Biology subjects

Musila, L.

Publications and source records attributed to Musila, L..

3 recordsLinked to original sources

bioRxiv2023

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↗

bioRxiv2022

Antimicrobial resistance and virulence characteristics of Klebsiella pneumoniae isolates in Kenya

Klebsiella pneumoniae is a globally significant opportunistic pathogen causing healthcare-associated and community-acquired infections. This study examined the epidemiology and the distribution of resistance and virulence genes in clinical K. pneumoniae strains in Kenya. Eighty-nine K. pneumoniae isolates were collected over six years from five counties in Kenya and were analyzed using whole genome sequencing and bioinformatics. These isolates were obtained from community-acquired (62/89) and healthcare-associated infections (21/89), and the hospital environment (6/89). Genetic analysis revealed the presence of blaNDM-1 and blaOXA-181 carbapenemase genes and the armA and rmtF genes known to confer pan-aminoglycoside resistance. The most abundant extended-spectrum beta-lactamase genes identified were blaCTX-M-15 (36/89), blaTEM (35/89), and blaOXA (18/89). In addition, one isolate had a mobile colistin resistance gene (mcr-8). Fluoroquinolone resistance-conferring mutations in gyrA and parC genes were also observed. The most notable virulence factors were those associated with hyper-virulence (rmpA/A2 and magA), yersiniabactin (ybt), salmochelin (iro), and aerobactin (iuc and iutA). Thirty-eight distinct sequence types were identified, including known global lineages ST14, ST15, ST147, and ST307, and a regional clone ST17 implicated in regional outbreaks. In addition, this study genetically characterized two potential hypervirulent isolates and two community-acquired ST147 high-risk clones that contained carbapenemase genes, yersiniabactin, and other multidrug resistance genes. These results demonstrate that the resistome and virulome of Kenyan clinical and hospital environmental K. pneumoniae isolates are diverse. The reservoir of high risk-clones capable of spreading resistance and virulence factors have the potential to cause unmanageable infection outbreaks with high morbidity and mortality. AUTHOR SUMMARYKlebsiella pneumoniae is one of the human-disease-causing bacteria that easily acquires and spreads antibiotic resistance genes and is thus a serious threat to human health. We studied both the antibiotic resistance genes and the genes it uses to cause disease (virulence). Forty-two percent of our isolates were multidrug resistant (MDR). They carried several resistance and virulence genes bound in mobile circular DNA molecules called plasmids which easily migrate and spread the genes between bacteria. We identified 38 distinct K. pneumoniae strains (STs) distributed within the study sites. Fifteen isolates were classified under the groups of K. pneumoniae strains known to cause global infection outbreaks such as ST14, ST15 and ST147 collected from Nairobi and Kisumu, hotspot areas for spread of resistance. In particular, two ST147 isolates were resistant to carbapenems and one isolate to colistin, which are last line antibiotics. We also identified two isolates with the potential to cause high levels of disease. We concluded that the presence of highly resistant and virulent strains in the hospital and community demonstrates a need for the continuous monitoring and management of MDR K. pneumoniae infections to prevent disease outbreaks that are difficult to control and that lead to high death rate.

microbiology↗

bioRxiv2018

Strain typing and characterization of virulence genes in clinical Staphylococcus aureus isolates from Kenya

Staphylococcusaureus strain typing is an important surveillance tool as particular strains have been associated with virulence and community and hospital acquired MRSA outbreaks globally. This study sought to determine the circulating strain types of S.aureus in Kenya and establish the virulence genes among the strains. Clinical S.aureus isolates from 3 hospitals in Kenya were sequenced on the Illumina Miseq and genomes assembled and annotated on PATRIC. Results demonstrated great diversity among the isolates with identification of 6 distinct CC (8,22,15,80,121,152), 8 ST types (8, 15, 22,80,121,152,241, 1633) and 8 spa types (t005, t037, t064, t084, t233, t2029, t272,t355). Novel STs (4705, 4707) and a novel spa type (t17826) were identified. The most prominent clonal complex was CC 152 comprised of only MSSA. A majority of MRSA isolates (3/4) typed to ST 241, CC8. One MRSA isolate typed to a novel ST 4705. All isolates were screened for a panel of 56 known virulence genes (19 adhesins, 9 hemolysins, 5 immune evasion proteins, 6 exo-enzymes and 19 toxins). 9 toxin genes were detected among the isolates with CC8 isolates having the highest numbers of toxin genes. An MSSA isolate (CC8) from a severe burn infection had the highest number of toxin genes (5). All MRSA isolates (CC8) had only 2 toxins, SEK and SEQ, whereas a majority of the MSSA isolates either had 0 or [≥]2 toxins. SEK+SEQ and TSST-1+SEB+SEL toxin combinations were observed among patients whose disease resulted in hospitalization, an indicator of severe infections. This study confirms the highly heterogeneous S.aureus population in Kenya. MSSA appear to have the potential of accumulating more toxin genes than MRSA. This co-occurrence of major toxin genes, some associated with MRSA, highlights the potential risks of outbreaks of highly virulent MRSA infections which would pose treatment challenges.

microbiology↗