Genetic variability highlights the invasion route of the Lutzomyia longipalpis complex, the main vector of Visceral Leishmaniasis in Uruguay.

In the Americas, the sandfly Lutzomyia longipalpis is the main vector of the parasitic protozoa Leishmania infantum, the etiological agent of visceral leishmaniasis (VL). The Lu. longipalpis species complex is currently discontinuously distributed across the Neotropical region, from Mexico to the north of Argentina and Uruguay. During its continental spreading, it must have adapted to several biomes and temperature amplitudes, when founder events should have contributed to the high genetic divergence and geographical structure currently observed, reinforcing the speciation process. The first report of Lu. longipalpis in Uruguay was in 2010, calling the attention of Public Health authorities. Five years later, the parasite Le. infantum was recorded and in 2015 the first case of VL in canids was reported. Hitherto seven human cases by VL have been reported in Uruguay. Here, we publish the first DNA sequences from the mitochondrial genes ND4 and CYTB of Lu. longipalpis collected in Uruguay, and we used these molecular markers to investigate their genetic variability and population structure. We described four new ND4 haplotypes in a total of 98 (4/98) and one CYTB in a total of 77 (1/77). As expected, we were able to establish that the Lu. longipalpis collected in two localities (i.e. Salto and Bella Unión) from the north of Uruguay are closely related to the populations from neighbouring countries. We also propose that the possible route for the vector arrival to the region may have been through vegetation and forest corridors of the Uruguay River system, as well as it may have benefited from landscape modifications generated by commercial forestation. The ecological-scale processes shaping Lu. longipalpis populations, the identification of genetically homogeneous groups and the gene flow among them must be carefully investigated by using highly sensible molecular markers (i.e. genome wide SNPs) since it will help to the understanding of VL transmission and contribute to the planification of public policies on its control.

Trypanosoma cruzi Isolates Naturally Adapted to Congenital Transmission Display a Unique Strategy of Transplacental Passage.

Chagas disease is mainly transmitted by vertical transmission (VT) in nonendemic areas and in endemic areas where vector control programs have been successful. For the present study, we isolated natural Trypanosoma cruzi strains vertically transmitted through three generations and proceeded to study their molecular mechanism of VT using mice. No parasitemia was detected in immunocompetent mice, but the parasites were able to induce an immune response and colonize different organs. VT experiments revealed that infection with different strains did not affect mating, pregnancy, or resorption, but despite low parasitemia, VT strains reached the placenta and resulted in higher vertical transmission rates than strains of either moderate or high virulence. While the virulent strain modulated more than 2,500 placental genes, VT strains modulated 150, and only 29 genes are shared between them. VT strains downregulated genes associated with cell division and replication and upregulated immunomodulatory genes, leading to anti-inflammatory responses and tolerance. The virulent strain stimulated a strong proinflammatory immune response, and this molecular footprint correlated with histopathological analyses. We describe a unique placental response regarding the passage of T. cruzi VT isolates across the maternal-fetal interphase, challenging the current knowledge derived mainly from studies of laboratory-adapted or highly virulent strains. IMPORTANCE The main findings of this study are that we determined that there are Trypanosoma cruzi strains adapted to transplacental transmission and completely different from the commonly used laboratory reference strains. This implies a specific strategy for the vertical transmission of Chagas disease. It is impressive that the strains specialized for vertical transmission modify the gene expression of the placenta in a totally different way than the reference strains. In addition, we describe isolates of T. cruzi that cannot be transmitted transplacentally. Taken together, these results open up new insights into the molecular mechanisms of this insect vector-independent transmission form.

Molecular detection of coccidian Apicomplexa Parasites isolated from wild crab-eating and pampas foxes through novel TaqMan™ probes: a contribution to their molecular epidemiology.

Neospora caninum, Toxoplasma gondii and Hammondia spp. are coccidian parasites similar in morphology. Molecular techniques are necessary to detect parasite DNA isolated from stool samples in wild canids because they were reported as definitive hosts of N. caninum life cycle. The objective of this study was to develop a highly sensitive and accurate molecular method for the identification of coccidian Apicomplexa parasites in crab-eating fox (Cerdocyon thous) and pampas fox (Lycalopex gymnocercus). Tissue samples from road-killed animals (pampas fox = 46, crab-eating fox = 55) and feces (pampas fox = 84, crab-eating fox = 2) were collected, and species were diagnosed through molecular assay. PCR was used for the amplification of a fragment of the coccidian Apicomplexa nss-rRNA gene. Additionally, we developed a novel real-time PCR TaqMan™ probe approach to detect T. gondii- Hammondia spp. and N. caninum. This is the first report of N. caninum DNA in pampas fox feces (n = 1), thus it was also detected from pampas fox tissues (n = 1). Meanwhile, T. gondii was found in tissues of pampas (n = 1) and crab-eating (n = 1) foxes and H. triffittae in one crab-eating fox tissue. Despite the low percentage (2.5%) of positive samples, the molecular method developed in this study proved to be highly sensitive and accurate allowing to conduct an extensive monitoring analysis for these parasites in wildlife.

Enteroparasitosis en niños de dos Centros de Atención a la Infancia y la Familia (CAIF) del barrio Casavalle, Montevideo / Children enteroparasitosis in Centers for Child and Family Care (CAIF) in Casavalle neighborhood, Montevideo

Resumen Las enteroparasitosis son enfermedades del tubo digestivo causadas por protozoarios, hongos y/o helmintos. Afectan principalmente a niños con condiciones socioeconómicas desfavorables, pudiendo tener repercusión sobre el crecimiento y desarrollo. Con el objetivo de contribuir a generar conocimiento sobre la prevalencia de las enteroparasitosis en niños de dos Centros de Atención a la Infancia y la Familia (CAIF) del barrio Casavalle, zona de influencia de la Policlínica Los Ángeles (Montevideo), entre marzo y diciembre de 2015, se realizaron coproparasitarios y espátulas adhesivas a niños mayores de 6 meses y menores de 5 años. Se devolvieron los resultados a las comunidades participantes, al centro de salud de referencia y a las autoridades de la zona. Se planificaron y realizaron jornadas de formación y sensibilización sobre las parasitosis, aportando información para su prevención. Los niños que tuvieron resultados positivos fueron tratados en la policlínica o derivados para su tratamiento. De los 577 niños matriculados, participaron en el proyecto 317 (54,9%), a todos los cuales se les realizaron los estudios coproparasitario y espátula adhesiva. Resultaron positivos 97 niños (30,6%), de los cuales 22 poliparasitados (6,9%). Los agentes más frecuentemente encontrados fueron: Giardia lamblia (18,6%), Ascaris lumbricoides (6,6%) y Enterobius vermicularis (5,7 %). Se evidenciaron tres mecanismos de transmisión con vía de entrada oral: por contaminación fecal, por contacto con el suelo y transmisión directa, por lo que son necesarios esfuerzos para mejorar hábitos de higiene personal, vigilancia epidemiológica con georreferenciación y acciones dirigidas a mejorar la salud ambiental y saneamiento.

Summary Enteroparasitosis are diseases of the digestive tract caused by protozoan, fungi and/or helminths. They mainly affect children under unfavorable socio-economic conditions, and they may have an impact on growth and development. Coproparasitary tests and Graham technique tests were performed to children between 6 months and five years old from March to December 2015, with the purpose of generating knowledge on the prevalence of enteroparasitosis in children seen in two Centers for Child and Family Care (CAIF) in Casavalle neighborhood, in the area of influence of the Los Angeles Polyclinics (Montevideo). The results of the tests were given to participants, the reference health care centers and the authorities in the area. Subsequently, activities geared to training and raising awareness on parasitosis were organized, providing information for prophylactic purposes. Children whose results were positive were treated in the polyclinic or referred for treatment. 577 children were registered in the institution and 317 of them (54.9%) participated in the project, all of whom underwent coproparasitary and Graham technique tests. 97 children were positive (30.6%), 22 of which were poliparasitized (6.9%). The most frequently found agents were Giardia lamblia (18.6%), Ascaris lumbricoides (6.6%) and Enterobius vermicularis (5.7%). Three mechanisms of transmission with oral entry route were evidenced: fecal contamination, contact with the soil and direct transmission, so efforts are needed to improve personal hygiene habits, epidemiological surveillance with georeferencing and actions aimed at improving environmental health and sanitation.

Bondades del láser y la miel, asociados al Heberprot-P® en el pie diabético. Presentación de caso / Goodness of laser and honey, associated with Heberprot-P® in diabetic foot. Case presentation

Introducción: la diabetes mellitus constituye hoy en día un serio problema de salud. Una de sus complicaciones más frecuentes es la úlcera del pie diabético asociada a alteraciones neurológicas, vasculares e infecciosas. El uso del Heberprot-P®, ha sido muy alentador. Cada día se buscan nuevas alternativas como la utilizada en el presente trabajo en combinación con la miel de abeja y el láser terapéutico. Método: se realiza una presentación de caso de una combinación terapéutica en el pie diabético. Se realizó una amplia actualización bibliográfica. Resultados: se logró acelerar el proceso de cicatrización, con una terapéutica y control estricto. Lo anterior favoreció a la cicatrización total de la ulcera en seis semanas. Conclusiones: se evidenció mediante los resultados alentadores, los beneficios de esta combinación terapéutica, en el caso que se presenta(AU)

Introduction: diabetes mellitus is a serious health problem today. One of its most frequent complications is diabetic foot ulcer associated with neurological, vascular and infectious alterations. The use of Heberprot-P® has been very encouraging. New alternatives are being sought every day as used in the present work in combination with bee honey and therapeutic laser. Method: a case presentation of a therapeutic combination is performed on the diabetic foot. An extensive bibliographic update was made. Results: it was possible to accelerate the healing process, with a strict therapeutic and control. This favored the total healing of the ulcer in six weeks. Conclusions: the benefits of this therapeutic combination were evidenced by the encouraging results, in the case presented(EU)