Field efficacy of pyrethroid treated plastic sheeting (durable lining) in combination with long lasting insecticidal nets against malaria vectors

BACKGROUND: Insecticide treated plastic sheeting (ITPS), sometimes known as durable lining, has potential as a long-lasting insecticidal surface for malaria vector control when used as lining for interior walls and ceilings inside the home. Against a backdrop of increasing long lasting net (LN) coverage, we examined the effect of combining permethrin-treated plastic sheeting (ITPS) with LNs in Burkina Faso. METHODS: A verandah trap experimental hut trial of ITPS with or without Olyset LN was conducted in the Vallée du Kou near Bobo-Dioulasso, where the two molecular forms of Anopheles gambiae s.s., S (frequency 65%) and M (frequency 35%), occur. The S form is mostly pyrethroid resistant (Fkdr = 92%) owing to the kdr mechanism, and the M form is mostly kdr susceptible (Fkdr = 7%). The treatment arms included ITPS, Olyset, ITPS plus Olyset, ITPS plus untreated net (with or without holes), and untreated control. RESULTS: ITPS was significantly inferior to Olyset LN in terms of mortality (37% vs 63%), blood feeding inhibition (20% vs 81%) and deterrence (0 vs 42%) effects, and hence altogether inferior as a means of personal protection (16% vs 89%). The addition of ITPS to Olyset did not improve mortality (62%), blood feeding inhibition (75%), deterrence (50%) or personal protection (88%) over that of Olyset used alone. Use of untreated nets - both holed and intact - with ITPS provided greater protection from blood-feeding. The intact net/ITPS combination killed more mosquitoes than ITPS on its own. CONCLUSIONS: Although ITPS has a potential role for community control of malaria, at low coverage it is unlikely to be as good as Olyset LNs for household protection. The combination of pyrethroid IRS and pyrethroid LN - as practiced in some countries - is unlikely to be additive except, perhaps, at high levels of IRS coverage. A combination of LN and ITPS treated with an alternative insecticide is likely to be more effective, particularly in areas of pyrethroid resistance

Inversion genotyping in the Anopheles gambiae complex using high-throughput array and sequencing platforms

Chromosomal inversion polymorphisms have special importance in the Anopheles gambiae complex of malaria vector mosquitoes, due to their role in local adaptation and range expansion. The study of inversions in natural populations is reliant on polytene chromosome analysis by expert cytogeneticists, a process that is limited by the rarity of trained specialists, low throughput, and restrictive sampling requirements. To overcome this barrier, we ascertained tag single nucleotide polymorphisms (SNPs) that are highly correlated with inversion status (inverted or standard orientation). We compared the performance of the tag SNPs using two alternative high throughput molecular genotyping approaches vs. traditional cytogenetic karyotyping of the same 960 individual An. gambiae and An. coluzzii mosquitoes sampled from Burkina Faso, West Africa. We show that both molecular approaches yield comparable results, and that either one performs as well or better than cytogenetics in terms of genotyping accuracy. Given the ability of molecular genotyping approaches to be conducted at scale and at relatively low cost without restriction on mosquito sex or developmental stage, molecular genotyping via tag SNPs has the potential to revitalize research into the role of chromosomal inversions in the behavior and ongoing adaptation of An. gambiae and An. coluzzii to environmental heterogeneities

A PCR-RFLP method for genotyping of inversion 2Rc in Anopheles coluzzii

Background: Genotyping of polymorphic chromosomal inversions in malaria vectors such as An. coluzzii Coetzee & Wilkerson is important, both because they cause cryptic population structure that can mislead vector analysis and control and because they influence epidemiologically relevant eco-phenotypes. The conventional cytogenetic method of genotyping is an impediment because it is labor intensive, requires specialized training, and can be applied only to one gender and developmental stage. Here, we circumvent these limitations by developing a simple and rapid molecular method of genotyping inversion 2Rc in An. coluzzii that is both economical and field-friendly. This inversion is strongly implicated in temporal and spatial adaptations to climatic and ecological variation, particularly aridity. Methods: Using a set of tag single-nucleotide polymorphisms (SNPs) strongly correlated with inversion orientation, we identified those that overlapped restriction enzyme recognition sites and developed four polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP) assays that distinguish alternative allelic states at the tag SNPs. We assessed the performance of these assays using mosquito population samples from Burkina Faso that had been cytogenetically karyotyped as well as genotyped, using two complementary high-throughput molecular methods based on tag SNPs. Further validation was performed using mosquito population samples from additional West African (Benin, Mali, Senegal) and Central African (Cameroon) countries. Results: Of four assays tested, two were concordant with the 2Rc cytogenetic karyotype > 90% of the time in all samples. We recommend that these two assays be employed in tandem for reliable genotyping. By accepting only those genotypic assignments where both assays agree, > 99% of assignments are expected to be accurate. Conclusions: We have developed tandem PCR-RFLP assays for the accurate genotyping of inversion 2Rc in An. coluzzii. Because this approach is simple, inexpensive, and requires only basic molecular biology equipment, it is widely accessible. These provide a crucial tool for probing the molecular basis of eco-phenotypes relevant to malaria epidemiology and vector control. [Figure not available: see fulltext.

Development of reference transcriptomes for the major field insect pests of cowpea: a toolbox for insect pest management approaches in West Africa

Cowpea is a widely cultivated and major nutritional source of protein for many people that live in West Africa. Annual yields and longevity of grain storage is greatly reduced by feeding damage caused by a complex of insect pests that include the pod sucking bugs, Anoplocnemis curvipes Fabricius (Hemiptera: Coreidae) and Clavigralla tomentosicollis Stål (Hemiptera: Coreidae); as well as phloem-feeding cowpea aphids, Aphis craccivora Koch (Hemiptera: Aphididae) and flower thrips, Megalurothrips sjostedti Trybom (Thysanoptera: Thripidae). Efforts to control these pests remain a challenge and there is a need to understand the structure and movement of these pest populations in order to facilitate the development of integrated pest management strategies (IPM). Molecular tools have the potential to help facilitate a better understanding of pest populations. Towards this goal, we used 454 pyrosequencing technology to generate 319,126, 176,262, 320,722 and 227,882 raw reads from A. curvipes, A. craccivora, C. tomentosicollis and M. sjostedti, respectively. The reads were de novo assembled into 11,687, 7,647,10,652 and 7,348 transcripts for A. curvipes, A. craccivora, C. tomentosicollis and M. sjostedti, respectively.Functional annotation of the resulting transcripts identified genes putatively involved in insecticide resistance, pathogen defense and immunity. Additionally, sequences that matched the primary aphid endosymbiont, Buchneraaphidicola, were identified among A. craccivora transcripts. Furthermore, 742, 97, 607 and 180 single nucleotide polymorphisms (SNPs) were respectively predicted among A. curvipes, A. craccivora, C. tomentosicollis and M.sjostedti transcripts, and will likely be valuable tools for future molecular genetic marker development. These results demonstrate that Roche 454-based transcriptome sequencing could be useful for the development of genomic resources for cowpea pest insects in West Africa

Target-site resistance mutations (kdr and RDL), but not metabolic resistance, negatively impact male mating competiveness in the malaria vector Anopheles gambiae

The implementation of successful insecticide resistance management strategies for malaria control is currently hampered by poor understanding of the fitness cost of resistance on mosquito populations, including their mating competiveness. To fill this knowledge gap, coupled and uncoupled Anopheles gambiae s.l. males (all M form (Anopheles coluzzii)) were collected from mating swarms in Burkina Faso. This multiple insecticide resistant population exhibited high 1014F kdrR allele frequencies (460%) and RDLR (480%) in contrast to the Ace-1R allele (o6%). Kdr heterozygote males were more likely to mate than homozygote resistant (OR=2.36; Po0.001), suggesting a negative impact of kdr on An. coluzzii mating ability. Interestingly, heterozygote males were also more competitive than homozygote susceptible (OR=3.26; P=0.006), suggesting a heterozygote advantage effect. Similarly, heterozygote RDLR/RDLS were also more likely to mate than homozygote-resistant males (OR=2.58; P=0.007). Furthermore, an additive mating disadvantage was detected in male homozygotes for both kdr/RDL-resistant alleles. In contrast, no fitness difference was observed for the Ace-1 mutation. Comparative microarray-based genome-wide transcription analysis revealed that metabolic resistance did not significantly alter the mating competitiveness of male An. coluzzii mosquitoes. Indeed, no significant difference of expression levels was observed for the main metabolic resistance genes, suggesting that metabolic resistance has a limited impact on male mating competiveness. In addition, specific gene classes/GO terms associated with mating process were detected including sensory perception and peroxidase activity. The detrimental impact of insecticide resistance on mating competiveness observed here suggests that resistance management strategies such as insecticide rotation could help reverse the resistance, if implemented early

Diapause disruption in Cirina butyrospermi Vuillet (Lepidoptera, Attacidae), the shea caterpillar, in Burkina Faso

The shea caterpillar Cirina butyrospermi is an important insect, highly valued as a human food item in Burkina Faso. However, its appearance is seasonal due to its univoltine cycle. This study therefore investigated the possibilities of breaking the nymphal diapause by changing the environmental factors and through the hormonal treatment of prepupae and pupae using bovine insulin and 20-hydroxyecdysone. Changes in humidity and temperature did not result in emergence, suggesting a mandatory nature of the diapause in C. butyrospermi. Injection of 20-hydroxyecdysone between 20 and 40 ng on 20 C. butyrospermi pupae resulted in 15.24 and 47.5% emergence, respectively. The incubation time varied between 40 and 38 days, respectively. No emergence was observed with the injection of bovine insulin. Dipping of C. butyrospermi larvae and pupae in solutions of 20-hydroxyecdysone resulted in similar rates of emergence between the two stages, with slight variations between individual doses: (1) for larvae, emergence was recorded at 10, 8, 5 and 15 mg/l with 98.5, 62.14, 25.73 and 24.16%, respectively; the incubation times varied from 39 days at 5 mg/l to 26 days at 20 mg/l; and (2) for pupae, emergence occurred between 5 and 20 mg/l, with the highest emergence rate recorded at 10, 8 and 15 mg/l with 94.58, 65.83 and 29.58%, respectively; the incubation times varied from 53 days for the lowest dose (5 mg/l) to 37 days (20 mg/l); the best emergence rate of 94.58% coincided with an incubation time of 43 days at 10 mg/l. No emergence was observed beyond 20 mg/l in both stages. Hormonal treatment with 20-hydroxyecdysone did not affect the fecundity of C. butyrospermi, with the fecundity of artificially emerging adults overlapping with that of naturally emerging adults. The emergence rate for both was similar. These results contribute to a better understanding of the physiology of this insect, constituting a breakthrough in its sustainable use as human food

Feeding preferences of the legume pod borer Maruca vitrata (Lepidoptera: Crambidae) larvae and suitability of different flower parts for larval development

With the advent of transgenic Bacillus thuringiensis (Bt)-cowpea, there is a need to identify the feeding preferences of Maruca vitrata Fab. to determine in which component of the plant the expression of the toxin needs to be the highest to ensure the greatest efficacy of insecticidal Bt proteins. In the current study, we examined the feeding preferences of M. vitrata larvae in a naturally infested cowpea field. We also tested, in the laboratory, the suitability of different flower components for the larval development of M. vitrata. Our results indicate that in the field, all types of flowers, regardless of their age, were infested with M. vitrata larvae. The reproductive organs, in the flowers, were the preferred feeding diet for the larvae. Laboratory bioassays confirmed that the reproductive organs were the optimal tissues for M. vitrata larval development. The implications of these findings for transgenic Bt-cowpea are discussed

Modeling Evolution of Resistance by Maruca vitrata (Lepidoptera: Crambidae) to Transgenic Insecticidal Cowpea in Africa

We created a detailed model of the Maruca vitrata (F.) and cowpea [Vigna unguiculata (L.) Walp] system to study the possible evolution of resistance by the insect to transgenic insecticidal cowpea, which is under development. We focused on population dynamics and genetics in a region of west Africa. We simulated single-toxin and pyramided (two-toxin) cowpea and emphasized conservative, worst-case scenarios in our analysis. The results indicate that as long as a pyramided, transgenic cowpea can be developed, seed saving by farmers and reliance on natural refuge are not major problems for resistance management. Furthermore, it is possible that one or both toxins in the pyramid may not need to be high dose for evolution to be delayed significantly (>20 yr or 80 generations for resistance to become a concern if transgenic cowpea is deployed in areas where M. vitrata is endemic). If efforts are made to deploy transgenic cowpea only into the regions where M. vitrata is not endemic, then there is little to no concern with resistance emerging in the M. vitrata populatio

How do Service Providers and Clients Perceive Interorganizational Networks?

Interorganizational networks are important structures for both service providers, who must navigate them as part of their organizational roles, and clients, who use them for the purposes of receiving benefits. This research develops and tests a conceptual model that explains some of the differences in the ways that these two groups perceive these networks. Drawing on surveys/interviews with 200 clients and 63 service providers of agricultural development training in Burkina Faso, this research demonstrates that clients perceive interorganizational networks differently than service providers. In particular, these results demonstrate that service providers perceive more organizations in the network, more competitive and collaborative ties among those organizations, and more competitive ties per organization than clients. From these results, we draw implications for social network, development communication, and organizational fields’ research

Costs of insensitive acetylcholinesterase insecticide resistance for the malaria vector Anopheles gambiae homozygous for the G119S mutation

<p>Abstract</p> <p>Background</p> <p>The G119S mutation responsible for insensitive acetylcholinesterase resistance to organophosphate and carbamate insecticides has recently been reported from natural populations of <it>Anopheles gambiae </it>in West Africa. These reports suggest there are costs of resistance associated with this mutation for <it>An. gambiae</it>, especially for homozygous individuals, and these costs could be influential in determining the frequency of carbamate resistance in these populations.</p> <p>Methods</p> <p>Life-history traits of the AcerKis and Kisumu strains of <it>An. gambiae </it>were compared following the manipulation of larval food availability in three separate experiments conducted in an insecticide-free laboratory environment. These two strains share the same genetic background, but differ in being homozygous for the presence or absence of the G119S mutation at the <it>ace-1 </it>locus, respectively.</p> <p>Results</p> <p>Pupae of the resistant strain were significantly more likely to die during pupation than those of the susceptible strain. Ages at pupation were significantly earlier for the resistant strain and their dry starved weights were significantly lighter; this difference in weight remained when the two strains were matched for ages at pupation.</p> <p>Conclusions</p> <p>The main cost of resistance found for <it>An. gambiae </it>mosquitoes homozygous for the G119S mutation was that they were significantly more likely to die during pupation than their susceptible counterparts, and they did so across a range of larval food conditions. Comparing the frequency of G119S in fourth instar larvae and adults emerging from the same populations would provide a way to test whether this cost of resistance is being expressed in natural populations of <it>An. gambiae </it>and influencing the dynamics of this resistance mutation.</p