Eliana De Stefano

Ocorrência de anticorpos anti-Neospora caninum em cães da microrregião da Serra de Botucatu, Estado de São Paulo, Brasil

Neosporosis is a parasitic disease caused by a protozoon Neospora caninum, which is recognized as an important cause of bovine abortion and canine neuropathy. Considering the isolation of N. caninum and the frequent serology in bovines of our region, the objectives of the present trial were to evaluate the occurrence of antibodies anti-N. caninum in dogs of the Botucatu range micro region, in the State of Sao Paulo, and its association with gender, age and origin (urban area--exclusively from the city; rural area--only farms and cottages; and periurban area--access to the urban and rural areas) of the dogs studied. A total of 963 dogs of defined breeds or non-defined breeds, of both genders and different ages, showing no clinical symptoms, were analyzed. Animals were randomly selected during the anti-rabies vaccination campaign in the region, from May to September 1998. Serum samples obtained from the animals were evaluated by means of the Indirect Fluorescent Antibody Test (IFAT) using as antigen N. caninum, standard strain NC-1. A total of 245 animals were reagent (25.4% positive results), with 161 (27.5%) males and 84 (22.3%) females. According to the origin--urban, rural and periurban areas--223 (25.8%), 11(16.9%) and 11(33.3%) dogs were, respectively, reagent to the IFAT. All of the 11 cities in this region presented seropositive dogs with occurrence rates ranging from 53.5 to 8.9%. The lower positive percentage was observed in dogs below 1 year old (16.2%) when compared with animals from 1 to 4 years old and more than 4 years old (28.4% and 28.0% respectively), which did not present any differences between them. Results obtained characterized the seropositivity for N. caninum in dogs from all Municipalities in the Botucatu micro region showing the wide distribution of the agent in the region.

Erradicação do herpesvírus bovino - 1 (BHV-1) de um rebanho bovino leiteiro em manejo semi-intensivo

To eradicate the BHV-1 from a dairy cattle herd without vaccination, a preliminary serological survey was performed in 154 animals, that revealed 15.6% seropositives for BHV-1. The test used was serum neutralization with the microtitration technique. The seropositive nonpregnant cows were immediately eliminated from the herd and the pregnant cows were isolated and eliminated after parturiation. The calves with colostral antibodies were kept in the herd. There were no seropositive animals between the 6 to 12 months old calves and the heifers. The animals were monitored serologically every 3 months for 21 months and afterwords in 2 semestral intervals. The seropositive nonpregnant, pregnant and the nursing cows were the source of BHV-1 infecction. In order to maintain a BHV-1 free herd, measurements of control had been taken, as the utilization of virus-free semen, quarantine when animals were introduced into the herd, and semestral serological surveys. Through these measurements the farm has been BHV-1 free for 18 months.

Serological response of guinea pigs to oily and aqueous inactivated vaccines containing a Brazilian isolate of the Bovine Viral Diarrhea Virus (BVDV)

Bovine Viral Diarrhea Virus (BVDV) is widespread in cattle in Brazil and research shows its large antigenic variability. Available vaccines are produced with virus strains isolated in other countries and may not be effective. In this study, inactivated vaccines containing the Brazilian BVDV-Ib IBSP11 isolate were developed and tested on 6 groups of 10 guinea pigs (Cavia porcellus). Animals in groups A and C received an aqueous vaccine (aluminum hydroxide); B and D groups received an oily vaccine (Montanide ISA50); Group E positive-control animals were given an imported commercial vaccine with BVDV-Ia Singer; Group F animals were sham vaccinated (negative control). Groups A, B and E received two doses, and Groups C and D, three, every 21 days. Twelve blood samples were taken, at 21-day intervals over 231 days, and evaluated for antibody titer through virus-neutralization (VN), using a homologous strain (IBSP11), and a heterologous strain (BVDV-Ia NADL). Most animals, 42 days following the first dose, seroconverted to both strains and, after the second dose, there was a significant increase of titers in all groups. The oily formulation induced greater response after the third administration. This increase was not observed with the aqueous vaccines, regardless of the virus used in the VN. Antibody decline was more rapid in animals that received aqueous vaccines. The results showed the importance of studying the influence of endemic strains of commercial vaccines, to improve the efficacy of BVD vaccination. Use of the endemic strain in vaccine formulation presented promising results, as well as the use of guinea pigs as a laboratory model.

A experiência brasileira na erradicação da febre aftosa e o emprego do sistema I-ELISA 3ABC/EITB para certificação sanitária de bovinos e bubalinos

Vaccination against foot-and-mouth disease is an effective tool for eradication and prevention of this disease. However, the presence of non-capsidal proteins (NCP) in the vaccine, produced during viral replication, has been the main problem, since their presence hamper the vigilance as its relies on the detection of antibodies against NCP to differentiate vaccinated from infected animals. Therefore, the Brazilian Ministry of Agriculture, Livestock and Supply published the Normative Instruction 50 (IN 50) that included the detection of antibodies against the NCP to evaluate the removal of these proteins. Considering the vaccine interference, this paper aimed to evaluate the frequency of reagents to NCP, analyzed by I-ELISA 3ABC/EITB system in the Laboratory of Bovine Viruses (LVB), Biological Institute (IB), Sao Paulo, SP, Brazil, from 2002 to 2012. Of the 34,705 cattle examined, it was observed that the proportion of reagents to NCP increased with age, indicating increased frequency of reagents in animals that received more vaccines, showing interference of proteins in response to vaccination. When compared before and after the publication of IN 50, there was decreased reactivity, with reduction in 2010, of nearly half compared to 2007, and even higher when compared with 2002 to 2006. This shows the effectiveness of the purification of the vaccine in response to the implementation of IN 50, although it remains some seroreactivity in cattle with multiple vaccinations. The I-ELISA 3ABC/EITB system proved to be a great tool to prevent the movement of possible carriers of the virus derived from vaccinated herds, provided that all sanitary and epidemiological context are taken in consideration.

Pesquisa de anticorpos contra o vírus da Estomatite Vesicular em bovinos de corte criados na região de Araçatuba, Estado de São Paulo, Brasil em 2000

No ano de 2000 foram pesquisadas 1099 amostras de soro de bovinos de corte, adultos, pertencentes a seis rebanhos situados nos municipios de Auriflama, Gastao Vidigal,, Magda, Santo Antonio do Aracangua, Sebastianopolis do Sul e Turiuba, regiao de Aracatuba. A tecnica empregada para a dosagem de anticorpos para o virus da Estomatite Vesicular foi a soroneutralizacao em microplacas, utilizando a cepa Indiana 1 Costa Rica/72 com titulo de 106,0 DICT50/50mL. As celulas utilizadas foram as da linhagem de Rim de Macaco Verde Africano (VERO-CCL 81), provenientes do American Type Cell Colection (ATCC). O titulo de anticorpos de cada amostra de soro, foi calculado como a reciproca da maior diluicao do soro expresso em log10, que protegeu 100,00% da monocamada celular. Foram consideradas positivas as amostras com titulo superior a 1,6 log10 (1/40). Das 1099 amostras de soro analisadas, 28 (2,50%) foram positivas ao virus da Estomatite Vesicular tipo Indiana 1, com titulos de anticorpos variando de 1,9 a 2,8. Com excecao do rebanho situado no municipio de Turiuba, nos outros cinco foram encontrados animais reagentes ao virus. Esta avaliacao mostrou que houve circulacao do virus da Estomatite Vesicular nestes rebanhos provocando resposta humoral. Sendo assim e necessario a realizacao de estudos para se determinar o sitio de permanencia do virus.

Prevalence and risk factors for bluetongue in the State of São Paulo, Brazil

Abstract Bluetongue (BT), caused by Bluetongue virus (BTV), is a disease that affects ruminants such as cattle, sheep, goats and deer. BTV is transmitted by female midges of the genus Culicoides. In Brazil, information on the prevalence of BTV in cattle is limited, so the objective of this work was to identify BTV serotypes in cattle. The State of São Paulo was divided into seven cattle‐producing regions, and in each of them, 300 cattle farms were randomly selected. One animal from each farm (out of a total of 1,598 farms) was selected and its sera tested by virus neutralization technique against BTV serotypes (1–24 and 26) for determining antibody titre. Moreover, for each sampled farm, an epidemiological questionnaire was submitted to verify the type of cattle production and the zootechnical and sanitary practices carried out, which could be associated with a higher risk of BTV infection. In this study, antibodies (percentage, [95% confidence interval]) were identified against 11 serotypes: BTV‐1 (22.15%, [15.72–27.92]), BTV‐2 (31.03%, [26.65–37.98]), BTV‐3 (18.96%, [12.42–24.90]), BTV‐4 (24.90% [19.41–29.12]), BTV‐9 (6.82%, [1.45–11.72]), BTV‐12 (7.50%, [2.82–12.51]), BTV‐17 (23.90%, [17.35–29.35]), BTV‐19 (10.20%, [4.62–5.56]), BTV‐21 (30.66%, [25.00–36.00]), BTV‐22 (12.14%, [5.91–18.55]), BTV‐26 (57.00%, [51.41–63.59]). In this study, for the first time in Brazil serological evidence of the presence of serotypes BTV‐2, BTV‐9, BTV‐21 and BTV‐26 is reported. The variable ‘new cattle entering herd’ was considered a risk factor for the occurrence of infection (OR = 2.183, 95% CI = 1.6–2.9).

Seroprevalence of viral agents in vaquejada horses

In order to investigate the prevalence of viral diseases in vaquejada horses, samples were collected from Quarter Horse and Paint Horse breeds (males and females, aged between 2 and 15 years) from rural and urban areas of Mossoro, Rio Grande do Norte, Brazil were collected. The number of samples collected and tested varied with the techniques used: 132 serum samples were tested for vesicular stomatitis (VS) and equine viral arteritis (EVA), 118 for western and eastern equine encephalomyelitis (WEE/EEE), 114 for equine herpesvirus 1 (EHV-1), and 61 for equine influenza (EI). The techniques used varied according to the disease studied: tests for VS (Ribeirao and Alagoas strains), EVA, EHV-1, and WEE/EEE were performed using the seroneutralization technique in Vero cell cultures, while the test for for EI type-2 (H3N8) was performed using the hemagglutination inhibition technique. The following results were obtained: 39 (29.55%) and 6 (4.54%) VS-positive horses (n=132) harbored the Alagoas and Ribeirao strains, respectively; no animal tested positive for EVA (n=132); 11.01% (n=13) and 0.84% (n=1) animals tested positive for EEE and WEE (total n=118), respectively; and 22 out of 114 (19.29%) horses tested positive for EHV-1. The history of respiratory and reproductive diseases was statistically significantly (p = 0.02) related to the presence of antibodies against EHV-1. This is the first report of the seroprevalence of viral diseases in vaquejada horses in Rio Grande do Norte.Heider Irinaldo Pereira Ferreira1, Cecília Calabuig2, Pedro Augusto Cordeiro Borges1, Ilanna Vanessa Pristo de Medeiros Oliveira1, Débora Alves de Carvalho Freire3, Eliana Monteforte Cassaro Villalobos4, Maria do Carmo Custódio de Souza Hunold Lara4, Edviges Maristela Pituco4, Adriana Hellmeister de Campos Nogueira Romaldini5, Elenice Maria Sequetin Cunha4, Eliana De Stefano6, João Marcelo Azevedo de Paula Antunes7* 1Veterinarians, MSc. Hospital Veterinário Jerônimo Dix-Huit Rosado Maia, Universidade Federal Rural do Semiárido – UFERSA, Mossoró, RN, Brasil 2Biologist, Ph.D. Departamento de Biociências, Universidade Federal Rural do Semiárido – UFERSA, Mossoró, RN, Brasil 3Biotechnologist, MSc. Departamento de Ciência Animais, Universidade Federal Rural do Semiárido – UFERSA, Mossoró, RN, Brasil 4Veterinarians, Ph.D. Instituto Biológico, São Paulo, SP, Brasil 5Veterinary, MSc. Instituto Biológico, São Paulo, SP, Brasil 6Biologist, MSc. Instituto Biológico, São Paulo, SP, Brasil 7Veterinary, Ph.D. Hospital Veterinário Jerônimo Dix-Huit Rosado Maia, Universidade Federal Rural do Semiárido – UFERSA, Mossoró, RN, Brasil

Bovine leukaemia virus genotypes 5 and 6 are circulating in cattle from the state of São Paulo, Brazil

PURPOSE Enzootic bovine leucosis (EBL) is a silent disease caused by a retrovirus [bovine leukaemia virus (BLV)]. BLV is classified into almost 10 genotypes that are distributed in several countries. The present research aimed to describe two BLV gp51 env sequences of strains detected in the state of São Paulo, Brazil and perform a phylogenetic analysis to compare them to other BLV gp51 env sequences of strains around the world. METHODOLOGY Two bovines from different herds were admitted to the Bovine and Small Ruminant Hospital, School of Veterinary Medicine and Animal Science, University of São Paulo, Brazil. In both, lymphosarcoma was detected and the presence of BLV was confirmed by nested PCR. The neighbour-joining algorithm distance method was used to genotype the BLV sequences by phylogenetic reconstruction, and the maximum likelihood method was used for the phylogenetic reconstruction. The phylogeny estimates were calculated by performing 1000 bootstrap replicates. RESULTS Analysis of the partial envelope glycoprotein (env) gene sequences from two isolates (25 and 31) revealed two different genotypes of BLV. Isolate 25 clustered with ten genotype 6 isolates from Brazil, Argentina, Thailand and Paraguay. On the other hand, isolate 31 clustered with two genotype 5 isolates (one was also from São Paulo and one was from Costa Rica). The detected genotypes corroborate the results of previous studies conducted in the state of São Paulo, Brazil. The prediction of amino acids showed substitutions, particularly between positions 136 and 150 in 11 out of 13 sequences analysed, including sequences from GenBank. CONCLUSION BLV is still important in Brazil and this research should be continued.