Monitoring Rhodnius neglectus (Lent, 1954) populations' susceptibility to insecticide used in controlling actions in urban areas northwest of São Paulo state.

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.

Evaluation of the insecticide susceptibility profile in Cimex lectularius (Hemiptera: Cimicidae) in Belo Horizonte (Brazil).

INTRODUCTION: Since 2013, major Cimex lectularius infestations have been detected in public shelters in Belo Horizonte (Brazil). Due to this, insecticide resistance has been investigated as one of the possible causes for the failure to control bedbugs. METHODS: Cimicids were subjected to bioassays according to the World Health Organization recommendations using deltamethrin and all commercial insecticides available for control of Cimex in Brazil. RESULTS: Cimicids were deltamethrin resistant and presented indicative of resistance to other insecticides, except for propoxur 1%. CONCLUSIONS: The commercial insecticides have a limited effect on bedbug populations, which may justify the unsatisfactory control observed in the shelters studied.

Bedbug salivation patterns during hematophagy in the skin of a mammalian host.

Cimex lectularius (Hemiptera:Cimicidae) infestations have increased over the past decades in several parts of the world, constituting a major urban pest with no reversion signs. The impact on human health caused by these insects, commonly known as bedbugs, is associated with their obligatory hematophagous habit. Allergies induced by hematophagous arthropod bites are related to the deposition of salivary molecules in the host tissues. Many reports of humans developing severe allergic reactions due to bedbug bites have been recorded, however, there is limited information on the salivation of bedbugs on the host, which was the objective of this study. C. lectularius females were fed on blood containing acridine orange fluorochrome, which labeled the principal salivary glands content. The salivation pattern of bedbugs was investigated using intravital microscopy during its blood meal on the ear skin of hairless mice. Saliva deposition occurred during all insect blood-feeding phases, beginning as soon as the mouthpart touched the host skin. During the probing phase, saliva was deposited in large quantities in the host dermis. In contrast, during the engorgement phase (which represents the largest blood meal of the insects), saliva was released at a much slower rate. The apparent release of saliva into the cannulated vessel and/or adjacent tissue occurs only sporadically during insect blood ingestion. However, a small area (spot) of fluorescence was detected around the proboscis tip during this feeding phase. An interesting feature of bedbugs is that they release saliva inside and outside the vessels without removing their mouthparts from the vessel lumen. This is an effective feeding strategy because it does not interrupt blood ingestion and decreases the mouthparts movements on the host's skin, minimizing the damage to tissues and contact time with the host (feeding time).

Susceptibility of phlebotomine sandflies (Diptera: Psychodidae) collected in the field, to alpha-cypermethrin in four municipalities endemic to leishmaniasis.

Although the chemical control against leishmaniasis began in 1953 in Brazil, little information is available on how this strategy has affected populations of phlebotomine sandflies in the field. The objective of this study was to analyze the susceptibility profile of four populations of phlebotomine sandflies to the insecticide alpha-cypermethrin. Sandflies collected in field in four Brazilian municipalities were evaluated using CDC bottles in different concentrations of alpha-cypermethrin. A total of 1,186 phlebotomine sandflies were used in the bioassays. The LD50 ranged from 1.48 to 2.57 ug/mL in the field populations. For a dose of 5 ug/mL of alpha-cypermethrin, the LT50 and LT95 ranged from 17.9 to 27.5 minutes, and LT95 from 39.7 to 61.5 minutes, respectively. All the populations studied were highly susceptible to the insecticide alpha-cypermethrin. Routine studies are needed to detect changes in sandflies susceptibility to insecticides.

The role of LuloPAT amino acid/proton symporters in midgut alkalinization in the sandfly Lutzomyia longipalpis (Diptera - Psychodidae).

In Lutzomyia longipalpis females, which are the main vectors of Leishmania infantum in the Americas, hematophagy is crucial for ovary development. The control of pH in the midgut during blood digestion is important to the functioning of the digestive enzymes, which release amino acids in the luminal compartment that are then transported through the enterocytes to the hemolymph for delivery to the ovary and other organs. In the present work, we investigated transport systems known as LuloPATs that are present in the midgut of L. longipalpis but not in other organs. These transporters achieve symport of amino acids with H+ ions, and one of them (LuloPAT1) is orthologous to a transporter described in Aedes aegypti. According to our results, the transcription levels of LuloPAT1 increased significantly immediately after a blood meal. Based on the variation of the fluorescence of fluorescein with the pH of the medium, we developed a technique that shows the acidification of the cytoplasm of gut cells when amino acids are cotransported with H+ from the lumen into the enterocytes. In our experiments, the midguts of the sandflies were dissected and opened longitudinally so that added amino acids could enter the enterocytes via the lumen (PAT carriers are apical). LuloPAT1 transporters are part of a complex of mechanisms that act synergistically to promote gut alkalinization as soon as blood intake by the vector occurs. In dissected but not longitudinally opened midguts, added amino acids could only enter through the basolateral region of enterocytes. However, alkalinization of the lumen was observed because the entry of some amino acids into the cytoplasm of enterocytes triggers a luminal alkalinization mechanism independent of LuloPATs. These findings provide new perspectives that will enable the characterization of the set of signaling pathways involved in pH regulation within the L. longipalpis midgut.

Resistance to deltamethrin by domestic and wild Triatoma infestans populations in the municipality of Toro Toro, Potosi, Bolivia.

BACKGROUND: Chemical control with pyrethroid insecticides has been effective in reducing endemic areas of distribution of Triatoma infestans in the Southern Cone, as well as Bolivia; this had considerably reduced the infestation of households in a large part of the territory. Nowadays, areas such as the Chaco and the Inter-Andean Valleys are regions where the reach of vector control strategies is becoming limited, and infestations of insecticide-treated households are reported more often. The objective of this study was to determine if the persistence of T. infestans stems from changes in the susceptibility of its toxicological profile in four communities in the municipality of Toro Toro, Potosi, Bolivia. METHODS: Susceptibility to deltamethrin of wild and domestic populations of T. infestans was evaluated in two stages (16 populations before and 13 populations after spraying) among DUs (structures in the intra- and peridomicile) and wild ecotopes, in four communities. Serial dilutions of deltamethrin in acetone (0.2 µl) were applied topically on standardized first-stage nymphs. Dose-response results were analyzed with the software PoloPlus and the relationships between lethal doses (LD) and resistance ratios (RR50) were determined. RESULTS: Different degrees of RR50 were detected among the populations before and after spraying (25.66-54.70 and 21.91-40.67, respectively), as well as in different ecotopes within a DU (DU JC 3, 28.06-36.13, in mixed structures of corrals and chicken coops; and DU JG 3, 46.27-25.70, in kitchen roofs), or in the wild environment of the community JG Sil (29.21-40.67). The mortality of insects undergoing diagnostic dose (DD) was never higher than 34%. CONCLUSION: The results obtained in this study showed resistance of T. infestans to deltamethrin in four communities, hence the complexity of this phenomenon is not only limited to the level of communities, but also applies to the microgeographical level, as in different ecotopes present within the DUs. This phenomenon should be considered while planning the activities of control programs.

Genetic characterization of residual Triatoma infestans populations from Brazil by microsatellite.

In spite of long-term efforts to eliminate Triatoma infestans (Klug 1834) from Brazil, residual foci still persist in the states of Bahia and Rio Grande do Sul. Data on the genetic variability and structuring of these populations are however lacking. Using nine microsatellite loci, we characterized one residual T. infestans population from Bahia and four from Rio Grande do Sul, and compared them with bugs originally from an older focus in São Paulo; 224 bugs were genotyped. The number of alleles per locus ranged from 5 to 11. Observed and expected heterozygosities per locus ranged, respectively, from 0 to 0.786 and from 0 to 0.764. Significant departures from Hardy-Weinberg equilibrium, mainly due to heterozygote deficits, were detected in all loci and in most populations. Global indices estimated by AMOVA were: Fis was 0.37; Fst was 0.28; and Fit was 0.55; overall indices with p = 0.00 indicated substantial differentiation. Inter-population Fst ranged from 0.118 to 0.562, suggesting strong genetic structuring and little to no gene flow among populations. Intra-population Fis ranged from 0.301 to 0.307. Inbreeding was apparent in all populations except that from Bahia-which might be either linked by gene flow to nearby unsampled populations or part of a relatively large local population. The overall pattern of strong genetic structuring among pyrethroid-susceptible residual T. infestans populations suggests that their persistence is probably due to operational control failures. Detection and elimination of such residual foci is technically feasible and must become a public health priority in Brazil.

Risk of ingesting As, Cd, and Pb in animal products in north Rio de Janeiro state, Brazil.

This study evaluated the levels of As, Cd, and Pb in muscle and liver the cattle and chicken. The risk was estimated for the adult population of a midsized city in southeast Brazil, concerning the tolerable ingestion and cancer risk. Samples of muscle and liver (cattle and chicken) were collected (n = 250). Samples of mineral supplements for cattle (n = 4) and chicken feed samples (n = 4) were evaluated as one of many potential source of contamination. Muscle, liver, mineral supplement, and feed samples were dissolved in acid medium and analyzed by ICP-OES. Daily muscle and liver intake was estimated using a questionnaire (N = 427). Daily intake of trace elements by the population based on the consumption of cattle muscle, cattle liver, chicken muscle, and chicken liver was low, corresponding to 2.76%, 0.33%, 2.12%, and 0.22% of the Tolerable Intake defined by the WHO for As; 0.54%, 0.29% 0.55%, 0.01%, for Cd; and 0.80%, 0.07%, 0.62%, 0.02%, for Pb. The mean of total ingestion of As, Cd and Pb was 5.43%, 1.18% and 1.51%, respectively of Tolerable Intake defined by WHO. Cancer risk was lower than 5 × 10-5 year-1. The results indicate that the muscle and liver consumption is a source of As, Cd, and Pb. Consumers that ingest cattle and chicken muscle need attention in terms the risk of cancer related to intake of As and Cd. Feed and mineral supplementation remain as one of many sources of exposure of As, Cd, and Pb.

Assessing the mitochondrial DNA diversity of the Chagas disease vector Triatoma sordida (Hemiptera: Reduviidae).

Triatoma sordida is a species that transmits Trypanosoma cruzi to humans. In Brazil, T. sordida currently deserves special attention because of its wide distribution, tendency to invade domestic environments and vectorial competence. For the planning and execution of control protocols to be effective against Triatominae, they must consider its population structure. In this context, this study aimed to characterise the genetic variability of T. sordida populations collected in areas with persistent infestations from Minas Gerais, Brazil. Levels of genetic variation and population structure were determined in peridomestic T. sordida by sequencing a polymorphic region of the mitochondrial cytochrome b gene. Low nucleotide and haplotype diversity were observed for all 14 sampled areas; π values ranged from 0.002-0.006. Most obtained haplotypes occurred at low frequencies, and some were exclusive to only one of the studied populations. Interpopulation genetic diversity analysis revealed strong genetic structuring. Furthermore, the genetic variability of Brazilian populations is small compared to that of Argentinean and Bolivian specimens. The possible factors related to the reduced genetic variability and strong genetic structuring obtained for studied populations are discussed in this paper.

Assessing the mitochondrial DNA diversity of the Chagas disease vector Triatoma sordida (Hemiptera: Reduviidae)

Triatoma sordida is a species that transmits Trypanosoma cruzi to humans. In Brazil, T. sordida currently deserves special attention because of its wide distribution, tendency to invade domestic environments and vectorial competence. For the planning and execution of control protocols to be effective against Triatominae, they must consider its population structure. In this context, this study aimed to characterise the genetic variability of T. sordida populations collected in areas with persistent infestations from Minas Gerais, Brazil. Levels of genetic variation and population structure were determined in peridomestic T. sordida by sequencing a polymorphic region of the mitochondrial cytochrome b gene. Low nucleotide and haplotype diversity were observed for all 14 sampled areas; π values ranged from 0.002-0.006. Most obtained haplotypes occurred at low frequencies, and some were exclusive to only one of the studied populations. Interpopulation genetic diversity analysis revealed strong genetic structuring. Furthermore, the genetic variability of Brazilian populations is small compared to that of Argentinean and Bolivian specimens. The possible factors related to the reduced genetic variability and strong genetic structuring obtained for studied populations are discussed in this paper.

Markers for the population genetics studies of Triatoma sordida (Hemiptera: Reduviidae).

BACKGROUND: Triatoma sordida, a vector of Trypanosoma cruzi, is native of Brazil, Bolivia, Paraguay, Argentina, and Uruguay, and occurs primarily in peridomiciles. Currently, it is the species most frequently captured by the Chagas Disease Control Program in Brazil. For this reason, population genetic studies attract great interest, as they can provide further information about the dispersal and household invasion processes of this species. In the absence of suitable markers, the objective of this study was to test the cross amplification of microsatellite primers. FINDINGS: 23 primers were tested for microsatellite loci already described for other species of the genus Triatoma sp. Forty four specimens of T. sordida captured in the north of Minas Gerais were used to validate the use of standardized loci for population genetic analyses. It was possible to amplify 10 of the 23 loci tested for T. sordida. CONCLUSIONS: This is the first study that provides 10 microsatellite markers for population analysis of this triatomine species. Cross-amplification of primers can be used among other phylogenetically related species whose loci are already available for study.

Susceptibility characterization of residual Brazilian populations of Triatoma infestans Klug, 1834 (Hemiptera: Reduviidae) to deltamethrin pyrethroid.

INTRODUCTION: Despite years of efforts towards the elimination of Triatoma infestans in Brazil, residual foci still persist in some areas of the States of Bahia and Rio Grande do Sul. The persistence of these T. infestans populations in the country has two different origins of equal concern: operational failures or insecticide resistance. Thus, the objective of this study was to characterize the susceptibility profile of the residual Brazilian populations of T. infestans to deltamethrin. METHODS: The susceptibility reference lineage was derived from Cipein/Argentina. The populations studied were manually collected using a dislodging agent in peridomiciles in the States of Bahia (Novo Horizonte) and of Rio Grande do Sul (Santa Rosa and Doutor Maurício Cardoso). Serial dilutions of deltamethrin were prepared and applied at the dorsal abdomen of first instar nymphs. The control group received only pure acetone. Mortality was evaluated after 72h. Qualitative tests assessed the mortality of a diagnostic dose of 1xLD99 (2.76ng a.i./nymph) determined for the susceptibility reference lineage. RESULTS: The susceptibility profile characterization of the T. infestans populations revealed an RR50 ranging from 1.73 to 3.26. The mortality percentage in response to a diagnostic dose was 100%. The results obtained in the quantitative and qualitative assays corresponded for all populations. CONCLUSIONS: The results of this study indicate that the persistence of residual foci of T. infestans in Bahia and Rio Grande do Sul is not related to insecticide resistance but may be associated with operational failures. In Rio Grande do Sul, we must consider the possibility of continuous reinfestation by Argentinian individuals, which justifies active and efficient epidemiological surveillance.

Toxicological profile of deltamethrin in Triatoma brasiliensis (Hemiptera: Reduviidae) in State of Ceará, Northeastern Brazil.

INTRODUCTION: Triatoma brasiliensis is the species of greatest epidemiological relevance in the semi-arid region of Brazil. This species is predominantly found in domestic environments, and it has the ability to build large colonies with high levels of natural infection via Trypanosoma cruzi. Thus, T. brasiliensis is one of the most efficient transmitters of Chagas disease (CD) to humans. Despite household spraying with residual insecticides, many areas report persistent reinfestations for reasons that remain poorly understood. Therefore, this study sought to characterize the toxicological profile of deltamethrin in T. brasiliensis from areas with persistent reinfestation in State of Ceará, Brazil. METHODS: The susceptibility reference lineage (SRL) was derived from Umari. Serial dilutions of deltamethrin were prepared and applied to the dorsal abdomen of first instar nymphs. The control group received only pure acetone. Mortality was evaluated after 72h. Qualitative tests assessed mortality in response to a diagnostic dose of 1xLD99 (0.851 nanograms of active ingredient per treated nymph) of the SRL. RESULTS: The susceptibility profile characterization of the T. brasiliensis populations revealed 50% resistance ratios (RR50) that ranged from 0.32 to 1.21. The percentage of mortality in response to the diagnostic dose was 100%. CONCLUSIONS: We demonstrated that T. brasiliensis was highly susceptible to deltamethrin. The control difficulties found might be related to the recolonization of the triatomines originating from neighboring environments and the possible operational failures related to the process of spraying that enabled specimens less susceptible to deltamethrin to survive.

Standardization of laboratory bioassays for the study of Triatoma sordida susceptibility to pyrethroid insecticides.

BACKGROUND: Increasing reports of high-resistant Triatominae populations concerns scientists and sanitarians as little is known about the factors behind the occurrence of such phenotype and its real impact on vector control strategies. Moreover, the utilization of a large variety of methodologies hinder the comparison of the reported studies. METHODS: This work aims to review laboratory bioassays, redefining the assessed biological features (age, generation and insecticide application area) and technical procedures (mortality recording time and the ideal diagnostic dose). RESULTS: Results were not influenced by the insecticide application area in nymphs or by their generation. Three days-old specimen's revealed lower susceptibility to the tested insecticide. We determined that it is more appropriate to record mortality 72 h after treatment with insecticide, as well as using a diagnostic dose of 1xDL99. CONCLUSION: This work suggests more adequate methodological parameters for assessing insecticide resistance in triatomines, which also allows the comparison of results obtained by different research groups. For laboratory bioassays, we recommend: 1) the use of first instar nymphs from first or second generation; 2) 3 day-old specimens; 2) application of insecticide in the dorsal or ventral abdomen area; 3) mortality recording 72 h after treatment with pyrethroids and 4) a diagnostic dose of 1x LD99.

Susceptibility characterization of residual Brazilian populations of Triatoma infestans Klug, 1834 (Hemiptera: Reduviidae) to deltamethrin pyrethroid

INTRODUCTION: Despite years of efforts towards the elimination of Triatoma infestans in Brazil, residual foci still persist in some areas of the States of Bahia and Rio Grande do Sul. The persistence of these T. infestans populations in the country has two different origins of equal concern: operational failures or insecticide resistance. Thus, the objective of this study was to characterize the susceptibility profile of the residual Brazilian populations of T. infestans to deltamethrin. METHODS: The susceptibility reference lineage was derived from Cipein/Argentina. The populations studied were manually collected using a dislodging agent in peridomiciles in the States of Bahia (Novo Horizonte) and of Rio Grande do Sul (Santa Rosa and Doutor Maurício Cardoso). Serial dilutions of deltamethrin were prepared and applied at the dorsal abdomen of first instar nymphs. The control group received only pure acetone. Mortality was evaluated after 72h. Qualitative tests assessed the mortality of a diagnostic dose of 1xLD99 (2.76ng a.i./nymph) determined for the susceptibility reference lineage. RESULTS: The susceptibility profile characterization of the T. infestans populations revealed an RR50 ranging from 1.73 to 3.26. The mortality percentage in response to a diagnostic dose was 100%. The results obtained in the quantitative and qualitative assays corresponded for all populations. CONCLUSIONS: The results of this study indicate that the persistence of residual foci of T. infestans in Bahia and Rio Grande do Sul is not related to insecticide resistance but may be associated with operational failures. In Rio Grande do Sul, we must consider the possibility of continuous reinfestation by Argentinian individuals, which justifies active and efficient epidemiological surveillance. .

Toxicological profile of deltamethrin in Triatoma brasiliensis (Hemiptera: Reduviidae) in State of Ceará, Northeastern Brazil

INTRODUCTION: Triatoma brasiliensis is the species of greatest epidemiological relevance in the semi-arid region of Brazil. This species is predominantly found in domestic environments, and it has the ability to build large colonies with high levels of natural infection via Trypanosoma cruzi. Thus, T. brasiliensis is one of the most efficient transmitters of Chagas disease (CD) to humans. Despite household spraying with residual insecticides, many areas report persistent reinfestations for reasons that remain poorly understood. Therefore, this study sought to characterize the toxicological profile of deltamethrin in T. brasiliensis from areas with persistent reinfestation in State of Ceará, Brazil. METHODS: The susceptibility reference lineage (SRL) was derived from Umari. Serial dilutions of deltamethrin were prepared and applied to the dorsal abdomen of first instar nymphs. The control group received only pure acetone. Mortality was evaluated after 72h. Qualitative tests assessed mortality in response to a diagnostic dose of 1xLD99 (0.851 nanograms of active ingredient per treated nymph) of the SRL. RESULTS: The susceptibility profile characterization of the T. brasiliensis populations revealed 50% resistance ratios (RR50) that ranged from 0.32 to 1.21. The percentage of mortality in response to the diagnostic dose was 100%. CONCLUSIONS: We demonstrated that T. brasiliensis was highly susceptible to deltamethrin. The control difficulties found might be related to the recolonization of the triatomines originating from neighboring environments and the possible operational failures related to the process of spraying that enabled specimens less susceptible to deltamethrin to survive. .

Effects of preheating and precooling on the hardness and shrinkage of a composite resin cured with QTH and LED.

The aim of this study was to evaluate in vitro the hardness and shrinkage of a pre-cooled or preheated hybrid composite resin cured by a quartz-tungsten-halogen light (QTH) and light-emitting diode (LED) curing units. The temperature on the tip of the devices was also investigated. Specimens of Charisma resin composite were produced with a metal mold kept under 37°C. The syringes were submitted to 4°C, 23°C, and 60°C (n=20) before light-curing, which was carried out with the Optilux 501 VCL and Elipar FreeLight 2 units for 20 seconds. The specimens were kept under 37°C in a high humidity condition and darkness for 48 hours. The Knoop hardness test was carried out with a 50 gram-force (gf) load for 10 seconds, and the measurement of the shrinkage gap was carried out using an optical microscope. The data were subjected to analysis of variance and the Games-Howell test (α=0.05). The mean hardness of the groups were similar, irrespective of the temperatures (p>0.05). For 4°C and 60°C, the top surface light-cured by LED presented significantly reduced shrinkage when compared with the bottom and to both surfaces cured by QTH (p<0.05). It was concluded that the hardness was not affected by pre-cooling or preheating. However, polymerization shrinkage was slightly affected by different pre-polymerization temperatures. The QTH-curing generated greater shrinkage than LED-curing only when the composite was preheated. Different temperatures did not affect the composite hardness and shrinkage when cured by a LED curing unit.

The influence of surface standardization of lithium disilicate glass ceramic on bond strength to a dual resin cement.

In vitro studies to assess bond strength between resins and ceramics have used surfaces that have been ground flat to ensure standardization; however, in patients, ceramic surfaces are irregular. The effect of a polished and unpolished ceramic on bond strength needs to be investigated. Sixty ceramic specimens (20×5×2 mm) were made and divided into two groups. One group was ground with 220- to 2000-grit wet silicon carbide paper and polished with 3-, 1-, and »-µm diamond paste; the other group was neither ground nor polished. Each group was divided into three subgroups: treated polished controls (PC) and untreated unpolished controls (UPC), polished (PE) and unpolished specimens (UPE) etched with hydrofluoric acid, and polished (PS) and unpolished specimens (UPS) sandblasted with alumina. Resin cement cylinders were built over each specimen. Shear bond strength was measured, and the fractured site was analyzed. Analysis of variance (ANOVA) and Tukey post hoc tests were performed. PE (44.47 ± 5.91 MPa) and UPE (39.70 ± 5.46 MPa) had the highest mean bond strength. PS (31.05 ± 8.81 MPa), UPC (29.11 ± 8.11 MPa), and UPS (26.41 ± 7.31 MPa) were statistically similar, and PC (24.96 ± 8.17 MPa) was the lowest. Hydrofluoric acid provides the highest bond strength regardless of whether the surface is polished or not.

Real-time PCR investigation on the expression of sboA and ituD genes in Bacillus spp.

AIMS: To investigate the expression of sboA and ituD genes among strains of Bacillus spp. at different pH and temperature. METHODS AND RESULTS: Different Bacillus strains from the Amazon basin and Bacillus subtilis ATCC 19659 were investigated for the production of subtilosin A and iturin A by qRT-PCR, analysing sboA and ituD gene expression under different culture conditions. Amazonian strains presented a general gene expression level lower than B. subtilis ATCC 19659 for sboA. In contrast, when analysing the expression of ituD gene, the strains from the Amazon, particularly P40 and P45B, exhibited higher levels of expression. Changes in pH (6 and 8) and temperature (37 and 42 °C) caused a decrease in sboA expression, but increased ituD expression among strains from Amazonian environment. CONCLUSIONS: Temperature and pH have an important influence on the expression of genes sboA (subtilosin A) and ituD (iturin A) among Bacillus spp. The strains P40 and P45B can be useful for the production of antimicrobial peptide iturin A. SIGNIFICANCE AND IMPACT OF THE STUDY: Monitoring the expression of essential biosynthetic genes by qRT-PCR is a valuable tool for optimization of the production of antimicrobial peptides.

Susceptibility of triatoma infestans to deltamethrin in Rio Grande do Sul, Brazil.

Strategies for controlling Chagas disease are based on spraying infested houses with pyrethroid insecticides. However, the intense use of these insecticides has promoted resistance of Triatoma infestans and, in Argentina, Bolivia and Southern Brazil, low levels of resistance have been reported. Due to the persistence of T. infestans in the state of Rio Grande do Sul (RS), we evaluated the occurrence of deltamethrin resistance in four strains from different municipalities in comparison to two susceptible strains from Brazil and one resistant strain from Bolivia. The results indicated the absence of resistance in T. infestansfrom RS.