A simple method to identify triatomine (Hemiptera: Reduviidae) feces in sensing devices used in vector surveillance programs.

We successfully applied the phenolphthalin (Kastle-Meyer) test used in forensic chemistry to distinguish between feces from triatomines and other domestic arthropods in sensing devices used for vector surveillance. All black or dark brown, but not white or yellow, fecal smears from laboratory-reared or field-collected Triatoma infestans Klug, Triatoma guasayana Wydgozinsky & Abalos, Triatoma sordida Ståhl (recently revalidated as Triatoma garciabesi Carcavallo, Cichero, Martínez, Prosen & Ronderos) tested positive, whereas dejecta from cockroaches and spiders, crickets, beetles, predatory bugs, and domestic flies tested negative. Black or dark brown dejecta from female Aedes aegypti L. and Cimex lectularius L. bedbugs also tested positive. In sellsing devices installed in bedrooms of 11 houses in Amamá, rural northwestern Argentina, where neither cimicid bedbugs nor argasid ticks had been found over the years, only 62% of the black or dark brown fecal smears attributed to triatomines by a skilled observer tested phenolphthalin-positive. After insecticidal spraying, when bedroom areas were not colonized by triatomines, only 33-40% of the black or dark brown fecal smears in sensor boxes attributed to triatomines by another skilled observer tested phenolphthalin-positive. Eleven (79%) ofthe 14 houses with dubious or nontypical triatomine feces tested phenolphthalin-positive at least once during 1993-1995. Our study introduces a low-cost, simple and effective procedure for the identification of triatomine feces. The test, as a helpful adjunct to sensing devices used in triatomine surveillance, will aid in the accurate detection of infestations and the determination of the need for insecticide application.

Trypanosoma cruzi infection in Triatoma infestans and other triatomines: long-term effects of a control program in rural northwestern Argentina.

The prevalence of Trypanosoma cruzi infection in Triatoma infestans, Triatoma guasayana, and Triatoma sordida was evaluated in Amamá and other neighboring rural villages in northwestern Argentina for five years after massive spraying with deltamethrin in 1992 and selective sprays thereafter. Local residents and expert staff collected triatomines in domiciliary and peridomestic sites. During 1993-1997, the prevalence of T. cruzi was 2.4% in 664 T. infestans, 0.7% in 268 T. guasayana, and 0.2% in 832 T. sordida. T. cruzi infection was more frequently detected in adult bugs and in triatomines collected at domiciliary sites. The infected T. guasayana and T. sordida were nymphs and adults, respectively, captured at peridomestic sites. The prevalence of T. cruzi infection in T. infestans decreased from 7.7% to 1.5% during the surveillance period, although that change was not statistically significant. Comparison of T. infestans infection rates before the control program and during surveillance showed a highly significant decrease from 49% to 4.6% in bedrooms, as well as a fall from 6% to 1.8% in peridomestic sites. Because of its infection with T. cruzi and frequent invasion of domiciliary areas and attacks on humans and dogs, T. guasayana appeared implicated as a putative secondary vector of T. cruzi in domestic and peridomestic sites during the surveillance period. T. sordida was the most abundant species, but it was strongly associated with chickens and showed little tendency to invade bedrooms.

Monitoring house reinfestation by vectors of Chagas disease: a comparative trial of detection methods during a four-year follow-up.

Domestic reinfestations by triatomine bugs were monitored after application of deltamethrin and apparent elimination of Triatoma infestans in Amama and other nearby rural villages, north-west Argentina, from 1992 to 1996. The five methods used were sensor boxes, sheets of pink typing-paper, timed manual catches by a skilled three-person team aided by a flushing-out agent, collections by house-dwellers, and knockdown using insecticide fumigant canisters. In bedrooms, house-dwellers collected T. infestans significantly more frequently than the flushing-out method, but the reverse occurred in peridomestic sites. Both methods and sensor boxes revealed the frequent invasion of adult Triatoma guasayana and T. infestans, but neither T. guasayana nor Triatoma sordida colonized bedroom areas in spite of their rising abundance in nearby peridomestic sites. Sensor boxes were significantly more sensitive than the matched paper-sheets in three of five cross-sectional surveys. On average, each box recorded 2.0-3.2 times more triatomine fecal smears than each paper sheet. The frequency of dejecta in sensor boxes correlated positively with the proportion of houses where T. infestans, T. guasayana or T. sordida were captured by any method in bedroom areas. Triatomine fecal smears in sensor boxes were the earliest and most frequent sign of domiciliary infestation, followed by dwellers' collections of adult bugs. Analyzing the data prospectively, we provide a quantitative, predictive understanding of detection methods and review the validity and interpretation of the different signs of infestation obtained. The most sensitive and cost-effective combination of detection methods for vector surveillance in domestic areas was the use of sensor boxes and house-dwellers collections.

Household prevalence of seropositivity for Trypanosoma cruzi in three rural villages in northwest Argentina: environmental, demographic, and entomologic associations.

Environmental, demographic, and entomologic variables were analyzed by logistic multiple regression analysis for their association with the likelihood of being seropositive for Trypanosoma cruzi in three highly infested rural villages of northwest Argentina. The prevalence of seropositivity for T. cruzi, as determined by the composite results of three serologic tests, was 34% among 338 persons in 1992. The strongest positive predictors of the adjusted odds of being infected were the household number of dogs, the density of T. cruzi-infected Triatoma infestans in bedroom areas, and each person's age. Dwellers from houses with roofs made completely or partly with a grass called simbol, or which used insecticides rudimentarily and nonsystematically, had a significantly lower odds of being seropositive for T. cruzi than residents from other types of dwellings. The adjusted odds of infection also increased with the number of T. cruzi-infected dogs or cats and the presence of chickens in bedroom areas. No significant effects on the adjusted odds of infection of a community-wide deltamethrin spraying carried out in one of the villages seven years before were detected. Socioeconomic indicators, such as domiciliary area, and numbers of corrals and livestock, were inversely related to being infected. Our study identified several manageable variables suitable for control actions, most of them not examined before in univariate or multivariate analyses. Environmental management based on low-cost housing with appropriate local materials and removal of domestic animals from domiciliary areas have a crucial role to play in the control of Chagas' disease in rural areas.

Immunodiagnosis of Trypanosoma cruzi (Chagas' disease) infection in naturally infected dogs.

This study reports on the standardization of an enzyme-linked immunosorbent assay (ELISA) for detecting specific antibodies anti-Trypanosoma cruzi in naturally infected dogs. Sera from 182 mongrel dogs of all ages residing in four rural villages in Santiago del Estero, Argentina, were collected in November 1994 and preserved in buffered neutral glycerin. All sera were tested by indirect hemagglutination test (IHAT), indirect immunofluorescence test (IFAT), and ELISA using the flagellar fraction of T. cruzi as antigen. Dog sera from an area without vectorial transmission were used to calculate ELISA specificity and cut-off value. Eighty-six percent of sera had concordant results for all tests. All sera reactive for IHAT and IFAT were also reactive for ELISA, except in one case. Sera tested by ELISA when diluted 1:200 allowed a clearer division between non-reactive and reactive sera than when 1:100 with greater agreement among serologic techniques. The specificity of ELISA was 96.2%. Among 34 adult dogs with a positive xenodiagnosis, sensitivity was 94% both for ELISA and IFAT. ELISA is the first choice for screening purposes and one of the pair of techniques recommended for diagnostic studies in dog populations.

Immunodiagnosis of Trypanosoma cruzi (Chagas' disease) infection in naturally infected dogs

This study reports on the standardization of an enzyme-linked immunosorbent assay (ELISA) for detecting specific antibodies anti-Trypanosoma cruzi in naturally infected dogs. Sera from 182 mongrel dogs of all ages residing in four rural villages in Santiago del Estero, Argentina, were collected in November 1994 and preserved in buffered neutral glycerin. All sera were tested by indirect hemagglutination test (IHAT), indirect immunofluorescence test (IFAT), and ELISA using the flagellar fraction of T. cruzi as antigen. Dog sera from an area without vectorial transmission were used to calculate ELISA specificity and cut-off-value. Eighty-six percent of sera had concordant results for all tests. All sera reactive for IHAT and IFAT were also rective for ELISA, except in one case. Sera tested by ELISA when diluted 1:200 allowed a clearer division between non-reative and reactive sera than when 1:100 with greater agreement among serologic techniques. The specificity of ELISA was 96,2 per cent. Among 34 adult dogs with a positive xenodiagnosis, sensitivity was 94 per cent both for ELISA and IFAT. ELISA is the first choice for screening purposes and one of the pair of techniques recommended for diagnostic studies in dog populations.

Evaluation of dogs as sentinels of the transmission of Trypanosoma cruzi in a rural area of north-western Argentina.

Dogs, the main domestic reservoirs of Trypanosoma cruzi in the Argentine chaco, may be useful as sentinels of vector-mediated transmission of Trypanosoma cruzi in control programmes if canine infections acquired by all other routes could be excluded. In the present study, in the province of Santiago del Estero, Argentina, the dog populations in the neighbouring villages of Amamá, Trinidad, Mercedes, Villa Matilde and Pampa Pozo were studied immediately before spraying of residual insecticides (in 1992 in Amamá, Trinidad and Mercedes and 1993-1994 in the other villages) and twice during a subsequent programme of triatomine surveillance (in 1994 and 1996). Overall seropositivity for Trypanosoma cruzi infection (i.e. the proportion of dogs found positive by at least two different serological tests in any one survey) steadily declined from 65% (54/83) in 1992 to 39% (70/182) in 1994 and 15% (36/237) in 1996. No seroconversion was detected between the 1994 and 1996 surveys in any dog aged > 2 years. However, among the dogs aged < 2 years (which were born after virtual elimination of Triatoma infestans from the villages), seropositivity fell from 15% (12/81) in 1994 to 5% (6/131) in 1996. Thirteen demographic, behavioural and entomological variables were analysed by logistic multiple-regression analysis for their association with seropositivity in dogs born locally during the surveillance programme. The total number of Triatoma guasayana collected in the bedroom areas of the owner's house, having a mother which was seropositive for Trypanosoma cruzi, and the number of seropositive dogs with which it cohabited were all found to be significant predictors of seropositivity in any dog. The results are the first indication that Triatoma guasayana is the vector responsible for the new cases of Trypanosoma cruzi infection seen in dogs in domestic or peridomestic sites during the surveillance programme.

Probability of infection with Trypanosoma cruzi of the vector Triatoma infestans fed on infected humans and dogs in northwest Argentina.

The probability that an uninfected Triatoma infestans would become infected with Trypanosoma cruzi after a single feed on people or dogs seropositive for T. cruzi was estimated in Amama, a rural village in northwest Argentina where transmission had resurged four years earlier. The prevalence of seropositivity for T. cruzi was 34.2% among 225 people tested, and 65.1% among 83 dogs tested. Parasitemia was detected by xenodiagnosis in 29.3% of 41 seropositive persons and in 85.3% of 34 seropositive dogs. Parasitemia decreased with age more sharply in seropositive people than in seropositive dogs. Seropositive humans infected 2.6% (95% confidence interval = 1.6%-3.6%) of 963 third or fourth instar nymphs fed once on them, whereas dogs infected 48.7% (44.7%-52.7%) of 610 nymphs. The probability of bug infection increased significantly with instar and was positively related to molting success. The infectivity to bugs of seropositive dogs was 12 times higher than that of seropositive children, and 100 times higher than that of seropositive adults. The weighted probability of infection of an uninfected bug fed randomly on any dog (0.3082) was about 50 times higher than that of bugs fed on any human (0.0062). Such differences in relative infectivity, combined with the relative host-feeding preference of domiciliary Triatoma infestans for dogs, reinforces the important role of domestic dogs as a risk factor for the domestic transmission of T. cruzi.

Detecting domestic vectors of Chagas disease: a comparative trial of six methods in north-west Argentina.

Six methods for detecting domestic infestations by triatomine bugs were compared in the rural community of Amamá, north-west Argentina. An average of three pairs (range, 2-5 pairs) of sensor boxes and sheets of pink typing-paper were tacked to the walls of human sleeping areas in 45 houses for 30 days and then inspected by a two-man team. Triatoma infestans bugs were collected in bedrooms by a different two-man team aided by a flushing-out agent both before and after application of sensing devices. Finally, knockdown collections of bugs after application of one insecticide fumigant canister per bedroom were also made. The proportion of houses with evidence of current domestic bug infestations that were detected by the various methods were as follows: sensor boxes (95.3%), reports of householders (88.4%), knockdown (87.8%), paper-sheets (86.0%), and flushing-out (69.8-76.7%). The detectability of infestations, irrespective of the method used, increased with the density of the bugs. At low or intermediate bug densities, individual sensor boxes were more sensitive than their matched paper-sheets, but at any bug density there were no significant differences between the pooled results for all the boxes and for all the paper-sheets in the house. On average, each sensor box recorded 2.25 times more triatomine faecal smears than its matched paper-sheet, and this relation increased with the density of bugs in the house. Both sensing devices were effective at monitoring unsuccessful attempts of peridomestic triatomine populations to colonize houses.