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Avaliou-se o impacto da fragmentação florestal na diversidade de carrapatos de vida-livre e a presença de patógenos nestes carrapatos em remanescentes florestais do Pontal do Paranapanema, estado de São Paulo, Brasil. Estes fragmentos florestais abrigam uma rica e importante biodiversidade, com espécies endêmicas e ameaçadas como o mico-leão-preto (Leontopithecus chrysopygus), a anta (Tapirus terrestris), a onça pintada (Panthera onca), o macuco (Tinamus solitarius) e várias outras espécies de aves, mamíferos, répteis, anfíbios e peixes. Aproximadamente 90% das espécies de carrapatos parasitam exclusivamente hospedeiros selvagens. O restante pode ter animais domésticos e humanos como hospedeiros. Embora a maioria das pesquisas tenha sido dirigida a espécies de importância econômica, os carrapatos que parasitam animais selvagens possuem relevante papel na manutenção dos níveis de patógenos em populações de vida livre. Algumas destas espécies, por exemplo, demonstraram que podem parasitar hospedeiros não selvagens e promover o surgimento de zoonoses. Em habitats fragmentados, a diversidade de espécies vertebradas é menor se comparado a habitats com pouca alteração antrópica. Portanto, a fragmentação do habitat diminui a diversidade de espécies de carrapatos também. Para estudar a relação entre a fragmentação florestal e a ecologia das populações de carrapatos foram coletados carrapatos em 8 fragmentos florestais pelo método de arrasto de flanela e inspeção visual da vegetação. Os índices de comparação utilizados foram de similaridade de Jaccard, de diversidade de Shannon e complexidade do fragmento florestal de Patton. Utilizou-se o modelo de regressão linear para compara os índices de Shannon e Patton. Um total de 2149 ninfas de Amblyomma spp foi coletado e foram identificadas as espécies de 629 carrapatos. As espécies coletadas foram Amblyomma cajennense (94,28%), A. coelebs (1,59%), A. naponense (2,86%), A. ovale (0,64%), A. nodosum (0,32%), A. brasiliense (0,16%) e Haemaphysalis juxtakochi (0,16%). Nenhum indivíduo testado foi positivo pelo teste da hemolinfa. Os resultados mostraram uma tendência de correlação entre a fragmentação floresta, e a diversidade de espécies de carrapatos. / This study evaluated the impact of forest fragmentation on diversity of freeliving ticks and prevalence of tick pathogens in remaining forest fragments in the Pontal do Paranapanema, São Paulo state, Brazil. These forest fragments shelter rich and important biodiversity, with endemic and threatened species such as the black lion tamarin (Leontopithecus chrysopygus), the tapir (Tapirus terrestris), jaguar (Panthera onca), the solitary tinamou (Tinamus solitarius) and various other species of birds, mammals, reptiles, amphibians and fish. Approximately 90% of tick species parasitize exclusively wild hosts. The remainder can also have domestic animals and humans as hosts. Although most research has been directed to species of economic importance, ticks that parasitize wild animals are also relevant due to their role in maintaining enzootic pathogen levels in wild populations. Some of these species, for example, have been shown to cross-over onto non-wild hosts and promote emergent zoonoses. In fragmented habitats, the diversity of vertebrate species is normally lower than comparable habitats with minimal anthropic alteration. Thus, habitat fragmentation decreases the diversity of tick species too. To study the relationship between forest fragmentation and population ecology of ticks, ticks were collected in 8 forest fragments using dragging and visual inspection of vegetation. The index used were Jaccard´s similarity, diversity f Shannon and Patton. The linear regression model was used to compare Shannon and Patton indexes. A total of 2149 nymphs of Amblyomma spp. And 629 identified ticks was collected. The species of ticks collected was Amblyomma cajennense (94,28%), A. coelebs (1,59%), A. naponense (2,86%), A. ovale (0,64%), A. nodosum (0,32%), A. brasiliense (0,16%) e Haemaphysalis juxtakochi (0,16%). All ticks were negative by hemolimph test. The results showed a tendency of correlation between forest fragmentation and diversity of tick species.

Anseriformes mantidos em lagos de zôos e parques estão sob constante risco de exposição às doenças presentes nas populações de aves migratórias, que dividem com eles o mesmo local durante um determinado período todos os anos. São doenças que podem ter implicações para as aves cativas, para a população humana que tem contato com essas aves e para os plantéis de produção. Assim, ações de vigilância, com o objetivo de detectar rapidamente determinadas doenças, representam alternativas interessantes para se fazer gestão de risco. O objetivo do presente estudo foi pesquisar a presença de agentes etiológicos selecionados na população de Cisnes Negros (Cygnus atratus), mantida nos lagos da FPZSP, visto que essa população tem contato com as seguintes aves migrantes que visitam a FPZSP todos os anos: irerês (Dendrocygna viduata), marreca caneleira (Dendrocygna bicolor) e marreca asa de seda (Amazoneta brasiliensis). Assim, foram colhidos suabes de traquéia e cloaca de uma amostra capaz de detectar doença com prevalência estimada em 1% para um nível de confiança de 95%. Além disso, foi realizado um estudo retrospectivo (2001 a 2006) das principais causas de morte nessa população. As principais causas de mortalidade registradas em 184 registros analisados foram: desvio de tendão extensor tarso-metatarsiano (37, 20,1%), desnutrição (20, 10,9%), problemas hepáticos (17, 9,2%), traumas (15, 8,2%), problemas respiratórios (8, 4,3%), septicemias (6, 3,3%), intoxicações (5, 2,7%) e problemas gastrointestinais (3, 1,6%). Um terço das carcaças (62, 33,7%) foi encontrado em estado de putrefação. A taxa de mortalidade foi decrescente de 2001 a 2006 e apresentou sazonalidade, sendo maior entre os meses de novembro a maio. No momento das coletas, não houve nenhuma evidência clínica ou laboratorial da presença dos seguintes agentes: Pasteurella multocida., Salmonella sp., Chlamydophila psittaci, Orthomixovírus (Influenza Aviária), Paramixovirus (Doença de Newcastle) e Coronavirus (Bronquite Infecciosa). / Waterfowls housed in ponds of zoos and parks are under constant risk of exposure to pathogens of migratory birds that visit these places every year. Some of them involving zoo animals and humans. The spread of particular diseases may also become a serious threat for domestic poultry. So, surveillance, focused in early detection of some diseases, can be an interesting tool to do risk management. The goal of the present work was to search the presence of some select pathogens in the captive black swan population (Cygnus atratus) present in the ponds of the Fundação Parque Zoológico de São Paulo (FPZSP), because these animals have contact with the following free-living waterfowls: white-faced whistling-duck (Dendrocygna viduata), fulvous whistling-duck (Dendrocygna bicolor) e brasilian teal (Amazoneta brasiliensis). Swabs of trachea and cloaca were sampled from 239 animals, the sample size required for detecting disease present in at least 1% of the animals (CI = 95%). Moreover, a retrospective study was done about the causes of death to the period from 2001 to 2006. The mainly causes of black swan death in FPZSP were: slipped tendon (37/184, 20,1%), malnutrition (20/184, 10,9%), hepatic problem (17/184, 9,2%), trauma (15/184, 8,2%), respiratory problem (8/184, 4,3%), septicemias (6/184, 3,3%), intoxication (5/184, 2,7%) e gastro-intestinal problems(3/184, 1,6%). One third of the carcass (62/184, 33,7%) was in autolysis. The mortality presented peaks of occurrence from november to may and a decreasing trend from 2001 to 2006. At the moment of the sampling, there was no clinical or laboratorial evidence of the infection by the following pathogens: Pasteurella multocida., Salmonella sp., Chlamydophila psittaci, Orthomixovírus (Avian Influenza), Paramixovirus (Newcastle Disease) e Coronavirus (Infectious Bronchitis).

Foram coletados cinqüenta e oito (58) amostras de cachorro-do-mato (Cerdocyon thous) de vida livre, provenientes do Estado do Espírito Santo, Brasil. Os animais eram mortos por atropelamento ao longo da rodovia estadual ES-060 que liga os municípios de Vila Velha à Guarapari, passando por duas reservas florestais de Mata Atlântica. Todos os animais eram encaminhados para a Universidade de Vila Velha UVV pela concessionária RodoSol, onde passavam por detalhado exame necroscópico com coleta de tecidos e ectoparasitas para pesquisa de patógenos pela técnica de Reação em Cadeia de Polimerase (PCR). Todas as amostras obtidas foram testadas quanto à presença de agentes pertencentes à família Anaplasmataceae e membros dos gêneros Rickettsia, Borrelia, Babesia, Coxiella, Leishmania, Hepatozoon e Ehrlichia. No total foram colhidos 25 espécimes de carrapatos, todos em estágio ninfal e pertencentes à espécie Amblyomma cajennense. Na pesquisa de patógenos pela PCR, não foi encontrada nenhuma amostra de animal ou carrapato positiva para Leishmania spp, Rickettsia spp, Borrelia spp, Babesia spp e Coxiella spp. Das 58 amostras de tecidos, 29 (50%) foram positivas para Hepatozoon spp no gene 18S rRNA, tendo sido identificado dois genótipos, um denominado Hepatozoon sp. ex Cerdocyon thous, presente em 96,55% dos animais testados, com máxima similaridade de 98,67% com a espécie Hepatozoon sp. curupira 2 (AY461377). O outro genótipo foi encontrado em somente um animal (3,44%), denominado Hepatozoon sp. P20 Cerdocyon thous, que apresentou máxima similaridade de 97,5% com Hepatozoon sp. 744C (EU430234). Na pesquisa para Ehrlichia spp, seis amostras foram positivas (10,34%). As seis amostras foram caracterizadas como uma possível nova espécie de Ehrlichia, denominada Ehrlichia sp. ex Cerdocyon thous, com máxima similaridade de 97,57% com a espécie Ehrlichia ruminantium (DQ482915) para o gene 16S e 82,51% similar com E. ruminantium str. Gardel (CR925677) para o gene dsb. Todas as espécies de carrapatos foram negativas. Hepatozoon spp e Ehrlichia spp são agentes infecciosos transmitidos por carrapatos, com potencial zoonótico, de potencial impacto para saúde animal e humana. / We collected samples from fifty-eight (58) free-living crab-eating fox (Cerdocyon thous) from the State of Espírito Santo, Brazil. All animals were hit by vehicles passing through the highway ES-060, which crosses an Atlantic rainforest reserve, located between the cities of Guarapari and Vila Velha. The animals were collected by the Rodosol Company, and taken to the University of Vila Velha - UVV, where they underwent post-mortem examination, and collection of tissue samples and ectoparasites for research through the polymerase chain reaction (PCR) technique. All samples were tested for the presence of agents belonging to the family Anaplasmataceae and members of the genera Rickettsia, Borrelia, Babesia, Coxiella, Leishmania, Hepatozoon, and Ehrlichia. We collected 25 specimens of nymphal ticks, which were all identified as Amblyomma cajennense. Search of pathogens by PCR did not find any positive animal or tick sample for Leishmania spp, Rickettsia spp, Borrelia spp, Babesia spp and Coxiella spp. From the 58 tissue samples, 29 (50%) were positive for Hepatozoon sp by the PCR targeting the 18S rRNA gene. Amplicons generated two different genotypes, one named Hepatozoon sp. ex Cerdocyon thous, present in 96.55% of the animals, with a maximum of 98.67% similarity with the sequence of Hepatozoon sp. curupira 2 (AY461377); the second genotype was found in only one animal (3.44%), named Hepatozoon sp. P20 Cerdocyon thous, which showed maximum similarity (97.5%) with Hepatozoon sp. 744C (EU430234). The survey for Ehrlichia spp resulted in six positive samples (10.34%). The six samples were characterized as a possible new species of Ehrlichia, named Ehrlichia sp. ex Cerdocyon thous, with maximum similarity (97.57%) with the species Ehrlichia ruminantium (DQ482915) for the 16S and 82.51% similar to E. ruminantium str. Gardel (CR925677) for the dsb gene. All species of ticks were negative for all pathogens searched. Hepatozoon spp and Ehrlichia spp are tick-borne infectious agents with zoonotic potential; the present findings are of potential impact for both animal and human health.

Introdução: Os vírus parainfluenza são importante causa de infecções respiratórias. Na população pediátrica a doença característica associada com HPIV-1 e 2 é a laringotraqueobronquite, enquanto que o HPIV-3 está associado com a pneumonia e bronquiolite. Objetivos: Realizar a análise molecular do fragmento do gene da HN. Métodos: Foram analisados aspirados nasofaríngeos coletados entre 1995 a 2005, de crianças com doença respiratória aguda. A detecção dos HPIV 1, 2 e 3 foi realizada por multiplex RT-PCR. Posteriormente, o fragmento de gene HN foi amplificado, sequenciado e, a análise molecular foi realizada. Resultados e discussão: Um total de 6% amostras foram positivas para HPIV, sendo o HPIV-3 o vírus mais prevalente durante todos os anos estudados, o que corresponde a 80% dos casos positivos, seguido por HPIV-1 (15%) e, HPIV-2 (7%). A maioria das mutações observadas foram silênciosas, no entanto, algumas alterações de aminoácidos foram verificadas em áreas conservadas dos HPIV-3 e HPIV-2, e alterações em sítios potencias de N-glicosilação também foram observados. / Introduction: In paediatric population the characteristic illness associated with HPIV-1 and -2 is laryngotracheobronchitis, whereas HPIV-3 is associated with pneumonia and bronchiolitis. There are 5 virus distributed in two distinct genus: Respirovirus (HPIVI-1 and HPIV-3) and Rubulavirus (HPIV-2, HPIVI-4A and HIVI-4B). Objectives: To carry out the molecular analysis of the fragment of the HN gene. Methods: Nasopharyngeal aspirates from infants with acute respiratory illness collected from 1995 to 2005, were analyzed. The detection of HPIV 1, 2 and 3 were performed by multiplex RT-PCR and the fragment of HN gene was amplified, sequenced and, the molecular analysis was carried out. Results and discussion: A total of 6% samples were positive for HPIV. The HPIV-3 was the most prevalent virus during, corresponding to 80% of positive cases, followed by HPIV-1 with 15% and HPIV-2 7%. Most mutations observed were silent in all PIVs, however, some amino acids alterations in conserved areas, verified in PIV-3 and PIV-2, and alterations in N-glicosilation sites were observed.

Atualmente, milhões de pessoas, por todo o globo, estão sob risco de serem infectados por um protozoário transmitido vetorialmente por pequenos insetos flebotomíneos. Este parasita é a Leishmania spp., causadora de uma patologia de amplo espectro, que varia desde a moléstia cutânea (tegumentar) até a visceral (kala-azar). A leishmaniose cutânea é a manifestação clínica de maior ocorrência (mais de 90% dos casos). A radiação ionizante, gerada em fonte de 60Co, tem sido utilizada com sucesso para promover alterações físico-químicas em diferentes protozoários, incluindo a Leishmania spp. Em trabalhos anteriores determinou-se que formas promastigotas de Leishmania amazonensis, irradiadas com diferentes doses de radiação gama, perdem sua viabilidade mantendo, porém, sua imunogenicidade. No presente trabalho, estudouse a utilização da radiação ionizante como ferramenta na seleção de formas metacíclicas do parasita em cultura axênica para a possível produção de imunógenos irradiados mais eficientes. Os resultados demonstram que culturas irradiadas com 400 Gy de radiação gama, possuem uma concentração de aproximadamente 75% de parasitas metacíclicos, capazes de produzir, in vitro, uma infecção que mimetiza a ocorrida naturalmente. Estes parasitas irradiados têm sua estrutura celular interna modificada mantendo, entretanto, seu arcabouço externo intacto. Camundongos de uma linhagem suscetível imunizados com leishmanias irradiadas com diferentes doses tiveram sua produção de imunoglobulinas aumentada, e mantiveram os títulos elevados após o desafio com parasitas não irradiados. Em outras linhagens pesquisadas este padrão se manteve, porém em títulos menores, sendo que camundongos imunodeficientes não responderam à imunização nem ao desafio. / Actually, millions of people around the globe are under the risk of infection by a protozoan transmitted by a bit of a sand fly. This parasite is a Leishmania spp. This causes a wide spectrum disease, since a coetaneous disease to a visceral one. The coetaneous form is the major clinical manifestation (above 90%). The ionizing radiation, produced in a 60Co font, had being successes used to promote physical-chemical transformations on different protozoans, including Leishmania spp. In previous work was determined that promastigotes forms of Leishmania amazonensis, irradiated with different doses of radiation, lost their viability maintaining, however, their immunogenicity. In this work, was studied the use of ionizing radiation as a tool for selection of metaciclic forms of the parasite in axenic culture, for a possible efficient irradiated immunogen production. Our results shown that cultures irradiated with 400 Gy of gamma irradiation, has 75% of metaciclic form, which are capable to produce, in vitro, an infection that is similar the natural occurrence. These irradiated parasites have their internal cellular structure modified, maintaining their external structure intact. Susceptible strain of mice immunized with leishmania irradiated with different doses had high immunoglobulin production, and maintained this production after the challenge with naive parasites. In other strains this default was similar, however in lower titles. Immunodeficient mice didnt produce immunoglobulin nor on the immunization or on the challenge.

Philosophiae Doctor - PhD / Bionomics of vector-borne pathogens (VBPs) is a complex phenomenon that involves understanding the ecology of arthropod borne pathogens and vertebrate hosts potentially involved in their transmission cycles. Investigations into the bionomics of viral and bacterial VBPs circulating in Baringo and Homa Bay Counties of Kenya were carried out. Specifically, vertebrate hosts represented in mosquito bloodmeals, presence of arboviruses in blood fed mosquitoes and patients presenting with acute undiagnosed febrile illnesses in rural health facilities, and tick borne pathogens (TBPs) diversity in ticks of animals were identified. Mosquitoes were trapped by BG sentinel and CDC light traps, while ticks were sampled directly from domestic animals and tortoises close to human habitation along the shores and adjacent islands of Lakes Victoria and Baringo in Kenya. Blood and sera were also sampled from patients presenting with acute febrile illnesses visiting four rural health facilities in Homa Bay County. Mosquitoes and ticks were sorted and identified to species using standard morphological taxonomic keys. All the biological samples (blood-fed mosquitoes, ticks and blood/sera) were processed using molecular and culture procedures for detection of VBPs (arboviruses, Ehrlichia, Anaplasma, Rickettsia and protozoa). Among 445 blood-fed Aedeomyia, Aedes, Anopheles, Culex, Mansonia, and Mimomyia mosquitoes, 33 bloodmeal hosts were identified including humans, eight domestic animal species, six peridomestic animal species and 18 wildlife species. Further detection of Sindbis and Bunyamwera viruses was done on blood-fed mosquito homogenates by Vero cell culture and RTPCR in Culex, Aedeomyia, Anopheles and Mansonia mosquitoes from Baringo that had fed on humans and livestock. In TBPs assay, 585 tick pools were analysed consisting of 4,126 ticks collected in both study areas. More ticks were sampled in Baringo (80.5%), compared to Homa Bay (19.5%). In Baringo, agents of ehrlichiosis were detected from Amblyomma and Rhipicephalus ticks including Ehrlichia ruminantium (12.3%), Ehrichia canis (10.5%) and Paracoccus sp. (4.4%). Agents of anaplasmosis included Anaplasma ovis (7.2%), Anaplasma platys (4.4%) and Anaplasma bovis (4.0%), all from Hyalomma, Amblyomma and Rhipicephalus ticks, as well as agents of rickettsiosis, including Rickettsia africae, Rickettsia aeschlimannii, Rickettsia rhipicephali, Rickettsia montanensis and a Rickettsia sp. that was not conclusively characterized. Babesia caballi, Theileria sp. and Hepatozoon fitzsimonsi were also detected from both Rhipicephalus ticks and Amblyomma ticks. In Homa Bay, Ehrichia ruminantium (17.5%) and Ehrichia canis (9.3%) were isolated from Amblyomma latum and Rhipicephalus pulchellus, as well as Anaplasma platys (14.4%) and Anaplasma ovis (14.4%) from Amblyomma and Rhipicephalus species. In determination of the occurrence of arboviruses among patients presenting with acute febrile illnesses, acute Bunyamwera 3 (0.9%) and Sindbis 2 (0.6%) infections were detected by RT-PCR and cell culture and Sindbis seroprevalence was determined by plaque assay. Though a significant proportion of these patients tested positive for low Plasmodium parasitemia, none were co-infected with Plasmodium parasites and arboviruses. This study highlights the presence and relative importance of zoonotic VBPs in both study areas.

No description available.

This thesis investigates the impact of international organizations on the making of international law by applying insights on how international organizations work—or fail to work—to the process of institutionalized treaty making. Specifically, I probe the relationship between the World Health Organization (“WHO”) and international infectious disease law, focusing in particular on the 2005 International Health Regulations (“2005 IHR”), which was negotiated, adopted, and is now being implemented under WHO’s auspices. The 2005 IHR is the most recent development in international infectious disease law, the history of which extends back to the beginning of international health cooperation in the mid-nineteenth century, before any international health organization was formed. Relying on secondary sources, WHO documents, archival materials, and personal interviews, I chronologically trace the evolution of international infectious disease law across changing institutional settings. I first examine the incremental growth of the older “barrier” approach to infectious disease regulation, initially developed in the absence of any international health organization and then with the aid of one of WHO’s predecessor organizations. I then analyze the decline of the barrier approach and the rise of the new “epidemiological” approach embodied by the 2005 IHR, with the aid of WHO. Based on my empirical analysis, I conclude that WHO has radically changed the process of making international infectious disease law as well as its content. On its own initiative and without member state demand, WHO’s permanent staff experimented with novel practices that subsequently became the basis for the 2005 IHR. WHO’s work reduced the length of formal negotiation needed to arrive at a new agreement and the uncertainty associated with adopting a novel regulatory system. Its influence also raises normative questions about the proper role of international organizations in making international law—questions that require further exploration.