Parasitic fauna of bats from Costa Rica.

Bats are important reservoirs and spreaders of pathogens, including those of zoonotic concern. Though Costa Rica hosts one of the highest bat species' diversity, no information is available about their parasites. In order to investigate the occurrence of vector-borne pathogens (VBPs) and gastrointestinal (GI) parasites of chiropterans from this neotropical area, ectoparasites (n = 231) and stools (n = 64) were collected from 113 bats sampled in Santa Cruz (site 1) and Talamanca (site 2). Mites, fleas and ticks were morphologically and molecularly identified, as well as pathogens transmitted by vectors (VBPs, i.e., Borrelia spp., Rickettsia spp., Bartonella spp.) and from feces, such as Giardia spp., Cryptosporidium spp. and Eimeria spp. were molecularly investigated. Overall, 21 bat species belonging to 15 genera and 5 families were identified of which 42.5% were infested by ectoparasites, with a higher percentage of mites (38.9%, i.e., Cameronieta sp. and Mitonyssoides sp.) followed by flies (2.6%, i.e., Joblingia sp.) and tick larvae (1.7%, i.e., Ornithodoros sp.). Rickettsia spp. was identified in one immature tick and phylogenetically clustered with two Rickettsia species of the Spotted Fever Group (i.e., R. massiliae and R. rhipicephali). The frequency of GI parasite infection was 14%, being 3.1% of bats infected by Giardia spp. (un-identified non-duodenalis species), 1.5% by Eimeria spp. and 9.4% by Cryptosporidium spp. (bat and rodent genotypes; one C. parvum-related human genotype). The wide range of ectoparasites collected coupled with the detection of Rickettsia sp., Giardia and Cryptosporidium in bats from Costa Rica highlight the role these mammals may play as spreaders of pathogens and the need to further investigate the pathogenic potential of these parasites.

Anopheles albimanus (Diptera: Culicidae) Ensemble Distribution Modeling: Applications for Malaria Elimination.

In the absence of entomological information, tools for predicting Anopheles spp. presence can help evaluate the entomological risk of malaria transmission. Here, we illustrate how species distribution models (SDM) could quantify potential dominant vector species presence in malaria elimination settings. We fitted a 250 m resolution ensemble SDM for Anopheles albimanus Wiedemann. The ensemble SDM included predictions based on seven different algorithms, 110 occurrence records and 70 model projections. SDM covariates included nine environmental variables that were selected based on their importance from an original set of 28 layers that included remotely and spatially interpolated locally measured variables for the land surface of Costa Rica. Goodness of fit for the ensemble SDM was very high, with a minimum AUC of 0.79. We used the resulting ensemble SDM to evaluate differences in habitat suitability (HS) between commercial plantations and surrounding landscapes, finding a higher HS in pineapple and oil palm plantations, suggestive of An. albimanus presence, than in surrounding landscapes. The ensemble SDM suggested a low HS for An. albimanus at the presumed epicenter of malaria transmission during 2018-2019 in Costa Rica, yet this vector was likely present at the two main towns also affected by the epidemic. Our results illustrate how ensemble SDMs in malaria elimination settings can provide information that could help to improve vector surveillance and control.

Serological Positivity against Selected Flaviviruses and Alphaviruses in Free-Ranging Bats and Birds from Costa Rica Evidence Exposure to Arboviruses Seldom Reported Locally in Humans.

Arboviruses have two ecological transmission cycles: sylvatic and urban. For some, the sylvatic cycle has not been thoroughly described in America. To study the role of wildlife in a putative sylvatic cycle, we sampled free-ranging bats and birds in two arbovirus endemic locations and analyzed them using molecular, serological, and histological methods. No current infection was detected, and no significant arbovirus-associated histological changes were observed. Neutralizing antibodies were detected against selected arboviruses. In bats, positivity in 34.95% for DENV-1, 16.26% for DENV-2, 5.69% for DENV-3, 4.87% for DENV-4, 2.43% for WNV, 4.87% for SLEV, 0.81% for YFV, 7.31% for EEEV, and 0.81% for VEEV was found. Antibodies against ZIKV were not detected. In birds, PRNT results were positive against WNV in 0.80%, SLEV in 5.64%, EEEV in 8.4%, and VEEV in 5.63%. An additional retrospective PRNT analysis was performed using bat samples from three additional DENV endemic sites resulting in a 3.27% prevalence for WNV and 1.63% for SLEV. Interestingly, one sample resulted unequivocally WNV positive confirmed by serum titration. These results suggest that free-ranging bats and birds are exposed to not currently reported hyperendemic-human infecting Flavivirus and Alphavirus; however, their role as reservoirs or hosts is still undetermined.

Modeling the association between Aedes aegypti ovitrap egg counts, multi-scale remotely sensed environmental data and arboviral cases at Puntarenas, Costa Rica (2017-2018).

Problems with vector surveillance are a major barrier for the effective control of vector-borne disease transmission through Latin America. Here, we present results from a 80-week longitudinal study where Aedes aegypti (L.) (Diptera: Culicidae) ovitraps were monitored weekly at 92 locations in Puntarenas, a coastal city in Costa Rica with syndemic Zika, chikungunya and dengue transmission. We used separate models to investigate the association of either Ae. aegypti-borne arboviral cases or Ae. aegypti egg counts with remotely sensed environmental variables. We also evaluated whether Ae. aegypti-borne arboviral cases were associated with Ae. aegypti egg counts. Using cross-correlation and time series modeling, we found that arboviral cases were not significantly associated with Ae. aegypti egg counts. Through model selection we found that cases had a non-linear response to multi-scale (1-km and 30-m resolution) measurements of temperature standard deviation (SD) with a lag of up to 4 weeks, while simultaneously increasing with finely-grained NDVI (30-m resolution). Meanwhile, median ovitrap Ae. aegypti egg counts increased, and respectively decreased, with temperature SD (1-km resolution) and EVI (30-m resolution) with a lag of 6 weeks. A synchrony analysis showed that egg counts had a travelling wave pattern, with synchrony showing cyclic changes with distance, a pattern not observed in remotely sensed data with 30-m and 10-m resolution. Spatially, using generalized additive models, we found that eggs were more abundant at locations with higher temperatures and where EVI was leptokurtic during the study period. Our results suggest that, in Puntarenas, remotely sensed environmental variables are associated with both Ae. aegypti-borne arbovirus transmission and Ae. aegypti egg counts from ovitraps.

Mosquito (Diptera: Culicidae) Species Composition in Ovitraps From a Mesoamerican Tropical Montane Cloud Forest.

Knowledge about mosquito species diversity at tropical montane cloud forests (TMCFs) in Mesoamerica is scarce. Here, we present data on mosquito species richness from samples biweekly collected, from January to December 2017, in ovitraps installed in a TMCF patch at Vázquez de Coronado County, Costa Rica. Ovitraps were placed at 2.25, 1.50 and 0.75 m at 16 sampling points. During the study period we measured relative humidity and air temperature at each sampling point, and water temperature, volume and pH in each ovitrap. We collected a total of 431 mosquito larvae belonging to five taxonomic units, one identified to the genus level and four to the species level. The most common mosquito species was Culex bihaicolus Dyar & Nuñez Tovar (Diptera: Culicidae), which accounted for nearly 80% (n = 344) of the collected mosquitoes. Culex nigripalpus Theobald (Diptera: Culicidae) was the only medically important species we found and it was collected both in the dry (January to March) and rainy season (April to December). Over 95% (n = 411) of the mosquitoes were collected during the rainy season and 60% (n = 257) at 0.75 m. Among the environmental variables that we measured, only water volume and pH were significantly (P < 0.05) different between the dry and rainy season, the former increasing and the later decreasing during the rainy season. These results suggest that rainfall plays a major role regulating the phenology of the sampled mosquito species and highlight the need to screen for pathogens in Cx. nigripalpus at the study area.

Mosquito Species (Diptera: Culicidae) Diversity from Ovitraps in a Mesoamerican Tropical Rainforest.

Mosquito sampling using efficient traps that can assess species diversity and/or presence of dominant vectors is important for understanding the entomological risk of mosquito-borne disease transmission. Here, we present results from a survey of mosquito species sampled with ovitraps in a neotropical rainforest of Costa Rica. We found the method to be an efficient sampling tool. With a total sampling effort of 29 traps, we collected 157 fourth-instar larvae and three pupae belonging to eight mosquito taxonomic units (seven species and individuals from a homogenous taxonomic unit identified to the genus level). In our samples, we found two medically important species, Sabethes chloropterus (Humboldt) and Trichoprosopon digitatum (Rondani). The former is a proven vector of Yellow Fever in sylvatic environments and the later has been found infected with several arboviruses. We also found that mosquito species abundance and diversity increased with canopy cover and in environments where leaf litter dominated the ground cover. Finally, our results suggest that ovitraps have a great potential for systematic sampling in longitudinal and cross-sectional ecological "semi-field" studies in neotropical settings.

Análisis cromatográfico del contenido enzimático del veneno de tres serpientes peruanas / Chromatography analysis of enzyme content in three Peruvian snake venom

Las serpientes que habitan la selva amazónica tienen venenos cuya composición varía con la especie. Pero existen también especies de coste y otras que son arborícolas cuyo estudio es motivo de interés para este laboratorio. En dos serpientes de costa Bothrops pictus y Bothrops rodengheri así como en una de selva, arborícola, Bothriopsis oligolepis; se ha realizado una exploración de las principales enzimas contenidas en sus venenos luego de efectuar un fraccionamiento cromatográfico. Para ello, muestras de 50 mg. De cada veneno se fraccionaron en una columna de Sephadex G-100 con buffer acetato de amonio 0.05 M, pH 7.0 analizándose luego las actividades enzimáticas de Hialuronidasa, fosfodiesterasa, 5’ nucleotidasa, Similar a Trombina, Proteasa y Fosfolipasa A2. Los resultados mostraron variaciones muy notables en los pisos de proteínas de los venenos utilizados, así como la posición relativa y la actividad de las enzimas en estudio. En el veneno de B. oligolepis se encontraron 3 picos proteicos definidos mientras que las ponzoñas de las serpientes de costa tuvieron perfiles semejantes entre sí. Así mismo las actividades de Fosfodiesterasa, 5’ Nucleotidasa y Hialuronidasa tienen los valores de Ve/ Vo más pequeños, habiéndose encontrado en B. rodengheri esta última actividad, en cambio la Fosfolipasa A2 mostró valores de Ve/Vo notablemente elevados. También se encontraron diferencias en las actividades específicas de las enzimas investigadas pudiéndose deducir a partir de ellas, el rol biológico que cumplirían durante en envenenamiento.

In the Amazonic jungle habit snakes and their venoms have different chemical composition according to species. Besides there are some snakes from coast and others habit on trees having their venoms a particular interest to our laboratory. The venoms of Bothrops pictus and Bothrops rodengheri (both snakes) as well as Bothriopsis oligolepis (tree snake) have been in investigated on enzymatical composition after a chromatographical fractionation. 50 mg of each venom were fractionated on a Sephadex G-100 column, using 0.05 M ammonium acetate pH 7.0 as eluting. Thus, Hyaluronidase, Phosphodisterase, 5’-nucleotidase, Thrombin-like Enzyme, Proteolytic and Phospholipase A2 activities were measured after that. The results showed remarkable variation on peaks of protein obtained, as well as chromatographical position and activity in each case. On B. oligolepis venom three peaks were obtained while protein profile of coast snake venoms were similar. Phosphodiesterase, 5’-nucleotidas and Hyaluronidase exhibited little Ve/Vo values, however Hyaluronidase activity was not registered in B. rodengheri. Phospholipase A showed a great V/V values in all venoms in study. In addition specific activity differences were found among the three venoms, thus a biological effect would be related to these activity values.