ANÁLISE ESPACIAL EM SAÚDE: UTILIZAÇÃO EM SERVIÇOS DE SAÚDE ANIMAL

O sucesso do Programa Nacional de Profilaxia da Raiva Urbana só foi possível graças à realização de vacinações gratuitas e anuais em cães e gatos, executadas na forma de campanhas, em curto espaço de tempo e utilizando postos de vacinação temporários. Nesse sentido, qualquer ferramenta que possa aprimorar e adequar estratégias de vacinação nas campanhas é importante, sendo o geoprocessamento uma delas. O objetivo deste trabalho é apresentar as atividades desenvolvidas pelo grupo de extensão universitária, coordenado pela disciplina Planejamento de Saúde Animal e Saúde Pública da FMVZ Unesp/Botucatu, no que tange à elaboração de mapas com a distribuição geográfica dos postos de vacinação, estabelecidos em anos anteriores, visando gerar ferramentas para dar suporte ao planejamento da campanha antirrábica urbana de 2013, na cidade de Lençóis Paulista/SP. Visitas a cada local onde iria ser instalado o posto de vacinação foram realizadas pelo grupo de extensão e pelo residente da referida Disciplina, com suporte da universidade (transporte e um funcionário da prefeitura, com exímio conhecimento da área física da cidade). O georreferenciamento foi estabelecido com GPS móvel (Garmin Etrex Venture®). Para a confecção dos mapas, utilizou-se o ArcGIS 10.1® da empresa Esri. Foram visitadas 21 localidades do município, capturadas as coordenadas e verificadas as prerrogativas de um local

A UTILIZAÇÃO DO GEOPROCESSAMENTO EM CAMPANHAS DE VACINAÇÃO CONTRA A RAIVA DE CÃES E GATOS

Aliar saúde humana, saúde animal e saúde ambiental em torno da estratégia Um mundo, uma saúde não é uma tarefa fácil. A análise espacial (ambiente) em saúde é um aspecto fundamental na epidemiologia. Dentre as várias técnicas utilizadas para a análise espacial dos serviços de saúde, pode-se citar o geoprocessamento, que envolve etapas de coleta, tratamento e exibição de informações indexadas geograficamente. O objetivo deste trabalho é demonstrar que o geoprocessamento pode ser uma ferramenta útil no planejamento, execução, avaliação e elaboração de relatórios de campanhas de vacinação animal. Os dados utilizados são provenientes da 43ª Campanha de vacinação contra a raiva de cães e gatos de Botucatu/SP. Na fase de planejamento e execução utilizaram-se os pontos georreferenciados pela prefeitura; nas fases de avaliação e elaboração do relatório, os pontos utilizados foram aqueles georreferenciados no transcorrer da campanha com GPS móvel (Garmin Etrex Venture®), relativos aos locais onde estavam instalados os postos de vacinação temporários. Para a confecção dos mapas, utilizou-se o ArcGIS 10.1® da empresa Esri. Durante o planejamento e execução, a utilização dos mapas pôde orientar a distribuição dos postos, como nos novos loteamentos, com base na área de abrangência de 500 metros. Além disso, pôde facilitar a determinação dos postos transitórios e dos permanentes, e também

A utilização do geoprocessamento em campanhas de vacinação contra a raiva de cães e gatos

Aliar saúde humana, saúde animal e saúde ambiental em torno da estratégia Um mundo, uma saúde não é uma tarefa fácil. A análise espacial (ambiente) em saúde é um aspecto fundamental na epidemiologia. Dentre as várias técnicas utilizadas para a análise espacial dos serviços de saúde, pode-se citar o geoprocessamento, que envolve etapas de coleta, tratamento e exibição de informações indexadas geograficamente. O objetivo deste trabalho é demonstrar que o geoprocessamento pode ser uma ferramenta útil no planejamento, execução, avaliação e elaboração de relatórios de campanhas de vacinação animal. Os dados utilizados são provenientes da 43ª Campanha de vacinação contra a raiva de cães e gatos de Botucatu/SP. Na fase de planejamento e execução utilizaram-se os pontos georreferenciados pela prefeitura; nas fases de avaliação e elaboração do relatório, os pontos utilizados foram aqueles georreferenciados no transcorrer da campanha com GPS móvel (Garmin Etrex Venture®), relativos aos locais onde estavam instalados os postos de vacinação temporários. Para a confecção dos mapas, utilizou-se o ArcGIS 10.1® da empresa Esri. Durante o planejamento e execução, a utilização dos mapas pôde orientar a distribuição dos postos, como nos novos loteamentos, com base na área de abrangência de 500 metros. Além disso, pôde facilitar a determinação dos postos transitórios e dos permanentes, e também

Análise espacial em saúde: utilização em serviços de saúde animal

O sucesso do Programa Nacional de Profilaxia da Raiva Urbana só foi possível graças à realização de vacinações gratuitas e anuais em cães e gatos, executadas na forma de campanhas, em curto espaço de tempo e utilizando postos de vacinação temporários. Nesse sentido, qualquer ferramenta que possa aprimorar e adequar estratégias de vacinação nas campanhas é importante, sendo o geoprocessamento uma delas. O objetivo deste trabalho é apresentar as atividades desenvolvidas pelo grupo de extensão universitária, coordenado pela disciplina Planejamento de Saúde Animal e Saúde Pública da FMVZ Unesp/Botucatu, no que tange à elaboração de mapas com a distribuição geográfica dos postos de vacinação, estabelecidos em anos anteriores, visando gerar ferramentas para dar suporte ao planejamento da campanha antirrábica urbana de 2013, na cidade de Lençóis Paulista/SP. Visitas a cada local onde iria ser instalado o posto de vacinação foram realizadas pelo grupo de extensão e pelo residente da referida Disciplina, com suporte da universidade (transporte e um funcionário da prefeitura, com exímio conhecimento da área física da cidade). O georreferenciamento foi estabelecido com GPS móvel (Garmin Etrex Venture®). Para a confecção dos mapas, utilizou-se o ArcGIS 10.1® da empresa Esri. Foram visitadas 21 localidades do município, capturadas as coordenadas e verificadas as prerrogativas de um local

Validation of rapid and simple LC-MS/MS method for determination of voriconazole in rat plasma.

A rapid, simple and sensitive LC-MS/MS analytical method was developed and validated for the determination of voriconazole (VRC) in rat plasma, using ketoconazole as internal standard (IS). Analysis was performed on a Shimadzu HPLC system using a Shimadzu C18 column and isocratic elution with acetonitrile-water-formic acid (60:40:0.05, v/v/v), at a flow of 1.0 mL/min (split ratio 1:5), and a mass spectrometer Micromass, equipped with a double quadrupole and an electrospray ionization interface, operated in a positive mode. Plasma samples were deproteinized with methanol (1:2) and 30 microL of the supernatant was injected into the system. The retention times of VRC and IS were approximately 3.3 and 2.7 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 50-2500 ng/mL with determination coefficient >0.98. The lower limit of quantification was 50 ng/mL. The accuracy of the method was within 5%. Intra- and inter-day relative standard deviations were less or equal to 12.5 and 7.7%, respectively. The applicability of the LC-MS-MS method for pharmacokinetic studies was tested using plasma samples obtained after intravenous administration of VRC to male Wistar rats. The reported method provided the necessary sensitivity, linearity, precision, accuracy, and specificity to allow the determination of VRC in pre-clinical pharmacokinetic studies.

Molecular epidemiology of foot-and-mouth disease virus types A and O isolated in Argentina during the 2000-2002 epizootic.

During 2000-2002 a foot-and-mouth disease (FMD) epizootic affected Argentina and spread across the country resulting in more than 2500 outbreaks. In order to study the evolution of the FMD viruses (FMDV) and help with disease control measures, a genetic characterization and phylogenetic analysis was performed of 43 field isolates representative of the epizootic. The nucleotide sequence of the VP1-coding region was determined for the viruses and used in this study. Two serotype A lineages, A/Arg/00 and A/Arg/01 (1000/1000 bootstrap value) and two different serotype O/Arg/00 lineages (848/1000 bootstrap value) were identified. Phylogenetic analysis showed that viruses A/Arg/01 and O/Arg/00 could be related with former South American isolates, while the origin of A Argentina 2000 viruses remains unclear. Comparison of the amino acid sequences with vaccine reference strains revealed differences at critical antigenic sites for emergent strains A/Arg/00 and A/Arg/01, leading to a change in the current vaccine formulation.

Development and validation of a LC-MS/MS method with electrospray ionization for determination of LASSBio-579 in rat plasma.

A simple and sensitive LC-MS/MS analytical method was developed and validated for the determination of LASSBio-579 in plasma rat, using fluconazole as internal standard. Analyses were performed on a Shimadzu HPLC system using a Shimadzu C18 column and isocratic elution with acetonitrile-water (80:20, v/v), containing 0.4mM ammonium hydroxide and 0.2 mM acetic acid at a flow rate of 1.0 ml/min (split ratio 1:5). A Micromass triple quadrupole mass spectrometer, equipped with an electrospray ionization interface, operated in the positive mode. Plasma samples were deproteinized with acetonitrile (1:2) and 50 microl of the supernatant were injected into the system. The retention times of LASSBio-579 and IS were approximately 4.7 and 2.4 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 30-2000 ng/ml with determination coefficient >0.98. The lower limit of quantification was 30 ng/ml. The accuracy of method was within 15%. Intra- and inter-day relative standard deviations were less or equal to 13.5% and 6.4%, respectively. The applicability of the LC-MS/MS method for pharmacokinetic studies was tested using plasma samples obtained after intraperitoneal administration of LASSBio-579 to male Wistar rats. No interference from endogenous substances was observed, showing the specificity of the method developed. The reported method can provide the necessary sensitivity, linearity, precision, accuracy, and specificity to allow the determination of LASSBio-579 in pre-clinical pharmacokinetic studies.

Full length nucleotide sequence of foot-and-mouth disease virus strain O1 Campos/Bra/58. Brief report.

The complete nucleotide sequence of foot-and-mouth disease virus (FMDV) South American strain O(1) Campos/Bra/58 was determined. The 8,168 Kb sequence and the deduced amino acid sequence were compared to published FMDV sequences. They showed the highest sequence homology with the O(1) Kaufbeuren/FRG/66 strain, but closer evolutionary relatedness to the Argentinean strains.

BHV-1 DNA vaccination: effect of the adjuvant RN-205 on the modulation of the immune response in mice.

It is well documented that adjuvants improve the immune response generated by traditional viral vaccines, but less is known about the effects of adjuvants on the immune response elicited by DNA vaccines. In this study, we have investigated the use of RN-205 (immunomodulator containing a membrane rich in lipopolysaccharide from gram-negative bacteria) as an adjuvant and analyzed the humoral and cellular specific immune responses elicited by DNA vaccines based on the bovine herpesvirus-1 (BHV-1) glycoprotein D (gD). The comparison of the antibody response induced in mice by a mixture of the three different versions of DNA gD (membrane-anchored, secreted and cytosolic) formulated with or without RN-205 showed that the immunomodulator did not affect the total specific humoral response. The cellular immune response induced in mice immunized with vaccines plus RN-205 was higher than that obtained in mice vaccinated without RN-205, not only in the indexes of proliferation tests but in the number of IL-4 and gammaIFN secreting cells. When total spleen cells were marked with specific monoclonal antibodies against surface markers, a significant increase in the macrophage population of all the groups receiving RN-205 was observed. CD8 and CD4 positive cells were also increased but to a lesser extent. Our results indicate that the incorporation of RN-205 into DNA vaccines induces an increase of the cellular specific immune response in mice.

Phylogenetic analysis of foot-and-mouth disease viruses isolated in Argentina.

We have analysed complete or partial VPI sequences of 31 foot-and-mouth disease (FMD) viruses belonging to serotypes A, O and C to determine the genetic relatedness of field strains of FMD virus (FMDV) that have circulated in Argentina between 1961 and 1994. Phylogenetic analysis, which also included 15 previously published Argentinean sequences and six reference strains, revealed that (i) FMD type A strains showed the highest genetic heterogeneity and could be divided into five lineages with a sequence divergence of 0.9-18.5% between strains (ii) most of the FMD type O viruses grouped in two clusters (within cluster sequence divergence ranging from 0.2% to 6.0%) circulating in Argentina since the early 1960s, and (iii) FMD type C viruses were grouped in two clusters with a 13.4% nucleotide sequence divergence between each cluster. The availability of sequence data for many more field isolates from the region will enable us to understand the genetic relationships between FMDV strains and to rapidly trace the source of an FMD outbreak for epidemiological surveillance.

Biological and molecular characterization of a pigeon paramyxovirus type-1 isolate found in Argentina.

In this report, we describe the biological and molecular characterization of a paramyxovirus type-1 (PPMV-1) isolate found in wild pigeons in an urban habitat in Buenos Aires, Argentina. Of the nine pigeons captured, three were moribund, and the other six showed diarrhea, ataxia, tremor, torticolis, and wing paralysis. The intracerebral pathogenicity index was 1.29, and the amino acid (aa) sequence at the fusion protein cleavage site was 112GRQ KRF117. These characteristics correspond to a virulent Newcastle disease virus isolate. Nevertheless, it was not possible to reproduce the disease in chickens experimentally although the chickens exhibited seroconversion after inoculation. On the other hand, pigeons inoculated with the isolate became sick. These results provide further evidence about the unusual pathogenicity of PPMV-1 for chickens and show once more the need for more biological determinations in these cases to arrive at a final conclusion.

[Molecular characterization of aphthous fever virus isolated during the years 1993-1994 in Argentina]. / Caracterización molecular de cepas del virus de la fiebre aftosa aisladas durante los años 1993-1994 en la Argentina.

Nucleotide sequence and phylogenetic analysis of the VP1 structural protein have been used extensively as diagnostic and epidemiological tools for foot and mouth disease virus (FMDV). In this report we have applied this methodology to the analysis of the VP1 coding sequence from FMDV strains isolated in Argentina during 1993-1994. The results demonstrated that the field isolates were related to the vaccine strains used at that time. However the involvement of the vaccine virus appeared to be different for outbreaks caused by FMD viruses type O or C. These data provide a database essential for determining the origin of new epizootics.

[Expression and characterization of 2 bovine viral diarrhea virus structural proteins (]BVDV)]. / Expresión y caracterización de dos proteínas estructurales del virus de la diarrea viral bovina (VDVB).

The BVDV glycoproteins gp48 and gp53 were expressed in the baculovirus eukaryotic system. Both recombinant proteins were recognized in western blot analysis by monoclonal antibodies and polyclonal serum. Immunofluorescent test demonstrated that gp53 was localized on the cell surface whereas gp48 was in the cytoplasm. The expressed proteins were extracted by non-denaturing detergent treatment. Rabbit antiserum raised against gp53 recombinant protein efficiently neutralized the virus. These results demonstrate that the recombinant proteins have immunological properties similar to those of the native viral protein and that they can be useful as diagnostic reagents.

Caracterización molecular de cepas del virus de la fiebre aftosa aisladas durante los años 1993-1994 en la Argentina / Molecular characterization of aphthous fever virus isolated during the years 1993-1994 in Argentina

Nucleotide sequence and phylogenetic analysis of the VP1 structural protein have been used extensively as diagnostic and epidemiological tools for foot and mouth disease virus (FMDV). In this report we have applied this methodology to the analysis of the VP1 coding sequence from FMDV strains isolated in Argentina during 1993-1994. The results demonstrated that the field isolates were related to the vaccine strains used at that time. However the involvement of the vaccine virus appeared to be different for outbreaks caused by FMD viruses type O or C. These data provide a database essential for determining the origin of new epizootics.

Caracterización molecular de cepas del virus de la fiebre aftosa aisladas durante los años 1993-1994 en la Argentina / Molecular characterization of aphthous fever virus isolated during the years 1993-1994 in Argentina

Nucleotide sequence and phylogenetic analysis of the VP1 structural protein have been used extensively as diagnostic and epidemiological tools for foot and mouth disease virus (FMDV). In this report we have applied this methodology to the analysis of the VP1 coding sequence from FMDV strains isolated in Argentina during 1993-1994. The results demonstrated that the field isolates were related to the vaccine strains used at that time. However the involvement of the vaccine virus appeared to be different for outbreaks caused by FMD viruses type O or C. These data provide a database essential for determining the origin of new epizootics.(AU)

[Smoking time evolution among medical students, 1986, 1991, 1996]. / Evolução temporal do tabagismo em estudantes de medicina, 1986, 1991, 1996.

OBJECTIVE: To evaluate trends in smoking habits among medical students in the last ten years. METHODS: In 1996, a cross-sectional survey of smoking habits was carried out among students in the first to the fifth year of medical school. Four hundred and nine students answered the questionnaire. A regular smoker was defined as someone who smokes one or more cigarettes a day at least for one month; former smokers were the ones who used to smoke in the past but not at the moment. Similar research was conducted in 1986 and 1991. RESULTS/CONCLUSIONS: Smoking prevalence was 11% compared with 14% in 1991 and 21% in 1986. Although there was a significant reduction of smoking in the last three years, the actual decrease from 1991 to 1996 was less than that observed from 1986 to 1991. In 1961, the prevalence of smoking increased in the last years of medical school. There were no significant differences regarding gender. Most of the students were in favor of not allowing smoking in both the school areas and hospital wards. They also mentioned in their questionnaire that smoking was poorly approached during medical school.

Identification of optimal regions for phylogenetic studies on VP1 gene of foot-and-mouth disease virus: analysis of types A and O Argentinean viruses.

An analysis of the informative content of sequence stretches on the foot-and-mouth disease virus (FMDV) VPI gene was applied to two important viral serotypes: A and O. Several sequence regions were identified to allow the reconstruction of phylogenetic trees equivalent to those derived from the whole VPI gene. The optimal informative regions for sequence windows of 150 to 250 nt were predicted between positions 250 and 550 of the gene. The sequences spanning the 250 nt of the 3' end (positions 400 to 650), extensively used for FMDV phylogenetic analyses, showed a lower informative content. In spite of this, the use of sequences from this region allowed the derivation of phylogenetic trees for type A and type O FMDVs which showed topologies similar to those previously reported for the whole VP1 gene. When the sequences determined for viruses isolated in Argentina, between 1990 and 1993, were included in these analyses, the results obtained revealed features of the circulation of type A and type O viruses in the field, in the months that preceded the eradication of the disease in this country. Type A viruses were closely related to an Argentinean vaccine strain, and defined an independent cluster within this serotype. Among the type O viruses analysed, two groups were distinguished; one was closely related to the South American vaccine strains, while the other was grouped with viruses of the O3 subtype. In addition, a detailed phylogeny for type A FMDV is presented.

Expresión y caracterización de dos proteínas estructurales del virus de la diarrea viral bovina (VDVB) / Expression and characterization of 2 bovine viral diarrhea virus structural proteins (]BVDV)

The BVDV glycoproteins gp48 and gp53 were expressed in the baculovirus eukaryotic system. Both recombinant proteins were recognized in western blot analysis by monoclonal antibodies and polyclonal serum. Immunofluorescent test demonstrated that gp53 was localized on the cell surface whereas gp48 was in the cytoplasm. The expressed proteins were extracted by non-denaturing detergent treatment. Rabbit antiserum raised against gp53 recombinant protein efficiently neutralized the virus. These results demonstrate that the recombinant proteins have immunological properties similar to those of the native viral protein and that they can be useful as diagnostic reagents.