Com o objetivo de otimizar a avaliação de eficácia de vacinas contra o sarampo, discriminando-se imunização primária de secundária com uma única amostra pós-vacinal, o teste de avidez de anticorpos lgG anti-sarampo foi padronizado e avaliado no presente estudo. Aplicando-se a técnica de ELISA e a uréia 7M durante 10 minutos como agente dissociante da ligação antígeno-anticorpo, determinou-se o cut-off do baixo índice de avidez (BIA) para definição de primo-vacinação como 29 por cento e o tempo adequado de coleta da amostra pós-vacinal como 1O semanas. Observou-se BIA em todas as 164 amostras de soro colhidas até 1O semanas após primo- vacinação aos 9 meses com as vacinas Biken-CAM 70 e Edmonston-Zagreb. Após primo-vacinação aos 9 meses com a vacina Schwarz, BIA foi detectado em 233 das 242 (96,3 por cento) amostras colhidas em papel de filtro. No grupo de 41 crianças vacinadas que apresentavam história prévia de vacinação e/ou anticorpos na amostra pré-vacinal (reinfecção), 39/41 (95,1 por cento) das amostras apresentaram altos índices de avidez. Falhas vacinais primária e secundária foram detectadas, respectivamente, em 1/5 e 4/5 amostras de crianças que tinham história de vacinação, mas ausência de anticorpos na amostra pré-vacinal. Nenhuma das 90 amostras de crianças vacinadas no passado com 2 doses da vacina Biken-CAM70 (aos 6 e 11 meses) e nenhum dos 42 soros de cordão umbilical, apresentou BIA. Conclui-se que a determinação da avidez de anticorpos lgG contra o vírus do sarampo, pela técnica utilizada no presente estudo é altamente eficaz na discriminação entre primo-vacinação e reinfecção ou infecção passada, podendo ser utilizada na avaliação de eficácia de vacinas contra o sarampo empregando-se apenas uma amostra pós-vacinal. / In an attempt to improve methods for evaluation of measles vaccine efficacy, by discriminating primary from secondary immunization with only one post- vaccination sample, a measles lgG antibody avidity assay was standardized and evaluated. Using ELISA and treatment with 7M urea for 1O minutes to dissociate antigen- antibody bounds, the cut-off of low avidity indices (LAI) after primary measles vaccination was defined as 29 per cent and 1O weeks as the term for collection of post- vaccination samples. Low avidity indices were observed in all 164 samples collected up to 1 O weeks after primary vaccination at 9 months with Biken-CAM70 and Edmonston-Zagreb vaccines. After vaccination with Schwarz vaccine, LAI were detected in 233 of 242 (96.3 per cent) filter paper blood samples. In the group of 41 vaccinated children who had a history of previous vaccination and/or measles antibody in pre-vaccination samples (reinfection), all but two (95,1 per cent) presented high avidity indices. Primary and secondary measles vaccine failures were detected, respectively, in 1/4 and 4/5 children who had a history of previous vaccination but no measles antibody was detected in pre-vaccination samples. None of 90 children vaccinated in the past with 2 doses of measles vaccine (at 6 and 11 months) and none of 42 cord sera had LAI. We conclude that the measles antibody avidity antibody assay presented in this study is highly efficient in discriminating primary vaccination from reinfection or past infection and can be applied in the evaluation of measles vaccine efficacy, using only one post-vaccination sample.

Com o objetivo de otimizar a avaliação de eficácia de vacinas contra o sarampo, discriminando-se imunização primária de secundária com uma única amostra pós-vacinal, o teste de avidez de anticorpos lgG anti-sarampo foi padronizado e avaliado no presente estudo. Aplicando-se a técnica de ELISA e a uréia 7M durante 10 minutos como agente dissociante da ligação antígeno-anticorpo, determinou-se o cut-off do baixo índice de avidez (BIA) para definição de primo-vacinação como 29 por cento e o tempo adequado de coleta da amostra pós-vacinal como 1O semanas. Observou-se BIA em todas as 164 amostras de soro colhidas até 1O semanas após primo- vacinação aos 9 meses com as vacinas Biken-CAM 70 e Edmonston-Zagreb. Após primo-vacinação aos 9 meses com a vacina Schwarz, BIA foi detectado em 233 das 242 (96,3 por cento) amostras colhidas em papel de filtro. No grupo de 41 crianças vacinadas que apresentavam história prévia de vacinação e/ou anticorpos na amostra pré-vacinal (reinfecção), 39/41 (95,1 por cento) das amostras apresentaram altos índices de avidez. Falhas vacinais primária e secundária foram detectadas, respectivamente, em 1/5 e 4/5 amostras de crianças que tinham história de vacinação, mas ausência de anticorpos na amostra pré-vacinal. Nenhuma das 90 amostras de crianças vacinadas no passado com 2 doses da vacina Biken-CAM70 (aos 6 e 11 meses) e nenhum dos 42 soros de cordão umbilical, apresentou BIA. Conclui-se que a determinação da avidez de anticorpos lgG contra o vírus do sarampo, pela técnica utilizada no presente estudo é altamente eficaz na discriminação entre primo-vacinação e reinfecção ou infecção passada, podendo ser utilizada na avaliação de eficácia de vacinas contra o sarampo empregando-se apenas uma amostra pós-vacinal. / In an attempt to improve methods for evaluation of measles vaccine efficacy, by discriminating primary from secondary immunization with only one post- vaccination sample, a measles lgG antibody avidity assay was standardized and evaluated. Using ELISA and treatment with 7M urea for 1O minutes to dissociate antigen- antibody bounds, the cut-off of low avidity indices (LAI) after primary measles vaccination was defined as 29 per cent and 1O weeks as the term for collection of post- vaccination samples. Low avidity indices were observed in all 164 samples collected up to 1 O weeks after primary vaccination at 9 months with Biken-CAM70 and Edmonston-Zagreb vaccines. After vaccination with Schwarz vaccine, LAI were detected in 233 of 242 (96.3 per cent) filter paper blood samples. In the group of 41 vaccinated children who had a history of previous vaccination and/or measles antibody in pre-vaccination samples (reinfection), all but two (95,1 per cent) presented high avidity indices. Primary and secondary measles vaccine failures were detected, respectively, in 1/4 and 4/5 children who had a history of previous vaccination but no measles antibody was detected in pre-vaccination samples. None of 90 children vaccinated in the past with 2 doses of measles vaccine (at 6 and 11 months) and none of 42 cord sera had LAI. We conclude that the measles antibody avidity antibody assay presented in this study is highly efficient in discriminating primary vaccination from reinfection or past infection and can be applied in the evaluation of measles vaccine efficacy, using only one post-vaccination sample.

The mouse parvovirus designated MPV can infect laboratory mice and affect the humoral and cellular immune response of infected mice, reducing their value for biomedical and medical research. The development and maintenance of MPV-free mouse colonies for biomedical research is therefore essential and requires routine monitoring of the infection status of mice, using serological surveillance procedures. Recent experience in the Animal Resources Centre (ARC), a major supplier of mice to the medical research community in Australia, was that MPV infection was present but was not detectable with the serological tests that were then in routine use. This thesis reports the development of a polymerase chain reaction (PCR) assay for the detection of the MPV in the ARC mouse colonies, the genetic characteristics of the strain of MPV detected, the development of a recombinant virus protein that provided a suitable antigen for enzyme-linked immunosorbent assay (ELISA) and a Western immunoblot (WIB) assay for the detection of MPV antibodies, and use of these various assays to determine aspects of the epidemiology and pathogenicity of the infection that were critical to the eradication of virus infection and future immunological surveillance to ensure the absence of infection. The recombinant protein produced as an antigen was a biotinylated fusion protein, a truncated capsid protein of the strain of MPV detected in the ARC, and was produced using the PinpointTM vector and with expression in Escherichia coli. The protein was produced as an insoluble intracellular product within inclusion bodies and was solubilised using urea and purified. The purified protein was utilised as an antigen for ELISA and the WIB assays to detect virus antibody in infected mice. The outbreak of MPV infection in the ARC was used as an unique opportunity for assessment of the seroprevalence of MPV-1 infection in a large laboratory mouse colony and to utilise this data to determine the sampling size needed to reliably detect MPV-1 infection within such large laboratory mouse colonies. An overall seroprevalence of 16.5% was detected using the developed serological tests, but considerable variation in prevalence was detected in different mouse strains. The response to MPV infection of 4 different but common strains of mice was determined as a basis for developing appropriate surveillance procedures and the selection of appropriate sentinel animals. The effect of infection of these strains at different ages was also investigated. Virus replication was detected in tissues of all the mice strains infected (outbred ARC(s) and inbred C57BL/6JArc, BALB/c and BALB/c-Foxn1nu/Arc) as juveniles and adults, with the exception of C57BL/6JArc inoculated as adults. However, while seroconversion in mice inoculated as juveniles and adults was detected in ARC(s) and C57BL/6JArc mice, it was not detected in BALB/c mice. The high rate of seroconversion to MPV, the early and prolonged development of an immune response, and the lack of age differences in their susceptibility indicated that ARC(s) mice would provide reliable sentinels for the detection of MPV. The genomic nucleotide sequence of the ARC strain, excluding the terminal palindromic regions and the predicted amino acid sequences of the non-structural and structural proteins was determined. This strain was very similar (98-99% nucleotide identity) to the previously described MPV strains MPV-1a, MPV-1b and MPV -1c. The similarity suggested there were unlikely to be significant antigenic differences in the proteins of the ARC strain and those strains of MPV reported previously.

Žmogaus plaukas – neatsiejama žmogaus organizmo dalis. Lietuvos kriminalistikos moksle plaukas priskiriamas biologinių objektų grupei, o tiriant įvairius nusikaltimus, neretai tampa puikiu informacijos šaltiniu. Baigiamajame darbe pateikiama trumpa kriminalistinio žmogaus plaukų tyrimo istorinė raida Lietuvos prieškario ir pokario laikotarpiu. Trumpai yra aprašoma žmogaus plauko struktūra. Pateikiami trys pagrindiniai kriminalistiniai plaukų tyrimo metodai: morfologinis, serologinis ir DNR analizės. Aptariamos šių metodų atlikimo galimybės Lietuvoje.Palyginimui pateikiamos naujausios žmogaus plaukų tyrimo metodikos užsienio šalyse (Anglijoje, Vokietijoje, Švedijoje) – žmogaus plauko kutikulės išliejos metodas; botaninės kilmės aplinkos dalelių (žiedadulkių ir sporų) žmogaus plaukų nuoplovose metodas; katageninių ir telogeninių plaukų DNR tyrimai, pasitelkus „LCN sistemą“; geno (MC1R), apsprendžiančio žmogaus plaukų spalvą ir odos tipą, mutacijų tyrimas; narkotinių medžiagų žmogaus galvos ir gaktos plaukuose nustatymo tyrimai. Analizuojama Lietuvos ekspertinių įstaigų 2004-2006 metų veikla, susijusi su kriminalistiniu žmogaus plaukų tyrimu. Aptariami ekspertų-specialistų anketinės apklausos rezultatai. Pateikiama išsami ekspertizės aktų analizė, statistiniai duomenys apie nusikaltimus, kurių aplinkybėms aiškinti buvo tiriami žmogaus plaukai, įrodoma žmogaus plaukų, kaip tyrimo objekto kriminalistikoje, reikšmė, tiriant smurtinius ir... [toliau žr. visą tekstą] / Human hair – a part of human body. Lithuanian Forensic Science it presents like the group of biological evidences. Due investigation of the variuos commited crimes, human hair becomes a wide source of important and useful informacion. Noted a short Lithuanian prewar and postwar history of forensic human hair exploration. Shortly described a structure of human hair. There were mentioned three, the main ways of forensic investigation of human hair in Lithuania: morphological, serological exploration and DNA analysis. The procedures and the opportunities of these methods applied in Lithuania, were discussed as well. Described the modern procedures of human hair investigation: a scale cast obtained from shaft of human hair; the exploration of polynomorphs (pollens and spores) in human hair; the DNA analysis of human catagen and telogen hair by Low Copy Number (LCN) method; a single gene (MC1R) extraction for identification of human hair colors from degraded DNR; the human hair analysis for drugs detection in it. Submited a comprehensive assay of activity in Lithuanian Forensic Institutes, which is dealing with forensic human hair investigation. There was offered an importance of human hair exploration in forensic trial. Equally, were presented an inquiry results of Lithuanian forensic experts. Were discussed a generalization of forensic experts reports, shown a statistic of commited crimes, where the human hair was found and took like... [to full text]

Ovarian cancer has very high mortality because it is hard to diagnosis in early stages. Many ovarian cancer biomarkers (such as HE4, CA 125) are available and had been suggested as potential tools for early cancer detection. However, early cancer detection using serological markers will only become widely used if a new generation of sensors that can be handled in a clinical setting can be developed. A detection technology that is promising for miniaturization and integration in biomedical sensing devices is based on the phenomenon of the extraordinary light transmission (EOT) through arrays of nanoholes on metal films. EOT is an increase in light transmission observed at certain wavelengths that satisfy the surface plasmon resonance (SPR) condition of the nanostructure. The position of this resonance is affected by surface adsorption phenomenon, which is the basis for the biosensor. In this dissertation, the detection of the HE4 biomarker was demonstrated using EOT. The EOT-based detection was compared to two state-of-the-art analytical methods (ELISA and commercial SPR). Based on our experiments, it was found that ELISA has lowest detection limit, around 0.5 ng/mL for that particular protein (HE4). The detection limits for the commercial SPR, around 0.13 μg/mL was comparable to the nanohole-based detection limit, around 1.76 μg/mL. In contrast to ELISA, the SPR-based methods were label free, more time efficient, and more environmental friendly. An extra advantage of the nanohole scheme was that multiple samples could be analyzed simultaneously and in real time. Adsorption kinetic experiments were also performed to evaluate the rate constants of the HE4 binding to a surface coated with anti-HE4. The adsorption equilibrium constant for the HE4 – anti-HE4 system was determined to be (4.3 ± 2.1) x 107 M-1. / Graduate / 0487

In the rural areas of Kwa Zulu Natal small scale farming is the main agricultural activity, which is often carried out in addition to other employment taken on in another location by at least one member of the household. Although Veterinary services (VS) was aware of several diseases occurring in this area and has implemented a dipping strategy for tick borne diseases as well as a regional annual vaccination campaign for Rabies, Anthrax and Black quarter, uncertainty remained about the relative importance of other diseases such as brucellosis, leptospirosis and enzootic bovine leucosis. Further it was of interest to investigate the serological resistance of cattle against the tick borne diseases babesiosis and anaplasmosis. In addition to this veterinary services wanted to increase their understanding of the perceptions and practices of local cattle owners that were relevant for the context of disease control. A serological survey of cattle was carried out between March 2001 and December 2003 to determine the prevalence of the above-mentioned diseases. The survey was designed as a two-stage survey, considering the diptank as the primary sampling unit. The conduction of the questionnaire survey was based on a convenience sample and took place during the dipping procedure. The apparent prevalence at district level was adjusted for clustering, and diagnostic test sensitivity and specificity and displayed in maps. The mean true prevalence of brucellosis varied from zero to 15.8 percent in the north eastern region with the large majority of the districts being disease free. Enzootic Bovine Leukosis (EBL) was widely present in the province at generally low prevalences, except in the central region where the highest prevalence at district level was recorded to be 70 percent. Leptospirosis also occurred frequently with the highest prevalence noted being 62 percent at district level. The southern regions showed a higher leptospirosis prevalence than other areas of the province, while in some of the northern and western districts a lower prevalence is noted. The encountered servovars were pomona, which occurred most frequently, tarrasovi, bratislava, hardjo, canicola and icterohaemorrhagica. While Babesia bovis and anaplasmosis occurred at a high prevalence throughout the province, B. bigemina was found to be much less established and is absent from many of the northern districts. Most prevalences calculated at district level do show large confidence intervals due to uncertainty that arose from the sampling frame and should be interpreted with care. / Dissertation (MSc (Production Animal Studies))--University of Pretoria, 2007. / Production Animal Studies / MSc / unrestricted

The investigation presented here was accomplished using agglutination and agar gel immunodiffusion techniques to compare Azotobacter agilis s 3at, A. chroococcum Italy AC 16, A. macrocytogenes St. M and A. vinelandii ATCC 12837. It was found that the agglutination titers differed sufficiently to allow partial identification of the four species. The homologous and heterologous systems studied by agar gel immunodiffusion tests showed that each of the four Azoto bacter species differed sufficiently in their soluble antigens to give distinct, identifiable patterns of antigen-antibody reactions on Ouchterlony agar plates. These studies also showed several antigens common to the four species tested and the resultant antigen-antibody cross reactions. The results of these investigations suggest that this approach opens a new avenue for the identification of the organisms of genus Azotobacter and perhaps, by extension, the family Azotobacteraceae.

Uma das maiores preocupações relacionadas a segurança transfusional é a possibilidade de transmissão de doenças infecciosas atraves de sangue transfundido, entre as quais se encontram as hepatites B e C, o HIV e a sífilis. Este trabalho teve como objetivo estudar doadores que passaram pelo processo de seleção pré-doação no Hemocentro de Ribeirão Preto e apresentaram resultados de testes sorológicos positivos para essas doenças/infecções.Estudou-se a frequencia dos resultados positivos entre os individuos que doaram sangue no hemocentro de Ribeirão Preto ou em seu posto de Coleta Central,de 1o.de julho de 2005 a 31 de julho de 2006. Os indivíduos com resultados sorológicos positivos confirmados em um segundo teste foram caracterizados segundo algumas variáveis demográficas e socioeconomicas, tendo sido identificados seus fatores de risco e as causas determinantes de sua não-detecção na triagem clínica (TC). Estudaram-se também o comportamento de doadores em relação a auto-exclusão confidencial (AEC) e os fatores determinantes desse comportamento. Participaram da pesquisa 106 doadores, predominantemente do sexo masculino e casados, com menos de 40 anos de idade e baixa escolaridade. Eram principalmente doadores de primeira vez, procedentes de Ribeirão Preto e da região de Ribeirão Preto e pertencentes aos estratos econômicos C e D. As frequencias de marcadores sorologicos positivos encontradas foram: 0,07% para o HBsAg; 0,03% para o anti-HIV; 0,13% para o VDRL; 0,21 para o anti-HCV. Cerca de 40% dos participantes assumiram ter omitido fatores de risco na TC. Os motivos mencionados foram: não se sentir a vontade para falar ou não achar relevante fazê-lo; confiar totalmente nos exames; ter como objetivo conhecer sua condição sorológica; encontrar problemas relacionados a entrevista/triador; não confiar no sigilo das informações; sentir constrangimento diante de acompanhantes. Apenas 1,9% dos participantes utilizaram a AEC, e as justificativas alegadas foram: estar se sentindo bem e, por isso, não julgar necessário; não se considerar de risco;não achar que tivesse mentido na entrevista. Os achados indicam a necessidade de mudar a abordagem dos doadores na TC, de rever os procedimentos de captação de doadores e de reavaliar profundamente os procedimentos de orientação/conscientização dos candidatos a doação, procurando tornar esses procedimentos mais eficazes. / One of the main concerns related to the safety of transfusion is the possibility of diseases/infections transmitted through blood transfusion,amongst which we may include B and C hepatitis, HIV and syphilis.This paper aimed at studying the donors who have undergone the pre-donation selection at Blood Center of Ribeirao Preto and have presented positive serological results for theses diseases/infections.We have studied the frequency of positive results amongst the individuals who have donate blood at Blood Center of Ribeirao Preto and at the Central Collection Unit, from July 1st 2005 to July 31st 2006. The individuals who have had their positive serological results confirmed at a second test were classified according to demographic and socioeconomic variables, the risk factors present amongst them were identified, as well as the determinant causes for detection failure at the clinical screening. A study on the donors\' behavior towards the confidential self-exclusion process and the determinant factors that caused such behavior was also carried out. The research reached 106 donors, most of whom were males, married, under 40 and with low level of education. Most of them were first-time donors from the City of Ribeirão Preto and surrounding towns, belonging to the C and D economic classes. The frequencies of the serological markers were: 0,07% to HbsAg; 0,03% to anti-HIV; 0,13% to VDRL; 0,21 to anti-HCV. Around 40% of the participants affirmed having omitted risk factors at the clinical screening (CS). The reasons mentioned were the following: feeling unease to talk about it or not finding it relevant; relying completely on test results; having the aim of being aware of their real serological condition; issues related to the interview or screener; lack of trust in the confidentiality of the information he/she would give; feeling constrained in the presence of their companions. Only 1.9% of the participants used the confidential self-exclusion process, and the reasons given as justifications were: being feeling well and therefore not finding it necessary; not considering himself/herself a risk; not agreeing that he/she had lied during the interview. The findings indicate the necessity of changes as to the approach to donors at the CS, the need of a review of the procedures carried out to motivate people to become donors, and a deep revaluation of the procedures for orientation/awareness by candidates as to donation, aiming at turning them more effective.

Uma das maiores preocupações relacionadas a segurança transfusional é a possibilidade de transmissão de doenças infecciosas atraves de sangue transfundido, entre as quais se encontram as hepatites B e C, o HIV e a sífilis. Este trabalho teve como objetivo estudar doadores que passaram pelo processo de seleção pré-doação no Hemocentro de Ribeirão Preto e apresentaram resultados de testes sorológicos positivos para essas doenças/infecções.Estudou-se a frequencia dos resultados positivos entre os individuos que doaram sangue no hemocentro de Ribeirão Preto ou em seu posto de Coleta Central,de 1o.de julho de 2005 a 31 de julho de 2006. Os indivíduos com resultados sorológicos positivos confirmados em um segundo teste foram caracterizados segundo algumas variáveis demográficas e socioeconomicas, tendo sido identificados seus fatores de risco e as causas determinantes de sua não-detecção na triagem clínica (TC). Estudaram-se também o comportamento de doadores em relação a auto-exclusão confidencial (AEC) e os fatores determinantes desse comportamento. Participaram da pesquisa 106 doadores, predominantemente do sexo masculino e casados, com menos de 40 anos de idade e baixa escolaridade. Eram principalmente doadores de primeira vez, procedentes de Ribeirão Preto e da região de Ribeirão Preto e pertencentes aos estratos econômicos C e D. As frequencias de marcadores sorologicos positivos encontradas foram: 0,07% para o HBsAg; 0,03% para o anti-HIV; 0,13% para o VDRL; 0,21 para o anti-HCV. Cerca de 40% dos participantes assumiram ter omitido fatores de risco na TC. Os motivos mencionados foram: não se sentir a vontade para falar ou não achar relevante fazê-lo; confiar totalmente nos exames; ter como objetivo conhecer sua condição sorológica; encontrar problemas relacionados a entrevista/triador; não confiar no sigilo das informações; sentir constrangimento diante de acompanhantes. Apenas 1,9% dos participantes utilizaram a AEC, e as justificativas alegadas foram: estar se sentindo bem e, por isso, não julgar necessário; não se considerar de risco;não achar que tivesse mentido na entrevista. Os achados indicam a necessidade de mudar a abordagem dos doadores na TC, de rever os procedimentos de captação de doadores e de reavaliar profundamente os procedimentos de orientação/conscientização dos candidatos a doação, procurando tornar esses procedimentos mais eficazes. / One of the main concerns related to the safety of transfusion is the possibility of diseases/infections transmitted through blood transfusion,amongst which we may include B and C hepatitis, HIV and syphilis.This paper aimed at studying the donors who have undergone the pre-donation selection at Blood Center of Ribeirao Preto and have presented positive serological results for theses diseases/infections.We have studied the frequency of positive results amongst the individuals who have donate blood at Blood Center of Ribeirao Preto and at the Central Collection Unit, from July 1st 2005 to July 31st 2006. The individuals who have had their positive serological results confirmed at a second test were classified according to demographic and socioeconomic variables, the risk factors present amongst them were identified, as well as the determinant causes for detection failure at the clinical screening. A study on the donors\' behavior towards the confidential self-exclusion process and the determinant factors that caused such behavior was also carried out. The research reached 106 donors, most of whom were males, married, under 40 and with low level of education. Most of them were first-time donors from the City of Ribeirão Preto and surrounding towns, belonging to the C and D economic classes. The frequencies of the serological markers were: 0,07% to HbsAg; 0,03% to anti-HIV; 0,13% to VDRL; 0,21 to anti-HCV. Around 40% of the participants affirmed having omitted risk factors at the clinical screening (CS). The reasons mentioned were the following: feeling unease to talk about it or not finding it relevant; relying completely on test results; having the aim of being aware of their real serological condition; issues related to the interview or screener; lack of trust in the confidentiality of the information he/she would give; feeling constrained in the presence of their companions. Only 1.9% of the participants used the confidential self-exclusion process, and the reasons given as justifications were: being feeling well and therefore not finding it necessary; not considering himself/herself a risk; not agreeing that he/she had lied during the interview. The findings indicate the necessity of changes as to the approach to donors at the CS, the need of a review of the procedures carried out to motivate people to become donors, and a deep revaluation of the procedures for orientation/awareness by candidates as to donation, aiming at turning them more effective.

A ferramenta mais importante para o controle da brucelose em bubalinos e bovinos é a vacinação compulsória, utilizando-se a vacina viva atenuada contendo a estirpe B19 de Brucella abortus. A vacina B19 confere imunidade duradoura à infecção nos animais vacinados, e sua aplicação é restrita às fêmeas com idade entre 3 e 8 meses. A vacina B19 induz a produção de anticorpos que interferem no sorodiagnóstico da infecção, de forma que o diagnóstico sorológico de animais vacinados deve ser realizado apenas após os 24 meses de idade, idade na qual os anticorpos de origem vacinal já declinaram para níveis não detectáveis. O desempenho, a cinética humoral e o período de clearance da estirpe vacinal são bem conhecidos nos bovinos, contudo há poucas informações referentes a estes aspectos na espécie bubalina. O presente trabalho teve como objetivo avaliar a eliminação da estirpe B19 pelas fêmeas bubalinas vacinadas com a dose padrão contendo de 60 a 120 bilhões de UFC, assim como o perfil da resposta imune humoral induzido pela vacinação e a eficiência dos principais testes laboratoriais em diferenciar anticorpos vacinais dos anticorpos induzidos pela infecção natural. Foram utilizadas 44 fêmeas bubalinas provenientes de duas propriedades, negativas para brucelose, localizadas no interior do Estado de São Paulo, sendo 33 animais da propriedade A e 11 da propriedade B. Os animais foram vacinados e amostras de sangue, swab vaginal e urina foram coletas semanalmente, desde o dia em que a vacinação foi realizada (D0) até que os animais completassem 24 meses de idade. Os soros obtidos a partir das amostras de sangue foram utilizados na avaliação sorológica, empregando-se os seguintes testes: antígeno acidificado tamponado (AAT), 2-mercaptoetanol (2ME), fixação de complemento (FC) e polarização fluorescente (PF). As amostras de urina e swab vaginal foram submetidas ao cultivo microbiológico para isolamento de Brucella spp. e à reação em cadeia pela polimerase (PCR) para avaliar a eliminação da estirpe vacinal. Na PCR, as amostras foram amplificadas primeiramente utilizando-se primers direcionados à região interespaçadora do RNA ribossomal de Brucella spp. (PCR-ITS) para a detecção de qualquer componente do gênero. Amostras com resultados positivos pela PCR-ITS foram submetidas à PCR utilizando-se primers direcionados ao gene eri de Brucella spp, num ensaio específico para a detecção da estirpe B19 (PCR-B19). Foi calculada a especificidade de cada teste sorológico em detectar como negativos os animais vacinados. Todos os animais apresentaram-se reagentes aos sete dias pós-vacinação no teste de AAT, aos 14 dias no teste de 2ME, aos 126 no teste de FC. No teste de PF, o D28 foi o período em que o maior número de animais apresentaram resultados positivos. Dezesseis animais não soroconverteram em nenhum momento do experimento pelo teste de PF. A PF foi o teste mais eficiente na diferenciação de anticorpos vacinais, sendo que todos os animais apresentaram resultados negativos aos 151 e 232 dias pós-vacinais nas propriedades A e B, respectivamente. No teste de FC, 100% animais apresentaram resultados negativos no D552 (propriedade A) e no D331 (propriedade B). No D552 na propriedade A as especificidades foram de 92,31% para o teste de 2ME e 69,23% para o teste de AAT. A propriedade B apresentou nos dias D366 especificidades de 72,73% e 36,36% para os testes 2ME e AAT, respectivamente. Todos os animais foram negativos pelo cultivo microbiológico em todo o período de monitoramento. Na PCR foram verificadas 19 das 1144 amostras positivas (1,66%). Das 44 bezerras avaliadas, 15 (34,1%) apresentou resultado positivo em pelo menos uma colheita, em amostras de urina e/ou swab vaginal. De acordo com os resultados apresentados, pode-se concluir que a prova sorológica mais eficiente para a diferenciação dos anticorpos de origem vacinal dos da infecção natural é o teste de PF e que, a eliminação da estirpe vacinal pode ocorrer de forma intermitente, durante períodos prolongados nas fêmeas bubalinas vacinadas. / The most important tool for the control of brucellosis in buffalo and cattle is compulsory vaccination, using a live and attenuated vaccine containing the S19 strain of Brucella abortus. The S19 vaccine confers long-lasting immunity to infection in vaccinated animals and the application is restricted to females aged between 3 and 8 months. The S19 vaccine induces production of antibodies that interfere with serodiagnosis of infection, so that the serological diagnosis of vaccinated animals must be only realized after 24 months of age, when the vacinal antibodies declined to undetectable levels. The performance, kinetics and humoral clearance period of the strain 19 vaccine are well known in cattle, however there is little information concerning these aspects in buffalo. This study aimed to evaluate the elimination of strain S19 by buffalo females vaccinated with the standard dose containing 60-120 billion CFU, as well as the profile of the humoral immune response induced by vaccination and efficiency of principals laboratory tests to differentiate vaccinal antibodies of the antibodies induced by natural infection. We used 44 female buffalo from two farms negative for brucellosis, located in the State of São Paulo, being 33 animals of the property A and 11 of the property B. The animals were immunized and blood samples, urine and vaginal swabs were collected weekly from the time that vaccination is performed (D0) until complete 24 months of age. The sera obtained from the blood samples were used for serological testing, using the following tests: rose bengal test (RBT), 2-mercaptoethanol (2ME), complement fixation test (CFT) and fluorescence polarization assay (FPA). The urine samples and vaginal swabs were tested by microbiological culture for isolation of Brucella spp. and polymerase chain reaction (PCR) to evaluate the clearance of the vaccine strain. In PCR, the samples were first amplified using primers directed to the interspace region of the ribosomal RNA Brucella spp. (ITS-PCR) for the detection of any component of the genus. Samples with positive results by ITS-PCR were subjected to PCR using primers directed to the gene of ery of Brucella spp, in a specific assay for the detection of strain B19 (B19-PCR). Were calculated the specificity of each serological test to detect as negative the vaccinated animals. All animals showed reagents at seven days post-vaccination in the RBT test, at 14 days in the test 2ME, 126 on the CFT test. In the test of FPA, D28 is the period in which the highest number of animals tested positive. Sixteen animals do not seroconverted at any time of the experiment by FPA. The FPA test was more effective in differentiating antibodies of vaccination, and all animals were negative at 151 and 232 days post vaccination in the properties A and B, respectively. In test CFT, 100% animals showed negative results in D552 (property A) and D331 (property B). In the property A, at D552, the specificities were 92.31% for the test 2ME and 69.23% for the test RBT. Property B had in the days D366 specificities of 72.73% and 36.36% for the tests 2ME and RBT, respectively. All animals were negative by microbiological culture throughout the monitoring period. In the PCR were verified 19 samples positives in 1144 (1.66%). Of the 44 heifers evaluated, 15 (34.1%) showed positive results in at least one sample, in samples of urine and/or vaginal swabs. According to the presented results, we can conclude that the most effective serological test for differentiation of antibodies of natural infection of vaccine testing is FPA and the elimination of the vaccine strain may occur intermittently during periods prolonged in buffalo females vaccinated.