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.

[Nosocomial myiasis in Latin America and the Caribbean: an overlooked reality?]. / Miasis nosocomiales en América Latina y el Caribe: ¿una realidad ignorada?

Nosocomial myiasis is an infestation by fly larvae that occurs while a patient is hospitalized. To analyze the available information on nosocomial myiasis in Latin America and the Caribbean, a search was done for cases published in the last 52 years. Nine clinical cases were found for Brazil, Costa Rica, French Guiana, Honduras, and Jamaica. Two other publications mention 139 cases in El Salvador and some 32 in Colombia, respectively. The patients and environments described presented conditions that predispose to this type of infestation. Compulsory notification is not usually required for nosocomial myiasis in Latin America and the Caribbean, meaning that there is probably considerable underreporting. Awareness needs to be raised and registry improved of myiasis in the region to aid in adoption of better prevention measures, which will benefit patient care during hospitalization.

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.

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.

[Factors associated with incidence of dengue in Costa Rica]. / Factores asociados con la incidencia de dengue en Costa Rica.

OBJECTIVE: Determine the extent to which socioeconomic, demographic, geographic, and climate variables affected the incidence of dengue and dengue hemorrhagic fever (D/DH) in Costa Rica during the period 1999-2007. METHODS: A correlational epidemiologic study was conducted that analyzed the cumulative incidence of D/DH from 1999 to 2007 and its association with different variables in the country's 81 cantons. Information was obtained from secondary sources, and the independent variables used for the analysis were selected on the basis of their representativeness in terms of sociodemographic, environmental, and health coverage factors that affect the epidemiology of D/DH. These variables were divided into four groups of indicators: demographic, socioeconomic, housing, and climate and geographical. The data were analyzed by means of simple and multiple Poisson regressions. RESULTS: The Costa Rican cantons with a higher incidence of D/DH were located primarily near the coast, coinciding with some of the variables studied. Temperature, altitude, and the human poverty index were the most relevant variables in explaining the incidence of D/DH, while temperature was the most significant variable in the multiple analyses. CONCLUSIONS: The analyses made it possible to correlate a higher incidence of D/DH with lower-altitude cantons, higher temperature, and a high human poverty index ranking. This information is relevant as a first step toward prioritizing and optimizing actions for the prevention and control of this disease.

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.

Urban mosquito species (Diptera: Culicidae) of dengue endemic communities in the Greater Puntarenas area, Costa Rica.

Field studies were conducted to determine the mosquito species richness in the urban area of Greater Puntarenas in Costa Rica. Two cross-sectional entomological surveys were performed in seven localities of Puntarenas: one survey was performed during the wet season and the other during the dry season. The sections evaluated were determined by applying a stratified cluster sampling method using satellite imagery, and a sample of 26 cells (100 x 100m) was selected for the study. The number of cells per locality was proportional to the area of each locality. The presence of mosquito larvae and pupae in water-filled artificial and natural containers was determined in each cell. Infestation was expressed as a diversity index per type of container (Ii). Eight types of larvae were identified (Aedes aegypti, Culex quinquefasciatus, Culex interrogator, Culex nigripalpus, Culex corniger, Culex tarsalis, Limatus durhamii and Toxorhynchites theobaldi) and in two cases it was only possible to identify the genus (Culex sp. and Uranotaenia sp.). A. aegypti was the most common species followed by C. quinquefascitus. Diversity of wet environments can explain the co-occurrence of various culicid species in some localities. Although A. aegypti is the only documented disease vector in the area, C quinquefasciatus, C nigripalpus, and the other species of Culex could be considered potential vectors of other pathogens. The presence and ecology of all mosquito species should be studied to optimize surveillance and prevention of dengue and to prevent the emergence of other mosquito-transmitted diseases.

Prevalence of fur mites (Acari: Atopomelidae) in non-human primates of Costa Rica.

Parasites have been investigated for some New World primates; however, very little is known about ectoparasites and specifically fur mites. In this study, Alouatta palliata, Cebus capucinus, Saimiri oerstedii, and Ateles geoffroyi monkeys from different areas of Costa Rica were searched for fur mites. A total of 276 monkeys were evaluated, and 51 of them were positive for mites of the family Atopomelidae. Listrocarpus alouattae was identified on 22.3% of A. palliata; Listrocarpus capucinus on 12.8% of C. capucinus; and Listrocarpus costaricensis on 36.8% of S. oerstedii; No fur mites were found on A. geoffroyi. Sex was not considered a determinant of mite infestation, but prevalence was significantly higher in the Central Volcanic Mountain Range Conservation Area for L. alouattae (p=0.01) and in the Central Pacific Conservation Area for L. capucinus (p=0.002). These primate fur mites are highly host-specific. Differences in the geographical distribution may be due to monkey behavior and history, as well as to environmental conditions.

A geographical sampling method for surveys of mosquito larvae in an urban area using high-resolution satellite imagery.

Entomological surveys in urban areas are often biased by selecting houses or locations with known high vector densities. A sampling strategy was developed for Puntarenas, Costa Rica, using high-resolution satellite imagery. Grids from the Advanced Spaceborne Thermal Emission and Reflection Radiometer and a QuickBird classified land cover map were used to determine the optimal final grid area for surveys. A random sample (10% of cells) was selected, and sample suitability was assessed by comparing the mean percentage of tree cover between sample and total cells. Sample cells were used to obtain entomological data from 581 locations: 26.3% of all locations positive for mosquito larvae were not households, they contained 29.5% of mosquito-positive habitats and 16% of Aedes aegypti pupae collected. Entomological indices for Ae. aegypti (pupae per person, Breteau index, container index, location index) were slightly lower when only household data were analyzed. High-resolution satellite imagery and geographical information systems appear useful for evaluating urban sites and randomly selecting locations for accurate entomological surveys.

Seasonal profiles of Aedes aegypti (Diptera: Culicidae) larval habitats in an urban area of Costa Rica with a history of mosquito control.

Dengue is the most important arboviral disease worldwide and the principal vector-borne disease in Costa Rica. Control of Aedes aegypti populations through source reduction is still considered the most effective way of prevention and control, although it has proven ineffective or unsustainable in many areas with a history of mosquito control. In this study, seasonal profiles and productivity of Aedes aegypti were analyzed in the city of Puntarenas, Costa Rica, where vector control has been practiced for more than ten years. Households contained more than 80% of larval habitats identified, although presence of habitats was more likely in other locations like lots and streets. In the wet season, habitats in the "other" category, like appliances, small manholes, and miscellaneous containers, were the most frequent habitats observed as well as the most common and productive habitats for Ae. aegypti. In the dry season, domestic animal drinking containers were very common, although concrete washtubs contained 79% of Ae. aegypti pupae collected. Individually, non-disposable habitats were as likely or more likely to contain mosquito larvae, and large containers were more likely to harbor mosquito larvae than the small ones only in the dry season. Considering various variables in the logistic regressions, predictors for Ae. aegypti in a habitat were habitat type (p < 0.001), setting (p = 0.043), and disposability (p = 0.022) in the wet season and habitat capacity in the dry season (p = 0.025). Overall, traditional Ae. aegypti larval indices and pupal indices in Puntarenas were high enough to allow viral transmission during the wet season. In spite of continued vector control, it has not been possible to reduce vector densities below threshold levels in Puntarenas, and the habitat profiles show that non-household locations, as well as non-disposable containers, should be targeted in addition to the standard control activities.

Comparison of mosquito control programs in seven urban sites in Africa, the Middle East, and the Americas.

Mosquito control programs at seven urban sites in Kenya, Egypt, Israel, Costa Rica, and Trinidad are described and compared. Site-specific urban and disease characteristics, organizational diagrams, and strengths, weaknesses, obstacles and threats (SWOT) analysis tools are used to provide a descriptive assessment of each mosquito control program, and provide a comparison of the factors affecting mosquito abatement. The information for SWOT analysis is collected from surveys, focus-group discussions, and personal communication. SWOT analysis identified various issues affecting the efficiency and sustainability of mosquito control operations. The main outcome of our work was the description and comparison of mosquito control operations within the context of each study site's biological, social, political, management, and economic conditions. The issues identified in this study ranged from lack of inter-sector collaboration to operational issues of mosquito control efforts. A lack of sustainable funding for mosquito control was a common problem for most sites. Many unique problems were also identified, which included lack of mosquito surveillance, lack of law enforcement, and negative consequences of human behavior. Identifying common virtues and shortcomings of mosquito control operations is useful in identifying "best practices" for mosquito control operations, thus leading to better control of mosquito biting and mosquito-borne disease transmission.

'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.

Exposure of dogs to spotted fever group rickettsiae in urban sites associated with human rickettsioses in Costa Rica.

The zoonotic transmission cycles of Rickettsia rickettsii and other spotted fever group (SFG) rickettsiae in Latin America have usually been associated with rural or sylvatic environments, although domestic dogs can be implicated in more populated settings. In this study, exposure of dogs to SFG rickettsiae in the Greater Metropolitan Area of Costa Rica was investigated. Dogs from sites associated with human cases and from dog shelters were evaluated by indirect immunofluorescence assay (IFA) using antigen of SFG rickettsiae. Rickettsia spp. were detected in ectoparasites by polymerase chain reaction (PCR). A total 18.5% (31/168) of dogs associated with human cases and 6.8% (11/161) of dogs in shelters had IgG end titers≥64 to Rickettsia spp. The odds of being seropositive were greater in dogs from areas associated with human cases when compared to shelters (OR: 3.2; 95% C.I: 1.5-5.6). Rhipicephalus sanguineus sensu lato (s. l.) was present in all sites associated with human cases. Rickettsia felis URRWXCal2 and R. felis-like RF2125 were detected in Ctenocephalides felis, and Rickettsia sp. IbR/CRC in Ixodes boliviensis. Results demonstrate that dogs from the main urban center of Costa Rica have been exposed to SFG rickettsiae, especially in areas with known human infection. Both human and animal health sectors must be aware of possible rickettsial diseases in urban areas, where dogs may also serve as sentinels for human infection.

Detection of rickettsiae in fleas and ticks from areas of Costa Rica with history of spotted fever group rickettsioses.

Outbreaks of spotted fevers have been reported in Costa Rica since the 1950s, although vectors responsible for transmission to humans have not been directly identified. In this study, species of Rickettsia were detected in ectoparasites from Costa Rica, mostly from five study sites where cases of spotted fevers have been reported. Ticks and fleas were collected using drag cloths or directly from domestic and wild animals and pooled according to species, host, and location. Pools were analyzed initially by PCR to detect a fragment of Rickettsia spp. specific gltA gene, and those positive were confirmed by detection of htrA and/or ompA gene fragments. Partial sequences of the gltA gene were obtained, as well as at least one ompA and/or ompB partial sequence of each species. Rickettsia spp. were confirmed in 119 of 497 (23.9%) pools of ticks and fleas analyzed. Rickettsia rickettsii was identified in one nymph of Amblyomma mixtum and one nymph of Amblyomma varium. Other rickettsiae present were 'Candidatus Rickettsia amblyommii' in A. mixtum, Amblyomma ovale, Dermacentor nitens, and Rhipicephalus sanguineus s. l.; Rickettsia bellii in Amblyomma sabanerae; Rickettsia felis in Ctenocephalides felis; and Rickettsia sp. similar to 'Candidatus R. asemboensis' in C. felis, Pulex simulans, A. ovale, and Rhipicephalus microplus. Results show the presence of rickettsiae in vectors that may be responsible for transmission to humans in Costa Rica, and evidence suggests exposure to rickettsial organisms in the human environment may be common. This is the first study to report R. rickettsii in A. varium and in A. mixtum in Costa Rica.

Pathogenic potential of a Costa Rican strain of 'Candidatus Rickettsia amblyommii' in guinea pigs (Cavia porcellus) and protective immunity against Rickettsia rickettsii.

'Candidatus Rickettsia amblyommii' is a spotted fever group rickettsia that is not considered pathogenic, although there is serologic evidence of possible infection in animals and humans. The aim of this study was to evaluate the pathogenic potential of a Costa Rican strain of 'Candidatus R. amblyommii' in guinea pigs and determine its capacity to generate protective immunity against a subsequent infection with a local strain of Rickettsia rickettsii isolated from a human case. Six guinea pigs were inoculated with 'Candidatus R. amblyommii' strain 9-CC-3-1 and two controls with cell culture medium. Health status was evaluated, and necropsies were executed at days 2, 4, and 13. Blood and tissues were processed by PCR to detect the gltA gene, and end titers of anti-'Candidatus R. amblyommii' IgG were determined by indirect immunofluorescence. To evaluate protective immunity, another 5 guinea pigs were infected with 'Candidatus R. amblyommii' (IGPs). After 4 weeks, these 5 IGPs and 3 controls (CGPs) were inoculated with pathogenic R. rickettsii. Clinical signs and titers of anti-Rickettsia IgG were determined. IgG titers reached 1:512 at day 13 post-infection with 'Candidatus R. amblyommii'. On day 2 after inoculation, two guinea pigs had enlarged testicles and 'Candidatus R. amblyommii' DNA was detected in testicles. Histopathology confirmed piogranulomatous orchitis with perivascular inflammatory infiltrate in the epididymis. In the protective immunity assay, anti-Rickettsia IgG end titers after R. rickettsii infection were lower in IGPs than in CGPs. IGPs exhibited only transient fever, while CGP showed signs of severe disease and mortality. R. rickettsii was detected in testicles and blood of CGPs. Results show that the strain 9-CC-3-1 of 'Candidatus R. amblyommii' was able to generate pathology and an antibody response in guinea pigs. Moreover, its capacity to generate protective immunity against R. rickettsii may modulate the epidemiology and severity of Rocky Mountain spotted fever in areas where both species circulate.

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.

Detection of an undescribed Rickettsia sp. in Ixodes boliviensis from Costa Rica.

Ixodes boliviensis is a tick of carnivores that is common on domestic dogs. The only Rickettsia that has been detected previously in this species is 'Candidatus Rickettsia andeanae'. We report the detection of an undescribed Rickettsia sp., named strain IbR/CRC, in I. boliviensis collected from dogs in Costa Rica. Analyses of gltA, ompA, and htrA partial sequences place Rickettsia sp. strain IbR/CRC in the group of R. monacensis, also close to an endosymbiont of Ixodes scapularis and other undescribed rickettsiae. It was not possible to isolate Rickettsia sp. strain IbR/CRC in Vero E6 or C6/36 cell lines. Isolation and further characterization of Rickettsia sp. strain IbR/CRC and the other undescribed rickettsiae are required to determine their taxonomic status and pathogenic potential.

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.

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.