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Se reporta la presencia de los flebotomos Lutzomyia hisuta hirsuta, Lu. nevesi, Lu. sherlocki, Lu. yuilli yuilli, Lu. migonei y Lu. sallesi en localidades contiguas a la ciudad de Tarapoto, Peru, ademas se registra la dominancia de Lu. nevesi como especie sinantropica de ambientes peri-domiciliarios.

Differences in the gut microbial content of Lutzomyia (Lu.) ayacuchensis, a primary vector of Andean-type cutaneous leishmaniasis in Ecuador and Peru, may influence the susceptibility of these sand flies to infection by Leishmania. As a first step toward addressing this hypothesis, a comparative analysis of bacterial and fungal compositions from Lu. ayacuchensis populations with differential susceptibilities to Leishmania was performed. Bacterial 16S rRNA gene amplification and Illumina MiSeq sequencing approaches were used to characterize the bacterial composition in wild-caught populations from the Andean areas of Ecuador and southern Peru at which the sand fly species transmit Leishmania (Leishmania) mexicana and Leishmania (Viannia) peruviana, respectively, and a population from the northern Peruvian Andes at which the transmission of Leishmania by Lu. ayacuchensis has not been reported. In the present study, 59 genera were identified, 21 of which were widely identified and comprised more than 95% of all bacteria. Of the 21 dominant bacterial genera identified in the sand flies collected, 10 genera had never been detected in field sand flies. The Ecuador and southern Peru populations each comprised individuals of particular genera, while overlap was clearly observed between microbes isolated from different sites, such as the number of soil organisms. Similarly, Corynebacterium and Micrococcus were slightly more dominant bacterial genera in the southern Peru population, while Ochrobactrum was the most frequently isolated from other populations. On the other hand, fungi were only found in the southern Peru population and dominated by the Papiliotrema genus. These results suggest that variation in the insect gut microbiota may be elucidated by the ecological diversity of sand flies in Peru and Ecuador, which may influence susceptibility to Leishmania infection. The present study provides key insights for understanding the role of the microbiota during the course of L. (L.) mexicana and L. (V.) peruviana infections in this important vector.

In the present study, we tested 391 fleas collected from guinea pigs (Cavia porcellus) (241 Pulex species, 110 Ctenocephalides felis, and 40 Tiamastus cavicola) and 194 fleas collected from human bedding and clothing (142 Pulex species, 43 C. felis, five T. cavicola, and four Ctenocephalides canis) for the presence of Bartonella DNA. We also tested 83 blood spots collected on Flinders Technology Associates (FTA) cards from guinea pigs inhabiting 338 Peruvian households. Bartonella DNA was detected in 81 (20.7%) of 391 guinea pig fleas, in five (2.6%) of 194 human fleas, and in 16 (19.3%) of 83 guinea pig blood spots. Among identified Bartonella species, B. rochalimae was the most prevalent in fleas (89.5%) and the only species found in the blood spots from guinea pigs. Other Bartonella species detected in fleas included B. henselae (3.5%), B. clarridgeiae (2.3%), and an undescribed Bartonella species (4.7%). Our results demonstrated a high prevalence of zoonotic B. rochalimae in households in rural areas where the research was conducted and suggested a potential role of guinea pigs as a reservoir of this bacterium.

The genus Bartonella encompasses several zoonotic species that cause emerging infectious diseases in humans. These species are transmitted to humans by hematophagous arthropods. In South America, sand flies are the most important vectors of Bartonella bacilliformis, the etiological agent of Verruga peruana. Due to the importance of bartonellosis in public health, the aim of this study was to detect Bartonella in wild populations of phlebotomine sand flies in southern Mexico. In total, 29 sand flies belonging to 6 species were collected. Only 2 specimens were positive, representing a prevalence of 8.69% (2/23). This appears to be the first report of Bartonella sp. in phlebotomine sand flies outside the endemic area of Verruga peruana in South America. Further studies are needed to clarify the relation between this bacterium and sand flies in Mexico.

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mannose phosphate isomerase (mpi) gene was applied to 134 skin samples collected from patients with cutaneous leishmaniasis (CL) in Peru for identification of the infecting parasite at the species level, and the results were compared with those of cytochrome b (cyt b) gene sequencing obtained in previous studies. Although most results (121/134) including 4 hybrids of Leishmania (Viannia) braziliensis and L. (V.) peruviana corresponded to those obtained in the previous study, PCR-RFLP analyses revealed the distribution of putative hybrid strains between L. (V.) peruviana and L. (V.) lainsoni in two samples, which has never been reported. Moreover, parasite strains showing discordance between kinetoplast and nuclear genes (kDNA and nDNA), so-called mito-nuclear discordance, were identified in 11 samples. Of these, six strains had the kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) guyanensis, and three strains had the kDNAs of L. (V.) shawi and nDNAs of L. (V.) braziliensis. The rest were identified as mito-nuclear discordance strains having kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) lainsoni, and kDNAs of L. (V.) lainsoni and nDNAs of L. (V.) braziliensis. The results demonstrate that Leishmania strains in Peru are genetically more complex than previously considered., Author summary Protozoan parasites of the genus Leishmania are able to undergo genetic exchange during their growth. The previous description of hybrids in Peru and the recent discovery of unexpected genetically complex strains having characteristics of both hybrid and mito-nuclear discordance in its neighbouring country (Ecuador) with a similar eco-epidemiological situation led us to consider that the genetic structure of Leishmania strains in Peru is more complicated than previously thought. In an effort to revise the data on Leishmania strain dispersion in Peru and to search for evidence of genetic recombination, the present study was conducted. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis targeting the mannose phosphate isomerase (mpi) gene sequence was performed to identify the infecting parasite at the species level in 134 skin samples collected from patients with cutaneous leishmaniasis (CL) in Peru, and the results were compared with those of cytochrome b (cyt b) gene sequencing obtained in previous studies. Most results (121/134) including 4 hybrids between L. (V.) braziliensis and L. (V.) peruviana showed agreement between PCR-RFLP of the mpi gene and cyt b gene sequence analysis; however, 13 of 134 samples revealed the distribution of strains with hybrids and mito-nuclear discordance. The results demonstrate that genetically complex Leishmania strains are present in Peru. These findings indicate that Leishmania strain dispersion in Peru is genetically more complex than previously considered. Further prospective studies including larger samples and the isolation of parasite strains are required to update the available data.

This study provides comprehensive information on the past and current status of the Andean cutaneous leishmaniasis (Andean-CL, uta) in Peru and Ecuador, mainly focusing on the causative Leishmania parasites and clinico-epidemiological features. Available information and data including our unpublished works were analyzed thoroughly. Endemic regions of the Andean-CL (uta) in Peru run from the north Piura/Cajamarca to the south Ayacucho at a wide range of the Pacific watersheds of the Andes through several departments, while in Ecuador those exist at limited and spotted areas in the country's mid-southwestern two provinces, Azuay and Chimborazo. The principal species of the genus Leishmania are completely different at subgenus level, L. (Viannia) peruviana in Peru, and L. (Leishmania) mexicana and L. (L.) major-like (infrequent occurrence) in Ecuador. The Peruvian uta is now prevalent in different age and sex groups, being not clearly defined as found in the past. The precise reasons are not known and should be elucidated further, though probable factors, such as emergence of other Leishmania parasites, non-immune peoples' migration into the areas, etc., were discussed briefly in the text. The Andean-CL cases in Ecuador are more rural than before, probably because of a rapid development of the Leishmania-positive communities and towns, and the change of life-styles of the inhabitants, including newly constructed houses and roads in the endemic areas. Such information is helpful for future management of the disease, not only for Leishmania-endemic areas in the Andes but also for other endemic areas.

The Peruvian Andes presents a climate suitable for many species of sandfly that are known vectors of leishmaniasis or bartonellosis, including Lutzomyia peruensis (Diptera: Psychodidae), among others. In the present study, occurrences data for Lu. peruensis were compiled from several items in the scientific literature from Peru published between 1927 and 2015. Based on these data, ecological niche models were constructed to predict spatial distributions using three algorithms [Support vector machine (SVM), the Genetic Algorithm for Rule-set Prediction (GARP) and Maximum Entropy (MaxEnt)]. In addition, the environmental requirements of Lu. peruensis and three niche characteristics were modelled in the context of future climate change scenarios: (a) potential changes in niche breadth; (b) shifts in the direction and magnitude of niche centroids, and (c) shifts in elevation range. The model identified areas that included environments suitable for Lu. peruensis in most regions of Peru (45.77%) and an average altitude of 3289 m a.s.l. Under climate change scenarios, a decrease in the distribution areas of Lu. peruensis was observed for all representative concentration pathways. However, the centroid of the species' ecological niche showed a northwest direction in all climate change scenarios. The information generated in this study may help health authorities responsible for the supervision of strategies to control leishmaniasis to coordinate, plan and implement appropriate strategies for each area of risk, taking into account the geographic distribution and potential dispersal of Lu. peruensis.

In the present study, we tested 391 fleas collected from guinea pigs (Cavia porcellus) (241 Pulex species, 110 Ctenocephalides felis, and 40 Tiamastus cavicola) and 194 fleas collected from human bedding and clothing (142 Pulex species, 43 C. felis, five T. cavicola, and four Ctenocephalides canis) for the presence of Bartonella DNA. We also tested 83 blood spots collected on Flinders Technology Associates (FTA) cards from guinea pigs inhabiting 338 Peruvian households. Bartonella DNA was detected in 81 (20.7%) of 391 guinea pig fleas, in five (2.6%) of 194 human fleas, and in 16 (19.3%) of 83 guinea pig blood spots. Among identified Bartonella species, B. rochalimae was the most prevalent in fleas (89.5%) and the only species found in the blood spots from guinea pigs. Other Bartonella species detected in fleas included B. henselae (3.5%), B. clarridgeiae (2.3%), and an undescribed Bartonella species (4.7%). Our results demonstrated a high prevalence of zoonotic B. rochalimae in households in rural areas where the research was conducted and suggested a potential role of guinea pigs as a reservoir of this bacterium.

The genus

To obtain further insight into geographic distribution of Leishmania species in Peru, a countrywide survey, including central to southern rainforest areas where information on causative parasite species is limited, was performed based on cytochrome b (cyt b) and mannose phosphate isomerase (mpi) gene analyses. A total of 262 clinical samples were collected from patients suspected of cutaneous leishmaniasis (CL) in 28 provinces of 13 departments, of which 99 samples were impregnated on FTA (Flinders Technology Associates) cards and 163 samples were Giemsa-stained smears. Leishmania species were successfully identified in 83 (83.8%) of FTA-spotted samples and 59 (36.2%) of Giemsa-stained smear samples. Among the 142 samples identified, the most dominant species was Leishmania (Viannia) braziliensis (47.2%), followed by L. (V.) peruviana (26.1%), and others were L. (V.) guyanensis, L. (V.) lainsoni, L. (V.) shawi, a hybrid of L. (V.) braziliensis and L. (V.) peruviana, and Leishmania (Leishmania) amazonensis. Besides the present epidemiological observations, the current study provided the following findings: 1) A hybrid of L. (V.) braziliensis and L. (V.) peruviana is present outside the Department of Huanuco, the only place reported, 2) Many cases of CL due to L. (V.) lainsoni, an uncommon causative species in Peru, were observed, and 3) L. (V.) shawi is widely circulating in southern Amazonian areas in Peru., Author summary Leishmaniasis, a neglected tropical disease (NTD) caused by the intracellular protozoa of the genus Leishmania, affects at least 12 million people in 96 countries. Peru is one of the most highly endemic countries for cutaneous leishmaniasis (CL), and our previous study identified Leishmania (Viannia) braziliensis, L. (V.) peruviana, and L. (V.) guyanensis in the tropical rainforest, in the Andean highlands, and in the northern and central rainforest areas, respectively, as the main CL-causative agents. In addition, distribution of L. (V.) lainsoni, L. (V.) shawi, a hybrid of L. (V.) braziliensis and L. (V.) peruviana, and Leishmania (Leishmania) amazonensis has been identified. Of these, one case each of L. (V.) shawi infection was reported from the Departments of Junin and Madre de Dios, while clinical cases due to the hybrid of L. (V.) braziliensis and L. (V.) peruviana were recorded only in the Department of Huanuco. To further elucidate the current geographic distribution of causative Leishmania species in Peru, a countrywide survey, including central to southern rainforest areas where little information on causative parasites is available, was performed based on the cytochrome b (cyt b) gene sequence and PCR-RFLP analysis of the mannose phosphate isomerase (mpi) gene by using FTA (Flinders Technology Associates) card-spotted samples and smear slides as DNA sources. In addition to current epidemiological observations, the current study revealed that 1) A hybrid of L. (V.) braziliensis and L. (V.) peruviana was identified, for the first time, outside the Department of Huanuco, 2) L. (V.) lainsoni, an uncommon CL-causative species in Peru, was predominantly found in the Department of Puno, where causative Leishmania species are not well-studied, and 3) New endemic foci of L. (V.) shawi were identified in central to southern rainforest areas of Peru.

The natural infection of sand flies by Leishmania was investigated in Andean areas located between the Central and Eastern Cordilleras of northern Peru where cutaneous leishmaniasis caused by Leishmania (Viannia) peruviana is endemic. Sand flies were captured at five locations along the Utcubamba River in the Department of Amazonas, and morphologically identified under a microscope. Among 422 female sand flies dissected, the most dominant species was Pintomyia verrucarum (320 flies), followed by Pi. maranonensis (83 flies), Pi. robusta (13 flies), and Lutzomyia castanea (6 flies). Genetic analysis of sand flies from these areas together with those from other areas revealed that individuals of Pi. verrucarum were closely related regardless of morphological variation of their spermathecae. On the other hand, individuals of Pi. maranonensis collected in the study area were distant from those of other areas with genetic distances over the intraspecific level but mostly below the interspecific level, suggesting the unique characteristics of sand flies in this area. The natural infection of sand flies by flagellate parasites was detected mainly in the hindgut of each one of Pi. verrucarum and Pi. maranonensis. Both parasite species were identified as L. (V.) peruviana based on cytochrome b and mannose phosphate isomerase gene analyses. In addition, parasite species obtained from the lesion of a patient with cutaneous leishmaniasis in the study area in this period was identified as L. (V.) peruviana. These results strongly suggest that Pi. verrucarum and Pi. maranonensis are responsible for the transmission of L. (V.) peruviana in these areas. This is the first report of the natural infection of Pi. maranonensis by L. (V.) peruviana., Author summary Phlebotomine sand flies are tiny insects of the family Psychodidae in the order Diptera, and female sand flies suck blood for egg production. Approximately 1,020 sand fly species have been recorded in the world, of which about 550 species are in the New World. Only a part of them are associated with medically important infectious diseases such as leishmaniasis, and importantly, each vector species transmits specific species of Leishmania. Since the infecting Leishmania species is the major determinant of the clinical outcome and its endemicity is largely dependent on the prevalence of the vector species, the identification of circulating sand flies and vector species, which determine transmissible parasite species, is important to predict the risk and expansion of the disease in endemic and surrounding areas. However, the vector species involved in disease transmission remains unidentified in most endemic areas because the infection rate in sand fly populations is very low. In the present study, sand flies were investigated in the Department of Amazonas in the Eastern Andes of northern Peru, in which cutaneous leishmaniasis caused by Leishmania (Viannia) peruviana is endemic, to clarify the transmission mechanism of leishmaniasis in these areas. In addition, genetic analyses of circulating sand flies were performed to elucidate the characteristics of sand flies in these areas.

Leishmaniasis remains one of the world's most neglected diseases, and early detection of the infectious agent, especially in developing countries, will require a simple and rapid test. In this study, we established a quick, one-step, single-tube, highly sensitive loop-mediated isothermal amplification (LAMP) assay for rapid detection of Leishmania DNA from tissue materials spotted on an FTA card. An FTA-LAMP with pre-added malachite green was performed at 64 degrees C for 60 mm using a heating block and/or water bath and DNA amplification was detected immediately after incubation. The LAMP assay had high detection sensitivity down to a level of 0.01 parasites per mu l. The field- and clinic-applicability of the colorimetric FTA-LAMP assay was demonstrated with 122 clinical samples collected from patients suspected of having cutaneous leishmaniasis in Peru, from which 71 positives were detected. The LAMP assay in combination with an FTA card described here is rapid and sensitive, as well as simple to perform, and has great potential usefulness for diagnosis and surveillance of leishmaniasis in endemic areas. (C) 2015 Elsevier B.V. All rights reserved.

Identification of mosquito species is necessary for determining the entomological components of malaria transmission, but it can be difficult in morphologically similar species. DNA sequences are largely used as an additional tool for species recognition, including those that belong to species complexes. Kerteszia mosquitoes are vectors of human and simian malaria in the Neotropical Region, but there are few DNA sequences of Kerteszia species in public databases. In order to provide relevant information about diversity and improve knowledge in taxonomy of Kerteszia species in Peru, we sequenced part of the mitochondrial genome, including the cytochrome c oxidase I (COI) barcode region. Phylogenetic analyses structured all species of mosquitoes collected in Peru into a single clade, separate from the Brazilian species. The Peruvian clade was composed of two lineages, encompassing sequences from Anopheles Kerteszia boliviensis and Anopheles Kerteszia pholidotus. An. pholidotus sequences were recorded for the first time in Peru, whereas An. boliviensis sequences were for the first time published in the GenBank database. Sequences generated from specimens morphologically identified as Anopheles Kerteszia cruzii clustered into three separate clades according to the collection localities of Serra do Mar, Serra da Mantiqueira, and Serra da Cantareira, confirming An. cruzii as a species complex, composed of at least three putative species.

Background; The goal of this study was to reassess the taxonomic status of A. maculatum, A. triste and A. tigrinum by phylogenetic analysis of five molecular markers [four mitochondrial: 12S rDNA, 16S rDNA, the control region (DL) and cytochrome c oxidase 1 (cox1), and one nuclear: ribosomal intergenic transcribed spacer 2 (ITS2)]. In addition, the phenotypic diversity of adult ticks identified as A. maculatum and A. triste from geographically distinct populations was thoroughly re-examined. Results: Microscopic examination identified four putative morphotypes distinguishable by disjunct geographical ranges, but very scant fixed characters. Analysis of the separated mitochondrial datasets mostly resulted in conflicting tree topologies. Nuclear gene sequences were almost identical throughout the geographical ranges of the two species, suggesting a very recent, almost explosive radiation of the terminal operational taxonomic units. Analysis of concatenated molecular datasets was more informative and indicated that, although genetically very close to the A. maculatum - A. triste lineage, A. tigrinum was a monophyletic separate entity. Within the A. maculatum - A. triste cluster, three main clades were supported. The two morphotypes, corresponding to the western North American and eastern North American populations, consistently grouped in a single monophyletic clade with many shared mitochondrial sequences among ticks of the two areas. Ticks from the two remaining morphotypes, south-eastern South America and Peruvian, corresponded to two distinct clades. Conclusions: Given the paucity of morphological characters, the minimal genetic distance separating morphotypes, and more importantly the fact that two morphotypes are genetically indistinguishable, our data suggest that A. maculatum and A. triste should be synonymized and that morphological differences merely reflect very recent local adaptation to distinct environments in taxa that might be undergoing the first steps of speciation but have yet to complete lineage sorting. Nonetheless, future investigations using more sensitive nuclear markers and/or crossbreeding experiments might reveal the occurrence of very rapid speciation events in this group of taxa. Tentative node dating revealed that the A. tigrinum and A. maculatum - A. triste clades split about 2 Mya, while the A. maculatum - A.triste cluster radiated no earlier than 700,000 years ago. EEA Rafaela Fil: Lado, Paula. Georgia Southern University. Institute for Coastal Plain Science. United States National Tick Collection; Estados Unidos Fil: Nava, Santiago. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Mendoza-Uribe, Leonardo. Instituto Nacional de Salud. Laboratorio de Entomología; Perú Fil: Cáceres, Abraham G. Universidad Nacional Mayor de San Marcos. Facultad de Medicina. Departamento Académico de Microbiología Médica; Perú. Instituto Nacional de Salud. Laboratorio de Entomología; Peru Fil: Delgado de la Mora, Jesus. University of Sonora. Department of Medicine and Health Sciences; México Fil: Licona Enriquez, Jesus D. University of Sonora. Department of Medicine and Health Sciences; México Fil: Delgado de la Mora, David. Technologic Institute of Sonora. Department of Agronomic and Veterinary Sciences; México Fil: Labruna, Marcelo B. Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia. Departamento de Medicina Veterinária Preventiva e Saúde Animal; Brasil Fil: Durden, Lance A. Georgia Southern University. Department of Biology; Estados Unidos Fil: Allerdice, Michelle E. J. Centers for Disease Control and Prevention. National Center for Emerging and Zoonotic Infectious Diseases. Rickettsial Zoonoses Branch; Estados Unidos Fil: Paddock, Christopher D. Centers for Disease Control and Prevention. National Center for Emerging and Zoonotic Infectious Diseases. Rickettsial Zoonoses Branch; Estados Unidos Fil: Szabó, Matías P.J. Universidade Federal de Uberlandia. Faculdade de Medicina Veterinaria e Zootecnia. Laboratório de Ixodologia; Brasil Fil: Venzal, José Manuel. Universidad de la República. CENUR Litoral Norte-Salto. Facultad de Veterinaria. Departamento de Parasitología Veterinaria; Uruguay Fil: Guglielmone, Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Beati, Lorenza. Georgia Southern University. Institute for Coastal Plain Science. United States National Tick Collection; Estados Unidos

In the present study, we investigated 238 fleas collected from cats and dogs in three regions of Peru (Ancash, Cajamarca, and Lima) for the presence of Bartonella DNA. Bartonella spp. were detected by amplification of the citrate synthase gene (16.4%) and the 16S-23S intergenic spacer region (20.6%). Bartonella rochalimae was the most common species detected followed by Bartonella clarridgeiae and Bartonella henselae. Our results demonstrate that dogs and cats in Peru are infested with fleas harboring zoonotic Bartonella spp. and these infected fleas could pose a disease risk for humans.

An analysis of reported cases of cutaneous leishmaniasis (CL) was performed using the data registered in the southern Ecuadorian Amazon region during 27 years from 1986 to 2012. The cases/subjects with both the suspected CL lesions and the amastigote-positive results were recruited for the analysis. The yearly occurrence of cases showed a markedly higher number during the six years, 1988 and 1993. After 1994 when the insecticide spraying campaign using helicopter in 1993-1994, the number dropped remarkably. Then, the yearly occurrence gradually fluctuated from 101 cases in 1996 to 11 in 2009, maintaining a low number of cases after the campaign. The monthly occurrence of cases showed a markedly high number during March and August, suggesting a correlation to the rainy season (months) in the areas. A statistical significance was found between the monthly average number of the CL case and the average precipitation (p=0.01474). It was suggested that the time of transmission of CL would depend on the rainy seasons at each endemic area of Ecuador, which has a diverse climatic feature depending on the geographic regions. Such information at given leishmaniasis-endemic areas of Ecuador would be important for the future planning of the disease control. Molecular analysis and characterization of clinical samples revealed the presence of Leishmania (Viannia) braziliensis.

Phlebotomine sand flies are the only proven vectors of leishmaniases, a group of human and animal diseases. Accurate knowledge of sand fly species identification is essential in understanding the epidemiology of leishmaniasis and vector control in endemic areas. Classical identification of sand fly species based on morphological characteristics often remains difficult and requires taxonomic expertise. Here, we generated DNA barcodes of the cytochrome c oxidase subunit 1 (COI) gene using 159 adult specimens morphologically identified to be 19 species of sand flies, belonging to 6 subgenera/species groups circulating in Peru, including the vector species. Neighbor-joining (NJ) analysis based on Kimura 2-Parameter genetic distances formed non-overlapping clusters for all species. The levels of intraspecific genetic divergence ranged from 0 to 5.96%, whereas interspecific genetic divergence among different species ranged from 8.39 to 19.08%. The generated COI barcodes could discriminate between all the sand fly taxa. Besides its success in separating known species, we found that DNA barcoding is useful in revealing population differentiation and cryptic diversity, and thus promises to be a valuable tool for epidemiological studies of leishmaniasis. Copyright (C) 2015 Published by Elsevier B.V. All rights reserved.

Haplotype and gene network analyses were performed on mitochondrial cytochrome oxidase I and cytochrome b gene sequences of Lutzomyia (Lu.) ayacuchensis populations from Andean areas of Ecuador and southern Peru where the sand fly species transmit Leishmania (Leishmania) mexicana and Leishmania (Viannia) peruviana, respectively, and populations from the northern Peruvian Andes, for which transmission of Leishmania by Lu. ayacuchensis has not been reported. The haplotype analyses showed higher intrapopulation genetic divergence in northern Peruvian Andes populations and less divergence in the southern Peru and Ecuador populations, suggesting that a population bottleneck occurred in the latter populations, but not in former ones. Importantly, both haplotype and phylogenetic analyses showed that populations from Ecuador consisted of clearly distinct clusters from southern Peru, and the two populations were separated from those of northern Peru. Crown Copyright (C) 2014 Published by Elsevier B.V. All rights reserved.

The vector Lutzomyia sand flies and reservoir host mammals of the Leishmania parasites, causing the Andean cutaneous leishmaniasis (Andean-CL, uta) in Peru and Ecuador were thoroughly reviewed, performing a survey of literatures including our unpublished data. The Peruvian L. (V.) peruviana, a principal Leishmania species causing Andean-CL in Peru, possessed three Lutzomyia species, Lu. peruensis, Lu. verrucarum and Lu. ayacuchensis as vectors, while the Ecuadorian L. (L.) mexicana parasite possessed only one species Lu. ayacuchensis as the vector. Among these, the Ecuadorian showed a markedly higher rate of natural Leishmania infections. However, the monthly and diurnal biting activities were mostly similar among these vector species was in both countries, and the higher rates of infection (transmission) reported, corresponded to sand fly's higher monthly-activity season (rainy season). The Lu. tejadai sand fly participated as a vector of a hybrid parasite of L. (V.) braziliensis/L. (V.) peruviana in the Peruvian Andes. Dogs were considered to be principal reservoir hosts of the L. (V.) peruviana and L. (L.) mexicana parasites in both countries, followed by other sylvatic mammals such as Phyllotis andium, Didelphis albiventris and Akodon sp. in Peru, and Rattus rattus in Ecuador, but information on the reservoir hosts/mammals was extremely poor in both countries. Thus, the Peruvian disease form demonstrated more complicated transmission dynamics than the Ecuadorian. A brief review was also given to the control of vector and reservoirs in the Andes areas. Such information is crucial for future development of the control strategies of the disease.

Entomological monitoring of Leishmania infection in leishmaniasis endemic areas offers epidemiologic advantages for predicting the risk and expansion of the disease, as well as evaluation of the effectiveness of control programs. In this study, we developed a highly sensitive loop-mediated isothermal amplification (LAMP) method for the mass screening of sand flies for Leishmania infection based on the 18S rRNA gene. The LAMP technique could detect 0.01 parasites, which was more sensitive than classical PCR. The method was robust and could amplify the target DNA within 1 h from a crude sand fly template without DNA purification. Amplicon detection could be accomplished by the newly developed colorimetric malachite green (MG)—mediated naked eye visualization. Pre-addition of MG to the LAMP reaction solution did not inhibit amplification efficiency. The field applicability of the colorimetric MG-based LAMP assay was demonstrated with 397 field-caught samples from the endemic areas of Ecuador and eight positive sand flies were detected. The robustness, superior sensitivity, and ability to produce better visual discriminatory reaction products than existing LAMP fluorescence and turbidity assays indicated the field potential usefulness of this new method for surveillance and epidemiological studies of leishmaniasis in developing countries.

Introducción. Trypanosoma cruzi se ha dividido en seis unidades taxonómicas discretas (Discreet Typing Units, DTU) denominadas TcI, TcII, TcIII, TcIV, TcV y TcVI. Aún se desconocen los factores determinantes de la dinámica de la transmisión vectorial de los genotipos de T. cruzi en las diferentes regiones geográficas de distribución de la enfermedad de Chagas en Perú.Objetivo. Detectar y tipificar las unidades taxonómicas discretas de T. cruzi en las heces de siete especies de triatominos (Panstrongylus chinai, P. geniculatus, P. herreri, Rhodnius robustus, R. pictipes, Triatoma carrioni y T. infestans), capturados en ocho departamentos de diferentes regiones naturales de Perú.Materiales y métodos. Se examinaron 197 insectos para la detección de tripanosomas. Se extrajo el ADN del contenido intestinal de cada insecto y se amplificó mediante reacción en cadena de la polimerasa (PCR) de los genes kDNA, SL-IR, 24Sα rRNA y 18Sα RNA para detectar las DTU de T. cruzi. Resultados. Se detectaron cinco infecciones con T. rangeli y 113 con T. cruzi. De estas últimas, fue posible identificar 95 de TcI (dos en P. chinai, una en P. geniculatus, 68 en P. herreri, cuatro en R. pictipes, siete en R. robustus, una en T. carrioni, y 12 en T. infestans); cinco de TcII (cuatro en P. herreri, una en T. infestans); cuatro de TcIII (tres en P. herreri, una en R. robustus) y cuatro infecciones de TcIV en P. herreri.Conclusión. Este es el primer trabajo de caracterización a gran escala de T. cruzi en el intestino de vectores de importancia epidemiológica en Perú, orientado a generar información básica que permita entender la dinámica de la transmisión vectorial de T. cruzi en esta región del continente.

A PCR-Restriction Fragment Length Polymorphism (RFLP) targeting the mannose phosphate isomerase gene was established to differentiate Leishmania species distributed near the Department of Huanuco, Peru. The technique was applied to 267 DNA samples extracted from Giemsa-stained smears of cutaneous lesions taken from patients suspected for cutaneous leishmaniasis in the area, and the present status of causative Leishmania species was identified. Of 114 PCR-amplified samples, 22, 19, 24 and 49 samples were identified to be infected by Leishmania (Viannia) braziliensis, L. (V.) peruviana, L. (V.) guyanensis, and a hybrid of L. (V.) braziliensis/L. (V.) peruviana, respectively, and the validity of PCR-RFLP was confirmed by sequence analysis. Since PCR-RFLP is simple and rapid, the technique will be a useful tool for the epidemiological study of leishmaniasis.

A total of 103 ticks, collected from canines, horses, donkeys, and snakes from Peru, were screened for the presence of Bartonella DNA by polymerase chain reaction analysis. Bartonella DNA was detected in two ticks using Bartonella 16S-23S intergenic spacer region primers and in an additional two ticks using Bartonella NADH dehydrogenase gamma subunit gene (nuoG) primers. Bartonella rochalimae Eremeeva et al., B. quintana Schmincke, and B. elizabethae Daly et al. DNA was detected in a Rhipicephalus sanguineus Latreille (Acari: Ixodidae) female tick removed from a dog and B. quintana DNA was present in a Dermacentor nitens Neumann (Acari: Ixodidae) pool of five larvae, one nymph, and one adult male tick collected from donkeys. This is the first study to report the detection of B. rochalimae, B. quintana, and B. elizabethae DNA in ticks from Peru. Further investigations must be performed to decipher the role ticks may play in the transmission of Bartonella species.

Within the sand fly genus Lutzomyia, the Verrucarum species group contains several of the principal vectors of American cutaneous leishmaniasis and human bartonellosis in the Andean region of South America. The group encompasses 40 species for which the taxonomic status, phylogenetic relationships, and role of each species in disease transmission remain unresolved. Mitochondrial cytochrome c oxidase I (COI) phylogenetic analysis of a 667-bp fragment supported the morphological classification of the Verrucarum group into series. Genetic sequences from seven species were grouped in well-supported monophyletic lineages. Four species, however, clustered in two paraphyletic lineages that indicate conspecificity--the Lutzomyia longiflocosa-Lutzomyia sauroida pair and the Lutzomyia quasitownsendi-Lutzomyia torvida pair. COI sequences were also evaluated as a taxonomic tool based on interspecific genetic variability within the Verrucarum group and the intraspecific variability of one of its members, Lutzomyia verrucarum, across its known distribution.

The natural infection of sand flies by Leishmania species was studied in the Andean areas of Peru where cutaneous leishmaniasis caused by Leishmania (Viannia) peruviana is endemic. Sand flies were captured by human bait and Center for Disease Control (CDC) light trap catches at Nambuque and Padregual, Department of La Libertad, Peru, and morphologically identified. Among 377 female sand flies dissected, the two dominant man-biting species were Lutzomyia (Helcocyrtomyia) peruensis (211 flies) and Lutzomyia (Helcocyrtomyia) caballeroi (151 flies). Another sand fly species captured by light trap was Warileya phlebotomanica (15 flies). The natural infection of sand flies by flagellates was detected in 1.4% of Lu. (H.) peruensis and 2.6% of Lu. (H.) caballeroi, and the parasite species were identified as Le. (V.) peruviana and Trypanosoma avium, respectively, by molecular biological methods. The results indicated that the vector species responsible for the transmission of leishmaniasis in the study areas is Lu. (H.) peruensis. In addition, the presence of Trypanosoma in man-biting sand fly species means that more careful consideration is necessary for vector research in areas of Andean Peru where leishmaniasis is endemic.

In this study, each of 60 rRNA internal transcribed spacer (ITS) 1 and ITS2 sequences was determined from 44 individuals of 14 morphologically identified New World sand fly Lutzomyia species in Ecuador, and their interspecies and intraspecies genetic diversity was compared. Distinguishing between related species based on the ITS1 sequence was difficult because of variability, while the genetic diversity of ITS2 was distinct even among closely related species. Further, an assessment of intraspecies ITS sequence diversity in the subgenus Helcocyrtomyia revealed no correlation between sequence variation and geographic distribution. The results strongly suggested ITS2 to be a more suitable marker than ITS1 for the taxonomic analysis of Lutzomyia species including closely related species. Moreover, neither ITS sequence may be useful for the analysis of population structures in Lutzomyia species.

A male of a new species Micropygomyia (Micropygomyia) ancashensis sp. nov. and a female of the Lutzomyia (Helcocyrtomyia) chavinensis Pérez & Ogusuku, 1999 captured in the Antonio Raymondi province, department of Ancash, Peru are described and illustrated. The new species belongs to the cayennensis series, being closest to Mi. lewisi (Feliciangeli Ordoñez & Férnandez) though presenting highly sclerotized pseudotracheae on the labella that seem to constitute an autapomorphy among American Phlebotominae, and a clypeus almost completely covered with setae, as in Warileya. The female of Lu. chavinensis is close to those of Lu. blancasi Galati & Cáceres 1990, Lu. noguchii (Shannon, 1929), and Lu. pallidithorax Galati & Cáceres 1994.

The natural infection of sand flies by Leishmania was examined in the Department of Huanuco of Peru, where cutaneous leishmaniasis caused by a hybrid of Leishmania (Viannia) braziliensis/L. (V.) peruviana is endemic. A total of 2,997 female sand flies were captured by CDC light traps and Shannon traps, of which 2,931 and 66 flies were identified as Lutzomyia tejadai and Lu fischeri, respectively. Using crude DNA extracted from individual sand flies as a template, Leishmania DNA was detected from one Lu. tejadai. The parasite species was identified as a hybrid of L. (V.) braziliensis/L. (V.) peruviana on the basis of cytochrome b and mannose phosphate isomerase gene analyses. The result suggested that Lu. tejadai is responsible for the transmission of the hybrid Leishmania circulating in this area., Author Summary Leishmaniasis is a protozoan disease caused by members of the genus Leishmania, which are distributed worldwide, especially in tropical and subtropical areas. More than 20 species of Leishmania are described as causative agents of human leishmaniases and clinical features are largely associated with the infective species. The parasites are transmitted by bites of female sand flies of the genus Phlebotomus in the Old World and Lutzomyia in the New World. Approximately 800 sand fly species have been recorded; of these, fewer than 10% have been confirmed as vectors of Leishmania and only a restricted number of species transmit specific Leishmania species. Therefore, sand fly surveillance is important for predicting risk and spread of leishmaniasis in endemic and surrounding areas. Cutaneous leishmaniasis caused by a hybrid of L. (Viannia) braziliensis/L. (V.) peruviana have been reported in the Department of Huanuco of Peru; however, the vector species has not been identified. In the present study, approximately 3,000 sand flies were examined, and the sand fly infected by the hybrid Leishmania strain was successfully identified.

A new species of phlebotomine, Lutzomyia (Helcocyrtomyia) herreri sp. nov., belonging to the series osornoi, from the Department of Puno, Peru, is described. Identification keys for both sexes of the species of this series are presented.

Lutzomyia chotensis sp. nov. (Diptera, Psychodidae, Phlebotominae) from Cajamarca Department, Peru, is described and its taxonomic position discussed.

A reassessment of the taxonomic status of Amblyomma cajennense based on the morphological analyses of ticks from the whole distribution area of the species resulted in the redescription of A. cajennense, the validation of 2 species which had been reduced to synonymy in the past, Amblyomma mixtum and Amblyomma sculptum, and the description and definition of 3 new species, Amblyomma tonelliae n. sp., Amblyomma interandinum n. sp., and Amblyomma patinoi n. sp. This study provides descriptions and redescriptions, scanning electron microscopic and stereomicroscopic images, updated synonymies, information on geographical distributions, and host associations for each of the 6 species. Amblyomma cajennense s.s. is found in the Amazonian region of South America, A. interandinum is reported from the northern part of the Inter-Andean valley of Peru, A. mixtum is present from Texas (U.S.A.) to western Ecuador, A. patinoi occurs in the Eastern Cordillera of Colombia, A. tonelliae is associated with the dry areas of the Chaco region which spans from central-northern Argentina to Bolivia and Paraguay, whereas A. sculptum is distributed from the humid areas of northern Argentina, to the contiguous regions of Bolivia and Paraguay and the coastal and central-western states of Brazil. Fil: Nava, Santiago. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Lorenza, Beati. Georgia Southern University. Institute for Coastal Plain Science; Estados Unidos Fil: Labruna, Marcelo B.. Universidade de Sao Paulo; Brasil Fil: Cáceres, Abraham G.. Universidad Nacional Mayor de San Marcos; Perú Fil: Mangold, Atilio Jose. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina Fil: Guglielmone, Alberto Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina

A las 120 especies de flebotomíneos reportadas hasta 1999 para el Perú, se adiciona: Brumptomyia quimperi, Evandromyia sallesi, Lutzomyia scorsai, L. rispaili, L. wattsi, Micropygomyia longipennis, Trichophoromyia howardi, T. arevaloi, Trichopygomyia turelli, W. euniceae y W leponti, incrementando a 131 especies el número de flebotomíneos. Se da a conocer la distribución geográfica de flebotomíneos en los departamentos de Huánuco, Pasco y Cusco; además, se destaca nuevos reportes de flebotomíneos en los departamentos de Ayacucho, Amazonas, Cajamarca, Huancavelica, Junín, La Libertad, Loreto y San Martín. Así mismo, se presentan evidencias epidemiológicas de algunos flebotomineos de los subgéneros Helcocyrtomyia y Pifanomyia como probables vectores de agentes patógenos de la leishmaniosis tegumentaria y de la enfermedad de Carrión (verruga peruana)., One hundred and twenty species had been reported up to 1999 and the following 11 are now added to Peruvian phlebotomine fauna: Brumptomyia quimperi, Evandromyia sallesi, Lutzomyia fispaili, L. scorzai, L. watsi, Micropygomyia longipennis, Trichophoromyia howardi, T. arevaloi, Trichopygomyia turelli, Warileya euniceae and W. leponti. A complete list of species in Huanuco, Pasco and Cusco departments is included and new records for Ayacucho, Amazonas, Cajamarca, Huancavelica, Junin, La Libertad, Loreto and San Martin are given. There appears to be epidemiological evidences of some phlebotomines of the subgenera Helcocyrtomyia and Pifanomyia as likely vectors of dermal leishmaniosis and Carrion's disease.

A new species of Culicoides of the subgenus Mataemyia Vargas, Culicoides huaynacapaci Felippe-Bauer, is described and illustrated based on female and male specimens collected biting humans in Department of Cajamarca, in Peruvian Amazonia. The new species is compared with its similar congener C. albuquerquei Wirth & Blanton.Uma nova espécie de Culicoides do subgênero Mataemyia Vargas, Culicoides huaynacapaci Felippe-Bauer, é descrita e ilustrada baseada em exemplares fêmeas e machos coletados picando humanos no Estado de Cajamarca, na Amazônia Peruana. A nova espécie é comparada com a espécie afim, C. albuquerquei Wirth & Blanton.

Bartonellosis and rickettsiosis are commonly reported in Peru. In order to detect Bartonella sp. and Rickettsia sp. in fleas, ticks and lice, specimens from five distinct locations in Peru (Marizagua, Cajaruro, Jamalca, Lonya Grande and El Milagro) were collected and screened for the presence of these bacteria using PCR and later confirmation by DNA sequencing. The specimens collected were distributed in 102 pools (76 Ctenocephalides felis, 2 Ctenocephalides canis, 16 Pulex irritans, 5 Pediculus humanus, 2 Rhiphicephalus sanguineus, and 1 Boophilus spp.), where Bartonella was detected in 17 pools (6 of C. felis, 9 of P. irritans, 1 of C. canis, and 1 P. humanus). Also, Rickettsia was detected in 76 pools (62 C. felis, 10 P. irritans, 2 P. humanus, and 2 C. canis). Bartonella clarridgeiae was detected in C. felis, C. canis and P. irritans pools at 5.3%, 50% and 12.5%, respectively. Bartonella rochalimae was detected in one C. felis and two P. irritans pools at 1.3% and 12.5%, respectively. Furthermore, B. henselae was detected in one C. felis pool and one P. humanus pool corresponding to 1.3% and 20%, respectively; and Bartonella spp. was also found in 5 pools of P. irritans at 31.3%. Additionally, R. felis was detected in C. felis, C. canis and P. irritans pools at 76.3%, 100% and 37.5%, respectively; and Rickettsia spp. was detected in C. felis, P. irritans and P. humanus pools at 5.3%, 25% and 40%, respectively. These results demonstrate the circulation of these bacteria in Peru.