Long-term reduction of Trypanosoma cruzi infection in sylvatic mammals following deforestation and sustained vector surveillance in northwestern Argentina

Long-term variations in the dynamics and intensity of sylvatic transmission of Trypanosoma cruzi were investigated around eight rural villages in the semiarid Argentine Chaco in 2002-2004 and compared to data collected locally in 1984-1991. Of 501 wild mammals from 13 identified species examined by xenodiagnosis, only 3 (7.9%) of 38 Didelphis albiventris opossums and 1 (1.1%) of 91 Conepatus chinga skunks were infected by T. cruzi. The period prevalence in opossums was four-fold lower in 2002-2004 than in 1984-1991 (32-36%). The infection prevalence of skunks also decreased five-fold from 4.1-5.6% in 1984-1991 to 1.1% in 2002-2004. Infection in opossums increased with age and from summer to spring in both study periods. The force of infection per 100 opossum-months after weaning declined more than six-fold from 8.2 in 1988-1991 to 1.2 in 2002-2004. Opossums were mainly infected by T. cruzi lineage I and secondarily by lineage IId in 1984-1991, and only by T. cruzi I in 2002-2004; skunks were infected by T. cruzi IId in 1984-1991 and by IIc in 2002-2004. The striking decline of T. cruzi infection in opossums and skunks occurred in parallel to community-wide insecticide spraying followed by selective sprays leading to very low densities of infected Triatoma infestans in domestic and peridomestic habitats since 1992; to massive deforestation around one of the villages or selective extraction of older trees, and apparent reductions in opossum abundance jointly with increases in foxes and skunks. These factors may underlie the dramatic decrease of T. cruzi infection in wild reservoir hosts.     

Incidence of trypanosoma cruzi infection among children following domestic reinfestation after insecticide spraying in rural northwestern Argentina

Following increasing reinfestation with Triatoma infestans after insecticide spraying, the household incidence of infection with Trypanosoma cruzi in children was positively related to the domestic abundance of infected T. infestans and the presence or proportion of infected dogs or cats in Amamá, a rural village in northwestern Argentina. Seven (12.1%) children seronegative for antibodies to T. cruzi at baseline, with no history of travel or blood transfusion, seroconverted after three years. Six incident cases lived in houses heavily infested with T. infestans, with high proportions of bugs infected with T. cruzi and having fed on humans or dogs. The remaining incident case occurred under a very light domestic infestation detected only at the endpoint, and most bugs had fed on humans. Dogs had a 17 times greater force of infection than children (4.3% per year). Sustained vector surveillance is crucially needed in high-risk areas for Chagas disease such as the Gran Chaco.     

Trypanosoma cruzi infection in dogs and cats and household seroreactivity to T. cruzi in a rural community in northeast Brazil.

The prevalence of Trypanosoma cruzi parasitemia as determined by xenodiagnosis on domestic dogs and cats was correlated with household rates of seroreactivity to T. cruzi and household Panstrongylus megistus infestation in a rural area in northeast Brazil where P. megistus was the only domiciliary triatomine vector. T. cruzi infection was present in about 18% of domestic dogs and cats. Two-thirds of seroreactive children below age 10 resided in houses with T. cruzi-infected animals. In houses with a T. cruzi-infected dog or cat, as well as at least one infected P. megistus, the household rate of seroreactivity to T. cruzi was five times greater than in houses with non-infected domestic animals and no infected triatomine vectors. Domestic dogs and cats are important reservoirs of T. cruzi in an endemic area where P. megistus is the only domiciliary triatomine vector.     

Infectivity to the vector Triatoma infestans of dogs infected with Trypanosoma cruzi in north-west Argentina.

The influence of reinfections on the infectivity to Triatoma infestans of dogs seropositive to Trypanosoma cruzi was investigated in Amamá, north-west Argentina, before (1984) and after (1986-1987) elimination of domestic bugs with residual insecticides in 1985. Infectivity was quantified by the percentage of infected bugs in each xenodiagnosis of a seropositive dog, i.e. the infective force (IF). Exposure to reinfection was represented by the domiciliary density of infected bugs and by an index of the potentially-infective contacts (IPIC) experienced by each dog (calculated as the product of the relative density of bugs per dog, the proportion of infected bugs, and the proportion of bugs taking blood meals from the dog). IF was unrelated to the dog's age or sex, or the level of specific antibodies to T. cruzi. Before vector elimination, the IF of seropositive dogs was significantly associated with both the density of infected bugs and the IPIC. After vector elimination, seropositive dogs resident at houses which had 10-70 infected bugs before spraying showed a higher median IF (67-75%) than those living in houses with zero to eight infected bugs before spraying (47-55%). The age-specific IF of seropositive dogs aged one year or older, recorded before (45-64%) and after (52-56%) the elimination of domestic bugs, did not differ significantly. Our data suggest that the infectivity of seropositive dogs to bugs is modified by previous or current exposure to bug-mediated reinfections, and that this relationship might be irreversible in the short term.     

Impact of community-based vector control on house infestation and Trypanosoma cruzi infection in Triatoma infestans, dogs and cats in the Argentine Chaco

The relative impact of two community-based vector control strategies on house infestation by Triatoma infestans and Trypanosoma cruzi infection in bugs, domestic dogs and cats was assessed in two neighboring rural areas comprising 40 small villages and 323 houses in one of the regions most endemic for Chagas disease in northern Argentina. The prevalence and abundance of domestic infestation were 1.5- and 6.5-fold higher, respectively, in the area under pulsed, non-supervised control actions operating under the guidelines of the National Vector Control Program (NCVP) than in the area under sustained, supervised surveillance carried out jointly by the UBA research team and NCVP. The prevalence of infestation and infection varied widely among village groups within each area. In the pulsed control area, the prevalence of infection in bugs, dogs and cats was two- to three-fold higher than in the area under sustained surveillance, most of the infected animals qualified as autochthonous cases, and evidence of recent transmission was observed. Infection was highly aggregated at the household level and fell close to the 80/20 rule. Using multiple logistic regression analysis clustered by household, infection in dogs was associated positively and significantly with variables reflecting local exposure to infected T. infestans, thus demonstrating weak performance of the vector surveillance system. For high-risk areas in the Gran Chaco region, interruption of vector-mediated domestic transmission of T. cruzi requires residual insecticide spraying that is more intense, of a higher quality and sustained in time, combined with community participation and environmental management measures.     

Detection of Trypanosoma cruzi infection in naturally infected dogs and cats using serological,parasitological and molecular methods

Domestic dogs and cats are major domestic reservoir hosts of Trypanosoma cruzi and a risk factor for parasite transmission. In this study we assessed the relative performance of a polymerase chain reaction assay targeted to minicircle DNA (kDNA-PCR) in reference to conventional serological tests, a rapid dipstick test and xenodiagnosis to detect T. cruzi infection in dogs and cats from an endemic rural area in northeastern Argentina. A total of 43 dogs and 13 cats seropositive for T. cruzi by an immunosorbent assay (ELISA) and an indirect hemagglutination assay (IHA), which had been examined by xenodiagnosis, were also tested by kDNA-PCR. kDNA-PCR was nearly as sensitive as xenodiagnosis for detecting T. cruzi-infectious dogs and cats. kDNA-PCR was slightly more sensitive than xenodiagnosis in seropositive dogs (91% versus 86%, respectively) and cats (77% against 54%, respectively), but failed to detect all of the seropositive individuals. ELISA and IHA detected all xenodiagnosis-positive dogs and both outcomes largely agreed (kappa coefficient, κ = 0.92), whereas both assays failed to detect all of the xenodiagnosis-positive cats and their agreement was moderate (κ = 0.68). In dogs, the sensitivity of the dipstick test was 95% and agreed closely with the outcome of conventional serological tests (κ = 0.82). The high sensitivity of kDNA-PCR to detect T. cruzi infections in naturally infected dogs and cats supports its application as a diagnostic tool complementary to serology and may replace the use of xenodiagnosis or hemoculture.     

A study of experimental reinfection by Trypanosoma cruzi in dogs.

The role of reinfection in the evolution of Chagas' disease was evaluated in dogs alternately infected with the 147 and SC-1 strains of Trypanosoma cruzi. A parasitologic, serologic, clinical, and electrocardiographic follow-up was carried out on the infected and noninfected dogs. The dogs were reinfected five times over a period of 38 months. No deaths were observed during the experiment. They presented a brief oligosymptomatic acute phase. The level of parasitemia decreased progressively with the number of reinfections. Bloodstream parasites were not detectable after the fifth reinfection. All parasite samples isolated during the follow-up were zymodeme B, corresponding to strain 147, irrespective of the strain with which the dogs were first infected and of the triatomine species used for isolation. Conversely, amplification by the polymerase chain reaction of a segment of the T. cruzi mini-exon gene showed the simultaneous presence of both strains in three of the eight reinfected animals. Antibody titers were greater among the dogs successively infected than those infected only once. Neither amastigotes nor T. cruzi DNA were detected in the tissues of the infected dogs. Alterations related to Chagas' disease were identified only in the heart and consisted of chronic focal and discrete myocarditis, compatible with the indeterminate form of Chagas' disease. All infected dogs developed this form of the disease, which was independent of the number of infections.     

Long-term effects of a nationwide control program on the seropositivity for Trypanosoma cruzi infection in young men from Argentina

Unselected nationwide cohorts of Argentine men 18 years of age summoned for military service were tested for antibodies to Trypanosoma cruzi each year from 1981 to 1993. After an initial screening using indirect hemagglutination test, the positive sera were retested by titrated indirect hemagglutination and immunofluorescence antibody tests at 39 laboratories or at the national reference center in Buenos Aires. Nearly 1.8 million men were examined for T. cruzi antibodies using the same standardized procedures under a quality assurance program. The prevalence of seropositivity for T. cruzi decreased significantly from 5.8% in 1981 to 1.9% in 1993, but the decrease was not homogeneous among provinces within each region or constant over time. Prior to the nationwide control campaign initiated in 1961-1962, 15 provinces had high (> 20%) percentages of houses with domiciliary infestation by Triatoma infestans bugs, which decreased to nine provinces in 1982, and four provinces in 1992. The observed decrease in the prevalence of seropositivity for T. cruzi may be mostly attributed to the spraying with insecticides to eliminate the domiciliary populations of Triatoma infestans. The lack of a sustainable triatomine surveillance program set a limit to the decrease of seropositivity rates and prompted a revised strategy based on community participation.     

Evaluation of recombinant antigens of Trypanosoma cruzi to diagnose infection in naturally infected dogs from Chaco region,Argentina

Dogs are considered the main mammal reservoir of Trypanosoma cruzi in domiciliary environments. Consequently, accurate detection of T. cruzi infection in canine populations is epidemiologically relevant. Here, we analysed the utility of the T. cruzi recombinant antigens FRA, SAPA, CP1, Ag1 and a SAPA/TSSA VI mixture, in an ELISA format. We used a positive control group of sera obtained from 38 dogs from the Chaco region in Argentina with positive homogenate‐ELISA reaction, all of them also positive by xenodiagnosis and/or PCR. The negative group included 19 dogs from a nonendemic area. Sensitivity, specificity, area under the curve (AUC) of the receiver operating charactheristic (ROC) curve and Kappa index were obtained to compare the diagnostic efficiency of the tests. The SAPA/TSSA VI had the highest performance, with a sensitivity of 94·7% and an AUC ROC of 0·99 that indicates high accuracy. Among individual antigens, SAPA‐ELISA yielded the highest sensitivity (86·8%) and AUC ROC (0·96), whereas FRA‐ELISA was the least efficient test (sensitivity = 36·8%; AUC ROC = 0·53). Our results showed that the use of SAPA/TSSA VI in ELISAs could be a useful tool to study dogs naturally infected with T. cruzi in endemic areas.     

Natural Trypanosoma cruzi infection in dogs of endemic areas of the Argentine Republic

The population dynamics and the prevalence of chagasic infection of 352 dogs living in 108 rural houses infested by triatomines were studied. The region was divided into three sections according to increasing distances to an urban area. Each animal was identified by means of its particular characteristics and built, and its owners gave information about its habits. By means of xenodiagnosis, serology and ECG studies, prevalences of infection, parasitological-serological correlation, percentage of altered electrocardiographic outlines and percentage of houses with parasitemic dogs, were determined. The rural area showed a characteristic T. cruzi infection pattern and differences in the canine population parameters with respect to the other areas were observed: a higher proportion of puppies than adult dogs, a more sedentary population, higher prevalences of infection, as measured by xenodiagnosis, in dogs, and the highest proportion of bedroom insects infected with T. cruzi. It is assumed that the sedentary characteristics of the human population in that rural area impinge in the blood offer to the triatomine population, and the high percentage of parasitemic dogs of the area, contribute to the rise of "kissing bugs" infected with T. cruzi found in bedrooms.     

The sylvatic transmission cycle of Trypanosoma cruzi in a rural area in the humid Chaco of Argentina

Little is known about the sylvatic transmission cycle of Trypanosoma cruzi in the Gran Chaco ecoregion. We conducted surveys to identify the main sylvatic hosts of T. cruzi, parasite discrete typing units and vector species involved in Pampa del Indio, a rural area in the humid Argentinean Chaco. A total of 44 mammals from 14 species were captured and examined for infection by xenodiagnosis and polymerase chain reaction amplification of the hyper-variable region of kinetoplast DNA minicircles of T. cruzi (kDNA-PCR). Ten (22.7%) mammals were positive by xenodiagnosis or kDNA-PCR. Four of 11 (36%) Didelphis albiventris (white-eared opossums) and six of nine (67%) Dasypus novemcinctus (nine-banded armadillos) were positive by xenodiagnosis and or kDNA-PCR. Rodents, other armadillo species, felids, crab-eating raccoons, hares and rabbits were not infected. Positive animals were highly infectious to the bugs that fed upon them as determined by xenodiagnosis. All positive opossums were infected with T. cruzi I and all positive nine-banded armadillos with T. cruzi III. Extensive searches in sylvatic habitats using 718 Noireau trap-nights only yielded Triatoma sordida whereas no bug was collected in 26 light-trap nights. Four armadillos or opossums fitted with a spool-and-line device were successfully tracked to their refuges; only one Panstrongylus geniculatus was found in an armadillo burrow. No sylvatic triatomine was infected with T. cruzi by microscopical examination or kDNA-PCR. Our results indicate that two independent sylvatic transmission cycles of T. cruzi occur in the humid Chaco. The putative vectors of both cycles need to be identified conclusively.     

In vivo and in vitro effects of cyclosporin A on Trypanosoma cruzi

The in vivo and in vitro effects of cyclosporin A on T. cruzi were examined. Mice receiving 150 or 75 mg/kg/day of cyclosporin A and infected with T. cruzi 48 hr later had significantly higher parasitemias and earlier mortality than controls. Mice receiving cyclosporin A after infection had parasitemias similar to controls. Infections with both the reticulotropic Y and the miotropic CL strains of T. cruzi were enhanced by cyclosporin A. The in vitro replication of epimastigotes, but not the intracellular replication of amastigotes, in mouse macrophages was inhibited by 5 micrograms cyclosporin A/ml. Enhancement of the infection by cyclosporin A was not due to an effect on macrophages since the drug did not prevent development of activated macrophages capable of killing intracellular T. cruzi.     

Infected dogs as a risk factor in the transmission of human Trypanosoma cruzi infection in western Venezuela

A total of 565 mongrel dogs from rural localities of Venezuela were examined by serological (DAT, IFAT and ELISA) and parasitological tests to address the status of Trypanosoma cruzi infection and to evaluate their role in the transmission of the infection to human population. The overall percentage of sero-positive infected dogs shown to be 67.6% (382/565):253 (61.7%) from 47 villages belonging to 8 states located at 4 different geographical regions of western Venezuela and 129 (33.5%) dogs from 48 households located in areas where Chagas disease is endemic. From 101 sampled dogs living in close proximity to 30 acute chagasic patients, 84% expressed specific anti-T. cruzi antibodies (Ab) with 12 of them (14%) showing blood circulating parasites (BCP). In these houses a high proportion of sero-positive people (20%) and frequent indoor infestation by triatomine-bugs (70%) was also recorded. The analysis revealed that from the 47 rural villages sampled during the study, 91.5% had the presence of T. cruzi sero-positive dogs, ranging from 62% positive localities at the states of Falcon and Cojedes to 100% in the other six studied Venezuelan states. This demonstrates that T. cruzi-infected dogs are found throughout all the geographical regions of western Venezuela irrespective of their ecological differences. Molecular typing of T. cruzi isolates from infected dogs using ribosomal and mini-exon gene markers, revealed the presence of both T. cruzi I and T. cruzi II lineages. The coincidence in the circulation of T. cruzi II in dog and human populations at the same locality and at the same time is reported and its significance is discussed. The combined serological, parasitological, epidemiological and molecular data is gathered here to call the attention on the presence of infected dogs as a risk factor in the maintenance of T. cruzi as a source for infection to humans.     

Exacerbation of Trypanosoma cruzi infection in mice treated with the immunoregulatory agent cyclosporin A

Treatment with the immunoregulatory fungus metabolite cyclosporin A exacerbated the course of Trypanosoma cruzi infection in outbred as well as in inbred BALB/c mice. This effect was not observed in congenitally athymic, nu/nu, mice but was readily reproduced in their thymus-bearing, nu/+, littermates. These results suggest that the deleterious action of cyclosporin A on the course of experimental Chagas' disease results from activity of the drug on thymus or thymus-derived lymphocytes and emphasizes the role of T cells in host defense against T. cruzi infection.     

New Sylvatic Hosts of Trypanosoma cruzi and Their Reservoir Competence in the Humid Chaco of Argentina: A Longitudinal Study

A four-year longitudinal study of the structure of sylvatic transmission cycles of Trypanosoma cruzi, reservoir host competence and parasite discrete typing units was conducted in a disturbed rural area of the humid Chaco in Argentina. Among 190 mammals examined by xenodiagnosis and polymerase chain reaction amplification, the composite prevalence of infection was substantially higher in Dasypus novemcinctus armadillos (57.7%) and Didelphis albiventris opossums (38.1%) than in Euphractus sexcinctus (20.0%), Tolypeutes matacus (12.5%), and Chaetophractus vellerosus (6.3%) armadillos. Trypanosoma cruzi was detected for the first time in Thylamys pusilla small opossums and in two unidentified small rodents. Infection was spatially aggregated only in armadillos. All Didelphis were infected with T. cruzi I and all armadillo species were infected with T. cruzi III, implying two distinct sylvatic cycles with no inputs from the domestic cycle. Dasypus armadillos and Didelphis opossums were much more infectious to vectors than other armadillos, small opossums, or rodents.     

Differential impact of metacyclic and blood trypomastigotes on parasitological, serological and phenotypic features triggered during acute Trypanosoma cruzi infection in dogs

A detailed follow-up investigation of the major parasitological, serological and phenotypic features in dogs experimentally infected with metacyclic (MT) and blood (BT) trypomastigotes of Trypanosoma cruzi strain Berenice-78, typifying vectorial and transfusional transmission of human Chagas disease, has been conducted. Although there were no changes with respect to the window of patent-parasitaemia, significant differences between MT- and BT-infected dogs in both the prepatent period (days 23 and 19, respectively) and the day of maximum parasitaemia (days 26 and 22, respectively) were recorded. A progressive enhancement in the level of T. cruzi-specific antibodies accompanied infection by both MT and BT forms, although higher IgG titres developed on days 14 and 21 following infection with MT forms. Higher Thy-1(+)/CD21(+) and lower CD4(+)/CD8(+) cell ratios, occasioned by increased levels of Thy-1(+) and CD8(+) T-cells and reduced frequencies of CD4(+) T-cells and CD21(+) B-lymphocytes, were observed in both MT- and BT-infected animals. The reduced frequency of CD14(+) leukocytes was revealed as the most relevant phenotypic feature intrinsic to T. cruzi infection independent of inoculum source. BT-specific phenotypic features included an early reduction in the percentage of circulating CD21(+) and CD14(+) leukocytes, together with a higher Thy-1(+)/CD21(+) cell ratio on day 42. On the other hand, higher levels of CD8(+) T-cells, together with a lower CD4(+)/CD8(+) cell ratio on day 28, were characteristic of MT infection. These findings emphasise the importance of inoculum source and suggest that vectorial or transfusional routes of T. cruzi infection may trigger distinct parasite-host interactions during acute Chagas disease.     

Seroprevalence and risk factors for Trypanosoma cruzi infection in the Amazon region of Ecuador

Trypanosoma cruzi infection in the Ecuadorian Amazon region has recently been reported. A seroepidemiologic survey conducted in four provinces in this region indicates a seroprevalence rate of 2.4% among the 6,866 samples collected in 162 communities. Among children < OR = 10 years of age, 1.2% were seropositive. Risk factors for T. cruzi seropositivity were having been born and remaining in the Ecuadorian Amazon provinces, age, living in a house with a thatch roof and open or mixed wall construction, recognizing the vector insects, and reporting being bitten by a triatomine bug. These data suggest active transmission of Chagas' disease in the Ecuadorian Amazon region is associated with poor housing conditions, and highlight the need for further studies aimed at understanding the biology of the insect vectors, reservoir species, and the clinical impact of T. cruzi infection as the basis for future educational and control programs in this region.     

Effects of partial housing improvement and insecticide spraying on the reinfestation dynamics of Triatoma infestans in rural northwestern Argentina

The long-term effects on domiciliary reinfestation by Triatoma infestans of smoothing the plaster of indoor walls prior to insecticide application (in Amamá village) relative to only insecticide application (in Trinidad-Mercedes villages) were evaluated in rural northwestern Argentina from 1992 to 1997. All domestic and peridomestic areas of each house were sprayed with 2.5% suspension concentrate deltamethrin at 25 mg/m(2) in October 1992, and infestations were assessed by various methods every 6 months. Domiciliary infestation decreased from 72-88% in 1992 to 6-17% in late 1995, to increase moderately thereafter without returning to baseline rates. Peridomestic sites were the first in becoming reinfested, and reached more abundant T. infestans populations than domiciliary areas. Domiciliary infestation rates and bug abundances were not significantly different between communities during surveillance. Domiciliary infestation rates in well-plastered houses were very low (5-9%) and approximately stable until 1996, but in houses with regular or bad plaster they consistently increased from 5 to 19-21% in both communities. Logistic multiple regression analysis showed that the likelihood of domestic infestation assessed through householders' collections was significantly and positively associated with the occurrence of an infested peridomestic site in the respective house, the occurrence of high-density domestic infestations before interventions, and well-plastered walls in 1996. Combining insecticide spraying and partial improvement of walls controlled domestic infestations and transmission of Trypanosoma cruzi effectively, but was not sufficient to eliminate T. infestans from the study area or increase the effectiveness of careful chemical control.