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Introducción: La leishmaniasis cutánea (LC) es una enfermedad zoonótica endémica en Panamá. Su agente causal son protozoarios del género Leishmania y la transmiten insectos flebotominos. Objetivo: Evaluar los factores de riesgos asociados con la LC y la diversidad de flebotominos en dos comunidades rurales de Panamá Oeste. Metodología: Se seleccionaron dos comunidades endémicas para LC: Trinidad de las Minas (TM), de alta incidencia y Las Pavas (LP), de baja incidencia. Los factores de riesgo asociados con la LC fueron evaluados mediante una encuesta aplicada a100 personas (TM: n=50; LP: n=50). Se colectaron flebotominos con trampas CDC durante tres noches consecutivas en temporada lluviosa y seca. Resultados: La mayoría de las personas confirmó conocer sobre la LC (TM: 96% y LP: 68%). No se encontraron diferencias significativas entre las características sociodemográficas, estructura de las viviendas, composición del peridomicilio y abundancia/diversidad de animales domésticos en ambas comunidades. El reporte de perezosos cercanos al peridomicilio fue mayor en TM (70%) vs LP (32%). La especie de flebotomino antropofílica más abundante durante la temporada seca fue Lutzomyia gomezi (TM: 40.1% y LP: 10.4%). Durante la temporada lluviosa fue Nyssomyia trapidoi (43.4%) en TM y Psychodopygus panamensis (75.7%) en LP. Las especies zoofílicas más comunes en ambas comunidades fueron Trichopygomyia triramula y Pressatia dysponeta. Conclusión: La mayor incidencia de LC en TM podría estar condicionada a su ecología montañosa, con una cobertura boscosa cercana más extensa y una mayor frecuencia de mamíferos reservorios silvestres. Se confirmó la presencia de vectores de LC en el peridomicilio de ambas comunidades.

Didelphis marsupialis has been reported as a competent reservoir for trypanosomatid parasites infections. The aim of this study was to measure Trypanosoma cruzi, T. rangeli, and Leishmania spp. infection rates and to characterize discrete typing units (DTUs) of T. cruzi in D. marsupialis from two Chagas disease endemic sites in Panama. Blood from 57 wild-caught D. marsupialis were examined from two rural communities, Las Pavas (N = 18) and Trinidad de las Minas (N = 39). Twenty-two (38.60%) opossums were positive for flagellates by general hemoculture. T. cruzi infection was confirmed by positive hemoculture and/or kDNA based PCR performed in 31/57 (54.39%) blood samples from opossums. T. rangeli infection was confirmed by hemoculture and/or TrF/R2-Primer PCR assay applied on 12/57 (21.05%) blood samples. Nine (15.79%) D. marsupialis harbored T. cruzi/T. rangeli coinfections. All opossums tested negative for Leishmania spp. by PCR assays based on kDNA and HSP70 gene amplification. There was a significant association between T. cruzi infection and site (Fisher exact test, p = 0.02), with a higher proportion of T. cruzi infected opossums in Las Pavas (77.78%, n = 14/18) compared to Trinidad de las Minas (43.59%, n = 17/39). A significant association was found between habitat type and T. cruzi infection in opossums across both communities, (X2 = 6.91, p = 0.01, df = 1), with a higher proportion of T. cruzi infection in opossums captured in forest remnants (76%, 19/25) compared to peridomestic areas (37.5%, 12/32). T. rangeli detection, but not T. cruzi detection, may be improved by culture followed by PCR. TcI was the only DTU detected in 22 T. cruzi samples using conventional and real-time PCR. Eight T. rangeli positive samples were characterized as KP1(-)/lineage C. Trypanosome infection data from this common synanthropic mammal provides important information for improved surveillance and management of Chagas disease in endemic regions of Panama., Graphical abstract Image 1, Highlights • Trypanosoma cruzi infection is common in Didelphis marsupialis from the studied sites. • T. rangeli infection was confirmed in many opossums. • All opossums tested negative for Leishmania infection. • A higher proportion of T. cruzi infected opossums came from forest remnants. • T. cruzi parasites were characterized as TcI and T. rangeli as KP1(-)/lineage C.

Blastocystis sp. is a commonly reported intestinal parasite with a worldwide distribution. Phylogenetic analyses describe at least 17 subtypes for this parasite, and nine of them have been found in humans. However, the prevalence and some epidemiological characteristics of this parasitic infection in rural communities are not well known. The objective of this cross-sectional study was to evaluate the prevalence, subtypes, and epidemiological factors related to Blastocystis sp. Infection in children from of a small rural community in the central area of Panama. For this, 66 fecal samples from children (1 to 12 years old), were initially analyzed for the presence of parasites by a formalin-ethyl acetate/concentration method. Molecular detection and identification of Blastocystis sp. subtypes were carried out by amplification and sequencing of a partial fragment of the small-subunit ribosomal RNA gene. Using data from a questionnaire, analyses of epidemiological conditions potentially associated with Blastocystis sp. transmission were also conducted. Microscopic diagnostics showed that 33.3% (22/66) of the analyzed samples presented entero-parasites. Among them, Blastocystis sp. was the most prevalent, with 21.2% (14/66), followed by the E. histolytica/dispar/moshkovskii complex 4.5% (3/66), Giardia lamblia 1.5% (1/66) and Strongyloides stercoralis 1.5% (1/66). PCR-based analyses detected a prevalence of Blastocystis sp. infection of 74.2% (49/66) in apparently healthy children. Phylogenetic analysis revealed two different subtypes of this parasite: ST1 with 42.2% (28/66) infected, and ST3 with 31.8% (21/66) infected. In addition, recent diarrhea was significantly associated with Blastocystis sp. infection. None of the other risk factors evaluated was statistically associated with infection. These results highlight the need to further investigate clinical, epidemiological, and genetic characteristics of Blastocystis sp. infections in this community., Highlights • Infection with Blastocystis sp. is very frequent among children from a rural community in Panama • ST1 and ST3 subtypes were found with a slight predominance of ST1 • Statistical analysis showed that recent diarrhea in children was significantly related to Blastocytis sp. infection • Additional clinical and epidemiological studies related to blastocystosis in this community are needed

American cutaneous leishmaniasis (ACL) is a common and important vector-borne parasitic zoonosis in Panam ´a. Here, we study Leishmania spp. infection rates and blood-feeding patterns among common sand flies in Trinidad de Las Minas, a rural community with hyperendemic ACL transmission, and where a deltamethrin fogging trial was performed. Sand flies were collected from April 2010 to June 2011 with light traps installed inside and in the peridomicile of 24 houses. We restricted our analysis to the most abundant species at the study site: Lutzomyia trapidoi, Lutzomyia gomezi, Lutzomyia panamensis, Lutzomyia triramula, and Lutzomyia dysponeta. We detected Leishmania spp. infection in sand flies by polymerase chain reaction (PCR) amplification of the internal transcribed spacer region 1 (ITS-1) in pooled females (1–10 females per pool). Host species of engorged sand flies were identified using a cytochrome b PCR. From 455 sand fly pools analyzed, 255 pools were positive for Leishmania spp., with an estimated infection rate (confidence interval) of 0.096 [0.080–0.115] before the deltamethrin fogging which slightly, but not significantly (P > 0.05), increased to 0.116 [0.098–0.136] after the deltamethrin fogging. Blood meal analysis suggested that pigs, goats, and birds were the most common sand fly blood sources, followed by humans and domestic dogs. DNA sequencing from a subsample of ITS-1 positive pools suggests that Leishmania panamensis, Leishmania naiffi, and other Leishmania spp. were the parasite species infecting the most common vectors at the study site. Our data confirm an association between sand fly species, humans, domestic dogs, and pigs and Leishmania spp. parasites in rural Panamá. Resumen. La leishmaniasis cutánea americana (LCA) es una zoonosis parasitaria común e importante transmitida por vectores en Panamá. Aquí estudiamos las tasas de infección por Leishmania spp. y los patrones de alimentación sanguínea entre las moscas de la arena comunes en Trinidad de Las Minas, una comunidad rural con transmisión hiperendémica de LCA, y donde se realizó un ensayo de nebulización con deltametrina. Las moscas de la arena fueron recolectadas desde abril de 2010 hasta junio de 2011 con trampas de luz instaladas dentro y en el peridomicilio de 24 casas. Restringimos nuestro análisis a las especies más abundantes en el lugar de estudio: Lutzomyia trapidoi, Lutzomyia gomezi, Lutzomyia panamensis, Lutzomyia triramula y Lutzomyia dysponeta. Se detectó la infección por Leishmania spp. en los flebótomos mediante la amplificación por reacción en cadena de la polimerasa (PCR) de la región espaciadora transcrita interna 1 (ITS-1) en hembras agrupadas (1-10 hembras por grupo). Las especies hospedadoras de las moscas de la arena engordadas se identificaron mediante una PCR del citocromo b. De los 455 grupos de moscas de la arena analizados, 255 grupos resultaron positivos a Leishmania spp., con una tasa de infección estimada (intervalo de confianza) de 0,096 [0,080-0,115] antes de la nebulización con deltametrina, que aumentó ligeramente, pero no significativamente (P > 0,05), a 0,116 [0,098-0,136] después de la nebulización con deltametrina. El análisis de la harina de sangre sugirió que los cerdos, las cabras y los pájaros eran las fuentes de sangre más comunes de la mosca de la arena, seguidos por los humanos y los perros domésticos. La secuenciación del ADN de una submuestra de grupos positivos al ITS-1 sugiere que Leishmania panamensis, Leishmania naiffi y otras Leishmania spp. fueron las especies de parásitos que infectaron a los vectores más comunes en el lugar del estudio. Nuestros datos confirman una asociación entre las especies de moscas de la arena, los seres humanos, los perros domésticos y los cerdos y los parásitos de Leishmania spp. en las zonas rurales de Panamá. Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Panamá Universidad de Panamá, Panamá Instituto Costarricense de Investigación y enseñanza en Nutrición y Salud (INCIENSA), Costa Rica Universidad Nacional, Costa Rica Escuela de Medicina Veterinaria

[This corrects the article DOI: 10.1371/journal.pone.0169047.].

46 páginas, 3 figuras, 5 tablas. -- The definitive version is available at: http://www.elsevier.com. -- Technical assistance, María González (IRNASA; CSIC), Intestinal parasitic infections are among the most common infections worldwide, leading to illness with serious and long lasting implications in children and immunocompromised people. Transmission of intestinal parasites is more frequent in tropical and sub-tropical areas where sanitation is poor and socioeconomic conditions are deficient. Panama is a country where climate and social conditions could be reflected in a high number of people infected with intestinal parasites. The presence, prevalence, and distribution of intestinal parasites in this country have been approached to date only in very restricted areas and population groups, but the impact of intestinal parasite infections at the national level is unknown. Methodology/Principal Findings. We conducted a cross-sectional survey between 2008 and 2010 to determine the prevalence of intestinal parasites across Panama. Overall, 14 municipalities in seven provinces of Panama were surveyed. The presence of eggs, cysts, and larvae was assessed by microscopy in 1,123 human fecal samples using a concentration technique. A questionnaire to identify risk factors associated with the frequency of intestinal parasites in the study population was also prepared and performed. Overall, 47.4% of human samples presented parasites. Variables including community type, age group, occupation, co-presence of commensals and socioeconomic factors (use of shoes and type of sanitation) were significantly associated with intestinal parasites (p

Ecoepidemiological scenarios for Trypanosoma cruzi Chagas transmission are partially shaped by kissing bug vector ecology. The presence of Attalea butyracea Kunth, the ‘royal palm’, is a major risk factor for Chagas disease transmission in Panamá given their frequent infestations by Rhodnius pallescens Barber, a major neotropical T. cruzi vector. It was assumed that in Panamá this relationship was very close and unique, limiting the niche of R. pallescens to that of Att. butyracea. However, here we present observations about T. cruzi-infected R. pallescens infesting coyol palms, Acrocomia aculeata Jacquin, in Pedasí district, Los Santos Province, Panamá. Between May 2015 and August 2016, we sampled kissing bugs from 83 coyol palms using mice-baited traps placed at the crown of each palm during the dry and wet season. We collected 62 R. pallescens and one Eratyrus cuspidatus Stål kissing bugs. Using logistic regression, we found that the probability of kissing bug infestation in coyol palms increased during the rainy season, with infructescence number and palm height. We examined adult R. pallescens bugs (n = 30) and found T. cruzi in 67% of the samples. We were able to isolate and characterize T. cruzi from parasites present in the feces from R. pallescens, all parasites belonging to the TC I lineage. We found that green fronds number and house proximity increased T. cruzi infection probability in kissing bugs collected in coyol palms. These results highlight coyol palms as a potential risk factor for Chagas disease transmission in the dry arch of Panamá. Los escenarios ecoepidemiológicos de la transmisión de Trypanosoma cruzi Chagas están parcialmente determinados por la ecología del vector de la chinche besucona ecología del vector. La presencia de Attalea butyracea Kunth, la "palma real", es un importante factor de riesgo para la transmisión de la enfermedad de Chagas en Panamá debido a las frecuentes infestaciones por Rhodnius pallescens Barber, un importante vector neotropical de T. cruzi. Se asumió que en Panamá esta relación era muy estrecha y única, limitando el nicho de R. pallescens al de Att. butyracea. Sin embargo, aquí presentamos observaciones sobre R. pallescens infectada por T. cruzi infestando palmas de coyol, Acrocomia aculeata Jacquin, en el distrito de Pedasí, provincia de Los Santos, Panamá. Entre mayo 2015 y agosto de 2016, muestreamos chinches besuconas de 83 palmeras coyol utilizando trampas cebadas con ratones colocadas en la corona de cada palma durante la estación seca y húmeda. Se recolectaron 62 chinches besuconas R. pallescens y una Eratyrus cuspidatus Stål besucones. Utilizando una regresión logística, encontramos que la probabilidad de infestación por chinches besuconas en las palmeras coyol aumentaba durante la estación lluviosa, con el número de infrutescencia y la altura de la palma. Examinamos chinches adultas de R. pallescens (n = 30) y encontramos T. cruzi en el 67% de las muestras. Pudimos aislar y caracterizar T. cruzi a partir de parásitos presentes en las heces de R. pallescens, todos ellos parásitos pertenecientes al linaje TC I. Encontramos que las frondas verdes y la proximidad de la casa aumentaban la probabilidad de infección por T. cruzi en las chinches de beso recogidas en las palmeras coyol. Estos resultados destacan a las palmas de coyol como un potencial factor de riesgo para la transmisión de la enfermedad de Chagas en el arco seco de Panamá. Universidad Nacional, Costa Rica Escuela de Medicina Veterinaria

Flight dispersal of the triatomine bug species Rhodnius pallescens Barber, the principal vector of Chagas disease in Panama, is an important mechanism for spreading Trypanosoma cruzi, causative agent of Chagas disease. This study measures R. pallescens flight performance using a tethered flight mill both when uninfected, and when infected with T. cruzi or Trypanosoma rangeli. Forty-four out of the 48 (91.7%) insects initiated flight across all treatments, and trypanosome infection did not significantly impact flight initiation. Insects from all treatments flew a cumulative distance ranging from 0.5 to 5 km before fatiguing. The median cumulative distance flown before insect fatigue was higher in T. cruzi- and T. rangeli-infected insects than in control insects; however, this difference was not statistically significant. There was a positive relationship between parasite load ingested and time until flight initiation in T. rangeli-infected bugs, and T. rangeli- and T. cruzi-infected females flew significantly faster than males at different time points. These novel findings allow for a better understanding of R. pallescens dispersal ability and peridomestic management strategies for the prevention of Chagas disease in Panama.