scholarly journals Genome-Wide Transcriptional Analysis and Functional Validation Linked a Cluster of Epsilon Glutathione S-Transferases with Insecticide Resistance in the Major Malaria Vector Anopheles funestus across Africa

Genes ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 561
Author(s):  
Mersimine F. M. Kouamo ◽  
Sulaiman S. Ibrahim ◽  
Jack Hearn ◽  
Jacob M. Riveron ◽  
Michael Kusimo ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Expression Profiles ◽  
Resistance Management ◽  
Anopheles Funestus ◽  
Transcriptional Analysis ◽  
Malaria Vectors ◽  
Cross Resistance ◽  
Functional Validation ◽  
Genome Wide

Resistance is threatening the effectiveness of insecticide-based interventions in use for malaria control. Pinpointing genes associated with resistance is crucial for evidence-based resistance management targeting the major malaria vectors. Here, a combination of RNA-seq based genome-wide transcriptional analysis and RNA-silencing in vivo functional validation were used to identify key insecticide resistance genes associated with DDT and DDT/permethrin cross-resistance across Africa. A cluster of glutathione-S-transferase from epsilon group were found to be overexpressed in resistant populations of Anopheles funestus across Africa including GSTe1 [Cameroon (fold change, FC: 2.54), Ghana (4.20), Malawi (2.51)], GSTe2 [Cameroon (4.47), Ghana (7.52), Malawi (2.13)], GSTe3 [Cameroon (2.49), Uganda (2.60)], GSTe4 in Ghana (3.47), GSTe5 [Ghana (2.94), Malawi (2.26)], GSTe6 [Cameroun (3.0), Ghana (3.11), Malawi (3.07), Uganda (3.78)] and GSTe7 (2.39) in Ghana. Validation of GSTe genes expression profiles by qPCR confirmed that the genes are differentially expressed across Africa with a greater overexpression in DDT-resistant mosquitoes. RNAi-based knock-down analyses supported that five GSTe genes are playing a major role in resistance to pyrethroids (permethrin and deltamethrin) and DDT in An. funestus, with a significant recovery of susceptibility observed when GSTe2, 3, 4, 5 and GSTe6 were silenced. These findings established that GSTe3, 4, 5 and 6 contribute to DDT resistance and should be further characterized to identify their specific genetic variants, to help design DNA-based diagnostic assays, as previously done for the 119F-GSTe2 mutation. This study highlights the role of GSTes in the development of resistance to insecticides in malaria vectors and calls for actions to mitigate this resistance.

scholarly journals Fitness Costs of the Glutathione S-Transferase Epsilon 2 (L119F-GSTe2) Mediated Metabolic Resistance to Insecticides in the Major African Malaria Vector Anopheles Funestus

Genes ◽  
2018 ◽  
Vol 9 (12) ◽  
pp. 645 ◽  
Author(s):  
Magellan Tchouakui ◽  
Jacob M. Riveron ◽  
Doumani Djonabaye ◽  
Williams Tchapga ◽  
Helen Irving ◽  
...  
Keyword(s):  
Management Strategies ◽  
Resistance Management ◽  
Anopheles Funestus ◽  
Adult Emergence ◽  
Malaria Vectors ◽  
Adult Longevity ◽  
Heterozygote Advantage ◽  
Glutathione S Transferase ◽  
Metabolic Resistance ◽  
Life Traits

Metabolic resistance to insecticides threatens malaria control. However, little is known about its fitness cost in field populations of malaria vectors, thus limiting the design of suitable resistance management strategies. Here, we assessed the association between the glutathione S-transferase GSTe2-mediated metabolic resistance and life-traits of natural populations of Anopheles funestus. A total of 1200 indoor resting blood-fed female An. funestus (F0) were collected in Mibellon, Cameroon (2016/2017), and allowed to lay eggs individually. Genotyping of F1 mosquitoes for the L119F-GSTE2 mutation revealed that L/L119-homozygote susceptible (SS) mosquitoes significantly laid more eggs than heterozygotes L119F-RS (odds ratio (OR) = 2.06; p < 0.0001) and homozygote resistant 119F/F-RR (OR = 2.93; p < 0.0001). L/L119-SS susceptible mosquitoes also showed the higher ability for oviposition than 119F/F-RR resistant (OR = 2.68; p = 0.0002) indicating a reduced fecundity in resistant mosquitoes. Furthermore, L119F-RS larvae developed faster (nine days) than L119F-RR and L119F-SS (11 days) (X2 = 11.052; degree of freedom (df) = 4; p = 0.02) suggesting a heterozygote advantage effect for larval development. Interestingly, L/L119-SS developed faster than 119F/F-RR (OR = 5.3; p < 0.0001) revealing an increased developmental time in resistant mosquitoes. However, genotyping and sequencing revealed that L119F-RR mosquitoes exhibited a higher adult longevity compared to RS (OR > 2.2; p < 0.05) and SS (OR > 2.1; p < 0.05) with an increased frequency of GSTe2-resistant haplotypes in mosquitoes of D30 after adult emergence. Additionally, comparison of the expression of GSTe2 revealed a significantly increased expression from D1-D30 after emergence of adults (Anova test (F) = 8; df= 3; p = 0.008). The negative association between GSTe2 and some life traits of An. funestus could facilitate new resistance management strategies. However, the increased longevity of GSTe2-resistant mosquitoes suggests that an increase in resistance could exacerbate malaria transmission.

scholarly journals A differential expression of pyrethroid resistance genes in the malaria vector Anopheles funestus across Uganda is associated with patterns of gene flow

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0240743
Author(s):  
Maurice Marcel Sandeu ◽  
Charles Mulamba ◽  
Gareth D. Weedall ◽  
Charles S. Wondji
Keyword(s):  
Population Structure ◽  
Gene Flow ◽  
Genetic Structure ◽  
Genetic Differentiation ◽  
Insecticide Resistance ◽  
Pyrethroid Resistance ◽  
Anopheles Funestus ◽  
Metabolic Resistance ◽  
Transcription Analysis ◽  
Genome Wide

Background Insecticide resistance is challenging the effectiveness of insecticide-based control interventions to reduce malaria burden in Africa. Understanding the molecular basis of insecticides resistance and patterns of gene flow in major malaria vectors such as Anopheles funestus are important steps for designing effective resistance management strategies. Here, we investigated the association between patterns of genetic structure and expression profiles of genes involved in the pyrethroid resistance in An. funestus across Uganda and neighboring Kenya. Methods Blood-fed mosquitoes An. funestus were collected across the four localities in Uganda and neighboring Kenya. A Microarray-based genome-wide transcription analysis was performed to identify the set of genes associated with permethrin resistance. 17 microsatellites markers were genotyped and used to establish patterns of genetic differentiation. Results Microarray-based genome-wide transcription profiling of pyrethroid resistance in four locations across Uganda (Arua, Bulambuli, Lira, and Tororo) and Kenya (Kisumu) revealed that resistance was mainly driven by metabolic resistance. The most commonly up-regulated genes in pyrethroid resistance mosquitoes include cytochrome P450s (CYP9K1, CYP6M7, CYP4H18, CYP4H17, CYP4C36). However, expression levels of key genes vary geographically such as the P450 CYP6M7 [Fold-change (FC) = 115.8 (Arua) vs 24.05 (Tororo) and 16.9 (Kisumu)]. In addition, several genes from other families were also over-expressed including Glutathione S-transferases (GSTs), carboxylesterases, trypsin, glycogenin, and nucleotide binding protein which probably contribute to insecticide resistance across Uganda and Kenya. Genotyping of 17 microsatellite loci in the five locations provided evidence that a geographical shift in the resistance mechanisms could be associated with patterns of population structure throughout East Africa. Genetic and population structure analyses indicated significant genetic differentiation between Arua and other localities (FST>0.03) and revealed a barrier to gene flow between Arua and other areas, possibly associated with Rift Valley. Conclusion The correlation between patterns of genetic structure and variation in gene expression could be used to inform future interventions especially as new insecticides are gradually introduced.

scholarly journals CGSEA: A Flexible Tool for Evaluating the Associations of Chemicals with Complex Diseases

2020 ◽  
Vol 10 (3) ◽  
pp. 945-949
Author(s):  
Shiqiang Cheng ◽  
Mei Ma ◽  
Lu Zhang ◽  
Li Liu ◽  
Bolun Cheng ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Expression Profiles ◽  
Association Studies ◽  
Complex Diseases ◽  
Autism Spectrum ◽  
Flexible Tool ◽  
Physiological Processes ◽  
Genome Wide ◽  
Level Data

The etiology of many human complex diseases or traits involves interactions between chemicals and genes that regulate important physiological processes. It has been well documented that chemicals can contribute to disease development through affecting gene expression in vivo. In this study, we developed a flexible tool CGSEA for scanning the candidate chemicals associated with complex diseases or traits. CGSEA only need genome-wide summary level data, such as transcriptome-wide association studies (TWAS) and mRNA expression profiles. CGSEA was applied to the GWAS summaries of attention deficiency/hyperactive disorder, (ADHD), autism spectrum disorder (ASD) and cervical cancer. CGSEA identified several significant chemicals, which have been demonstrated to be involved in the development or treatment of ADHD, ASD and cervical cancer. The CGSEA program and user manual are available at https://github.com/ChengSQXJTU/CGSEA.

scholarly journals Influence of a Major Mountainous Landscape Barrier (Mount Cameroon) on the Spread of Metabolic (GSTe2) and Target-Site (Rdl) Resistance Alleles in the African Malaria Vector Anopheles funestus

Genes ◽  
2020 ◽  
Vol 11 (12) ◽  
pp. 1492
Author(s):  
Nathalie Amvongo-Adjia ◽  
Jacob M. Riveron ◽  
Flobert Njiokou ◽  
Samuel Wanji ◽  
Charles S. Wondji
Keyword(s):  
Control Strategies ◽  
Management Strategies ◽  
Resistance Management ◽  
Anopheles Funestus ◽  
Target Site ◽  
Malaria Vectors ◽  
Mount Cameroon ◽  
Parity Rate ◽  
Control Programmes ◽  
Mountainous Landscape

Increased levels of insecticide resistance in major malaria vectors such as Anopheles funestus threaten the effectiveness of insecticide-based control programmes. Understanding the landscape features impacting the spread of resistance makers is necessary to design suitable resistance management strategies. Here, we examined the influence of the highest mountain in West Africa (Mount Cameroon; 4095 m elevation) on the spread of metabolic and target-site resistance alleles in An. funestus populations. Vector composition varied across the four localities surveyed along the altitudinal cline with major vectors exhibiting high parity rate (80.5%). Plasmodium infection rates ranged from 0.79% (An. melas) to 4.67% (An. funestus). High frequencies of GSTe2R (67–81%) and RdlR (49–90%) resistance alleles were observed in An. funestus throughout the study area, with GSTe2R frequency increasing with altitude, whereas the opposite is observed for RdlR. Patterns of genetic diversity and population structure analyses revealed high levels of polymorphisms with 12 and 16 haplotypes respectively for GSTe2 and Rdl. However, the reduced diversity patterns of resistance allele carriers revealed signatures of positive selection on the two genes across the study area irrespective of the altitude. Despite slight variations associated with the altitude, the spread of resistance alleles suggest that control strategies could be implemented against malaria vectors across mountainous landscapes.

scholarly journals Identification of main malaria vectors and their insecticide resistance profile in internally displaced and indigenous communities in Eastern Democratic Republic of the Congo (DRC)

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Jeanine A. C. M. Loonen ◽  
Dominic B. Dery ◽  
Bertin Z. Musaka ◽  
Janvier B. Bandibabone ◽  
Teun Bousema ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Insecticide Resistance ◽  
Democratic Republic ◽  
Anopheles Funestus ◽  
Indigenous Communities ◽  
Health Concern ◽  
Malaria Vectors ◽  
Species Determination ◽  
Insecticide Susceptibility ◽  
Republic Of The Congo

Abstract Background Malaria remains a major public health concern in the Democratic Republic of the Congo (DRC) and its control is affected by recurrent conflicts. Médecins Sans Frontières (MSF) initiated several studies to better understand the unprecedented incidence of malaria to effectively target and implement interventions in emergency settings. The current study evaluated the main vector species involved in malaria transmission and their resistance to insecticides, with the aim to propose the most effective tools and strategies for control of local malaria vectors. Methods This study was performed in 52 households in Shamwana (Katanga, 2014), 168 households in Baraka (South Kivu, 2015) and 269 households in Kashuga (North Kivu, 2017). Anopheles vectors were collected and subjected to standardized Word Health Organization (WHO) and Center for Disease Control (CDC) insecticide susceptibility bioassays. Mosquito species determination was done using PCR and Plasmodium falciparum infection in mosquitoes was assessed by ELISA targeting circumsporozoite protein. Results Of 3517 Anopheles spp. mosquitoes collected, Anopheles gambiae sensu lato (s.l.) (29.6%) and Anopheles funestus (69.1%) were the main malaria vectors. Plasmodium falciparum infection rates for An. gambiae s.l. were 1.0, 2.1 and 13.9% for Shamwana, Baraka and Kashuga, respectively. Anopheles funestus showed positivity rates of 1.6% in Shamwana and 4.4% in Baraka. No An. funestus were collected in Kashuga. Insecticide susceptibility tests showed resistance development towards pyrethroids in all locations. Exposure to bendiocarb, malathion and pirimiphos-methyl still resulted in high mosquito mortality. Conclusions This is one of only few studies from these conflict areas in DRC to report insecticide resistance in local malaria vectors. The data suggest that current malaria prevention methods in these populations are only partially effective, and require additional tools and strategies. Importantly, the results triggered MSF to consider the selection of a new insecticide for indoor residual spraying (IRS) and a new long-lasting insecticide-treated net (LLIN). The reinforcement of correct usage of LLINs and the introduction of targeted larviciding were also included as additional vector control tools as a result of the studies.

scholarly journals Cytochrome P450 metabolic resistance (CYP6P9a) to pyrethroids imposes a fitness cost in the major African malaria vector Anopheles funestus

Heredity ◽  
2020 ◽  
Vol 124 (5) ◽  
pp. 621-632 ◽  
Author(s):  
Magellan Tchouakui ◽  
Jacob Riveron Miranda ◽  
Leon M. J. Mugenzi ◽  
Doumani Djonabaye ◽  
Murielle J. Wondji ◽  
...  
Keyword(s):  
Malaria Vector ◽  
Management Strategy ◽  
Management Strategies ◽  
Resistance Management ◽  
Anopheles Funestus ◽  
Fitness Cost ◽  
Malaria Vectors ◽  
Hybrid Strain ◽  
Metabolic Resistance ◽  
The Impact

Abstract Metabolic resistance threatens the sustainability of pyrethroid-based malaria control interventions. Elucidating the fitness cost and potential reversal of metabolic resistance is crucial to design suitable resistance management strategies. Here, we deciphered the fitness cost associated with the CYP6P9a (P450-mediated metabolic resistance) in the major African malaria vector Anopheles funestus. Reciprocal crosses were performed between a pyrethroid susceptible (FANG) and resistant (FUMOZ-R) laboratory strains and the hybrid strains showed intermediate resistance. Genotyping the CYP6P9a-R resistance allele in oviposited females revealed that CYP6P9a negatively impacts the fecundity as homozygote susceptible mosquitoes (CYP6P9a-SS) lay more eggs than heterozygote (OR = 2.04: P = 0.01) and homozygote resistant mosquitoes. CYP6P9a also imposes a significant fitness cost on the larval development as homozygote resistant larvae (CYP6P9a-RR) developed significantly slower than heterozygote and homozygote susceptible mosquitoes (χ2 = 11.2; P = 0.0008). This fitness cost was further supported by the late pupation of homozygote resistant than susceptible mosquitoes (OR = 2.50; P < 0.01). However, CYP6P9a does not impact the longevity as no difference was observed in the life span of mosquitoes with different genotypes (χ2 = 1.6; P = 0.9). In this hybrid strain, a significant decrease of the resistant CYP6P9a-RR genotype was observed after ten generations (χ2 = 6.6; P = 0.01) suggesting a reversal of P450-based resistance in the absence of selection. This study shows that the P450-mediated metabolic resistance imposes a high fitness cost in malaria vectors supporting that a resistance management strategy based on rotation could help mitigate the impact of such resistance.

scholarly journals Genome-wide comparison of phage M13-infected vs. uninfectedEscherichia coli

2005 ◽  
Vol 51 (1) ◽  
pp. 29-35 ◽  
Author(s):  
Fredrik Karlsson ◽  
Ann-Christin Malmborg-Hager ◽  
Ann-Sofie Albrekt ◽  
Carl A.K Borrebaeck
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Acid Stress ◽  
Phage Infection ◽  
Transcriptional Analysis ◽  
Filamentous Phage ◽  
Phosphotransferase System ◽  
E Coli ◽  
Genome Wide

To identify Escherichia coli genes potentially regulated by filamentous phage infection, we used oligonucleotide microarrays. Genome-wide comparison of phage M13-infected and uninfected E. coli, 2 and 20 min after infection, was performed. The analysis revealed altered transcription levels of 12 E. coli genes in response to phage infection, and the observed regulation of phage genes correlated with the known in vivo pattern of M13 mRNA species. Ten of the 12 host genes affected could be grouped into 3 different categories based on cellular function, suggesting a coordinated response. The significantly upregulated genes encode proteins involved in reactions of the energy-generating phosphotransferase system and transcription processing, which could be related to phage transcription. No genes belonging to any known E. coli stress response pathways were scored as upregulated. Furthermore, phage infection led to significant downregulation of transcripts of the bacterial genes gadA, gadB, hdeA, gadE, slp, and crl. These downregulated genes are normally part of the host stress response mechanisms that protect the bacterium during conditions of acid stress and stationary phase transition. The phage-infected cells demonstrated impaired function of the oxidative and the glutamate-dependent acid resistance systems. Thus, global transcriptional analysis and functional analysis revealed previously unknown host responses to filamentous phage infection.Key words: filamentous phage infection, global transcriptional analysis, AR, Escherichia coli.

Insecticide susceptibility status in individual species of the Anopheles gambiae complex (Diptera: Culicidae) in an area of The Gambia where pyrethroid impregnated bednets are used extensively for malaria control

1995 ◽  
Vol 85 (2) ◽  
pp. 229-234 ◽  
Author(s):  
J. Hemingway ◽  
S.W. Lindsay ◽  
G.J. Small ◽  
M. Jawara ◽  
F.H. Collins
Keyword(s):  
Insecticide Resistance ◽  
Anopheles Gambiae ◽  
Malaria Control ◽  
The Gambia ◽  
Malaria Vectors ◽  
Cross Resistance ◽  
Individual Species ◽  
Sentinel Site ◽  
Malaria Control Programme ◽  
Biochemical Assays

AbstractPyrethroid-impregnated bednets are being used nationwide in The Gambia. The future success of this malaria control programme depends partly on the vectors remaining susceptible to those insecticides used for treating the nets. The present study was carried out on the south bank of the river Gambia, during the first large scale trial of nets in this country. Thus this area represents a sentinel site for detecting insecticide resistance in local vectors. This study gives an example of how a system of early detection for resistance problems can be set up in a relatively complex situation where multiple vectors and non-vectors are present. Samples of the Anopheles gambiae complex were caught indoors using light traps in twelve villages used in the bednet study. In all villages A. gambiae sensu stricto Giles was the predominant member of the complex as determined using the rDNA-PCR diagnostic assay. Limited bioassays with DDT and permethrin, and biochemical assays for a range of insecticide resistance mechanisms suggest that the A. gambiae complex remains completely susceptible to all major classes of commonly used insecticides including pyrethroids. Biochemical assays suggest that a low frequency of DDT resistance may occur in A. melas Theobald. This is based on elevated glutathione S-transferase levels coupled with increased levels of DDT metabolism and does not involve cross-resistance to pyrethroids. Therefore we do not envisage a decline in the efficacy of treated nets against malaria vectors in the study area in the immediate future, although monitoring should be continued whilst wide-scale use of impregnated bednets is operational.

Section 4. Pyrethroid and endosulfan resistance: selection and cross resistance studies

1993 ◽  
Vol 1 ◽  
pp. 28-35 ◽  
Author(s):  
Neil W. Forrester ◽  
Matthew Cahill ◽  
Lisa J. Bird ◽  
Jacquelyn K. Layland
Keyword(s):  
Insecticide Resistance ◽  
Lower Order ◽  
Management Strategy ◽  
Pyrethroid Resistance ◽  
Resistance Management ◽  
Resistance Mechanisms ◽  
Cross Resistance ◽  
Male And Female ◽  
Insecticide Resistance Management ◽  
Resistance Selection

SummaryResistance to endosulfan and pyrethroids in Helicoverpa armigera in Australia was shown to be due to multiple rather than cross resistance. The independence of the endosulfan and pyrethroid resistance mechanisms vindicates the sequential use of these two groups in Stages I and II of the insecticide resistance management strategy, respectively. Within the cyclodienes, greatest resistance occurred to dieldrin with lower order cross resistance to endosulfan and endrin. Male and female moths expressed cyclodiene resistance equally.

scholarly journals Influence of a Major Mountainous Landscape Barrier (Mount Cameroon) on the Spread of Metabolic (GSTe2) and Target-Site (Rdl) Resistance Alleles in the African Malaria Vector Anopheles funestus

2020 ◽  
Author(s):  
Nathalie Amvongo-Adjia ◽  
Jacob M Riveron ◽  
Flobert Njiokou ◽  
Samuel Wanji ◽  
Charles S. Wondji
Keyword(s):  
Control Strategies ◽  
Management Strategies ◽  
Resistance Management ◽  
Anopheles Funestus ◽  
Target Site ◽  
Malaria Vectors ◽  
Mount Cameroon ◽  
Parity Rate ◽  
Control Programmes ◽  
Mountainous Landscape

Increased levels of insecticide resistance in major malaria vectors such as Anopheles funestus threaten the effectiveness of insecticide-based control programmes. Understanding the landscape features impacting the spread of resistance makers is necessary to design suitable resistance management strategies. Here we examined the influence of the highest mountain in West Africa (Mount Cameroon; 4,100 meters elevation) on the spread of metabolic and target-site resistance alleles in An. funestus populations. Vector composition varied across the four localities surveyed along the altitudinal cline with major vectors exhibiting high parity rate (80.5%). Plasmodium infection rates ranged from 0.79% (An. melas) to 4.67% (An. funestus). High frequencies of GSTe2R (67% - 81%) and RdlR (49% - 90%) resistance alleles were observed in An. funestus throughout the study area, with GSTe2R frequency increasing with altitude whereas the opposite is observed for RdlR. Patterns of genetic diversity and population structure analyses revealed high levels of polymorphisms with 12 and 16 haplotypes respectively for GSTe2 and Rdl. However, the reduced diversity patterns of resistance allele carriers revealed signatures of positive selection on the two genes across the study area irrespective of the altitude. Despite slight variations associated with the altitude, the spread of resistance alleles suggest that control strategies could be implemented against malaria vectors across mountainous landscapes.

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