Background: In March 2013, the presence of an outbreak of Bartonella bacilliformis in the Rodriguez de Mendoza (Amazonas department, Peru) was reported. B. bacilliformis is an endemic pathogen of the Andean region, responsible for Carrion’s disease. One of the main problems of this illness is the lack of adequate technical and human resources for proper diagnosis in endemic rural areas. The objective of this study was to characterize a supposed B. bacilliformis outbreak, internationally informed in Rodriguez de Mendoza province. Methods & Materials: Fifty-three blood samples were recovered from people diagnosed with Carrion’s disease, either by optical microscopy and/or clinical manifestations. In all cases epidemiological and clinical data were recorded. The samples were cultured on Columbia Agar adding 10% of sheep blood and incubated at 28 ◦C for a period of 10 weeks. Every 14 days the plates were visually inspected to detect any bacterial growth. Additionally, the DNA was directly extracted from blood and 2 different 16S rRNA PCR schemes were used, one specific for Bartonella genus and other using universal primers. Twenty-six amplified products of universal 16S rRNA were randomly recovered and sequenced. Results: The main clinical presentations reported were headache (51%), physical discomfort (51%), chill (32%) and fever (24, 5%). Only 3 blood cultures were positive. No positive PCR was obtained when using the Bartonella specific PCR either on blood or on cultured bacteria. However, all the PCR with the universal primers were positive. The sequenced 26 (49%) samples were identified as Sphingomonas spp. being this microorganism the causative agent of this outbreak. In 17% of the cases, patients were reported to have aquatic activities. Conclusion: Several Sphingomonas spp. infections in humans have been reported, mostly limited to sporadic case reports or intra-hospitalary outbreaks, but as far as we know this is the first outbreak of Sphingomonas spp. described in a non-hospital environment. The association between 17% of patients with aquatic activities suggests that this was the most feasible transmission way. Training of health staff and development of new diagnostic able to be implemented in rural endemic areas is urgent in order to overcome wrong diagnostics and avoid wrong treatments.

Background: Bartonellagenus is a group of facultative intracellular pathogens that posses able to survive and proliferate inside of erythrocytes. Classified within this genus,Bartonella bacilliformisis of special relevance. This microorganism is the etiological agent of the so called Carrion’s Disease (Human bartonellosis). Additionally the presence of sub-clinical cases (asymptomatyc carriers) is of special interest, because acts as a reservoir of this illness. Carrión’s Disease is an endemic illnes in Perú, affecting in a special manner the north interandean valleys. However, the current in use diagnostic techniques (Giemsa Stain) possess low sensitivity and specificity, and due to the fact thatB. bacilliformispossess a low growth (weeks), bacterial cultures lacks of clinical utility. Thus suspictious cases frequently are not confirmed, and the real relevance of this illness remains underestimated. This work is addressed to the direct identification from blood samples ofBartonella baciliformisusing a conventional PCR. All patients were from the Cajamarca area being enrolled by the Epidemiological Surveillance program of DIRESA. Methods: The samples were processed at arriving to the laboratory, by molecular and microbiological techniques. Thus samples were cultured in Blood Columbia Agar (10%), in anaerobic conditions at 28 ◦C for a period of 2 months. Positive cultures were both Giemsa stained and identified by the amplification of a fragment the 16S rRNA gene. Genetic material was directly extracted from blood samples using the Kit High Pure (Roche diagnostic), and a fragment of 438 bp of the 16S rRNA gene was amplified withBartonellagenus specific primers. All positive PCR were sequenced (Macrogen-Korea). Results: A total of 134 blood samples were processed, from this 12 (8.9%) grown in blood agar, while in 18 (13.4%), including the aforementioned 12, the 16 s rRNA gene was amplified. In all cases the sequence analysis showed the presence ofB. bacilliformis Conclusion: Although microbiological culture is the gold standard in the identification ofBartonellaspp., this technique possess strong limitations due to the low growth of these microorganisms. However, the PCR is a rapid technique, possessing a high sensibility and specificity that may be used as routine diagnostic tool for the identification of Carrion’s Disease.

Background: In March 2013, the presence of an outbreak of Bartonella bacilliformis in the Rodriguez de Mendoza (Amazonas department, Peru) was reported. B. bacilliformis is an endemic pathogen of the Andean region, responsible for Carrion’s disease. One of the main problems of this illness is the lack of adequate technical and human resources for proper diagnosis in endemic rural areas. The objective of this study was to characterize a supposed B. bacilliformis outbreak, internationally informed in Rodriguez de Mendoza province. Methods & Materials: Fifty-three blood samples were recovered from people diagnosed with Carrion's disease, either by optical microscopy and/or clinical manifestations. In all cases epidemiological and clinical data were recorded. The samples were cultured on Columbia Agar adding 10% of sheep blood and incubated at 28°C for a period of 10 weeks. Every 14 days the plates were visually inspected to detect any bacterial growth. Additionally, the DNA was directly extracted from blood and 2 different 16S rRNA PCR schemes were used, one specific for Bartonella genus and other using universal primers. Twenty-six amplified products of universal 16S rRNA were randomly recovered and sequenced. Results: The main clinical presentations reported were headache (51%), physical discomfort (51%), chill (32%) and fever (24, 5%). Only 3 blood cultures were positive. No positive PCR was obtained when using the Bartonella specific PCR either on blood or on cultured bacteria. However, all the PCR with the universal primers were positive. The sequenced 26 (49%) samples were identified as Sphingomonas spp. being this microorganism the causative agent of this outbreak. In 17% of the cases, patients were reported to have aquatic activities. Conclusion: Several Sphingomonas spp. infections in humans have been reported, mostly limited to sporadic case reports or intra-hospitalary outbreaks, but as far as we know this is the first outbreak of Sphingomonas spp. described in a non-hospital environment. The association between 17% of patients with aquatic activities suggests that this was the most feasible transmission way.Training of health staff and development of new diagnostic able to be implemented in rural endemic areas is urgent in order to overcome wrong diagnostics and avoid wrong treatments.