ASTMH 64th Annual Meeting. October 25-29, 2015 Philadelphia Marriott Downtown Philadelphia, Pennsylvania USA / Bartonella bacilliformis is the aetiological agent of Carrion's disease, an overlooked illness with a lethal febrile stage and a benign warty phase. Its endemic in Andean areas, mainly affecting Peru, but also reported in Ecuador, Colombia, Bolivia and Chile.

ASTMH 64th Annual Meeting. October 25-29, 2015 Philadelphia Marriott Downtown Philadelphia, Pennsylvania USA / The bacteria Bartonella bacilliformis is the etiological agent of Carrion’s disease, which is a neglected poverty-related disease, affecting Mountain Andean valleys of Peru, Colombia and Ecuador. This disease, in absence of treatment presents a high mortality during the acute phase, called Oroya’s Fever. The second phase is characterized by the development of dermal eruptions, known as “Peruvian wart”. This bacterium is a fastidious slow growing microorganism, being difficult and cumbersome to culture and isolate from clinical sources. Then, the available data about phylogenetic relationship in clinical samples are really scarce, but suggesting high variability. The aim of the study was to perform direct blood analysis of B. bacilliformis Multi Locus Sequence Typing (MLST), a genotyping tool, in patients with Oroya fever during an outbreak. The present study demonstrate that the direct blood MLST PCR is a technique useful in the phylogenic characterization of this fastidious microorganism endemic from Andean regions. In this study, we demonstrate that the outbreak of Oroya’s fever was caused by closely related Sequence Typing (ST) microorganisms and, additionally, new STs have been described.

juana.delvalle@upc.edu.pe / Objetivo General: Caracterizar a nivel epidemiológico molecular cepas de Bartonella bacilliformis en las diferentes fases de la infección (Fiebre, Verruga, Bacteriemia asintomática). Objetivos específicos: 1.- Describir la presentación clínica e inmunológica de la enfermedad causada por Bartonella bacilliformis en las diferentes fases de infección: Determinar la etiología y caracterizar la presentación fenotípica y epidemiológica. Caracterizar los parámetros bioquímicos y hematológicos. Cuantificar los niveles de anticuerpos específicos para Bartonella bacilliformis

Bartonella bacilliformis is a facultative, intracellular, aerobic, Gram-negative coccobacillus causing the so-called Carrión's disease, a human infection endemic to specific areas mainly inhabited by low-income communities of Peru but also present in other Andean communities. It is considered a truly neglected tropical disease and is transmitted through the bite of female sandflies of the genus Lutzomyia [1]. Carrión's disease has two different clinical presentations; an initial febrile and haemolytic anaemia phase, known as Oroya fever, which has a mortality rate ranging from 44% to 88% in untreated patients; and a second phase characterised by the development of dermal eruptions known as Peruvian wart [1,2]. / The study was supported by internal funds from the Universidad Peruana de Ciencias Aplicadas (Lima, Peru); by a grant from the Instituto de Salud Carlos III, Spain [PI11/00983], which included FEDER funds; by the UBS Optimus Foundation; and by Generalitat de Catalunya, Departament d’Universitats, Recerca i Societat de la Informació [2014 SGR 26]. MJP has received a postdoctoral fellowship from CONCYTEC/FONDECYT. JR has received a fellowship from the program I3 of the ISCIII [grant no. CES11/012]. / Revisión por pares

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

Esta tesis se encuentra en proceso de registro como patente. / Antecedentes: La Enfermedad de Carrión es una patología importante que requiere un diagnóstico precoz para su manejo a fin de disminuir la morbimortalidad de la misma. Objetivo: Desarrollar una prueba inmunocromatográfica para la detección rápida de Bartonella bacilliformis usando anticuerpos policlonales de conejo etiquetados con oro coloidal. Metodología: Los anticuerpos policlonales contra Bartonella bacilliformis fueron obtenidos mediante la inmunización de conejos con la proteína GroEL de Bartonella bacilliformis. Se utilizó oro coloidal de 40 nm para la conjugación el anticuerpo primario obtenido. La sensibilidad analítica se evaluó con diferentes concentraciones de B. bacilliformis incluidas en un rango de 1x101 UFC/mL a 1x105 UFC/mL. La especificidad analítica o reacción cruzada se analizó con microorganismo de diferentes especies, incluyendo Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus viridans, Chlamydophila pneumoniae, Escherichia coli y Candida spp. Resultados: Las concentraciones óptimas del anticuerpo de captura y el anticuerpo de revestimiento (línea T) fueron de 2 mg/mL y 0.2 mg/mL respectivamente. Se determinó que la sensibilidad analítica de la prueba se encuentra en un rango de 1x102 UFC/mL y 1x101 UFC/mL. No se evidenciaron reacciones cruzadas con los microorganismos evaluados. Se determinó que el tiempo de corrida óptimo para el registro de resultados es de 20 minutos y el volumen mínimo requerido de la muestra analizada fue de 50 µL. Conclusión: Se desarrolló la primera prueba inmunocromatográfica para la detección rápida, sensible y específica de Bartonella bacilliformis utilizando anticuerpos policlonales contra la proteína GroEL, la cual deberá ser validada mediante estudios posteriores. / Background: Carrión's disease is an important disease that requires a timely diagnosis and management to reduce its morbimortality. Objective: Develop a lateral flow assay for the rapid detection of Bartonella bacilliformis using colloidal gold-labeled rabbit polyclonal antibodies. Materials and methods: Polyclonal antibodies against Bartonella bacilliformis were produced by the immunization of rabbits with GroEL protein purified from Bartonella bacilliformis. Colloidal gold of 40 nm was used for the conjugation process with the rabbit polyclonal antibodies. The analytical sensitivity was evaluated by testing solutions of B. bacilliformis with different concentrations ranging between 1 x 101 to 1 x 105 CFU/mL. Analytical specificity was determined by testing cross-reactivity with microorganisms from different species, including Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus viridans, Chlamydophila pneumoniae, Escherichia coli and Candida spp. Results: The optimal concentrations for the capture antibody and the coating antibody (T-line) were 2 mg / mL and 0.4 mg / mL, respectively. The analytical sensitivity was determined to be between 1x102 CFU/mL and 1x101 CFU/mL. There were no cross-reactions observed with the groups of bacteria used in this study. We determined that the flowing time and volume required for an optimum signal to be generated was 20 minutes and 50 µL, respectively. Conclusions: We developed the first gold-based lateral flow assay for the rapid, sensitive and specific detection of Bartonella bacilliformis using polyclonal antibodies against the protein GroEL, which needs to be validated in future studies. / Tesis

Infective endocarditis due to Bartonella bacilliformis is rare. A 64-year-old woman, without previous heart disease, presented with 6 weeks of fever, myalgias, and arthralgias. A systolic murmur was heard on the tricuspid area upon examination, and an echocardiogram showed endocardial lesions in the right atrium. Bartonella bacilliformis was isolated in blood cultures, defining the diagnosis of infective endocarditis using Duke’s criteria. Subsequently, the patient developed clinical and laboratory features compatible with antineutrophil cytoplasmic antibody-associated vasculitis. This case presents an uncommon complication of B. bacilliformis infection associated with the development of systemic vasculitis.

Objective This study investigated an outbreak of Bartonellosis in a coastal region in Peru. Results A total of 70 (n = 70) samples with clinical criteria for the acute phase of Bartonellosis and a positive peripheral blood smear were included. 22.85% (n = 16) cases of the samples were positive for Bartonella bacilliformis by PCR and automatic sequencing. Of those positive samples, 62.5% (n = 10) cases were positive only for B. bacilliformis and 37.5% (n = 6) cases were positive to both Mycobacterium spp. and B. bacilliformis. The symptom frequencies were similar in patients diagnosed with Carrion’s disease and those co-infected with Mycobacterium spp. The most common symptoms were headaches, followed by malaise and arthralgia.

Objetivos: Describir las características clínico epidemiológicas de la Enfermedad de Carrión en pacientes que concurrieron al Instituto de Medicina Tropical Daniel Alcides Carrión durante el periodo 2010 al 2014. Material y métodos: Se analizó las historias clínicas de los pacientes que acudieron al IMT- UNMSM por sospecha de bartonelosis. Presenta un diseño no experimental tipo serie de casos, de modalidad retrospectiva, nivel de profundidad exploratoria, teniendo enfoque mixto (cuali-cuantitativo). Resultados: De 67 pacientes con sospecha de Bartonelosis el 90 % resultó con hemocultivo positivo. Del total de pacientes que acudieron al IMT – UNMSM el 42% fueron mujeres y 58% fueron hombres.La mayor cantidad de casos correspondieron a la fase aguda, encontrándose solo un caso en forma Verrucosa. Hubo 6 pacientes con frotis sanguíneo positivo, (3 mujeres y 3 hombres). El tiempo de enfermedad hasta que acuden al IMT-UNMSM fue: menor a 30 dias (28%), 1-6 meses (10%), mayor a 6 meses (18%), sin especificar (43%).El origende procedencia de contagio fue un 27% de Lima, 15% de Ancash, 8% de Junín, 7 % La Libertad, 5% Ucayali, 5% Piura y otros 33%. Los síntomas y signos más frecuentes que se encontraron en los pacientes con Bartonelosis fueron la fiebre en 48%, cefalea 34%, dolor articular 34%, palidez 21% y otros 37%. Conclusiones: Los pacientes que concurrieron al IMT- UNMSM para descartar, diagnosticar la Enfermedad de Carrión, al final representaron un gran porcentaje (90%) confirmados por hemocultivo y frotis sanguíneo. Existieron mayor porcentaje de hombres con Bartonelosis con respecto a mujeres. Los síntomas más frecuentes son fiebre 48%, cefalea 34%, dolor articular 34%.

Bartonella bacilliformis is a pathogen that is endemic in some areas of the Andean region of Peru, southern Ecuador and southern Colombia. This pathogen causes so-called Carrion's disease, a biphasic disease with acute and chronic phases (called Oroya fever and "Peruvian wart" respectively1-3). In the absence or delay of antibiotic treatment, the mortality rate in the acute phase is up to 88%1. The acute phase is characterised by fever and severe anaemia and may be followed, several weeks or months later, by the chronic eruptive phase due to endothelial cell proliferation2. No animal reservoir has been identified to date and it is considered that healthy carriers act as a pathogen reservoir in endemic areas