Attempts to Improve Xenodiagnosis: Comparative Test of Sensibility Using Rhodnius neglectus, Panstrongylus megistus, Triatoma vitticeps and Triatoma infestans in Endemic Areas of Brazil

From June 1984 to July 1992, 392 xenodiagnostic tests were applied on 264 patients with chronic Chagas disease from Brazilian endemic areas of Virgem da Lapa and Coronel Murta, situated in the Jequitinhonha Valley, in the State of Minas Gerais. The susceptibilities of Rhodnius neglectus, Panstrongylus megistus, Triatoma vitticeps and Triatoma infestans were compared. Most of the time 20 nymphs (fourth instar) of each species were applied to 161 women and 103 men aged between 5 and 83 years of age. The tests were prepared to compare the susceptibilities of two species at a time, using the same patients for each test. Results showed a xenopositiveness of 26.28% (103 tests) being 27.98% in women (68 positive in 243 applied tests) and 23.49% in men (35 positive in 149 applied tests). The relative frequency of xenopositiveness displayed a great superiority of P. megistus and T. vitticeps. In tests from type I, for example, P. megistus was the unique responsible for 10.73% of positive xenodiagnosis vs. only 0.98% in T. infestans. Other parameters analized in this work confirm this superiority, and corroborate that T. infestans can be replaced by P. megistus and /or T. vitticeps in order to upgrade the efficacy of xenodiagnosis

Variations of the External Male Genitalia in Three Populations of Triatoma infestans Klug, 1834

Triatoma infestans is the triatomine that presents the greatest dispersion area in South America. However, it is not known whether the original characteristics of this insect remained in its long dispersion process. The purpose of this work was to study comparatively the external male genitalia of insects from different populations of T. infestans, two from Brazil (Minas Gerais and Bahia) and one from Bolivia (Cochabamba Valley), and to investigate the correlation between the morphological and behavioral variations. Differences were observed in one of the structures of the external genitalia (endosoma process) that could be used to characterize the insects from the three populations studied.

Comparative developmental and susceptibility to insecticide of Bolivian and Brazilian populations of Triatoma infestans

The triatomine bug Triatoma infestans probably originated in Bolivia and dispersed passively over wide areas of South America, where it is the principal vector of Trypanosoma cruzi. In the region of its probable origin this species shows colonization in two different ecotopes, so that it may be encountered in sylvatic as well as in artificial habitats. The sylvatic colonization pattern is not observed in the rest of its range, where T. infestans is exclusive to man-made habitats. The objective of this study was to compare several aspects of two T. infestans populations, one from Minas Gerais (Brazil) and the other from the Cochabamba Valley (Bolivia), with a view to elucidate the factors associated with the different colonization patterns observed for this species. The differences between the developmental cycle, weight, capacity to ingest blood and mortality rate of first instar nymphs should indicate more fragility of Brazilian population that may be related to its elimination possibility.

Genetic variability among populations of Lutzomyia (Psathyromyia) shannoni (Dyar 1929) (Diptera: Psychodidae: Phlebotominae) in Colombia

Polyacrylamide gel electrophoresis was used to elucidate genetic variation at 13 isozyme loci among forest populations of Lutzomyia shannoni from three widely separated locations in Colombia: Palambí (Nariño Department), Cimitarra (Santander Department) and Chinácota (Norte de Santander Department). These samples were compared with a laboratory colony originating from the Magdalena Valley in Central Colombia. The mean heterozygosity ranged from 16 to 22%, with 2.1 to 2.6 alleles detected per locus. Nei's genetic distances among populations were low, ranging from 0.011 to 0.049. The estimated number of migrants (Nm=3.8) based on Wright's F-Statistic, F ST, indicated low levels of gene flow among Lu. shannoni forest populations. This low level of migration indicates that the spread of stomatitis virus occurs via infected host, not by infected insect. In the colony sample of 79 individuals, the Gpi locus was homozygotic (0.62/0.62) in all females and heterozygotic (0.62/0.72) in all males. Although this phenomenon is probably a consequence of colonization, it indicates that Gpi is linked to a sex determining locus.

A soil emergence trap for collections of phlebotomine sand flies

The identification of breeding sites of sand flies is of great epidemiological interest. A soil emergence trap for investigating potential sand fly breeding sites is described. The trap was tested in two rural areas in the Mogi Guaçu River Valley where the American cutaneous leishmaniasis is an endemic disease. Seventy-three sand fly individuals of three species, Lutzomyia intermedia s. l., L. whitmani and L. pessoai, were collected on the forest floor and peridomicile.

Distribution of sand flies (Diptera: Psychodidae) at different altitudes in an endemic region of American cutaneous leishmaniasis in the State of Espírito Santo, Brazil

The involvement of different sand fly species in the transmission of American cutaneous leishmaniasis (ACL) at different altitudes was evaluated in the municipality of Afonso Cláudio in the State of Espírito Santo, Brazil, from November 1995 to February 1997. CDC light traps and Shannon traps baited with human volunteers were hung simultaneously at three altitudes of a river valley: (a) 650-750 m in an area of active ACL transmission; (b) 750-850 m in a transitional area; and (c) 850-950 m in an area where no ACL transmission occurred. A total of 13,363 specimens belonging to 28 species was collected. The five most abundant man-biting species were Lutzomyia intermedia, which constituted 24.3% of the total, Lu. migonei (22.3%), Lu. whitmani (15.4%), Lu. fischeri (14.9%) and Lu. monticola (5.8%). Analysis of the distribution of these species at the three altitudes provided evidence that Lu. fischeri and Lu. monticola were not involved in ACL transmission, whereas Lu. migonei and Lu. whitmani might act as secondary vectors of Leishmania (Viannia) braziliensis and Lu. intermedia was probably the principal vector of the parasite. These results reinforce those published in the existing literature, which indicate that Lu. intermedia is the main vector of Le. (V.) braziliensis in Southeast Brazil, while Lu. migonei and Lu. whitmani are of secondary importance.

Altered Response of Strain of Schistosoma mansoni to Oxamniquine and Praziquantel

The susceptibility of a fourth generation Ouh strain (Paranapanema Valley, São Paulo, Brazil) of Schistosoma mansoni to oxamniquine (OXA) and praziquantel (PZQ) was studied. Ten groups of 13 female albino mice each were infected with 70 cercariae per animal. These mice were medicated orally on the 50th day after infection. Five groups were given OXA doses of 0, 100, 200, 300 and 400 mg/kg (single doses) and the rest were treated with PZQ doses of 0, 100, 200, and 250 mg/kg/5 days. Each group was sub-divided: 8 animals underwent perfusion after 15 days treatment, 5 mice followed up for oviposition and their feces were tested every 15 days for miracidia hatching. The efficacy of the OXA doses of 100 and 200 mg/kg was 66% and 91.4%, respectively and for the 100 mg/kg PZQ dose it was 90.1%. The follow-up groups with 100 and 200 mg/kg of OXA and PZQ, 100 and 150 mg/kg, showed that they re-established the oviposition after a period of 60 to 75 days of treatment. The ED50 was 69.6mg/kg OXA and 39.4 mg/kg PZQ. The results show the tolerance of the Ouh strain to a dose of 100 mg with both drugs and they appoint the need for a dose review during the follow up of the oviposition and in monitoring phenomena in the field.

Inca expansion and parasitism in the Lluta Valley: preliminary data

Assessing the impact of cultural change on parasitism has been a central goal in archaeoparasitology. The influence of civilization and the development of empires on parasitism has not been evaluated. Presented here is a preliminary analysis of the change in human parasitism associated with the Inca conquest of the Lluta Valley in Northern Chile. Changes in parasite prevalence are described. It can be seen that the change in life imposed on the inhabitants of the Lluta Valley by the Incas caused an increase in parasitism.

In vivo binding of the Cry11Bb toxin of Bacillus thuringiensis subsp. medellin to the midgut of mosquito larvae (Diptera: Culicidae)

Bacillus thuringiensis subsp. medellin produces numerous proteins among which 94 kDa known as Cry11Bb, has mosquitocidal activity. The mode of action of the Cry11 proteins has been described as similar to those of the Cry1 toxins, nevertheless, the mechanism of action is still not clear. In this study we investigated the in vivo binding of the Cry11Bb toxin to the midgut of the insect species Anopheles albimanus, Aedes aegypti, and Culex quinquefasciatus by immunohistochemical analysis. Spodoptera frugiperda was included as negative control. The Cry11Bb protein was detected on the apical microvilli of the midgut epithelial cells, mostly on the posterior midgut and gastric caeca of the three mosquito species. Additionally, the toxin was detected in the Malpighian tubules of An. albimanus, Ae. aegypti, Cx. quinquefasciatus, and in the basal membrane of the epithelial cells of Ae. aegypti midgut. No toxin accumulation was observed in the peritrophic membrane of any of the mosquito species studied. These results confirm that the primary site of action of the Cry11 toxins is the apical membrane of the midgut epithelial cells of mosquito larvae.

First report of Lymnaea cousini Jousseaume, 1887 naturally infected with Fasciola hepatica (Linnaeus, 1758) (Trematoda: Digenea) in Machachi, Ecuador

We report the first finding of Lymnaea cousini naturally infected with Fasciola hepatica in Ecuador. A sample of 70 snails was collected in April 2005 from a wetland located in a valley at approximately 3000 m a.s.l., near the locality of Machachi, Pichincha Province. The prevalence of natural infection in L. cousini was 31.43%, which is the highest value ever recorded for naturally infected lymnaeid species.

Anatomical studies of sibling species within neotropical lymnaeids, snail intermediate hosts of fascioliasis

Several anatomical parameters of the reproductive system have been used to distinguish Lymnaea cubensis from L. viatrix, the snail hosts of fascioliasis in South America and the Caribbean area. Three samples have been collected in the type localities of L. cubensis (Cuba), L. viatrix var. A ventricosa (Argentina, Río Negro Lower Valley), and L. viatrix var. B elongata (Peru, Lima), respectively. Only one parameter, the relative lengths of the penis sheath and preputium, showed significant differences between L. viatrix var. ventricosa and the two other taxa. None of the studied parameters separated L. cubensis from L. viatrix var. elongata.

Biology of three species of the Meccus phyllosomus complex (Hemiptera: Reduviidae: Triatominae) fed on blood of hens and rabbits

Aspects related to hatching, life time, number of blood meals to molt, mortality, feeding time and postfeed defecation delay for each instar of Meccus phyllosomus, M. mazzottii, and M. bassolsae, life-cycle were evaluated and compared in two cohorts of each of those three species, fed on hens or rabbits. No significant (p > 0.05) differences were recorded among cohorts fed on hens respect to cohorts fed on rabbits in M. phyllosomus and M. mazzottii and the average time of hatching was 21.5 days for cohorts fed on hens and 22.5 for cohorts fed on rabbits. Average egg-to-adult development times were no significant (p > 0.05) different between both cohorts of M. phyllosomus and M. mazzotti, independent of the blood meal source. The average span in days for each instar fed on hens was not significantly different to the average span for each instar fed on rabbits, when comparisons were made by species. The number of blood meals at each nymphal instar varied from 1 to 6 in both cohorts of each species. The mortality rates were higher on older nymphs, in both cohorts of M. phyllosomus and M. bassolsae, whereas they were higher on first instar nymphs on M. mazzottii. Mean feeding time was no significant (p > 0.05) different in triatomines fed on hens or fed on rabbits, when each species were compared separately. A similar number of nymphs of each cohort, completed the cycle. Defecation delay was no significant (p > 0.05) different when cohorts fed on hens and fed on rabbits were compared by species. Most of the studied parameters showed no significant (p > 0.05) differences among those cohorts fed on hens and for fed on rabbits, which could mean a high degree of association of those species with birds as much as mammals, under wild conditions, increasing their capacity to colonize human dwellings.

Biomphalaria tenagophila potencial vector of Schistosoma mansoni in the Paraná River basin (Argentina and Paraguay)

Susceptibility and compatibility experiments were carried out with 700 Biomphalaria tenagophila from the Paraná River basin exposed to infection with Schistosoma mansoni. Individual infection was performed with 10 miracidia of SJ2 strain from the Paraiba valley (Brazil) originally infective to B. tenagophila. These snails were laboratory-breed progeny of B. tenagophila collected from six localities of Argentina and one from Paraguay. From Argentina: Rincón de Vences (7%) and Posadas (11%) became infected with S. mansoni and the calculation of Frandsen's index (TCP/100) shows that they were Class II poorly compatible. Those snails from Goya (22%), Maloyas (5%), and Berón de Astrada (3%) were Class III compatible to the S. mansoni. None of the 100 snails exposed from Caá-Catí became infected (Class 0 incompatible). Tested samples from Paraguay (Encarnación) were infected (20%) and compatible (Class III). It was also studied the persistence of the infection in 244 snails of the first generation (F1) of those that were susceptible from three places. It was demonstrated an increment of the susceptibility in the F1 from Maloyas (chi2 = 27.22; p = 0.0001) and Posadas (chi2 = 4.24; p = 0.04). The results point out the possibility that schistosomiasis might be able to spread into the Paraná River basin where B. tenagophila exists.

The subgenus Migonemyia Galati 1995 (Diptera, Psychodidae, Phlebotominae), with description of a new species Migonemyia vaniae: a review

The capture of a new species of the subgenus Migonemyia Galati, 1995 (Diptera, Psychodidae, Phlebotominae), Migonemyia vaniae sp. nov. in the Ribeira Valley, state of São Paulo, Brazil, together with the other two species: Mg. migonei (França, 1920) and Mg. rabelloi (Galati & Gomes, 1992) lead us to review this subgenus. The new species was described and illustrated. The genitalia of the two other species were also illustrated and some genital characteristics (number of setae on the gonocoxite tuft, ejaculatory ducts and pump and ducts/pump ratio; and number of setae on the tergite VIII of the females) considered important to differentiate the three species, including five populations of Mg. migonei (from Northeastern, Southeastern, and Southern Brazilian regions and of Peru) were submitted to variance analyses. The Mg. migonei population of Northeastern Brazilian region showed distinct smaller values (P < 0.05) than the other Brazilian populations studied as regarding these characteristics. The capture of both sexes of these three species in sympatry confirms the association between the sexes of Mg. rabelloi, recognised as doubtful when this species was originally described. Identification keys for male and female of the three species are presented.

Biology of three species of North American Triatominae (Hemiptera: Reduviidae: Triatominae) fed on rabbits

Aspects related to hatching, lifetime, number of blood meals for molting, mortality, feeding time, and postfeeding defecation delay were evaluated and compared in each instar of three North American Triatominae: Triatoma gerstaeckeri, Triatoma lecticularia and Triatoma protracta, all of them fed on rabbits. No significant differences (p > 0.05) were found among the three species regarding mean hatching rate, which was close to 20 days. Egg-to-adult development times were significantly shorter (p < 0.05) for T. lecticularia. Number of blood meals for molting to next instar ranged from one to five for T. protracta, and from one to six for T. gerstaeckeri and T. lecticularia. Mortality rates were higher in younger nymphs of T. lecticularia and T. protracta, while rates in T. gerstaeckeri were higher in fifth-instar nymphs. Mean feeding time was longest in T. gerstaeckeri, followed by T. lecticularia. More than twice the number of T. gerstaeckeri nymphs completed the development process, if compared to the nymphs from the other two species. Defecation delay was less than 10 min for T. lecticularia, T. protracta and the youngest nymphs of T. gerstaeckeri. Results point out that these three species may be important potential vectors of Trypanosoma cruzi for human populations, in areas of Mexico where these species are currently present.