Highly abundant bacteria in the gut of Triatoma dimidiata (Hemiptera: Reduviidae) can inhibit the growth of Trypanosoma cruzi (Kinetoplastea: Trypanosomatidae).

Chagas disease, caused by the protozoan Trypanosoma cruzi, is a zoonosis primarily found in rural areas of Latin America. It is considered a neglected tropical disease, and Triatoma dimidiata is the main vector of the parasite in Central America. Despite efforts, Chagas disease continues to be a public health concern, and vector control remains a primary tool to reduce transmission. In this study, we tested the hypothesis that highly abundant bacteria in the gut of T. dimidiata inhibit the growth of T. cruzi. To achieve this, bacterial diversity in the gut of T. dimidiata specimens from Costa Rica was characterized by metabarcoding of the 16S rRNA, microbial isolation was performed, and the effect of freeze-dried supernatants of the isolates on T. cruzi was investigated. Metabarcoding showed that the most abundant genera in the gut were Corynebacterium, Tsukamurella, Brevibacterium, and Staphylococcus. Barcoding and sequences comparison confirmed that 8 of the 30 most abundant amplicon sequence variants (ASVs) were isolated, and 2 of them showed an inhibitory effect on the growth of T. cruzi epimastigotes. These bacteria correspond to isolates of Tsukamurella and Brevibacterium, which were respectively the second and sixth most abundant ASVs in the gut of T. dimidiata. Notably, only the isolate of Brevibacterium showed a significant difference in growth inhibition against epimastigotes of both T. cruzi strains tested. These findings suggest that the gut microbiota of T. dimidiata may play an active role in modulating parasite development.

Seasonal changes in the diversity, host preferences and infectivity of mosquitoes in two arbovirus-endemic regions of Costa Rica.

BACKGROUND: Mosquitoes are vectors of various arboviruses belonging to the genera Alphavirus and Flavivirus, and Costa Rica is endemic to several of them. The aim of this study was to describe and analyze the community structure of such vectors in Costa Rica. METHODS: Sampling was performed in two different coastal locations of Costa Rica with evidence of arboviral activity during rainy and dry seasons. Encephalitis vector surveillance traps, CDC female gravid traps and ovitraps were used. Detection of several arboviruses by Pan-Alpha and Pan-Flavi PCR was attempted. Blood meals were also identified. The Normalized Difference Vegetation Index (NDVI) was estimated for each area during the rainy and dry seasons. The Chao2 values for abundance and Shannon index for species diversity were also estimated. RESULTS: A total of 1802 adult mosquitoes belonging to 55 species were captured, among which Culex quinquefasciatus was the most caught species. The differences in NDVI were higher between seasons and between regions, yielding lower Chao-Sørensen similarity index values. Venezuelan equine encephalitis virus, West Nile virus and Madariaga virus were not detected at all, and dengue virus and Zika virus were detected in two separate Cx. quinquefasciatus specimens. The primary blood-meal sources were chickens (60%) and humans (27.5%). Both sampled areas were found to have different seasonal dynamics and population turnover, as reflected in the Chao2 species richness estimation values and Shannon diversity index. CONCLUSION: Seasonal patterns in mosquito community dynamics in coastal areas of Costa Rica have strong differences despite a geographical proximity. The NDVI influences mosquito diversity at the regional scale more than at the local scale. However, year-long continuous sampling is required to better understand local dynamics.

Detection of Rickettsia spp. in ticks of wildlife fauna from Costa Rica: First report of Rickettsia rhipicephali in Central America.

In the past two decades, new species of Rickettsia have been detected and described worldwide, some of them considered pathogenic for humans. Although Costa Rica is considered a biodiversity hotspot, the knowledge about rickettsiae in sylvatic ecosystems and wild animals is scarce. The aim of this preliminary study was to detect and identify species of Rickettsia in ticks collected from wild animals in Costa Rica. A total 119 ticks were collected from 16 animal host species belonging to diverse vertebrate families (Didelphidae, Procyonidae, Felidae, Choloepodidae, Bradypodidae, Myrmecophagidae, Tayassuidae, Tapiridae, Phyllostomidae, Bufonidae, Geoemydidae, Boidae, Colubridae), and they were grouped into 43 pools to detect the presence of Rickettsia spp. DNA by PCR targeting the gltA gene. In positive pools, amplicons of the ompA, sca5 (ompB), and/or htrA genes were also amplified to identify the species present. The identification of some ticks was also confirmed by molecular methods. Four species of Rickettsia were detected in eight (19%) tick pools: Rickettsia amblyommatis in four pools of Amblyomma geayi (host: Caluromys derbianus) and one pool of Amblyomma cf. parvum (host: Nasua narica), Rickettsia rhipicephali in one pool of Dermacentor latus (host: Tayassu pecari), 'Candidatus Rickettsia colombianensi' in one pool of Amblyomma sp. nymphs (host: Boa constrictor), and Rickettsia sp. genotype IbR/CRC in one pool of Ixodes cf. boliviensis (host: Puma concolor). This is the first molecular detection of R. rhipicephali in Central America, and of 'Candidatus R. colombianensi' in Costa Rica. Results show that diverse wild animals and their ticks are associated with several species of rickettsiae in Costa Rica, which may come in contact with humans and other domestic animals in sylvatic environments.

Acknowledging extraordinary women in the history of medical entomology.

Throughout history, women have been actively involved in the advancement of science, while struggling to overcome challenges to participate and a lack of recognition. Prior to 1950, most women were not included in the lists of "classical" descriptions of the iconic scientific figures nor included in the most relevant historical accounts. Since the second half of the twentieth century, great efforts have been made to recognize the contributions of women to the advancement of science, especially since formal scientific careers have been dominated by men, with limited (or no) access to women. Despite these challenging social, political and cultural contexts, many women have succeeded in making significant advancements, and their contributions are now being acknowledged. Such efforts have led to the publication of recent reviews and compilations on outstanding women in biological sciences. The field of medical entomology is inherently interdisciplinary, focusing on insects and other arthropods that affect human health, with input primarily from the biological and medical sciences and a strong public health perspective. Several reviews and book chapters describing the history of medical entomology have been published over the decades, but few women are mentioned in these publications, even though many women have contributed to this field. Much of the information on these women is currently scattered throughout the published literature and historical records on a wide range of topics, including activism, virology, vector control and even acarology. Considering that there is no single available compilation of women contributors in the history of medical entomology, this review aims to provide a list of 22 women and their contributions to this field. The list includes women from diverse backgrounds, born in the late 1800s and before 1950, who directly impacted medical entomology in various ways and in different regions of the world. This compilation is far from exhaustive, but it aims to identify role models and examples of extraordinary women to motivate the evolving future of this field.

Ticks infesting humans in Central America: A review of their relevance in public health.

Ticks are blood-sucking arthropods that parasitize most groups of terrestrial or semiaquatic vertebrates. Humans are accidental hosts to the ticks; however, in humans the ticks can cause damages varying from simple irritation to severe allergies, toxicosis, paralysis, and the transmission of pathogens, some of which can be fatal. Central America represents a narrow isthmus between North and South America and is considered a biodiversity hotspot. The importance of tick-borne diseases in this region is manifested by fatal outbreaks caused by Rickettsia rickettsii, severe and mild cases of other rickettsioses, ehrlichiosis, and tick-borne relapsing fevers, in addition to cases paralysis and strong allergic reactions. Even so, this information is scarce in most countries of this region, and there are no epidemiological data. In this article we present a review of the ticks that parasitize humans in Central America, covering data from the 19th Century to the present day. Of nearly 80 tick species reported in Central America, 28 species are reported on humans. This list includes species that thrive within homes, grazing areas and, to a lesser extent, in wild environments, both in lowland and high mountain forests. The most important genus in this region is Amblyomma, followed by Rhipicephalus and Ornithodoros, and to a lesser extent Haemaphysalis, Ixodes and Dermacentor. These data provide information on the tick species most commonly associated with humans in Central America, and highlight the potential for tick-borne diseases in wild, rural and urban regions.

Host-Feeding Patterns of the Mosquito Assemblage at Lomas Barbudal Biological Reserve, Guanacaste, Costa Rica.

Mosquito-borne pathogens have spread throughout tropical regions of the Western Hemisphere causing increased burden of disease in the region. Outbreaks of dengue fever, yellow fever, chikungunya, West Nile, and Zika have occurred over the past several years. Mosquito blood-feeding patterns need to be assayed to assist in determining which vertebrates could act as hosts of these mosquito-borne pathogens and which mosquito species could act as vectors. We conducted bloodmeal analyses of mosquitoes collected at Lomas Barbudal Biological Reserve, a dry tropical forest reserve in Costa Rica. Mosquitoes were collected using backpack aspirators and light, gravid, and resting traps, and then identified morphologically. Blood-fed mosquitoes underwent DNA extraction, PCR amplification, and sequencing of the vertebrate cytochrome b and cytochrome c oxidase 1 genes to identify vertebrate bloodmeal hosts. Several mosquitoes known to vector pathogens were found including Culex (Melanoconion) erraticus Dyar & Knab (Diptera: Culicidae), Cx. (Mel.) pedroi Sirivanakarn & Belkin, Aedes (Stegomyia) albopictus Skuse, Ae. (Ochlerotatus) scapularis Rondani, Ae. (Och.) serratus Theobald, and Ae. (Och.) taeniorhynchus Wiedemann. The most common bloodmeal hosts were basilisk lizards (Basiliscus vittatus) Wiegmann (Squamata: Corytophanidae) in Culex (Linnaeus) species and white-tailed deer (Odocoileus virginianus) Zimmermann (Artiodactyla: Cervidae) in Aedes (Meigen) species. These results show the diversity of mosquito species in a tropical dry deciduous forest and identify associations between mosquito vectors and potential pathogen reservoir hosts. Our study highlights the importance of understanding interactions between vector species and their hosts that could serve as predictors for the potential emergence or resurgence of mosquito-borne pathogens in Costa Rica.

The Impact of Deforestation, Urbanization, and Changing Land Use Patterns on the Ecology of Mosquito and Tick-Borne Diseases in Central America.

Central America is a unique geographical region that connects North and South America, enclosed by the Caribbean Sea to the East, and the Pacific Ocean to the West. This region, encompassing Belize, Costa Rica, Guatemala, El Salvador, Honduras, Panama, and Nicaragua, is highly vulnerable to the emergence or resurgence of mosquito-borne and tick-borne diseases due to a combination of key ecological and socioeconomic determinants acting together, often in a synergistic fashion. Of particular interest are the effects of land use changes, such as deforestation-driven urbanization and forest degradation, on the incidence and prevalence of these diseases, which are not well understood. In recent years, parts of Central America have experienced social and economic improvements; however, the region still faces major challenges in developing effective strategies and significant investments in public health infrastructure to prevent and control these diseases. In this article, we review the current knowledge and potential impacts of deforestation, urbanization, and other land use changes on mosquito-borne and tick-borne disease transmission in Central America and how these anthropogenic drivers could affect the risk for disease emergence and resurgence in the region. These issues are addressed in the context of other interconnected environmental and social challenges.

New records and phylogenetic position of Ornithodoros knoxjonesi (Ixodida: Argasidae).

Larvae of Ornithodoros knoxjonesi collected at five localities in three countries were studied using morphological and molecular methods to confirm this species' taxonomic validity. The larva of O. knoxjonesi is characterized as having 14 pairs of dorsal setae, eight pairs of ventral setae, plus a posteromedian seta; an elongate dorsal plate, tapered anteriorly; and a hypostome that is narrower near its midlength, with posteriorly projecting denticles. Although the larvae of O. knoxjonesi and Ornithodoros peropteryx are morphologically quite similar, the larva of O. knoxjonesi is characterized as having dorsal setae that are wider at the tip than at the base, while in O. peropteryx these setae are narrower at the tip than at the base; moreover, the dorsal setae are shorter in O. knoxjonesi (Al 0.037-0.065; Pl 0.035-0.059) than in O. peropteryx (Al 0.120-0.132; Pl 0.080-0.096). These species also differ in that O. knoxjonesi possesses only the Al seta on tarsus I, whereas O. peropteryx has both Al and Pl setae. And while both species have two setae on coxae I-III, in O. knoxjonesi the anterior seta is tapering and smooth and the posterior is fringed, while both setae are fringed in O. peropteryx. At the molecular level, based on a maximum likelihood analysis using approximately 400 bp of the mitochondrial 16S rDNA gene, O. knoxjonesi appears as an independent lineage, separated from O. peropteryx by a genetic distance of 16.28 %. Balantiopteryx plicata is a common host of O. knoxjonesi; however, in this work we report Pteronotus personatus and Pteronotus gymnonotus as new hosts of this tick species.

Cimex lectularius Linnaeus, 1758 (Hemiptera: Cimicidae) in Costa Rica: First Case Report Confirmed by Molecular Methods in Central America.

Cimex lectularius and Cimex hemipterus are the most common species of bedbugs that infest homes. Although case reports decreased substantially by the end of the 20th century, bed bugs, and especially C. lectularius, are currently suffering a resurgence mostly attributed to insecticide resistance, inadequate pest control, and increased travel. Here, we report, to the best of our knowledge, the first molecular confirmation of C. lectularius in Central America. Specimens were obtained from an apartment located in Heredia, Costa Rica. These specimens were identified morphologically as C. lectularius. The species identification was confirmed by amplifying and sequencing fragments of the cytochrome oxidase subunit I (COI) and the 16S rRNA (16S) genes. The phylogenetic analysis showed that the sequences obtained were more closely related to a C. lectularius mitochondrial complete genome sequence from China, with similarities of 98.84% (686/694) for COI and 98.97% (387/391) for 16S. The finding of C. lectularius in Costa Rica will require further investigation in order to determine the extent of current infestations and the susceptibility to insecticides, especially due to the impact that this species can have in human health, as well as the tourism industry in the region.

Exposure of dogs to Rickettsia spp. in Costa Rica: Risk factors for PCR-positive ectoparasites and seropositivity.

Infection of dogs with Rickettsia spp. can result in inapparent, mild, or severe disease. Moreover, common dog ticks and fleas are able to transmit rickettsiae to nearby humans. In this study, the seroprevalence of spotted fever group (SFG) rickettsiae was determined in dogs of Costa Rica, as well as possible risk factors associated with exposure. An interview of owners and clinical examinations were performed in a country-wide sample of 441 dogs. IgG antibodies were determined in 399 dogs by indirect immunofluorescence assay (IFA) using antigens of Rickettsia rickettsii, R. amblyommatis, and R. felis. The presence of Rickettsia spp. gltA gene was evaluated by PCR in ticks and fleas. Poisson regression was performed to assess possible risk factors associated with seropositivity, as well as with having PCR-positive ticks and fleas. The overall seroprevalence to SFG rickettsiae was 10.0% (end titers 64 to 256). Rhipicephalus sanguineus s.l. (116/441; 26.3%) and Ctenocephalides felis (153/441; 34.7%) were the most common ectoparasites. Rickettsia DNA was detected in 30% (39/130) and 32.3% (56/173) of tick and flea pools, respectively. Seropositivity was significantly associated with mean age of 2 to 7 years, scrotal edema, walking problems, large size, and tick and flea infestation. Being a purebred dog was a possible protective factor. The presence of Rickettsia PCR-positive ticks was associated with being a purebred dog, while flea treatment was protective. Having PCR-positive fleas was associated with being purebred and the number of people in the dog's environment; protective factors were free roaming and being an outdoor dog. Results confirm that dogs in Costa Rica are exposed to different species of SFG rickettsiae. This may represent a risk to human health and underscores the need for accurate diagnosis in dogs and humans. Surveillance of rickettsial infection in canines may provide useful indicators to understand the epidemiology of these zoonoses.

Papel potencial de Aedes albopictus Skuse en la transmisión de virus dengue (DENV) en una zona de actividad piñera de Costa Rica

RESUMEN Objetivo: Evaluar, bajo una perspectiva ecológica, la presencia de Aedes albopictus y su infección natural por virus dengue (DENV) en una zona de actividad piñera de Costa Rica. Método: Se colectaron mosquitos adultos en galerías forestales colindantes con piñeras, viviendas en proximidad a cultivos (<1 km) y viviendas en lejanía (110 km). Se empleó el índice de Shannon-Wiener para estimar biodiversidad. La infestación larvaria se evaluó en plantas de piña y viviendas y se calcularon índices aédicos de viviendas (IV) y de contenedores (IC). La detección de DENV en adultos (cuerpos y cabezas) y en larvas de Ae. albopictus se efectuó mediante RT-PCR y secuenciación. Resultados: Se colectaron 1376 adultos en total: Ae. albopictus (5,81%), Anopheles apicimacula (5,01%), Culex coronator (11,55%), Cx. inflictus (6,1%), Cx. nigripalpus (48,11%), Cx. quinquefasciatus (23,34%) y Limatus durhamii (0,07%). El índice de biodiversidad fue mayor en galerías forestales. Ae. albopictus adultos fueron colectados principalmente en el área de piñeras (73/80), aunque sólo dos larvas en las plantas de piña. Los índices aédicos en proximidad (IV: 40,7%, IC: 26,9%) y en lejanía (IV: 51,7%, IC: 29,6%) no mostraron diferencias significativas (IV Z=0,56, p=0,58; IC Z=0,16, p=0,87). Se detectó DENV-2 y DENV-3 en 2/20 grupos de cabezas y DENV-1 en 2/74 grupos de larvas de Ae. albopictus. Discusión: Las galerías forestales próximas a cultivos de piña podrían considerarse "islas ecológicas" adecuadas para el refugio de Ae. albopictus. La presencia de DENV en adultos y larvas sugiere un papel activo de Ae. albopictus en la transmisión de virus en este ecosistema.

ABSTRACT Objective To evaluate, under an ecological perspective, the presence of Aedes albopictus and the wild infection by dengue viruses (DENV) in an area of pineapple activity in Costa Rica. Materials and methods Adult mosquitoes were collected in forest galleries limiting pineapple plantations, houses adjacent to plantations (<1 km), and distant houses (1-10 km). Shannon-Wiener index was used to estimate biodiversity. Larval infestation was evaluated in pineapple plants and houses, and aedic house (HI) and container (CI) indices were calculated. Detection of DENV in Ae. albopictus adults (bodies and heads) and larvae was performed by RT-PCR and sequencing. Results A total 1376 adult mosquitoes were collected: Ae. albopictus (5.81%), Anopheles apicimacula (5.01%), Culex coronator (11.55%), Cx. inflictus (6.1%), Cx. nigripalpus (48.11%), Cx. quinquefasciatus (23.34%), and Limatus durhamii (0.07%). Biodiversity index was higher in forest galleries. Most adult Ae. albopictus were collected in forests close to pineapple fields (73/80), although only 2 larvae were detected in pineapple plants. Larval indices in adjacent houses (HI: 40.7%, CI: 26.9%) and distant houses (HI: 51.7%, CI: 29.6%) were similar (HI Z=0.56, p=0.58; CI Z=0.16, p=0.87). DENV-2 and DENV-3 were detected in 2/20 "pools" of Ae. albopictus heads and DENV-1 in 2/74 "pools" of larvae. Conclusion Forest galleries that are in proximity to pineapple plantations could be considered "ecological islands" that are suitable for refuge of Ae. albopictus. Presence of DENV in adults and larvae suggests an active role for Ae. albopictus in virus transmission within this ecosystem.

Determinación del intervalo post mortem mínimo (IPM) basado en un modelo de acumulación térmica con una cepa de Lucilia eximia (Diptera: Calliphoridae) de Costa Rica / Lucilia eximia (Diptera: Calliphoridae) as forensic indicator for the calculation of the post mortem interval in Costa Rica

RESUMEN Introducción: L. eximia es una especie de amplia distribución geográfica en Costa Rica, cuyas poblaciones son estables durante todo el año. Las características biológicas de ésta permiten concebirla como un potencial indicador forense para el cálculo del intervalo post mortem mínimo (IPM). Objetivo: Comparar el IPM calculado mediante un modelo de acumulación térmica con L. eximia (Diptera: Calliphoridae), con el IPM real en cadáveres de cerdos domésticos (Sus scrofa). Material y Métodos: Dos cadáveres de cerdo fueron expuestos en un bosque premontano húmedo tropical. A partir de dichos cadáveres se colectaron larvas de L. eximia a las 78, 144 y 192 horas posteriores al momento de muerte y se propició su desarrollo en condiciones controladas. Con los datos generados, se realizaron retroproyecciones térmicas para efectuar el cálculo del IPM. Resultados: L. eximia requirió en total 7 236,50 grados- hora acumulados (ADH) para culminar su desarrollo. Los IPM calculados mediante el modelo de acumulación térmica para los tres momentos de colecta fueron 112,56; 153,60 y 104,64 horas. Únicamente el valor intermedio no presentó diferencias estadísticamente significativas con el tiempo de muerte conocido (p>0.05). Conclusiones: El modelo de acumulación térmica en que se utilizó L. eximia brindó información precisa sobre el IPM sólo cuando se tomaron las muestras en el campo a las 144 horas, lo que perfila su utilidad para las primeras etapas de la descomposición cadavérica.

ABSTRACT Introduction: L. eximia is a species that shows a wide geographical distribution in Costa Rica, whose populations are stable throughout the year. The biological characteristics of L. eximia allow it to be considered as a potential forensic indicator for the calculation of the minimum post mortem interval (PMI). Objective: To compare the PMI calculated by a thermal accumulation model with L. eximia (Diptera: Calliphoridae), with the PMI in corpses of pigs (Sus scrofa). Methods: Two pig corpses were exposed in a tropical premontane moist forest. Larvae of L. eximia were collected from the pig carcasses at 78, 144 and 192 hours after the moment of death, and their development was promoted under controlled conditions. With the data generated, thermal back-projections were made to calculate the PMI. Results: L. eximia required a total of 7 236.50 accumulated degree hours (ADH) to complete its life cycle. The IPMs calculated by the thermal accumulation model were 112.56, 153.60, and 104.64 hours, respectively. Only the intermediate value did not present statistically significant differences with the known time of death (p>0.05). Conclusions: The thermal accumulation model using L. eximia as a forensic indicator provided accurate information about the PMI only when samples were collected at 144 hours, which indicates that this method may be accurate and useful during the first stages of cadaveric decomposition.

Resistencia de Aedes aegypti (diptera: culicidae) a insecticidas organofosforados y piretroides en la localidad de Orotina, Alajuela, Costa Rica / Resistance of Aedes aegypti (Diptera: Culicidae to organophosphate and pyrethroid insecticides in Orotina, Alajuela, Costa Rica

Objetivos: Determinar la resistencia de una cepa de Aedes. aegypti de Orotina a insecticidas organofosforados (temefós y malatión), y a piretroides (deltametrina, lambda cialotrina y cipermetrina). Métodos: La evaluación de la resistencia se efectuó mediante bioensayos larvarios. A partir de cada uno de los insecticidas se calculó la concentración letal 50% (CL50). También se calculó el factor de resistencia 50% (FR50) con respecto a la cepa Rockefeller, que sirvió como control susceptible. En casos de resistencia, la evaluación se repitió exponiendo previamente las larvas a butóxido de piperonilo (BP), S, S, S tributilfosforotritionato (DEF) y ácido etacrínico (AE) para establecer los mecanismos de detoxificación asociados con la resistencia. En cada caso se calculó un factor de sinergismo 50% (FS50). Resultados: La cepa Orotina mostró susceptibilidad a temefós, malatión, deltametrina y lambda cialotrina, pero mostró resistencia incipiente a cipermetrina (CL50= 0,01103 mg/L, FR50 = 5,32). Sólo el BP revertió la resistencia a este insecticida (FS50 =10,92), lo que representa un mecanismo de detoxificación asociado con el sistema citrocromo P450 monooxigenasa. Discusión: Aunque la cepa de Ae. aegyptide Orotina mostró resistencia a cipermetrina, existen otros insecticidas para los cuales fue susceptible, que brindan opciones a las autoridades de salud para su implementación en el control químico del vector. Conclusiones: El monitoreo de la resistencia es requerido para asegurar la efectividad de los insecticidas que se utilizan en el control químico.

Abstract Objectives: To determine the resistance to organophosphate (temephos and malathion) and pyrethroid (deltamethrin, lambda cyhalothrin, and cypermethrin) insecticides in a strain of Aedes aegypti from Orotina. Methods: The evaluation of the resistance was carried out by larval bioassays. Lethal concentration 50% (LC50) was calculated for each insecticide. A factor of resistance 50% (FR50) was also calculated with respect to the Rockefeller strain, which served as susceptible control. In cases of resistance, the evaluation was replicated by exposing the larvae to piperonyl butoxide (PB), S, S, S tributylphosphorotritionate (DEF), and ethacrynic acid (AE), in order to establish the detoxification mechanisms associated with the resistance. In these cases, a factor of synergism 50% (FS50) was also calculated. Results: The Orotina strain of Ae. aegyptiwas susceptible to temephos, malathion, deltamethrin and lambda cyhalothrin, but showed incipient resistance to cypermethrin (LC50 = 0.01103 mg/L, FR50 = 5.32). Only PB reversed the state of resistance (FS50 = 10.92), which suggests a detoxification mechanism associated with the citochrome P450 monooxygenase system. Discussion: Although the Ae. aegyptistrain from Orotina showed resistance to cypermethrin, it was susceptible to other insecticides, which can be used as alternative options for chemical control of the vector. Conclusions: Monitoring of resistance is required to ensure the effectiveness of insecticides used in chemical control.

Resistencia a insecticidas en cepas de Aedes aegypti (Diptera: Culicidae) de tres distritos de la Región Pacífico Central de Costa Rica / nsecticide resistance in Aedes aegypti (Diptera: Culicidae) strains from three districts of the Central Pacific Region of Costa Rica

Introducción: Aedes aegypti es vector de virus dengue, chikungunya y Zika en Costa Rica. Su alta incidencia y carencia de vacunas hacen del control vectorial, incluyendo el químico, la única alternativa para disminuir su transmisión. El uso reiterativo de insecticidas propicia su resistencia. Objetivo: Determinar la resistencia y mecanismos de detoxificación enzimática a temefós e insecticidas piretroides en cepas de Ae. aegypti de tres distritos de la Región Pacífico Central de Costa Rica. Métodos: La resistencia a temefós, deltametrina y cipermetrina fue determinada en tres cepas de Barranca, Jacó y Quepos mediante bioensayos en larvas. Para cada insecticida se calculó la concentración letal 50 por ciento (CL50) y un factor de resistencia 50 por ciento(FR50), empleando la cepa Rockefeller como control. Ante la ocurrencia de resistencia, se repitieron los bioensayos utilizando butóxido de piperonilo, S,S,S, tributilfosforotritioato y ácido etacrínico que inhiben monooxigenasas, esterasas y glutatión S transferasa, respectivamente. Resultados: Ninguna cepa mostró resistencia a temefós. Las cepas Barranca y Jacó fueron resistentes a deltametrina (FR50= 7,38; 28,23, respectivamente). La cepa Jacó mostró, adicionalmente, resistencia a cipermetrina (FR50= 7,70). La cepa Quepos no mostró resistencia a ningún piretroide. Solo la cepa Barranca mostró disminución de resistencia a deltametrina asociada al butóxido de piperonilo denotando vinculación con monooxigenasas (FR50: 10,10). Para los otros casos de resistencia, ninguno de los sinergistas disminuyó la resistencia. Conclusiones: Las larvas de Ae. aegypti de las localidades evaluadas no mostraron resistencia a temefós. Respecto a piretroides se evidenció la aparición de resistencia que posiblemente no es mediada por detoxificación enzimática(AU)

Introduction: Aedes aegypti is the vector of dengue, chikungunya, and Zika viruses in Costa Rica. The high incidence and the lack of vaccines make vector control, including chemical control, the only measure to prevent transmission. The repetitive use of insecticides may induce resistance. Objective: To determine resistance and enzymatic detoxifying mechanisms to temephos and pyrethroids insecticides in strains of Ae. aegypti from three districts of the Central Pacific Region of Costa Rica. Methods: Resistance to temephos, deltamethrin, and cypermethrin was determined in three strains of Barranca, Jacó, and Quepos by larval bioassays. In each test, the lethal concentration 50 percent (LC50) and a factor of resistance 50 percent (FR50) were calculated, using the Rockefeller strain as control. When resistance was observed, the bioassays were repeated using piperonyl butoxide, S,S,S, tributylphosphorotritioate, and ethacrynic acid, that inhibit monoxygenases, esterases, and glutathione S- transfererase, respectively. Results: None of the strains were resistant to temephos. Resistance to deltamethrin (FR50= 7.38 and FR50= 28.23, respectively) was determined in the strains from Barranca and Jacó, while resistance to cypermethrin was detected only in Jacó (FR50= 7.70). The Quepos strain was not resistant to any pyrethroid. Only the Barranca strain showed a decrease in the resistance to deltamethrin when piperonyl butoxide was used, linking the resistance to monooxygenase enzymes (FR50: 10.10). For the other cases, none of the synergists decreased the resistance. Conclusions: Larvae of Ae. aegypti from the localities evaluated were not resistant to temephos. With respect to pyrethroids, results show an emergence of resistance that may not be mediated by enzymatic detoxification(AU)

Epidemiological characterization of incident cases of Rickettsia infection in rural areas of Urabá region, Colombia.

INTRODUCTION: Most of the studies related to rickettsial infection in Colombia are cross-sectional because of the challenge in conducting prospective studies on infectious disease that may have a difficult diagnosis. Although cross-sectional studies are essential to detect people exposed to rickettsiae, they are not suited to demonstrate the recent circulation of this pathogen in areas at risk of transmission. OBJECTIVE: To characterize the epidemiology of incident cases of Spotted fever group (SFG) rickettsial infection in humans and equines from rural areas of Urabá region in Colombia where outbreaks of rickettsiae previously occurred. MATERIALS AND METHODS: A prospective study was conducted in the Alto de Mulatos and Las Changas in the Urabá region. Serum samples and socio-ecological information were collected from 597 people enrolled in 2015, and a second sample was collected from 273 people a year later. Indirect immune-fluorescence assays for detection of IgG antibody against rickettsiae were done using slides with Rickettsia rickettsii antigens. A titer ≥128 was considered positive. Incident cases were defined as (i) serological conversion of IgG titers from seronegative to seropositive or (ii) at least a four-fold increase in IgG end point titers in the second sample. RESULTS: The cumulative incidence of rickettsial infection was 6.23% (95%CI 3.67-9.78) in humans and 32.31% (21/65) of incident cases in equines. Incident cases were mostly females (82.35%), the median age of cases was 41.02 years (IQR 18.62-54.1), and 29.41% reported tick bites during the study period. Results from multivariate analysis showed that removal of ticks after working outdoors is a protective factor for rickettsial infection (RR 0.26, 95%CI 0.08-0.84) and that a higher incidence of infection occurred in people who reported fever in the last year (RR 4.26, 95%CI 1.15-9.31). CONCLUSIONS: These results showed recent circulation of SFG rickettsiae in areas where previous lethal outbreaks have been reported, supporting the implementation of preventive measures to halt rickettsial transmission in the studied communities.

A review of the genus Rickettsia in Central America.

In this paper, we present a historical review of rickettsiosis in Central America and also the most recent findings of Rickettsia in ectoparasites. All countries of Central America have records of rickettsiosis. Regarding the typhus group rickettsioses, there is clinical or serological evidence of Rickettsia prowazekii in Guatemala, Rickettsia typhi in Panama, Guatemala, and Costa Rica and unidentified species of the typhus group in El Salvador. Concerning spotted fever group rickettsiosis, there is serological evidence of infection by Rickettsia akari in Costa Rica and confirmed cases involving Rickettsia rickettsii in Panama and Costa Rica. There are also reports of spotted fever group rickettsiosis in acute patients from Guatemala, Honduras, and Nicaragua. Serological studies in Central America show reactivity of Rickettsia ambyommatis, Rickettsia bellii, Rickettsia felis, Rickettsia rhipicephali, and R. rickettsii in domestic and wild mammals. Eight species of Rickettsia have been detected in ectoparasites from Central America: R. africae (or very similar), R. amblyommatis, R. asembonensis, R. bellii, R. felis, R. parkeri, R. rhipicephali, and R. rickettsii, in addition to undescribed strains such as Atlantic Rainforest, Colombianensi, IbR/CRC, Barva, Aragaoi, and Candidatus "Rickettsia nicoyana;" the latter being the only one associated with Argasidae (Ornithodoros knoxjonesi). R. amblyommatis is the most common species in Central America, seeing as it has been reported in 10 species of ticks and one of fleas in five of the seven countries of the region. In this study, we demonstrate that the genus Rickettsia is widely distributed in Central America and that rickettsiosis could be an underestimated problem in the absence of greater diagnostic efforts in undetermined febrile cases.

Spotted fever group Rickettsiae in Ticks from Missouri.

Tick-borne rickettsioses pose a major health threat among vector-borne infections in Missouri but there are some uncertainties regarding the vector competence and range of tick species, as well as the virulence of certain bacterial species. A survey was developed and implemented to assess local healthcare practitioners' awareness of the prevalence and diagnosis of tick-borne diseases. In addition, ticks collected from rural areas of St. Louis County, Missouri, were evaluated to detect spotted fever group (SFG) rickettsiae and to determine the most common tick species present. Physician responses showed a good general awareness of tick-borne diseases and antibiotic choices but responses varied regarding length of treatment, the most common tick-borne diseases, and tick vectors. No new tick vectors were collected in the area; Rickettsia amblyommatis was the predominant SFG species, and it was detected in Amblyomma americanum, Dermacentor variabilis, andIxodes scapularis; Rickettsia montanensis was detected in D. variabilis. The high prevalence of Rickettsia amblyommatis in these ticks suggests that there is a high risk of exposure to this SFG rickettsial species to humans and that it may be providing some cross-protective immunity to R. rickettsii.

Nuevos registros de Aedes albopictus (Skuse) en cuatro localidades de Costa Rica / New records of Aedes albopictus (Skuse) in four locations of Costa Rica

Resumen Introducción Aedes albopictus (Skuse) es un vector de arbovirus ampliamente distribuido. En Costa Rica, la expansión geográfica de Ae. albopictus se ha incrementado en la última década, por lo que es importante actualizar el conocimiento sobre su distribución. Objetivo Informar sobre el hallazgo de Aedes albopictus en nuevas localidades de Costa Rica. Materiales y Métodos Se colectaron formas inmaduras de mosquitos con morfología sugestiva de Ae. albopictus en localidades de las provincias de Puntarenas (isla de Chira y Golfito), Guanacaste (Liberia), Alajuela (Upala), San José (Acosta) y Heredia (Sarapiquí). Los especímenes fueron fijados en etanol al 70%, aclarados en lactofenol e identificados con el uso de claves dicotómicas. Resultados Las especies identificadas fueron Ae. albopictus, Aedes aegypti, Haemagogus equinus y Haemagogus iridocolor-H. lucifer. Las de Ae. albopictus procedieron de los siguientes depósitos: recipiente plástico, tanque de agua, lona plástica, balde, llanta, canoa, piscina, maceta y bota de hule. Se reporta la presencia de Ae. albopictus en Liberia (Guanacaste), isla de Chira y Golfito (Puntarenas) y Upala (Alajuela). Conclusiones Se evidencia la expansión que ha experimentado Ae. albopictus en Costa Rica. Se enfatiza la necesidad de un diagnóstico microscópico certero, pues las larvas de Ae. albopictus pueden coexistir con otras especies que resultan difíciles de diferenciar, tales como Ae. aegypti y algunas especies de Haemagogus. Considerando que Ae. albopictus es un vector competente para virus dengue (DENV), chikungunya (CHIKV) y Zika (ZIKV), se recomienda investigar su papel en la transmisión de virus en estas zonas, así como el desplazamiento y/o coexistencia con especies como Ae. aegypti.

Abstract Introduction Aedes albopictus (Skuse) is a vector of arboviruses that is widely-distributed. In Costa Rica the geographical expansion of Ae. albopictus has increased in the past decade, which requires updating knowledge about its distribution. Objective To report the finding of Aedes albopictus in new localities of Costa Rica. Materials and Methods Immature stages of mosquitoes that were morphologically suggestive of Ae. albopictus were collected in localities of the provinces of Puntarenas (Chira island and Golfito), Guanacaste (Liberia), Alajuela (Upala), San José (Acosta) and Heredia (Sarapiquí). Specimens were fixed in 70 % ethanol, cleared in lactophenol, and identified using dichotomous keys. Results The species identified were: Ae. albopictus, Aedes aegypti, Haemagogus equinus, and Haemagogus iridocolor-H. Lucifer. The ones of Ae. albopictus were in the following container types: plastic container, water tank, plastic tarp, bucket, tire, roof gutter, swimming pool, flower pot and rubber boot. This is the first scientific report of Ae. albopictus in Liberia (Guanacaste), Chira island and Golfito (Puntarenas), and Upala (Alajuela). Conclusions There is evidence of the expansion of Ae. albopictus in Costa Rica. Emphasis is given as to the need for a precise microscopical diagnosis, since Ae. albopictus larvae may coexist with other species that may be difficult to differentiate, such as Ae. aegypti and some species of Haemagogus. Considering that Ae. albopictus is a competent vector for dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV) viruses, further investigation of its role in virus transmission is recommended in these areas, as well as displacement and/or coexistence with species like Ae. aegypti.

'Candidatus Rickettsia nicoyana': A novel Rickettsia species isolated from Ornithodoros knoxjonesi in Costa Rica.

Rickettsiae are intracellular bacteria commonly associated with hematophagous arthropods. Most of them have been described in hard ticks, but some have been found in soft ticks. Here we report the detection and isolation of a new Rickettsia from Ornithodoros knoxjonesi larvae collected from Balantiopteryx plicata (Emballonuridae) in Nicoya, Costa Rica. Two ticks were processed to detect Rickettsia spp. genes gltA, ompA, ompB, and htrA by PCR. Part of the macerate was also inoculated into Vero E6 and C6/36 cell lines, and cells were evaluated by Giménez stain, indirect immunofluorescence assay (IFA), and PCR. Both ticks were positive by PCR and rickettsial growth was successful in Vero E6 cells. Amplification and sequencing of near full length rrs, gltA, sca4 genes, and fragments of ompA and ompB showed that the Rickettsia sp. was different from described species. The highest homologies were with 'Candidatus Rickettsia wissemanii' and Rickettsia peacockii: 99.70% (1321/1325) with both sequences for rrs, 99.58% (1172/1177) and 99.76% (1246/1249) for gltA, 99.26% with both sequences (2948/2970 and 2957/2979) for sca4, 98.78% (485/491) and 98.39% (2069/2115) for ompA, and 98.58 (1453/1474) and 98.92% (1459/1475) for ompB; respectively. Bat blood, spleen, liver, and lung samples analyzed for Rickettsia detection were negative. Results demonstrate that the Rickettsia isolated from O. knoxjonesi is probably an undescribed species that belongs to the spotted fever group, for which 'Candidatus Rickettsia nicoyana' is proposed. Considering that B. plicata inhabits areas where contact with humans may occur and that human parasitism by Ornithodoros has been reported in the country, it will be important to continue with the characterization of this species and its pathogenic potential.

Global genetic diversity of Aedes aegypti.

Mosquitoes, especially Aedes aegypti, are becoming important models for studying invasion biology. We characterized genetic variation at 12 microsatellite loci in 79 populations of Ae. aegypti from 30 countries in six continents, and used them to infer historical and modern patterns of invasion. Our results support the two subspecies Ae. aegypti formosus and Ae. aegypti aegypti as genetically distinct units. Ae. aegypti aegypti populations outside Africa are derived from ancestral African populations and are monophyletic. The two subspecies co-occur in both East Africa (Kenya) and West Africa (Senegal). In rural/forest settings (Rabai District of Kenya), the two subspecies remain genetically distinct, whereas in urban settings, they introgress freely. Populations outside Africa are highly genetically structured likely due to a combination of recent founder effects, discrete discontinuous habitats and low migration rates. Ancestral populations in sub-Saharan Africa are less genetically structured, as are the populations in Asia. Introduction of Ae. aegypti to the New World coinciding with trans-Atlantic shipping in the 16th to 18th centuries was followed by its introduction to Asia in the late 19th century from the New World or from now extinct populations in the Mediterranean Basin. Aedes mascarensis is a genetically distinct sister species to Ae. aegypti s.l. This study provides a reference database of genetic diversity that can be used to determine the likely origin of new introductions that occur regularly for this invasive species. The genetic uniqueness of many populations and regions has important implications for attempts to control Ae. aegypti, especially for the methods using genetic modification of populations.