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Background: Chagas disease, a significant global health concern rooted in social inequalities and political oversights, remains a challenging public health issue impacting millions worldwide. The increasing detection of triatomines, the vectors of Chagas disease, in urban areas complicates the situation., Methods: This study investigated the incidence of Rhodnius neglectus in the urban areas of Jaboticabal, São Paulo, Brazil, following several reports and previous collections of triatomines in the city. An educational approach was adopted, distributing informational materials and engaging the community through social networks to generate notifications that would enable the identification of triatomines. Specimens were collected using various methods, including passive surveillance actions, light traps, and active searches in palm trees., Results: Rhodnius neglectus was found in urban areas, invading homes in Jaboticabal, and was identified in palm trees. The educational approach led to the collection of 93 triatomines. Colonization was observed in a residence, with eggs, nymphs, and a concerning record of blood-feeding on a resident child. The houses where specimens were captured often had nearby palm trees with birds and nests, facilitating the passive transport of these insects and increasing the risk of invasion due to light attraction. No triatomines infected with Trypanosoma cruzi were identified., Conclusions: These findings emphasize the need for preventive measures to reduce the prevalence of R. neglectus in urban environments. The data elucidate the occurrence of R. neglectus in the city of Jaboticabal, associated with its potential behavioral adaptation in urban environments, underscoring the need for innovative control strategies.

Background: Among the 18 genera of the Triatominae subfamily, three stand out for their diversity and epidemiological importance: Triatoma, Panstrongylus, and Rhodnius. Rhodnius includes 21 species that can transmit Trypanosoma cruzi (the etiological agent of Chagas disease, also known as American trypanosomiasis) and Trypanosoma rangeli. The Rhodnius prolixus complex comprises seven species, including Rhodnius marabaensis, Rhodnius prolixus, and Rhodnius robustus, which occur in the northern region of Brazil. Since both adults and immatures can carry T. cruzi, in this study the five nymphal instars of the three species mentioned were dorsally characterized. Methods: Using microscopy, morphometrics, and geometric morphometrics, the present work measures and describes the morphological characters of the five nymphal instars of R. marabaensis, R. prolixus, and R. robustus. Results: The study enabled the characterization of all five nymphal instars, as well as the distinction between the three species in each of their instars. Conclusions: The morphological, morphometrics of the head, thorax, and abdomen and geometric morphometrics studies of the head enabled the specific distinction of these three species in all five instars. [ABSTRACT FROM AUTHOR]

Background: In the Amazon region, several species of triatomines occur in the natural environments. Among them, species of the genus Rhodnius are a risk to human populations due to their high rates of infection with Trypanosoma cruzi. The aim of this study was to identify the T. cruzi genotypes in Rhodnius specimens and their relationship with sylvatic hosts from different environments in the Brazilian Amazon. Methods: A total of 492 triatomines were collected from the municipalities of Monte Negro, Rondônia state, and Humaitá, Amazonas state, 382 of them being nymphs and 110 adults. Genotyping of T. cruzi in six discrete typing units (DTUs) was performed using conventional multilocus PCR. The triatomines that were positive for T. cruzi and engorged with blood were also targeted for amplification of the cytochrome B (cytB) gene to identify bloodmeal sources. Results: Of the 162 positive samples, the identified DTUs were TcI (87.65%) and TcIV (12.35%). It was observed that 102 specimens were engorged with a variety of bloodmeals. Triatomines infected with TcI were associated with DNA of all identified vertebrates, except Plecturocebus brunneus. TcIV was detected in triatomines that fed on Coendou prehensilis , Didelphis marsupialis , Mabuya nigropunctata , P. brunneus , Pithecia irrorata , Sapajus apella , and Tamandua tetradactyla. Conclusion: Results highlight the need to understand the patterns of T. cruzi genotypes in Rhodnius spp. and their association with sylvatic hosts to better elucidate their role in the transmission of Chagas disease in the Amazon region.

Triatominae are associated with various Brazilian habitats, including bird nests, animal burrows, and peridomestic structures. Despite extensive studies on triatomines in domiciliary environments in Ceará, Brazil, there has been limited research on their presence in the wild. This study focuses on the municipality of Morrinhos in Ceará, which is characterized by a Caatinga biome and riparian forests along the Acaraú River. During the study, a total of 185 nests of Pseudoseisura cristata were analyzed in rural locations and triatomines were collected in 26 nests from 12 different locations. A total of 117 triatomines was collected, belonging to three species: Psammolestes tertius , Rhodnius nasutus , and Triatoma pseudomaculata . Rhodnius nasutus was the only species found in a nest inhabited by Didelphis albiventris . Nests of P. cristata serve as shelters for various animals, providing an ideal environment for triatomines to establish colonies due to their proximity to food sources and their generalist feeding habits. The incorporation of anthropogenic materials by birds in nest construction can indirectly affect the presence of other animals that may serve as food sources for triatomines. Understanding the interactions between triatomines, birds, and their habitats is essential for assessing the risks associated with triatomine infestation in wild ecosystems.

Background: An external quality assessment on the identification of triatomines within the laboratory network in the state of Rondônia., Methods: Seven laboratories participated in this evaluation. Each was provided with support materials and nine insects from the Hemiptera order for identification., Results: All samples were accurately identified at the species level. However, correct sex identification was achieved for only 79% of the samples. The most significant challenges were encountered in determining the sex of predators, phytophagous species, Rhodnius robustus, and Rhodnius pictipes., Conclusions: The identified shortcomings can inform enhancements in vector control programs for Chagas disease.

Background: Triatomines infest atypical public spaces in the Western Amazon., Methods: Frequent visitors to these spaces captured the insects in the state of Acre, Brazil (Rio Branco and Cruzeiro do Sul)., Results: Six insects were found in a penitentiary, a church, a school, a university, a hospital, and a health center. Five of the insects were adults (three positive for Trypanosoma cruzi) and one was a nymph., Conclusions: This is the first report of triatomine occurrence in schools or churches. These data are important for implementing surveillance strategies and alerting individuals about possible changes in Chagas disease transmission dynamics.

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Abstract: Among the states that comprise the legally defined Amazon region of Brazil, Pará has presented the highest occurrences of acute cases of Chagas disease over the last two decades. These cases have been attributed to consumption of fruits from native palm trees. In surveys in rural and wild areas of the municipality of Oriximiná, Pará, triatomine fauna, their main ecotopes and the infection rate due to Trypanosoma cruzi were identified using active and passive search methods: manual capture and Noireau traps, respectively. A total of 582 ecotopes were surveyed using 1496 Noireau traps. Out of 442 specimens collected, 289 were identified as Rhodnius robustus and 153 as Rhodnius pictipes. The infection rate caused by T. cruzi was 17.4%. The food sources of the triatomines were found to be birds, hemolymph, horses, and rodents. The association between R. robustus and inajá palm trees (Attalea marita), which are abundant in rural areas, was confirmed. On the other hand, R. pictipes is found in several palm tree species, such as inajá (A. marita), mucajá (Acrocomia aculeata), murumuru (Astrocaryum murumuru) and patauá (Oenocarpus bataua), and in bromeliads in wild areas. These occurrences of triatomine species in regions with or without T. cruzi infection, in the vicinity of the main settlement of the municipality, suggest that there is a need for entomological and epidemiological surveillance in this region. [Copyright &y& Elsevier]

Abstract: This work evaluated the occurrence and genetic structure of Rhodnius nasutus sampled in two sites using morphometry and microsatellites. These sites, presented distinct abiotic features and palm trees: (i) nine Attalea speciosa palm trees, so called babaçu, were sampled from the Meruoca Mountain Ridge, a sloping region of reminiscent forest in the state of Ceará, Brazil, and (ii) 17 Copernicia prunifera palm trees, so called carnaúba, were sampled in the scrub savanna region (Sobral district) that surrounds the mountain ridge. Of the twenty-six palm trees dissected, 70.6% of carnauba and 88.9% of babaçu were infested by R. nasutus. The micro-climatic data where R. nasutus were sheltered demonstrated that the babaçu and carnaúba palm trees presented significant differences (p <0.05) in relation to the external environment, except for temperature and relative humidity regulation, suggesting that the architecture of the babaçu crown keeps a more stable micro-environment. The morphometric studies of the F1 generation demonstrated that insects from the babaçu (A. speciosa) were significantly larger (p =0.000) than those collected in carnaúba (C. prunifera) palm trees. Also, microsatellite analysis demonstrated a high genetic differentiation between the two groups of R. nasutus (R st =−0.77). Our results suggest that the difference in size between the populations is probably related to an incipient process of genetic drift in populations associated to each palm tree, probably also driven by the different climatic features observed in these micro-environments. [Copyright &y& Elsevier]

AbstractChagas disease is caused by the parasitic protozoan Trypanosoma cruziand affects about 15 million people in the Americas. In the Brazilian Amazon Basin, the disease is enzootic, with bugs from the genus Rhodniusserving as the main vectors, while in the northeastern region, mainly the genus Triatomais naturally infected with T. cruzi. Oral infections appear sporadically in some regions of Brazil. Even though the illness is typically present in Latin America, autochthonous cases have been reported in the United States, mainly in the south of the country. The Triatominae subfamily comprises a large number of insect species that are potential vectors of Chagas disease. Triatomines are hematophagous insects, ingesting blood in all life stages. Since the insects play an important role in parasite selection, they also influence the geographical distribution of T. cruzi. The globalization process is one of the most important elements influencing the outcome of Chagas diseases in Brazil. This article outlines the status of reemerging Chagas disease in different regions of Brazil, the distribution of its vectors, and the consequences of global climate changes with respect to interactions of the pathogen with triatomines. [ABSTRACT FROM AUTHOR]

Background: Chagas disease (CD) is caused by the flagellate protozoan Trypanosoma cruzi and can be carried by different species of triatomines, including Rhodnius neglectus, which is wild, well distributed in Brazil, and has formed colonies in palm trees located in urban areas of municipalities in the state of São Paulo. Chemical control has been routinely used to reduce population density, but each year, there has been an increase in species dispersion and density. This study aimed to evaluate the susceptibility of insects to insecticides used in control., Methods: The reference population was collected from Araçatuba municipality, Nilce Maia. Dilutions of deltamethrin were prepared and applied to the back of the first-stage nymphs, which were biologically synchronized. The control group received pure acetone only. Mortality was assessed after 72 h., Results: The mortality rate with respect to diagnostic dose was 100%. The susceptibility profile observed for this population showed RR50 ranging from 1.76 to 3.632., Conclusions: The populations were susceptible to the insecticides tested. It is possible that the insecticide residual effect on this ecotope has decreased the lifespan, and controlling failures may be the cause of recolonization in this environment.

The present study aimed to identify the triatomine species and evaluate Trypanosoma cruzi infection in insects captured in endemic areas of Pará State, Brazil. Triatomines were captured in nine rural communities in the municipality of São Domingos do Capim in August, September and December 2014 using active searches and Malaise and Noireau traps. Additionally, from 2014 to 2018, residents and community health agents submitted captured triatomines to the study team. The analysis of T. cruzi infection in the insects was performed by direct parasitological examination and nested-PCR. A total of 225 triatomines were captured and identified: Rhodnius robustus (n = 111), Rhodnius pictipes (n = 54), Panstrongylus geniculatus (n = 44), Eratyrus mucronatus (n = 11), Panstrongylus lignarius (n = 4), and Panstrongylus rufotuberculatus (n = 1). Direct parasitological examination was performed in 27 living triatomines R. robustus (n = 14), P. geniculatus (n = 7) and R. pictipes (n = 6) and metacyclic trypomastigote forms similar to those of T. cruzi were observed in 66.6% (18/27) samples. Of 174 samples analysed by nested-PCR, 81.6% were positive for T. cruzi DNA: R. robustus (84.7%; 72/85), R. pictipes (84.1%; 37/44), P. geniculatus (69.4%; 25/36), P. lignarius (100%; 4/4), E. mucronatus (75%; 3/4) and P. rufotuberculatus (100%; 1/1). R. robustus, R. pictipes and P. geniculatus were the main vectors of T. cruzi in the studied areas; however, the detection of infections in P. lignarius, E. mucronatus and P. rufotuberculatus indicated that these species can also act as potential vectors of T. cruzi in the study areas., (© 2021 Wiley-VCH GmbH.)

In the Amazon region, Trypanosoma cruzi transmission cycles involve a great diversity of Triatominae vectors and mammal reservoirs. Some Rhodnius spp. mainly inhabit palm trees that act as microhabitats for hosts and vectors. The current study aimed to describe aspects of the bio-ecology of the vectors and reservoirs of T. cruzi in relation to human populations resident near areas with large quantities of palm trees, in rural, peri-urban and urban collection environments, located in the Western Brazilian Amazon. Rhodnius pictipes and Didelphis marsupialis were respectively the most predominant vector and reservoir, with rates of 71% for R. pictipes and 96.5% for D. marsupialis. The vast majority of T. cruzi isolates clustered with TcI. The most prevalent haplotype was TcI COII1 (69.7%). Mauritia flexuosa and Attalea phalerata were the main ecological indicators of infestation by triatomines. Birds were the most common food source (27,71%). T. cruzi isolated from R. robustus has the haplotype HUM-13, previously detected in a chronic Chagas patient living in the same area. Our results demonstrate the relevance of this study, with the occurrence of elevated infection rates in animals, and suggest the importance of the Amazon zones where there is a risk of infection in humans., (© 2021 The Royal Entomological Society.)

Introduction: Triatomines are insect vectors of Trypanosoma cruzi, the etiological agent of Chagas disease., Methods: Triatomines were collected from households and by dissecting palm trees in the peri-urban areas of Cruzeiro do Sul (Acre); they were identified using a specific key and via genital analyses. Trypanosomatid infection was determined through microscopy and polymerase chain reaction., Results: In total, 116 triatomines of the species Eratyrus mucronatus, Rhodnius pictipes, R. stali, and R. montenegrensis were collected, of which 13.8% were positive for T. cruzi., Conclusions: Four species of triatomines presented an infection rate above 13% in the Boca do Moa community.

Introduction: Rhodnius domesticus Neiva & Pinto, 1923 is a rare sylvatic triatomine endemic to the Atlantic Forest, with one known record for Espírito Santo (ES), Brazil from 1969. We present here its rediscovery in ES, 42 years after its first record., Methods: In January 2011, a triatomine specimen was collected from a rural area of the municipality of Santa Teresa, ES., Results: We confirmed this as a new record of R. domesticus in the Baixo Caldeirão locality., Conclusions: This finding supports the possibility of a wild population of R. domesticus in the mountainous region of the Atlantic forest of ES.

Introduction: Triatomines are hematophagous insects that are important to public health since they are the vectors of American Trypanosomiasis. The objective of this study was to describe the occurrence of triatomines in homes in Cruzeiro do Sul, Acre, Brazil., Methods: The specimens were collected by an active search inside homes and also by a passive search by the residents., Results: A total of 55 triatomines were captured comprising of 5 species each of the genera Rhodnius, Eratyrus, and Panstrongylus. No colonies were detected, ruling out the possibility of domiciliation., Conclusions: Information on regional epidemiological dynamics contributes to the prevention and control of disease.

Background: Rhodnius montenegrensis (Triatominae), a potential vector of Chagas disease, was described after R. robustus-like bugs from southwestern Amazonia. Mitochondrial cytb sequence near-identity with sympatric R. robustus (genotype II) raised doubts about the taxonomic status of R. montenegrensis, but comparative studies have reported fairly clear morphological and genetic differences between R. montenegrensis and laboratory stocks identified as R. robustus. Here, we use a transcriptome-based approach to investigate this apparent paradox., Results: We retrieved publicly-available transcriptome sequence-reads from R. montenegrensis and from the R. robustus stocks used as the taxonomic benchmark in comparative studies. We (i) aligned transcriptome sequence-reads to mitochondrial (cytb) and nuclear (ITS2, D2-28S and AmpG) query sequences (47 overall) from members of the R. prolixus-R. robustus cryptic-species complex and related taxa; (ii) computed breadth- and depth-coverage for the 259 consensus sequences generated by these alignments; and, for each locus, (iii) appraised query sequences and full-breadth-coverage consensus sequences in terms of nucleotide-sequence polymorphism and phylogenetic relations. We found evidence confirming that R. montenegrensis and R. robustus genotype II are genetically indistinguishable and, hence, implying that they are, in all likelihood, the same species. Furthermore, we found compelling genetic evidence that the benchmark 'R. robustus' stocks used in R. montenegrensis description and in later transcriptome-based comparisons are in fact R. prolixus, although likely mixed to some degree with R. robustus (probably genotype II, a.k.a. R. montenegrensis)., Conclusions: We illustrate how public-domain genetic/transcriptomic data can help address challenging issues in disease-vector systematics. In our case-study, taxonomic confusion apparently stemmed from the misinterpretation of sequence-data analyses and misidentification of taxonomic-benchmark stocks. More generally, and together with previous reports of mixed and/or misidentified Rhodnius spp. laboratory colonies, our results call into question the conclusions of many studies (on morphology, genetics, physiology, behavior, bionomics or interactions with microorganisms including trypanosomes) based on non-genotyped 'R. prolixus' or 'R. robustus' stocks. Correct species identification is a prerequisite for investigating the factors that underlie the physiological, behavioral or ecological differences between primary domestic vectors of Chagas disease, such as R. prolixus, and their sylvatic, medically less-relevant relatives such as R. robustus (s.l.) including R. montenegrensis.

The biological behaviour of 23 Trypanosoma cruzi isolates in Swiss mice was compared. Nineteen isolates were obtained from patients in the acute phase of Chagas disease (13), sylvatic reservoir hosts (Didelphis marsupialis) (3), and triatomine bugs (Rhodnius robustus) (3) from four regions of the State of Amazonas (AM). Four isolates were obtained from chronic chagasic patients in the State of Paraná (PR): three autochthones, and one allochthone from the State of Minas Gerais. Only one isolate was unable to infect the mice. The AM and PR isolates showed the largest number of significant differences from each other. The former had lower mean values in the pre-patent (5.4 days) and patent (4.6 days) periods (PP), with the parasitaemia (Pmax) reaching a peak of 9.9×10(4) blood trypomastigotes (BT)/mL of blood by the 7th day following inoculation. The AM isolates also had higher positivity to fresh-blood examination (FBE) (84.1%) compared to haemoculture (HC) (58.7%) and polymerase chain reaction (PCR) (33.3%), in addition to higher mortality (2.9%). The PR isolates had higher values for PP (18.5 days) and Pmax (99.9×10(4)BT/mL) as well as higher positivity to FBE (87.2%), HC (100%), and PCR (83.3%). The correlations between the biological behaviour of the T. cruzi isolates and the clinical and epidemiological characteristics of Chagas disease are discussed., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Introduction: Chagas' disease is not considered an endemic disease in the State of Maranhão. However, entomological surveys showed high natural vector infection indices and acute cases have been identified in the last few decades. This study aimed to analyze the social, demographic and environmental conditions involved in transmission., Methods: The study describes acute Chagas' disease identified from 1994 to 2008. Information regarding the cases was obtained from the Information System for Notifiable Diseases (Sinan), National Health Foundation (Funasa) database and medical records. Entomological surveys were conducted from 2002., Results: Data analysis identified 32 cases from 17 municipal districts, with 84.4% of patients from rural areas. The disease was more frequent in men (67%). The most frequent occupation was student, 38.9% of cases, followed by farmer and hunter, 27.8%., Conclusions: The study suggests that transmission was vectorial and occurred in the wild or peridomicile.

Introduction: A seroepidemiological and clinical study was conducted on 152 autochthonous individuals living in the district of Barcelos, State of Amazonas, to evaluate the seroprevalence of Chagas infection and morbidity of Chagas disease., Methods: The serological tests used were indirect immunofluorescence, conventional and recombinant ELISA and immunoblot (Tesa-blot). Thirty-eight patients were considered seropositive; 31 were considered serodoubtful; and 83 were considered seronegative. The 38 seropositive cases were paired with 38 seronegative controls of the same age and sex, and underwent epidemiological and clinical evaluations, electrocardiograms and echocardiograms. Twenty-nine pairs underwent radiological examinations of the esophagus., Results: Seropositivity was 19.9 times more frequent among workers gathering plant materials from the forests and 10.4 times more frequent among piassaba gatherers. Eighty six point seven percent of the seropositive individuals recognized the genus Rhodnius as the local vector, while only 34.2% of the seronegative individuals recognized this. The EKG was abnormal in 36.8% of the seropositive individuals and in 21.5% of the seronegative individuals, while the echocardiogram showed abnormalities in 31.6% of the seropositive and 18.4% of the seronegative individuals. Precordialgia and palpitation were more frequent among the seropositive individuals. Clinical evaluation on the digestive system and X-ray on the esophagus did not show significant abnormalities., Conclusions: Chagas disease in the study region can be considered to be an occupational disease.

This study has as purpose to accomplish diagnosis of the situation of the Chagas' disease in the old settlements of Porto Letícia, composing picture of the occurrence in the Pontal of Paranapanema. Assisting the comparison with recent groupings the study behaved the accomplishment of triatomines research, of knowledge of the vector tends in its view notification and of the serum- prevalence of the disease. Porto Letícia constitung the oldest settlement congregated a population of young structure, similar to observed her in the settlement Nova Esperança III. With coming inhabitants of endemic areas Porto Letícia presented the highest (5.6%) serum-reagents proportion. Two residences presented infestation in this place tends as counterpoint the triatomines absence in Nova Esperança III and of 24 homes infested in the re-settlements Fazenda Buritis. Low disposition levels for notification were observed in the groupings. Considering the situation of risk of trasmission of the disease the study reaffirms the importance of the notification on the part of the population suggesting an increase of the educational activities.

RAPD analysis and sequences of the mini-exon and ribosomal genes show that Trypanosoma cruzi can be clustered into two phylogenetic groups-T. cruzi I and II. Herein, the Representational Difference Analysis (RDA) method was used, providing new targets specific for each group. After three rounds of RDA hybridizing F strain (tester) with Y strain (driver) and vice-versa, an F-specific (F#30) and Y-specific (Y#22) clone were obtained specifically recognizing isolates from Amazonas (T. cruzi I) and Piauí (T. cruzi II). These segments corresponded to an unspecified protein (F#30) and a trans-sialidase (Y#22). Analysis of the F#30 sequence in T. cruzi I, T. cruzi II and zymodeme 3 samples displayed negligible specific differences that distinguished each group. In addition this F#30 gene has great potential as a hybrid marker.

This study presents the first genetic characterization of five Trypanosoma rangeli isolates from Minas Gerais, in the southeast of Brazil and their comparison with Colombian populations by minicircle classification, RAPD-PCR and LSSP-PCR analyses. Our results demonstrated a homogenous T. rangeli population circulating among Didelphis albiventris as reservoir host in Brazil while heterogeneous populations were found in different regions of Colombia. KP1(+) minicircles were found in 100% isolates from Brazil and in 36.4% of the Colombian samples, whereas the KP2 and KP3 minicircles were detected in both groups. RAPD-PCR and LSSP-PCR profiles revealed a polymorphism within KP1(+) and KP1(-) T. rangeli populations and allowed the division of T. rangeli in two branches. The Brazilian KP1(+) isolates were more homogenous than the KP1(+) isolates from Colombia. The RAPD-PCR were entirely consistent with the distribution of KP1 minicircles while those obtained by LSSP-PCR were associated in 88.9% and 71.4% with KP1(+) and KP1(-) populations, respectively.

This study had the aim of assessing the characteristics of triatomine infestation in human dwellings in rural settlement and resettlement areas with regard to the time when infestation began. We analyzed data relating to 48 triatomine surveys carried out in 105 settlement areas and six resettlement areas in the region of Pontal do Paranapanema between January 1984 and June 2005. Among the 16 surveys in settlement areas, seven (43.8%) had positive findings, all of them in communities established eight or more years previously. Among the 32 surveys in resettlement areas, 23 (71.9%) had positive findings, all of them in communities established for periods shorter than eight years. Since the inhabitants of such communities frequently move, the need for constant vigilance to detect any cases of infestation by vector triatomines in new settlements cannot be overemphasized.

Resistance levels to insecticides used in control of Chagas Disease vectors were assessed in two species of bugs (Hemiptera: Reduviidae): Triatoma infestans (Klug) from Brazil and Rhodnius prolixus Stål from Venezuela. The resistance ratios, compared to susceptible laboratory strains, were determined by topical application bioassays. The T. infestans PA strain exhibited resistance ratios of 7x to deltamethrin, 3.6x to beta-cyfluthrin and 3.3x to cypermethrin, but was susceptible to beta-cypermethrin and lambda-cyhalothrin. Rhodnius prolixus CA strain showed resistance to all the pyrethroids evaluated, the resistance ratios ranging between 4.5x to lambda-cyhalothrin and 12.4x to cypermethrin. Deltamethrin resistance in both strains was decreased by piperonyl butoxide, suggesting oxidative metabolism as cause of resistance.