This study assessed the microbiological quality of various ready-to-eat foods sold in Alice, South Africa. Microbiological analysis was conducted on 252 samples which included vegetables, potatoes, rice, pies, beef and chicken stew. The isolates were identified using biochemical tests and the API 20E, API 20NE and API Listeria kits; results were analyzed using the one-way-ANOVA test. Bacterial growth was present in all the food types tested; high levels of total aerobic count were observed in vegetables, 6.8 ± 0.07 followed by rice, 6.7 ± 1.7 while pies had the lowest count (2.58 ± 0.24). Organisms isolated included: Listeria spp. (22%), Enterobacter spp. (18%), Aeromonas hydrophila (12%), Klebsiella oxytoca (8%), Proteus mirabilis (6.3%), Staphylococcus aureus (3.2%) and Pseudomonas luteola (2.4%). Interestingly, Salmonella spp. and Escherichia coli were not isolated in any of the samples. There was a statistically significant difference ( p < 0.05) in the prevalence of foodborne pathogens from hygienic and unhygienic cafeterias. The results indicated that most of the ready-to-eat food samples examined in this study did not meet bacteriological quality standards, therefore posing potential risks to consumers. This should draw the attention of the relevant authorities to ensure that hygienic standards are improved to curtain foodborne infections.

This study evaluated the effects of growth medium, temperature, and incubation time on biofilm formation by Enterobacter cloacae strains. The ability to adhere to a surface was demonstrated using a microtiter plate adherence assay whereas the role of cell surface properties in biofilm formation was assessed using the coaggregation and autoaggregation assays. The architecture of the biofilms was examined under scanning electron microscope (SEM). All the strains adhered to the well of the microtiter plate when incubated for 48 h, irrespective of the growth medium and incubation temperature. It was also noted that 90% and 73% of strains prepared from nutrient broth and cultured in brain heart infusion (BHI) broth and tryptic soy broth (TSB), respectively, were able to form biofilms, in contrast to 73% and 60% strains from nutrient agar and cultured in BHI and TSB respectively grown under similar conditions. However, no statistically significant difference was observed when the two methods were compared. The coaggregation index ranged from 12% to 74%, with the best coaggregate activity observed when partnered with Streptococcus pyogenes (54%-74%). The study indicates the suitability of BHI and TSB medium for the cultivation of E. cloacae biofilms, however, temperature and incubation time significantly affect biofilm formation by these bacteria.

Bovine tuberculosis (TB) caused by Mycobacterium bovis is a significant health threat to cattle and a zoonotic threat for humans in many developing countries. Rapid and accurate detection of M. bovis is fundamental for controlling the disease in animals and humans, and for the proper treatment of patients as one of the first-line anti-TB drug, pyrazinamide, is ineffective against M. bovis. Currently, there are no rapid, simplified and low-cost diagnostic methods that can be easily integrated for use in many developing countries. Here, we report the development of a loop-mediated isothermal amplification (LAMP) assay for specific identification of M. bovis by targeting the region of difference 4 (RD4), a 12.7 kb genomic region that is deleted solely in M. bovis. The assay's specificity was evaluated using 139 isolates comprising 65 M. bovis isolates, 40 M. tuberculosis isolates, seven M. tuberculosis complex reference strains, 22 non-tuberculous mycobacteria and five other bacteria. The established LAMP detected only M. bovis isolates as positive and no false positives were observed using the other mycobacteria and non-mycobacteria tested. Our LAMP assay detected as low as 10 copies of M. bovis genomic DNA within 40 minutes. The procedure of LAMP is simple with an incubation at a constant temperature. Results are observed with the naked eye by a color change, and there is no need for expensive equipment. The established LAMP can be used for the detection of M. bovis infections in cattle and humans in resource-limited areas.

The present study was carried out to investigate the potential of Listeria ivanovii isolates to exist as biofilm structures. The ability of Listeria ivanovii isolates to adhere to a surface was determined using a microtiter plate adherence assay whereas the role of cell surface properties in biofilm formation was assessed using the coaggregation and autoaggregation assays. Seven reference bacterial strains were used for the coaggregation assay. The degree of coaggregation and autoaggregation was determined. The architecture of the biofilms was examined under SEM. A total of 44 (88%) strains adhered to the wells of the microtiter plate while 6 (12%) did not adhere. The coaggregation index ranged from 12 to 77% while the autoaggregation index varied from 11 to 55%. The partner strains of S. aureus, S. pyogenes, P. shigelloides, and S. sonnei displayed coaggregation indices of 75% each, while S. Typhimurium, A. hydrophila, and P. aeruginosa registered coaggregation indices of 67%, 58%, and 50%, respectively. The ability of L. ivanovii isolates to form single and multispecies biofilms at 25°C is of great concern to the food industry where these organisms may adhere to kitchen utensils and other environments leading to cross-contamination of food processed in these areas.

This study assessed the antimicrobial susceptibility of 51 Listeria ivanovii and 33 Enterobacter cloacae strains isolated from various ready-to-eat foods sold in Alice, South Africa. Isolates were identified using standard microbiological tests and further confirmed using API 20E and API Listeria kits. The disc diffusion technique was used to screen for antimicrobial susceptibility against 15 antimicrobials; minimum inhibitory concentration of five antibiotics was determined by the broth dilution method. All the strains of E. cloacae (100%) and 96% of L. ivanovii isolates were resistant to at least four or more of the antibiotics; nineteen antibiotypes were obtained based on the antibiotics used in the study. Antibiotype A5: AR PGR VAR ER APR was predominant in both L. ivanovii (23.5%) and E. cloacae (57.5%) isolates. Marked susceptibility of Listeriaivanovii was observed against chloramphenicol, ciprofloxacin, streptomycin and trimethoprim/sulfamethoxazole (100%) each while E. cloacae registered 100% susceptibility to ciprofloxacin only. Various percentages of susceptibility was reported to chloramphenicol and gentamicin (91%) each, nalidixic acid (97%) and streptomycin (94%). The MIC90 ranged from 0.004–7.5 µg/mL with E. cloacae being the most susceptible organism. The study demonstrated the presence of multi-resistant strains of bacteria in ready-to-eat-foods and speculates that these foods could serve as important vehicles transmitting multi-resistant bacteria to humans.