Orientador: Lucia Helena O'Dwyer / Banca: Elizabeth Moreira dos Santos Schmidt / Banca: Gervásio Henrique Bechara / Resumo: Hepatozoon canis é um hemoparasita transmitido por carrapatos que afeta cães em diversas regiões do mundo. Rhipicephalus sanguineus é considerado seu vetor biológico, entretanto, é possível que existam outros vetores. No Brasil, H. canis é descrito principalmente em áreas rurais, onde cães podem ser infestados por outras espécies, incluindo Amblyomma spp. O objetivo do presente trabalho foi infectar experimentalmente diferentes espécies de carrapatos com H. canis, a partir de cães naturalmente infectados. Para isso, foram utilizadas ninfas de A. cajennense e R. sanguineus e adultos de A. cajennense e A. ovale. Após o ingurgitamento dos espécimes em animal infectado, foi realizada pesquisa de formas evolutivas do protozoário na hemolinfa, intestino e hemocele dos ixodídeos, por técnicas citológicas e histológicas. Alguns carrapatos foram submetidos ao diagnóstico molecular pela técnica de PCR. Somente em duas fêmeas de A. cajennense foram observadas estruturas morfologicamente semelhantes a Hepatozoon spp. e uma fêmea de A. ovale apresentou oocistos na hemocele. Além disso, duas fêmeas de A. ovale e uma de A. cajennense foram positivas na PCR. Entretanto, não é possível caracterizar essas espécies como vetores de H. canis, pois foram usados muitos exemplares nas infecções experimentais e poucos foram capazes de se infectar com o parasita. É provável que R. sanguineus tenha pouca importância epidemiológica na transmissão de H. canis no Brasil, pois este estudo, assim como pesquisas anteriores, demonstraram que a espécie foi incapaz de se infectar experimentalmente com o protozoário / Abstract: Hepatozoon canis is a hemoparasite that afflicts dogs in several regions of the world. Rhipicephalus sanguineus is considered to be the biologic vector of this agent, although other species may act as possible vectors. In Brazil, H. canis is described mainly in rural areas, where dogs are infested by other species, such as Amblyomma spp. The aim of this study was to transmit experimentally H. canis to different species of ticks, using naturally infected dogs. For this purpose A. cajennense and R. sanguineus nymphs and A. cajennense and A. ovale adults were used. After the repletion of specimens fed on a naturally infected animal, we searched for development forms of the protozoa in hemolymph, gut and hemocoel of the ticks by cytological and histopathologic examinations. Some ticks were also examined by molecular detection (PCR). Only two female ticks of the species A. cajennense contained structures morphologically similar to Hepatozoon spp. and oocysts were recovered from the hemocoel of an A. ovale female. In addition, two females of A. ovale and one female of A. ovale were positive through PCR. However, our results are not enough to characterize these species as vectors of H. canis mainly because many ticks were used in experimental infections and few were infected with the parasite. It is likely that R. sanguineus has little epidemiological importance in the transmission of H. canis in Brazil, since this research and previous researchers demonstrated that R. sanguineus was unable to experimentally get infected with the protozoan / Mestre

Submitted by Tatiana Oliveira (tsilva@icict.fiocruz.br) on 2012-06-05T18:40:11Z No. of bitstreams: 1 lucio_andre_v_dias_ioc_bp_0006_2010.pdf: 4820130 bytes, checksum: fcb218258dc3edcfe6059416e4b6d952 (MD5) / Made available in DSpace on 2012-06-05T18:40:11Z (GMT). No. of bitstreams: 1 lucio_andre_v_dias_ioc_bp_0006_2010.pdf: 4820130 bytes, checksum: fcb218258dc3edcfe6059416e4b6d952 (MD5) Previous issue date: 2010 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de janeiro, RJ, Brasil / Na presente tese, foram estudados aspectos da relação parasito-hospedeiro, constituída pelo protozoário parasito Hepatozoon caimani (Apicomplexa: Adeleorina) e seu hospedeiro vertebrado, o jacaré Caiman yacare (Crocodylia: Alligatoridae), na região do Pantanal Sul, Brasil. Com o objetivo de esclarecer os componentes epizootiológicos desta relação procuramos no primeiro momento examinar os níveis de prevalência e distribuição da parasitemia do H. caimani em uma população do C. yacare, procurando por fatores que poderiam estar associados ao maior risco de infecção, tais como gênero, tamanho/idade, e condição corporal, no período de julho de 2006 a fevereiro de 2008. Encontramos prevalência de 76% para toda a população amostrada (n=229) e o risco de infecção foi associado somente com o tamanho/idade dos jacarés, onde os animais filhotes não apresentaram infecção, os juvenis apresentaram 63%, adulto jovens e adultos foram 100% infectados. O H. caimani apresentou ainda uma distribuição agregada na população do C. yacare. De maneira a esclarecer os fatores responsáveis pela alta quantidade de jacarés infectados na região do Pantanal Sul, iniciamos estudos para investigar os possíveis fatores responsáveis por este fenômeno. Em um segundo momento, realizamos o levantamento dos culicídeos que realizam repasto sanguíneo nos C. yacare e verificamos infecções naturais pelo H. caimani somente em exemplares do gênero Culex, sobretudo no subgênero Melanoconion. Obtivemos infecções experimentais de C. yacare limpos após a ingestão de mosquitos Culex spp. com oocistos esporulados. A espécie incriminada como vetora primária do parasito foi Cx. (Mel.) theobaldi, devido a sua elevada abundância, preferência alimentar pelos jacarés e elevados níveis de infecção natural pelo H. caimani. Verificamos que alguns gêneros de anuros silvestres, e os próprios C. yacare, mediante o canibalismo, podem ser considerados hospedeiros intermediários do H. caimani, provavelmente contribuindo para a transmissão do mesmo no ambiente silvestre. Por fim, verificamos que gametócitos do H. caimani em jacarés C. yacare e C. latirostris, experimentalmente infectados, podem ser encontrados por períodos que variaram de três a seis meses nos crocodilianos, sugerindo que os jacarés podem servir como fonte de infecção para os vetores no campo por períodos relativamente longos / We studied aspects of the host-parasite relation beteween the protozoan parasite Hepatozoon caimani (Apicomplexa: Adeleorina) and its vertebrate host, the caiman Caiman yacare, in the Pantanal region, Brazil. We investigated the eTpizootiological components of this relation by surveying the prevalence and parasitemia of TH. caimani in a population of C. yacare and searched for the factors that could influence the infection rates, such as sex, size/age and Tbody condition from July 2006 to February 2008. We sampled 229 caimans and found a prevalence of 76% and the infection rate was related to caimans size/age. The young ones were not infected, the prevalence was 63% in the juveniles and 100% among the adult ones. THepatozoon caimani showed an aggregated pattern of distribuition. We also investigated the factors likely to have caused the high prevalence of H. caimani among caimans. By sampling the culicids that bite C. yacare, we found that the natural infection occurs only by Culex species, especially those belonging to the subgenus Melanoconion. Experimental infections of clean caimans C. yacare was retrieved after they ingested Culex spp. with sporulated oocysts. Culex (Mel.) theobaldi was pointed as the main vector because of its great abundance, preferance for biting caimans and high natural infection levels by H. caimani. Some wild anuran genera may also be pointed as paratenic hosts, as well as caimans C. yacare themselves through canibalism. Finally, H. caimani can be found in experimentally infected caimans C. yacare and C. latirostris for three up to six months, suggesting caimans as a perennial source of infection for the vectors in the environment

Orientador: Lucia Helena O'Dwyer de Oliveira / Resumo: Espécies do gênero Hepatozoon incluem um grupo diverso de parasitas com mais de 340 espécies descritas em anfíbios, répteis, aves e mamíferos. A hepatozoonose canina é uma hemoparasitose transmitida por carrapatos relatada em diversas regiões do mundo e pode estar relacionada às espécies Hepatozoon canis e Hepatozoon americanum. Em nosso país, diferentes estudos demonstraram que H. canis é a espécie envolvida na infecção dos cães e os relatos são comuns, principalmente, em áreas rurais. Trabalhos previamente conduzidos mostram que o vetor de H. canis em áreas rurais do Brasil permanece desconhecido, o que pode sugerir a existência de outros mecanismos de transmissão, como a predação de hospedeiros paratênicos. Assim, este trabalho teve como objetivo investigar a presença de Hepatozoon spp. em pequenos roedores silvestres capturados em fragmentos florestais nos arredores de propriedades rurais do município de Botucatu, São Paulo, Brasil, onde o parasita foi também estudado em cães domésticos. Foram analisadas 158 amostras sanguíneas de cães, pela reação em cadeia da polimerase (PCR) e pela técnica de extensão de sangue periférico. Amostras de sangue e tecidos de 67 roedores foram coletadas para pesquisa do parasita pela técnica de extensão sanguínea, histopatologia e também por diagnóstico molecular. Os resultados do presente estudo mostraram uma prevalência elevada de H. canis em cães de áreas rurais do município de Botucatu (66,45%). Além disso, as espécies de roedores silve... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Hepatozoon spp. belong to a diverse group of parasites with more than 340 species described in amphibians, reptiles, birds and mammals. Canine hepatozoonosis is a tick-borne disease reported throughout the world and can be related to Hepatozoon canis and Hepatozoon americanum. In our country, previously studies have shown that H. canis is the species involved in the infection of dogs and reports are common, especially in rural areas. Hepatozoon canis vectors in rural areas of Brazil remain unknown, which might suggest the existence of other transmission mechanisms, such as predation of paratenic hosts. Thus, this study aimed to investigate the presence of Hepatozoon spp. in wild rodents trapped in forest fragments surrounding rural areas of Botucatu, São Paulo, Brazil, where the prevalence of the parasite was also studied in domestic dogs. In dogs, the study was conducted in 158 blood samples using polymerase chain reaction (PCR) and peripheral blood extension technique. Blood samples and tissues from 67 rodents were collected for parasite blood extension technique, histopathology and for molecular diagnosis. The results showed a high prevalence of H. canis in dogs from rural areas of Botucatu (66.45%). In addition, species of Brazilian wild rodents are infected with Hepatozoon spp., other than H. canis. Additionally, the finding of monozoics cysts in the examined rodents may indicate that, in addition to intermediate hosts, small wild rodents may also act as paratenic hosts ... (Complete abstract click electronic access below) / Doutor

Die vorliegende epidemiologische Studie umfaßte 153 Hunde aus der Nähe von Athen, Griechenland. Um die Prävalenz arthropogener Infektionen abschätzen zu können, wurden Serumproben auf Antikörper gegen Leishmania infantum, Ehrlichia canis und Borrelia burgdorferi sowie auf Antigene von Dirofilaria immitis überprüft. Blutausstriche wurden auf das Vorkommen von Babesia canis und Hepatozoon canis untersucht. Außerdem wurden von den Hunden abgesammelte Zecken bestimmt. Bei 126 Hunden erfolgte eine klinische Allgemeinuntersuchung. Des weiteren wurden die serologischen Testverfahren ELISA und IFAT für den Nachweis von Antikörpern gegen Borrelia burgdorferi miteinander verglichen. Insgesamt waren 94 (61,4 %) der 153 untersuchten Hunde infiziert, 63 (41,2 %) Hunde wiesen Antikörper gegen Ehrlichia canis auf. Infektionen mit Borrelia burgdorferi wurden im ELISA insgesamt bei 43 (28,1 %), im IFAT bei 35 (22,9 %) Tieren nachgewiesen. Außerdem konnte bei 28 (18,3 %) Tieren eine Infektion mit Leishmania infantum und bei 20 (13,1 %) mit Dirofilaria immitis gefunden werden. Ein Nachweis von Babesia canis im Blutausstrich gelang bei vier (2,6 %) Hunden, von Hepatozoon canis nur bei einem (0,7 %) Hund. Für keinen der untersuchten Infektionserreger konnten signifikante Alters-, Geschlechts- oder Rasseprädispositionen festgestellt werden. Die Infektionsraten mit Leishmanien, Babesien, Dirofilarien und Borrelien waren bei den im Tierheim lebenden Hunden geringer als bei den Streunern. Vierundvierzig (28,8 %) der 153 getesteten Hunde waren gleichzeitig mit zwei, drei oder vier durch Arthropoden übertragene Erregerarten infiziert. Zweifachinfektionen kamen bei 29 (19,0 %), Dreifachinfektionen bei sieben (4,6 %) und Vierfachinfektionen bei acht (5,2 %) Tieren vor. In der Regel war das Risiko für Hunde, einen Erreger zu beherbergen, höher, wenn bereits eine Infektion mit einem anderen Erreger vorhanden war. Insgesamt waren 28 (18,3 %) Streuner und 15 (9,8 %) Tierheimhunde von Mehrfachinfektionen betroffen. Von den 94 mit mindestens einem der berücksichtigten Erreger infizierten Hunden wurden 75 (79,8 %) einer klinischen Untersuchung unterzogen. Dreiunddreißig (44,0 %) dieser Tiere zeigten Krankheitserscheinungen. Bei 21 (67,7 %) der 31 klinisch untersuchten und mit mehreren Erregern gleichzeitig infizierten Hunde konnten Krankheitssymptome diagnostiziert werden. Von 44 klinisch untersuchten Tieren, die nur mit einem Erreger infiziert waren, zeigten 12 (27,3 %) klinische Symptome. Die diagnostizierten Symptome waren bei allen untersuchten Erregern von einer starken Variabilität geprägt. Die Übereinstimmung der Testsysteme IFAT und ELISA für den Nachweis von Antikörpern gegen Borrelia burgdorferi lag bei 92,2 %. Die Testverfahren sind damit als nahezu gleichwertig anzusehen, wobei der ELISA etwas sensitiver zu sein scheint. Eine Infektionsgefahr besteht demnach in Griechenland vor allem für Ehrlichia canis, Borrelia burgdorferi und Leishmania infantum, aber auch für Infektionen mit Dirofilaria immitis besteht ein Risiko. Infektionen mit Babesia canis kommen in dieser Region seltener vor, es muss jedoch insbesondere bei geschwächten oder in Deutschland geborenen und somit hochempfänglichen Tieren mit Erkrankungen gerechnet werden. Hepatozoon canis ist nach den vorliegenden Ergebnissen in der Umgebung von Athen von geringer Verbreitung. Ein Rückschluß vom klinischen Bild auf das Vorliegen einer Infektion mit einem bestimmten Erreger kann aufgrund der Variabilität der Symptome in Verbindung mit häufig auftretenden Mehrfachinfektionen nicht gezogen werden. Die Möglichkeit von gleichzeitig vorliegenden Infektionen sollte in der Diagnostik und Therapie unbedingt berücksichtigt werden. Gute Haltungsbedingungen und eine tierärztliche Überwachung und Prophylaxe reduzieren offensichtlich die Inzidenz von Infektionen mit arthropodenübertragenen Erregern. Die überwiegende Anzahl (449 von 457 Exemplaren) der bestimmten Zecken gehörte der Art Rhipicephalus sanguineus an. Es wurden sechs Nymphen und 443 adulte Tiere gefunden. Die adulten Tiere verteilten sich auf 243 männliche und 200 weibliche Zecken. Bei acht Zecken handelte es sich um weibliche Ixodes ricinus. Bei aus Griechenland stammenden Hunden wie auch bei Tieren, die sich reisebegleitend dort aufgehalten haben, muss mit einer starken Infestation mit Rhipicephalus sanguineus gerechnet werden. Da Rhipicephalus sanguineus der Vektor für Ehrlichia canis, Babesia canis vogeli und Hepatozoon canis ist, sollten Prophylaxemaßnahmen eine geeignete Zeckenbekämpfung einbeziehen. / 153 dogs from the environs of Athens, Greece, were surveyed for tick infestation and arthropod borne infections. Serology was performed for Leishmania infantum, Dirofilaria immitis, Ehrlichia canis and Borrelia burgdorferi and bloodfilms were microscopically examined for Babesia canis and Hepatozoon canis. Ticks collected from the dogs were differentiated. 126 dogs underwent clinical examination. Suitability of an indirect immunofluorescent antibody assay (IFAT) and an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of antibodies to Borrelia burgdorferi was compared. Altogether 94 (61.4 %) dogs were infected with an arthropod borne pathogen, 63 (41.2 %) produced antibodies to Ehrlicha canis. ELISA detected Borrelia burgdorferi infection in 43 (28.1 %) dogs while IFAT was positive in 35 (22.9 %). 28 (18.3 %) dogs were infected with Leishmania infantum and 20 (13.1 %) with Dirofilaria immitis. Babesia canis was found in blood smears of four (2.6 %) dogs, Hepatozoon canis was detected only in one case (0,7 %). No association was found between the breed, age or sex of the dogs and any of the tested pathogens. The rate of infection with Leishmania, Babesia, Dirofilaria or Borrelia was lower in dogs living in the animal shelter than in those living as strays. 44 (28.8 %) of the 153 dogs examined were infected concurrently with two (n = 29, 19,0 %), three (n = 7, 4,6 %) or four (n = 8, 5,2 %) arthropod borne pathogens. In general the risk of infection was higher in dogs that were already infected with another pathogen. Altogether 28 (18.3 %) of the strays and 15 (9.8 %) of the dogs living in the animal shelter had a multiple infection. 75 of the 94 dogs infected with at least one of the tested pathogens were examined clinically. 33 (44.0 %) showed clinical symptoms. In 21 (67,7 %) of the 31 clinically examined dogs with multiple infection symptoms of disease were diagnosed. Twelve (27.3 %) of the 44 clinically examined dogs that were only infected with one pathogen had symptoms of some kind. Clinical symptoms varied considerably irrespective of the causative agent. The concordance of the IFAT and the ELISA for the diagnosis of Borrelia burgdorferi was 92.2 %. The serologic diagnosis of Borrelia burgdorferi with IFAT or ELISA can be regarded similarly suitable although ELISA seems to be somewhat more sensitive. In conclusion the risk of infection in Greece is particularly high for Ehrlichia canis, Borrelia burgdorferi and Leishmania infantum and Dirofilaria immitis as well. Babesia canis is obviously transmitted less. However dogs in poor condition or born in non endemic areas may acquire babesiosis in Greece. According to the results of this study Hepatozoon canis seems to be of negligible relevance in Greece. Due to the variability of the clinical symptoms and the frequent occurrence of multiple infections diagnosis based on the clinical picture only is not possible. Multiple infections should also be considered with respect to treatment. Infections with arthropod borne pathogens seem to occur less often in prophylactically treated dogs living under good conditions. Most of the examined ticks were identified as Rhipicephalus sanguineus (449 out of 457 specimens). Six nymphs and 419 adults were found. Of the adult Rhipicephalus sanguineus ticks 243 were males and 200 were females. Eight of the ticks were female Ixodes ricinus. In dogs originating from Greece as well as in dogs having accompanied tourists into Greece infestation with Rhipicephalus sanguineus is probable. As Rhipicephalus sanguineus is known to transmit Ehrlicha canis, Babesia canis vogeli and Hepatozoon canis prophylactic measures should include a suitable tick-control.

Die vorliegende epidemiologische Studie umfaßte 153 Hunde aus der Nähe von Athen, Griechenland. Um die Prävalenz arthropogener Infektionen abschätzen zu können, wurden Serumproben auf Antikörper gegen Leishmania infantum, Ehrlichia canis und Borrelia burgdorferi sowie auf Antigene von Dirofilaria immitis überprüft. Blutausstriche wurden auf das Vorkommen von Babesia canis und Hepatozoon canis untersucht. Außerdem wurden von den Hunden abgesammelte Zecken bestimmt. Bei 126 Hunden erfolgte eine klinische Allgemeinuntersuchung. Des weiteren wurden die serologischen Testverfahren ELISA und IFAT für den Nachweis von Antikörpern gegen Borrelia burgdorferi miteinander verglichen. Insgesamt waren 94 (61,4 %) der 153 untersuchten Hunde infiziert, 63 (41,2 %) Hunde wiesen Antikörper gegen Ehrlichia canis auf. Infektionen mit Borrelia burgdorferi wurden im ELISA insgesamt bei 43 (28,1 %), im IFAT bei 35 (22,9 %) Tieren nachgewiesen. Außerdem konnte bei 28 (18,3 %) Tieren eine Infektion mit Leishmania infantum und bei 20 (13,1 %) mit Dirofilaria immitis gefunden werden. Ein Nachweis von Babesia canis im Blutausstrich gelang bei vier (2,6 %) Hunden, von Hepatozoon canis nur bei einem (0,7 %) Hund. Für keinen der untersuchten Infektionserreger konnten signifikante Alters-, Geschlechts- oder Rasseprädispositionen festgestellt werden. Die Infektionsraten mit Leishmanien, Babesien, Dirofilarien und Borrelien waren bei den im Tierheim lebenden Hunden geringer als bei den Streunern. Vierundvierzig (28,8 %) der 153 getesteten Hunde waren gleichzeitig mit zwei, drei oder vier durch Arthropoden übertragene Erregerarten infiziert. Zweifachinfektionen kamen bei 29 (19,0 %), Dreifachinfektionen bei sieben (4,6 %) und Vierfachinfektionen bei acht (5,2 %) Tieren vor. In der Regel war das Risiko für Hunde, einen Erreger zu beherbergen, höher, wenn bereits eine Infektion mit einem anderen Erreger vorhanden war. Insgesamt waren 28 (18,3 %) Streuner und 15 (9,8 %) Tierheimhunde von Mehrfachinfektionen betroffen. Von den 94 mit mindestens einem der berücksichtigten Erreger infizierten Hunden wurden 75 (79,8 %) einer klinischen Untersuchung unterzogen. Dreiunddreißig (44,0 %) dieser Tiere zeigten Krankheitserscheinungen. Bei 21 (67,7 %) der 31 klinisch untersuchten und mit mehreren Erregern gleichzeitig infizierten Hunde konnten Krankheitssymptome diagnostiziert werden. Von 44 klinisch untersuchten Tieren, die nur mit einem Erreger infiziert waren, zeigten 12 (27,3 %) klinische Symptome. Die diagnostizierten Symptome waren bei allen untersuchten Erregern von einer starken Variabilität geprägt. Die Übereinstimmung der Testsysteme IFAT und ELISA für den Nachweis von Antikörpern gegen Borrelia burgdorferi lag bei 92,2 %. Die Testverfahren sind damit als nahezu gleichwertig anzusehen, wobei der ELISA etwas sensitiver zu sein scheint. Eine Infektionsgefahr besteht demnach in Griechenland vor allem für Ehrlichia canis, Borrelia burgdorferi und Leishmania infantum, aber auch für Infektionen mit Dirofilaria immitis besteht ein Risiko. Infektionen mit Babesia canis kommen in dieser Region seltener vor, es muss jedoch insbesondere bei geschwächten oder in Deutschland geborenen und somit hochempfänglichen Tieren mit Erkrankungen gerechnet werden. Hepatozoon canis ist nach den vorliegenden Ergebnissen in der Umgebung von Athen von geringer Verbreitung. Ein Rückschluß vom klinischen Bild auf das Vorliegen einer Infektion mit einem bestimmten Erreger kann aufgrund der Variabilität der Symptome in Verbindung mit häufig auftretenden Mehrfachinfektionen nicht gezogen werden. Die Möglichkeit von gleichzeitig vorliegenden Infektionen sollte in der Diagnostik und Therapie unbedingt berücksichtigt werden. Gute Haltungsbedingungen und eine tierärztliche Überwachung und Prophylaxe reduzieren offensichtlich die Inzidenz von Infektionen mit arthropodenübertragenen Erregern. Die überwiegende Anzahl (449 von 457 Exemplaren) der bestimmten Zecken gehörte der Art Rhipicephalus sanguineus an. Es wurden sechs Nymphen und 443 adulte Tiere gefunden. Die adulten Tiere verteilten sich auf 243 männliche und 200 weibliche Zecken. Bei acht Zecken handelte es sich um weibliche Ixodes ricinus. Bei aus Griechenland stammenden Hunden wie auch bei Tieren, die sich reisebegleitend dort aufgehalten haben, muss mit einer starken Infestation mit Rhipicephalus sanguineus gerechnet werden. Da Rhipicephalus sanguineus der Vektor für Ehrlichia canis, Babesia canis vogeli und Hepatozoon canis ist, sollten Prophylaxemaßnahmen eine geeignete Zeckenbekämpfung einbeziehen. / 153 dogs from the environs of Athens, Greece, were surveyed for tick infestation and arthropod borne infections. Serology was performed for Leishmania infantum, Dirofilaria immitis, Ehrlichia canis and Borrelia burgdorferi and bloodfilms were microscopically examined for Babesia canis and Hepatozoon canis. Ticks collected from the dogs were differentiated. 126 dogs underwent clinical examination. Suitability of an indirect immunofluorescent antibody assay (IFAT) and an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of antibodies to Borrelia burgdorferi was compared. Altogether 94 (61.4 %) dogs were infected with an arthropod borne pathogen, 63 (41.2 %) produced antibodies to Ehrlicha canis. ELISA detected Borrelia burgdorferi infection in 43 (28.1 %) dogs while IFAT was positive in 35 (22.9 %). 28 (18.3 %) dogs were infected with Leishmania infantum and 20 (13.1 %) with Dirofilaria immitis. Babesia canis was found in blood smears of four (2.6 %) dogs, Hepatozoon canis was detected only in one case (0,7 %). No association was found between the breed, age or sex of the dogs and any of the tested pathogens. The rate of infection with Leishmania, Babesia, Dirofilaria or Borrelia was lower in dogs living in the animal shelter than in those living as strays. 44 (28.8 %) of the 153 dogs examined were infected concurrently with two (n = 29, 19,0 %), three (n = 7, 4,6 %) or four (n = 8, 5,2 %) arthropod borne pathogens. In general the risk of infection was higher in dogs that were already infected with another pathogen. Altogether 28 (18.3 %) of the strays and 15 (9.8 %) of the dogs living in the animal shelter had a multiple infection. 75 of the 94 dogs infected with at least one of the tested pathogens were examined clinically. 33 (44.0 %) showed clinical symptoms. In 21 (67,7 %) of the 31 clinically examined dogs with multiple infection symptoms of disease were diagnosed. Twelve (27.3 %) of the 44 clinically examined dogs that were only infected with one pathogen had symptoms of some kind. Clinical symptoms varied considerably irrespective of the causative agent. The concordance of the IFAT and the ELISA for the diagnosis of Borrelia burgdorferi was 92.2 %. The serologic diagnosis of Borrelia burgdorferi with IFAT or ELISA can be regarded similarly suitable although ELISA seems to be somewhat more sensitive. In conclusion the risk of infection in Greece is particularly high for Ehrlichia canis, Borrelia burgdorferi and Leishmania infantum and Dirofilaria immitis as well. Babesia canis is obviously transmitted less. However dogs in poor condition or born in non endemic areas may acquire babesiosis in Greece. According to the results of this study Hepatozoon canis seems to be of negligible relevance in Greece. Due to the variability of the clinical symptoms and the frequent occurrence of multiple infections diagnosis based on the clinical picture only is not possible. Multiple infections should also be considered with respect to treatment. Infections with arthropod borne pathogens seem to occur less often in prophylactically treated dogs living under good conditions. Most of the examined ticks were identified as Rhipicephalus sanguineus (449 out of 457 specimens). Six nymphs and 419 adults were found. Of the adult Rhipicephalus sanguineus ticks 243 were males and 200 were females. Eight of the ticks were female Ixodes ricinus. In dogs originating from Greece as well as in dogs having accompanied tourists into Greece infestation with Rhipicephalus sanguineus is probable. As Rhipicephalus sanguineus is known to transmit Ehrlicha canis, Babesia canis vogeli and Hepatozoon canis prophylactic measures should include a suitable tick-control.

Little is understood about off-host behavior and ecology of immature Amblyomma maculatum Koch (Gulf Coast tick). A more complete understanding of this tick is essential to protect human and animal health. My research focused on seasonality and distribution of immatures in Mississippi, potential suitability of some insect and human hosts to larvae, and aspects of nymphal questing behavior. A single larva was collected (third off-host collection reported) when sampling A. maculatum habitat using a novel device. Collection of this larva in November expands the stage’s known seasonality and confirmed a prediction concerning seasonality of larval A. maculatum. Low frequency of immatures (8.3%) confirmed that they’re incredibly difficult to collect off-host. Nymphal collections peaked in March, and known seasonality was extended for both nymphs and adults. I examined known records, elucidating seasonality and distribution of A. maculatum in Mississippi. Either multiple generations per year or diapause are responsible for observed bi-modal distribution of immature collections. Additionally, I compiled the most extensive host record of immature A. maculatum in Mississippi and investigated seasonality patterns using USDA plant hardiness zones. I compiled the most complete record of ticks found on arthropods. Amblyomma americanum and A. maculatum were both confirmed to crawl onto arthropods, giving support to occasional, unintentional dispersal by phoresy. There was no conclusive evidence that larval A. maculatum feed on arthropods, however data supported feeding by larval A. americanum. These results have interesting implications regarding evolution of pathogens/endosymbionts. I provided the first evidence that larval A. maculatum can attach to humans. Rickettsia parkeri, a human pathogen transmitted by this species has recently been shown to be capable of transovarial transmission. Therefore, larval A. maculatum may provide another avenue of transmission. I have demonstrated that A. maculatum are difficult to collect off-host in part because they prefer to quest low to the ground. In choice studies, 5-cm-tall stems were most likely to be occupied by nymphs released into an array of stems. Low vapor pressure deficit encouraged questing, while higher VPD and warmer temperature increased questing height. These results may have implications in understanding host-seeking behavior in other tick species as well.