Prevalence and Molecular Characterisation of Extended-Spectrum Beta-Lactamase-Producing Shiga Toxin-Producing <i>Escherichia coli</i>, from Cattle Farm to Aquatic Environments

Extended-spectrum beta-lactamase (ESBL)-producing bacteria are a major problem for public health worldwide because of limited treatment options. Currently, only limited information is available on ESBL-producing Shiga toxin-producing Escherichia coli (STEC) in cattle farms and the surrounding aquatic environment. This study sought to track and characterise ESBL-producing STEC disseminating from a cattle farm into the water environment. Animal husbandry soil (HS), animal manure (AM), animal drinking water (ADW), and nearby river water (NRW) samples were collected from the cattle farm. Presumptive ESBL-producing STEC were isolated and identified using chromogenic media and mass spectrophotometry methods (MALDI-TOF-MS), respectively. The isolates were subjected to molecular analysis, and all confirmed ESBL-producing STEC isolates were serotyped for their O serogroups and assessed for antibiotic resistance genes (ARGs) and for the presence of selected virulence factors (VFs). A phylogenetic tree based on the multilocus sequences was constructed to determine the relatedness among isolates of ESBL-producing STEC. The highest prevalence of ESBL-producing STEC of 83.33% was observed in HS, followed by ADW with 75%, NRW with 68.75%, and the lowest was observed in AM with 64.58%. Out of 40 randomly selected isolates, 88% (n = 35) belonged to the serogroup O45 and 13% (n = 5) to the serogroup O145. The multilocus sequence typing (MLST) analysis revealed four different sequence types (STs), namely ST10, ST23, ST165, and ST117, and the predominant ST was found to be ST10. All 40 isolates carried sul1 (100%), while blaOXA, blaCTX-M, sul2, blaTEM, and qnrS genes were found in 98%, 93%, 90%, 83%, and 23% of the 40 isolates, respectively. For VFs, only stx2 was detected in ESBL-producing STEC isolates. The results of the present study indicated that a cattle environment is a potential reservoir of ESBL-producing STEC, which may disseminate into the aquatic environment through agricultural runoff, thus polluting water sources. Therefore, continual surveillance of ESBL-producing STEC non-O157 would be beneficial for controlling and preventing STEC-related illnesses originating from livestock environments

Human-Associated Methicillin-Resistant Staphylococcus aureus Clonal Complex 80 Isolated from Cattle and Aquatic Environments

Background: Human-associated methicillin-resistant Staphylococcus aureus (HA-MRSA) has mainly been reported in South African pig and chicken farms. The prevalence of antibiotic-resistant genes (ARGs), virulence factors (VFs), and multilocus sequence types (MLSTs) associated with HA-MRSA in cattle farms has not been reported. Consequently, this study characterised LA-MRSA and its spread from cattle farms into the environment. Method: Husbandry soil (HS), nearby river water (NRW), animal manure (AM) and animal drinking water (ADW) were collected on and around a cattle farm. Presumptive MRSA isolates were identified from these samples using CHROMagar media and genotyped as MRSA sequence types (STs), selected ARGs, and VFs, using polymerase chain reaction. An MLST-based dendrogram was generated to link the farm MRSA strains with those in a nearby river. Results: The prevalence of MRSA was 30.61% for HS, 28.57% for ADW, 22.44% for NRW, and 10.20% for AM. Isolates from HS harboured the highest number of resistant genes, with 100% for mecA, 91.66% for ermA, and 58.33% for blaZ. However, no ermC or tetM genes were detected. MRSA isolates from AM harboured the lowest number of resistant genes. Only sec and seq enterotoxins were found in all the assessed MRSA isolates. MRSA from the farm revealed six STs (ST80, ST728, ST1931, ST2030, ST3247, and ST5440); all of STs belonged to clonal complex 80 (CC80). An MLST-based dendrogram based on the concatenated sequences of MLST genes under the maximum likelihood criterion revealed four clades of amalgamated MRSA isolates from various livestock environmental matrices, including the NRW. Conclusion: The results suggest that livestock environmental matrices might be reservoirs of MRSA that could subsequently disseminate through runoff to pollute water resources. Therefore, continued surveillance of HA-MRSA in livestock environments is warranted