Characterization of a protein with unknown function (LinJ.30.3360) in Leishmania amazonensis and Leishmania infantum.

Leishmaniasis is a disease caused by trypanosomatid protozoa of the genus Leishmania. In the Americas, the species Leishmania amazonensis is predominantly associated with American cutaneous leishmaniasis (ACL) while L. infantum is an agent of visceral leishmaniasis (VL). The genome sequences of Leishmania spp. have shown that each genome can contain about 8000 genes encoding proteins, more than half of which have an unknown function (''hypotheticals") at the time of publication. To understand the biology and genome of the organisms, it is important to discover the function of these "hypothetical" proteins; however, few studies have focused on their characterizations. Previously, LinJ.30.3360 (a protein with unknown function) was identified as immunogenic to canine serum with VL and a good antigen to diagnose the visceral form in dogs. Here, we show that the LinJ.30.3360 protein is conserved in L. infantum, L. tarantolae, L. donovani, L. major, L. mexicana, L. braziliensis, L. panamensis, Leptomonas pyrrhocoris, and Leptomonas seymouri. It has been annotated as a MORN (Membrane Occupation and Recognition Nexus) domain protein. However, since the function of this motif is unknown, functional inferences based on the primary sequence are not possible. The protein has a folded ß-leaf secondary structure, and phosphorylation was the only post-translational modification (PTM) found using prediction approach. Experiments have shown that it is located close to the flagellar pocket and presents similar abundance in both L. amazonensis and L. infantum. Furthermore, because it is a conserved protein in trypanosomatids but not in mammals and also because of its antigenicity, LinJ.30.3360 may constitute a potential drug target and/or vaccine for leishmaniasis.

Canine visceral leishmaniasis in area with recent Leishmania transmission: prevalence, diagnosis, and molecular identification of the infecting species.

INTRODUCTION: Canine visceral leishmaniasis (CVL) is an endemic disease in Brazil, and integrated control actions have been adopted by the Brazilian Ministry of Health to control its spread. However, the transmission profile is unknown in areas with recent CVL cases, including Itaúna, located in the Brazilian state of Minas Gerais, where the present study was carried out. METHODS: A total of 2,302 dogs from 12 neighborhoods were serologically tested for canine VL using the current diagnostic protocol adopted by the Brazilian Ministry of Health. Test positivity rate (TPR) and CVL prevalence were determined for each neighborhood. The presence of Leishmania was assessed in 60 seropositive dogs which had been recommended for euthanasia. Twenty-two of them (37%) were asymptomatic, and 38 (63%) were symptomatic for CVL. Parasitological (myeloculture and smear/imprint) and molecular (PCR) methods were employed for Leishmania detection in bone marrow, spleen, mesenteric lymph nodes, and ear skin. The infecting Leishmania species was identified by DNA sequencing. RESULTS: CVL prevalence (per 1,000 dogs) varied from 0.0-166.67, depending on the neighborhood, with a mean of 68.96 (SD 51.38). Leishmania DNA was detected in at least one tissue from all seropositive dogs, with comparable TPR among tissues. Leishmania parasites were identified in most (54/60) seropositive dogs, and the infecting parasite was identified as Leishmania infantum in all of these. CONCLUSIONS: Prevalence of CVL is a contributor to the spread of visceral leishmaniasis in Itaúna.

Canine visceral leishmaniasis in area with recent Leishmania transmission: prevalence, diagnosis, and molecular identification of the infecting species

Abstract INTRODUCTION: Canine visceral leishmaniasis (CVL) is an endemic disease in Brazil, and integrated control actions have been adopted by the Brazilian Ministry of Health to control its spread. However, the transmission profile is unknown in areas with recent CVL cases, including Itaúna, located in the Brazilian state of Minas Gerais, where the present study was carried out. METHODS: A total of 2,302 dogs from 12 neighborhoods were serologically tested for canine VL using the current diagnostic protocol adopted by the Brazilian Ministry of Health. Test positivity rate (TPR) and CVL prevalence were determined for each neighborhood. The presence of Leishmania was assessed in 60 seropositive dogs which had been recommended for euthanasia. Twenty-two of them (37%) were asymptomatic, and 38 (63%) were symptomatic for CVL. Parasitological (myeloculture and smear/imprint) and molecular (PCR) methods were employed for Leishmania detection in bone marrow, spleen, mesenteric lymph nodes, and ear skin. The infecting Leishmania species was identified by DNA sequencing. RESULTS: CVL prevalence (per 1,000 dogs) varied from 0.0-166.67, depending on the neighborhood, with a mean of 68.96 (SD 51.38). Leishmania DNA was detected in at least one tissue from all seropositive dogs, with comparable TPR among tissues. Leishmania parasites were identified in most (54/60) seropositive dogs, and the infecting parasite was identified as Leishmania infantum in all of these. CONCLUSIONS: Prevalence of CVL is a contributor to the spread of visceral leishmaniasis in Itaúna.

Competence of non-human primates to transmit Leishmania infantum to the invertebrate vector Lutzomyia longipalpis.

Leishmaniasis is a zoonotic disease of worldwide relevance. Visceral leishmaniasis is endemic in Brazil, where it is caused by Leishmania infantum with Lutzomyia longipalpis being the most important invertebrate vector. Non-human primates are susceptible to L. infantum infection. However, little is known about the role of these species as reservoirs. The aim of this study was to evaluate the transmissibility potential of visceral leishmaniasis by non-human primates through xenodiagnosis using the phlebotomine Lu. longipalpis as well as to identify phlebotomine species prevalent in the area where the primates were kept in captivity, and assess infection by Leishmania in captured phlebotomine specimens. Fifty two non-human primates kept in captivity in an endemic area for leishmaniasis were subjected to xenodiagnosis. All primates were serologically tested for detection of anti-Leishmania antibodies. Additionally, an anti-Lu. longipalpis saliva ELISA was performed. Sand flies fed on all animals were tested by qPCR to identify and quantify L. infantum promastigotes. Eight of the 52 non-human primates were positive by xenodiagnosis, including three Pan troglodytes, three Leontopithecus rosalia, one Sapajus apella, and one Miopithecus talapoin, with estimated numbers of promastigotes ranging from 5.67 to 1,181.93 per µg of DNA. Positive animals had higher levels of IgG anti-Lu. longipalpis saliva when compared to negative animals, prior to xenodiagnosis. Captive non-human primates are capable of infecting Lu. longipalpis with L. infantum. Our findings also demonstrate the relevance of non-human primates as sentinels to zoonotic diseases. Several phlebotomine species, including Lu. longipalpis, have been identified in the area where the primates were maintained, but only one pool of Lutzomyia lenti was infected with L. infantum. This study has implications for public health strategies and conservation medicine.

Ecology of phlebotomine sand flies in an area of leishmaniasis occurrence in the Xakriabá Indigenous Reserve, Minas Gerais, Brazil.

INTRODUCTION: Leishmaniasis is a complex vector-borne infectious diseases caused by protozoan parasites in the genus Leishmania and spread by hematophagous phlebotomine sand flies (Diptera: Psychodidae, Phlebotominae). The aim of this study was to investigate the phlebotomine fauna, endophily and exophily of the species found, and possible influence of climatic factors on their populations. METHODS: The study was conducted in the Xakriabá Indigenous Reserve (XIR) in the municipality of São João das Missões in northern Minas Gerais state, Brazil. Insects were collected over three consecutive nights in the last week of each month for 12 months from July 2015 to May 2016 from four houses in four different villages. Two traps were set up in each house: one in the intra-domicile and another in the peri-domicile. RESULTS: A total of 2,012 phlebotomine sand fly specimens representing 23 species and belonging to 10 different genera were captured and identified. Among the studied villages, Riacho do Brejo showed the highest density and diversity of phlebotomine sand flies. The species Lutzomyia longipalpis (80.3%) and Nyssomyia intermedia (7.3%), which are major vectors of visceral and cutaneous leishmaniasis, respectively, had the highest population densities, both in the intra- and peri-domicile. No correlation was observed between climatic factors and the density of phlebotomine sand flies. CONCLUSIONS: The results of the present study may contribute to a better understanding and targeting of the measures for preventing and controlling leishmaniasis by the authorities responsible for indigenous health.

Ecology of phlebotomine sand flies in an area of leishmaniasis occurrence in the Xakriabá Indigenous reserve, Minas Gerais, Brazil

Abstract INTRODUCTION: Leishmaniasis is a complex vector-borne infectious diseases caused by protozoan parasites in the genus Leishmania and spread by hematophagous phlebotomine sand flies (Diptera: Psychodidae, Phlebotominae). The aim of this study was to investigate the phlebotomine fauna, endophily and exophily of the species found, and possible influence of climatic factors on their populations. METHODS: The study was conducted in the Xakriabá Indigenous Reserve (XIR) in the municipality of São João das Missões in northern Minas Gerais state, Brazil. Insects were collected over three consecutive nights in the last week of each month for 12 months from July 2015 to May 2016 from four houses in four different villages. Two traps were set up in each house: one in the intra-domicile and another in the peri-domicile. RESULTS: A total of 2,012 phlebotomine sand fly specimens representing 23 species and belonging to 10 different genera were captured and identified. Among the studied villages, Riacho do Brejo showed the highest density and diversity of phlebotomine sand flies. The species Lutzomyia longipalpis (80.3%) and Nyssomyia intermedia (7.3%), which are major vectors of visceral and cutaneous leishmaniasis, respectively, had the highest population densities, both in the intra- and peri-domicile. No correlation was observed between climatic factors and the density of phlebotomine sand flies. CONCLUSIONS: The results of the present study may contribute to a better understanding and targeting of the measures for preventing and controlling leishmaniasis by the authorities responsible for indigenous health.