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.

Insecticidal activity of neem oil against Gyropsylla spegazziniana (Hemiptera: Psyllidae) nymphs on Paraguay tea seedlings / Atividade inseticida de óleo de nim contra ninfas de Gyropsylla spegazziniana (Hemiptera: Psyllidae) em mudas de erva-mate

Abstract Gyropsylla spegazziniana (Paraguay tea ampul) is one of the most important pests of Paraguay tea plants, and prohibition of synthetic insecticide use for control of this pest has led to the search for alternative methods. This laboratory study aimed to compare different control strategies for G. spegazziniana, utilizing a commercial neem seed oil product. Paraguay tea seedlings were treated with neem oil solution both pre- and post-infestation with 5th instar nymphs. The systemic action of neem oil was also evaluated by treating plant soil with the neem oil solution, followed by transfer of the insects to plants 24 h post-treatment. Spray treatments were effective against the pest, especially post-infestation (80% mortality), demonstrating the potential of neem oil for control of the Paraguay tea ampul. No significant effects were observed with respect to systemic activity.

Resumo Gyropsylla spegazziniana (ampola da erva-mate) é uma das principais pragas da erva-mate, tanto em áreas de plantio como em viveiros. A proibição do uso de inseticidas para o controle de pragas da cultura tem levado à busca de alternativas. Este trabalho visou avaliar comparativamente, em laboratório, estratégias de aplicação de um produto comercial à base de óleo de nim sobre G. spegazziniana, em solução a 5%. Os testes foram realizados em mudas de erva-mate, tratadas pré e pós-infestação com ninfas de 5º instar do inseto, para avaliação de contato. Também, avaliou-se a ação sistêmica do óleo de nim, aplicando-se a solução do produto no solo da muda de erva-mate, seguido da transferência dos insetos após 24 h. Verificou-se que os tratamentos via pulverização foram eficientes contra a praga, principalmente na aplicação pós-infestação (80% de mortalidade), comprovando o potencial inseticida do óleo de nim contra a ampola-da-erva-mate em mudas. Entretanto, em relação à atividade sistêmica, não foram observados efeitos consideráveis sobre a praga.

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.

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.

Repellent activity of DEET against Amblyomma cajennense (Acari: Ixodidae) nymphs submitted to different laboratory bioassays / Atividade repelente do DEET contra ninfas de Amblyomma cajennense (Acari: Ixodidae) em bioensaio laboratorial

This study was developed to evaluate the repellent activity of N,N-diethyl-3-methylbenzamide (DEET) against Amblyomma cajennense nymphs. Two repellent bioassays were compared and the effective concentration and repellent time were calculated. The fingertip test was accomplished to evaluate in vivo four concentrations of the compound (0.200; 0.100; 0.050 and 0.025 mg.cm-2) and the filter-paper bioassay to evaluate in vitro the two highest concentrations. The compound provided repellence higher than 90 percent in all concentrations and at least 95 percent repellency in the highest concentration over 5 hours. The effective concentration against 50 percent of tested nymphs (EC50) was 0.006 mg.cm-2 and the EC99 was 0.036 mg.cm-2. Those concentrations were lower than the ones obtained against other tick species, denoting the effectiveness of DEET against A. cajennense. The repellency time against 50 percent of the ticks (RT50) was 4.8 hours and the RT90 was 2.7 hours. Both bioassays were adequate to evaluate A. cajennense repellency and provided similar results; however the in vivo test is more appropriate to estimate the effective concentration and repellency time.

Este estudo foi conduzido com o objetivo de avaliar a atividade repelente do N,N-diethyl-3-methylbenzamide (DEET) sobre ninfas de Amblyomma cajennense. Dois bioensaios para a avaliação de repelência foram comparados e cálculos da concentração eficaz e do tempo de repelência foram realizados. Foram empregados o bioensaio da ponta do dedo, para avaliação in vivo de quatro concentações do químico (0,200; 0,100; 0,050 e 0,025 mg.cm-2) e o bioensaio do papel filtro, para a avaliação in vitro das duas concentrações mais altas. O composto conferiu mais de 90 por cento de repelência em todas as concentrações utilizadas e 95 por cento de repelência por mais de cinco horas na maior concentração. A concentração do composto efetiva contra 50 por cento das ninfas testadas (CE50) foi de 0,006 mg.cm-2 e a CE99 foi de 0,036 mg.cm-2. Estas concentrações são mais baixas do que as observadas em outras espécies de carrapatos, denotando a efetividade do princípio contra A. cajennense. O tempo de repelência de 50 por cento dos carrapatos (TR50) foi de 4,8 horas e o TR90 de 2,7 horas. Os dois bioensaios avaliados permitiram a observação de percentuais de repelência igualmente altos e se mostraram adequados para tal avaliação, sendo que o teste in vivo é mais indicado para cálculo da concentração eficaz e da duração da repelência.

Susceptibility of Triatoma infestans to deltamethrin in Rio Grande do Sul, Brazil

Strategies for controlling Chagas disease are based on spraying infested houses with pyrethroid insecticides. However, the intense use of these insecticides has promoted resistance of Triatoma infestans and, in Argentina, Bolivia and Southern Brazil, low levels of resistance have been reported. Due to the persistence of T. infestans in the state of Rio Grande do Sul (RS), we evaluated the occurrence of deltamethrin resistance in four strains from different municipalities in comparison to two susceptible strains from Brazil and one resistant strain from Bolivia. The results indicated the absence of resistance in T. infestans from RS.

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.

Laboratory evaluation of the effects of triflumuron on the development of Rhodnius prolixus nymph

Studies were carried out to evaluate the efficacy of the growth regulator, triflumuron (TFM) (Starycide® sc 480 Bayer), for disrupting the development of Rhodnius prolixus fifth-instar nymph by oral, topical or continuous treatment. All treatments were able to induce high levels of mortality, delay development and molt inhibition. Oral treatment induced molt inhibition in all insects that survived at doses of 0.25, 0.50 and 5.0 mg/mL of a blood meal. The highest levels of both mortality in 24 h and molt inhibition were always observed after topical treatment. The lowest doses needed to obtain considerable biological effects were always observed after continuous treatment. In this way, the highest levels of mortality within 30 days were detected after continuous treatment, which also induced an extended inter-molting period, a lower number of over-aged nymphs and the highest level of molting in nymphs that survived. Moreover, the effects of TFM on insects were often displayed in a dose response manner. These results indicate that TFM acts as a potent growth inhibitor of R. prolixus nymphs and has the potential to be used in integrated vector control programs against hematophagous triatomine species.

New synthetic precocenoids as potential insect control agents.

Ageratochromes or precocenes are known for their insect growth regulating (IGR) activity. The present investigation was taken up with an objective to look for the lead structure in these compounds which can be elaborated synthetically to obtain useful growth regulators for practical purposes. With this in mind, some variants of precocenes were synthesized in the laboratory and tested for their toxicity and growth regulating activity using red cotton bug Dysdercus koenigii as the test insect. Most of the precocenoids showed toxicity of various degree and metamorphic derangements to different extents. Adults emerging from treated nymphs could not complete the normal life span. Among the compounds tested 8-acetyl-7-hydroxy-5-methoxy-dimethylchromene (alloevodinol) was more toxic and also showed developmental defects at very low dose such as 0.5 mg l(-1)/nymph. Precocene II (6, 7-dimethoxy-2, 2-dimethylchromene) was used as the standard compound. It was the least toxic and showed effects at 30 mg l(-1)/nymph.

Juvenile hormone activity in Dysdercus cingulatus Fabr by juvenile hormone esterase inhibitor, OTFP.

Application of juvenile hormone esterase inhibitor 3-octylthio-1,1,1- trifluropropan-2-one (OTFP) to 5th instar nymphs and virgin females of D. cingulatus revealed the profound role played by juvenile hormone esterase (JHE) in metamorphosis and reproduction. The ability of OTFP to cause delay and the formation of malformed nymphs, suggests that inhibition of JHE in vivo maintains a higher than normal hemolymph JH titer. It is obvious that OTFP does inhibit in vivo JHE activity in late instar nymphs. Further, the application of JHE inhibitor, OTFP to virgin females demonstrates that substituted trifluropropanones can indirectly stimulate egg development by inhibiting JHE activity in virgin females.

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.

Effect of sub-lethal concentrations of permethrin on ovary activation in the predator Supputius cincticeps (Heteroptera: Pentatomidae)

Inseticidas podem causar mortalidade e efeitos deletérios em percevejos predadores. Por isso investigou-se o efeito de cinco concentrações de permetrina aplicadas em ninfas de terceiro estádio de Supputius cincticeps (Stål) (Heteroptera: Pentatomidae) em sua ativação ovariana. Essas ninfas receberam aplicação tópica de permetrina nas seguintes concentrações (mg i.a./ml): 10ù7, 10ù6, 10ù5, 10ù4, 10ù3. O comprimento dos ovários e o número de ovócitos foram quantificados após a primeira postura. O comprimento do ovário variou de 5,7 mm na concentração de 10ù4 mg i.a./ml a 6,4 mm na concentração de 10ù7 mg i.a./ml, com valores semelhantes para as outras concentrações de permetrina e controle. O número de ovócitos por fêmea variou de 13,5 (10ù3 mg i.a./ml) a 29,2 no controle, com diferenças significativas entre concentrações. O número de ovócitos por fêmea, proveniente de ninfas expostas a concentração de 10ù5 mg i.a./ml, foi semelhante àquele do controle. No entanto, o menor número de ovócitos por fêmea, provenientes de ninfas expostas a outras concentrações de permetrina, sugere que esse inseticida pode afetar a capacidade reprodutiva. Esses resultados são discutidos em relação à tolerância de Heteroptera predador aos inseticidas e a possível ocorrência de hormese.

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.

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.

Controle da cigarrinha-verde (Empoasca kraemeri (Ross & Moore, 1957) (Hemiptera - Homoptera: cicadellidae)) no feijoeiro (Phaseolus vulgaris) / Control of cigarrinha-verde (Empoasca kraemeri (Ross & Moore, 1957) (Hemiptera - Homoptera: cicadellidae)) in the beans (Phaseolus vulgaris)

Estudou-se os efeitos tóxicos dos inseticidas esfenvalerato, fenitrotion, fenpropatrin e monocrotofos sobre as ninfas de E. kraemeri em cultura de feijoeiro. A pesquisa doi conduzida no município de Nepomuceno - MG, utilizando uma lavoura comerical de cultivar "Carioca". Os compostos foram diluídos em água nas doses recomendadas pelos fabricantes para o controle de pragas, sendo a aplicaçäo realizada através de um pulverizador costal, propiciando volume de aplicaçäo de 400 L/ha. Os resultados mostraram que todos os compostos controlaram a praga em questäo, porém o fenitrotion na dose de 750 g i.a./ha apresentou o maior nível médio de controle.

Changes in the integument of nymphal and adult hyalomma [hyalomma] dromedarii koch[Acari: Ixodoidea: Ixodidae] II the effect of 20-hydroxy ecdysone

The exogenous doze of 20-Hydroxy Ecdysone [20-HE] used in the present study did not accelerate moulting in nymphal Hyalomma dromedarii. However, histological observations exhibit an increase in epidermal cell activity and in the deposition of the endocuticular lamellae

effect of 20-hydoxyecdysone on the neurohemal endocrine organs in Hyalomma Domedarii Koch [ixodoidea: ixodidae]

Twenty ug of 20-hydroxyecdysone [20-HE] were applied topically to nymphal Hyalomma dromedarii on the day of detachment. The effect of 20-hydroxyecdysone was studied on the lateral segmental organs and on the neurohemal organs in the emerging unfed, semifed and fully engorged females, and compared with those in the untreated females. The effect induced by 20-hydroxyecdysone included [a] The lateral segmental organs in each group of treated female were comparatively larger than that in the untreated ones, and [b] the dorsal lobes of the neurohemal organs were comparatively smaller. The results suggest that the lateral segmental organs play a role in ecdysone production