Ecology of sandflies (Diptera:Psychodidae) in a restricted focus of cutaneous leishmaniasis in northern Venezuela. III. Seasonal fluctuation

A one year-long study (March 1979-March 1980) was carried out at San Esteban, an endemic focus of cutaneous leishmaniasis in Northern Venezuela, with the aim of observing the seasonal fluctuation of the local phlebotomine sandflies species. The influence of climatic factors (temperature, relative humidity and rainfall) on population dynamics was analyzed in three collecting sites--a house, a peridomestic area and a sylvatic region. Among anthropophilic species, L. panamensis behaved as a wetseason species, the mean minimum relative humidity being the critical factor influencing the total number of individuals. When the population density of this fly decreased, it was successfully replaced by L. ovallesi, a dry-season species. On the other hand, seasonal variations of L. gomezi were more strongly affected by the temperature.     

Ecology of sandflies (Diptera: Psychodidae) in a restricted focus of cutaneous leishmaniasis in northern Venezuela. I. Description of the study area, catching methods and species composition

A study on the ecology of phlebotomine sandfly fauna in a restricted focus of cutaneous leishmaniasis in northern Venezuela was undertaken in order to investigate the species responsible for the transmission. The study area and catching methods for phlebotomine sandflies are described. A total of 9,061 females and 1,662 males were collected during a year-term study. 12 species of Lutzomya and 1 species of Brumptomya sp. were identified. Absolute and relative abundance and occurrence for each species were determined. The relative occurrence allowed to distinguish the common species, viz. L. panamensis, L. ovallesi, L. gomezi, L. trinidadensis, L. atroclavata, L. cayennensis, L. shannoni and L. olmeca bicolor from the rare species vis., L. punctigeniculata, L. rangeliana, L. evansi and L. dubitans. General comments on the species composition of the sandfly fauna in this locality are made.     

Ootaxonomic investigation of five Lutzomyia species (Diptera, Psychodidae) from Venezuela

The eggshell fine structure of five sand fly species from Venezuela belonging to the genus Lutzomyia (L. migonei, L. ovallesi, L. absonodonta, L. gomezi and L. panamensis) was examined by scanning electron microscopy. The chorionic sculpturing of L. migonei, L. ovallesi, L. absonodonta and L. gomezi was characterized by series of columns arranged in palisade to form sinuous ridges. In inter-ridge areas, the basal layer was covered with fibrous material. The outer chorion of L. panamensis had a pattern known as "mountain- or volcano-like". The morphology of the posterior pole and aeropyle had a common structure in the five species, with some species-specific characters. The eggshell features of the five species are compared with those of other phlebotomine sand flies.     

Lutzomyia sand flies (Diptera: Psychodidae) from middle and lower Putumayo Department, Colombia, with new records to the country

A total of 4,840 phlebotomine sand flies from 54 localities in Putumayo department (=state), in the Colombian Amazon region, were collected in Shannon traps, CDC light traps, resting places and from human baits. At least 42 Lutzomyia species were registered for the first time to the department. Psychodopygus and Nyssomyia were the subgenera with the greatest number of taxa, the most common species being L. (N.) yuilli and L. (N.) pajoti. They were sympatric in a wide zone of Putumayo, indicating that they should be treated as full species (new status). Among the anthropophilic sand flies, L. gomezi and L. yuilli were found in intradomiciliar, peridomestic, urban or forest habitats. L. richardwardi, L. claustrei, L. nocticola and L. micropyga are reported for the first time in the Colombian Amazon basin. L. pajoti, L. sipani and L. yucumensis are new records for Colombia.     

Vector competence of some Neotropical sandflies for the Leishmania (Viannia) braziliensis complex

Abstract. To evaluate the vector competence of some Lutzomyia spp. (Diptera: Psychodidae) for Leishmania (Viannia) spp. (Kinetoplastida: Trypanosoma-tidae), experimental infections of anthropophilic sandflies from the Colombian Pacific coast were performed, through membrane feeding and xenodiagnosis on hamsters infected with Le.(V.)braziliensis or Le.(V.)panamensis. Wild-caught or F, generation females of Lutzomyia gomezi, Lu.hartmanni, Lu.panamensis and Lu. trapidoi were allowed to feed on hamster lesions and then maintained at 26^oC and >80% r.h. on a sugar-water diet until dissection on the fifth day post-infection (p.i.).
Despite similar infection rates (range 37–44%) in both Lu.gomezi and Lu.trapidoi , infections were heavier (>100 parasites) in the latter species. Infections of Lu.trapidoi with Le.braziliensis ( n = 21) and Le.panamensis (n = 27) showed parasite migration toward the foregut, with promastigote colonization of the stomodeal valve and appearance of infective forms. In contrast, infections of Lu.gomezi with Le. braziliensis (n = 10) and Le.panamensis (n = 5) were light (<50 parasites) and usually restricted to the pylorus. In Lu.hartmanni , only a few promastigotes were found in the pylorus and midgut of 3/8 specimens infected with Le.braziliensis , and no Le.panamensis developed (n = 19). By day 5 p.i., promastigote colonization of the hind- and midgut by Le.panamensis was observed in 2/4 Lu.panamensis but not Le.braziliensis (n = 3).
It was concluded that Lu.trapidoi is a more efficient vector than Lu.gomezi for both Le.braziliensis and Le.panamensis , and that Lu.hartmanni and Lu.panamensis are of minor importance for Leishmania transmission in this endemic area.     

Environmental and demographic factors determining the spatial distribution of Triatoma guasayana in peridomestic and semi-sylvatic habitats of rural northwestern Argentina

Abstract.  Triatoma guasayana (Wygodzinsky & Abalos), a sylvatic vector of Chagas' disease, occurs in natural and peridomestic habitats of the dry Chaco region of Argentina, Bolivia and Paraguay. Ten-year retrospective spatial analyses of peridomestic T. guasayana abundance in the rural community of Amamá were expanded to the neighbouring community of Trinidad in northwestern Argentina. The distribution of T. guasayana in domiciles, peridomiciles (storerooms, chicken coops and corrals) and natural habitats (bromeliads, dry cacti and logs) around houses (i.e. 'semi-sylvatic' habitats) was analysed. The distribution of the 316 T. guasayana specimens collected in domestic and peridomestic sites during 1993–2002 was significantly clustered in both communities. Searches confirmed that the spatial distribution of semi-sylvatic and peridomestic T. guasayana was determined by the joint effects of the local abundance of goats and the density of semi-sylvatic habitats. The integration of detailed entomological and demographic longitudinal data with geographic information system data, high-resolution satellite imagery, appropriate spatial and temporal analyses and field observations allowed us to infer the underlying processes determining the distribution of T. guasayana in rural communities. This approach may be applied to other sylvatic and peridomestic vectors of Chagas' disease in order to identify high-risk areas for targeted control or environmental management.     

Phlebotomine sand flies (Diptera: Psychodidae) associated with the appearance of urban Leishmaniasis in the city of Sincelejo,Colombia

Although once associated only with rural areas, the American leishmaniasis vectors now appear to be associated also with urban and suburban areas of the Neotropics. Following the appearance of the first autochthonous visceral and cutaneous leishmaniasis cases in the urban area of the city of Sincelejo, Colombia, a preliminary entomological survey of the sand fly species composition was performed using Shannon and CDC light traps. A total of 486 sand flies representing six Lutzomyia species were collected. L. evansi, L. panamensis and L. gomezi, known vectors of Leishmania spp. were the predominant sand fly species around dwellings. The finding of these species in relation to the appearance of the first cases of leishmaniasis in the city mentioned is discussed.     

Hidden Sylvatic Foci of the Main Vector of Chagas Disease Triatoma infestans: Threats to the Vector Elimination Campaign?

Background

Establishing the sources of reinfestation after residual insecticide spraying is crucial for vector elimination programs. Triatoma infestans, traditionally considered to be limited to domestic or peridomestic (abbreviated as D/PD) habitats throughout most of its range, is the target of an elimination program that has achieved limited success in the Gran Chaco region in South America.

Methodology/Principal Findings

During a two-year period we conducted semi-annual searches for triatomine bugs in every D/PD site and surrounding sylvatic habitats after full-coverage spraying of pyrethroid insecticides of all houses in a well-defined rural area in northwestern Argentina. We found six low-density sylvatic foci with 24 T. infestans in fallen or standing trees located 110–2,300 m from the nearest house or infested D/PD site detected after insecticide spraying, when house infestations were rare. Analysis of two mitochondrial gene fragments of 20 sylvatic specimens confirmed their species identity as T. infestans and showed that their composite haplotypes were the same as or closely related to D/PD haplotypes. Population studies with 10 polymorphic microsatellite loci and wing geometric morphometry consistently indicated the occurrence of unrestricted gene flow between local D/PD and sylvatic populations. Mitochondrial DNA and microsatellite sibship analyses in the most abundant sylvatic colony revealed descendents from five different females. Spatial analysis showed a significant association between two sylvatic foci and the nearest D/PD bug population found before insecticide spraying.

Conclusions

Our study shows that, despite of its high degree of domesticity, T. infestans has sylvatic colonies with normal chromatic characters (not melanic morphs) highly connected to D/PD conspecifics in the Argentinean Chaco. Sylvatic habitats may provide a transient or permanent refuge after control interventions, and function as sources for D/PD reinfestation. The occurrence of sylvatic foci of T. infestans in the Gran Chaco may pose additional threats to ongoing vector elimination efforts.     

Anthropogenic influence on the distribution, abundance and diversity of sandfly species (Diptera: Phlebotominae: Psychodidae), vectors of cutaneous leishmaniasis in Panama

In Panama, species of the genus Lutzomyia are vectors of American cutaneous leishmaniasis (ACL). There is no recent ecological information that may be used to develop tools for the control of this disease. Thus, the goal of this study was to determine the composition, distribution and diversity of Lutzomyia species that serve as vectors of ACL. Sandfly sampling was conducted in forests, fragmented forests and rural environments, in locations with records of ACL. Lutzomyia gomezi, Lutzomyia panamensis and Lutzomyia trapidoi were the most widely distributed and prevalent species. Analysis of each sampling point showed that the species abundance and diversity were greatest at points located in the fragmented forest landscape. However, when the samples were grouped according to the landscape characteristics of the locations, there was a greater diversity of species in the rural environment locations. The Kruskal Wallis analysis of species abundance found that Lu. gomezi and Lu. trapidoi were associated with fragmented environments, while Lu. panamensis, Lutzomyia olmeca bicolor and Lutzomyia ylephiletor were associated with forested environments. Therefore, we suggest that human activity influences the distribution, composition and diversity of the vector species responsible for leishmaniasis in Panama.     

Abundance of Lutzomyia ovallesi but not Lu. gomezi (Diptera: Psychodidae) correlated with cutaneous leishmaniasis incidence in north-central Venezuela

In north-central Venezuela Lutzomyia gomezi and Lu. ovallesi are the main endophilic/anthropophilic species of phlebotomine sandflies implicated as vectors of cutaneous leishmaniasis (CL). Lutzomyia ovallesi has been found infected with Leishmania braziliensis (1.2%) and less often with Le. mexicana (0.07%), while Le. braziliensis infections have also been found in Lu. gomezi (0.47%). We investigated population densities of these sandflies using two sampling methods with four series of collections between January 1991 and March 1995 at El Ingenio, Miranda State. All-night outdoor collections from a Shannon trap were correlated with indoor collections from CDC light–traps by linear regression, which proved to be statistically significant for both species. Estimated numbers of female sandflies per house per night were found to be proportional to monthly precipitation (i.e. rainfall), with a lag time of seven months for Lu. ovallesi and of six months for Lu. gomezi . Predominance of Lu. ovallesi over Lu. gomezi ( c. 10 :1 ) was observed throughout the year, with the number of infected females estimated as 0.043 ± 0.047 Lu. ovallesi and 0.0085 ± 0.0124 Lu. gomezi per CDC trap per house per night (ratio ∼ 5:1). The mean rate of new CL cases per house per year and sandfly abundance were correlated by linear regression, showing a statistically significant relationship for Lu. ovallesi but not for Lu. gomezi . The negative intercept indicated that, on average, the CDC trap density exceeds 800 Lu. ovallesi females/house/year before new CL cases occur at El Ingenio.     

Indicators of infestation, colonization and infection of Triatoma dimidiata (Latreille) (Hemiptera: Reduviidae) in Campeche, México

Triatoma dimidiata (Latreille) is considered to be one of the primary vectors of Chagas disease in Southern Mexico and Central America. The objective of the present study was to obtain ecological information on T. dimidiata in two rural communities of Campeche, Mexico, where the vector is poorly studied. Our work consisted of monthly samplings carried out during one-year time at three levels: sylvatic, peridomestic and intradomestic, in order to estimate the population abundance of this species and its rate of infection with Trypanosoma cruzi. Triatoma dimidiata was the unique vector of this disease collected in San Juan Bautista Sakcabchen (SJBS) and Crucero San Luis (CSL). The total of 145 individuals were captured in SJBS; from these, 26.9%, 20% and 53.1% were collected in the sylvatic, peridomestic and intradomestic area, respectively. In CSL captures yielded 108 individuals: 40.7% in the sylvatic area, 20.4% peridomestic and 38.9% intradomestic. We found no correlation between climatic variables and population abundance of T. dimidiata. Dataset obtained suggests that individuals from the sylvatic area exhibit a high rate of natural infection by T. cruzi, with monthly percentages up to 61.5% for SJBS and 50% for CSL. At the peri and intradomestic level, the reservoirs apparently play an important role in the transmission, as the seroprevalence in dogs was 61.5% y 65.4%, for SJBS y CSL, respectively. Based on these findings, it was concluded that inhabitants of both communities are at a high risk of Chagas disease infection.     

Sylvatic and peridomestic populations of Triatoma pseudomaculata are not significantly structured by habitat,as revealed by two genetic markers

Chagas disease remains a public health concern in Brazil and other Latin American countries, mainly due to the potential domiciliation of native triatomine species. We analyzed the genetic variability of Triatoma pseudomaculata in sylvatic and peridomestic ecotopes throughout three localities in the northeastern state of Bahia, Brazil. We studied polymorphisms generated by random amplified polymorphic DNA (RAPD) and isoenzyme electrophoresis analyses. Based on RAPD analysis, each specimen was assigned to one of three genetic clusters. Although all sylvatic specimens from one locality were grouped into the same cluster, sylvatic and peridomestic specimens from the other two localities were broadly distributed between the remaining two clusters, suggesting that geographic population structuring was not occurring. Furthermore, isoenzyme analysis suggested that distinct populations were in Hardy‐Weinberg equilibrium. Low statistical values for Wright's Fst index also supported the absence of population structuring and suggested the occurrence of panmixia. We conclude that genetic flow occurs between sylvatic and peridomestic T. pseudomaculata populations, probably as a consequence of passive and active dispersion of the insects, associated with deforestation and anthropic transformations.     

Morphometric study of the genus Panstrongylus Berg, 1879 (Hemiptera, Reduviidae, Triatominae)

The subfamily Triatominae is actually represented by 137 species distributed among 6 tribes and 19 genera. Within this subfamily, the genus Panstrongylus, Berg 1879, is composed by 13 species widespread in sylvatic, peridomestic, and domestic habitats of Neotropical regions. These species are vectors of Chagas disease and consequently are found associated with its main hosts, such as birds and mammals. Interest in species of this genus has been increasing in the last few years. Reports of several authors indicate these Triatominae to invade and colonize houses, increasing their epidemiological significance. Morphometry was used in this study to investigate correlations among possible closely related species. We measured 224 specimens among 13 species through a set of metric variables of the head. The results indicated that the genus Panstrongylus seems to be homogeneous since 10 of the 14 species were shown to be closely related.     

Ecology of sandflies (Diptera:Psychodidae) in a restricted focus of cutaneous leishmaniasis in northern Venezuela. IV. Sandfly monthly fluctuation and leishmaniasis incidence relationship

An attempt has been made to correlate the monthly incidence of human leishmaniasis with the temporal distribution of sandfly species at San Esteban, Northern Venezuela. Upon statistical analysis, the seasonal fluctuation of L. ovallesi population correlated strongly with the human disease, while the dynamics of L. panamensis, generally believed to be the vector in the Central area of the country, showed only a very weak correlation. These findings support the hypothesis that L. panamensis might not be the main or unique species responsible for the transmission in this area and that L. ovallesi and additionally L. olmeca bicolor might be involved in the epidemiology of the disease.     

Sex, subdivision, and domestic dispersal of Trypanosoma cruzi lineage I in southern Ecuador

Background

Molecular epidemiology at the community level has an important guiding role in zoonotic disease control programmes where genetic markers are suitably variable to unravel the dynamics of local transmission. We evaluated the molecular diversity of Trypanosoma cruzi, the etiological agent of Chagas disease, in southern Ecuador (Loja Province). This kinetoplastid parasite has traditionally been a paradigm for clonal population structure in pathogenic organisms. However, the presence of naturally occurring hybrids, mitochondrial introgression, and evidence of genetic exchange in the laboratory question this dogma.

Methodology/Principal Findings

Eighty-one parasite isolates from domiciliary, peridomiciliary, and sylvatic triatomines and mammals were genotyped across 10 variable microsatellite loci. Two discrete parasite populations were defined: one predominantly composed of isolates from domestic and peridomestic foci, and another predominantly composed of isolates from sylvatic foci. Spatial genetic variation was absent from the former, suggesting rapid parasite dispersal across our study area. Furthermore, linkage equilibrium between loci, Hardy-Weinberg allele frequencies at individual loci, and a lack of repeated genotypes are indicative of frequent genetic exchange among individuals in the domestic/peridomestic population.

Conclusions/Significance

These data represent novel population-level evidence of an extant capacity for sex among natural cycles of T. cruzi transmission. As such they have dramatic implications for our understanding of the fundamental genetics of this parasite. Our data also elucidate local disease transmission, whereby passive anthropogenic domestic mammal and triatomine dispersal across our study area is likely to account for the rapid domestic/peridomestic spread of the parasite. Finally we discuss how this, and the observed subdivision between sympatric sylvatic and domestic/peridomestic foci, can inform efforts at Chagas disease control in Ecuador.     

Ecotope effect in Triatoma brasiliensis (Hemiptera: Reduviidae) suggests phenotypic plasticity rather than adaptation

Triatoma brasiliensis (Hemiptera: Reduviidae) is an important vector of Chagas' disease in both sylvatic and peridomestic ecotopes. Discriminating between these populations of Triatominae has been proposed as a means of investigating re‐infestation rates of human dwellings. Geometric morphometrics have been widely applied in the study of Triatominae polymorphisms at species and population levels. This study characterizes morphometric differences between sylvatic and peridomestic populations, as well as between sexes in T. brasiliensis specimens from Jaguaruana, Ceará, in northeastern Brazil. No differences in either the shape or size of the cephalic capsule were apparent between sexes or ecotopes. However, the wings showed differentiation in shape and size. Sexual dimorphism was detected, with females presenting significantly higher values and conformations. Size differentiation was also evident, with sylvatic specimens being generally larger than peridomestic examples. These results indicate that differences in the wings of T. brasiliensis may be related to the existence of phenotypic plasticity, and variations in size and shape may be associated with different ecotopes, possibly as a result of conditions in each micro‐habitat, such as temperature, relative humidity, food supply and density.     

Limited catching bias in a wild population of birds with near‐complete census information

Animal research often relies on catching wild animals; however, individuals may have different trappability, and this can generate bias. We studied bias in mist netting, the main method for catching wild birds. The unusually high resighting rate in our study population—house sparrows (Passer domesticus) on Lundy Island (England)—allowed us to obtain accurate estimates of the population size. This unique situation enabled us to test for catching bias in mist netting using deviations from the expected Poisson distribution. There was no evidence that a fraction of the birds in the population consistently remained uncaught. However, we detected a different bias: More birds than expected were captured only once within a year. This bias probably resulted from a mixture of fieldworkers sometimes ignoring rapid recaptures and birds becoming net shy after their first capture. We had sufficient statistical power with the available data to detect a substantial uncaught fraction. Therefore, our data are probably unbiased toward catching specific individuals from our population. Our analyses demonstrate that intensively monitored natural insular populations, in which population size can be estimated precisely, provide the potential to address important unanswered questions without concerns about a fraction of the population remaining uncaught. Our approach can help researchers to test for catching bias in closely monitored wild populations for which reliable estimates of population size and dispersal are available.     

Extracellular amastigote-like forms of Leishmania panamensis and L. braziliensis. II. Stage- and species-specific monoclonal antibodies

Immunochemical evidence, employing monoclonal antibodies, shows that the forms of L. braziliensis complex axenically grown at elevated temperature are amastigote-like. The monoclonal antibodies were raised against membrane proteins of amastigote-like forms, strains of both L. panamensis (WR442) and L. braziliensis (M5052), which were grown axenically. The specificities of these antibodies were examined by indirect radioimmune binding assay, indirect immunofluorescent assay and Western blot analyses. Two distinct groups of monoclonal antibodies were obtained and their specificities were consistent with the 3 methods used. Four antibodies are specific for the species L. panamensis and react with both developmental stages. Six antibodies specifically recognize amastigote-like forms grown at elevated temperature and intracellular amastigotes of both L. panamensis (WR442) and L. braziliensis (M5052). These monoclonal antibodies do not bind to promastigotes of these species, nor to promastigotes of any other species of Leishmania. Therefore these antibodies are specific for amastigotes of L. panamensis (WR442) and L. braziliensis (M5052), and suggest that immunochemically both amastigote forms (culture and macrophage) are developmentally very close, if not identical. The molecules associated with the amastigote-specific antigenic determinants consist of a Mr 12-kD component and a heterogeneous component (Mr from 50 kD to greater than 200 kD); these molecules appear to be identical for both amastigote-like forms and amastigotes isolated from macrophages.     

Extracellular Amastigote-Like Forms of Leishmania panamensis and L. braziliensis. II. Stage- and Species-Specific Monoclonal Antibodies

Immunochemical evidence, employing monoclonal antibodies, shows that the forms of L. braziliensis complex axenically grown at elevated temperature are amastigote-like. The monoclonal antibodies were raised against membrane proteins of amastigote-like forms, strains of both L. panamensis (WR442) and L. braziliensis (M5052), which were grown axenically. The specificities of these antibodies were examined by indirect radioimmune binding assay, indirect immunofluorescent assay and Western blot analyses. Two distinct groups of monoclonal antibodies were obtained and their specificities were consistent with the 3 methods used. Four antibodies are specific for the species L. panamensis and react with both developmental stages. Six antibodies specifically recognize amastigote-like forms grown at elevated temperature and intracellular amastigotes of both L. panamensis (WR442) and L. braziliensis (M5052). These monoclonal antibodies do not bind to promastigotes of these species, nor to promastigotes of any other species of Leishmania . Therefore these antibodies are specific for amastigotes of L. panamensis (WR442) and L. braziliensis (M5052), and suggest that immunochemically both amastigote forms (culture and macrophage) are developmentally very close, if not identical. The molecules associated with the amastigote-specific antigenic determinants consist of a Mr 12-kD component and a heterogeneous component (Mr from 50 kD to >200 kD); these molecules appear to be identical for both amastigote-like forms and amastigotes isolated from macrophages.     

Multiplex-PCR for detection of natural Leishmania infection in Lutzomyia spp. captured in an endemic region for cutaneous leishmaniasis in state of Sucre, Venezuela

We studied the natural infection of Lutzomyia (Lutzomyia) sp. with Leishmania in endemic foci of cutaneous leishmaniasis in the Paria peninsula, state of Sucre, Venezuela. Sand flies were collected between March 2001 and June 2003, using Shannon light-traps and human bait. Of the 1291 insects captured, only two species of phlebotomines were identified: L. ovallesi (82.75%) and L. gomezi (17.42%). A sample of the collected sand flies (51 pools of 2-12 individuals) were analyzed by using a multiplex-PCR assay for simultaneous detection of New Word Leishmaniaand Viannia subgenera. The results showed a total of 8 pools (15.68%) infected; of these, 7 were L. ovallesi naturally infected with L. braziliensis (2 pools) and L. mexicana (5 pools) and 1 pool of L. gomezi infected by L. braziliensis.