INTRODUÇÃO: Toxocaríase é uma infecção parasitária de distribuição global, causada pela fase larval de Toxocara spp. Os hospedeiros naturais são cães e gatos, nos quais o parasita completa o ciclo chegando a fase adulta. Outros hospedeiros podem ser infectados pela fase larval do parasita, após ingestão de ovos embrionados do solo, mãos contaminadas, fomites, ou ingestão de carne ou vísceras de animais infectados. Em hospedeiros paratênicos o parasita não completa o ciclo, invadindo em estágio larval vísceras ou outros tecidos, onde podem sobreviver e induzir a patologia. O presente estudo teve como objetivo caracterizar o hamster (Mesocricetus auratus), como modelo experimental de toxocaríase, inicialmente através do estudo das lesões histopatológicas em fígado, pulmão e rim. A caracterização da resposta imunológica do modelo, foi feita através do estudo de citocinas envolvidas nas respostas Th1 e Th2, e foi sugerida uma correlação entre alterações glomerulares e depósitos de complexos antígenos-anticorpo pré-formados na circulação. MÉTODOS: Hamsters foram inoculados com ovos embrionados de Toxocara canis, e mantidos no biotério do Instituto de Medicina Tropical de São Paulo. O estudo histopatológico foi desenvolvido utilizando-se cortes parafinados corados por hematoxilina e eosina. Para detecção de antígenos nos tecidos foram realizadas reações imunohistoquímicas, utilizando-se anticorpo monoclonal e policlonal anti- Toxocara canis. Utilizando-se o soro dos animais infectados e animais controle, foi realizada pesquisa de antígeno e anticorpo por ELISA. Para pesquisa de imunoglobulinas IgG e IgM e complemento, foram utilizados cortes congelados de rins para realização de reação de Imunofluorescência. Fragmentos de rins foram incluídos para utilização em microscopia eletrônica, para detecção de antígenos de toxocara e de imune complexos. Para caracterização de resposta imunológica foram estudadas citocinas envolvidas na resposta Th1 e Th2 por técnica de RT-PCR. RESULTADOS: Os achados histopatológicos demonstraram desde o início da infecção, presença de larvas em maior número no fígado, seguido de pulmão e raramente rins. Em fígado remanescentes larvares foram visualizados cercados por reação inflamatória granulomatosa. Logo no início da infecção foi encontrado pneumonite intersticial e intraalveolar focal, e lesão renal com glomérulo apresentando hiperplasia focal de células mesangiais (glomerulite mesangio-proliferativa). Houve marcação de antígenos em todos os grupos de animais infectados, tanto pelo anticorpo monoclonal, como pelo policlonal. Depósitos de imunoglobulinas e complemento foram marcados em glomérulo por imunofluorescência A análise dos soros por ELISA, demonstrou na pesquisa de anticorpos aumento gradativo no decorrer da infecção, acompanhado de diminuição de antígenos. Depósitos de antígenos em glomérulos foram detectados por microscopia imonoeletrônica. No RT-PCR foi detectado aumento significativo do nível de IL-4, com tendência de elevação de IL-10 e IFN-?. CONCLUSÃO: O hamster demonstrou ser um modelo experimental eficiente para toxocaríase. Entretanto este modelo é mais adequado para infecções de curto prazo. A resposta imunológica avaliada por RT-PCR, com elevado nível da expressão de IL-4, sugere uma resposta Th2, mas a tendência de aumento de IL-10 e IFN-? poderia sinalizar uma resposta mista Th1 e Th2. Achados de depósitos de imunoglobulinas no glomérulo sugerem a possibilidade de que as manifestações renais com síndrome nefrótica em humanos possa vir a ter como base a toxocaríase / INTRODUCTION: Toxocariasis is a parasitic infection of global distribution, caused by the larval stage of Toxocara spp. The natural hosts are dogs and cats, in which the parasite completes the cycle reaching adulthood. Other hosts can be infected with the larval stage of the parasite, after ingestion of embryonated eggs from the soil, contaminated hands, fomites, or ingestion of meat or viscera of infected animals. In paratenics hosts the parasite not complete the cycle, encroaching on larval stage in viscera or other tissues where they can survive and induce pathology. The present study aimed to characterize the hamster, Mesocricetus auratus, as experimental model of toxocariasis, initially through the study of histopathological lesions in the liver, lung and kidney. The characterization of immune response model, was made through the study of cytokines Th1 and Th2 responses involved, and a correlation was suggested between glomerular changes and antibody-antigen complexes deposits preformed in the circulation. METHODS: Hamsters were inoculated with embryonated eggs of Toxocara canis, and kept in the bioterium of the Institute of Tropical Medicine of the São Paulo. The histopathologic study was developed using paraffin slides stained by hematoxylin and eosin. For detection of antigens in tissues immunohistochemistry reactions were performed using monoclonal and polyclonal anti-Toxocara canis sera. Using the serum of infected and control animals, search has been carried out of antigen and antibody by ELISA. For the search of immunoglobulins IgG, IgM and complement, were used slides prepared from frozen fragments of kidneys and a immunofluorescence reaction. Fragments of kidneys were included for electron microscopy to detect antigens of Toxocara and immune complexes. For characterization of Th1 and Th2 response cytokines involved were detected by RT-PCR technique. RESULTS: Histopathological findings demonstrated since the beginning of the infection the presence of larvae in greater numbers in the liver, followed by lung and rarely kidneys. In the liver larval remnants were surrounded by a granulomatous inflammatory reaction. Early in the infection was found interstitial pneumonitis with intraalveolar focal inflammatory infiltrate and renal injury with glomerulus showing mesangial cell focal hyperplasia (mesangioproliferative glomerulonephritis). There were the presence of antigens in all groups of animals infected detected by both the monoclonal and polyclonal antibodies. Deposits of immunoglobulin and complement were present in glomerulus by immunofluorescence analysis. ELISA, showed that the presence of antibodies increased gradually in the course of infection, accompanied by progressive diminution of antigens. Clusters of antigen/s were detected by immunoelectron microscopy. RT-PCR showed a significant increase of IL-4, with a tendency of increase of IL- 10 and IFN-?. CONCLUSION: The hamster has proved to be an efficient experimental model for toxocariasis. However this model is best suited for short-term infections. The immune response evaluated by RT-PCR, with high level of expression of IL-4, suggests a Th2 response, but the trend of increase of IL-10 and IFN-? might suggest a Th1 and Th2 mixed response. Findings of immunoglobulin deposits in glomeruli suggests the possibility that the renal manifestations with nephrotic syndrome in humans might have, in certain circunstances, as a basis the toxocariasis

The spotted fever group rickettsiae are emerging diseases. They cause damage in their hosts by invading the endothelium in small to medium-sized blood vessels, which results in vasculitis that can cause clinical manifestations from most organs. The present thesis describes the prevalence of Rickettsia helvetica in ticks, the incidence of rickettsial infection based on seroreactivity and seroconversion in humans and their symptoms, from different parts of Sweden and the Åland Islands in Finland. This was accomplished through serological analysis of both retrospective and prospective serum samples from confirmed and suspected tick-bitten individuals compared to individuals with no knowledge of tick exposure (blood donors). We found a comparable seroprevalence to Rickettsia spp. in different geographical areas where ticks are present; it was also comparable to the seroprevalence of Borrelia spp. Seroprevalence was also more common, as suspected, in the tick-exposed group compared to blood donors. In comparison with co-infections with other tick-borne infections (Anaplasma spp. and Borrelia spp.), we could conclude that co-infections do exist and that, based on clinical findings, it is difficult to distinguish which microorganism causes certain clinical manifestations. For reliable conclusions regarding the causative microorganism, the diagnosis should basically rely on diagnostic tests. In comparison with Borrelia spp., seroconversion to Rickettisa spp. was more common in the areas we investigated, indicating that rickettsiosis is a common tick-borne infection in Sweden and most likely underdiagnosed. When investigating patients with meningitis, we found R. felis in cerebrospinal fluid from two patients with subacute meningitis. This was the first report in which R. felis was found and diagnosed in patients in Sweden. The patients recovered without sequelae and without causal treatment. To provide guidelines on when to treat Rickettisa spp. infections, more investigations are needed. The present thesis shows that Rickettsia spp. are common in ticks and do infect humans. Rickettsial infection should be considered in both non-specific or specific symptoms after a tick bite. It was also shown in the thesis that flea-borne rickettsiosis (R. felis) occurs in Sweden and may cause invasive infections

In der vorliegenden Arbeit sollten die unterschiedlichen Manifestationsformen der Borreliose anhand verschiedener Parameter verglichen und der Verlauf einer Borrelioseerkrankung analysiert werden. Dabei konnte gezeigt werden, dass es sich um eine ganzjährige Erkrankung handelt, die jedoch - je nach Manifestationsform - verschiedene jahreszeitliche Gipfel aufweist. So kommen die Neuroborreliose und die Frühborreliose gehäuft im Frühling und im Sommer vor, während die Lyme Arthritis durchweg ganzjährig auftritt. Da die Borrelien erst nach ca. 12 Stunden Latenz auf den Wirt übertragen werden, sind es vor allem die unbemerkten Zeckenstiche, die eine Infektion verursachen. Deshalb ist es nicht verwunderlich, dass sich nur rund 55 % der Patienten an einen zurück liegenden Zeckenstich erinnern konnten. Bei der Untersuchung der verschiedenen Laborparameter ergaben sich keine wesentlichen Unterschiede zwischen den Manifestationsformen bezüglich Hämoglobingehalt, Blutsenkungsgeschwindigkeit, Thrombozyten und Leukozyten im Serum. Desweiteren ergaben sich keine Hinweise auf eine Beteiligung des Rheumafaktors oder antinukleärer Antikörper an den immunologischen Prozessen während einer Lyme Arthritis Erkrankung. IL-17 konnte mittels ELISA weder in Serum/Plasma noch in Synovialflüssigkeit signifikant erhöht nachgewiesen werden. Es bleibt jedoch unklar, ob dies aufgrund einer langen Lagerung der Proben der Fall war oder ob IL-17 tatsächlich nicht signifikant erhöht vorlag. Aus der Literatur kann man entnehmen, dass IL-17 bei den Abläufen einer Infektion mit B. burgdorferi eine wichtige Rolle zu spielen scheint. Dies könnte einen Ansatz für neue Behandlungsmethoden der Lyme Arthritis darstellen, weshalb die Durchführung weitere Untersuchungen wichtig ist. Durch die Analyse der initialen Serologie konnte die Antikörperreaktion während einer Infektion mit B. burgdorferi veranschaulicht werden, die mit der Bildung von IgM- Antikörpern beginnt und dann einen Shift zu IgG-Antikörpern vollzieht. Die Antigene p19, p31/34, p39 und p65 scheinen hierbei vor allem bei der Lyme Arthritis im Vordergrund zu stehen. Weiterhin konnte bei dieser Analyse beobachtet werden, dass teilweise zahlreiche serologische Untersuchungen bei ein und demselbem Patienten statt fanden, was darauf schließen lässt, dass diese Untersuchung zur Verlaufskontrolle benutzt wurde. Grundsätzlich ist die Serologie hierzu jedoch nicht geeignet. Vielmehr sollte sie lediglich zur Diagnosestellung dienen. In dieser Funktion ist sie bei korrekter Interpretation ein hervorragendenes diagnostisches Mittel. Leider werden serologische Untersuchungen jedoch häufig falsch interpretiert und angewendet, was dazu führt, dass Patienten häufiger oder auch länger als nötig antibiotisch behandelt werden. So erhielten auch rund 8 % der Lyme Arthritis Patienten mehr als 2 Zyklen antibiotischer Therapie. Die Nebenwirkungen dieser langwierigen antibiotischen Behandlung sind nicht zu vernachlässigen. Die aktuellen Leitlinien sehen im Gegensatz dazu nämlich nach dem zweiten antibiotischen Zyklus den Beginn einer Therapie mit DMARD ́s vor, selten kommen auch intraartikuläre Steroide zum Einsatz. Diese beobachtete „Überdiagnostik und Übertherapie“ der Borrelioseerkrankung spiegelt vermutlich die Angst vor chronischen Verlaufsformen in der Bevölkerung und teilweise auch unter Ärzten wider. Bereits in anderen Studien konnte die Existenz einer therapieresistenten Verlaufsform der Borreliose - insbesondere der Lyme Arthritis – nachgewiesen werden. In der vorliegenden Arbeit litten 24 % der Lyme Arthritis Patienten nach einer adäquaten antibiotischen Behandlung laut eigenen Angaben zum Zeitpunkt der Umfrage noch unter Gelenkbeschwerden. Ob es sich hierbei nun um eine schwerwiegendere Verlaufsform in Europa handelt, bleibt zunächst offen. Zur Objektivierung der Beschwerden, zum Ausschluss einer Zweitinfektion oder aber auch einer Fibromyalgie als mögliche Differentialdiagnose sind prospektive Studien mit klinischer Untersuchung der Patienten im Verlauf nötig. Eindeutig ist jedoch, dass durch eine frühzeitige antibiotische Behandlung nach Infektion das Fortschreiten der Erkrankung effektiv verhindert werden kann. Chronische Beschwerden gaben hingegen auch die Patienten mit zurückliegender Neuroborreliose an. Rund 20% der Patienten litten nach eigener Angabe unter Allgemeinsymptomen, die ihre Lebensqualität subjektiv wesentlich einschränkten. Der spezifische Zusammenhang zwischen diesen Beschwerden und einer Neuroborreliose bzw. einer Borrelioseerkrankung im Allgemeinen ließ sich jedoch nicht endgültig klären. Es wäre hierfür der Vergleich mit einer gesunden Kohorte und die Anwendung spezieller neuropsychologischer Untersuchungen nötig. / This paper should compare different manifestations of borreliosis based on various parameters as well as analyze the development of a borreliosis infection. It could be shown that it is a year-round disease reaching different seasonal peaks depending on its manifestation. As an example, neuro borreliosis and early borreliosis occur frequently in spring and summer, whereas lyme arthritis infections occur during the whole year. As borrelia are transmitted to the host only after 12 hours of latency, especially tick bites that go unnoticed cause infections. That is why it does not come as a surprise that only 55% of the patients were able to remember a tick bite in the past. Examining the different laboratory parameters, no substantial differences between the different types of manifestation regarding haemoglobin content, erythrocyte sedimentation rate, thrombocytes and leucocytes in the serum were discovered. In addition, there were no hints as to a contribution of the rheumatic factor or antinuclear antibodies to immunologic processes during a lyme arthritis infection. Using ELISA, IL-17 could neither be detected in serum/plasma nor in synovial liquid in significantly increased concentrations. However, it remains unclear whether this was due to the long storage of the samples or whether IL-17 was actually not significantly increased. According to literature on this topic, IL-17 seems to play a major role during the course of an infection with B. burgdorferi. This could be a starting point for new ways of treating lyme arthritis and makes further research necessary. By analyzing initial serology, the antibody reaction during an infection with B. burgdorferi could be shown. It starts with the creation of IgM antibodies and then shifts to IgG antibodies. The antigenes p19, p31/34, p39 and p65 seem to be especially important when it comes to lyme arthritis. The analysis also made it clear that in some cases a single patient was exposed to numerous serological examinations what makes it probable that this examination was used to control the course of the infection. Basically, serology is not a suitable instrument to do so. It should rather be used to diagnose. Interpreting it correctly, it is in that function an excellent means of diagnosing. Unfortunately, serological examinations are often interpreted and applied in the wrong way, what leads to a more frequent or unnecessarily long antibiotics treatment of patients. Around 8% of lyme arthritis patients received more than two cycles of antibiotics treatment. The side effects of this longsome antibiotics treatment are not to be neglected. In contrast, current guidelines recommend a DMARDs therapy after the second cycle of antibiotics treatment, in rare cases intraarticular steroids are used. This observed “overdiagnostics and overtherapy” of borreliosis infections presumably reflects the fear of chronic infections among the population and partly among doctors. There have already been other studies proving the existence of a form of borreliosis infection resistant to therapy – especially regarding lyme arthritis. According to this paper, 24% of lyme arthritis patients said, when they were interrogated, that they were still suffering from arthralgia after an adequate antibiotics treatment. It remains open whether this is just a more serious progression in Europe. To come to an objective judgement of the pains, to exclude a second infection or a fibromyalgia as a potential differential diagnosis, prospective studies invoving clinical examinations of the patients are necessary. However, it is beyond any doubt that an early antibiotics treatment after the infection can effectively prevent the progress of the infection. Nevertheless, also patients with an overcome neuroborreliosis reported chronic pains. About 20% of the patients reported that they were suffering from general syndromes limiting their quality of life subjectively essentially. The specific connection between these pains and a neuroborreliosis respectively a borreliosis infection in general could, however, not be proven in the end. For this, it would be necessary to make a comparison with a healthy cohort and to apply special neuropsychological studies.

Made available in DSpace on 2014-08-20T13:32:51Z (GMT). No. of bitstreams: 1 dissertacao_elizandra_schoenardie.pdf: 268602 bytes, checksum: 08408f44b3e993faf7cf221741aa401e (MD5) Previous issue date: 2005-06-24 / The Visceral Larva Migrans (VLM) is a zoonotic disease caused by the helminth Toxocara cannis. The precocious diagnosis of this disease in humans is very important to determinate the evolution of the clinical case and the patient's treatment. The goal of the first experiment was determinate the presence of antibodies anti-T. cannis in children from Pelotas through the Indirect Enzyme-Linked Immunosorbent Assay (ELISA) front to the antigen TES, as well as to define the pattern of bands recognized by the positive serums in ELISA through the "Western blotting . For this experiment 427 serums from children, with ages between one to eleven yeas old was tested, those serums was adsorved with AgSoAl and determinate that 50,6% was positive for antibodies anti-TES, showing a significant association among the positive children for antibodies anti-TES and the contact with dogs and cats. This association was also observed in the children's different age groups, but not regarding the gender of the same ones. To perform the Western Blotting , 70 serums witch give a positive result in the Indirect ELISA was been used, all serums recognize glicoproteic bands in the range between 30 and 120 kDa. Was observed a diminution in the crusade reaction with AgSoAl when the adsorved and not adsorved serums with this antigen has been test in the Western blotting , where a band of 30 kDa demonstrate to be an important glicoprotein to specific diagnosis of VLM. In the second experiment, 25 mice BALB/c were inoculated with approximately 1000 eggs containing the larvae L3. Each fifteen days blood collection was made through the reto orbital plexus until the 105 days after de animals infection. The serums was tested in the Indirect ELISA using the Antigen TES and urea 6M in order to discriminate recent and late infection, through the percentile of avidity of the IgG in the different days after the infection. A low percentile of avidity was observed to the 15 days after inoculation (between 7,25 and 27,5%). After 60 days of infection, all the animals presented avidity between 31,4 and 58%. This result suggests that in mice BALB/c, to the 60 days after infection the chronic phase of VLM is already established. / A Larva Migrans Visceral (LMV) é uma doença zoonótica que possui como principal agente etiológico o helminto Toxocara canis. O diagnóstico precoce da doença no homem é importante para estudos de evolução clínica e tratamento do paciente e os inquéritos epidemiológicos para determinar a freqüência da infecção em uma população. Por isso, o primeiro experimento teve como objetivo, determinar a presença de anticorpos anti-T. canis em crianças da região de Pelotas através de Enzyme linked immnosorbent assay (ELISA) com o antígeno de excreção e secreção de larvas de Toxocara canis (TES), bem como definir, através de Western blotting , o padrão de bandas do TES reconhecidas pelos soros positivos ao ELISA. Foram ensaiados no ELISA Indireto 427 soros de crianças de um a 12 anos de idade adsorvidos com antígeno somático de Ascaris lumbricoides e determinado que 50,6% apresentaram anticorpos anti-TES, ocorrendo uma associação significativa entre as crianças positivas e o contato com cães e gatos. Esta associação também foi observada em diferentes faixas etárias das crianças, mas não com relação ao sexo das mesmas. Setenta soros positivos no ELISA foram ensaiados no Western blotting e todos reconheceram frações proteicas entre 30 e 120 kDa. Uma diminuição da reação cruzada com o AgSoAl foi observada quando soros adsorvidos com este antígeno foram testados no Western blotting , sendo que uma fração antigênica de 30 kDa apresentou-se como uma proteína importante para o diagnóstico específico da LMV. No segundo experimento, 25 camundongos BALB/c foram inoculados com aproximadamente 1000 ovos contendo a larva infectante (L3). Colheitas quinzenais de sangue foram realizadas através do plexo retro orbital até os 105 dias pós-infecção dos animais. Os soros foram ensaiados no ELISA Indireto utilizando o antígeno TES e a uréia 6M a fim de discriminar infecção recente e tardia, através do percentual de avidez da IgG nos diferentes dias após a infecção. Um baixo percentual de avidez, característico da infecção aguda, foi observado aos 15 dias pós-inoculação (entre 7,3 e 27,5%). Após 60 dias de infecção, todos os animais apresentaram avidez entre 31,4 e 58%. Através destes resultados, sugere-se que em camundongos BALB/c, aos 60 dias pós-infecção a fase crônica da LMV já está estabelecida.

Die Wahl des Themas ist mit einem sehr einfachen Umstand verbunden: Die St. Petersburger Philharmonische Gesellschaft ist die älteste Musikorganisation Russlands. Am Beispiel dieser Gesellschaft kann man anschaulich die „Musica migrans“ thematisieren.

Tez (Tıpta Uzmanlık) - Süleyman Demirel Üniversitesi, Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, 2007. / Bibliyografya var.

The spotted fever rickettsia, Rickettsia helvetica, is an endemic tick-borne bacteria in Sweden. It causes infections in humans, manifested as aneruptive fever, headache, arthralgia and myalgia, and sometimes an inoculation eschar or a rash. There have also been two known cases of human infections with R. felis in Sweden. The present thesis starts by investigating dispersal of ticks and Rickettsia spp. by migrating birds flying from Africa to Europe. Almost 15,000 birds were searched and 734 ticks collected, mainly of the species Hyalomma marginatum complex. Almost half (48%) of the ticks were infected with Rickettsia spp., 96% of which was R. aeschlimannii, the remaining R. africae and undefined species. The next study focused on questing ticks over a large area in Sweden and determining the prevalence of Rickettsia spp., Anaplasma spp. and Coxiella burnetii. Rickettsia spp. was found in 9.5-9.6% of the ticks and A. phagocytophilum in 0.7%; no C. burnetii was found. The last three papers in the thesis focused on the clinical presentation of rickettsiosis, the symptoms associated with the infection in general and particularly in patients with neurological complications. A tick-exposed population in Sweden was investigated to gain a better understanding of symptoms due to rickettsioses, also in relation to co-infections with other tick-borne bacteria. Based on symptoms, it was not possible to distinguish what pathogen caused the infections. Most patients had erythema migrans, some had serological reactions to Rickettsia spp., Borrelia spp. or co-infections by Rickettsia spp., Borrelia spp. and/or Anaplasma spp. In the fourth and fifth papers, we found associations between antibodies against Rickettsia spp. and sudden deafness (in 10-24% of patients) and facial nerve paralysis (in 8.3-25% of patients). In three patients R. felis was detected in the cerebrospinal fluids.    Briefly, the thesis helps to clarify our knowledge about tick dispersal, shows a narrower prevalence estimate of Rickettsia spp. in Swedish ticks, and illuminates symptoms of rickettsioses and co-infections with other tick-borne infections. It also shows that presence of erythema migrans may be explained by more than Lyme disease and indicates a possible association between rickettsiosis and sudden deafness and facial nerve paralysis.