Cuticular extracts induce aggregation in head lice.

The human head lice Pediculus humanus capitis (De Geer) (Phthiraptera: Pediculidae) are strict, obligate human ectoparasites that spends their entire life cycle in the host and cause skin irritation and derived infections. Despite the health-related importance, few studies have evaluated the chemical communication among these insects. Here, we evaluate the response of lice of both sexes to cuticular extracts using two solvents of different polarity (hexane and methanol). Cuticular extracts that elicited an attraction response towards head lice were analysed by gas chromatography-mass spectrometry (GC-MS) to determine the cuticular lipid profile. Both lice sexes were attracted to the hexane extracts but not the methanol extracts, suggesting the non-polarity of the compounds present in the cuticle. Chemical analyses of hexane extracts from males and females showed high similarity in major compounds. This study provides the first evidence that lice respond to cuticle extracts, which may be important to understand aggregation behaviour.

Head lice were also affected by COVID-19: a decrease on Pediculosis infestation during lockdown in Buenos Aires.

Pediculosis is a worldwide disease affecting school-aged children produced by the presence of the head louse, Pediculus humanus capitis De Geer, an obligate ectoparasite on the human scalp feeding exclusively on blood. Transmission occurs primarily through direct physical head-to-head contact. In March 2020, the World Health Organization (WHO) declared the COVID-19 outbreak as a pandemic. COVID-19 is caused by SARS-CoV-2, a variant of the coronavirus. Therefore, on March 18, 2020, the Argentinean government established mandatory isolation for an indefinite period. This obligatory isolation interrupted regular classes avoiding direct contact between children, thus affecting the dispersal route of individuals and the evolution of head louse populations. In this study, we evaluated through an online survey how confinement affected the prevalence of lice during lockdown compared to the situation prior to confinement. The survey allowed to discriminate the different control strategies, the number of treatments, and the amount of insects recorded by parents. Data of 1118 children obtained from 627 surveys were analyzed. As the main result, it was observed that prevalence of lice decreased significantly from before (69.6%) to during (43.9%) COVID-19 lockdown. Moreover, head lice infestation was more effectively controlled in households with up to 2 children in comparison to households with 3 or more children. This is the first study that analyzed the prevalence of head lice during COVID-19 pandemic. In addition, this work demonstrated the impact of social distance in the population dynamics of head lice and how it could affect the control strategies in the future.

Mouthpart sensory structures of the human head louse Pediculus humanus capitis.

Head lice are exclusive human parasitic blood-sucking insects. Distributed worldwide among school-age children, this parasitosis generates scalp irritation and sometimes social prejudice. Understanding how head lice detect and perceive their human hosts is crucial to control transmission. Here, we describe the sensory structures present on the mouthparts of Pediculus humanus capitis and their possible contribution to the feeding decision-making process. On the anterior zone of the clypeus around the haustellum two morphological types of sensilla, invariable in location and number, were identified: fourteen short clypeus bristles (SCB) and six long clypeus bristles (LCB). During feeding these structures contact the host skin but not its blood. Located antero-dorsally on the everted haustellum and between the epipharyngeal teeth, a third sensillar type was identified: about four short peg epipharyngeal (SPE) sensilla. These structures are bathed with the incoming blood, when head lice feed, so may have a gustatory role. In behavioural experiments antennectomy of lice did not interfere with feeding behaviour, suggesting that the sensory structures on the mouthparts could be involved in host assessment.

The Sensory Machinery of the Head Louse Pediculus humanus capitis: From the Antennae to the Brain.

Insect antennae are sophisticated sensory organs, usually covered with sensory structures responsible for the detection of relevant signals of different modalities coming from the environment. Despite the relevance of the head louse Pediculus humanus capitis as a human parasite, the role of its antennal sensory system in the highly dependent relation established with their hosts has been barely studied. In this work, we present a functional description of the antennae of these hematophagous insects by applying different approaches, including scanning electron microscopy (SEM), anterograde antennal fluorescent backfills, and behavioral experiments with intact or differentially antennectomized lice. Results constitute a first approach to identify and describe the head louse antennal sensilla and to determine the role of the antenna in host recognition. SEM images allowed us to identify a total of 35-40 sensilla belonging to seven different morphological types that according to their external architecture are candidates to bear mechano-, thermo-, hygro-, or chemo-receptor functions. The anterograde backfills revealed a direct neural pathway to the ipsilateral antennal lobe, which includes 8-10 glomerular-like diffuse structures. In the two-choice behavioral experiments, intact lice chose scalp chemicals and warm surfaces (i.e., 32°C) and avoided wet substrates. Behavioral preferences disappeared after ablation of the different flagellomeres of their antenna, allowing us to discuss about the location and function of the different identified sensilla. This is the first study that integrates morphological and behavioral aspects of the sensory machinery of head lice involved in host perception.

Ivermectin lipid-based nanocarriers as novel formulations against head lice.

The use of pyrethroids to control the human head louse, Pediculus humanus capitis De Geer (Anoplura: Pediculidae), has suffered considerable loss of efficacy due to the evolution of resistance. Thus, the development of efficiently insecticide delivery systems is imperative for the control of head lice. We studied the insecticidal activity of ivermectin-loaded lipid nanocapsules (IVM-LNC) against permethrin-resistant head lice from Argentina. The LNC, prepared by a phase inversion procedure, were characterized in terms of size, surface potential, and physical stability. These nanoparticles were nearly spherical with mean diameters of 55 nm and narrow size distribution (PI ≤ 0.2). The KT50 mortality values of head lice after exposure to two IVM-LNC formulations (0.11 and 0.28%) were significantly smaller (5 and 3 h, respectively) compared to those exposed only to LNC control group (8 h). This investigation showed the effectiveness in the encapsulation of ivermectin (IVM) into stable LNC dispersion with a potential clinical activity against head lice.

Identifying Current and Missing Knowledge in the Control of Pyrethroid-Resistant Triatoma infestans, Vector of Chagas Disease

Triatomines are blood-sucking bugs that occur mainly in Latin America. They are vectors of Trypanosoma cruzi, the parasite that causes Chagas disease. Chemical control of Chagas disease´s vectors by using pyrethroid insecticides has been highly successful for the elimination of domestic infestation and consequently the reduction of the vector transmission. However, at the beginning of the 2000s a decrease in the effectiveness of the chemical control of triatomines was detected in several areas from Argentina and Bolivia, particularly in the Gran Chaco eco-region. During the last 15 years, several studies demonstrated the evolution of insecticide resistance in Triatoma infestans and established the presence of different toxicological profiles, the autosomal inherence of resistance, the biological costs of deltamethrin resistance, the expression of deltamethrin resistance thorough the embryonic development, and the main mechanisms of resistance (target-site insensitivity and metabolic detoxification of insecticides). The emergence of pyrethroid resistance coupled with the usual difficulties in sustaining adequate rates of insecticide applications emphasize the need of incorporating other tools for integrated vector and disease control, such as the proposal of the organo-phosphorus insecticide fenitrothion as an alternative chemical strategy for the management of the resistance because it was effective against pyrethroid-resistant populations in laboratory and semi-field trials. New studies on the current situation of presence and spread of resistant populations of triatomines and the acceptance of the use of alternative insecticides are critical requirements in the implementation of strategies for the management of resistance and for the rational design of campaigns oriented to reducing the vector transmission of Chagas' disease.

Demographic effects of deltamethrin resistance in the Chagas disease vector Triatoma infestans

Triatoma infestans (Heteroptera: Reduviidae) Klug is the main vector of Chagas disease in Latin America. Resistance to deltamethrin was reported in Argentina and recently associated with reproductive and longevity trade-offs. The objectives of the present study were to describe the demographic consequences of deltamethrin resistance in T. infestans and to establish possible target stages for chemical control in susceptible and resistant colonies. A stage-classified matrix model was constructed based on the average stage length for susceptible, resistant and reciprocal matings' progeny. The differences between colonies were analysed by prospective and retrospective analysis. The life table parameters indicated reduced fecundity, fertility and population growth in resistant insects. The retrospective analysis suggested the latter was associated with lower reproductive output and increased fifth-instar nymph stage length. The prospective analysis suggested that the adult stage should be the main target for insecticide control. Although, fifth-instar nymphs should also be targeted when resistance has been detected. The presented results show demographic effects of deltamethrin resistance in T. infestans. While the older stages could be the main targets for chemical control, this approach is impeded by their higher tolerance to insecticides. It is concluded that the different mode of action insecticides would be more effective than a dose increase for the control of deltamethrin-resistant T. infestans.

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.

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.

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.

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.

Influence of the formulations in removing eggs of Pediculus humanus capitis (Phthiraptera: Pediculidae).

Head lice lay eggs in human head hairs in order to reproduce. There is a difficulty associated to the process of detaching these eggs: they are tightly gripped to the hair by a secretion produced by female head lice. The physical removal of eggs has become an important part of treatment of louse infestations. The finding of new products to loosen the eggs is necessary to avoid mistaken diagnosis or reinfestations. This work aimed to compare different kinds of pediculicide formulations in order to find if their presentations represented differences in the egg remover effect. We also wanted to present a new device to test the efficacy of the egg remover formulations. Products with creamy presentations (Bio infant lice and egg remover and hair conditioner) and one containing dimethicone (Nyda) showed the lower mean forces compared with the control (lower mean forces represented best removal activity). Whereas, the Biferdil egg remover (gel) and Nopucid Tribit (hydroalcoholic lotion) had no egg removal effect, presenting the highest mean forces (177.82 and 189.99 mN, respectively) compared with the control. Additionally, we proposed a removal index (RI) to compare the efficacy of different products on the egg removal activity (RI > 0, good performance). The higher index values were for Bio infant lice and egg remover (0.72) and Biferdil hair conditioner (0.58). The lowest index values were for Biferdil egg remover (-0.26) and Nopucid Tribit (-0.35).The formulation of over the counter pediculicides in the egg remover effect was discussed.

Geographical distribution of pyrethroid resistance allele frequency in head lice (Phthiraptera: Pediculidae) from Argentina.

The human head louse, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae), is an obligate ectoparasite that causes pediculosis capitis and has parasitized humans since the beginning of humankind. Head louse infestations are widespread throughout the world and have been increasing since the early 1990s partially because of ineffective pediculicides. In Argentina, the overuse of products containing pyrethroids has led to the development of resistant louse populations. Pyrethroid insecticides act on the nervous system affecting voltage-sensitive sodium channels. Three point mutations at the corresponding amino acid sequence positions M815I, T917I, and L920F in the voltage-gated sodium channel gene are responsible for contributing to knockdown resistance (kdr). The management of pyrethroid resistance requires either early detection or the characterization of the mechanisms involved in head louse populations. In the current study, we estimated the distribution of kdr alleles in 154 head lice from six geographical regions of Argentina. Pyrethroid resistance kdr alleles were found in high frequencies ranging from 67 to 100%. Of these, 131 (85.1%) were homozygous resistant, 13 (8.4%) were homozygous susceptible, and 10 (6.5%) were heterozygous. Exact tests for the Hardy-Weinberg equilibrium for each location showed that genotype frequencies differed significantly from expectation in four of the six sites studied. These results show that pyrethroid resistance is well established reaching an overall frequency of 88%, thus close to fixation. With 30 yr of pyrethroid-based pediculicides use in Argentina, kdr resistance has evolved rapidly among these head louse populations.

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.

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.

Expression of insecticide resistance in immature life stages of Triatoma infestans (Hemiptera: Reduviidae).

The aim of the current study was to investigate the susceptibility to the insecticide deltamethrin and the expression of resistance to this insecticide in developing eggs and neonate nymphs of Triatoma infestans from two areas of Argentina (Campo Largo) and Bolivia (Entre Ríos), where resistance to this pyrethroid is suspected. Both nymphal populations showed resistance to deltamethrin, with lower resistance ratio for Entre Ríos (173X) than Campo Largo (1108X). Efficacy of deltamethrin on 4-, 7-, and 12-d-old eggs for both field populations were significantly lower than efficacy on eggs of the susceptible strain. This is the first documented evidence of the expression of pyrethroid resistance during the embryonic development of Chagas Disease vectors.

Microgeographical study of insecticide resistance in Triatoma infestans from Argentina.

Chagas disease is a chronic parasitic infection restricted to America where it is currently estimated that 90 million people are at risk of acquiring the infection. Chemical control with pyrethroid insecticides has been effective to reduce disease transmission in several areas of the Southern Cone, although insecticide resistance has evolved and diminished the campaigns' results. Considering previous reports on the different levels of resistance between Triatoma infestans from different geographical areas, the objective of this work was to determine if T. infestans populations are toxicologically structured within localities. Response to the insecticide was measured and compared between houses of two Argentine localities. Different toxicity of deltamethrin was detected between dwellings of Chaco province, accounting for both susceptible and resistant houses within the same locality. However no difference was found among houses of Salta province. The results obtained in this work suggest that geographical structure is present not only at the between localities level, but also at the microgeograhical 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).

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 13 mm in KSL® plastic, 0.3 and 10.7 mm in NOPUCID® plastic, 0.15 and 31 mm in KSL® metal and 0.09 and 37 mm 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.