Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata / Determinación de concentraciones letales y niveles de enzimas metabólicas de insecticidas en Triatoma dimidiata

Abstract: Objective: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. Materials and methods: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. Results: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and β-eterase levels. Conclusion: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

Resumen: Objetivo: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. Material y métodos: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. Resultados: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y β-eterasas. Conclusión: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

Where do these bugs come from? Phenotypic structure of Triatoma infestans populations after control interventions in the Argentine Chaco

House re-invasion by native triatomines after insecticide-based control campaigns represents a major threat for Chagas disease vector control. We conducted a longitudinal intervention study in a rural section (Area III, 407 houses) of Pampa del Indio, northeastern Argentina, and used wing geometric morphometry to compare pre-spray and post-spray (re-infestant bugs) Triatoma infestans populations. The community-wide spraying with pyrethroids reduced the prevalence of house infestation by T. infestans from 31.9% to < 1% during a four-year follow-up, unlike our previous studies in the neighbouring Area I. Two groups of bug collection sites differing in wing shape variables before interventions (including 221 adults from 11 domiciles) were used as a reference for assigning 44 post-spray adults. Wing shape variables from post-spray, high-density bug colonies and pre-spray groups were significantly different, suggesting that re-infestant insects had an external origin. Insects from one house differed strongly in wing shape variables from all other specimens. A further comparison between insects from both areas supported the existence of independent re-infestation processes within the same district. These results point to local heterogeneities in house re-infestation dynamics and emphasise the need to expand the geographic coverage of vector surveillance and control operations to the affected region.

Is imidacloprid an effective alternative for controlling pyrethroid-resistant populations of Triatoma infestans (Hemiptera: Reduviidae) in the Gran Chaco ecoregion?

The prevention of Chagas disease is based primarily on the chemical control of Triatoma infestans (Klug) using pyrethroid insecticides. However, high resistance levels, correlated with control failures, have been detected in Argentina and Bolivia. A previous study at our laboratory found that imidacloprid could serve as an alternative to pyrethroid insecticides. We studied the delayed toxicity of imidacloprid and the influence of the blood feeding condition of the insect on the toxicity of this insecticide; we also studied the effectiveness of various commercial imidacloprid formulations against a pyrethroid-resistant T. infestans population from the Gran Chaco ecoregion. Variations in the toxic effects of imidacloprid were not observed up to 72 h after exposure and were not found to depend on the blood feeding condition of susceptible and resistant individuals. Of the three different studied formulations of imidacloprid on glass and filter paper, only the spot-on formulation was effective. This formulation was applied to pigeons at doses of 1, 5, 20 and 40 mg/bird. The nymphs that fed on pigeons treated with 20 mg or 40 mg of the formulation showed a higher mortality rate than the control group one day and seven days post-treatment (p < 0.01). A spot-on formulation of imidacloprid was effective against pyrethroid-resistant T. infestans populations at the laboratory level.

Analysing deltamethrin susceptibility and pyrethroid esterase activity variations in sylvatic and domestic Triatoma infestans at the embryonic stage

The aim of the present work was to study the deltamethrin susceptibility of eggs from Triatoma infestans populations and the contribution of pyrethroid esterases to deltamethrin degradation. Insects were collected from sylvatic areas, including Veinte de Octubre and Kirus-Mayu (Bolivia) and from domiciliary areas, including El Palmar (Bolivia) and La Pista (Argentina). Deltamethrin susceptibility was determined by dose-response bioassays. Serial dilutions of deltamethrin (0.0005-1 mg/mL) were topically applied to 12-day-old eggs. Samples from El Palmar had the highest lethal dose ratio (LDR) value (44.90) compared to the susceptible reference strain (NFS), whereas the Veinte de Octubre samples had the lowest value (0.50). Pyrethroid esterases were evaluated using 7-coumaryl permethrate (7-CP) on individually homogenised eggs from each population and from NFS. The El Palmar and La Pista samples contained 40.11 and 36.64 pmol/min/mg protein, respectively, and these values were statistically similar to NFS (34.92 pmol/min/mg protein) and different from Kirus-Mayu and Veinte de Octubre (27.49 and 22.69 pmol/min/mg protein, respectively). The toxicological data indicate that the domestic populations were resistant to deltamethrin, but no statistical contribution of 7-CP esterases was observed. The sylvatic populations had similar LDR values to NFS, but lower 7-CP esterase activities. Moreover, this is the first study of the pyrethroid esterases on T. infestans eggs employing a specific substrate (7-CP).

Toxicological and biochemical analysis of the susceptibility of sylvatic Triatoma infestans from the Andean Valley of Bolivia to organophosphate insecticide

To increase our knowledge of the natural susceptibility of Triatoma infestans to an organophosphate insecticide, we performed toxicological and biochemical studies on three sylvatic populations from Bolivia and two populations from domestic dwellings from Bolivia and Argentina. Fifty-per-cent lethal doses (LD50) were determined based on the topical application of fenitrothion on first instar nymphs and mortality was assessed at 24 h. Both type of populations exhibited LD50ratios significantly higher than 1 with a range of the values (1.42-2.47); the maximum value were found in a sylvatic (-S) population, Veinte de Octubre-S. Samples were biochemically analysed using a glutathione S-transferase activity assay. The highest significant activity was obtained for Veinte de Octubre-S and the lowest activity was obtained for the reference population (102.69 and 54.23 pmol per minute per mg of protein respectively). Two out of the three sylvatic populations (Veinte de Octubre-S and Kirus Mayu-S) exhibited significantly higher glutathione S-transferase activity than that of the reference population. Based on this analysis of the natural susceptibility of this organism to organophosphate insecticides, continental and focal surveys of organophosphate susceptibility should be conducted to evaluate the evolution and distribution of this phenomenon.

Potential sources of Triatoma infestans reinfesting peridomiciles identified by morphological characterization in Los Llanos, La Rioja, Argentina

The presence of Triatoma infestans in habitats treated with insecticides constitutes a frequent problem in endemic areas. Basing our study on the hypothesis that descendants of a residual population should be more similar to the pre-treatment population than to any other, we compared the indications of two quantitative morphological approaches. This study seeks to find the origin of 247 T. infestans from three populations found in two chicken coops and a goat corral after treatment with insecticides. The results obtained by quantitative morphology suggest that the T. infestans found between three-34 months after the application of insecticides formed mixed populations with insects derived from residual foci and neighbouring habitats. Our analyses also showed the presence of a phenotype which does not resemble neither the pre-treatment phenotype nor the one from neighbouring populations, suggesting the presence of a particular post-treatment phenotype. The heads size showed some variations in males from different populations and remained unchanged in females, which reinforces the hypothesis of an intraspecific competition for food with priority for females. This article presents, for the first time, the combined analysis of geometric morphometry of heads and antennal phenotypes to identify the composition of reinfesting populations.

Response of Triatoma infestans to pour-on cypermethrin applied to chickens under laboratory conditions

This article reports the effects of a pour-on formulation of cypermethrin (6 percent active ingredient) applied to chickens exposed to Triatoma infestans, the main vector of Chagas disease in rural houses of the Gran Chaco Region of South America. This study was designed as a completely random experiment with three experimental groups and five replicates. Third instar nymphs were fed on chickens treated with 0, 1 and 2 cc of the formulation. Nymphs were allowed to feed on the chickens at different time intervals after the insecticide application. Third-instar nymphs fed on treated chickens showed a higher mortality, took less blood during feeding and had a lower moulting rate. The mortality rate was highest seven days after the insecticide solution application and blood intake was affected until 30 days after the application of the solution.

Toxicity of pyrethroids and repellency of diethyltoluamide in two deltamethrin-resistant colonies of Triatoma infestans Klug, 1834 (Hemiptera: Reduviidae)

The aim of the currrent investigation was to evaluate (a) the toxicity of three pyrethroids (deltamethrin, lambda-cyhalothrin, and tetramethrin); (b) the effect of these insecticides on the locomotor activity; and (c) the repellent effect of N,N-diethyl-m-toluamide (DEET) on two deltamethrin-resistant strains of Triatoma infestans from Argentina (El Chorro and La Toma), and one susceptible strain. The resistance ratios (RRs) obtained for the La Toma strain were: > 10,769, 50.7, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The RRs for the El Chorro strain were: > 10,769, 85.8, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The hyperactivity usually caused by the three pyrethroids was in both the deltamethrin-resistant strains compared to the susceptible reference strain. No differences were observed in the repellent effect of DEET between the three groups. These results indicate that the deltamethrin-resistant insects have a cross resistance to lambda-cyhalothrin and tetramethrin, and are also resistant to the first symptom of pyrethroid poisoning (hyperactivity). However, the sensorial process related to DEET repellency does not appear to be altered.

Ticks, ivermectin, and experimental Chagas disease

Following an infestation of dogticks in kennels housing dogs used for long-term studies of the pathogenesis of Chagas disease, we examined the effect of ivermectin treatment on the dogs, ticks, trypanosome parasites, and also on triatomine vectors of Chagas disease. Ivermectin treatment was highly effective in eliminating the ticks, but showed no apparent effect on the dogs nor on their trypanosome infection. Triatominae fed on the dogs soon after ivermectin treatment showed high mortality, but this effect quickly declined for bugs fed at successive intervals after treatment. In conclusion, although ivermectin treatment may have a transient effect on peridomestic populations of Triatominae, it is not the treatment of choice for this situation. The study also showed that although the dogticks could become infected with Trypanosoma cruzi, this only occurred when feeding on dogs in the acute phase of infection, and there was no evidence of subsequent parasite development in the ticks.

Biological effect of 1-dodecanol in teneral and post-teneral Rhodnius prolixus and Triatoma infestans (Hemiptera: Reduviidae)

Topical application of 1-dodecanol was significantly more toxic against teneral first nymphs (1-3 h old) than post-teneral first nymphs (24 h old). The lethal dose ratios were 711,500 for Rhodnius prolixus and 3613 for Triatoma infestans. No significative difference between LD50 was found when 1-dodecanol was injected in recently hatched adult R. prolixus (1-4 h old) nor in older adults (24 h old). These values were similar to those calculated for deltamethrin (an effective triatomicide), showing that 1-dodecanol had no insecticidal properties when it was applied by injection. Topical application of high dose of 1-dodecanol (1 æg/i) on teneral first nymphs of R. prolixus, produced an interruption of the darkening process of the cuticle, and probably in the development of its physiological properties.

Comparison of intervention strategies for control of Triatoma dimidiata in Nicaragua

The effectiveness of three operational strategies for the control of Triatoma dimidiata was compared by a field trial in the Department of Madriz, Nicaragua. One strategy involved full pretrial evaluation, followed by spraying of all houses irrespective of whether or not they had been found to be infested. The second strategy minimised the pretrial evaluation by considering the locality infested as soon as one house was found to be positive, followed by spraying all houses. The third strategy involved full pretrial evaluation, followed by spraying only those houses found to be positive. Evaluation after twelve months indicated that all three strategies were similarly effective, since all sprayed houses remained free of infestation. However, comparative estimates of the unit intervention costs indicated that strategies 1 and 2 were substantially less efficient than the third strategy of spraying only positive houses

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

Susceptibility and resistance to insecticides of Chagas disease vector

Chemical control of Chagas disease vectors appears to be the best pratical way to reduce the incidence of the disease. DDT was intially tested in the 1950s for the campaigns of control of Chagas disease vectors. Its low level of effectiveness against triatomine caused the failure of these control actions. HCH was then introduced in the southem cone and Dieldrin in the north of Latinoamerica. Starting in the late 1960s anticholinesterasic organophosphorus and carbamate compounds were introduced in the control of Chagas vectors. The use of pyrethoriod compounds began in 1980. This family of insecticides is now the most important tool in triatomines control because of its favorable toxicological properties. Other types of insecticides also studied for Chagas vector control were the insect growth regulators and the antifeeding compounds. Because of the mode of action of these insecticides they are now considered just a potential complement of neurotoxic insecticides for integrated programmes of Triatomines control. Innovative formulations such as fumigant canister and insecticidal paints have been successfully developed in Latinoamerica with the World Health Organization support. Resistance to insecticides of triatomines is not a great problem in Chagas vectors. However, some resistant strains to pyrethroids have been found in Rhodnius prolixus from Venezuela and in Triatoma infestans from Brazil. Some cases of T. infestans incipient resistance to deltamethrin have been detected in Argentina. According to the control tools now available it is possible to expect the interruption of vector transmission of Chagas disease in the near future.

Induction of glutathione S-transferase activity in Triatoma infestans

Several synthetic pesticides and allelochemicals used to treat Triatoma infestans adults by topic application showed some degree to cytosolic glutathione S-transferase (GST) induction. General inducers of detoxication systems such as phenobarbital and 3-methylcholantrene topically applied on T. infestans resulted in no GST induction. Meanwhile, general insecticide synergist such as piperonyl butoxide (160 µg/insect) increased the GST-activity in the range of 120-140 per cent. Insects injected with reduced glutathione (300 mg/insect) presented at the forth day elevated GST activity.

Incoordination, paralysis and recovery after pyrethoid treatment on nymphs III of Triatoma infestans (Hemiptera: reduviidae)

Symptons of poisoning for deltamethrin and cis-permethrin on nymphs III of Triatoma infestans were described. The time required for incoordination and paralysis were determined. Deltamethrin was equal or more rapid in the onset of the first (accordingly to dose), and cis-permethrin in the onset of the second one. There were no significant differences between incoordination doses 50 per cent (IncD50s) at different times for the two pyrethroids. They showed equivalent incoordination power, but the nymphs treated with deltamethrin recovered slighly more rapid and in greater amount than the nymphs treated with cis-permethrin. The recovery was inhibited by the simultaneus application of piperonyl butoxide. This suggests that biotransformation by mixed-function microsomal oxidases are involved in the process of recovery.

Chagas disease vector control in Tupiza, Southern Bolivia

Heavy domestic and peridomestic infestations of Triatoma infestans were controlled in two villages in southern Bolivia by the application of deltramethrin SC25 (2,5 per cent suspension concentrate) at a target dose of 25 mg a.i.m². Actual applied dose was monitored by HPLC analysis of filter papers placed at various heights on the house walls, and was shown to range from 0 to 59.6 about a mean of 28.5 mg a.i./m². Wall bioassays showed high mortality of T.infestans during the first month after the application of deltamethrin. Mortality declined to zero as summer temperatures increased, but reappeared with the onset of the following winter. In contrast, knockdown was apparent throughtout the trial, showing no discernible temperature dependence. House infestation rates, measured by manual sampling and use of paper sheets to collect bug faeces, declined from 79 per cent at the beginning of the trial to zero at the 6 month evalution. All but one of the houses were still free of T.infestans at the final evaluation 12 months after spraying, although a small number of bugs were found at this time in 5 of 355 peridomestic dependencies. Comparative cist studies endorse the recommendation of large-scale application of deltamethrin or pyrethroid of similar cost-effectiveness, as a means to eliminate domestic T.infestans populations in order to interrupt transmission of Chagas disease.

Evaluación de la B-cipermetrina para el control de Triatoma infestans / Evaluation of -cypermethrin for control of Triatoma infestans

This study assessed the insecticidal effect in the laboratory and in the field of the new pyrethroid -cypermethrin against Triatoma infestans, the vector of Chagas' disease, and compared it with that of deltamethrin. Comparison of the 50% lethal dosis (LD50) of both pyrethroids showed that -cypermethrin is more effective against the nymphs and that deltamethrin is more effective against the adults. Evaluation of the 50% lethal concentration (LC50) of the flowable formulations of both insecticides, placed on different surfaces, showed that their effectiveness on glass is similar, while on a ceramic surface deltamethrin is slightly more effective. The flowable formulations of the two insecticides were tested in 100 infested homes in the province of Santiago del Estero, Argentina. Deltamethrin was applied at a surface concentration of 25 mg/m2 and -cypermethrin at 50 mg/m2. The real surface concentrations of these products were analyzed from filter paper samples placed on walls and ceilings. Gas chromatography revealed good agreement with the target concentrations. In addition, entomological assessments were carried out 60, 90, 180, and 365 days after the treatments. Infestations were found only during the last assessment: in the peridomiciliary area of 10% of the houses treated with -cypermethrin and in 7% of the houses treated with deltamethrin (6% in the peridomiciliary area and 1% inside the house). The results indicate that -cypermethrin's effectiveness in controlling T. infestans when applied at a concentration of 50 mg/m2 is similar to that of deltamethrin applied at a concentration of 25 mg/m2

En este estudio se evaluó en el laboratorio y en el campo el efecto insecticida sobre Triatoma infestans, vector de la enfermedad de Chagas, del nuevo piretroide ß-cipermetrina, y se comparó con el de la deltametrina. La comparación de las dosis letales 50 (DL50) de ambos piretroides indicó que la ß-cipermetrina es más efectiva en ninfas y la deltametrina, en adultos. La evaluación de la concentración letal 50 (CL50) de las dos formulaciones floables de ambos insecticidas depositados sobre distintos soportes mostró que su efectividad en vidrio es similar. En cerámica, la deltametrina es ligeramente más efectiva. Los dos insecticidas floables se evaluaron en 100 viviendas infestadas de la provincia de Santiago del Estero, Argentina. La deltametrina floable se aplicó a una concentración superficial de 25 mg/m2 y la ß-cipermetrina, de 50 mg/m2. Las concentraciones superficiales reales de ambos productos se analizaron por medio de papeles de filtro colocados en paredes y techos. El análisis por cromatografía de gases mostró buena coincidencia con las concentraciones previstas. Asimismo, se realizaron evaluaciones entomológicas 60, 90, 180 y 365 días después de los tratamientos. Solo en la última evaluación se encontró 10% de infestación en peridomicilios de viviendas tratadas con ß-cipermetrina y 7% en las viviendas tratadas con deltametrina (6% en el peridomicilio y 1% en el domicilio). Los resultados indican que la efectividad de la ß-cipermetrina para controlar a T. infestans cuando se aplica a una concentración de 50 mg/m2 es similar a la de la deltametrina cuando esta se administra a una concentración de 25 mg/m2

Estado actual de la seroprevalencia de la infección chagásica humana y canina en la comuna de San Pedro de Atacama, II Región de Antofagasta en Chile 1995 / Present status of seroprevalence of canine and human chagasic infection in San Pedro de Atacama, II Región, Antofagasta, Chile 1995

In order to assess the impact of a control program against triatoma infestans launched in 1988, based on insecticide spraying of dwellings, a serological survey for chagasic infection was carried out during 1995 in three localities from San Pedro de Atacama county (22º55' south lat., 68º12' west long.), II Region of Antofagasta in northern Chile. Blood samples from 531 children and adolescents and 65 dogs were subjected to ELISA test and indirect inmunofluorescent test for Chagas's disease respectively. Tests resulted positive in 12 (2,3 percent) persons, all above 5 years old, in contrast with the 16,8 percent serological positivity observed in 1985. Three (4,6 percent) dogs (two 0-12 months old) resulted positive. These results indicate that dwellings sprayings with long-term activity insecticides against T. infestans is a good tool to prevent new human infections with T. cruzi. However, active vector transmissiom among domestic animals (canines) could be recently acquired.

Epidemiología de la enfermedad de Chagas en Chile: seguimiento serológico de 1.906 habitantes de un área rural endémica de la IV región, 1991-1993 / Epidemiology of Chagas' disease in Chile: serological follow-up of 1.906 inhabitants from an endemic rural area in IV región, 1991-1993

The IV region (29º 30' -32º 13' south lat.) is located in the center of the geographical area of distribution of Chagas' disease in Chile. Triatoma infestans is the main and almost exclusive vector of trypanpsoma cruzi in this country. The mean prevalence rate of T. cruzi human infection in urban and periurban sections of IV region is 24,7 percent. To assess the impact of anti-T. infestans activities, by means of health education and sprayings of dwellings with insecticides, carried out in the IV Region since 1980, during january-february (summer) a serological follow-up to residents from 46 rural chagasic localities was performed. An indirect hemagglutination test and an indirect immunofluorescence test were done to each of the surveyed persons. In 1991, 303 (15.9 percent) out of 1,906 examined people resulted serologically positive. In 1992, previous discarding the positive individuals found in 1991. 1,334 persons were examined resulting positive 9 (0.7 percent). In 1993, 1,398 persons were surveyed and 26 (1.9 percent) were positive. It is neteworthy that none of these 35 positive persons had been surveyed in 1991, being difficult to assert if any was positive before. Two infants, daughters of positive mothers, serologically positive at the beginning, changed to negative in the following survey, indicatins that it was passive transfer of maternal specific antibodies. According to the results of this study, it is concluded that dwelling spraying with persistent-activity insecticides against T. infestans and health education are good tools in the control of T. cruzi human infection, particularly when the involved community participates