"Tesis para el doctorado. Facultad de medicina y ciencias naturales de la Universidad de Cartagena." / At head of title: Ismael Porto M.

This thesis describes the West Nile virus (WNV) epidemic in horses by exploring all aspects: sub-clinical infection, development of clinical disease and case fatality. All of the collected data were then compiled to create predictive risk maps of WNV infection for the province of Saskatchewan. <p>During the 2003 season, 133 clinical cases were documented with laboratory testing. Week of onset of clinical signs, gender, and coat color were significant predictors of whether the horse died or was euthanized due to severity of clinical signs. Studies of the serological response to vaccination and natural infection were examined to interpret the lab results from over 1100 samples taken from approximately 875 horses in 2003. A serologic study involving 212 horses on 20 farms determined the prevalence of sub-clinical infection (55.7% (95%CI, 44.9% to 65.8%)) and identified risk factors for infection. The study found risk of infection was highest in the Grasslands ecoregions compared to the Boreal Transition ecoregion. A case control study looked at risk factors for development of clinical disease. The study followed 23 case farms and control farms with a total of 300 horses sampled. This was the first field study to show that vaccination was efficacious in preventing the development of clinical signs. <p>The inclusion of horse surveillance data in the Saskatchewan Health WNV Integrated Surveillance Initiative was useful; however, it was discontinued due to time constraints, logistics, and declining monetary resources. <p>Since West Nile Virus is a mosquito-borne disease it is highly influenced by environmental changes, spatially and temporally. Discriminant analyses were used to partition Saskatchewan rural municipalities (RM) into categories of risk of infection with WNV based on acquired horse data and different environmental and meteorological data derived from both satellites or climate stations. The result was the creation of yearly predictive risk maps defining low to high risk of infection with WNV for each RM. <p>The 2003 epidemic provided a novel opportunity to study an important zoonotic disease emerging in a new environment. The information gathered will further the knowledge base upon which decisions for prevention of infection and clinical disease are made.

This thesis describes the West Nile virus (WNV) epidemic in horses by exploring all aspects: sub-clinical infection, development of clinical disease and case fatality. All of the collected data were then compiled to create predictive risk maps of WNV infection for the province of Saskatchewan. <p>During the 2003 season, 133 clinical cases were documented with laboratory testing. Week of onset of clinical signs, gender, and coat color were significant predictors of whether the horse died or was euthanized due to severity of clinical signs. Studies of the serological response to vaccination and natural infection were examined to interpret the lab results from over 1100 samples taken from approximately 875 horses in 2003. A serologic study involving 212 horses on 20 farms determined the prevalence of sub-clinical infection (55.7% (95%CI, 44.9% to 65.8%)) and identified risk factors for infection. The study found risk of infection was highest in the Grasslands ecoregions compared to the Boreal Transition ecoregion. A case control study looked at risk factors for development of clinical disease. The study followed 23 case farms and control farms with a total of 300 horses sampled. This was the first field study to show that vaccination was efficacious in preventing the development of clinical signs. <p>The inclusion of horse surveillance data in the Saskatchewan Health WNV Integrated Surveillance Initiative was useful; however, it was discontinued due to time constraints, logistics, and declining monetary resources. <p>Since West Nile Virus is a mosquito-borne disease it is highly influenced by environmental changes, spatially and temporally. Discriminant analyses were used to partition Saskatchewan rural municipalities (RM) into categories of risk of infection with WNV based on acquired horse data and different environmental and meteorological data derived from both satellites or climate stations. The result was the creation of yearly predictive risk maps defining low to high risk of infection with WNV for each RM. <p>The 2003 epidemic provided a novel opportunity to study an important zoonotic disease emerging in a new environment. The information gathered will further the knowledge base upon which decisions for prevention of infection and clinical disease are made.

Thesis (Ph.D. (Tropical Medicine))--Mahidol University, 2003.

Made available in DSpace on 2014-06-11T19:25:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-09-02Bitstream added on 2014-06-13T19:12:38Z : No. of bitstreams: 1 costa_tac_me_araca.pdf: 243307 bytes, checksum: 51ba4339b5f183921482d79e00370718 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O presente trabalho teve como objetivos estudar, em uma área endêmica para leishmaniose visceral, a soroprevalência anticorpos anti-Leishmania spp. em felinos, por meio duas técnicas de ensaio imunoenzimático (ELISA), ELISA-prot.A e ELISA-IgG, utilizando-se 200 gatos. Para avaliar a performance dos testes sorológicos utilizados no diagnóstico da leishmaniose felina, foram calculadas as especificidades e sensibilidades de cada teste, bem como o índice kappa (k) para cada um deles, com o intuito de avaliar a concordância dos métodos com o teste parasitológico direto (padrão ouro). Foram observadas formas amastigotas do patasito em 4% (8/200) gatos. Pelo ELISA-Prot.A, 4,5% (9/200) dos gatos apresentaram títulos acima do ponto de corte para a espécie e, pelo ELISA-IgG, 11,5% (23/200) dos animais foram considerados soropositivos. O ELISA-Prot.A apresentou sensibilidade de 12,5%, especificidade de 64,5% e concordância fraca com o teste parasitológico direto. O ELISA-IgG apresentou sensibilidade de 25% e especificidade de 19,2%, com índice kappa também indicando fraca concordância. / The present work aimed to study, in an endemic area for visceral leishmaniasis, the seroprevalence of Leishmania sp. infection in cats using two enzyme-linked immunosorbent assay (ELISA) techniques, ELISA-protein A and ELISAimmunoglobulin G. For this purpose, a total of 200 cats were employed. To evaluate the performance of the available sorological tests for the diagnosis of feline leishmaniasis it was determined and compared the sensitivity and specificity of ELISA-prot. A and ELISA-IgG, as well as the kappa index (k) for each one, to measure its concordance with the parasitilogical test (gold standard). Amastigote forms of the parasite were observed in eight (4.0%) cats. By ELISA-proteín A nine (4,5%) cats presented titer above the specie’s cut off point and by ELISA-IgG 23 (11,5%) were considered seropositive. The ELISA-proteín A was 12,5% sensitive and 65,4% specific, and showed a weak concordance with the parasitological test. The ELISA-IgG showed a sensitivity of 25% and specificity of 19,2% and when kappa index was analysed, a weak concordance was observed too.

Orientador: Mary Marcondes / Banca: Valéria Marçal Felix de Lima / Banca: Márcia Dalastra Laurenti / Resumo: O presente trabalho teve como objetivos estudar, em uma área endêmica para leishmaniose visceral, a soroprevalência anticorpos anti-Leishmania spp. em felinos, por meio duas técnicas de ensaio imunoenzimático (ELISA), ELISA-prot.A e ELISA-IgG, utilizando-se 200 gatos. Para avaliar a performance dos testes sorológicos utilizados no diagnóstico da leishmaniose felina, foram calculadas as especificidades e sensibilidades de cada teste, bem como o índice kappa (k) para cada um deles, com o intuito de avaliar a concordância dos métodos com o teste parasitológico direto (padrão ouro). Foram observadas formas amastigotas do patasito em 4% (8/200) gatos. Pelo ELISA-Prot.A, 4,5% (9/200) dos gatos apresentaram títulos acima do ponto de corte para a espécie e, pelo ELISA-IgG, 11,5% (23/200) dos animais foram considerados soropositivos. O ELISA-Prot.A apresentou sensibilidade de 12,5%, especificidade de 64,5% e concordância fraca com o teste parasitológico direto. O ELISA-IgG apresentou sensibilidade de 25% e especificidade de 19,2%, com índice kappa também indicando fraca concordância. / Abstract: The present work aimed to study, in an endemic area for visceral leishmaniasis, the seroprevalence of Leishmania sp. infection in cats using two enzyme-linked immunosorbent assay (ELISA) techniques, ELISA-protein A and ELISAimmunoglobulin G. For this purpose, a total of 200 cats were employed. To evaluate the performance of the available sorological tests for the diagnosis of feline leishmaniasis it was determined and compared the sensitivity and specificity of ELISA-prot. A and ELISA-IgG, as well as the kappa index (k) for each one, to measure its concordance with the parasitilogical test (gold standard). Amastigote forms of the parasite were observed in eight (4.0%) cats. By ELISA-proteín A nine (4,5%) cats presented titer above the specie's cut off point and by ELISA-IgG 23 (11,5%) were considered seropositive. The ELISA-proteín A was 12,5% sensitive and 65,4% specific, and showed a weak concordance with the parasitological test. The ELISA-IgG showed a sensitivity of 25% and specificity of 19,2% and when kappa index was analysed, a weak concordance was observed too. / Mestre

A ocorrência de Listeria monocytogenes em alimentos vem sendo estudada desde o início dos anos 80, após seu envolvimento em vários surtos de doença de origem alimentar. Os frutos do mar são o grupo de alimentos que despertou menor atenção por parte dos pesquisadores, apesar de terem sido envolvidos em casos esporádicos de listeriose e mesmo em surtos da doença. A amostragem ambiental e de produto, ao longo de uma linha de processamento é uma forma de localizar áreas relacionadas à contaminação do alimento permitindo que sejam feitas correções para evitar a produção de bens que exponham o consumidor a doenças. Com a finalidade de avaliar a contribuição da matéria prima e fatores ambientais na ocorrência e distribuição de L. monocytogenes em uma indústria processadora de pescados, e mais especificamente, numa linha processadora de camarão rosa (penoeus brasiliensis), é que desenvolveu-se a presente pesquisa. Também buscou-se determinar as diversidades antigênica e genética das cepas de L. monocytogenes isoladas, e correlacionar esta diversidade à sua distribuição na indústria. Assim, um total de 363 amostras coletadas em diferentes pontos de uma linha de processamento de camarão rosa foram examinadas para a presença de L. monocytogenes, empregando-se a metodologia recomendada pelo Health Protection Branch, do Canadá. A seguir, 115 cepas de L. monocytogenes representativas das amostras positivas para o microrganismo foram sorotipadas e o polimorfismo do seu DNA cromossomico avaliado com o auxílio dos métodos RAPD (\"randon amplified polimorphic DNA\") e PFGE (\"pulsed-field MTDestro - Doutorado - Listeria monocytogeneses em camarao... 140 gel electrophoresis\"). Um grupo de 25 cepas foi também submetido a sorotipagem completa, fagotipagem e ribotipagem pelo sistema RiboPrint da DuPont. Do total de amostras examinadas, 64 (17,6%) apresentaram-se contaminadas por L. monocyfogenes. As amostras ambientais apresentaram 25,0% de positividade (14 positivas/56 examinadas). As amostras de utensílios 24,2% (8/33) e as de água 23,8% (5/21). As amostras de camarão apresentaram 18,0% de contaminação (32/178) e as de manipuladores 7,6% (5/66). Nenhuma das amostras de gelo foi positiva para L. monocyfogenes. Durante o processamento observou-se um aumento na percentagem de amostras positivas para L. monocytogenes, chegando esta percentagem a 35,0% em algumas etapas, mas reduzindo-se a 16,0% no produto final. O perfil composto gerado pela combinação dos resultados obtidos com a tipagem molecular das 115 cepas de L. monocyfogenes selecionadas, permitiu a sua divisão em 24 grupos, de acordo com seus perfis de DNA. A sorotipagem das 25 cepas selecionadas mostrou que 11 pertenciam ao sorovar 4b, 7 ao sorovar 1/2b, 2 ao sorovar 1/2c e 5 ao sorovar 1/2a. A fagotipagem permitiu que elas fossem divididas em 7 fagovares, sendo que a maior parte das cepas do sorovar 4b (81,8%) não foi fagotipável. A ribotipagem destas 25 cepas originou 6 RiboGroupsTM. Os resultados indicaram que cepas de L. monocytogenes de origem ambiental pertenciam a perfis compostos exclusivos para o ambiente, enquanto que cepas provenientes da água e de utensílios possuiam um perfil composto em comum. Amostras de camarão coletadas nas diversas etapas doprocessamento apresentaram L. monocytogenes com pelo menos um perfil composto em comum, perfil este também presente nas cepas isoladas das mãos dos manipuladores. / The importance of seafood in the spread of foodborne pathogens is well known, however, until the last few years, little attention has been paid to the role of seafood in disseminating L. monocytogenes. Two foodborne listeriosis outbreaks have been linked to the consumption of seafood. L. monocytogenes and other Listeria species have been isolated from different types of raw or processed seafood, but the main source of contamination is unknown. For this reason, it is important to monitor the potential sources of this pathogen in food processing plants, in order to minimize product contamination. The aim of this study was to evaluate the contribution of raw material and environment in the occurrence and distribution of L. monocytogenesin shrimp (Penaeus brasiliensis) processing plant. The antigenic and genetic diversities of L. monocytogenes strains were also determined. A total of 363 samples, collected in different areas of a shrimp processing plant in Santos, SP, were examined using the methodology recomended by the Health Protection Branch, Canada. One hundred and fifteen strains of L. monocytogenes representing the L. monocytogenes positive samples were first serotyped and then sub-typed by molecular typing (RAPD and PFGE). A group of 25 strains were also ribotyped and phage-typed. L. monocytogeneswas isolated from 64 (17.6%) of the total samples analysed. Environmental samples showed the highest positivity rate (25%) followed by utensils (24,2%) and water (23.8%) samples. Shrimp samples presented 18% of positivity for L. monocytogenes while food handlers samples presented 7.6%. None of the ice samples was positive for the microorganism. When the composite profile from \"both (RAPD-PFGE) methods was generated, the 115 strains could be separated in 24 groups, according to their DNA pattern. The results indicated that environmental strains of L. monocytogenes ali feel into composite profile groupings unique to the environment, while strains from both water and utensils shared another composite profile group. L. monocytogenes fresh shrimp iso/ates belonging to one profile group, were found in the different areas of the processing line. This same latter group was also present in food handlers from the processing and packaging areas of the plant.

As hantaviroses são zoonoses de roedores silvestres que causam doenças humanas graves: febre hemorrágica com síndrome renal na Ásia e Europa, com letalidade de 10 a 12% e no continente americano causam a Síndrome Pulmonar e Cardiovascular por Hantavirus (SPCVH), letalidade de 59%. Os Hantavirus (família Bunyaviridae) são vírus envelopados que medem aproximadamente 120 nm, possuem RNA de fita simples e polaridade negativa, dividida em 3 segmentos (L, M e S) que se replicam no citoplasma. A infecção humana relaciona-se à inalação de aerossóis de excretas de roedores infectados com Hantavirus, embora existam relatos de transmissão interpessoal na América do Sul. No Estado de São Paulo, os roedores infectados mais encontrados foram o “rato do rabo peludo" (Bolomys lasiurus), o “rato da mata" (Akodon cursor) e o “ratinho do arroz" (Oligoryzomys negrips). Desde os 3 primeiros casos de hantavirose descritos no Brasil (1993) com indivíduos moradores da área rural de Juquitiba, SP, evidenciando o primeiro surto conhecido de SPCVH, mais de uma centena de casos da SPCVH foram notificados, causando 60 óbitos (letalidade de 46%). Na região de Ribeirão Preto, ocorreram quatorze casos de SPCVH de 1998 a 2001, com letalidade de 64,2%, o que motivou este trabalho. Um primeiro objetivo deste trabalho foi estudar aspectos clínicos e epidemiológicos dos casos de hantavirose ocorridos na região de Ribeirão Preto, de 1998 a 2001, para conhecer o comportamento das hantaviroses nesta região. Analisando e acompanhando prontuários de 14 pacientes com SPCVH, observou-se febre (100% dos casos), estertores pulmonares, dispnéia e tosse (64,2%), taquicardia acompanhada de hipotensão (64,2%), cefaléia e sintomas digestivos (57%), adinamia e indisposição geral (50%), fenômenos hemorrágicos (28,5%), mialgia (21,4%) e convulsão (7,5%). No perfil laboratorial, a plaquetopenia <130000/mm3 (100%), hematócrito >55%, (78,6%); leucocitose >15000/mm3 (64,2%), neutrofilia >7000/ mm3 e bastonetes >600 cels/mm3 (64,2%), creatinina sérica >1,5mg/100ml (63,6%) e PO2 <70 mmHg (54,5%). O diagnóstico laboratorial das hantaviroses foi feito por ELISA para IgM e IgG anti Sin Nombre após a alta. Houve diferença significativa entre casos fatais e sobreviventes, quanto à época de suspeita diagnóstica de hantavirose (p=0,0152) e quanto ao uso de hidratação parenteral (p=0,0152): nos sobreviventes, a suspeita diagnóstica de SPCVH foi feita entre o 1º e o 2º dia de internação e, nos casos fatais, a suspeita foi feita apenas na necropsia. Existe uma correlação significativa entre o volume de infusão endovenosa de líquidos &#61619; 2500 ml, nas primeiras 24h de tratamento, e a evolução dos casos para o óbito. A oxigenação precoce, ventilação mecânica, assim como o uso de aminas vasoativas e o de corticosteróides não tiveram associação com sobrevida e a presença de choque não teve associação com óbito. O segundo objetivo foi estudar a prevalência de anticorpos para Hantavirus na população de Jardinópolis, na região de Ribeirão Preto, SP. Em estudo prévio entre cidades da região de Ribeirão Preto, obteve-se maior índice de prevalência sorológica (4,5%) para Hantavirus naquele município. Através de coleta aleatória em parte da área rural e em toda a área urbana, obtivemos 818 amostras de sangue de moradores entre 15 e 70 anos, por digitopuntura em papel de filtro, dos quais coletamos dados relevantes como idade, sexo, cor, procedência, atividade profissional, endereço, telefone; moléstia atual, medicamentos em uso, pneumonia grave pregressa, tipo de moradia, número de moradores na casa, esgoto, coleta de lixo, celeiro, contato com roedores. Essas amostras foram processadas nas diluições 1/50, 1/100, 1/400 e 1/1600 e submetidas ao ELISA indireto para detecção de IgG para Hantavirus Andes. Os resultados foram analisados pelo teste de Fisher e teste do Qui-quadrado, utilizando o programa InStat 3.0 (GraphPad Software Inc, USA) e para &#945; de 5%, com intervalo de confiança de 95%. A positividade geral foi de 14,3% com ELISA à diluição 1/50, na zona urbana foi de 15,3% e na zona rural de 6,5%, demonstrando alta representatividade para a população do município de Jardinópolis. Entre moradores de zona urbana e rural, a positividade para Hantavirus foi significativamente maior para os moradores urbanos (p= 0,0183). E entre as diferentes faixas etárias com a das restantes, observou-se diminuição significativa para a faixa etária de 21 a 30 anos (p= 0,0117) e aumentada para a de 31 a 40 anos (p= 0,0036). Não houve diferença significativa de positividade para Hantavirus observada nos homens e nas mulheres (p= 0,0892), nem para indivíduos com atividade profissional urbana e rural (p=0,8799), nem com os indivíduos que referiram pneumonia grave (p= 0,6096) e da mesma forma nos indivíduos que declararam contato com roedores (p= 0,4842). Assim como não houve associação entre títulos de 100 e 400 para as mesmas características analisadas. / The hantaviruses are zoonoses of wild rodents that cause serious human diseases: hemorrhagic fever with renal syndrome in Asia and Europe (10 to 12% lethality rates) and Pulmonary and Cardiovascular Syndrome in the American continent (HPCVS - 59% lethality rate). Hantaviruses (family Bunyaviridae) are enclosing viruses that measure 120 nm approximately, they possess RNA of simple ribbon and negative sense, divided in 3 segments (L, M and S) that are replication in the cytoplasm. The human infection links to the inhalation of aerosols of Hantavirus infected rodents excretes, although reports of person-to-person transmission exist in South America. In the State of São Paulo the infected rodents found were the Bolomys lasiurus, the Akodon cursor and the Oligoryzomys negrips. From the first 3 cases of hantaviruses described in Brazil (1993) with individuals inhabitants of the rural area of Juquitiba, SP, evidencing the first well-known supplies of HPCVS, more than a hundred cases have been notified in Brazil, causing 60 deaths (of 46% lethality rate). There have been fourteen cases of HPCVS since1998 to 2001 with of 64,2% letality rate, in the Ribeirão Preto area what motivated this work. One of the objectives was clinical and epidemic aspects studies of the cases of hantaviruses that happened in the Area of Ribeirão Preto, from 1998 to 2001, to know the behavior of the hantavirusess in this area. Analyzing and accompanying patient medical records of 14 patients with HPCVS, fever was observed (100% of the cases), disnea and coughs (64,2%), accompanied of arterial hipotension, tachicardia (64,2%), migraine and digestive symptoms (57%); adynamic and general weakness (50%), hemorrhagic phenomenons (28,5%), mialgia (21,4%) and convulsion (7,5%). In the laboratorial profile the platelets count <130000/mm3 (100%), hematocrit> 55%, (78,6%); leucocytosis > 15000/mm3 (64,2%), neutrofilia >7000 / mm3 and small sticks> 600 cels / mm3 (64,2%), creatinine serical level > 1,5mg/100ml (63,6%) and PO2 <70 mmHg (54,5%). The diagnosis laboratorial of the hantaviruses was made by ELISA for IgM and IgG anti Sin Nombre after the discharge. There was significant difference among fatal and surviving cases, as the hantaviruses diagnosis suspicious (p=0,0152) and parenteral hydratation (p=0,0152): in the survivors, the HPCVS diagnosis suspicious was made between the 1st and the 2nd day of internment and the fatal cases, the suspicion was just made to the necropsies. Among infusion of liquid EV ³ over 2500 ml in the first 24 hours of treatment and evolution for the death. The precocious oxygen support, mechanics ventilation, as well as the use of vasoatives amines and the one of corticosteróides, did not have association with survival and the shock presence did not have association with death. The second objective was to antibodies for Hantavirus prevalence studies in the Area of Ribeirão Preto, in the population of Jardinópolis, SP. In the previous study among cities in that area it was obtained larger index of serologic prevalence (4,5%) for Hantavirus, it was in the Jardinópolis county Through ramdomised collection partly of the rural area and whole urban area, we obtained 818 samples of blood, inhabitants between 15 and 70 years, for digital puncture in filter paper and important data as age, gender, color, origin, professional activity, address, telephone; current disease, medications in use, pneumonia serious, dwelling type, number of inhabitants in the house, sewer, collects of garbage, barn, contact with rodents. These samples were processed, in the dilutions 1/50, 1/100, 1/400 and 1/1600. and submitted to indirect ELISA for detection of IgG for Hantavirus Los Andes. The results were analyzed by Fisher and Qui-square tests, using the InStat 3.0 program (GraphPad.Software Inc, USA) and for the one of 5%, with interval of trust of 95%. The general positivity rated 14,3% with ELISA (at title 50), urban zone (15,3%) and in rural zone 6,5% and they demonstrated high rates for the population of the Jardinópolis county. Comparing the positivity rates for Hantavirus between urban and rural zone inhabitants was significantly larger for the urban inhabitants (p = 0,0183). Significant increase was observed for the age group of 21 to 30 years (p = 0,0117) and for the one of 31 to 40 years (p = 0,0036) among different age groups. There was not significant difference of positivist for Hantavirus between men and women (p = 0,0892); nor for individuals with urban and rural professional activity (p=0,8799), neither for the individuals that referred serious pneumonia (p = 0,6096) as well as in the individuals that declared contact with rodents (p = 0,4842). There was not association among titles of 100 and 400 for Hantavirus with gender, local of habitation, contact with rodents and pneumonia serious.

Thesis (Ph.D. (Tropical Medicine))--Mahidol University, 2003.

Thesis (M.A.)--University of South Florida, 2006. / Title from PDF of title page. Document formatted into pages; contains 86 pages. Includes bibliographical references.