Sourdough fermentation is a traditional biotechnological process used to improve the nutritional value, flavour profile, and textural attributes of baked goods. Sourdough principal microflora is represented by yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB). The yeasts are primarily responsible for leavening whereas the LABs and AABs carry out the acidification of the dough and both contribute to the flavour of the resulting bread. Our previous work explored the microbial species diversity of sourdoughs in Botswana and showed the unique microbial communities involved in sourdough production from different areas in Botswana. This study is aimed at characterizing yeasts and bacterial isolates from 9 traditional sourdoughs indigenous to Botswana. Here we report the functional characterization of 21 yeasts, 9 LAB, 4 AAB, and 11 other bacteria isolated from traditional sourdough as reported in our previous study. These isolates were characterized based on their ability to assimilate different carbon sources found in flour (maltose, glucose, sucrose, fructose, raffinose and maltotriose), capability to ferment carbon sources (maltose, glucose, sucrose and fructose) in flour and ability to withstand baking associated stresses (thermal stress, osmotic stress, oxidative stress and ethanol stress). This characterization aimed to compare them with conventional baker’s yeast used in modern bread making and to assess their potential for future commercialization. Our results show that the majority of isolated S. cerevisiae strains can utilise different carbon sources, ferment them and withstand baking associated stresses. These are key baking traits poorly expressed in the conventional baker’s yeast. In contrast, the isolated LAB and AAB exhibited limited carbon source utilization. Despite the noted poor utilization of these isolates, we noticed that L. plantarum (ME1-B1), B. cereus (GT1-B1) and B. zhangzhouensis were able to withstand baking associated stresses better than all other bacteria isolates. This study shows how isolated S. cerevisiae strains and non-conventional yeasts have comparable capability to the conventional baker’s yeast, highlighting their potential as sourdough starters and prospects for future commercialization.

Fungi are among the principal causes of deterioration and yield losses affecting maize farmers in Southern Africa, especially Botswana. Maize is one of Botswana's staple crops attacked by more than sixty diseases, and several species of insect pests with maize ear rot as the most damaging in the region. Among the field fungal potential pathogens, Aspergillus niger is one of the most predominant species causing maize ear rot. However, the genetic diversity and pathogenic potential of A. niger under field conditions of Botswana is still deficient. This study aimed to investigate the occurrence and genetic diversity of A. niger strains associated with maize ear rot in monoculture fields of southern Botswana (Ramotswa, Ramaphate and Kopong). The presence of A. niger was detected in 64 % of maize ear rot isolates sampled from the monoculture fields. Thin layer chromatography and ANPKS primer pair indicated 71.4 % of the strains to be potentially orchratoxigenic (i.e., positive test for Ochratoxin A). While internal transcribed spacer (ITS) gene sequences, assessed through UPGMA, principal component analysis (PCA) and Bayesian techniques revealed the Ramotswa population to be genetically divergent from Kopong and Ramaphatle populations. Our preliminary data suggests a possible infraspecific split of natural A. niger populations in Botswana, with probable implications for pathogen control related traits such as susceptibility to antifungal agents. This study also observed the occurrence of A. niger putative hybrids which could be detected with machine learning algorithms like Kernel-SVM (Kernel - Support Vector Model) as the main classifier, while Grid search and XGBoost (eXtreme gradient boosting) can be used to search for the optimal model. Since ochratoxin A poses significant food safety and human health issues, this study provided valuable insight into potential control of maize ear rot in Botswana. Understanding this genetic variation is crucial for developing effective strategies to control fungal outbreaks and reduce the risk of ochratoxin contamination, ultimately helping to protect both food security and public health.

Environmental pollution by hard-to-degrade polymers is on a steep rise and impacting the entire ecosystem. Microbial keratinases are pivotal in the breakdown of protein polymers that are otherwise resistant to most proteases. In this study, we isolated and identified bacteria from Sua pan soil through morphological, biochemical, and 16SrRNA sequencing approaches and further assessed these isolates for their keratinase production potential. Among the screened isolates, four bacteria, Bacillus cereus Bac 2, Bacillus sp. Bac 1, Pseudomonas aeruginosa Bac 3, and Achromobacter insuavis Bac 4, exhibited the highest degradation activity. B. cereus Bac 2 produced the widest clearance zone, whereas A. insuavis Bac 4 produced the narrowest clearance zone on feather meal agar. Protein bands observed in SDS-PAGE gels for the selected isolates further supported the presence of keratinolytic enzymes. We also investigated the effect of temperature and pH on keratinase activity and determined that all the keratinases were alkaline proteases, with B. cereus Bac 2, P. aeruginosa Bac 3, and Bacillus sp. Bac 1 showing maximum activity at a pH of 8.5, while A. insuavis Bac 4 had maximal activity at a pH of 8.0. Overall, our results indicated that B. cereus Bac 2 keratinase had significantly higher activity across all temperature and pH ranges investigated, compared to all the other isolates (p ≤ 0.0001). These findings highlight the potential application of bacterial isolates from alkalophilic environments, in the eco-friendly degradation of feather waste, as valuable by-products such as organic fertilizers, peptides, and amino acids.

Sourdough is one of the oldest technologies employed by humans to leaven bread because of its ability to enhance the flavour and structure of bread using micro-organisms. However, there is a lack of comprehensive information in Botswana regarding the diversity of sourdough starters and the fermentative micro-organisms responsible for spontaneous fermentation. The present study aimed to explore the microbial species diversity of sourdoughs in Botswana and gain insight into the unique microbial communities involved in sourdough production. A total of nine samples were collected from different areas in Botswana. The microbial diversity in sourdoughs was characterized through the sequencing of amplicons of the 16S ribosomal DNA and internal transcribed spacer regions. In silico polymerase chain reaction–restriction fragment length polymorphism and phylogenetics were utilized to determine the genetic diversity among the isolates. The dominant yeast species identified were Saccharomyces cerevisiae, Wickerhamomyces anomamlus, Pichia kudriazverii and kazachstania humilis. Additionally, the presence of Lactiplantibacillus plantarum, Lacticaseibacillus paracasei, Liquorilactobacillus nageli and Bacillus cereus was also detected. It is worth noting that two species of acetic acid bacteria (AAB), namely Acetobacter pasteurianus and A. indonesiensis, were isolated, though in low levels, but the finding is significant in sourdough fermentation. The low occurrence of AAB (acetic acid bacteria) species observed in this study could be an important finding, as these bacteria are considered understudied, yet they are known to contribute significantly to the final product.

Background Stress-tolerant yeasts are highly desirable for cost-effective bioprocessing. Several strategies have been documented to develop robust yeasts, such as genetic and metabolic engineering, artificial selection, and natural selection strategies, among others. However, the significant drawbacks of such techniques have motivated the exploration of naturally occurring stress-tolerant yeasts. We previously explored the biodiversity of non-conventional dung beetle-associated yeasts from extremophilic and pristine environments in Botswana (Nwaefuna AE et.al., Yeast, 2023). Here, we assessed their tolerance to industrially relevant stressors individually, such as elevated concentrations of osmolytes, organic acids, ethanol, and oxidizing agents, as well as elevated temperatures. Results Our findings suggest that these dung beetle-associated yeasts tolerate various stresses comparable to those of the robust bioethanol yeast strain, Saccharomyces cerevisiae (Ethanol Red™). Fifty-six percent of the yeast isolates were tolerant of temperatures up to 42 °C, 12.4% of them could tolerate ethanol concentrations up to 9% (v/v), 43.2% of them were tolerant to formic acid concentrations up to 20 mM, 22.7% were tolerant to acetic acid concentrations up to 45 mM, 34.0% of them could tolerate hydrogen peroxide up to 7 mM, and 44.3% of the yeasts could tolerate osmotic stress up to 1.5 M. Conclusion The ability to tolerate multiple stresses is a desirable trait in the selection of novel production strains for diverse biotechnological applications, such as bioethanol production. Our study shows that the exploration of natural diversity in the search for stress-tolerant yeasts is an appealing approach for the development of robust yeasts.

The Combretum genus belongs to the family Combretaceae. It consists of about 370 species of herbs, shrubs, and trees widely used by African traditional healers due to their efficacy in controlling different maladies. Some of the species have gained recognition in the pharmacopeia as drugs and herbal teas, both within Africa and beyond. The phytochemical and biological study of the genus is gradually expanding and has generated several evidence-based platforms for further studies and the development of useful phytopharmaceuticals. This review critically assesses flavonoids, which are the second-largest phytochemical class of compounds in the genus. Different search engines, including Google Scholar, Google, SciFinder, PubMed, ScienceDirect, and printed books, were employed with different search terms. Trends, structural patterns, biological activities, and several limitations within the genus have been discovered for further research follow-ups regarding eight nine (89) flavonoids (flavanols, flavans, flavanones, chalcones, anthocyanins, and flavones) isolated from the genus between 1990 and May 2022. The isolated flavonoids exhibited antidiabetic, anthelmintic, antimicrobial, antioxidant, antimalarial, and anticancer activities. Particularly, phosphoenolpyruvate carboxykinase (PEPCK) gene inhibitor, (−)-epicatechin (27) from Kinkeliba; the antioxidant compounds, velutin (57), belamcanidin (58), and cirsilineol (59) from C. fragrans and the anticancer effects of 3-O-(-L-rhamnopyranosyl)-3′,4′,5′,5,7-pentahydroxyflavone (78), and myricetin-3-O-glucoside (77) from C. platypetalum, are significant discoveries for further investigation of their therapeutic mechanisms of action and other preclinical protocols. Based on the scientific data generated from the genus and the traditional uses, evidence-based development of possible natural drug candidates should form the focus of future investigations.

Emerging environmental pathogenic fungal infections, including cryptococcosis, continue to pose a significant threat to humans with compromised immunity and, to some extent, healthy ones. Cryptococcus neoformans was originally identified as the main etiological agent of human cryptococcosis, but recent studies have also identified the occurrence of opportunistic infections caused by Cryptococcus gattii . These two saprophytic facultative yeasts present a paradox as they can infect humans without requiring a host for replication or survival, a phenomenon termed readymade virulence. Many cryptococcal virulence traits appear to have dual effects that provide survival advantages in both animal hosts and the environment. Several molecular techniques have been developed to provide in-depth knowledge of these species complexes. This review will focus on the description of the Cryptococcus neoformans and Cryptococcus gattii ( CnCg ) species complexes and associated cryptococcal pathogenesis, ecological niches, and virulence factors employed by the pathogens to cause disease.

Synthetic chemists are increasingly exploring alternative routes to target products, considering factors like environmental threats, side product formation, cost, and efficiency. This is coupled with the health challenges that require urgent responses from new chemical entities. In this context, antimicrobial resistance has exacerbated global diseases, making them difficult to treat with prescribed drugs and causing a persistent public health crisis. These triggered our interest in the search for alternative antimicrobial agents, exploring different synthetic strategies for known bioactive natural compounds for further investigations. In this study, Lewis acid catalyst (FeCl3) and tetrahydrofuran as solvents in the one-pot synthesis of (neo)flavonoid derivatives from readily available phenols and ethyl phenylpropiolate were discovered to afford the coumarins in yields of 43–95%. The synthetic derivatives of 7-hydroxy-4-phenyl-2H-chromen-2-one and analogs obtained were biologically investigated for antimicrobial activity. The compounds showed moderate to poor antimicrobial activity. The effects of different substituents (-R) on the yield of the reaction are also discussed. Future studies should enhance the yield of derivative products and use different assay models for biological activity studies to fully explore the compounds` biorelevance.

Opportunistic infections due to Cryptococcus neoformans and C. gattii species complexes continue to rise unabated among HIV/AIDS patients, despite improved antifungal therapies. Here, we collected a total of 20 environmental and 25 presumptive clinical cryptococcal isolates from cerebrospinal fluid (CSF) samples of 175 patients enrolled in an ongoing clinical trial Ambition 1 Project (Botswana-Harvard Partnership). Identity confirmation of the isolates was done using MALDI-TOF MS and PCR. We describe the diversity of the isolates by PCR fingerprinting and sequencing (Oxford Nanopore Technology) of the intergenic spacer region. Mating types of the isolates were determined by amplification of the MAT locus. We report an unusual prevalence of 42.1% of C. neoformans x C. deneoformans hybrids Serotype AD (n = 16), followed by 39.5% of C. neoformans Serotype A (n = 15), 5.3% of C. deneoformans, Serotype D (n = 2), 7.9% of C. gattii (n = 3), and 5.3% of C. tetragattii (n = 2) in 38 representative isolates that have been characterized. Mating type-specific PCR performed on 38 representative environmental and clinical isolates revealed that 16 (42.1%) were MATa/MAT alpha hybrids, 17 (44.7%) were MAT alpha, and five (13.2%) possessed MATa mating type. We used conventional and NGS platforms to demonstrate a potential link between environmental and clinical isolates and lay a foundation to further describe mating patterns/history in Botswana.

Mopane tree (Colophospermum mopane) is one of the main ecological niches of Cryptococcus neoformans, an opportunistic fungal pathogen that causes cryptococcosis primarily on immunocompromised hosts after inhalation of basidiospores from the environment. Hence, we investigated the prevalence, and phenotypically (antifungal resistance and biofilm formation capacity) and genotypically (mating type and genetic structure) characterized C. neoformans isolated from C. mopane, Acacia tortilis, Adansonia digitata and Ziziphus mucronata in Botswana. We report 7.1% and 2.9% prevalence of C. neoformans in C. mopane and other trees, respectively. All tested C. neoformans isolates were determined to be non-WT to fluconazole. Most isolates (65%) of C. neoformans isolates were biofilm producers. Mating type determination revealed a higher proportion of the globally rare MATa allele (53%) and a single MATα/MATa hybrid. The observed genotypeswere VNI (71%), VNB (23%) and VNB/VNB hybrids (6%). Native trees other than C. mopane are alternative ecological niches of antifungal resistant C. neoformans, and this represents a serious public health concern,and this represents a serious public health concern, especially for high-risk populations. Prevalence of C. neoformans on native trees and the observed emergence of hybrids (evidence of sexual recombination) highlight the need for increased surveillance and risk assessment within a One Health paradigm.

Combretum species exhibit potent antibacterial and antifungal properties, but limited studies exist for most species. This study reports for the first time the in vitro antimicrobial and antibiofilm activities of Combretum platypetalum leaf, stem, and root. Fungi and bacterial strains implicated in the causes of some of the conditions to which C. platypetalum is traditionally applied were evaluated by Agar-well diffusion and micro-dilution methods, and their biofilm inhibition capacities were investigated. The highest zone of fungal inhibition of 22.70 ± 2.08 mm against C. albicans and the highest zone of bacterial inhibition of 21.70 ± 2.08 mm against S. aureus were observed. The minimum inhibitory concentration (MIC) ranged between 0.0156 mg/mL and 8.0000 mg/mL for fungi and 0.3906 mg/mL and 8.0000 mg/mL for bacteria at P = 0.0002. The best minimum fungicidal concentrations (MFCs), at P = 0.003, were observed at 0.0630 mg/mL for the leaf against C. glabrata and C. neoformans and 0.5 mg/mL for the root against C. neoformans, respectively. The highest biofilm inhibition of 40.61%, 36.04%, and 23.85% was observed in the stem, root, and leaf extracts against C. glabrata. Chloroform leaf extract reveals ?-sitosterol as the active antifungal and antibacterial agent. Computational antifungal activity and molecular docking analysis of the structure versus target (sterol 14?- demethylase) show a binding affinity for druggability. The leaves and roots are the most potent parts of the plant, with leaf extracts showing non-synergistic effects. Bioguided isolation is recommended to fully explore the species' evidence-based therapeutic potential.

Increasing concerns toward food safety and public health have rendered the use of synthetic chemicals in agricultural environments unacceptable. A shift toward biologically safe approaches has been considered a preferred strategy within the food handling chain and has received increasing attention over the past years in managing undesirable microbial growth. Although several studies have looked at the mode of action of most antagonists, the manipulation of microbial communities in food safety has not been fully explored. Very little is known about the effect of microbial diversity and composition in developing a healthy environmental approach for pathogen management in the farm to fork continuum. In view of the progress made in recent years in metagenomic technologies, information generated should be used to develop a dynamic approach that will consider a comprehensive approach involving environmentally friendly strategies in dealing with food losses caused by microbes to ensure food safety. Thus, this review includes information on the latest biocontrol applications to suppress undesirable microbial growth and extend fresh produce shelf life along the farm to fork continuum. The role of recent trends related to the potential of microbiomes in food safety and quality is further discussed. The use of physical treatments against pathogen growth is also highlighted.

The increasing occurrence of methicillin-resistant Staphylococcus aureus (MRSA) in the environment, food and healthcare systems is a global public health concern. MRSA is reported to cause food poisoning, osteomyelitis and pyogenic infections of the skin, and consequently has been categorized as a high-priority pathogen by the World Health Organization. Here, we determined the presence of MRSA in clinical (n=56), food (n=150) and housefly samples (n=970) collected from two hospitals in Botswana. Characterization based on phenotypic (antimicrobial resistance, biofilm production) and genotypic (antimicrobial resistance genes and integrons) profiles were performed on all isolates. Of the total samples tested, 64 were positive for MRSA following conventional culture methods and PCR amplification of the mecA and mecC genes for confirmation of presumptive MRSA isolates. The confirmed isolates included 71 % (95 % CI 83.2-59.6) from clinical, 9 % (95 % CI 14--4.8) from food, and 1 % (95 % CI 1.6-0.4) collected from housefly samples. In total 89 % (n=57) isolates in the current study showed a multidrug resistance phenotype, among these, resistance to β-lactams and glycoside antibiotic classes were predominant. Genotypic characterization showed the domination of the blatem gene (95 %) followed by fox (63 %) and tetO (19 %) whilst vanA was only reported in 13 % of the isolates. Integrons were detected in 50 % (32/64) of the total MRSA isolates, and we report a high prevalence of etd gene, detected in 67 % (43/64) of the isolates followed by eta 38 % (24/64) whilst tsst-1 (3%) was the least detected genetic determinant. The genes etb and PVL were not detected in a ll the tested MRSA isolates. We provide the first report on the prevalence of MRSA isolated from the clinical-food-vector nexus harbouring biofilm and blatem genes, and antibiotic resistance profiles in Botswana. These results are significant for risk-assessment analysis and the development of improved MRSA infection prevention and control strategies.

Nine morphologically distinct halophilic yeasts were isolated from Makgadikgadi and Sua pans, as pristine and extreme environments in Botswana. Screening for biosurfactant production showed that Rhodotorula mucilaginosa SP6 and Debaryomyces hansenii MK9 exhibited the highest biosurfactant activity using Xanthocercis zambesiaca seed powder as a novel and alternative inexpensive carbon substrate. Chemical characterization of the purified biosurfactants by Fourier Transform Infra-Red spectroscopy suggested that the biosurfactant from R. mucilaginosa SP6 was a rhamnolipid-type whereas the biosurfactant from D. hansenii MK9 was a sophorolipid-type. The two biosurfactants exhibited antimicrobial activities against eight pathogenic bacteria and fungal strains (Proteus vulgaris, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Micrococcus luteus, Cryptococcus neoformans, Candida albicans and Aspergilus niger). The sophorolopid-type biosurfactant was found to be the most potent among the antimicrobial drug resistant strains tested. The findings open up prospects for the development of environmentally friendly antimicrobial drugs that use an inexpensive source of carbon to reduce the costs associated with the production of biosurfactants.

Cercospora Leaf Spot (CLS) of Swiss chard (Beta vulgaris L. var. cicla) remains an important foliar disease worldwide, yet the causative agent Cercospora beticola genetic variability remains incompletely understood in Botswana. To assess the diversity of C. beticola from diseased Swiss chard in southern Botswana, 78 isolates from two farms were analyzed. C. beticola was isolated from Swiss chard at very high frequencies from both Bokaa (91%) and Glen Valley (86%) farms. However, statistical analyses indicated that the isolation rate of the pathogen was not affected by the sampling location (p=0.01, p>0.01). Phylogenetic analyses revealed that the 18 sequenced C. beticola isolates clustered into four major classes, which could not be differentiated by the sampling location. Similarly, genetic analysis revealed high genetic diversity of C. beticola strains from the two farms, accounted for by within population diversity (greatest pairwise distance=0.004). The results presented herein underscore the importance of assessment of genetic diversity of pathogens which may be important in targeted control and management of plant diseases.

Albizia anthelmintica is a medicinal plant belonging to the Fabaceae family. It is widely used by smallholder farmers and pastoralists to treat internal parasites in their livestock. This study aimed to determine the antibacterial and antioxidant potential of A. anthelmintica on pathogenic veterinary isolates. 100% hexane (He100), 100% chloroform (Ch100), 100% ethanol (E100), and 70% ethanol (E70) extracts of the roots and barks of A. anthelmintica were tested against four bacterial strains (Escherichia coli, Clostridium perfringens, Salmonella enterica serovar Typhimurium and Proteus mirabilis). Thin layer chromatography- 2, 2-diphenyl-1- picryl hydrazyl (TLC-DPPH) assay was used to examine antioxidant potential of extracts. Antimicrobial activity was determined using the disc diffusion method and minimum inhibiting concentrations (MICs) values were determined using the micro-titre broth-dilution method. At a concentration of 500 µg/ml, E70 roots extract showed the highest % DPPH inhibition of 66.9%. Among the bark extracts, the highest free radical scavenging activity was observed in E70 extracts with 58.9% DPPH inhibition. Phytochemical analysis of the plant extracts revealed the presence of compounds which are known to exhibit medicinal properties such as tannins, terpenoids, quinones, saponins and fatty acids phenols. E100 bark extracts contained most of these compounds except flavonoids. Only alkaloids were not detected in any of the roots or bark extracts. Ch100 bark extracts showed the highest antimicrobial activity and all bacterial isolates were resistant to the E100 root extracts. Ch100 root extracts showed the lowest minimum inhibition concentration of 0.625 mg/ml against S. enterica serovar Typhimurium. Findings of this study show that some of the root and bark extracts of the A. anthelmintica plant have both antimicrobial and antioxidant properties. These findings can possibly be relevant in the development of novel medication against veterinary pathogens. Furthermore, this study will guide similar studies. Key words: Antibacterial, antioxidant, Albizia anthelmintica, phytochemical, minimum inhibitory concentrations (MICs).

The powdery mildew of butternut squash ( Cucurbita moschata Duchesne ex Poiret) caused by Podosphaera xanthii (Castagne) U. Braun and Shiskoff is the most severe disease in Botswana. The disease causes a significant reduction in quality and quantity of butternut fruits. Greenhouse experiments were conducted during 2007 and 2008 at the Department of Agricultural Research Station, Sebele, Botswana to evaluate the efficacy of locally available protectant and systemic fungicides on the control of the powdery mildew of butternut squash. In 2007 treatment trial included a protectant fungicide, chlorothalonil and two systemic fungicides, benomyl and bupirimate which were used separately, and tank mixes of each of the systemic fungicide with the protectant fungicide. Fungicidal treatment in 2008 included an additional protectant fungicide, mancozeb which was used alone and also tank mixed with systemic fungicides used in 2007. Chlorothalonil and mancozeb when used alone were not effective in controlling the disease with the disease severity (DS) of 23.52% and 9.43% respectively over 31.45 % disease severity in non-treated plants. Out of the two systemic fungicides, bupirimate (1.21% DS) was more effective over benomyl (2.89% DS). The data on systemic - protectant fungicide treatment were not significantly different (P<0.05). Bupirimate when combined with protectant fungicide either chlorothalonil (0.46% DS) or mancozeb (0.34% DS), and tank mixes of benomyl with chlorothalonil (0.91% DS), and with mancozeb (0.34% DS) proved to be very effective in the control of powdery mildew of butternut squash as compared to all other fungicidal treatments.

Staphylococcus aureus is one of the causes of foodborne diseases worldwide. Staphylococcal food poisoning ensues after ingestion of contaminated food and results in symptoms of gastroenteritis such as vomiting, abdominal cramps and diarrhea. The present paper aims to isolate Staphylococcus aureus from foods sold by street vendors in Gaborone, Botswana, and to determine its enterotoxigenic potential and antibiotic resistance profile. One hundred eight food samples comprising starch, meat, salads and vegetables portions were collected from these vendors and tested for the presence of S. aureus. Identification of Staphylococcus aureus to the species level was performed using the Vitek 2 automated identification and susceptibility testing system (BioMerieux, Marcy-I’Etoile, France). Enterotoxins were detected by the Reversed Passive Latex Agglutination method (SET-RPLA). Results showed that 49 (45%) of the samples tested positive for Staphylococcus aureus . The organism was isolated at higher frequencies in vegetables and starchy foods (34.7%) than in meats (30.6%). These differences in isolation rates however, were not statistically significant (p> 0.05). Staphylococcus aureus isolates were found to be resistant to penicillin G (52.4%), tetracycline (38.1%), methicillin (26.2%) and vancomycin (11.9%). Four Staphylococcal enterotoxin types A-D, were detected among the isolates. Staphylococcal enterotoxin D was the most prevalent (52.9%), while enterotoxin C was produced by the least number of isolates (5.9%). Of note, five isolates simultaneously expressed two or more enterotoxin types in varying combinations. The present study underscores a potential risk of staphylococcal food poisoning and transmission of methicillin resistant S. aueus strains for consumers of street vended food products in Gaborone, Botswana especially in the absence of a quality assurance regulatory framework. As a mitigating factor, sensitization of street food vendors on the importance of food and personal hygiene is strongly recommended.