Gastón Mougabure-Cueto

Insecticide resistance in vector Chagas disease: Evolution, mechanisms and management

Chagas disease is a chronic parasitic infection restricted to America. The disease is caused by the protozoa Trypanosoma cruzi, which is transmitted to human through the feces of infected triatomine insects. Because no treatment is available for the chronic forms of the disease, vector chemical control represents the best way to reduce the incidence of the disease. Chemical control has been based principally on spraying dwellings with insecticide formulations and led to the reduction of triatomine distribution and consequent interruption of disease transmission in several areas from endemic region. However, in the last decade it has been repeatedly reported the presence triatomnes, mainly Triatoma infestans, after spraying with pyrethroid insecticides, which was associated to evolution to insecticide resistance. In this paper the evolution of insecticide resistance in triatomines is reviewed. The insecticide resistance was detected in 1970s in Rhodnius prolixus and 1990s in R. prolixus and T. infestans, but not until the 2000s resistance to pyrthroids in T. infestans associated to control failures was described in Argentina and Bolivia. The main resistance mechanisms (i.e. enhanced metabolism, altered site of action and reduced penetration) were described in the T. infestans resistant to pyrethrods. Different resistant profiles were demonstrated suggesting independent origin of the different resistant foci of Argentina and Bolivia. The deltamethrin resistance in T. infestans was showed to be controlled by semi-dominant, autosomally inherited factors. Reproductive and developmental costs were also demonstrated for the resistant T. infestans. A discussion about resistance and tolerance concepts and the persistence of T. infestans in Gran Chaco region are presented. In addition, theoretical concepts related to toxicological, evolutionary and ecological aspects of insecticide resistance are discussed in order to understand the particular scenario of pyrethroid resistance in triatomines.

Identification of a point mutation associated with pyrethroid resistance in the para-type sodium channel of Triatoma infestans, a vector of Chagas' disease.

The voltage-gated sodium channel is the target site of pyrethroid insecticides. Point mutations in the domain II region of the channel have been implicated in pyrethroid resistance of several insect species. We identified the sequence of domain II from the para sodium channel in Rhodnius prolixus, a vector of Chagas disease. With this information, we cloned and sequenced the domain II of the sodium channel from the other main Chagas disease vector: Triatoma infestans. We also identified the presence of a resistance-conferring mutation (L1014F) in a pyrethroid-resistant population of T. infestans from Argentina, and present a PCR-based method to detect this mutation in individuals from field populations. These findings have important implications for the implementation of strategies for resistance management, and for the rational design of campaigns for the control of Chagas disease transmission.

Toxicity of non-pyrethroid insecticides against Triatoma infestans (Hemiptera: Reduviidae)

Triatoma infestans (Klug) is the main vector of Chagas disease, which is a public health concern in most Latin American countries. The prevention of Chagas disease is based on the chemical control of the vector using pyrethroid insecticides. In the last decade, different levels of deltamethrin resistance have been detected in certain areas of Argentina and Bolivia. Because of this, alternative non-pyrethroid insecticides from different chemical groups were evaluated against two T. infestans populations, NFS and El Malá, with the objective of finding new insecticides to control resistant insect populations. Toxicity to different insecticides was evaluated in a deltamethrin-susceptible and a deltamethrin-resistant population. Topical application of the insecticides fenitrothion and imidacloprid to first nymphs had lethal effects on both populations, producing 50% lethal dose (LD50) values that ranged from 5.2-28 ng/insect. However, amitraz, flubendiamide, ivermectin, indoxacarb and spinosad showed no insecticidal activity in first instars at the applied doses (LD50 > 200 ng/insect). Fenitrothion and imidacloprid were effective against both deltamethrin-susceptible and deltamethrin-resistant populations of T. infestans. Therefore, they may be considered alternative non-pyrethroid insecticides for the control of Chagas disease.

L925I mutation in the Para-type sodium channel is associated with pyrethroid resistance in Triatoma infestans from the Gran Chaco region

Background Chagas disease is an important public health concern in Latin America. Despite intensive vector control efforts using pyrethroid insecticides, the elimination of Triatoma infestans has failed in the Gran Chaco, an ecoregion that extends over Argentina, Paraguay, Bolivia and Brazil. The voltage-gated sodium channel is the target site of pyrethroid insecticides. Point mutations in domain II region of the channel have been implicated in pyrethroid resistance of several insect species. Methods and Findings In the present paper, we identify L925I, a new pyrethroid resistance-conferring mutation in T. infestans. This mutation has been found only in hemipterans. In T. infestans, L925I mutation occurs in a resistant population from the Gran Chaco region and is associated with inefficiency in the control campaigns. We also describe a method to detect L925I mutation in individuals from the field. Conclusions and Significance The findings have important implications in the implementation of strategies for resistance management and in the rational design of campaigns for the control of Chagas disease transmission.

Fenitrothion: an alternative insecticide for the control of deltamethrin-resistant populations of Triatoma infestans in northern Argentina.

Deltamethrin‐based campaigns to control Triatoma infestans (Klug) (Hemiptera: Reduviidae) have decreased in success as a result of the development of insecticide resistance. We compared the in vitro effects of the pyrethroid deltamethrin and two doses of the organophosphate fenitrothion, presented on different materials, on T.u2009infestans from La Esperanza, Argentina. Laboratory tests demonstrated a decrease in susceptibility to deltamethrin in the field population [LD50: 30.32u2009nanograms per insect (ng/i)] compared with the reference population (LD50: 0.13u2009ng/i), giving a high resistance ratio of 233.42. By contrast, similar susceptibility to fenitrothion was assessed in both the field and reference populations (LD50: 21.65u2009ng/i and 21.38u2009ng/i, respectively). The effectiveness of the formulated insecticides varied according to the surfaces to which they were applied. The application of fenitrothion formulations to glass or brick resulted in mortality of 90–100%. The application of fenitrothion formulations to wood or mud caused mortality in the range of 6.7–56.7%. Resistant insects presented low mortality when exposed to the deltamethrin formulation and high mortality when exposed to fenitrothion formulations. Moreover, the insecticides demonstrated residual activity only when applied to glass. The present work demonstrates that fenitrothion is an alternative to pyrethroids for the management of deltamethrin‐resistant insects in La Esperanza. However, this effectiveness is not sustained over time.

The fumigant and repellent activity of aliphatic lactones against Pediculus humanus capitis (Anoplura: Pediculidae)

New alternative insecticides are necessary for the chemical control of head lice. In this study the fumigant knockdown time 50% (KT50) and repellency index (RI) of three aliphatic lactones was compared with two essential oils and DDVP, against permethrin-resistance Pediculus humanus capitis from Argentina. In the fumigant assay, none of the lactones were effective compared to the highest activity of eucalyptus (KT50 15.53 m). In the repellency test, the three lactones were equally or more effective (RI ranging from 60.50 to 76.68) than the positive control (piperonal). These lactones are promising as head lice repellents.

Comparative efficacy of commercial combs in removing head lice (Pediculus humanus capitis) (Phthiraptera: Pediculidae)

The use of a fine comb for removing lice from the head of the human host is a relevant tool both in the diagnosis of infestations and as part of an integrated control strategy of head lice. The effectiveness of a fine comb depends, in part, on the design and material they are built. The aim of this study was to compare in vivo the efficacy of metal and plastic combs that are currently used in the removal of head lice and eggs worldwide. The space between comb teeth and the length was 0.23 and 13xa0mm in KSL® plastic, 0.3 and 10.7xa0mm in NOPUCID® plastic, 0.15 and 31xa0mm in KSL® metal and 0.09 and 37xa0mm in ASSY® metal. The assays were performed comparing the combs in pairs: (a) KSL® vs. NOPUCID® plastic combs, (b) KSL® vs. ASSY® metal combs and (c) KSL® plastic comb vs. ASSY® metal comb. The most effective plastic comb was KSL®, removing a higher number of individuals of all stages. The most effective metal comb was ASSY®, removing more insects of all stages (except adults). The comparative test between KSL® plastic and ASSY® metal showed that ASSY® was the most effective in removing head lice and their eggs.

Comparative toxicity of oxygenated monoterpenoids in experimental hydroalcoholic lotions to permethrin-resistant adult head lice

The use of botanical compounds such as essential oils has recently become the subject of great interest as a natural means of pest control because of their ovicidal, larvicidal, or adulticidal activity against various insect species including head lice. We tested and compared the efficacy of pure oxygenated monoterpenoids that are main ingredients of essential oils of good biological activity. We used pulegone and citral, components of Aloysia citrodora, and geraniol, citronellol, and linalool, components of Geranium sp. oil. We found that citronellol and geraniol showed the highest knockdown and mortality effect (>60%) on adults of both sexes (50:50%) and third-stage nymphs. Pulegone, linalool, and citral showed knockdown percentages between 42 and 55%, and mortality percentages between 47 and 53%. A simple linear regression analysis showed statistically significant relationships between the studied toxic effects and viscosity of the monoterpenoids (pxa0<xa00.05), but not with their partition coefficient (log P).

Identification of three cytochrome P450 genes in the Chagas' disease vector Triatoma infestans: Expression analysis in deltamethrin susceptible and resistant populations

Cytochrome P450 monooxygenases play a predominant role in the metabolism of insecticides. Many insect P450 genes have frequently been associated with detoxification processes allowing the insect to become tolerant or resistant to insecticides. The increases of expression of P450 genes at transcriptional level are often consider responsible for increasing the metabolism of insecticides and seems to be a common phenomenon in the evolution of resistance development in insects. As pyrethroid resistance has been detected in Triatoma infestans, it was of interest to analyze genes associated with resistance to insecticides such as those encoding for cytochromes P450. With this purpose, the cDNA sequences of three cytochrome P450 genes (CYP4EM7, CYP3085B1, and CYP3092A6) were identified in this species. Primers and specific Taqman probes were designed from these sequences to determine their expression by quantitative PCR. The mRNA levels of the cytochrome P450 genes identified were determined from total RNA extracted from pools of fat body collected from individuals of different resistant and susceptible strains of T. infestans, and at different interval times after the topical application of the lethal doses 50% (LD50) of deltamethrin on the ventral abdomen of insects belonging to the different populations analyzed. It was detected overexpression of the CYP4EM7 gene in the most resistant strain of T. infestans and the expression of the three cytochrome P450 genes isolated was induced by deltamethrin in the susceptible and resistant populations included in this study. These results suggest that these genes would be involved in the detoxification of deltamethrin and support the hypothesis that considers to the cytochrome P450 genes of importance in the development of pyrethroid resistance.

Lethal activity of individual and mixed monoterpenoids of geranium essential Oil on Musca domestica

Plant essential oils and its constituent molecules have been suggested as an alternative to control insect. The contribution of the constituents to the effect of the oil is determined by the interactions occurring between them. Synergistic interactions would improve the insecticide efficacy of the compounds due to the utilization of lower doses. We evaluated the insecticidal activity of geranium (Geranium maculatum L.) oil and its major constituents against Musca domestica L. and studied the toxic interactions in artificial mixtures of those constituents in the natural ratio. While synergistic interactions were determined in house fly in this study, these were of low intensity evidencing that the effect of each constituent was slightly modified by the other constituents present in the mixtures. The search for synergism between components is a strategy to improve the insecticide activity of natural compounds. The synergism helps to reduce the environmental and toxicological impact due to the reduction of the dose of use.