Objective: To study the presence of Bartonella bacilliformis in ticks collected from two wild mammals in Madre de Dios, Peru. Results: A total of 110 ticks were collected. Among the 43 Amblyomma spp. extracted from the 3 Tapirus terrestris only 3 were positive for B. bacilliformis. In addition, 12 out of the 67 Rhipicephalus (Boophilus) microplus obtained from the 3 Pecari tajacu were positive for B. bacilliformis. For the first time B. bacilliformis have been detected in arthropods other than Lutzomyia spp. Further studies are required to elucidate the possible role of ticks in the spread of South American Bartonellosis. / This work was supported by Cienciativa of CONCYTEC Peru, under the contract N° 164‑2016‑FONDECYT. Dr. Lescano is sponsored by the training grant D43 TW007393 awared by the Fogarty International Center of the US National Institutes of Health. / Revisión por pares / Revisión por pares

Background The lack of an effective diagnostic tool for Carrion’s disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches. Methodology/Principal Findings We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. Methodology/Principal Findings We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. Conclusions/Significance From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion’s disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques.

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2017-05-26T19:06:37Z No. of bitstreams: 2 Tese - Eduardo Sugizaki - 2011.pdf: 1614665 bytes, checksum: c1512eaa0135859f4c7c6fa48dfb2d02 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-05-30T10:20:42Z (GMT) No. of bitstreams: 2 Tese - Eduardo Sugizaki - 2011.pdf: 1614665 bytes, checksum: c1512eaa0135859f4c7c6fa48dfb2d02 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-05-30T10:20:42Z (GMT). No. of bitstreams: 2 Tese - Eduardo Sugizaki - 2011.pdf: 1614665 bytes, checksum: c1512eaa0135859f4c7c6fa48dfb2d02 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2011-12-05 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This doctoral thesis is a historical and epistemological research of the clinical age of Carrion's disease (1842-1885) and of the early age of his bacteriologic study (1885-1913). That´s a disease born within a historical narrative about the medical effects of experimental inoculation that the medical student Daniel Alcides Carrión, was made to do in August 1885. According to the peruvian historical narrative about the origins and consequences of the medical experiment, the student was inoculated with the blood of a disease known as Peruvian wart, but developed and died, in October five, from the disease called Oroya fever. The experiment would then have shown that there are not two diseases, but only one, that which, since then, bears his name as a tribute to the heroic gesture of inoculation. According to current knowledge about the disease, it has two stages in its evolution or syndromes. A febrile phase, anemiante, acute and of high risk for the patient's life; another phase, which follows the first, is the development of a characteristic skin neoformation, with the form of papules blood, called Peruvian wart. The historiography of the disease, after 1940, suggests that this duality of pathological manifestation represented the clinical difficulty explaining the genesis and outcome of the experiment of Carrion. However, research of documents relevant to the experiment of 1885, made in the first chapter of our thesis, showed that the experiment was not aimed at overcoming the dichotomy of two diseases. The student already had an advanced understanding of the overall clinical progression of the disease, including its two stages. From our investigation, moreover, we conclude that the peruvian medicine considered demonstrated that the inoculation produced Oroya fever, but the peruvian medicine does not present anywhere the ways in which one can recognize the Oroya fever. Due to these problems was undertaken in the second chapter, the study of the period of foundation of Peruvian wart clinic (1842-1872) and came to the conclusion that at this time there already was a complete understanding of the evolution of disease, including its stages and forms of development. At this point in our investigation, the existence of the notion of Oroya fever has appeared as unnecessary for the clinical knowledge of the disease. Therefore, the third chapter of our thesis investigated the period from 1872 to 1885, a time when the notion of Oroya fever appears in medical literature. Our investigation has shown that the notion appeared in Peruvian medicine because it regarded the disease as being a strictly benign, afebrile rash. Consequently, the Peruvian nosological dualism appears as a result of its inability to explain the phenomena of the general economy’s commitment that anticipate the eruption of Peruvian wart. With this new understanding of the clinical description of disease, we retake, in the fourth chapter, what happened in the study of disease, after the experiment of Carrión. A review of medical literature of the period between 1885 and 1898 showed that the historical narrative of Lima on the 7 experiment of Carrion became a guiding discipline of all research on the disease of this new period. During one stage of that Peruvian research (1885-1898), required to remain still only clinic, the study of disease was a prisoner of its assumptions - it became necessary, to keep the interpretation of the experiment as true, to define the concept of Oroya fever. But the final solution was not reached because the effort of maintaining doctrinal assumptions was placed above the results of observation. This closed circle began to be broken with the discovery of Tamayo, in 1905, that part of what physicians called Oroya fever was the result of a very common complication of severe cases of Peruvian wart with paratyphoid bacilli. The fourth chapter is concerned, yet, to the period after the discovery of Tamayo, from 1905 to 1913, to know the fate of Oroya fever. This research shows that the Peruvian medicine did not give up of the traditional historical narrative about the Carrion’s experiment. After Tamayo’s explanation of the reasons of part of what is called Oroya fever, the Peruvian medicine refused to give up of the use of this notion. Arce, the main Peruvian defender of the traditional historical narration about the disease, in 1913, meaning to seek the recovery of significance of Oroya fever that kept alive the dual polarity needed to remain true to the idea that, in 1885, has been demonstrated the unity between the two diseases. This doctoral thesis has obtained, thus, the construction of a new history of knowledge of Carrion’s disease, for the period 1842 to 1913. / Esta tese doutoral é uma investigação histórica e epistemológica da idade clínica da doença de Carrión (1842-1885) ao início da idade bacteriológica de seu estudo (1885-1913). Trata-se de uma doença nascida no interior de uma narrativa histórica médica sobre os efeitos da inoculação experimental que o estudante de medicina Daniel Alcides Carrión, se fez fazer em agosto de 1885. Segundo a narrativa médica limenha sobre o experimento, suas origens e suas consequências, o estudante inoculara-se com o sangue de uma doença conhecida como verruga peruana, mas desenvolveu e morreu, em cinco de outubro daquele mesmo ano, da doença chamada febre de Oroya. O experimento teria, assim, demonstrado que não há duas doenças, mas uma só que, desde então, leva seu nome, como homenagem ao gesto heroico da inoculação. Segundo o conhecimento atual sobre a doença, ela possui duas fases ou síndromes em sua evolução. Uma febril, anemiante, aguda e de alto risco para a vida do doente. Outra, que sucede a primeira, é o desenvolvimento de uma neoformação cutânea característica, com a forma de pápulas sanguíneas, chamada verruga peruana. A historiografia sobre a doença, posterior a 1940, sugere de que esta dualidade de manifestação patológica representou a dificuldade clínica que explica a gênese e o resultado do experimento de Carrión. Entretanto, a investigação dos documentos pertinentes ao experimento de 1885, feita no primeiro capítulo de nossa tese, mostrou que o experimento não visava a ultrapassar a dicotomia de duas doenças. O estudante já detinha uma compreensão clínica global avançada da evolução da doença, incluindo as suas duas etapas. Da nossa investigação, ademais, resultou que a medicina limenha deu por demonstrado que a inoculação produziu a febre de Oroya, mas ela não apresentou, em lugar algum, os modos pelos quais se pode reconhecer a febre de Oroya. Em função destes problemas, empreendeu-se, no capítulo segundo, o estudo do período de fundação da clínica da verruga peruana (1842 a 1872) e chegou-se à conclusão de que, nesta época, já havia uma compreensão global da evolução completa da doença, incluindo as suas etapas e formas de desenvolvimento. A esta altura da nossa investigação, a existência da noção de febre de Oroya já aparecia como desnecessária para o conhecimento clínico da doença. Por isso, o capítulo terceiro de nossa tese investigou o período de 1872 a 1885, época em que a noção de febre de Oroya aparece na literatura médica. Nossa investigação mostrou que a noção surgiu na medicina limenha porque esta concebia a doença como sendo estritamente uma erupção cutânea, apirética e benigna. Consequentemente, o dualismo nosológico peruano aparece como resultado da sua incapacidade de explicar os fenômenos de comprometimento geral da economia que antecipam a erupção da verruga peruana. Com essa nova compreensão da descrição clínica da doença, retoma-se, no quarto capítulo, o que aconteceu, no estudo da doença, depois do experimento de Carrión. A análise da literatura médica do período de 1885 a 1898, aí realizada, mostrou que a narrativa 5 histórica limenha sobre o experimento de Carrión tornou-se uma disciplina ordenadora de toda pesquisa sobre a doença deste novo período. Durante uma etapa da pesquisa (1885-1898), obrigada a ainda permanecer apenas clínica, o estudo da doença esteve prisioneiro de seus pressupostos. Tornou-se necessário, para manter como verdadeira a interpretação do experimento, definir o conceito de febre de Oroya. Mas não se chegava à solução definitiva porque o esforço de manutenção dos pressupostos doutrinários era colocado acima dos resultados da observação. Este círculo fechado começou a ser quebrado com a descoberta de Tamayo, em 1905, de que parte daquilo que os clínicos chamavam de febre de Oroya era resultado de uma complicação muito frequente dos casos graves de verruga peruana por bacilos paratifoides. O capítulo quarto ocupa-se, ainda, do período posterior à descoberta de Tamayo, de 1905 a 1913, para conhecer o destino da febre de Oroya. Desta investigação resulta que a medicina peruana não abriu mão da narrativa histórica tradicional do experimento de Carrión. Depois que Tamayo deu a explicação das razões de parte daquilo a que se chamara de febre de Oroya, a medicina peruana não aceitou desistir do uso desta noção. Arce, o principal defensor da narração histórica tradicional sobre a doença, em 1913, procurou recuperar uma significação para a febre de Oroya que mantivesse viva a polaridade dualista necessária para que permanecesse verdadeira a ideia de que, em 1885, foi demonstrada a unidade entre duas doenças. A presente tese doutoral obteve, assim, a construção de uma nova história do conhecimento da doença de Carrión, relativa ao período de 1842 a 1913.