Feeding behaviour of Belminus ferroae (Hemiptera: Reduviidae), a predaceous Triatominae colonizing rural houses in Norte de Santander, Colombia.

Belminus ferroae (Hemiptera: Reduviidae) was collected from houses in the localities of San Alberto and Santa Catalina in the municipality of Toledo, Department of Norte de Santander, Colombia. Precipitin tests revealed that the main hosts of B. ferroae are insects belonging to the order Blattodea, whose haemolymph serves as food (89.43%). A low rate of haematophagy was indicated on rodents, dogs and humans (7.32%) and no natural infection with Trypanosoma cruzi was found by either microscopic examination or polymerase chain reaction. The finding of B. ferroae inside houses, feeding primarily on Blattodea with only a minor tendency to feed on vertebrate blood, is indicative of both the evolutionary path from predator to blood-sucker taken by Triatominae and their tendency to adapt from silvatic to domestic habitats.

Epidemiology of American tegumentary leishmaniasis in domestic dogs in an endemic zone of western Venezuela.

Domestic dogs are not only reservoir hosts of the American zoonotic visceral leishmaniasis (ZVL) but of the American zoonotic tegumentary leishmaniasis (ATL) as well, for different reasons. However it is still controversial to state that dogs are incriminated as ATL reservoir hosts as there is evidence that humans and dogs are likely to be exposed in the same way to sandfly vector. In Venezuela this issue has not been completely addressed, for this reason we selected a location inside Trujillo city to study eco-epidemiological conditions as well as to survey a significant sample of dogs by Montenegro Skin Test (MST). Antigen was prepared according to standard procedure using Leishmania (V) braziliensis promastigotes (80 microg/ml); response was read 48 hours post-inoculation with an induration size > 5 mm being considered as positive. The study place is an endemic mountainous semi-urban area located at 850-950 masl with an average rainfall of 150 mm/year. We evaluated 61 dogs in 46 houses with 168 human beings. Among the human population 27 cases of ATL were reported (16.1%). With the MST we found 19 positive-reaction dogs (31%) (mean MST size of 9.58 mm, 95% CI: 8.41-10.75) in 13 houses (28%). Multivariate analysis did not reveal significant association between domestic MST positive-dog ownership and human ATL cases (RR = 1.48, p = 0.28). Although some studies have indicated that dog ownership and dog infection rates are associated with an increased risk of human disease in different evaluated places, this question has not been completely answered in Venezuelan studied zones, further research is necessary.

Diversity and species composition of sand flies (Diptera: Psychodidae) in a Venezuelan urban focus of cutaneous leishmaniasis.

The present study examined the spatial and temporal abundance and diversity of phlebotomine sand flies in an area of Venezuela that is an ancient focus of leishmaniasis. The study was conducted in 6 stations in urban localities in Trujillo City, located in northwestern Venezuela (9 degrees 22' 24" N, 70 degrees 26' 08" W), which is located in a mountain range in the Andean ecoregion (altitude = 600-1,010 m). During 1995-99, entomological surveys were conducted after and before the rainy season. Shannon light traps were operated from 1800 to 2000 h in peridomestic site trap locations. Twelve species were captured, and Lutzomyia youngi, L. ovallesi, L. scorzai, L. gomezi, L. lichyi, and L. shannoni occurred at all localities in each year. The abundance of these species showed low variation over time but high variation between localities. The Sørensen similarity index, used to compare diversity between years within each locality, ranged from 0.60 at Carmona to 0.84 at La Hacienda. Sand fly communities exhibited annual variation in species richness and diversity. Variations were affected more by changes in species abundance than by changes in species composition. Lutzomyia ovallesi, L. lichyi, and L. scorzai had the highest coefficient of variation between years (63, 38, and 23%, respectively).

Acute Chagas' disease in western Venezuela: a clinical, seroparasitologic, and epidemiologic study.

A clinical, parasitologic, and serologic study carried out between 1988 and 1996 on 59 acute-phase patients in areas of western Venezuela where Chagas' disease is endemic showed 19 symptomatic patterns or groups of symptoms appearing in combination with different frequencies. The symptomatic pattern with the highest frequency was that showing simultaneously fever, myalgia, headache, and Romaña's sign, which was detected in 20% of the acute-phase patients. Asymptomatic individuals and patients with fever as the only sign of the disease made up 15% and 11.9% of the total acute cases, respectively. Statistical correlation analysis revealed that xenodiagnosis and hemoculture were the most reliable and concordant of the five parasitologic methods used; these two methods also showed the highest proportions in detecting any clinical symptomatic pattern in acute-phase patients. A similar high reliability and concordance was obtained with a direct agglutination test, an indirect immunofluorescent antibody test, and an ELISA as serologic tests, which also showed a higher proportion of positive detection of clinical patterns than parasitologic methods (P < 0.001). It is recommended that individuals coming from endemic areas showing mild and/or severe clinical manifestations should be suspected of being in contact or having been in contact with Trypanosoma cruzi, be referred for parasitologic and serologic evaluations to confirm the presumptive clinical diagnosis of acute Chagas' disease, and start specific treatment. The epidemiologic implications of the present findings are discussed and the use of similar methodology to evaluate other areas where Chagas' disease is endemic is suggested.

[Presence of metacyclical promastigotes of Leishmania pifanoi in the hypopharynx of Lutzomyia youngi and their sugar consumption]. / Presencia de nectomonadas metacíclicas de L. pifanoi en la hipofaringe de Lutzomyia youngi y la ingestión de azúcares.

The presence of metacyclical promastigotes of Leishmania pifanoi in the hypopharyngeal duct of Lutzomyia youngi is reported. The insects were experimentally infected by engorgement on the tarsal lesions of hamsters. The metacyclics, whose morphology is illustrated, appeared in the hypopharynx 5 to 9 days after engorgement; they were more frequently found in the insects fed on unrefined commercial sugar. They role of amino derivates of glucose and galactose, as well as of amino acids in the development and migration of metacyclics, is investigated.

[Excretion of promastigotes of Leishmania pifanoi by experimentally infected Lutzomyia youngi]. / Excreción de promastigotos de Leishmania pifanoi por Lutzomyia youngi experimentalmente infectada.

The increase in the promastigotes population of Leishmania pifanoi in Lutzomyia youngi experimentally infected and kept on 50% sacarose under constant conditions of temperature and humidity is described. Two stages in the differentiation and growth of the parasites are recognised between two and twenty-four hours after meals. The pleomorphic differentiation of the amastigotes in short promastigotes which multiply by binary division for 60 hours, when the rupture of the peritrophic membrane occurs, takes place within 48 hours. The second stage occurs between 72 and 96 hours when some of the parasites migrate to the esophagic valve and the rest of the free parasites are excreted in fecal drops as large, active promastigotes. The first drops excreted react positively to glucose or contain crystals of urate. The excess promastigotes of the second stage of development are eliminated in the last excretions and react positively to the Hemoscreen and Biuret tests for total proteins as also for glucose and account for 82% of the of drops excreted. The excretion of parasites by Lu. youngi is a normal stage in the growth of L. pifanoi into a vector.

[Leishmania braziliensis: isolation from lesions by inoculation of hamsters with and without addition of salivary gland lysate from Lutzomyia youngi]. / Leishmania braziliensis: aislamiento de lesiones por inoculación de hámsteres con o sin adición de lisado de glándulas salivares de Lutzomyia youngi.

Homogenized biopsy tissue from the cutaneous leishmaniasis lesions of 50 patients from Trujillo, Venezuela, were inoculated subcutaneously into the tarsi of male hamsters. Homogenized tissue either alone or mixed with salivary gland lysates of Lutzomyia youngi were used for inoculation. Homogenized tissue alone yielded 58.5% of infections with a mean of twelve weeks for prepatency, while those mixed with sandfly lysate resulted in 92% of infections with a mean prepatency of three weeks.

Physiological age in Lutzomyia youngi (Diptera: Psychodidae) populations from an endemic area for cutaneous leishmaniasis, Venezuela.

Batches of sylvatic females of Lutzomyia youngi (Phlebotominae) captured in a Shannon trap on twelve occasions over one year in a locality where subcutaneous leishmaniasis is endemic, near the city of Trujillo, Venezuela, were used to study: 1) the percentages of parous females according to previously established criteria and 2) the average number of eggs laid spontaneously by isolated females during 7 days after feeding on hamsters. The data on the batches of females captured on nights previous to the rainy period (prepluvial) were compared with those on females captured after the rains (postpluvial). Significant differences were detected by variation analysis for two variables and different number of N, as also were consistent groupings by Duncan's Test for pre- and postpluvial lots of females. The females captured on nights prior to the rainy periods (January-March and August-September) presented higher rates of nulliparity (86-72%) and contained or laid a greater number of eggs (71-67) than those captured after the rains (March-June and November-December) which presented lower rates of nulliparity (60-24%) and a smaller number of eggs (50-30). The rainfall peaks occurred in April and September-October, respectively. It is considered that these differences can be used by epidemiological studies as a means of estimating the physiological age of female populations of L. youngy.

Presence of Leishmania braziliensis in blood samples from cured patients or at different stages of immunotherapy.

By using a PCR assay specific for Leishmania braziliensis in blood samples, we detected this parasite in patients cured by immunotherapy or at different stages of treatment. We also found the parasite in subjects who had never suffered leishmaniasis but who had lived in endemic areas and migrated to nonendemic ones many years ago. These results suggest that L. braziliensis infections are difficult to eradicate and that a clinical cure but rarely a complete elimination of the parasite is generally accomplished.

Identification and detection of Trypanosoma cruzi by using a DNA amplification fingerprint obtained from the ribosomal intergenic spacer.

We designed a PCR assay targeted on repeated elements of the ribosomal intergenic spacer which produces highly polymorphic DNA band patterns for different strains of Trypanosoma cruzi. By labeling the PCR products with digoxigenin and by chemiluminescence detection, we improved the assay sensitivity by three orders of magnitude to get T. cruzi strain fingerprints in feces of the trypanosome-infected triatomine bug vector. We also developed a capture assay for the digoxigenin-labeled PCR products that allowed us to detect T. cruzi in triatomine bug vector feces and in human serum samples with a solid support.

Identification of new world Leishmania using ribosomal gene spacer probes.

DNA probes from the nontranscribed ribosomal spacer (NTS), of Leishmania garnhami and Leishmania braziliensis were constructed and tested for sensitivity and specificity against different Leishmania isolates. The L. garnhami probes were species-specific under hybridization conditions of high stringency, but displayed specificity for the mexicana complex under conditions of intermediate stringency. The L. braziliensis probes showed 'complex' specificity. RFLP for the nontranscribed spacer within the braziliensis complex revealed very homogeneous patterns even for organisms currently accepted as different species. A PCR assay for the detection of Leishmania from the braziliensis complex is presented.

Populations of larvae of anopheles spp. in natural breeding sites in western Venezuela, an area of refractory malaria.

Studies have been undertaken into on the diversity and relative abundance of larvae of Anopheles (Nyssorhynchus) spp. in 22 permanent or temporary pools in an area of 70 km2 in the eastern piedmont of the Venezuela Andes, between the mountains and the plains, an area in which malaria is refractory and A. nuñeztovari is present. Twelve species were identified, the most frequent, abundant and sympatric being A. triannulatus, A. albitarsis, A. nuñeztovari, A. oswaldoi and A. strodei. The samples from the permanent pools showed greater diversity of species and greater numbers of larvae than the samples from the temporary pools. The existence of the same larval associations in pools of other localities in the eastern piedmont of the Venezuelan Andes suggests the possibility of the making an ecological map of the breeding sites of A. nuñeztovari and for these anophelines in a region extending for 430 km.

Leishmania braziliensis spp. in the nasal mucosa of guinea pigs inoculated in the tarsi.

Two lots of 20 young male guinea pigs were inoculated subcutaneously in the tarsi with 10(4) amastigotes of Leishmania braziliensis braziliensis or L. b. guyanensis to study the susceptibility of this Neotropical hystricomorph rodent the autochthonous parasites. Almost 50% of the animals showed lesions in the inoculation site and had parasitizations that were infective to hamsters, as shown by inoculating homogenates of the dermal lesion, of the spleen, of the liver, and of the nasal mucosa into hamsters at 20, 40, 60, and 120 days after inoculation of the guinea pig. Smears of the above organs showed the presence of amastigotes. Parasites inoculated into the tarsi were detected early in the skin, spleen, and liver of the guinea pig host. Blood cultures made by cardiopuncture on sacrifice of the guinea pigs were uniformly negative. The nasal mucosa of nearly all animals positive in the skin or viscera was invaded early by the parasites, although with greater frequency between 60 and 120 days post-inoculation. The use of this model for the study of mucocutaneous parasitism by L. braziliensis is discussed, together with the phenomena of parasitism at a distance from the inoculation site, the temperature of the body regions affected, and the possible genetic influence on susceptibility of the guinea pig to L. braziliensis.

[Metacyclics of Leishmania mexicana in the hypopharynx of Lutzomyia youngi experimentally infected]. / Metaciclicos de Leishmania mexicana en la hipofaringe de Lutzomyia youngi experimentalmente infectada.

Although it has been postulated that the natural transmission of leishmaniasis occurs by regurgitation of the parasites from contaminated proboscis of phlebotomine vector recent experimental results seems to oppose this thesis. Wild-caught and laboratory-reared females of Lutzomyia youngi, vector of Leishmania mexicana in the Venezuelan Andes, were infected on tarsal lesion of hamsters, inoculated 6 weeks previously s.c. with 2 x 10(4) amastigotes of L. mexicana s.l., isolated from Venezuelan case of diffuse leishmaniasis. The insects were kept at 23 degrees C and 80% R.H., on a 50% (v:v) solution of "unrefined" cane sugar. The parasites developed through 3 stages: i) differentiation of the amastigotes and exponential growth in the number of promastigotes, ending between 60 and 108 hrs, with a massive fecal elimination of large promastigotes; ii) a stationary phase in growth with flagellar adhesion to the cuticle of the stomodeum and synthesis of a gel that formed a plug between 60 and 120 hrs; iii) differentiation of metacyclics, which invade the hypopharyngeal duct in 7% of the insects, from the 5th day post-infection, and most frequently between the 6th and 9th day. The metacyclics measured 4.2/microns in body length, 1.07/microns in maximum with and the flagellum was 8.8/microns long. The parasites swam freely in the saliva of the hypopharyngeal duct (lumen 6/microns in diameter), from the apex to union with the salivary duct, without invading the latter. Similar results has been obtained in the same vector experimentally infected with other two strains of L. mexicana isolated respectively from wild Lu. flaviscutellata and from another case of diffuse leishmaniasis.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunogenetics of human American cutaneous leishmaniasis. Study of HLA haplotypes in 24 families from Venezuela.

Twenty-four families with one or multiple cases of localized cutaneous leishmaniasis (LCL) from an endemic region with the highest incidence of LCL in Venezuela were typed from HLA-ABC, DR, DQ antigens and complement factors. The parental HLA haplotypes segregated at random among healthy and affected siblings but in backcross families significantly higher frequencies of HLA-A28 (p = 0.0018), -Bw22 (p = 0.0122), or -DQw8 (p = 0.0364) were present in affected compared to healthy siblings. HLA-B15 showed a higher frequency (p = 0.0062) among the latter group. Haplotypes Bw22CF31 (p = 0.0076) and Bw22DRw11DQw7 (p = 0.0163) were also significantly more frequent in affected compared to healthy siblings and A2Cw- (p = 0.0445) among the latter. No HLA genetic linkage with a putative LCL susceptibility gene(s) could be demonstrated in this study. A case/control comparison of 26 unrelated LCL patients (one proband from each family) and healthy individuals of the same ethnic origin confirmed the association of HLA-Bw22 (pc = 0.048) and -DQw3 (pc = 0.036) with LCL. The relative risk reached 12.5 for Bw22 and 4.25 for DQw3 with ethiologic factors of 0.17 and 0.64, respectively. HLA-DQw3 apparently makes the major contribution as a genetic risk factor for LCL at the population level.

[Xenodiagnosis using Lutzomyia youngi in Venezuelan cases of cutaneous leishmaniasis due to Leishmania braziliensis]. / Xenodiagnostico con Lutzomyia youngi en casos venezolanos de leishmaniasis cutanea por Leishmania braziliensis.

Eight patients infected with Leishmania braziliensis were used for xenodiagnosis with Lutzomyia youngi, before and after specific antileishmanial treatment with "glucantime" and "gabbromycin". All of them infected sandflies fed on the borders of the skin lesions before the treatment, suggesting that infected persons might act as reservoirs of infection for an indoor-biting sandfly species. The negative results obtained by xenodiagnosis carried out after specific treatment of the same individuals indicated cure of the patients, and a reduction of risk for further intradomiciliary transmission.