scholarly journals Western Kenyan Anopheles gambiae showing intense permethrin resistance harbour distinct microbiota

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Diana Omoke ◽  
Mathew Kipsum ◽  
Samson Otieno ◽  
Edward Esalimba ◽  
Mili Sheth ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
High Throughput Sequencing ◽  
Pyrethroid Resistance ◽  
Sensu Stricto ◽  
Malaria Vector Control ◽  
Rrna Gene ◽  
Vector Surveillance ◽  
Bacterial Composition ◽  
Microbial Analysis

Abstract Background Insecticide resistance poses a growing challenge to malaria vector control in Kenya and around the world. Following evidence of associations between the mosquito microbiota and insecticide resistance, the microbiota of Anopheles gambiae sensu stricto (s.s.) from Tulukuyi village, Bungoma, Kenya, with differing permethrin resistance profiles were comparatively characterized. Methods Using the CDC bottle bioassay, 133 2–3 day-old, virgin, non-blood fed female F1 progeny of field-caught An. gambiae s.s. were exposed to five times (107.5 µg/ml) the discriminating dose of permethrin. Post bioassay, 50 resistant and 50 susceptible mosquitoes were subsequently screened for kdr East and West mutations, and individually processed for microbial analysis using high throughput sequencing targeting the universal bacterial and archaeal 16S rRNA gene. Results 47 % of the samples tested (n = 133) were resistant, and of the 100 selected for further processing, 99 % were positive for kdr East and 1 % for kdr West. Overall, 84 bacterial taxa were detected across all mosquito samples, with 36 of these shared between resistant and susceptible mosquitoes. A total of 20 bacterial taxa were unique to the resistant mosquitoes and 28 were unique to the susceptible mosquitoes. There were significant differences in bacterial composition between resistant and susceptible individuals (PERMANOVA, pseudo-F = 2.33, P = 0.001), with presence of Sphingobacterium, Lysinibacillus and Streptococcus (all known pyrethroid-degrading taxa), and the radiotolerant Rubrobacter, being significantly associated with resistant mosquitoes. On the other hand, the presence of Myxococcus, was significantly associated with susceptible mosquitoes. Conclusions This is the first report of distinct microbiota in An. gambiae s.s. associated with intense pyrethroid resistance. The findings highlight differentially abundant bacterial taxa between resistant and susceptible mosquitoes, and further suggest a microbe-mediated mechanism of insecticide resistance in mosquitoes. These results also indicate fixation of the kdr East mutation in this mosquito population, precluding further analysis of its associations with the mosquito microbiota, but presenting the hypothesis that any microbe-mediated mechanism of insecticide resistance would be likely of a metabolic nature. Overall, this study lays initial groundwork for understanding microbe-mediated mechanisms of insecticide resistance in African mosquito vectors of malaria, and potentially identifying novel microbial markers of insecticide resistance that could supplement existing vector surveillance tools.

scholarly journals Western Kenyan Anopheles gambiae s.s. showing intense permethrin resistance harbor distinct microbiota

2020 ◽  
Author(s):  
Diana Omoke ◽  
Mathew Kipsum ◽  
Samson Otieno ◽  
Edward Esalimba ◽  
Mili Sheth ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
High Throughput Sequencing ◽  
Pyrethroid Resistance ◽  
Malaria Vectors ◽  
Malaria Vector Control ◽  
Rrna Gene ◽  
Vector Surveillance ◽  
Anopheles Gambiae S.S ◽  
Bacterial Composition ◽  
Microbial Analysis

AbstractBackgroundInsecticide resistance poses a growing challenge to malaria vector control in Kenya and around the world. Following evidence of associations between the mosquito microbiota and insecticide resistance, we comparatively characterized the microbiota of An. gambiae s.s. from Tulukuyi village, Bungoma, Kenya, with differing permethrin resistance profiles.MethodsUsing the CDC bottle bioassay, 133 2-3 day-old, virgin, non-blood fed female F1 progeny of field-caught An. gambiae s.s. were exposed to five times (107.5μg/ml) the discriminating dose of permethrin. Post bioassay, 50 resistant and 50 susceptible mosquitoes were subsequently screened for kdr East and West mutations, and individually processed for microbial analysis using high throughput sequencing targeting the universal bacterial and archaeal 16S rRNA gene.Results47% of the samples tested (n=133) were resistant, and of the 100 selected for further processing, 99% were positive for kdr East and 1% for kdr West. Overall, 84 bacterial taxa were detected across all mosquito samples, with 36 of these shared between resistant and susceptible mosquitoes. A total of 20 were unique to the resistant mosquitoes and 28 were unique to the susceptible mosquitoes. There were significant differences in bacterial composition between resistant and susceptible individuals (F=2.33, P=0.001), with presence of Sphingobacterium, Lysinibacillus and Streptococcus (all known pyrethroid-degrading taxa), and the radiotolerant Rubrobacter, being significantly associated with resistant mosquitoes. On the other hand, the presence of Myxococcus, was significantly associated with susceptible mosquitoes.ConclusionThis is the first report of distinct microbiota in An. gambiae s.s. associated with intense pyrethroid resistance. The findings highlight differentially abundant bacterial taxa between resistant and susceptible mosquitoes, and further suggest a microbe-mediated mechanism of insecticide resistance in mosquitoes. Our results also indicate fixation of the kdr East mutation in this mosquito population, precluding further analysis of its associations with the mosquito microbiota, but presenting the hypothesis that any microbe-mediated mechanism of insecticide resistance would be likely of a metabolic nature. Overall, this study lays initial groundwork for understanding microbe-mediated mechanisms of insecticide resistance in African malaria vectors, and potentially identifying novel microbial markers of insecticide resistance that could supplement existing vector surveillance tools.

scholarly journals Investigating the relationship between insecticide resistance, underlying molecular mechanisms and malaria prevalence in Anopheles gambiae s.l. from Guinea

2018 ◽  
Author(s):  
Emma Collins ◽  
Natasha M. Vaselli ◽  
Moussa Sylla ◽  
Abdoul H. Beavogui ◽  
James Orsborne ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Molecular Mechanisms ◽  
Pyrethroid Resistance ◽  
Resistance Mechanisms ◽  
Malaria Prevalence ◽  
Sub Saharan Africa ◽  
Malaria Vector Control ◽  
Partial Restoration ◽  
Sub Saharan

AbstractThe threat of insecticide resistance across sub-Saharan Africa is anticipated to severely impact the continued effectiveness of malaria vector control. We investigated the effect of carbamate and pyrethroid resistance on Anopheles gambiae s.l age, Plasmodium falciparum infection and characterized molecular resistance mechanisms in Guinea. Pyrethroid resistance was intense, with survivors of ten times the insecticidal concentration required to kill susceptible individuals. The L1014F kdr allele was significantly associated with mosquito survival following deltamethrin or permethrin treatment (p=0.003 and p=0.04, respectively). N1575Y and I1527T mutations were identified in 13% and 10% of individuals, respectively, but neither conferred increased pyrethroid tolerance. Partial restoration of pyrethroid susceptibility following synergist pre-exposure suggest a role for mixed-function oxidases. Carbamate resistance was lower and significantly associated with the G119S Ace-1 mutation (p=0.001). Oocyst rates were 6.8% and 4.2% among resistant and susceptible mosquitoes, respectively; survivors of bendiocarb exposure were significantly more likely to be infected (p=0.03). Resistant mosquitoes had significantly lower parity rates; however, a subset of intensely pyrethroid-resistant vectors were more likely to be parous (p=0.042 and p=0.045, for survivors of five and ten times the diagnostic dose of insecticides, respectively). Our findings emphasize the need for additional studies directly assessing the influence of insecticide resistance on mosquito fitness.

scholarly journals Phenotypic, genotypic and biochemical changes during pyrethroid resistance selection in Anopheles gambiae mosquitoes

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Maxwell G. Machani ◽  
Eric Ochomo ◽  
Daibin Zhong ◽  
Guofa Zhou ◽  
Xiaoming Wang ◽  
...  
Keyword(s):  
Public Health ◽  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Selection Pressure ◽  
Pyrethroid Resistance ◽  
Natural Populations ◽  
Biochemical Changes ◽  
Moderate Intensity ◽  
Agricultural System ◽  
Selection For

Abstract The directional selection for insecticide resistance due to indiscriminate use of insecticides in public health and agricultural system favors an increase in the frequency of insecticide-resistant alleles in the natural populations. Similarly, removal of selection pressure generally leads to decay in resistance. Past investigations on the emergence of insecticide resistance in mosquitoes mostly relied on field survey of resistance in vector populations that typically had a complex history of exposure to various public health and agricultural pest control insecticides in nature, and thus the effect of specific insecticides on rate of resistance emergency or resistance decay rate is not known. This study examined the phenotypic, genotypic, and biochemical changes that had occurred during the process of selection for pyrethroid resistance in Anopheles gambiae, the most important malaria vector in Africa. In parallel, we also examined these changes in resistant populations when there is no selection pressure applied. Through repeated deltamethrin selection in adult mosquitoes from a field population collected in western Kenya for 12 generations, we obtained three independent and highly pyrethroid-resistant An. gambiae populations. Three susceptible populations from the same parental population were generated by removing selection pressure. These two lines of mosquito populations differed significantly in monooxygenase and beta-esterase activities, but not in Vgsc gene mutation frequency, suggesting metabolic detoxification mechanism plays a major role in generating moderate-intensity resistance or high-intensity resistance. Pre-exposure to the synergist piperonyl butoxide restored the susceptibility to insecticide among the highly resistant mosquitoes, confirming the role of monooxygenases in pyrethroid resistance. The rate of resistance decay to become fully susceptible from moderate-intensity resistance took 15 generations, supporting at least 2-years interval is needed when the rotational use of insecticides with different modes of action is considered for resistance management.

scholarly journals Fine scale spatial investigation of multiple insecticide resistance and underlying target-site and metabolic mechanisms in Anopheles gambiae in central Côte d’Ivoire

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Welbeck A. Oumbouke ◽  
Patricia Pignatelli ◽  
Antoine M. G. Barreaux ◽  
Innocent Z. Tia ◽  
Alphonsine A. Koffi ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Cote D'ivoire ◽  
Côte D’Ivoire ◽  
Pyrethroid Resistance ◽  
Controlled Trial ◽  
Management Strategies ◽  
Cluster Randomised Controlled Trial ◽  
Cote D’Ivoire ◽  
Côte D'ivoire

Abstract Routine monitoring of occurrence, levels and mechanisms of insecticide resistance informs effective management strategies, and should be used to assess the effect of new tools on resistance. As part of a cluster randomised controlled trial evaluating a novel insecticide-based intervention in central Côte d’Ivoire, we assessed resistance and its underlying mechanisms in Anopheles gambiae populations from a subset of trial villages. Resistance to multiple insecticides in An. gambiae s.s. and An. coluzzii was detected across villages, with dose–response assays demonstrating extremely high resistance intensity to the pyrethroid deltamethrin (> 1,500-fold), and mortality following exposure to pyrethroid-treated bednets was low (< 30% mortality in cone bioassays). The 1014F kdr mutation was almost fixed (≥ 90%) in all villages but the 1575Y kdr-amplifying mutation was relatively rare (< 15%). The carbamate and organophosphate resistance-associated Ace-1 G119S mutation was also detected at moderate frequencies (22–43%). Transcriptome analysis identified overexpression of P450 genes known to confer pyrethroid resistance (Cyp9K1, Cyp6P3, and Cyp6M2), and also a carboxylesterase (COEAE1F) as major candidates. Cyp6P3 expression was high but variable (up to 33-fold) and correlated positively with deltamethrin resistance intensity across villages (r2 = 0.78, P = 0.02). Tools and strategies to mitigate the extreme and multiple resistance provided by these mechanisms are required in this area to avoid future control failures.

scholarly journals PCR-RFLP Detection and Genogroup Identification of Piscirickettsia salmonis in Field Samples

Pathogens ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 358
Author(s):  
Pamela Aravena ◽  
Rodrigo Pulgar ◽  
Javiera Ortiz-Severín ◽  
Felipe Maza ◽  
Alexis Gaete ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
In Silico Analysis ◽  
Cost Effective ◽  
Rrna Gene ◽  
Bacterial Composition ◽  
Bacterial Dna ◽  
Phenotypic Differences ◽  
Field Samples ◽  
Pcr Rflp ◽  
Digestion Pattern

Piscirickettsia salmons, the causative agent of piscirickettsiosis, is genetically divided into two genomic groups, named after the reference strains as LF-89-like or EM-90-like. Phenotypic differences have been detected between the P. salmonis genogroups, including antibiotic susceptibilities, host specificities and pathogenicity. In this study, we aimed to develop a rapid, sensitive and cost-effective assay for the differentiation of the P. salmonis genogroups. Using an in silico analysis of the P. salmonis 16S rDNA digestion patterns, we have designed a genogroup-specific assay based on PCR-restriction fragment length polymorphism (RFLP). An experimental validation was carried out by comparing the restriction patterns of 13 P. salmonis strains and 57 field samples obtained from the tissues of dead or moribund fish. When the bacterial composition of a set of field samples, for which we detected mixtures of bacterial DNA, was analyzed by a high-throughput sequencing of the 16S rRNA gene amplicons, a diversity of taxa could be identified, including pathogenic and commensal bacteria. Despite the presence of mixtures of bacterial DNA, the characteristic digestion pattern of the P. salmonis genogroups could be detected in the field samples without the need of a microbiological culture and bacterial isolation.

scholarly journals Pyrethroid Resistance in Anopheles gambiae, in Bomi County, Liberia, Compromises Malaria Vector Control

PLoS ONE ◽  
2012 ◽  
Vol 7 (9) ◽  
pp. e44986 ◽  
Author(s):  
Emmanuel A. Temu ◽  
Caroline Maxwell ◽  
Godwil Munyekenye ◽  
Annabel F. V. Howard ◽  
Stephen Munga ◽  
...  
Keyword(s):  
Vector Control ◽  
Anopheles Gambiae ◽  
Malaria Vector ◽  
Pyrethroid Resistance ◽  
Malaria Vector Control

scholarly journals Anopheles gambiae and Anopheles arabiensis population densities and infectivity in Kopere village, Western Kenya

2012 ◽  
Vol 6 (08) ◽  
pp. 637-643 ◽  
Author(s):  
Andrew Ambogo Obala ◽  
Helen L Kutima ◽  
Henry D.N. Nyamogoba ◽  
Anne W Mwangi ◽  
Chrispinus J Simiyu ◽  
...  
Keyword(s):  
Anopheles Gambiae ◽  
Malaria Vector ◽  
Anopheles Arabiensis ◽  
Mosquito Species ◽  
Sensu Stricto ◽  
Malaria Vector Control ◽  
Insecticide Treated Nets ◽  
Western Kenya ◽  
Endemic Malaria ◽  
Vector Mosquito

Introduction: This study was conducted in a sugar belt region of western Kenya interfacing epidemic and endemic malaria transmission. We investigated Anopheles gambiae sensu stricto (ss) and Anopheles arabiensis species compositions and densities, human host choice, and infectivity. Methodology: Mosquitoes were captured using pyrethrum spray catch technique and first identified based on morphology; species were confirmed by PCR. Blood meal preference and sporozoite rates were determined by ELISA. Parity rates and entomological inoculation rates (EIR) were determined. Seasonal densities were compared against environmental temperatures, relative humidity and rainfall. Results: In total 2,426 An. gambiae were collected.  Out of 1,687 female blood-fed mosquitoes, 272 were randomly selected for entomological tests. An. gambiae ss and An. arabiensis comprised 75% (205/272) and 25% (68/272) of the selection, respectively. An. gambiae ss had higher preference for human blood (97%; n=263/272) compared with An. arabiensis, which mostly fed on bovines (88%; n=239/272).  The sporozoite and parity rates were 6% (16/272) and 66% (179/272) for An. gambiae ss and 2% (4/272) and 53% (144/272) for An. arabiensis respectively, while EIR was 0.78 infective bites/person/night.  Climate (ANOVA; F=14.2; DF=23) and temperature alone (r=0.626; t=3.75; p=0.001) were significantly correlated with vector densities. Conclusion: An. gambiae ss are the most efficient malaria vector mosquito species in Kopere village. Because An. gambiae ss largely rests and feeds indoors, use of indoor residual spray and insecticide-treated nets is likely the most suitable approach to malaria vector control in Kopere village and other parts of Kenya where this species is abundant. 

scholarly journals Functional genetic validation of key genes conferring insecticide resistance in the major African malaria vector, Anopheles gambiae

2019 ◽  
Author(s):  
Adriana Adolfi ◽  
Beth Poulton ◽  
Amalia Anthousi ◽  
Stephanie Macilwee ◽  
Hilary Ranson ◽  
...  
Keyword(s):  
Public Health ◽  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Pyrethroid Resistance ◽  
Control Strategies ◽  
Resistance Mechanisms ◽  
Effective Control ◽  
Cross Resistance ◽  
Toxic Metabolite ◽  
Control Programs

ABSTRACTResistance in Anopheles gambiae to members of all four major classes (pyrethroids, carbamates, organochlorines and organophosphates) of public health insecticides limits effective control of malaria transmission in Africa. Increased expression of detoxifying enzymes has been associated with resistance, but direct functional validation in An. gambiae has been lacking. Here we perform transgenic analysis using the GAL4/UAS system to examine insecticide resistance phenotypes conferred by increased expression of the three genes - Cyp6m2, Cyp6p3 and Gste2 - most often found upregulated in resistant An. gambiae. We report the first evidence in An. gambiae that organophosphate and organochlorine resistance is conferred by overexpression of GSTE2 in a broad tissue profile. Pyrethroid and carbamate resistance is bestowed by similar Cyp6p3 overexpression, and Cyp6m2 confers only pyrethroid resistance when overexpressed in the same tissues. Conversely, such Cyp6m2 overexpression increases susceptibility to the organophosphate malathion, presumably due to conversion to a more toxic metabolite. No resistant phenotypes are conferred when either Cyp6 gene overexpression is restricted to the midgut or oenocytes, answering long standing questions related to the importance of these tissues in resistance to contact insecticides. Validation of genes conferring resistance provides markers to guide control strategies, and the observed negative cross-resistance due to Cyp6m2 gives credence to proposed dual insecticide strategies to overcome pyrethroid resistance. These trasnsgenic An. gambiae resistant lines are being used to test potential liabilities in new active compounds early in development.SIGNIFICANCE STATEMENTInsecticide resistance in Anopheles gambiae mosquitoes can derail malaria control programs, and to overcome it we need to discover the underlying molecular basis. Here, for the first time, we characterise three genes most often associated with insecticide resistance directly by their overproduction in genetically modified An. gambiae. We show that overexpression of each gene confers resistance to representatives of at least one insecticide class and, taken together, the three genes provide cross-resistance to all four major insecticide classes currently used in public health. These data validate the candidate genes as markers to monitor the spread of resistance in mosquito populations. The modified mosquitoes produced are also valuable tools to pre-screen new insecticides for potential liabilities to existing resistance mechanisms.

scholarly journals Durability of three types of dual active ingredient long-lasting insecticidal net compared to a pyrethroid-only LLIN in Tanzania: protocol for a prospective cohort study nested in a cluster randomized controlled trial

2021 ◽  
Author(s):  
Jackline L Martin ◽  
Louisa A Messenger ◽  
Franklin W Mosha ◽  
Eliud Lukole ◽  
Jacklin F Mosha ◽  
...  
Keyword(s):  
Active Ingredient ◽  
Pyrethroid Resistance ◽  
Controlled Trial ◽  
Cluster Randomised Controlled Trial ◽  
Blood Feeding ◽  
Sensu Stricto ◽  
Malaria Vector Control ◽  
Pyrethroid Insecticides ◽  
Egg Maturation ◽  
Randomised Controlled

Introduction: Progress achieved by long-lasting insecticidal nets (LLINs) against malaria is threatened by the widespread selection of pyrethroid resistance among vector populations. LLINs with non-pyrethroid insecticides are urgently needed. This study aims to assess the durability of three novel dual active ingredient LLINs and to parameterise a mathematical model to predict epidemiological outcomes of these products for malaria vector control. Methods: A WHO Phase 3 active ingredients and textile durability study will be carried out within a cluster randomised controlled trial in Misungwi district in 40 clusters. The following treatments will be evaluated; 1/ Interceptor G2 combining chlorfenapyr and a pyrethroid alpha-cypermethrin, 2/ Royal Guard treated with pyriproxyfen and alpha-cypermethrin and 3/ Olyset Plus which incorporates a synergist piperonyl butoxide and the pyrethroid permethrin, and 4/ a reference standard pyrethroid-only LLIN (Interceptor). 750 nets will be followed in 5 clusters per intervention arm at 6, 12, 24 and 36 months post distribution for survivorship and hole index assessment. A second cohort of 1950 nets per each type will be identified in 10 clusters, of which 30 LLINs will be withdrawn for bio-efficacy and chemical analysis every 6 months up to 36 months and another 30 collected for an experimental hut trial study every year. Bio-efficacy will be assessed using cone bioassays and tunnel tests against susceptible and resistant laboratory strains of Anopheles gambiae sensu stricto. Efficacy of field collected nets will be compared in six experimental huts. The main outcomes will be Anopheles mortality up to 72 hours post exposure, blood feeding and egg maturation using ovary dissection to assess impact on fecundity. Ethics and dissemination: Ethical approval was received from the Tanzanian ethics review committee as well as from each institution. Study findings will be disseminated via reports and presentations to national and international stakeholders, conferences, and peer-reviewed publications.

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