RNA interference of the clock gene period disrupts circadian rhythms in the expression of genes related to insecticide resistance in the chagas disease vector Triatoma infestans (Hemiptera: Reduviidae).

In Triatoma infestans it was observed pyrethroid resistance attributed in part to an elevated oxidative metabolism mediated by cytochromes P450. The nicotinamide adenine dinucleotide phosphate (NADPH) cytochrome P450 reductase (CPR) plays a crucial role in catalysing the electron transfer from NADPH to all cytochrome P450s. The daily variations in the expression of CPR gene and a P450 gene (CYP4EM7), both associated with insecticide resistance, suggested that their expressions would be under the endogenous clock control. To clarify the involvement of the clock in orchestration of the daily fluctuations in CPR and CYP4M7 genes expression, it was proposed to investigate the effect of silencing the clock gene period (per) by RNA interference (RNAi). The results obtained allowed to establish that the silencing of per gene was influenced by intake schemes used in the interference protocols. The silencing of per gene in T. infestans reduced its expression at all the time points analysed and abolished the characteristic rhythm in the transcriptional expression of per mRNA. The effect of the per gene silencing in the expression profiles at the transcriptional level of CPR and CYP4EM7 genes showed the loss of rhythmicity and demonstrated the biological clock involvement in the regulation of t heir expression.

Modelling the association between deltamethrin resistance in Triatoma infestans populations of the Argentinian Gran Chaco region with environmental factors.

In Latin America, Triatoma infestans is the main vector of the protozoan Trypanosoma cruzi, causal agent of Chagas disease. This blood-sucking triatomine is widely distributed in the Gran Chaco ecoregion, where chemical control has failed because of the evolution of resistance to pyrethroid insecticides. Recently, we described a deltamethrin high resistant focus in Güemes Department (Chaco province) characterized by susceptible populations, populations with low resistance (without field control failures) and some of the populations with the highest resistance level detected. This toxicological heterogeneity could be a result of non-homogenous insecticide pressure and be influenced by environmental factors. The present study evaluated the association of deltamethrin resistance ratios (RR50s) of T. infestans populations with explanatory variables extracted from the WorldClim dataset and constructed from information of National Chagas Program actions during 2005-2015. Control actions were distributed throughout the analyzed period, representing a homogeneous selective pressure. The average percentage of total positive houses was 33.66%. Models that included temperature and precipitation indicators presented 65% explanation. When village size variables where added, the explanatory power reached 70%. This observational result suggests that the climate may favor directly or indirectly the development/selection for resistance, representing a valuable tool to understand the occurrence of resistance that could increase the Chagas disease in the Gran Chaco.

Geographical Variation of Deltamethrin Susceptibility of Triatoma infestans (Hemiptera: Reduviidae) in Argentina With Emphasis on a Resistant Focus in the Gran Chaco.

Chagas disease is one of the most important parasitic infections in Latin America. The main vector of the protozoan Trypanosoma cruzi in America is Triatoma infestans, a blood-sucking triatomine bug who is widely distributed in the Gran Chaco ecoregion. Control programs in endemic countries are focused in the elimination of triatomine vectors with pyrethroid insecticides. However, chemical control has failed in the Gran Chaco over the last two decades because of several factors. Previous studies have reported the evolution of different levels of resistance to deltamethrin in Tri. infestans Recently, very high resistance has been found in the central area of the Argentine Gran Chaco. However, the origin and the extension of this remarkably resistant focus remain unknown. The aim of this study was to evaluate the geographical variation of deltamethrin susceptibility of Tri. infestans in different endemic provinces of Argentina, with emphasis in the center of the Argentine Gran Chaco ecoregion where this main vector has not been reduced. Populations of Mendoza, San Juan, Santiago del Estero, and Tucumán provinces were all susceptible. Resistant populations were only detected in the province of Chaco, where a mosaic resistant focus was described at the Güemes Department. It was characterized into three pyrethroid resistance categories: susceptible, low, and highly resistant populations. We found the populations with the highest resistance levels to deltamethrin, with resistant ratios over 1000.

Effect of Environmental Conditions and Toxic Compounds on the Locomotor Activity of Pediculus humanus capitis (Phthiraptera: Pediculidae).

In this work, we evaluated the effect of environmental variables such as temperature, humidity, and light on the locomotor activity of Pediculus humanus capitis. In addition, we used selected conditions of temperature, humidity, and light to study the effects of cypermethrin and N,N-diethyl-3-methylbenzamide (DEET) on the locomotor activity of head lice. Head lice increased their locomotor activity in an arena at 30°C compared with activity at 20°C. When we tested the influence of the humidity level, the locomotor activity of head lice showed no significant differences related to humidity level, both at 30°C and 20°C. Concerning light influence, we observed that the higher the intensity of light, the slower the movement of head lice. We also demonstrated that sublethal doses of toxics may alter locomotor activity in adults of head lice. Sublethal doses of cypermethrin induced hyperactivated responses in adult head lice. Sublethal doses of DEET evocated hypoactivated responses in head lice. The observation of stereotyped behavior in head lice elicited by toxic compounds proved that measuring locomotor activity in an experimental set-up where environmental conditions are controlled would be appropriate to evaluate compounds of biological importance, such as molecules involved in the host-parasite interaction and intraspecific relationships.

New findings of insecticide resistance in Triatoma infestans (Heteroptera: Reduviidae) from the Gran Chaco.

Chagas disease is a major health concern in Latin America, and Triatoma infestans (Klug, 1834) is responsible for the majority of cases of Chagas disease in the continent. After the discovery of deltamethrin highly resistant populations in the neighboring areas to Yacuiba (Bolivia) and Salvador Mazza (Argentina), we studied T. infestans populations north and southward, with the aim of describing the range of the resistant area. In addition, tests were conducted to describe the susceptibility to fipronil in deltamethrin-resistant populations. Tarija populations were highly resistant to deltamethrin, showing that the resistant area is greater than previously reported. Argentinean and Paraguayan populations were susceptible or presented moderate to low levels of deltamethrin resistance. Resistance to fipronil was found in Bolivian populations, although this insecticide was effective against Argentinean populations. This study shows that the Argentinean and Paraguayan populations are currently under successful control with deltamethrin. However, continuous resistance monitoring is necessary. We found that fipronil is a viable option for Argentina, whereas the resistance found in Bolivia implies that new formulations are needed to control T. infestans in Bolivia. Further research is required to find new alternatives of control in those areas that are currently suffering from high infestation rates.

Resistance to insecticides and effect of synergists on permethrin toxicity in Pediculus capitis (Anoplura: Pediculidae) from Buenos Aires.

Permethrin-resistant colonies of Pediculus capitis (De Geer) from Buenos Aires were used to establish a resistance profile and to examine resistance mechanisms. All permethrin-resistant head lice (resistance ratio from 52.8 to > 88.7) were also resistant to d-phenothrin (resistance ratio from 40.86 to > 48.39) and deltamethrin (resistance ratio from 16.24 to 38.06). No cross-resistance to carbaryl was found in any of the pyrethroid-resistant P. capitis tested. Otherwise, all resistant colonies showed low to high levels of resistance to beta-cypermethrin. This pyrethroid had never been applied as a pediculicide in Argentina; however, the high level of resistance found in these permethrin-resistant colonies (resistance ratio from 9.74 to 50.97) demonstrated that pyrethroid cross-resistance occurred to this novel insecticide. Treatment with piperonyl butoxide (PBO) or triphenylphosphate (TPP) significantly decreased the toxicity of permethrin in the four colonies tested. The esterase inhibitor TPP produced lower enhancement of toxicity than the multifunction oxidase inhibitor PBO in the colonies having the highest resistance levels. Results presented here concerning the cross-resistance profile and synergism by enzyme inhibitors in permethrin-resistant head lice demonstrated that enhanced metabolism was involved in the pyrethroid resistance. However, the substantial degree of resistance that remained after synergism suggested the presence of another resistance mechanism. Cross-resistance to pyrethroid and susceptibility to the carbamate carbaryl suggested a common action mechanism.