sequences and studies on this virus are necessary to improve the comprehension of the unknown origin of Picornavirales.

Chikungunya virus infection in the southernmost state of Brazil was characterised by self-limited transmission (2017-2019) and a larger 2021 outbreak

BACKGROUND Chikungunya is a mosquito-borne virus that has been causing large outbreaks in the Americas since 2014. In Brazil, Asian-Caribbean (AC) and East-Central-South-African (ECSA) genotypes have been detected and lead to large outbreaks in several Brazilian states. In Rio Grande do Sul (RS), the southernmost state of Brazil, the first cases were reported in 2016. OBJECTIVES AND METHODS We employed genome sequencing and epidemiological investigation to characterise the Chikungunya fever (CHIKF) burden in RS between 2017-2021. FINDINGS We detected an increasing CHIKF burden linked to travel associated introductions and communitary transmission of distinct lineages of the ECSA genotype during this period. MAIN CONCLUSIONS Until 2020, CHIKV introductions were most travel associated and transmission was limited. Then, in 2021, the largest outbreak occurred in the state associated with the introduction of a new ECSA lineage. CHIKV outbreaks are likely to occur in the near future due to abundant competent vectors and a susceptible population, exposing more than 11 million inhabitants to an increasing infection risk.

Temporal dynamics of human mastadenovirus species in cases of respiratory illness in southern Brazil

Human mastadenovirus (HAdV) genus is related to several diseases, among them upper and lower respiratory tract illness. HAdV species B, C, D, and E are mainly associated with respiratory infections. The goal of this work was to identify the HAdV species associated with respiratory infections in hospitalized patients from southern Brazil. Samples were collected from 1996 to 2004 and 2011 to 2017. During this period, 28,524 samples were collected, and 9983 were positive for respiratory viruses, being 435 for HAdV. From these 435 samples, 57 were selected for characterization of HAdV species. For screening the presence of HAdV, a partial sequence of the DNA polymerase gene (DNApol gene) was amplified by nested PCR. Partial nucleotide sequencing was performed in positive samples, and HAdV (DNApol gene) was detected in 53 samples: species B (28;49.1%), C (16;8.0%), D (2; 3.5%), E (5; 8.7%), and untyped (2; 3.5%). Specie D was found only in 2017 and specie E in 2011 and 2012. The age of the patients ranged from < 1 to 81 years old, and 62.3%were male. No relationship between gender orage and identified HAdV species were observed. In addition, in the period of 2013­2017, 18 samples from patients who died were analyzed: 11 were related to species B, 4 to C, and 2 to D and 1 remained untyped. Circulation of HAdV species D and Evaried over the years, but species B and C were present throughout the evaluated period. In addition, respiratory infections by HAdVaffect elderly and children mainly. (AU)

Seasonal variation on the presence of adenoviruses in stools from non-diarrheic patients

Human adenoviruses (HAdV), members of the Adenoviridae family, are excreted through the fecal route and may be present in the feces of humans consuming contaminated food or water. The presence of HAdV from different serotypes in the feces of healthy individuals was already reported using conventional polymerase chain reaction; however, real-time PCR (qPCR) may reveal not only the rates of detection as well as demonstrate the viral loads excreted by healthy persons. Aiming to identify and characterize the presence of adenoviruses in stool samples, 147 fecal samples from patients with no records of diarrhea were analyzed (74 from winter season and 73 from summer) by Real-Time PCR (qPCR) assay and conventional PCR. HAdV genome was present in 43.8% (32/73) of stools samples collected during summer season and 21.6% (16/74) during winter. The rate of detection of genomic copies (gc) ranged from 4.04×10² to 6.72×10⁵gc/g of feces among the 147 samples analyzed, of which the ranged of genomic copies of DNA HAdV was major in summer. All samples were negative when tested for rotaviruses (RV) and noroviruses (NoV) by PCR conventional and qPCR respectively. HAdV is excreted constantly by infected individuals in the absence of clinical signs and the occurrence may vary seasonally.

Quantitative vs conventional PCR for detectionof human adenoviruses in water and sediment samples / PCR quantitativa versus convencional para a detecção de adenovírus humano em amostras de água e sedimento

SUMMARYHuman Adenoviruses (HAdV) are notably resistant in the environment. These agents may serve as effective indicators of fecal contamination, and may act as causative agents of a number of different diseases in human beings. Conventional polymerase chain reaction (PCR) and, more recently, quantitative PCR (qPCR) are widely used for detection of viral agents in environmental matrices. In the present study PCR and SYBR(r)Green qPCR assays were compared for detection of HAdV in water (55) and sediments (20) samples of spring and artesian wells, ponds and streams, collected from dairy farms. By the quantitative methodology HAdV were detected in 87.3% of the water samples and 80% of the sediments, while by the conventional PCR 47.3% and 35% were detected in water samples and sediments, respectively.

RESUMOOs adenovírus humanos (HAdV) são notavelmente resistentes ao ambiente. Estes agentes podem servir como indicadores efetivos de contaminação fecal, tanto quanto podem atuar como agentes causadores de diferentes doenças em seres humanos. A reação em cadeia da polimerase (PCR) e mais recentemente a PCR quantitativa (qPCR) são amplamente usadas para detecção de agentes virais em matrizes ambientais. No presente estudo, PCR e SYBR(r)Green qPCR foram comparadas para a detecção de HAdV em amostras de água (55) e sedimento (20) provenientes de nascentes, poços, açudes e arroios coletadas em propriedades leiteiras. A metodologia quantitativa detectou HAdV em 87,3% das amostras de água e 80% dos sedimentos, enquanto por PCR convencional a detecção foi de 47,3% e 35%, respectivamente.

MULTIPLEX SYBR® GREEN-REAL TIME PCR (qPCR) ASSAY FOR THE DETECTION AND DIFFERENTIATION OF Bartonella henselae AND Bartonella clarridgeiae IN CATS / PCR em Tempo Real (qPCR) multiplex utilizando SYBR® Green para a detecção e diferenciação de Bartonella henselae e Bartonella clarridgeiae em gatos

A novel SYBR® green-real time polymerase chain reaction (qPCR) was developed to detect two Bartonella species, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.

Um novo teste baseado na reação em cadeia da polimerase em tempo real (qPCR) com SYBR ® Green foi desenvolvido para detectar duas espécies de Bartonella, B. henselae e B. clarridgeiae, diretamente em amostras de sangue. Este teste foi utilizado em amostras de sangue obtidas de gatos que vivem em abrigos de animais do sul do Brasil. Os resultados foram comparados aos obtidos pelo PCR convencional utilizado para a detecção de Bartonella spp. Das 47 amostras analisadas, oito foram positivas no PCR convencional e 12 foram positivas para qPCR. A reação de qPCR, permitiu a detecção da presença simultânea de B. henselae e B. clarridgeiae em duas destas amostras. Os resultados mostram que a qPCR aqui descrita pode ser uma ferramenta confiável para a detecção e diferenciação de duas espécies importantes de Bartonella spp.

Seroprevalence of Hepatitis B and C markers at the population level in the municipality of Caxias do Sul, southern Brazil

Chronic viral hepatitis are main public health problems worldwide. Data about the seroprevalence to Hepatitis B and C viruses (HBV e HCV) at the population level are scarce on Brazil and especially for the Southern region of the country. The seroprevalence to HBV and HBC antigens was evaluated on a large portion of the population of the municipality of Caxias do Sul (427,858 inhabitants), Brazil. A total of 60,604 individual serum samples collected from 2008 to 2011 were screened for HBV surface antigen (HBsAg) and for antibodies against HBsAg; anti HCV antibodies were measured by ELISA (Enzyme Linked Immunosorbent Assay). Overall, 1.63% of the individuals were positive for HBsAg and 1.43% showed seropositivity to HCV. From the total, 31,749 samples were analyzed for HBsAg and 28,855 for HCV. For HBsAg, 519 samples showed positive (1.63%) while the results for 37 patients (0.12%) remained inconclusive. For the anti-HCV test, 412 individuals (1.43%) showed positive. From the positive samples for HBsAg 216 (50.6%) were from male individuals whereas for anti-HCV the seroprevalence was slightly higher for females 216 (52.4%). The higher prevalence for both hepatitis viruses were found among individuals at the age group of 40 to 59 years and the lower levels of positivity for both HBV and HBC were among children and teenagers.

Emerging animal viruses: real threats or simple bystanders? / Vírus emergentes de animais: verdadeiras ameaças ou meros espectadores?

The list of animal viruses has been frequently added of new members raising permanent concerns to virologists and veterinarians. The pathogenic potential and association with disease have been clearly demonstrated for some, but not for all of these emerging viruses. This review describes recent discoveries of animal viruses and their potential relevance for veterinary practice. Dogs were considered refractory to influenza viruses until 2004, when an influenza A virus subtype H3N8 was transmitted from horses and produced severe respiratory disease in racing greyhounds in Florida/USA. The novel virus, named canine influenza virus (CIV), is considered now a separate virus lineage and has spread among urban canine population in the USA. A new pestivirus (Flaviviridae), tentatively called HoBi-like pestivirus, was identified in 2004 in commercial fetal bovine serum from Brazil. Hobi-like viruses are genetically and antigenically related to bovine viral diarrhea virus (BVDV) and induce similar clinical manifestations. These novel viruses seem to be widespread in Brazilian herds and have also been detected in Southeast Asia and Europe. In 2011, a novel mosquito-borne orthobunyavirus, named Schmallenberg virus (SBV), was associated with fever, drop in milk production, abortion and newborn malformation in cattle and sheep in Germany. Subsequently, the virus disseminated over several European countries and currently represents a real treat for animal health. [...] Finally, the long time and intensive search for animal relatives of human hepatitis C virus (HCV) has led to the identification of novel hepaciviruses in dogs (canine hepacivirus [CHV]), horses (non-primate hepaciviruses [NPHV] or Theiler's disease associated virus [TDAV]) and rodents. For these, a clear and definitive association with disease is still lacking and only time and investigation will tell whether they are real disease agents or simple spectators.

O número de vírus animais cresce continuamente, causando preocupação permanente a virologistas e veterinários. O potencial patogênico e associação com doença tem sido claramente demonstrado para alguns - mas não para todos - vírus emergentes. Esse artigo apresenta uma breve revisão das recentes descobertas de vírus animais e a sua potencial relevância para saúde animal. Cães eram considerados refratários aos vírus da influenza até 2004, quando um vírus influenza A subtipo H3N8 foi transmitido de equinos e causou doença respiratória severa em cães galgos na Flórida/EUA. O novo vírus, denominado vírus da influenza canina (CIV), agora considerado uma linhagem distinta do vírus da influenza equina, disseminou-se na população canina urbana dos EUA. Um novo Pestivirus (Flaviviridae) - provisoriamente denominado pestivírus Hobi-like - foi identificado em 2004 em soro fetal bovino importado do Brasil. Os vírus Hobi-like são genética e antigenicamente relacionados com o vírus da diarreia viral bovina (BVDV) e induzem manifestações clínicas semelhantes. A sua origem e distribuição são desconhecidas, mas estão aparentemente disseminados no rebanho brasileiro e já foram identificados no sudeste asiático e na Europa. Em 2011, um novo buniavírus transmitido por mosquitos, denominado vírus Schmallemberg (SBV), foi associado com febre, redução da produção de leite, abortos e malformações fetais em bovinos e ovinos da Alemanha. [...] Finalmente, a longa e intensiva busca por vírus animais relacionados ao vírus da hepatite C humana (HCV) tem levado a identificação de "novos" pestivírus em cães (canine hepacivirus [CHV]), equinos (hepacivirus de não-primatas [NPHV] ou vírus associado à doença de Theiler [TDAV]) e em roedores. Para estes, uma associação clara e definitiva com doença ainda não foi demonstrada e apenas tempo e investigação irão dizer se são patógenos reais ou apenas espectadores.

Detection of human adenovirus, rotavirus and enterovirus in water samples collected on dairy farms from Tenente Portela, Northwest of Rio Grande do Sul, Brazil

Viral gastroenteritis and other waterborne diseases are a major concern for health in Brazil. A number of studies were conducted about the presence of viruses on water samples from Brazilian areas. However, the knowledge about the occurrence of viral contamination of drinking water sources in rural settings of the country is insufficient. On the present work, 15 samples from 5 dairy farms located at the municipality of Tenente Portela were collected and analysed for the presence of human adenoviruses (HAdV), as well as human enteroviruses (EV) and rotaviruses (RV). HAdV was present on 66.66% of the water samples, and have been found in all samples from artesian wells and springs, which are used as sources of drinking water for the individuals inhabiting those farms. EV and RV found only in one sample each. The detection rates of HAdV on the water from these dairy farms are alarming and point towards a situation of elevated environmental contamination by fecal microorganisms of human origin and poor basic sanitation conditions.

Detecção molecular de vírus da bronquite infecciosa em plantéis de avós, matrizes e frangos de corte no Rio Grande do Sul e Mato Grosso / Molecular detection of infectious bronchitis virus in flocks of grandmothers, mothers and broiler chicken in Rio Grande do Sul and Mato Grosso

O vírus da Bronquite Infecciosa das galinhas (VBI) pertence ao grupo 3 da família Coronaviridae e é o causador de desordens respiratórias e renais em frangos de corte. A vacinação com vacinas vivas é praticada em matrizes e avós e muitas vezes também nos plantéis destinados ao abate. As vacinas utilizadas no Brasil são usualmente do sorogrupo Massachusetts e baseadas nas amostras H120 e H52. É comum que após a vacinação o vírus vacinal seja detectado por isolamento em ovos embrionados ou por métodos moleculares por até 4 semanas. Após essa data, normalmente, não há detecção de vírus e o VBI, quando encontrado, pode representar recirculação do vírus vacinal no plantel ou a introdução de uma nova cepa do vírus. No presente estudo, para avaliar a circulação do vírus em plantéis de frangos e reprodutoras nos estados do Rio Grande do Sul e Mato Grosso do Sul, foram coletadas 240 traqueias e rins de aves de 48 plantéis, sendo (20 exemplares/4 plantéis) de avós, (80 exemplares/16 plantéis) de matrizes e (140 exemplares/28 plantéis) de frangos de corte, as quais foram analisadas em misturas de cinco amostras. Todos os animais eram vacinados e as amostras foram coletadas ao redor de 2 a 48 semanas após a vacinação. A presença de VBI foi determinada com auxílio de uma reação em cadeia da polimerase tipo nested, direcionada ao gene da proteína S1, padronizada neste estudo. Das 48 amostras testadas, 14 resultaram positivas: cinco foram oriundas de aves vacinadas há menos de quatro semanas na data da coleta e nove eram de amostras de aves vacinadas há mais de quatro semanas, o que pode ser devido à recirculação do vírus vacinal ou mesmo introdução de vírus selvagem nos plantéis.

Infectious bronchitis virus (IBV, Avian Coronavirus) from chickens belongs to group 3 of the family Coronaviridae and causes respiratory and renal disorders in broilers. Vaccination using live vaccines is generally performed in mothers and grandmothers, as well as often in flocks for slaughter. The vaccines used in Brazil are usually from serogroup Massachusetts and based on standard samples of the virus at passages H120 and H52. It is common that after vaccination the vaccine virus is detected by isolation in embryonated eggs or by molecular methods for up to four weeks. After, there is usually no virus detection and any IBV found may represent recirculation of the vaccine virus in the flock or the introduction of a new strain. In this study, to evaluate the circulation of the virus in poultry flocks and breeders in the state of Rio Grande do Sul and Mato Grosso do Sul, 240 samples were collected from tracheas and kidneys of birds from 48 flocks, and (20 biological samples / 4 flocks) from grandmothers (80 samples/16 flocks) and mothers (140 samples/28 flocks) from broilers, which were analyzed in pools of five samples. All animals were vaccinated and samples were collected around 2-48 weeks after vaccination. The presence of IBV was determined with the aid of a polymerase chain reaction "nested" gene-directed protein S1, standardized in this study. From the 48 samples tested, 14 were positive: 5 were from birds vaccinated after less than 4 weeks and 9 were from birds vaccinated more than four weeks should be wild viruses or represent the recirculation of the vaccine virus.

Perfil dos bolsistas de produtividade do Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) na área de Medicina Veterinária / Profile of the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) productivity fellows in the area of Veterinary Medicine

In this study, data regarding the population of productivity fellows from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Committee of Veterinary Medicine, were evaluated by calculation of scientometrical indexes as an effort to examine the profile of researchers from different levels on these parameters. Other variables, such as the place of doctorate studies, gender, institution, and advisory of human resources were also recorded and evaluated. There is a clear predominance of men (72.37%), which are mostly settled in the South and Southeast. Most of the recipients were awarded his doctorate in Brazil. From the analyzed parameters, advisory of human resources is very high at all levels; among the scientometric variables, the citations indexes and h-index are decreasing from the highest level (PQ-1A) to level 2, yet the intragroup variation is very high for these and other derivatives of the H-index. A modification of the h-index, the AWCR index, in which the calculation takes into account the age of articles published and cited, seems more appropriate to stratify the scholarship of fellows. Studies like these could be repeated in the medium term in order to improve the ranking formulas of scholars.

No presente estudo, submeteram-se dados da população de bolsistas de produtividade do Comitê de Medicina Veterinária do Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) ao cálculo de índices cientométricos, em um esforço para avaliar o perfil de pesquisadores de diferentes níveis perante vários parâmetros. Outras variáveis, como o local de formação, gênero, local de trabalho e formação de recursos humanos foram também anotados e avaliados. Há uma clara predominância do gênero masculino (72,37%), os quais se encontram em sua grande maioria radicados nas regiões Sul e Sudeste do país. A maioria dos bolsistas concluiu seu doutorado no Brasil. Dentre os parâmetros de produção analisados, a formação de recursos humanos é alta em todos os níveis; dentre as variáveis cientométricas, os índices de citações e o índice h são decrescentes do nível mais alto (PQ-1A) ao nível 2; todavia, a variação intragrupos é muito alta para estes e outros derivado do índice h. Uma modificação do índice H, o índice AWCR, em que o cálculo leva em conta a idade dos artigos publicados e citados, parece mais adequado à estratificação dos bolsistas. No âmbito geral, estudos deste tipo poderiam ser repetidos no médio prazo com vistas a aprimorar as fórmulas de ranqueamento de bolsistas.

Analysis of isotype-specific antibody responses to bovine herpesviruses 1.1 and 1.2a allows to estimate the stage of infection

Specific IgM, IgA, IgG1, IgG2, as well as neutralizing antibody responses were evaluated in sera of calves experimentally infected with two isolates of bovine herpesvirus type 1 (BoHV1) of distinct subtypes (subtype 1, BoHV1.1; subtype 2a, BoHV-1.2a). No significant differences were observed in the antibody responses induced by each BoHV-1 subtype. The antibody responses following primary acute infection were characterized by an increase in specific IgM and IgA levels between days 2 and 14 post inoculation (pi). IgG1 was detected from days 11 to 30 pi. IgG2 was detected on the sample taken on day 30 pi. Reactivation of infection following dexamethasone administration induced a significant rise in IgA levels, whereas IgG1 and IgG2 levels, which were at high levels from the beginning of the reactivation process, showed a slight alteration after corticosteroid treatment. These results suggest that it is possible to estimate the dynamics of BoHV-1 infections with basis on the analysis of class- and subclass-specific antibody responses. Such information may be particularly useful for the study of the kinetics of the infection in a herd and to aid in the adoption of appropriate control measures..

Detecção molecular e análise filogenética do gene H de amostras do vírus da cinomose canina em circulação no município de Campinas, São Paulo / Molecular detection and phylogenetic analysis of the gene H from canine distemper virus isolates circulating at the municipality of Campinas, São Paulo

O vírus da cinomose canina (CDV), um Morbillivirus da família Paramyxoviridae, é o agente etiológico de doença neurológica e sistêmica em cães. O diagnóstico laboratorial da infecção requer o isolamento viral ou detecção do material genético do vírus em secreções ou tecidos de cães com suspeita clínica da doença. A diversidade genética entre os isolados de CDV pode ser aferida pelo sequenciamento efilogenia molecular do gene que codifica a hemaglutinina viral (gene H), havendo atualmente um especial interesse em comparar as amostras circulantes a campo com o genogrupo América-1, que abrange as cepas presentes nas vacinas disponíveis no mercado. No presente estudo, foi realizada a detecção molecular do gene H de CDV a partir de amostras biológicas colhidas ante- e post- -mortem de 15 cães com sinais clínicos sugestivos de cinomose na região metropolitana de Campinas, São Paulo. Dez dos 15 cães analisados tiveram ao menos um órgão positivo na detecção molecular e os amplicons obtidos foram submetidos ao sequenciamento nucleotídico seguido de análise filogenética molecular. De forma semelhante ao que já foi reportado para estudo analisando a diversidade do gene H em outros países, a reconstrução filogenética obtida para as amostras de casos de cinomose da região de Campinas demonstrou as mesmas foram agrupadas junto a amostras norte-americanas, europeias e japonesas recentes, em um grupo genético distinto do grupo de amostras clássicas de CDV, nomeado America-1, o qual engloba as estirpes vacinais Snyder Hill, Onderstepoort e Lederle.

Canine distemper virus (CDV), a Morbillivirus of the family Paramyxoviridae, is the etiological agent of neurological and systemic disease in dogs. The laboratory diagnosis of infection requires viral isolation or detection of genetic material of the virus in secretions or tissues of dogs with clinical suspicion of the disease. The genetic diversity among isolates of CDV can be assessed by sequencing and phylogenetic analysis of the gene that encodes the viral hemagglutinin (H gene), and there is currently a special interest in comparing the strains currently circulating in the field with the genogroup America-1, which comprises strains present in vaccines available in the market. In this study, the molecular detection of CDV gene H was performed from biological samples harvested ante-and post-mortem from 15 dogs with clinical signs suggestive of canine distemper in the metropolitan region of Campinas, São Paulo. Ten of the 15 dogs examined had at least one positive organ under molecular detection and the obtained amplicons were sequenced and further analyzed by molecular phylogenetic analysis. Similarly to what has already been reported on previous studies regarding the diversity of the gene H in other countries, the phylogenetic reconstruction obtained for the samples of cases of distemper from Campinas region showed they were grouped with the North American, European and Japanese newly described samples, a genetic group distinguished from classical samples of CDV, named America-1, which encompasses the vaccine strains Snyder Hill, Onderstepoort and Lederle.

Phylogenetic characterization of bovine parainfluenza 3 from contaminated cell cultures and field isolates from Brazil

Genomic fragments of the HN and L genes from Brazilian bovine parainfluenza 3 virus (bPIV-3) isolated as contaminants from cell cultures and clinical specimens were amplified by reverse transcription-polymerase chain reaction (RT-PCR), sequenced using specific degenerate primers and analyzed by phylogenetic comparison with reference strains of bPI3V. The Brazilian isolates revealed a high degree of genomic when compared to SF4/32 prototype strain, within the recently proposed genotype A of bPIV-3.

Detecção molecular e análise filogenética de vírus respiratório sincicial bovino (BRSV) em swabs e tecido pulmonar de bovinos adultos / Molecular detection and phylogenetic analysis of bovine respiratory syncytial virus (BRSV) in swabs and lung tissues of adult cattle

Bovine respiratory syncytial viruses virus (BRSV) is one of the etiologic agents of pneumonia in young cattle. Few studies have been made aiming detection of the virus in samples collected from adult animals, especially those asymptomatic bovines. However, it is assumed that infections in these groups may occur mostly asymptomatic and this would be an important mechanism for maintaining of BRSV in herds. In this study, the goal was to conduct an analysis of the occurrence of asymptomatic infections by BRSV in lung samples (n=68) and nasal swabs (209) taken from adult animals collected in abattoirs from Southern and Southeastern Brazil respectively, to detect via polymerase chain reaction the occurrence of infected animals in populations of adult cattle. The samples that resulted positive (6) on RT-PCR were subsequently subjected to cutting with restriction enzymes and sequencing for genetic characterization (2 samples). All samples belongs to subgroup B of BRSV, which is reported as the one circulating in Brazil. The results obtained demonstrate that BRSV may be present in samples taken from adult animals, which is in agreement the hypothesis that infections in adults run in a sub-clinical way that may be of importance as a maintenance mechanism of the virus in bovine herds.

O vírus respiratório sincicial bovino (BRSV) é rebanhos. No presente estudo, o objetivo foi realizar uma um dos agentes etiológicos de pneumonias em bovinos jo-análise da prevalência de infecções assintomáticas pelo vens. Poucos estudos foram realizados visando à detecção BRSV em pulmões (n=68) e swabs nasais (209) coletados do agente em amostras coletadas de animais adultos, e em de bovinos adultos coletadas em frigoríficos da região Sul especial de bovinos assintomáticos. No entanto, presume-e Sudeste respectivamente, no sentido de detectar por in-se que as infecções ocorridas nestes grupos possam ocor-termédio de reação da polimerase em cadeia qual a taxa rer em sua maioria de forma assintomática e este seria de animais infectados em populações de animais adultos um mecanismo importante para manutenção do BRSV nos onde não ocorram sinais clínicos da infecção. As amostras positivas à RT-PCR (6) foram posteriormente submetidas ao corte com enzimas de restrição (REA) e sequenciamento para caracterização genética do gene F (2 das amostras). Todas as amostras se enquadram no subgrupo B de BRSV, o grupo circulante no Brasil conforme estudos anteriores. Os resultados obtidos demonstram que o BRSV pode estar presente em amostras obtidas de animais sadios, reforçando a hipótese de que infecções subclínicasfazem parte do mecanismo de manutenção do vírus nos rebanhos.

Vírus respiratórios e ventilação mecânica em lactentes brasileiros / Respiratory virus and mechanical ventilation in Brazilian infants

Introdução: Entre lactentes, Vírus Sincicial Respiratório e Metapneumovírus são agentes importantes de infecção respiratória baixa com necessidade de ventilação mecânica. Índice de Ventilação e Relação PaO2/FiO2 podem ser fatores prognósticos do tempo de ventilação mecânica nestes casos. Métodos: Dentre 284 lactentes (zero a 12 meses), hospitalizados por infecção respiratória aguda baixa em 2004, 2007 e 2008, foram avaliados 20 que necessitaram de ventilação mecânica. Análise da secreção nas ofaríngea para vírus por Polimerase Chain Reaction foi realizada; o Índice de Ventilação Mecânica e a Relação PaO2/FiO2 foram obtidos nos primeiros cinco dias de ventilação mecânica; tempo prolongado de ventilação pulmonar mecânica foi considerado sete dias ou mais. Resultados: Dez em vinte lactentes foram identificados com Vírus Sincial Respiratório; 0/20 foram positivos para Metapneumovírus. A análise estatística comparativa não mostrou diferenças entre Índice de Ventilação Mecânica e Relação PaO2/FiO2 e tempo de ventilação pulmonar prolongada entre os grupos Vírus Sincicial Respiratório positivo e negativo. A identificação do genótipo foi realizada em 6 de 10 Vírus Sincicial Respiratórios encontrados; o pequeno número de casos não permitiu relacionar a apresentação clínica com as características virais (subgrupos e genótipos). Conclusão: Índice de Ventilação Mecânica e Relação PaO2/FiO2 não foram úteis como fatores prognósticos de tempo de ventilação mecânica prolongada para este grupo. De maneira ideal, estudo com maior número de lactentes, teste para vários vírus, e testes para a imunidade inata e adaptativa, poderia mostrar o impacto destes fatores na evolução dos lactentes em ventilação pulmonar mecânica. Infelizmente, recursos para pesquisas como esta não estão facilmente disponíveis.

Background: Among infants, respiratory syncytial virus (RSV) and human metapneumovirus (HMPV) are important agents of lower respiratory tract infection requiring mechanical ventilation. Ventilation index and PaO2/FiO2 ratio may be prognostic factors for duration of mechanical ventilation in these cases. Methods: From a population of 284 infants (aged zero to 12 months) hospitalized with lower respiratory tract infection in 2004, 2007, and 2008, 20 infants requiring mechanical ventilation were evaluated. Nasopharyngeal secretions were analyzed for virus detection using a polymerase chain reaction (PCR) test. Ventilation index and PaO2/FiO2 ratio were obtained during the first five days of mechanical ventilation. Prolonged mechanical ventilation was defined as required ventilatory support for 7 days or longer. Results: Out of 20 infants assessed, 10 were positive for RSV and none for HMPV. Comparative statistical analysis showed no difference in ventilation index, PaO2/FiO2 ratio, and prolonged mechanical ventilation between RSV-positive and –negative groups. Genotypic identification was performed in 6 of 10 RSV-positive infants. The small number of cases did not allow a statistical correlation between clinical presentation and viral characteristics (subgroups and genotypes). Conclusion: Ventilation index and PaO2/FiO2 were not useful as prognostic factors for prolonged mechanical ventilation in this group. Ideally, studies involving more infants and including tests for several viruses and for innate and adaptive immunity should be conducted to further evaluate the impact of these factors on the outcome of infants requiring mechanical ventilation. Unfortunately, resources for this type of research are not readily available.

Antibody responses in mice after immunization with inactivated bovine respiratory syncytial virus using different adjuvants / Respostas de anticorpos em camundongos após imunização com vírus respiratório sincicial bovino inativado utilizando diferentes adjuvantes

Bovine respiratory syncytial virus (BRSV) causes pneumonia in young cattle. Modified-live-virus (MLV) and inactivated vaccines are currently used for the control of clinical effects of BRSV infections in cattle. On the present research, the stimulation of specific anti-BRSV immunoglobulin isotypes was investigated, through the use of different commercially available adjuvants (Water-in-oil emulsion, Quil A, Aluminum-hydroxide) in inbred mice (Balb/C and C57BL/6). BRSV antibodies were measured using an enzyme-linked immunosorbent assay (ELISA) and the results were compared to the antibody levels induced by immunization of animals using live-BRSV-virus. Water-in-oil emulsion and alum- adjuvant preparations induced higher levels of IgG1 immunoglobulins, whereas Quil A favored the production of IgG2 antibodies, this last being a more appropriate response profile for the specific case of BRSV. Not using adjuvants resulted in poor levels of specific antibodies. The isotype profile of specific antibodies obtained varied greatly depending on the adjuvants used. This information may be useful for the formulation of more effective BRSV inactivated vaccines; however, these findings have to be confirmed in cattle.

O vírus respiratório sincicial bovino (BRSV) causa pneumonia em bovinos jovens. Vacinas de vírus vivo modificado (MLV) e vacinas inativadas são atualmente utilizadas para o controle dos efeitos clínicos de infecções pelo BRSV em bovinos. No presente trabalho, investigou-se a estimulação dos isotipos de imunoglobulinas específicas anti-BRSV, através da utilização de diferentes adjuvantes disponíveis comercialmente (água em óleo de emulsão, Quil A, hidróxido de alumínio) em camundongos isogênicos (Balb/C e C57BL/6). Anticorpos contra o BRSV foram medidos usando-se um ensaio imunoenzimático (ELISA), e os resultados foram comparados com os níveis de anticorpos induzidos pela imunização de animais utilizando-se o BRSV vivo. As preparações em que se empregou óleo mineral e alumínio como adjuvantes induziram altos níveis de imunoglobulinas IgG1, enquanto QuilA favoreceu a produção de anticorpos de classe IgG2, sendo este último um perfil de resposta mais desejável para o caso específico de BRSV. A não utilização de adjuvantes resultou em baixa produção de anticorpos específicos. O perfil de isotipos de imunoglobulinas secretados variou bastante conforme o adjuvante utilizado. Esta informação pode ser útil futuramente na formulação de vacinas inativadas mais eficazes contra o BRSV. Todavia, esses achados devem ser confirmados em bovinos.

Prevalence of Bartonella henselae and Bartonella clarridgeiae in cats in the south of Brazil: a molecular study

Bartonella spp are the causative agent of cat scratch disease in humans. Cats are the natural reservoir of these bacteria and may infect humans through scratches, bites or fleas. Blood samples from 47 cats aged up to 12 months were collected for this study. All animals were lodged in municipal animal shelters in the Vale do Sinos region, Rio Grande do Sul, Brazil. Bartonella spp were detected by genus-specific polymerase chain reaction (PCR) and when the PCR was positive, the species were determined by DNA sequencing. A Giemsa-stained blood smear was also examined for the presence of intraerythrocytic elements suggestive of Bartonella spp infection. Phylogenetic analysis was also performed for all positive samples. Using molecular detection methods, Bartonella spp were detected in 17.02 percent (8/47) of the samples. In seven out of eight samples confirmed to be positive for Bartonella spp, blood smear examination revealed the presence of intraerythrocytic elements suggestive of Bartonella spp. Phylogenetic analysis characterized positive samples as Bartonella henselae (5) or Bartonella clarridgeiae (3). To the best of our knowledge, this is the first molecular study demonstrating the presence of Bartonella spp in cats from the Southern Region of Brazil.

Experimental infection of rabbits with a recombinant bovine herpesvirus type 5 (BoHV-5) gI, gE and US9-negative / Infecção experimental de coelhos com um recombinante do herpesvírus bovino tipo 5 defectivo na gI, gE e US9

Bovine herpesvirus type 5 (BoHV-5) is a major cause of viral meningoencephalitis in cattle. The expression of different viral proteins has been associated with BoHV-5 neuropathogenesis. Among these, gI, gE and US9 have been considered essential for the production of neurological disease in infected animals. To evaluate the role of gI, gE and US9 in neurovirulence, a recombinant from which the respective genes were deleted (BoHV-5 gI-/gE-/US9-) was constructed and inoculated in rabbits of two age groups (four and eight weeks-old). When the recombinant virus was inoculated through the paranasal sinuses of four weeks-old rabbits, neurological disease was observed and death was the outcome in 4 out of 13 (30.7 percent) animals, whereas clinical signs and death were observed in 11/13 (84.6 percent) of rabbits infected with the parental virus. In eight weeks-old rabbits, the BoHV-5 gI-/gE-/US9- did not induce clinically apparent disease and could not be reactivated after dexamethasone administration, whereas wild type BoHV-5 caused disease in 55.5 percent of the animals and was reactivated. These findings reveal that the simultaneous deletion of gI, gE and US9 genes did reduce but did not completely abolish the neurovirulence of BoHV-5 in rabbits, indicating that other viral genes may also play a role in the induction of neurological disease.

O herpesvírus bovino tipo 5 é uma das principais causas de meningoencefalite viral em bovinos. A expressão de diferentes proteínas virais tem sido associada à neuropatogenia do BoHV-5. Entre estas, a gI, gE e US9 têm sido consideradas essenciais para a indução de sinais neurológicos nos animais infectados. Para avaliar o papel das proteínas gI, gE e US9 na neurovirulência, construiu-se um recombinante no qual os genes que codificam estas proteínas foram deletados, denominado BoHV-5 gI-/gE-/US9-. Este vírus foi inoculado em coelhos de idades diferentes (quatro e oito semanas de idade). Quando o vírus recombinante foi inoculado nos seios paranasais de coelhos de quatro semanas de idade, doença neurológica e morte foram observadas em 4 dos 13 (30,7 por cento) animais, enquanto que sinais clínicos e morte foram observados em 11/13 (84,6 por cento) dos coelhos infectados com o vírus parental. Em coelhos de oito semanas de idade, o BoHV-5 gI-/gE-/US9- não induziu sinais clínicos aparentes e, após tentativa de reativação viral por tratamento com dexametasona, o vírus não foi re-excretado. Por outro lado, o vírus selvagem causou doença clínica em 55,5 por cento dos coelhos e foi re-excretado após tratamento com dexametasona. Estes achados revelam que a deleção simultânea dos genes gI, gE e US9 reduziu mas não aboliu completamente a neurovirulência do BoHV-5 em coelhos, indicando que outros genes virais possam ter papel na indução da doença neurológica.

Torque teno vírus (TTV) e poluição fecal / Torque teno virus (TTV) and fecal pollution

A pesquisa de agentes virais em amostras de água coletadas no ambiente teve seu início há aproximadamente sessenta anos. Inicialmente, o enfoque dos trabalhos estava voltado para a detecção dos enterovírus. Posteriormente outros vírus, especialmente entéricos com relevante impacto em saúde pública, tornaram-se alvo de pesquisa quanto à sua presença em amostras de água. Os agentes virais, normalmente associados a gastroenterites e hepatites, têm como principais características excreção em altas quantidades nas fezes de indivíduos infectados, elevada resistência à inativação no ambiente e ausência de replicação fora do hospedeiro. Tais características biológicas os tornam excelentes marcadores de contaminação ambiental oriunda da colonização de novas áreas pelo ser humano e podem atestar falhas nas estratégias de saneamento básico. Mais recentemente, o Torque Teno vírus (TTV) vem sendo considerado um candidato em potencial a agente biológico marcador no estudo de contaminação fecal da água por apresentar as características anteriormente mencionadas para os vírus entéricos e estar amplamente disseminado na população humana.