Overcoming barriers to single-cell RNA sequencing adoption in low- and middle-income countries.

The advent of single-cell resolution sequencing and spatial transcriptomics has enabled the delivery of cellular and molecular atlases of tissues and organs, providing new insights into tissue health and disease. However, if the full potential of these technologies is to be equitably realised, ancestrally inclusivity is paramount. Such a goal requires greater inclusion of both researchers and donors in low- and middle-income countries (LMICs). In this perspective, we describe the current landscape of ancestral inclusivity in genomic and single-cell transcriptomic studies. We discuss the collaborative efforts needed to scale the barriers to establishing, expanding, and adopting single-cell sequencing research in LMICs and to enable globally impactful outcomes of these technologies.

Isolation and characterization of biosurfactant-producing microbes isolated from the gastrointestinal system of broiler birds fed a commercial diet.

Antimicrobial drug resistance (AMR) from improper use of antibiotics in various livestock products is a growing hazard for humans worldwide, with current death rate in excess of 700,000 per annum linked to the problem. Microorganisms are a rich source of structurally distinct bioactive compounds designed to protect the microbes and can offset AMR challenge. A study was conducted at Chinhoyi University of Technology to isolate, identify and characterize biosurfactant secreting microbes from broiler bird's gastrointestinal tract. Analysis of variance was performed in Genstat software. 16S rRNA technique was used to identify the DNA of isolates, annotated by similarity using BLASTn analysis against the NCBI nucleotide database. Phylogenetic analysis was performed on the BLASTn outcome to have an appreciation of the evolutionary genetic relationships. Small intestine-derived samples had a wider hemolytic activity of 5.6 mm, with a 39% emulsification index. At 98.29% sequence similarity, the bacterium producing biosurfactants was identified as an Escherichia coli strain similar to the 7.1994/NIST 0056 strain. The biosurfactant substance is a derivative of decane with beta lactams, tetracyclines and sulfa drugs properties which were responsible for the observed antibacterial activity. We recommend endogenous biosurfactant production optimization experiments and in-vivo trials to evaluate the potential impacts of a biosurfactant based feed additive in broilers.

Identification of Zimbabwe's locally grown banana (Musa Spp.) cultivars using morphology and genome-targeted sequencing.

BACKGROUND: Banana production is increasingly under threat due to harsh weather conditions as a result of climate change and different diseases. As such there is a need for the preservation and the characterization of the banana cultivar population for the purposes of crop improvement. The identification of collected banana germplasm in Zimbabwe was conducted based on the Inter-transcribed spacer region as well as morphology. The study was conducted with the aim of distinguishing one cultivar from another towards genetic conservation as well as banana improvement. RESULTS: ITS 1 and ITS 4 region targeting primers were used to amplify the DNA from twelve cultivars as well as sequence. Blast results identified five Musa groups which are Musa balbisiana (BB), Musa ABB, Musa AB hybrid, Musa acuminata (AAA), and Musa acuminata subsp. Malaccensis (AA). Phylogenetic analysis was done on the sequences under study and a maximum likelihood tree was generated to determine relationships between the sequences. Further identification was done using the inflorescence, bract, and male bud and fruit characteristics of each cultivar complementing the molecular evaluation. CONCLUSION: Genetic and morphological identification of locally grown bananas was therefore successful. An important step towards identifying pure lines suitable for breeding.

Antibacterial activity of medicinal plants on the management of mastitis in dairy cows: A systematic review.

BACKGROUND: Mastitis is a disease of economic importance in dairy production systems. The common management regime for mastitis is the use of synthetic antibiotics, giving a new problem of antibiotic resistance. There is, therefore, a need to prospect for alternatives to conventional antibiotics from herbal plants. OBJECTIVES: This systematic review evaluates the use of plants as alternatives for the control of mastitis in dairy cattle, focussing on the effectiveness of studied plants and plant-based products and possible implications on the use of these products in livestock health. METHODOLOGY: The PRISMA model was implemented with searches done in five electronic databases: Scopus, ScienceDirect, PubMed, Ovid and Research4Life. Data were extracted from 45 studies with 112 plant species from plant species belonging to 42 different families. The specific keywords were 'mastitis', 'dairy cows' and 'medicinal plants'. RESULTS: The most cited plant species included Allium sativum L., Azadirachta indica and Eucalyptus globulus Labill with the latter further exploring its components. Microbial species causing mastitis mainly were Staphylococcus aureus and Escherichia coli. The extraction methods used included maceration approach using ethanol, methanol and water as solvents for phytochemicals and chromatographic techniques for essential oils. A few studies explored the mode of action, and toxicities of the herbal extracts as well as evaluating their efficacy in clinical trials using animal models. CONCLUSION: Plants with defined levels of phytochemicals were essential sources of antibacterials. Standardisation of analytical methods is required.

Warfarin pharmacogenetics in a black Zimbabwean cohort: an observational prospective study.

Aim: A prospective observational study was conducted to evaluate the feasibility of implementing clinical guidelines for warfarin dosing in black Zimbabwean patients. Methods: CYP2C9*5, CYP2C9*6, CYP2C9*8 and CYP2C9*11 and VKORC1 c. 1639 G>A variations were observed in 62 study patients. Results & Conclusion: Overall, 39/62 (62.90%) participants did not receive a warfarin starting dose as would have been recommended by Clinical Pharmacogenetics Implementation Consortium guidelines. US FDA and Dutch Pharmacogenetics Working Group guidelines are based on CYP2C9*2 and CYP2C9*3 only, hence, unlikely useful in this cohort, where such variants were not detected. Clinical Pharmacogenetics Implementation Consortium guidelines, on the other hand, have a specific recommendation on the African-specific variants CYP2C9*5, CYP2C9*6 and CYP2C9*11, and are hence suitable for implementation in Zimbabwe and would help optimize warfarin doses in patients in the study cohort.

Pharmacogenetics and pharmacokinetics of tamoxifen in a Zimbabwean breast cancer cohort.

Tamoxifen is the most used hormonal therapy for oestrogen receptor-positive breast cancer. CYP2D6 is the main enzyme in the metabolic pathway of tamoxifen to endoxifen. Variations in endoxifen plasma concentrations are associated with CYP2D6 polymorphisms. This study aimed to determine the association between the CYP2D6 polymorphisms and endoxifen plasma concentrations in a cohort of Zimbabwean breast cancer patients (n = 40). TaqMan genotyping and copy number assays were done to determine CYP2D6 genotypes. Tamoxifen and metabolites were quantitated using LC-MS/MS. The population had high frequencies of the CYP2D6 reduced function alleles, *17 (15%) and *29 (18%). The median endoxifen concentration was 4.78 ng/mL, and in 55% of the patients, mostly intermediate metabolizers were below the endoxifen therapeutic threshold of 5.97 ng/mL. The CYP2D6 phenotypes and activity scores were significantly associated with endoxifen plasma concentrations (P = 0.0151) and with endoxifen to N-desmethyl-tamoxifen ratios (P = 0.0006).

Pharmacogenetics of 6-mercaptopurine in a black Zimbabwean cohort treated for acute lymphoblastic leukaemia.

Background: 6-mercaptopurine usage is associated with myelotoxicity and increased risk in patients carrying metabolism-related genetic variations. This study aimed to determine the frequency of candidate gene polymorphisms and their association with 6-mercaptopurine intolerance. Methods: A total of 41 patients on acute lymphoblastic leukaemia treatment were genotyped for TPMT and NUDT15 (rs116855232) alleles, and their association with dose intensity was analyzed. Results: The defective TPMT*3C allele frequency was 9.8%. The median maintenance dose intensity for TPMT*1/*3C participants was considerably lower (47%) when compared with the TPMT*1/*1 wild-type (77%), although not statistically significant. Conclusion: This is the first pharmacogenetics study carried out in a black Zimbabwean leukemia patient cohort. The high defective TPMT*3C (9.8%) allele frequency points to the potential utility of pharmacogenetics testing for safe usage of 6-mercaptopurine in this population.

Acute lymphoblastic leukemia (ALL) is the most common malignancy affecting children in Zimbabwe and 6-mercaptopurine is frequently used as part of its treatment. However, 6-mercaptopurine is associated with side-effects such as severe neutropenia (a condition where you have a low number of white blood cells called neutrophils in your blood), with increased risk observed in patients carrying variants in genes involved in the metabolism of 6-mercaptopurine. Therefore, this study aimed to determine the frequency of polymorphisms in specific genes as well as their association with drug intolerance. A total of 41 patients on ALL treatment were studied. Review of treatment records was done to determine the cumulative 6-mercaptopurine dose and calculate dose intensity. Genotyping (to determine the versions of a gene a patient carries) for TPMT and NUDT15 (rs116855232) was performed and results correlated with drug dose intensity. The most frequent genotype was TPMT*1/*1, occurring in 80% of the participants. The remaining 20% were carriers with two different copies of TPMT (*1/*3C). The defective TPMT*3C variation occurred at 9.8% and none had TPMT*2, *3A, *3B or NUDT15 rs116855232 variants. Comparison analysis with dose intensity was done for 23 participants (56%) who had maintenance records available. The median dose intensity of 47% for TPMT*1/*3C participants was considerably low when compared to that of a normal TPMT*1/*1 patient, which was 77%. However, no statistically significant difference was observed between TPMT genotype and dose intensity. This is the first study in a group of leukemic Zimbabweans to investigate the frequency of TPMT and NUDT15 variants. With a high variation frequency of 9.8% for the defective TPMT*3C, pharmacogenetics testing for TPMT before treatment with 6-MP is recommended in the Zimbabwean population.

Pharmacogenetics of pain management in Zimbabwean patients with sickle cell disease.

Background: Pain is a common cause of hospitalization in sickle cell disease (SCD) patients. Failure to effectively control pain remains a challenge in patient care. Materials & methods: The authors conducted a cross-sectional study to determine the effect of CYP2D6 and UGT2B7 polymorphisms on pain management in 106 Zimbabwean SCD patients. Participant information was collected on a questionnaire. Genotyping was conducted using the GenoPharm® pharmacogenomics open array panel containing CYP2D6 and UGT genetic variants implicated in opioid response. Results: The reduced function alleles CYP2D6*17 and *29 had high frequencies of 15.9% and 12.9%, respectively. UGT2B7 rs73823859 showed a statistically significant correlation with pain levels (p = 0.0454). Conclusion: This study demonstrated the role of UGT2B7 polymorphism in SCD patient pain management.

Cardiotoxicity and pharmacogenetics of doxorubicin in black Zimbabwean breast cancer patients.

AIMS: Doxorubicin-induced cardiotoxicity (DIC) is a significant cause of mortality in cancer care. This study was conducted to establish the frequency of DIC in Zimbabwean breast cancer patients on doxorubicin and to test the DIC predictive power of genetic biomarkers. METHODS: A cohort of 50 Zimbabwean breast cancer patients treated with doxorubicin were followed up for 12 months with serial echocardiography and genotyped for UGTA1A6*4, SLC28A3 and RARG. Eleven per cent of the patients experienced DIC. RESULTS: The frequencies of SLC28A3 (rs7853758), UGT1A6*4 (rs17863783) and RARG (rs2229774) were 60.7%, 17.9% and 14.3%, respectively. No association between DIC and the three variants was observed. CONCLUSIONS: This is the first study on the prevalence of DIC and associated genetic biomarker predictive evaluation in Zimbabwean breast cancer patients. The genetic frequencies observed in our study were different to those reported in other populations. A larger sample size with a longer follow-up time will be necessary in future studies.

Purification and Amplification of DNA from Cellulolytic Bacteria: Application for Biogas Production from Crop Residues.

Polymerase chain reaction (PCR) is a popular molecular tool for detection of bacteria. PCR allows millions of copies of a target segment of DNA to be produced. The DNA is extracted from overnight grown cultures of pure bacterial isolates using either the organo-solvent method or a commercial DNA extraction kit. The quality and purity of the DNA is determined by performing gel electrophoresis on 0.8% agarose gel. The DNA is amplified by performing PCR assay. Bands of approximately 1.5 kb in size are obtained from the amplified products of DNA. The PCR products run on 1.5% agarose gel are visualized with UV light and imaged by gel documentation system. This chapter outlines the protocol for isolation and amplification of DNA from cellulolytic bacteria. Cellulolytic bacteria are considered a potential source of cellulases for pretreatment of crop residues during biogas production. PCR is considered a very powerful, sensitive, specific, fast, and reliable tool in molecular detection and diagnostics.

Phenotypic and genetic characterisation revealed the existence of several biotypes within the Neorautanenia brachypus (Harms) C.A. wild accessions in South East Lowveld, Zimbabwe.

BACKGROUND: Local communities in the South Eastern Lowveld of Zimbabwe have adopted the feeding of livestock with Neorautanenia brachypus (Harms) C.A. tuber to mitigate against climate change. Differences within Neorautanenia brachypus (Harms) tuber flesh colour and preferences by cattle have been observed, suggesting possible diversity within the N. brachypus plant community. This study aimed at distinguishing the N. brachypus wild plant species through phenotypic and genetic characterization using morphological descriptors and random amplified polymorphic (RAPD) markers respectively. Leaf samples were selected using judgmental sampling techniques from wards 11-15 in Sengwe (Chiredzi district) for leaf morphology and molecular characterization. RAPD-PCR analysis was done using 18-screened random decamer primers to confirm the diversity in the plant population. The similarity of the biotypes was evaluated using binary coding on the basis of the presence or absence of a morphological indicator as well as distinct DNA amplicon fragments. Primer 7.0.13 was used to estimate morphological and genetic similarities using the unweighted pair group method with arithmetic average (UPGMA). The cluster number was estimated using the Elbow method part of the R package. RESULTS: Initially, 14 biotype groups were identified from 96 accessions visually characterized basing of leaf characteristics. All the leaf biotypes displayed arcuate venation with differences observed for leaf shape, tip shape and leaf margins. The 14 biotypes clustered into six groups based on the binary data of the morphological characteristics. RAPD primers generated three hundred and sixty eight distinct amplicons with 77.5% being polymorphic from the 14 biotypes. The number of bands produced per primer ranged from four (OPF-02) to 44 (UBC-746). The PIC value ranged from 0.1327 to 0.1873 for the RAPD primers. Use of molecular markers collapsed the biotypes into five clusters. Both the leaf descriptors and RAPD markers showed the existence of genetic diversity within the wild accessions of N. brachypus. CONCLUSIONS: A combination of morphological and RAPD markers effectively refined the resolution of the genetic diversity within the N. brachypus wild accessions to nine biotypes. These findings have indicated to the existence of more than one biotype of N. brachypus with potentially different properties. The favorable biotypes can further be promoted through incorporation in pastures as alternative feed or complementary feed to livestock. As such the output of this study will serve as a guide for N. brachypus germplasm management and improvement.