Locomotor activity in Aedes aegypti with different insecticide resistance profiles

ABSTRACT OBJECTIVE: To evaluate locomotor activity in four field populations of Ae. aegypti with different insecticide resistance profiles from the state of São Paulo for two years. METHODS: This study comprised the susceptible Rockefeller strain and four populations from São Paulo, Brazil: two considered populations with "reduced susceptibility" to pyrethroids (Campinas and Marília), and two "resistant populations" (Santos and Ribeirão Preto). First, 2016 and 2017 eggs from these five populations were hatched in laboratory. Virgin females underwent experiments under laboratory conditions at 25°C, with 12:12h light/dark (LD) photoperiod; 24-hour individual activity was recorded using a locomotor activity monitor (LAM). RESULTS: In females from 2016 field populations, both resistant populations showed significant more locomotor activity than the two reduced susceptibility populations and the Rockefeller strain (p < 0.05). As for females from 2017 field populations, reduced susceptibility populations showed a significant increased locomotor activity than the Rockefeller strain, but no significant difference when compared to Santos resistant population (p > 0.05). CONCLUSIONS: Our results indicate that insecticide-resistant Ae. aegypti populations show increased locomotor activity, which may affect the transmission dynamics of their arboviruses.

Tolerance of dark septate endophytic fungi (DSE) to 1 agrochemicals in vitro / Tolerancia de hongos endofíticos septados oscuros a agroquímicos in vitro

Abstract Dark septate endophytes (DSE) are a heterogeneous group of fungi, mostly belonging to the Phylum Ascomycota, that are involved in a mutualistic symbiosis with plant roots. The aim of this study is to evaluate the behavior of two strains of DSE isolated from wheat roots of two cropping areas in the province of Buenos Aires, Argentina, against some agrochemicals. Of all the isolates obtained, two strains were identified as Alternaria alternata and Cochliobolus sp. These DSE were found to be tolerant to glyphosate, carbendazim and cypermethrin when evaluated at the recommended agronomic dose (AD), 2 AD and, in some cases, 10 AD. This work contributes to the study of the biology of this group of fungi and their tolerance in the presence of xenobiotics widely used in agriculture.© 2019 Asociaci´on Argentina de Microbiolog´ıa. Published by Elsevier Espa˜na, S.L.U. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/bync-nd/4.0/).

Resumen Los endófitos septados oscuros (DSE) son un grupo heterogéneo de hongos que participan de una simbiosis mutualista con raíces de plantas, perteneciendo principalmente al Phylum Ascomycota. El objetivo de este estudio fue aislar DSE de raíces de trigo proveniente de dos áreas de cultivo de la provincia de Buenos Aires y evaluar el comportamiento de dos cepas de DSE aisladas de raíces de trigo frente a algunos agroquímicos en dos áreas de cultivo de la provincia de Buenos Aires. De todos los aislamientos obtenidos se seleccionaron dos cepas que se identificaron como Alternaria alternata y Cochliobolus sp. Se encontró que estos DSE son tolerantes al glifosato, el carbendazim y la cipermetrina, evaluados a las dosis agronómicas recomendadas (AD), a 2x AD y, en algunos casos, a 10x AD. Este trabajo contribuye al conocimiento de la biología de este grupo de hongos y su tolerancia a xenobióticos ampliamente utilizados en la agricultura.

Spatiotemporal multiple insecticide resistance in Aedes aegypti populations in French Guiana: need for alternative vector control

BACKGROUND Aedes aegypti is the sole vector of urban arboviruses in French Guiana. Overtime, the species has been responsible for the transmission of viruses during yellow fever, dengue, chikungunya and Zika outbreaks. Decades of vector control have produced resistant populations to deltamethrin, the sole molecule available to control adult mosquitoes in this French Territory. OBJECTIVES Our surveillance aimed to provide public health authorities with data on insecticide resistance in Ae. aegypti populations and other species of interest in French Guiana. Monitoring resistance to the insecticide used for vector control and to other molecule is a key component to develop an insecticide resistance management plan. METHODS In 2009, we started to monitor resistance phenotypes to deltamethrin and target-site mechanisms in Ae. aegypti populations across the territory using the WHO impregnated paper test and allelic discrimination assay. FINDINGS Eight years surveillance revealed well-installed resistance and the dramatic increase of alleles on the sodium voltage-gated gene, known to confer resistance to pyrethroids (PY). In addition, we observed that populations were resistant to malathion (organophosphorous, OP) and alpha-cypermethrin (PY). Some resistance was also detected to molecules from the carbamate family. Finally, those populations somehow recovered susceptibility against fenitrothion (OP). In addition, other species distributed in urban areas revealed to be also resistant to pyrethroids. CONCLUSION The resistance level can jeopardize the efficiency of chemical adult control in absence of other alternatives and conducts to strongly rely on larval control measures to reduce mosquito burden. Vector control strategies need to evolve to maintain or regain efficacy during epidemics.

Resistance of the stable fly, Stomoxys calcitrans (Diptera: Muscidae), to cypermethrin in outbreak areas in Midwestern Brazil / Resistência da mosca-dos-estábulos, Stomoxys calcitrans (Diptera: Muscidae), à cipermetrina em áreas de surtos no Centro-Oeste brasileiro

Abstract Population explosions of the stable fly (Stomoxys calcitrans) have become a serious concern for livestock producers near sugarcane mills in some regions of Brazil due to the insect's massive reproduction on sugarcane byproducts and waste. Despite the limited efficiency of insecticides for controlling stable fly outbreaks, producers still rely on chemical control to mitigate the alarming infestations in affected areas. This study evaluated the susceptibility of S. calcitrans populations to cypermethrin in the state of Mato Grosso do Sul, Brazil. Stable flies were tested from three field populations and two colonies, established from flies previously collected at sugarcane mills. Wild flies were collected with Nzi traps in areas of sugarcane plantations. Both wild and colonized flies were exposed to eleven concentrations of cypermethrin in impregnated filter paper bioassays. All the populations proved to be resistant to cypermethrin, with resistance factors among field populations ranging from 6.8 to 38.6. The intensive use of insecticides has led to the development of pyrethroid resistance in stable fly populations in the proximities of sugarcane mills in the state of Mato Grosso do Sul.

Resumo Explosões populacionais da mosca-dos-estábulos (Stomoxys calcitrans) tornaram-se uma séria preocupação para a pecuária próxima a usinas de cana-de-açúcar em algumas regiões do Brasil, devido à massiva reprodução da mosca em resíduos e subprodutos do processamento da cana. Apesar da limitada eficiência dos inseticidas no controle dos surtos desta mosca, produtores dependem do controle químico para mitigar as alarmantes infestações nas áreas afetadas. Este estudo objetivou avaliar a suscetibilidade de populações de S. calcitrans a piretroides no estado de Mato Grosso do Sul, Brasil. Moscas-dos-estábulos de três populações de campo e duas colônias, estabelecidas a partir de moscas previamente coletadas em usinas de cana-de-açúcar, foram testadas. Moscas selvagens foram coletadas com armadilhas Nzi em áreas de cana. Moscas coletadas foram expostas a onze concentrações de cipermetrina em bioensaios com papel de filtro impregnado. Todas as populações testadas mostraram-se resistentes ao piretroide, com fatores de resistência variando de 6,8 a 38,6 nas populações de campo. O uso intensivo de inseticidas tem levado ao desenvolvimento de resistência da mosca-dos-estábulos a piretroides em populações próximas a usinas de cana-de-açúcar no estado do Mato Grosso do Sul.

Bio-efficacy, physical integrity, community usage and washing practices of mosquito nets treated with ICON MAXX long-lasting insecticidal treatment in India

BACKGROUND New brands of potential long lasting insecticide nets (LLINs) and LLIN treatment kits require field evaluation before they are used in a vector control programme. OBJECTIVES The aim of this study was to evaluate the bio-efficacy, usage, washing practice and physical integrity of nets treated with LLIN treatment kit, ICON MAXX in a phase III field trial in Odisha state, India. METHODS A total of 300 polyester nets treated with ICON MAXX and 140 polyester nets treated conventionally with lambda-cyhalothrin CS 2.5% ITNs were distributed. The bio-efficacy was evaluated with WHO cone bioassay. The chemical analysis of netting pieces was done at the beginning, after 12 and 36 months of the trial. FINDINGS After one year of distribution of nets, the bioassay showed 100% mortality on both ITNs and ICON MAXX treated nets. At 36 months, the overall pass rate was 58.8% and the mean lambda-cyhalothrin content of LLINs was 34.5 mg ai/m2, showing a loss of 44.4% of the original concentration. CONCLUSION ICON MAXX treated LLIN was found to retain bio-efficacy causing 97% knockdown of Anopheles stephensi up to 30 months and met the WHOPES criteria. However, the desired bio-efficacy was not sustained up to 36 months.

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).

Characterization of the pyrethroid resistance profile of Rhipicephalus (Boophilus) microplus populations from the states of Rio Grande do Sul and Mato Grosso do Sul, Brazil / Caracterização do perfil de resistência do Rhipicephalus (Boophilus) microplus a piretróides em populações dos estados do Rio Grande do Sul e Mato Grosso do Sul, Brasil

Cattle ticks Rhipicephalus (Boophilus) microplus are mainly controlled in Brazil by means of acaricide products, without any official policies in this regard. Acaricides continue to be sold indiscriminately, and this has contributed towards making the problem of resistance widespread, thus making diagnosis and monitoring of tick resistance essential. Here, bioassays (larval packet test) were performed on tick populations from the states of Rio Grande do Sul and Mato Grosso do Sul regarding their susceptibility to cypermethrin, deltamethrin and flumethrin. All the tick samples tested showed resistance to cypermethrin (10) (resistance factor (RF) ranging from 5.6 to 80.3) and deltamethrin (10) (RF ranging from 2.4 to 83.1). Six out of eight populations were resistant to flumethrin (RF ranging from 3.8 to 8.2). PCR molecular analyses did not show any T2134A mutations in the voltage-gated sodium channel gene, in any of the sampled populations. The results from this study highlight the critical status of resistance of the cattle tick to synthetic pyrethroids in the regions studied. Further studies are needed to identify the mechanisms responsible for the resistant phenotypes observed in the bioassays. This was the first detection of flumethrin resistance in Brazil.

O controle do carrapato Rhipicephalus (Boophilus) microplus no Brasil é feito principalmente com produtos acaricidas, sem uma política de controle oficial. A venda destes produtos continua ocorrendo de maneira indiscriminada, o que tem contribuído para generalizar o problema da resistência em todo o país, tornando essencial seu diagnóstico e monitoramento. Bioensaios (teste do pacote de larvas) foram conduzidos com populações de carrapatos do Rio Grande do Sul e Mato Grosso do Sul, com relação à sua suscetibilidade a três princípios ativos piretróides. Todas as amostras analisadas mostraram resistência à cipermetrina (10), com fatores de resistência (FR) de 5,6 a 80,3, e à deltametrina, com FR variando de 2,4 a 83,1. Seis das oito amostras foram resistentes à flumetrina (FR de 3,8 a 8,2). Análises moleculares utilizando PCR demonstraram que em todas as populações amostradas não foram encontrados alelos com a mutação T2134A no gene do canal de sódio controlado por voltagem. Os resultados deste estudo evidenciam a situação crítica da resistência do carrapato bovino ao grupo dos piretróides sintéticos nas regiões estudadas. Novos estudos são necessários para identificar os mecanismos responsáveis pelos fenótipos resistentes observados nos bioensaios. Este é o primeiro relato da resistência a flumetrina no Brasil.

Susceptibility of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) to pyrethroids and their associations in Pernambuco, Brazil / Susceptibilidade de Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) a piretróides e suas associações em Pernambuco, Brasil

The synthetic pyrethroids and their associations have been widely used for controlling Rhipicephalus (Boophilus) microplus. The frequent use of acaricides has been inducing the development of resistance in the tick populations. The aim of this study was to assess the susceptibility of R. (B.) microplus populations to pyrethroids and their associations in the region of Garanhuns, Pernambuco, Brazil. In addition, the level of information among farm owners regarding tick control measures was investigated. Ticks were collected directly from naturally infested dairy cattle in the region and were exposed to pyrethroids and their associations. At the same time, an epidemiological questionnaire was applied with the aim of investigating the level of information among the farmers. The results reported here indicate that R. (B.) microplus populations in the dairy region of Garanhuns show resistance to pyrethroids and their associations, except when the product is associated with piperonyl butoxide. Regarding the results from the epidemiological survey, it was seen that there is a considerable lack of information among the farmers in relation to ixodid control measures. The level of ticks resistance to acaricides varied widely across the region studied. No alternative control programs have been implemented among these farms, thus demonstrating that there is a need for more information relating to the biology and control of R. (B.) microplus.

Os piretróides sintéticos e associações são acaricidas amplamente utilizados para o controle do Rhipicephalus (Boophilus) microplus. O uso frequente destes fármacos tem induzido o surgimento de populações de carrapatos resistentes. Objetivou-se neste estudo avaliar a susceptibilidade de populações de R. (B.) microplus aos piretróides e associações. Além disso, investigou-se o nível de informações dos proprietários com relação ás medidas de controle utilizadas. Os carrapatos foram coletados diretamente de bovinos naturalmente infestados na região leiteira de Garanhuns, Pernambuco e expostos a piretróides e suas associações. Contemporaneamente, foi aplicado um questionário epidemiológico com o objetivo de avaliar o nível de informação dos produtores. Os resultados aqui obtidos demonstram que populações de R. (B.) microplus provenientes da região leiteira de Garanhuns apresentam resistência aos piretróides, exceto quando o produto está associado com butóxido de piperonila. Em se tratando dos resultados referentes ao inquérito epidemiológico, verificou-se considerável falta de informação por parte dos produtores com relação ás medidas de controle de ixodídeos. Assim, verifica-se que o nível de resistência aos acaricidas é muito variável na região estudada e que não há, nas propriedades, a adoção de programas alternativos de controle, demonstrando a necessidade de maiores informações relacionadas á biologia e ao controle do R. (B.) microplus.

Mechanisms of pyrethroid resistance in Haematobia irritans (Muscidae) from Mato Grosso do Sul state, Brazil / Mecanismos de resistência da Haematobia irritans (Muscidae) a piretróides em Mato Grosso do Sul, Brasil

Horn fly resistance to pyrethroid insecticides occurs throughout Brazil, but knowledge about the involved mechanisms is still in an incipient stage. This survey was aimed to identify the mechanisms of horn fly resistance to cypermethrin in Mato Grosso do Sul state, Brazil. Impregnated filter paper bioassays using cypermethrin, synergized or not with piperonyl butoxide (PBO) and triphenyl phosphate (TPP), were conducted from March 2004 to June 2005 in horn fly populations (n = 33) from all over the state. All populations were highly resistant to cypermethrin, with resistance factors (RF) ranging from 89.4 to 1,020.6. Polymerase chain reaction (PCR) assays to detect the knockdown resistance (kdr) mutation also were performed in 16 samples. The kdr mutation was found in 75% of the tested populations, mostly with relatively low frequencies (<20%), and was absent in some highly resistant populations. Addition of TPP did not significantly reduce the LC50 in any population. However, PBO reduced LC50s above 40-fold in all tested populations, resulting in RFs ≤ 10 in most cases. Horn fly resistance to cypermethrin is widespread in the state, being primarily caused by an enhanced activity of P450 mono-oxygenases and secondarily by reduced target site sensitivity.

Resistência da mosca-dos-chifres a inseticidas piretróides ocorre em todo o país, entretanto, o conhecimento sobre os mecanismos envolvidos é ainda incipiente. Este estudo objetivou identificar os mecanismos de resistência desta mosca à cipermetrina em Mato Grosso do Sul. Bioensaios utilizando papéis impregnados com cipermetrina, isoladamente ou sinergizada por butóxido de piperonila (PBO) ou trifenil fosfato (TPP), foram realizados de março∕2004 a junho∕2005 em 33 populações. Todas as populações apresentaram elevada resistência à cipermetrina, com fatores de resistência (FR) variando de 89,4 a 1.020,6. Ensaios de reação em cadeia da polimerase (PCR) visando a detecção de kdr ("knockdown resistance") foram realizados em 16 amostras. A mutação kdr foi detectada em 75% das populações, geralmente em baixas frequências (<20%) e ausente em algumas populações resistentes. A adição de TPP não reduziu significativamente a CL50 em nenhuma população. Entretanto, o PBO reduziu em mais de 40 vezes a CL50 de todas as populações testadas, resultando em FR ≤ 10 na maioria dos casos. Resistência da mosca-dos-chifres à cipermetrina encontra-se disseminada no estado, sendo causada primariamente por um aumento da atividade de P450 mono-oxigenases e secundariamente pela redução da sensibilidade do sítio de ação do inseticida.

Resistance to cypermethrin and amitraz in Rhipicephalus (Boophilus) microplus on the Santa Catarina Plateau, Brazil / Situação da resistência do Rhipicephalus (Boophilus) microplus à cipermetrina e amitraz no Planalto Catarinense, Brasil

With the objective of evaluating Rhipicephalus (Boophilus) microplus resistance to cipermetrina and amitraz, were collected engorged tick females from cattle on 20 farms on the Santa Catarina Plateau, in southern Brazil, between January of 2004 and May of 2006. Were also received 20 groups of engorged R. (B.) microplus females, collected by cattle farmers requesting acaricidal efficacy (AE) testing. Were performed in vitro tests, which consisted of immersing engorged females in cypermethrin (0.015%) and in amitraz (0.025%). An AE >95% was considered indicative of effectiveness. Of the 20 collected groups, 18 (90%) showed cypermethrin resistance and 1 (5%) showed amitraz resistance. Of the 20 received groups, 19 (95%) showed cypermethrin resistance and 2 (10%) showed amitraz resistance. The AE of cypermethrin was found to be >95%, 85-94%, and 55-64%, respectively, in 4 (57.1%), 2 (28.6%), and 1 (14.3%) of 7 reference groups, collected in the 1997-2001 period. The AE of amitraz was >95% in all of those groups. Among the groups of specimens received for analysis in that period, the AE of amitraz was >95% in 6 (85.71%) and 75-84% in 1 (14.28%). R. (B.) microplus resistance to acaricides is increasing on the Santa Catarina Plateau.

Com os objetivos de avaliar a resistência do Rhipicephalus (Boophilus) microplus à cipermetrina e ao amitraz no Planalto Catarinense, no período de janeiro de 2004 a maio de 2006, foram coletadas teleóginas em bovinos de 20 propriedades onde os produtores autorizaram a coleta (amostras por conveniência) e recebidas teleóginas de mais 20 propriedades com objetivo de diagnóstico de eficácia. Os testes in vitro foram realizados por imersão de teleóginas em cipermetrina (0,015%) e amitraz (0,025%). Considerando a eficácia igual ou superior a 95%, das 20 propriedades amostradas, 18 (90%) apresentaram resistência à cipermetrina e uma (5%) ao amitraz. Das 20 propriedades com teleóginas para diagnóstico, 19 (95%) apresentaram resistência à cipermetrina e duas (10%) ao amitraz. No período de agosto de 1997 a dezembro de 2001, das sete amostras coletadas, em quatro (57,1%) a eficácia da cipermetrina foi >95%, em duas (28,6%) entre 85% e 94% e em uma, (14,3%) entre 55% e 64%. A eficácia do amitraz foi >95%, em teleóginas de todas as propriedades. Nas amostras para diagnóstico, seis (85,71%) apresentaram eficácia para o amitraz >95% e uma (14,28%) entre 75% e 84%. A resistência desse ixodídeo aos carrapaticidas testados é crescente no Planalto Catarinense.

Acaricide resistance mechanisms in Rhipicephalus (Boophilus) microplus / Mecanismos de resistência aos acaricidas em Rhipicephalus (Boophilus) microplus

Acaricide resistance has become widespread in countries where cattle ticks, Rhipicephalus (Boophilus) microplus, are a problem. Resistance arises through genetic changes in a cattle tick population that causes modifications to the target site, increased metabolism or sequestration of the acaricide, or reduced ability of the acaricide to penetrate through the outer protective layers of the tick’s body. We review the molecular and biochemical mechanisms of acaricide resistance that have been shown to be functional in R. (B.) microplus. From a mechanistic point of view, resistance to pyrethroids has been characterized to a greater degree than any other acaricide class. Although a great deal of research has gone into discovery of the mechanisms that cause organophosphate resistance, very little is defined at the molecular level and organophosphate resistance seems to be maintained through a complex and multifactorial process. The resistance mechanisms for other acaricides are less well understood. The target sites of fipronil and the macrocyclic lactones are known and resistance mechanism studies are in the early stages. The target site of amitraz has not been definitively identified and this is hampering mechanistic studies on this acaricide.

A resistência aos acaricidas tornou-se amplamente difundida nos países onde os carrapatos bovinos, Rhipicephalus .Boophilus. microplus, são um problema. A resistência surge por meio de alterações genéticas em umapopulação de carrapatos que causam modificações no local de ação, aumento do metabolismo ou sequestro do acaricida, ou ainda redução na capacidade do acaricida em penetrar através das camadas protetoras do corpo do carrapato. Neste artigo, foram revisados os mecanismos moleculares e bioquímicos da resistência aos acaricidas que ocorrem em R. (B.) microplus. A partir de um ponto de vista dos mecanismos envolvidos, a resistência aos piretróides tem sido caracterizada em maior grau do que em qualquer outra classe de acaricida. Embora uma grande quantidade de pesquisas têm sido direcionada para a descoberta de mecanismos que causam resistência aos organofosforados, muito pouco é conhecido ao nível molecular, e essa resistência parece ser mantida por intermédio de um processo multifatorial e complexo. Os mecanismos de resistência para os demais acaricidas são bem menos compreendidos. Os alvos de ação do fipronil e das lactonas macrocíclicas são conhecidos, e os estudos dos mecanismos de ação envolvidos estão ainda em estágios iniciais. O alvo de ação do amitraz ainda não foi definitivamente identificado, e isso é limitante aos estudos dos mecanismos envolvidos na resistência a esse acaricida.

-Tocopherol ameliorates cypermethrin-induced toxicity and oxidative stress in the nematode Caenorhabdtis elegans.

Oxidative stress and other effects induced by cypermethrin (CYP, 15 mM) and their amelioration by -tocopherol (400 M) was studied in the nematode Caenorhabditis elegans. The worms exposed for 4 h to CYP showed increased levels of reactive oxygen species (46%), H2O2 (37%) and protein carbonyls (29%), accompanied by decreased lifespan and brood size. However, exposure to both CYP and a-tocopherol resulted in diminution of above alterations with the worms exhibiting relatively lower levels of ROS (30%), H2O2 (15%), protein carbonyls (14%), altered antioxidant enzyme activities and normal lifespan and brood size. The results suggest that CYP induces oxidative stress in C. elegans and the strategy of intervention with -tocopherol could be exploited to offset this induced oxidative stress.

Field evaluation of ZeroFly® - an insecticide incorporated plastic sheeting against malaria vectors & its impact on malaria transmission in tribal area of northern Orissa.

Background & objectives: Insecticide incorporated plastic sheeting is a new technology to control mosquitoes in emergency shelter places and also temporary habitations in different locations. Therefore, field studies were conducted to assess the efficacy of ZeroFly® plastic sheeting treated with deltamethrin on prevailing disease vectors Anopheles culicifacies and An. fluviatilis and its impact on malaria transmission in one of the highly endemic areas of Orissa. Methods: The study was conducted in Birkera block of Sundargarh district, Orissa state. The study area comprised 3 villages, which were randomized as ZeroFly plastic sheet, untreated plastic sheet and no sheet area. ZeroFly plastic sheets and untreated plastic sheets were fixed in study and control villages respectively covering all the rooms in each household. Longitudinal studies were conducted on the bioefficacy with the help of cone bioassays, monitoring of the mosquito density through hand catch, floor sheet and exit trap collections and fortnightly domiciliary active surveillance in all the study villages. Results: In ZeroFly plastic sheeting area, there was a significant reduction of 84.7 per cent in the entry rate of total mosquitoes in comparison to pre-intervention phase. There was 56.2 per cent immediate mortality in total mosquitoes in houses with ZeroFly sheeting. The overall feeding success rate of mosquitoes in the trial village was only 12.5 per cent in comparison to 49.7 and 51.1 per cent in villages with untreated plastic sheet and no sheet respectively. There was a significant reduction of 65.0 and 70.5 per cent in malaria incidence in ZeroFly plastic sheeting area as compared to untreated plastic sheet and no sheet area respectively. Interpretation & conclusions: Our study showed that introduction of ZeroFly plastic sheets in a community-based intervention programme is operationally feasible to contain malaria especially in the high transmission difficult areas.

Effect of cyhalothrin on Ehrlich tumor growth and macrophage activity in mice

Cyhalothrin, a pyrethroid insecticide, induces stress-like symptoms, increases c-fos immunoreactivity in the paraventricular nucleus of the hypothalamus, and decreases innate immune responses in laboratory animals. Macrophages are key elements in cellular immune responses and operate at the tumor-host interface. This study investigated the relationship among cyhalothrin effects on Ehrlich tumor growth, serum corticosterone levels and peritoneal macrophage activity in mice. Three experiments were done with 10 experimental (single gavage administration of 3.0 mg/kg cyhalothrin daily for 7 days) and 10 control (single gavage administration of 1.0 mL/kg vehicle of cyhalothrin preparation daily for 7 days) isogenic BALB/c mice in each experiment. Cyhalothrin i) increased Ehrlich ascitic tumor growth after ip administration of 5.0 x 106 tumor cells, i.e., ascitic fluid volume (control = 1.97 ± 0.39 mL and experimental = 2.71 ± 0.92 mL; P < 0.05), concentration of tumor cells/mL in the ascitic fluid (control = 111.95 ± 16.73 x 106 and experimental = 144.60 ± 33.18 x 106; P < 0.05), and total number of tumor cells in the ascitic fluid (control = 226.91 ± 43.22 x 106 and experimental = 349.40 ± 106.38 x 106; P < 0.05); ii) increased serum corticosterone levels (control = 200.0 ± 48.3 ng/mL and experimental = 420.0 ± 75.5 ng/mL; P < 0.05), and iii) decreased the intensity of macrophage phagocytosis (control = 132.3 ± 19.7 and experimental = 116.2 ± 4.6; P < 0.05) and oxidative burst (control = 173.7 ± 40.8 and experimental= 99.58 ± 41.7; P < 0.05) in vitro in the presence of Staphylococcus aureus. These data provide evidence that cyhalothrin simultaneously alters host resistance to Ehrlich tumor growth, hypothalamic-pituitary-adrenocortical (HPA) axis function, and peritoneal macrophage activity. The results are discussed in terms of data suggesting a link between stress, HPA axis activation and resistance to tumor growth.

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.

Mixed-function oxidases and esterases associated with cross-resistance between DDT and lambda-cyhalothrin in Anopheles darlingi Root 1926 populations from Colombia

In order to establish the insecticide susceptibility status for Anopheles darlingi in Colombia, and as part of the National Network on Insecticide Resistance Surveillance, five populations of insects from three Colombian states were evaluated. Standardised WHO and CDC bottle bioassays, in addition to microplate biochemical assays, were conducted. Populations with mortality rates below 80 percent in the bioassays were considered resistant. All field populations were susceptible to deltamethrin, permethrin, malathion and fenitrothion. Resistance to lambda-cyhalothrin and DDT was detected in the Amé-Beté population using both bioassay methods with mortality rates of 65-75 percent. Enzyme levels related to insecticide resistance, including mixed function oxidases (MFO), non-specific esterases (NSE), glutathione S-transferases and modified acetylcholinesterase were evaluated in all populations and compared with a susceptible natural strain. Only mosquitoes from Amé-Beté presented significantly increased levels of both MFO and NSE, consistent with the low mortalities found in this population. The continued use of lambda-cyhalothrin for An. darlingi control in this locality has resulted in a natural resistance to this insecticide. In addition, DDT resistance is still present in this population, although this insecticide has not been used in Colombia since 1992. Increased metabolism through MFO and NSE may be involved in cross-resistance between lambda-cyhalothrin and DDT, although kdr-type nerve insensitivity cannot be discarded as a possible hypothesis. Additional research, including development of a kdr specific assay for An. darlingi should be conducted in future studies. Our data demonstrates the urgent need to develop local insecticide resistance management and surveillance programs throughout Colombia.

Stability and wash resistance of local made mosquito bednets and detergents treated with pyrethroids against susceptible strain of malaria vector anopheles stephensi

We aimed to evaluate different fibres of bednets impregnated with various pyrethroids. The stability of insecticide on the bednet was measured using different methods of washings as well as local made detergents. The entire test was carried out according to the WHO-recommended methods. In addition, the impact of the numbers of washes on the stability of the insecticides was determined. Permethrin 10% [EC], deltamethrin 10% [SC], lambdacyhalothrin 2.5% [CS] and cyfluthrin 5% [EW] were used at the recommended dosages. Three different local detergents were used. Two kinds of washing methods [shaking, no shaking] were used and in each method four kinds of washings, i.e. no wash, one wash, two washes and three washes was done. The main malaria vectors, Anopheles stephensi, which is susceptible to all insecticides [BEECH strain], was tested with impregnated bednets in 3 minutes exposure time and the mortality was measured after 24 hours recovery period. Knock-down was measured as well using appropriate statistical methods. Lambdacyhalothrin has saved its insecticidal impact after being washed, whereas, deltamethrin has lost its activity faster than other insecticides. Tow other insecticides had moderate effect. Golnar soap detergent has least effect on the durability of insecticides, but the Shoma had the most. Whit increasing the times of washing, insecticidal effects was decreased, but shaking had no influence on the decreasing of the quality of insecticidal impact. Results will be useful for local people who wish to use pyrethroid-impregnated bednets with their own local made detergent and bednets

Control of aggregated populations of the eye fly Siphunculina funicola (diptera: chloropidae) using pyrethroid aerosols.

Three commercially available insecticide aerosol compositions containing cyphenothrin, imiprothrin, cypermethrin, d-tetramethrin and permethrin, in various combinations and concentrations were evaluated for control of the oriental eye flies Siphunculina funicola (de Meijere) from their aggregation substrates in two villages in the Chon Buri Province, Thailand. Each aerosol produced almost complete kill of eye flies on most of the treated substrates. Time to reoccupation of each treated substrate was determined at 24, 48 and 72 hours post-treatment. All 3 aerosol preparations killed eye flies on most of the treated substrates and kept most of the substrates free of eye flies for at least 48-72 hours. The effectiveness and longevity of the aerosols varied depending on the type and location of the substrate. Smooth, hairy and polished substrates did not intercept sufficient insecticide; therefore providing low residual activity compared to porous and coated surfaces. Aerosols applied to substrates located at higher levels gave good initial kill, but reoccupation of these substrates by flies occurred relatively quickly (within 24-48 hours) following treatment. Attempts should be made to discharge aerosol flumes closer than 1 m from the aggregation substrates.

Control of mosquito vectors of tropical infectious diseases: (1) bioefficacy of mosquito coils containing several pyrethroids and a synergist.

The bioefficacy of mosquito coils containing several pyrethroids were tested in a 25 m3 room against Culex pipiens quinquefasciatus, Aedes aegypti and Anopheles dirus. The test results were compared with tests against Culex pipiens pallens in Japan. Based on the KT50 values (the 50% knockdown time) of mosquito coils containing dl, d-T80-allethrin, d, d-T-prallethrin and methoxymethyl-tetrafluorobenzyl tetramethyl-cyclopropanecarboxylate (K-3050) at doses of 0.05-0.5% (w/w) with or without a synergist, the pyrethroid susceptibility of the four mosquito species was as follows: Cx. p. quinquefasciatus was several times more tolerant to pyrethroids than Cx. p. pallens, Ae. aegypti was a further several times more tolerant than Cx. p. quinquefasciatus, and An. dirus was more susceptible than Cx. p. pallens (KT50 value: about half of Cx. p. pallens). The order of their susceptibilities is common for pyrethroids. Mosquito coils containing d, d-T-prallethrin and K-3050 at doses of 0.05-0.2% (w/w) and N-(2-ethylhexyl)bicycle-[2,2,1]-hept-5-ene-2,3-dicarboxyimide as a synergist at a ratio of 2 times the active ingredient were highly effective against Ae. aegypti, the most important mosquito vector for dengue fever.