Thuringiensis Israelensis Research Articles

Malaria mosquito control in rice paddy farms using biolarvicide mixed with fertilizer in Tanzania: semi-field experiments

BackgroundThe wide distribution of malaria mosquito breeding sites within tropical environments limits the mosquito larval source management efforts to control malaria. Rice farming contributes substantially in supporting malaria mosquito productivity in tropical countries. To overcome this challenge, this study was carried out to determine the effect of applying a mixture of biolarvicide and fertilizer on mosquito larvae density in rice farms under semi-field conditions in Tanzania.MethodsA semi-field experiment was designed to determine the timing of application of a biolarvicide, Bacillus thuringiensis israelensis (Bti) and fertilizer (di-ammonium phosphate-DAP or urea) and assess their effect on mosquito larvae density and rice grain outputs. The experiment had five blocks (4 treatment arms and one control arm) and each had four replicates. Treatment arms had different intervals of days between treatments for mixtures of fertilizer and biolarvicides. The dosages used were 10 g of Bti/16 M2 and 160 g of DAP/Urea/16 m2.ResultsIn overall, the intervention blocks (with biolarvicide) had lowest mean mosquito larvae abundance compared to control block (F = 22.42, P < 0.001). Similarly, the control arm maintained highest density of Anopheles gambiae sensu lato larvae compared to interventions blocks (F = 21.6, P < 0.001). The best determined timing for application of Bti was in 7 and in 10 days (F = 3.753, P < 0.001). There was neither significant different in mean rice grain harvest per ten panicle (F = 1.453, P = 0.27) nor mean difference in rice grain harvest (F = 1.479, P = 0.26) per intervention arms.ConclusionThe findings of this study have shown that application of a mixture of Bti and fertilizer have impact on both mosquito larvae density and maintaining yield rice harvest. Thus, application of a combination of biolarvicide and fertilizer can be an alternative approach in malaria mosquito intervention among rice farming communities of rural Tanzania.

Cloning, expression and activity of ATP-binding protein in Bacillus thuringiensis toxicity modulation against Aedes aegypti

BackgroundBacillus thuringiensis israelensis (Bti) is a widely used mosquitocidal microbial pesticide due to its high toxicity. ATP-binding proteins (ABP) are prevalently detected in insects and are related to reaction against Bti toxins. However, the function of ABP in mosquito biocontrol is little known, especially in Aedes aegypti. Therefore, this study aimed to clarify the function of ABP in Ae. aegypti against Bti toxin.ResultsAedes aegypti ABP (GenBank: XM_001661856.2) was cloned, expressed and purified in this study. Far-western blotting and ELISA were also carried out to confirm the interaction between ABP and Cry11Aa. A bioassay of Cry11Aa was performed both in the presence and absence of ABP, which showed that the mortality of Ae. aegypti is increased with an increase in ABP.ConclusionsOur results suggest that ABP in Ae. aegypti can modulate the toxicity of Cry11Aa toxin to mosquitoes by binding to Bti toxin. This could not only enrich the mechanism of Bt toxin, but also provide more data for the biocontrol of this transmission vector.

Mosquito control based on Bacillus thuringiensis israelensis (Bti) interrupts artificial wetland food chains

The biocide Bacillus thuringiensis israelensis (Bti) has become the most commonly used larvicide to control mosquitoes in seasonal wetlands. Although Bti is considered non-toxic to most aquatic organisms, the non-biting chironomids show high susceptibilities towards Bti. As chironomids are a key element in wetland food webs, major declines in their abundance could lead to indirect effects that may be passed through aquatic and terrestrial food chains. We conducted two mesocosm experiments to address this hypothesis by assessing direct and indirect effects of Bti-modified availability of macroinvertebrate and zooplankton food resources on the predatory larvae of palmate and smooth newts (Urodelans: Lissotriton helveticus, Lissotriton vulgaris). We examined newt survival rates and dietary composition by means of stable isotope (δ15N and δ13C) analysis in the presence of Bti treatment and a predator (Odonata: Aeshna cyanea). We assessed palmate newts' body size at and time to metamorphosis while developing in Bti treated mesocosms. Chironomid larvae were the most severely affected aquatic invertebrates in all Bti treated food chains and experienced abundance reductions by 50 to 87%. Moreover, stable isotope analysis revealed that chironomids were preferred over other invertebrates and comprised the major part in newts' diet (56%) regardless of their availability. The dragonfly A. cyanea decreased survival of newt larvae by 27% in Bti treated mesocosms showing affected chironomid abundances. Increasing intraguild predation is most likely favored by the Bti-induced reduction of alternative prey such as chironomid larvae. The decreased food availability after Bti treatment led to slightly smaller L. helveticus metamorphs while their developmental time was not affected. Our findings highlight the crucial role of chironomids in the food webs of freshwater ecosystems. We are also emphasizing the importance of reconsidering human-induced indirect effects of mosquito control on valuable wetland ecosystems particularly in the context of worldwide amphibian and insect declines.

2018 Highlights of Mosquito and Vector Biology and Control in Latin America.

The 28th Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 84th Annual Meeting of the AMCA held in Kansas City, MO, in February 2018. The Latin American Symposium promotes the participation of vector control specialists, public health workers, and academic members from Latin America and the sharing of scientific data between continents. Generally, presentations are in Spanish with simultaneous translation into English. The majority of presentation slides are in English to facilitate communication among all meeting attendees. This publication includes summaries of 14 oral presentations by participants from Colombia, Mexico, and the United States of America. Topics addressed in the 3 sessions of the symposium included: larval habitats, insecticide resistance, new Aedes mosquito traps, blood meal analysis and fitness outcomes, vertical transmission of dengue, and transstadial composition of midgut microbiota. Control techniques discussed included sterile insect technique (SIT) with radiation, SIT by Wolbachia, thermal fogging, ultra-low volume pesticide applications, indoor residual spraying, Bacillus thuringiensis israelensis de Barjac (Bti), and Spinosad larval treatments. Presentations were also given on species composition and diversity of phlebotomine sand flies. Presentations were related to the vector species belonging to the mosquito genera, Anopheles, Aedes, and Culex as well as phlebotomine sand flies involved in the transmission of the causal agents of malaria, arboviruses (dengue, chikungunya, Zika), and leishmaniasis.

Reduction of Malaria Vector Mosquitoes in a Large Scale Intervention Trial in Rural Burkina Faso Using &lt;i&gt;Bti&lt;/i&gt; Based Larval Source Management

Background: Malaria remains one of the most important causes of morbidity and death in Sub-Saharan Africa. Along with early diagnosis and treatment of malaria cases and intermittent preventive treatment in pregnancy (IPTp), vector control is an important tool in the reduction of new cases. Alongside the use of long lasting insecticide treated bed nets (LLINs) and indoor residual spraying (IRS), targeting the vector larvae with biological larvicides such as Bacillus thuringiensis israelensis (Bti) is gaining importance as a means of reducing the number of mosquito larvae before they emerge to their adult stage. With this study, we present data corroborating the entomological impact of such an intervention in a rural African environment. Methods: The study extended over two years and researched the impact of biological larviciding with Bti on malaria mosquitoes that were caught indoors and outdoors of houses using light traps. We calculated the achieved reductions in female Anopheles mosquitoes for two different larviciding choices using a regression model. Findings: In villages that received selective treatment of the most productive breeding sites, the number of female Anopheles spp. dropped by 61% (95% CI: 54%-66%) compared to the pre-intervention period. In villages in which all breeding sites were treated, the number of female Anopheles spp. was reduced by 70% (95% CI: 64%-74%) compared to the pre-intervention period. Interpretation: We showed that malaria vector abundance can be dramatically reduced through larviciding of breeding habitats and that, in many geographical settings, they are a viable addition to current malaria control measures. Trial Registration Information: The study was registered under the trial id PACTR201611001721299 on the Pan African Clinical Trials Registry.. Funding Statement: This work was financed by Manfred Lautenschlager foundation, Wiesloch, Germany. Declaration of Interests: The authors declare no competing interests. Ethics Approval Statement: The study was approved by the ethics committees of the University of Heidelberg under the certificate number S-438/2013 and additionally presented to and granted by the national ethics board of Burkina Faso in Ouagadougou and the local ethics committee at the research site in Nouna. We collected aggregated collective informed consent for the spraying activities for each village.

Detection of spoilage associated bacteria using Raman-microspectroscopy combined with multivariate statistical analysis

Raman-Microspectroscopy with subsequent chemometric evaluation was used for the rapid and non-destructive differentiation of seven important spoilage related microorganisms, namely Brochothrix thermosphacta DSM 20171, Pseudomonas fluorescens DSM 4358, Pseudomonas fluorescens DSM 50090, Micrococcus luteus, Escherichia coli HB101, Escherichia coli TOP10 and Bacillus thuringiensis israelensis DSM 5724. Therefore fast collected spectra directly from rapid surface blots without any pretreatments like purification or singulation steps were used. To estimate and classify the Raman-spectroscopic data at genera and strain level an adequate preprocessing together with a subsequent chemometric evaluation consisting of principal component analysis and discriminant analysis was used. Thereby, importance was attached to a balanced data set, as this makes the multivariate analysis of the data significantly more resilient and meaningful. The analysis showed that the differentiation of spoilage related microorganisms on genera and strain level was successful and the classification of independent test data showed only an error rate of 3.5%.

Alternative insecticides for larval control of the dengue vector Aedes aegypti in Lao PDR: insecticide resistance and semi-field trial study

BackgroundThe mosquito Aedes aegypti is the primary vector of several arboviruses, such as dengue, chikungunya and Zika, and represents a major public health problem in Southeast Asia. In Laos, where dengue is reemerging, several Ae. aegypti populations from the capital Vientiane have shown resistance to the organophosphate temephos, a commonly-used larvicide for public health interventions.MethodsHere, we tested the insecticide susceptibility of a wild larval population of Ae. aegypti against Bacillus thuringiensis israelensis (Bti), diflubenzuron, pyriproxyfen and spinosad. Residual efficacies of Bti (VectobacWG®), diflubenzuron (Killmos®) and temephos (Abate®) were then evaluated under simulated field conditions against the wild Ae. aegypti population.ResultsThe larval bioassays showed that the wild Ae. aegypti strain was moderately resistant to temephos and spinosad (resistance ratio, RR < 5) and fully susceptible to the other insecticides (RR = 1). The simulated field trial bioassays showed that all of the insecticides tested remained above the WHO acceptable larvicide threshold after 28 weeks.ConclusionsThese results suggest that Bti and diflubenzuron may be promising alternative larvicides for controlling dengue vectors in water-storage containers in Laos, especially against Ae. aegypti populations, in which resistance to temephos has been detected.

Adverse effects of mosquito control using Bacillus thuringiensis var. israelensis: Reduced chironomid abundances in mesocosm, semi-field and field studies

The ecological consequences of mosquito control using the larvicide Bacillus thuringiensis israelensis (Bti) are still a matter of debate especially when it comes to adverse effects on non-target but susceptible non-biting midges (Chironomidae). Our work aimed to assess the effects of Bti applied in operational mosquito control rates on chironomid abundances in seasonal flooded freshwater wetlands. We assessed the invertebrate community alongside with aquatic insect emergence rates in studies with increasing ecotoxicological complexity, ranging from artificial mesocosms, over a semi-field approach using enclosures to natural conditions in field studies. Study sites represented different habitats (forest, meadow, floodplain) with regularly Bti treated and untreated temporal wetlands that are located in the Upper Rhine Valley, Germany. Larval chironomids were the most affected group of organism in the altered aquatic invertebrate communities of the Bti treated pond mesocosms. The larvae of the chironomid subfamilies Chironominae and Orthocladiinae were affected by Bti field concentrations while emergence rates of Tanypodinae did not change. The Bti treatment significantly reduced the targeted mosquitoes in the mesocosm and in the field studies. At the same time, however, the application of operational Bti field rates reduced overall chironomid emergence rates to about half of the control rates. The observed abundance reductions occurred independently of the ecological complexity in the study design in artificial mesocosms as well as realistic field conditions. The considerable reduction of the abundant non-target chironomids along with mosquitoes may subsequently lead to unwanted indirect negative effects for birds, bats and other aquatic organisms feeding on midges. Hence, large-scale applications of Bti for mosquito control in seasonal wetlands should be considered more carefully. This is of special importance when these wetlands are parts of national parks, nature reserves or Natura 2000 sites that were created for the protection of nature and environmental health.

The combinatory effect of Cyt1Aa flexibility and specificity against dipteran larvae improves the toxicity of Bacillus thuringensis kurstaki toxins

Cyt1A98 is a novel cytolytic protein, from BUPM98 Bacillus thuringiensis strain, characterized by its synergistic activity with B. thuringiensis kurstaki toxins against lepidopteran larvae. In this study, we evidenced that Cyt1A98 improves the toxicity of B. thuringiensis kurstaki toxins against Aedes aegypti larvae. In fact, the strain BNS3pHTcyt1A98 exhibited a larvicidal activity of about 849-fold of that of BNS3pHTBlue against A. aegypti. The molecular and biochemical characterizations, of cyt1A98 gene and its product, were achieved. Cyt1A98 had an LC50 value of about 126.56 mg l−1 against A. aegypti larvae. Compared to Cyt1Aa of B. thuringiensis israelensis, Cyt1A98 amino acid sequence harbours three substitutions of three conserved amino acids among Cyt1Aa family members (Ser42Pro, Pro82Ala, Met188Thr). The Cyt1A98 protein structural analysis evidenced more flexibility than Cyt1Aa. According to the high fluctuation observed for the residue Pro42, the amino acid at position 42 is implicated in the flexibility property of Cyt1Aa especially for the αC and αD helices, involved in the penetration into the cell membrane. The toxicity improvement could be probably due to the higher flexibility combined with the specific affinity toward dipteran larvae. The Cyt1A/B. thuringiensis kurstaki Cry toxins model provides a potential molecular genetic strategy for an efficient bioinsecticide.

Environment friendly management of mosquito: a short review

Despite the large scale use of insecticides, capacity building, municipality, community and metropolis awareness, and preventive measures to counter vector borne diseases which are mounting day-by-day, new tools are now been introducing to prevent the spread of mosquito transmitted diseases. The low efficacy status of chemical pesticides have lead to the interest of researchers in search of fresh and even more practicable vector control methodologies to be applied. In this regards, multiple alternatives have been monitoring to develop control practice measures for the eradication, observation and control of mosquitoes at larval level by the use of a sustainable biological monitoring and control by an ordinary constructive predator, to exercise monitoring and practical control measures over parasites at larval stages in environmental and eco-friendly techniques. In particular, bio-control measures to monitor and control practical practices, context predatory larvivorous fish, dragonfly nymph, frogs, copepods, turtle, Entomopathogenic bacillus, Bacillus sphaericus and Bacillus thuringiensis israelensis are beings tried in different regions of world. The available research on the subject recommends that there exist multiple direct and indirect growth factors that could play a dynamic role in prey and predator’s survival. Species controphic that have an impact on concerned eco-relation reflect significant effect. In addition to this, certain eco-relations represent positive stimuli for the control of vector borne viral diseases. As a bio-control achieving feasible agent for vector monitoring, pointing, management and control predatory larvivorous fish, dragonfly nymph, frogs, copepods, turtle, Entomopathogenic bacillus, Bacillus sphaericus and Bacillus thuringiensis israelensis are not only considered as a liberated intervention for disease vector control of practices and mechanical control cost deterrents as well. Further research has been suggested on the subject so as to find out even more practicable and effective mosquito monitoring and practical control practices.Bangladesh J. Sci. Ind. Res.53(3), 169-178, 2018

Impact of Storage and Handling Temperatures On the Activities of Mosquito Larvicides.

Larvicides based on Bacillus thuringiensis israelensis, Lysinibacilus sphaericus (formerly Bacillus sphaericus), Saccharopolyspora spinosa and insect growth regulators such as methoprene, and a few others are the most commonly used mosquito control products in the United States and elsewhere because of their relative specificity and high effectiveness against target organisms, safety to nontarget species, and compatibility with the environment. Using standard laboratory bioassay, we have evaluated the loss of larvicidal activities of various formulations after being exposed to different storage and handling temperatures. Transportation of products under outdoor conditions for an extended period of time caused more activity loss compared with other conditions. Results are discussed based on their active ingredients as well as physico-chemical properties of the formulations reported. Recommendations are made for proper inventory control, storage, and handling of larvicides to minimize loss of larvicidal activity.

Oviposition of Aedes aegypti Linnaeus, 1762 and Aedes albopictus Skuse, 1894 (Diptera: Culicidae) under laboratory and field conditions using ovitraps associated to different control agents, Manaus, Amazonas, Brazil

The aim of this study was to analyze the effectiveness of different control agents of Aedes aegypti and Aedes albopictus associated with ovitraps under laboratory and field conditions. Five treatments were used: grass infusion+Bacillus thuringiensis israelensis (gI+Bti), grass infusion+Saccharopolyspora spinosa (gI+Ss), grass infusion+Pyriproxyfen (gI+P), distilled water+Toxorhynchites haemorrhoidalis (dW+Th), and grass infusion (gI) (control). The highest mean number of eggs of both species were obtained with grass infusion in the laboratory. Among control agents, the lowest mean of A. aegypti eggs occurred with gI+Ss and the lowest mean of A. albopictus eggs occurred with dW+Th. There was no difference between treatments in A. aegypti (P=0.4320) and A. albopictus (P=0.7179). In the field, the highest mean number of eggs for both species were obtained with gI+Ss, and the lowest values were obtained with gI+P (P=0.0124). The treatments can be applied to both the surveillance and the control, but ovitraps with biological larvicide Bti were more effective and safer considering the number of eggs laid and selectivity of pathogens for mosquitoes.

Community-based malaria control in southern Malawi: a description of experimental interventions of community workshops, house improvement and larval source management

BackgroundIncreased engagement of communities has been emphasized in global plans for malaria control and elimination. Three interventions to reinforce and complement national malaria control recommendations were developed and applied within the context of a broad-based development initiative, targeting a rural population surrounding a wildlife reserve. The interventions, which were part of a 2-year research trial, and assigned to the village level, were implemented through trained local volunteers, or ‘health animators’, who educated the community and facilitated collective action.ResultsCommunity workshops on malaria were designed to increase uptake of national recommendations; a manual was developed, and training of health animators conducted, with educational content and analytical tools for a series of fortnightly community workshops in annual cycles at village level. The roll-back malaria principle of diagnosis, treatment and use of long-lasting insecticidal nets was a central component of the workshops. Structural house improvement to reduce entry of malaria vectors consisted of targeted activities in selected villages to mobilize the community into voluntarily closing the eaves and screening the windows of their houses; the project provided wire mesh for screening. Corrective measures were introduced to respond to field challenges. Committees were established at village level to coordinate the house improvement activities. Larval source management (LSM) in selected villages consisted of two parts: one on removal of standing water bodies by the community at large; and one on larviciding with bacterial insecticide Bacillus thuringiensis israelensis by trained village committees. Community workshops on malaria were implemented as ‘core intervention’ in all villages. House improvement and LSM were implemented in addition to community workshops on malaria in selected villages.ConclusionsThree novel interventions for community mobilization on malaria prevention and control were described. The interventions comprised local organizational structure, education and collective action, and incorporated elements of problem identification, planning and evaluation. These methods could be applicable to other countries and settings.

Optimization of bio-insecticide production by Tunisian Bacillus thuringiensis israelensis and its application in the field

Bacillus thuringiensis israelensis biological larvicides represent a good approach to support vector borne diseases control programs targeting earlier aquatic larval stages to limit their adult mosquito spreading.BUPM98, a Tunisian B. thuringiensis israelensis strain, is highly active against Aedes aegypti larvae. For the production of BUPM98 bioinsecticide, an agricultural by-product medium consisting of gruel and soybean meal has been optimized. Optimization of the medium composition was performed with statistical methodology based on experimental designs. Accordingly, the optimum toxin production was 2375.36 ± 48.52 mg/L at the following conditions: K2HPO4: 60 mM, pH: 7.6 and soybean meal: 32 g/L, which led to an improvement of 30% compared to the starting conditions. The impact of water quality on the effectiveness of BUPM98 delta-endotoxins against Culex pipiens larvae was investigated. Indeed, under field conditions, the direct application of BUPM98 fermentation product in stagnant water pools reduced the immature density of C. pipiens larvae by more than 95% after 48 h of treatment. Prolonged residual action was observed until 14 days. In conclusion, BUPM98, a local strain of B. thuringiensis israelensis, is very promising bioinsecticide for mosquito population management in Tunisia.

Changes in Larval Mosquito Microbiota Reveal Non-target Effects of Insecticide Treatments in Hurricane-Created Habitats.

Ephemeral aquatic habitats and their associated microbial communities (microbiomes) play important roles in the growth and development of numerous aquatic insects, including mosquitoes (Diptera). Biological control agents, such as Bacillus thuringiensis israelensis (Bti) or insect growth regulators (e.g., methoprene), are commonly used to control mosquitoes in these habitats. However, it is unknown how commonly used control compounds affect the mosquito internal microbiome and potentially alter their life history traits. The objectives of this study were threefold: characterize the internal microbiota of Aedes larvae (Culicidae) in ephemeral forested mosquito habitat using high-throughput amplicon based sequencing, assess how mosquito control treatments affect the internal microbial communities of larval mosquitoes, and determine if changes to the microbiome resulted from direct or indirect treatment effects. The larval microbiome varied in community composition and diversity with development stage and treatment, suggesting potential effects of control compounds on insect microbial ecology. While microbial community differences due to Bti treatment were a result of indirect effects on larval development, methoprene had significant impacts on bacterial and algal taxa that could not be explained by indirect treatment effects. These results provide new information on the interactions between pesticide treatments and insect microbial communities.

Bacterial microbiota of Aedes aegypti mosquito larvae is altered by intoxication with Bacillus thuringiensis israelensis

BackgroundInsect microbiota is a dynamic microbial community that can actively participate in defense against pathogens. Bacillus thuringiensis (Bt) is a natural entomopathogen widely used as a bioinsecticide for pest control. Although Bt’s mode of action has been extensively studied, whether the presence of microbiota is mandatory for Bt to effectively kill the insect is still under debate. An association between a higher tolerance and a modified microbiota was already evidenced but a critical point remained to be solved: is the modified microbiota a cause or a consequence of a higher tolerance to Bt?MethodsIn this study we focused on the mosquito species Aedes aegypti, as no work has been performed on Diptera on this topic to date, and on B. thuringiensis israelensis (Bti), which is used worldwide for mosquito control. To avoid using antibiotics to cure bacterial microbiota, mosquito larvae were exposed to an hourly increasing dose of Bti during 25 hours to separate the most susceptible larvae dying quickly from more tolerant individuals, with longer survival.ResultsDenaturing gradient gel electrophoresis (DGGE) fingerprinting revealed that mosquito larval bacterial microbiota was strongly affected by Bti infection after only a few hours of exposure. Bacterial microbiota from the most tolerant larvae showed the lowest diversity but the highest inter-individual differences. The proportion of Bti in the host tissue was reduced in the most tolerant larvae as compared to the most susceptible ones, suggesting an active control of Bti infection by the host.ConclusionsHere we show that a modified microbiota is associated with a higher tolerance of mosquitoes to Bti, but that it is rather a consequence of Bti infection than the cause of the higher tolerance. This study paves the way to future investigations aiming at unraveling the role of host immunity, inter-species bacterial competition and kinetics of host colonization by Bti that could be at the basis of the phenotype observed in this study.

Evidence for multiple-insecticide resistance in urban Aedes albopictus populations in southern China

BackgroundAedes albopictus (Skuse) is an invasive mosquito that has become an important vector of chikungunya, dengue and Zika viruses. In the absence of specific antiviral therapy or a vaccine, vector management is the sole method available for reducing Aedes-induced disease morbidity. Determining the resistance status of Ae. albopictus to insecticides and exploring the resistance mechanisms is essential for future vector control planning.MethodsAedes albopictus larvae and pupae were sampled from six sites (two sites each from urban, suburban and rural) in Guangzhou. The resistance bioassays were conducted against Bacillus thuringiensis israelensis (Bti): deltamethrin, propoxur and malathion for larvae; and deltamethrin, DDT, propoxur and malathion for adults. P450 monooxygenase (P450s), glutathione S-transferase (GSTs) and carboxylesterase (COEs) activities of adult mosquitoes were measured. Mutations at the knockdown resistance (kdr) gene were analyzed, and the association between kdr mutations and phenotypic resistance was tested.ResultsAdult bioassays revealed varied susceptibility against DDT, deltamethrin and propoxur in the six Ae. albopictus populations. Significantly lower mortality rates were found in urban populations than suburban and rural populations. Urban mosquito populations showed resistance against DDT, deltamethrin and propoxur, while one rural population was resistant to DDT. All populations tested were susceptible to malathion. Larval bioassays results indicated that all populations of Ae. albopictus were sensitive to the larvicide Bti and malathion. Resistance to deltamethrin and propoxur was common in larval populations. The F1534S and F1534 L mutations were found to be significantly associated with deltamethrin resistance. Biochemical assays indicated elevated detoxification enzyme activities in the field mosquito populations.ConclusionsAedes albopictus populations in Guangzhou, especially in urban areas, have developed resistance to the commonly used insecticides, primarily DDT and deltamethrin. This finding calls for resistance management and developing counter measures to mitigate the spread of resistance.

Use of Bacillus thuringiensis israelensis in integrated vector control of Aedes sp. in Sri Lanka: a prospective controlled effectiveness study.

The annual incidence of dengue has been increasing over the last few years in Sri Lanka with seasonal epidemics. Biological control of the vector has not been part of the integrated vector control implemented by the public health authorities of Sri Lanka so far. This pilot study assessed the effectiveness of using Bacillus thuringiensis israelensis (Bti) spray to control the Aedes mosquito vector population density. Prospective controlled effectiveness study in three administrative divisions of Colombo. Study areas were selected from urban, semiurban and rural administrative divisions within the district, and they were compared with two matching controls from the same division. Test areas received three cycles of Bti spraying 1 month apart. Control areas were not sprayed with Bti. Ovitrap and larval indices were calculated at baseline and 2 weeks after each spray cycle. There was a significant improvement in the adult vector population (ovitrap index) in the test areas after three spray cycles, but this effect had disappeared after 8 weeks (no residual effect). There was no consistent positive impact on larval indices (premise, container and Breteau indices) after each spray cycle. Bti might have a moderate impact on adult vector populations mediated via transient reductions in larval populations. However, this effect is not sustained probably due to rapid re-infestation. Bti spray, if implemented, can only play a supplementary role to other vector control methods.

Virulence of entomopathogenic bacteria in the bed bug, Cimex lectularius.

Due in part to the development of insecticide resistance, the common bed bug, Cimex lectularius, has overcome human intervention efforts to make a global resurgence. The failure of chemical pesticides has created a need for novel strategies to combat bed bugs. While a number of insect pests are susceptible to the use of entomopathogenic microbes or microbial-derived toxins, biological control methods have not been thoroughly explored in bed bugs. Here, we tested the virulence of three entomopathogenic bacterial species in C. lectularius to determine their potential for bed bug control. We examined bed bug survival after inoculation with live or heat-killed Serratia marcescens, Pseudomonas fluorescens, and Bacillus thuringiensis israelensis at varying temperatures. We also analyzed the viability and growth of the same bacteria in infected bed bugs. All three bacterial species were pathogenic to bed bugs. However, the effects of S. marcescens and P. fluorescens were temperature-dependent while the lethality of B. thuringiensis israelensis was not. In addition, bacterial virulence was partly dependent on the route of infection but was not strongly associated with proliferation. Thus, our results suggest multiple possible mechanisms of microbial pathogenicity in the bed bug and indicate that entomopathogenic bacteria, or products derived from them, may have useful applications for bed bug control.

Decreasing Bacillus thuringiensis israelensis sensitivity of Chironomus riparius larvae with age indicates potential environmental risk for mosquito control

Mosquito control based on the use of Bacillus thuringiensis israelensis (Bti) is regarded as an environmental friendly method. However, Bti also affects non-target chironomid midges that are recognized as a central resource in wetland food webs. To evaluate the risk for different larval stages of Chironomus riparius we performed a test series of daily acute toxicity laboratory tests following OECD guideline 235 over the entire aquatic life cycle of 28 days. Our study is the first approach that performs an OECD approved test design with Bti and C. riparius as a standard organism in ecotoxicological testing. First-instar larvae of Chironomus riparius show an increased sensitivity towards Bti which is two orders of magnitude higher than for fourth instar larvae. Most EC50 values described in the literature are based on acute toxicity tests using third and fourth instar larvae. The risk for chironomids is underestimated when applying the criteria of the biocide regulation EU 528/2012 to our data and therefore the existing assessment approval is not protective. Possible impacts of Bti induced changes in chironomid abundances and community composition may additionally affect organisms at higher trophic levels, especially in spring when chironomid midges represent a key food source for reproducing vertebrates.