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Yalwala, S.

Publications and source records attributed to Yalwala, S..

6 recordsLinked to original sources

Pyrethroid Resistance Status and Multiple kdr Mutations (F1534C/L) in Aedes aegypti Populations from Zika-prone Areas in Lamu County, Kenya

Aedes aegypti mosquitoes are the primary vectors for dengue, yellow fever, chikungunya and zika virus transmission, posing significant public health risks. In Kenya, these viruses drive disease outbreaks especially, dengue and chikungunya with coastal Kenya being the most affected. Low-level circulation of the Zika virus has been reported in parts of the Kenyan coast, with confirmed cases reported in Lamu County between August to September 2024. Except for yellow fever, there are no approved vaccines or therapeutics, hence vector control remains the most effective means of protection. However, prolonged exposure to insecticides can lead to resistance, threatening these interventions. Therefore, monitoring resistance in mosquito populations is critical to allow for appropriate interventions using effective chemical classes to prevent disease outbreaks. This study aimed to establish the levels of resistance to pyrethroids, and associated markers, among Ae. aegypti populations in sections of Lamu County where there had been a recent localized outbreak of Zika. Mosquito eggs were collected from Mkomani, Kashmir, and Kandahar villages in Lamu County, reared and tested for susceptibility to three pyrethroid insecticides (0.75% permethrin, 0.05% Alpha-cypermethrin and 0.05% deltamethrin) using WHO tube assays. Genotyping of knockdown resistance (kdr) mutations L982W, S989P, A1007G, V1016G/I, and F1534C was done using Sanger sequencing. Association between resistant phenotypes and genotypes were inferred. The results varied between the three pyrethroids with high resistance to permethrin observed (6-15% mortality), for deltamethrin mortality ranged between 53-57%, while for alphacypermethrin 88%-99% mortality was observed. Two mutation types and six genotypes were identified at F1534. No other kdr mutations were detected. The CC genotype was significantly associated with 0.75% permethrin resistance in Ae. aegypti populations (OR = 2.87, 95% CI: 1.34-6.17, P = 0.0036). The current data show that Ae. aegypti from Kandahar, Kashmir and Mkomani villages in Lamu County have developed very high resistance to permethrin, and varying resistance to other pyrethroids, thus threatening pyrethroid-based control strategies in this region, highlighting the need for alternative strategies to control the vector for arboviruses.

evolutionary biology↗

Metagenomic Characterization of Aedes aegypti Virome in Kwale County, Kenya

BackgroundAedes aegypti is a primary vector of arboviruses, including dengue, chikungunya, yellow fever, and Zika, and is widespread along the Kenyan coast, a region with recurrent outbreaks. In addition to human-pathogenic arboviruses, Ae. aegypti harbors insect-specific viruses (ISVs) that replicate exclusively in arthropods and may influence mosquito physiology, immunity, and vector competence. However, data on ISVs in Kenyan Ae. aegypti populations are limited. MethodsTwenty-nine mosquito pools containing 20 individual mosquitoes from Kwale County were homogenized, combined into a superpool, and subjected to total RNA extraction. Libraries were sequenced on an Illumina MiSeq platform. Initial analysis was done on CZ-ID platform. Reads were quality-filtered using PrinseqLite and assembled de novo with MEGAHIT. Phylogenetic analyses was performed with IQ-TREE using the Maximum Likelihood method. ResultsMetagenomic analysis revealed diverse ISVs in Ae. aegypti. Complete genomes of Fako virus and Tesano Aedes virus, and partial genomes of Aedes partiti-like virus, Cell fusing agent virus, and Formosus virus, were recovered. Genome lengths ranged from 1,149 to 10,146 nucleotides, with 91-99.5% identity to reference strains. Phylogenetic analysis placed the viruses within established ISV lineages, showing close evolutionary relationships with strains previously reported from Africa and other regions. ConclusionsThis study provides comprehensive characterization of ISVs in Ae. aegypti from Kwale County. The recovery of complete and near-complete genomes demonstrates the diversity and active circulation of ISVs, establishing a baseline for future studies on mosquito viromes, virus-mosquito interactions, and potential impacts on vector competence. ImportanceInsect-specific viruses (ISVs) are widespread in mosquitoes and replicate exclusively in arthropods, influencing mosquito physiology, immunity, and potentially vector competence for human-pathogenic arboviruses. Despite recurrent arboviral outbreaks along the Kenyan coast, the diversity and ecological roles of ISVs in Aedes aegypti remain poorly understood. This study provides a comprehensive metagenomic characterization of ISVs in Ae. aegypti from Kwale County, revealing complete genomes of Fako virus and Tesano Aedes virus, along with near-complete genomes of Aedes partiti-like virus, Cell fusing agent virus, and Formosus virus. These findings establish a baseline for mosquito virome composition in this region and underscore the active circulation of diverse ISVs in natural populations. Understanding these virus-mosquito interactions is critical for interpreting arbovirus ecology, assessing the potential impacts of ISVs on vector competence, and informing future vector surveillance and biological control strategies.

genomics↗

Detection and Characterization of West Nile Virus with evidence of Transovarial Transmission in the Coastal region, Kenya

BackgroundWest Nile virus (WNV) is a mosquito-borne flavivirus of global public health significance, maintained in an enzootic cycle between birds and mosquitoes, with humans and other mammals as incidental hosts. Understanding WNV circulation in diverse mosquito populations is critical for predicting and mitigating outbreaks. This study investigated mosquito populations from coastal Kenya, where WNV was detected, genetically characterized, and evidence of transovarial transmission in Aedes aegypti was observed. MethodsMosquitoes were collected from Kwale, Kilifi, Mombasa, and Isiolo counties (n=14,105) and pooled by species and location (1,596 pools). Pools were inoculated on Vero E6 cells, followed by RNA extraction, Illumina MiSeq sequencing, and preliminary analysis on CZ-ID. Reads were quality-controlled (PrinseqLite v0.20.4), assembled de novo (MEGAHIT v1.2.9), and analyzed via BLAST. Phylogenetic reconstruction used Maximum Likelihood, and codon-level selection pressure was evaluated using FEL, MEME, and FUBAR on Datamonkey. ResultsWNV was detected in ten pools: eight Lineage 1a and two Lineage 2. Virus isolates came from Culex pipiens, Culex univittatus, Anopheles funestus, Aedes aegypti, and Eretmapodites chrysogaster. Notably, one Lineage 1a isolate from a male Aedes aegypti confirmed transovarial transmission. Six codons were under diversifying selection, NS2B gene was found to carry the V103A mutation. ConclusionWNV in coastal Kenyan mosquitoes revealed transovarial transmission in Aedes aegypti. Lineage-specific evolution, codons under positive selection, and the NS2B:V103A mutation demonstrate ongoing viral adaptation. These findings highlight the need for continued genomic surveillance and targeted vector studies to guide WNV control strategies in Kenya and the wider region. ImportanceWest Nile virus (WNV) remains a globally important arbovirus, yet genomic and experimental data from under-sampled regions such as coastal East Africa are limited. This study provides the first integrated molecular and genotypic characterization of WNV circulating along the Kenyan coast, revealing the co-detection of Lineage 1a and the first identification of Lineage 2 in this region. By combining field surveillance, whole-genome sequencing, evolutionary analyses, and lineage-specific replication assays across multiple vertebrate and mosquito cell lines, we demonstrate clear genetic and biological differences with implications for transmission and adaptation. Importantly, the detection of WNV in a male Aedes aegypti mosquito and recovery of full genomes offers compelling evidence of transovarial transmission, a mechanism that may support viral maintenance independent of vertebrate hosts. These findings expand current knowledge of WNV ecology in Africa and underscore the need for continued genomic surveillance to detect emerging variants and inform public health strategies.

genomics↗

Identification and genetic characterization of Jingmen tick virus from ticks sampled in select regions of Kenya; 2022-2024

Jingmen tick virus (JMTV), an emerging segmented RNA virus classified as an ungrouped flavivirus, poses a growing public health concern globally. Known for its association with febrile illnesses and wide host range, JMTV has been detected in Rhipicephalus, Hyalomma, and Amblyomma ticks collected from cattle, goats, sheep, camels, and chickens in pastoral regions of Kenya, including Baringo, Mandera, Malindi, Lamu, Mombasa, Wajir, Isiolo, and West Pokot. Using viral metagenomics next-generation sequencing, this study analysed adult ticks (n=1547, 72 pools). A total of 53% (38/72) pools were positive for at least one viral pathogen, with JMTV detected in 87% (33/38) of these pools across all study sites. Phylogenetic analyses revealed evidence of distinct Kenyan JMTV strains, with sequence segments from Malindi and Wajir clustering uniquely in their own clade; suggesting potential localised evolutionary pressures. Time calibrated phylogeny for the segment 1(RdRp) suggested varied ancestral origins and evolutionary relationships for the JMTV strains. MEME, BUSTED and FUBAR methods implemented in the Data-Monkey, unanimously identified codon 290 in segment 1 and 30 in segment 4 to be undergoing episodic positive selection. Recombination analysis performed using the RDP4 recombination detection tool indicated a recombination event in segment 2 of the Lamu JMTV strain that was confirmed by seven detection methods and visualised in BootScan. These findings suggest that Kenyan JMTV strains are undergoing positive selection, potentially driven by unique ecological and host factors. Segmented genome evidence of recombination highlights the increasing viruss potential for antigenic diversity. Host diversity and virus phylogenetic patterns underscore the zoonotic potential and its capacity for regional spread, emphasizing the critical need for enhanced vector surveillance. Temporal and ecological drivers like seasonal tick activity and livestock movement warrant investigation to elucidate JMTV transmission dynamics. Prioritizing tick-borne virus surveillance in Kenya will strengthen public health strategies and mitigates emerging viral risks.

evolutionary biology↗

Characterization of West Nile virus Koutango lineage from Phlebotomine Sandflies in Kenya 2021

The West Nile virus (WNV), primarily transmitted by mosquitoes, is one of the most widespread flaviviruses globally, with past outbreaks occurring in the USA and Europe. Recent studies in parts of Africa, including Kenya, have identified the West Nile virus Koutango lineage (WN-KOUTV) among phlebotomine sandfly populations, however, our understanding of this virus remains limited. Hence, this study aimed to characterize WN-KOUTV from phlebotomine sandflies. Sandflies were sampled between 12-16th March 2021 from six villages in Baringo South, Kenya, using CDC light traps. Female sandflies were taxonomically identified and pooled based on genus. Virus isolation was performed in Vero cells. Viral genome was determined using next-generation sequencing. Phylogenetic and molecular clock analyses were done to decipher the viruss evolutionary relationships. Comparative analyses of amino acid sequences were performed to determine variations. Protein modeling in Pymol was conducted to elucidate variations in key protein regions. Evolutionary pressure analysis investigated the selection pressures on the virus. In vitro experiments were done to investigate the virus growth kinetics in mammalian (Vero-E6) and mosquito (C636) cells. We report the isolation of WN-KOUTV from Salabani Baringo South, Kenya. The isolated WN-KOUTV clustered with previously identified WN-KOUTV strains. Comparative analysis revealed unique amino acid at NS5 653. Diversifying pressure was acting NS3 267 of the WN-KOUTV lineage. WN-KOUTV replicates efficiently in Vero-E6 and C636 cells comparable to West Nile virus Lineage 1a, isolated from mosquitoes. The isolation of WN-KOUTV in sandflies points to them as potential vectors, however, vector competence studies would confirm this. The efficient replication in mammalian and mosquito cell lines elucidated its adaptability to host and vector. We speculate the close genetic relationship of WN-KOUTV strains is enabled by the bird migratory route between East and West Africa. If proven, this may point to a potential future pandemic pathway for this virus.

microbiology↗

Detection of pathogenic bacteria in ticks from Isiolo and Kwale counties of Kenya using metagenomics.

Ticks are arachnid ectoparasites which rank second only to mosquitoes in the transmission of human diseases including bacteria responsible for anaplasmosis, ehrlichiosis, spotted fevers, and Lyme disease among other febrile illnesses. Due to paucity of data on bacteria transmitted by ticks in Kenya, this study undertook a bacterial metagenomic-based characterization of ticks collected from Isiolo, a semi-arid pastoralist County in Eastern Kenya, and Kwale, a coastal County with monsoon climate on the southern Kenyan border with Tanzania. A total of 2,918 ticks belonging to 3 genera and 10 species were pooled and screened in this study. Tick identification was confirmed through the sequencing of Cytochrome C Oxidase Subunit 1 (COI) gene. Bacterial 16S rRNA gene PCR amplicons obtained from the above samples were sequenced using the MinION (Oxford Nanopore Technologies) platform. The resulting reads were demultiplexed in Porechop, followed by trimming and filtering in Trimmomatic before clustering using Qiime2-VSearch. A SILVA database pretrained naive Bayes classifier was used to taxonomically classify the Operational Taxonomic Units (OTUs). The bacteria of clinical interest detected in pooled tick assays were as follows: Rickettsia spp. 59.43% of pools, Coxiella burnetii 37.88%, Proteus mirabilis 5.08%, Cutibacterium acnes 6.08% and Corynebacterium ulcerans 2.43%. These bacteria are responsible for spotted fevers, query fever (Q-fever), urinary tract infections, skin and soft tissue infections, eye infections and diphtheria-like infections in humans, respectively. P. mirabilis, C. acnes and C. ulcerans were detected only in Isiolo. Additionally, COI sequences allowed for the identification of Rickettsia and Coxiella species to strain level in some of the pools. Diversity analysis revealed that the tick genera had high levels of Alpha diversity but the differences between the microbiomes of the three tick genera studied were not significant. The detection of Cutibacterium acnes, commonly associated with human skin flora suggests that the ticks may have contact with humans potentially exposing them to bacterial infections. The findings in this study highlight the need for further investigation into the viability of these bacteria and the competency of ticks to transmit them. Clinicians in these high-risk areas also need to be appraised for them to include Rickettsial diseases and Q-fever as part of their differential diagnosis.

bioinformatics↗