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This study determined the prevalence, risk factors, and molecular characteristics of Listeria species detected in beef and beef products sampled in Mpumalanga province, South Africa. Four hundred beef and beef products were collected from 30 retail outlets in three districts (Bronkhorstspruit, Emalahleni, and Middelburg) within the province. Standard bacteriological and polymerase chain reaction (PCR) assays were used in the study. The overall prevalence of L. monocytogenes and other Listeria spp. in the samples was 8.3% (33/400) and 30% (120/400) (p <.05), respectively. For the five variables investigated, statistically significant effects were evident only for the region (p <.001) and type of product (p <.0001) for L. monocytogenes, the type of outlet (p =.011) and the type of product (p <.0001) for Listeria spp. Of the 20 types of beef and beef products tested, 15 (75%) and 17 (85%) were contaminated by L. monocytogenes and Listeria spp., respectively (p =.429). Among the four categories of products tested, the prevalence of L. monocytogenes was 7.3% (8/109), 10.6% (11/104), 7.5% (8/106), and 7.4% (6/81) for raw beef, ready‐to‐eat (RTE) products, milled beef, and offal & organs, respectively (p >.799). Among the 33 L. monocytogenes isolates, PCR genoserogroup IIa (42.4%, 1/2a‐3a) was most frequently detected. All (100%) of the isolates carried one or more of the eight virulence‐associated genes assessed, with genes inlC and inlJ detected in all the isolates. The overall prevalence of L. monocytogenes (8.3%) and the high frequency of virulent serogroups of L. monocytogenes commonly associated with human listeriosis pose a food safety risk to consumers of beef and beef‐based products contaminated by L. monocytogenes. [ABSTRACT FROM AUTHOR]

In this study, Listeria isolates (214) were characterized as follows: L. innocua (77.10%), L. monocytogenes (11.21%), L. welshimeri (5.61%), L. grayi (1.40%), L. seeligeri (0.93%), and L. species (3.73%) that were not identified at the species level, from beef and beef based products from retail and farms in Mpumalanga and North West provinces of South Africa. MLVA was further used to type Listeria innocua isolates (165) and Listeria monocytogenes isolates (24). The L. monocytogenes isolates were also serogrouped using PCR. The MLVA protocol for L. monocytogenes typing included six tandem repeat primer sets, and the MLVA protocol for L. innocua included the use of three tandem repeats primer sets. The L. monocytogenes serogroups were determined as follows: 4b-4d-4e (IVb) (37.50%), 1/2a-3a (IIa) (29.16%), 1/2b-3b (IIb) (12.50%), 1/2c-3c (IIc) (8.33%), and IVb-1 (4.16%). MLVA could cluster isolates belonging to each specie, L. monocytogenes, and L. innocua isolates, into MLVA-related strains. There were 34 and 10 MLVA types obtained from the MLVA typing of L. innocua and L. monocytogenes, respectively. MLVA clustered the L. monocytogenes isolates irrespective of sample category, serogroups, and geographical origin. Similarly, the L. innocua isolates clustered irrespective of meat category and geographical origin. MLVA was able to cluster isolates based on MLVA relatedness. The clustering of isolates from farms and retailers indicates transmission of Listeria spp. MLVA is an affordable, simple, and discriminatory method that can be used routinely to type L. monocytogenes and L. innocua isolates. [ABSTRACT FROM AUTHOR]

Listeriosis is a foodborne disease caused by Listeria monocytogenes species and is known to cause severe complications, particularly in pregnant women, young children, the elderly, and immunocompromised individuals. The aim of this study was to investigate the presence of Listeria species in food and water using both biochemical and species-specific PCR analysis. L. monocytogenes isolates were further screened for the presence of various antibiotic resistance, virulence, and biofilm-forming determinants profiles using phenotypic and genotypic assays. A total of 207 samples (composed of meat, milk, vegetables, and water) were collected and analyzed for presence of L. monocytogenes using species specific PCR analysis. Out of 267 presumptive isolates, 53 (19.85%) were confirmed as the Listeria species, and these comprised 26 L. monocytogenes, 3 L. innocua, 2 L. welshimeri, and 1 L. thailandensis. The remaining 21 Listeria species were classified as uncultured Listeria, based on 16SrRNA sequence analysis results. A large proportion (76% to 100%) of the L. monocytogenes were resistant to erythromycin (76%), clindamycin (100%), gentamicin (100%), tetracycline (100%), novobiocin (100%), oxacillin (100%), nalidixic acid (100%), and kanamycin (100%). The isolates revealed various multi-drug resistant (MDR) phenotypes, with E-DA-GM-T-NO-OX-NA-K being the most predominant MDR phenotypes observed in the L. monocytogenes isolates. The virulence genes prfA, hlyA, actA, and plcB were detected in 100%, 68%, 56%, and 20% of the isolates, respectively. In addition, L. monocytogenes isolates were capable of forming strong biofilm at 4 °C (%) after 24 to 72 h incubation periods, moderate for 8% isolates at 48 h and 20% at 72 h (p < 0.05). Moreover, at 25 °C and 37 °C, small proportions of the isolates displayed moderate (8–20%) biofilm formation after 48 and 72 h incubation periods. Biofilm formation genes flaA and luxS were detected in 72% and 56% of the isolates, respectively. These findings suggest that proper hygiene measures must be enforced along the food chain to ensure food safety. [ABSTRACT FROM AUTHOR]

These cross-sectional studies reported the occurrence, genetic characteristics, and factors associated with the distribution of Listeria species on cattle farms and beef abattoirs in Gauteng Province, South Africa. A total of 328 samples (faeces, feeds, silage, and drinking water) were collected from 23 cattle farms (communal, cow-calf, and feedlot), and 262 samples (faeces, carcass swabs, and effluents) from 8 beef abattoirs (low throughput and high throughput) were processed using standard bacteriological and molecular methods to detect Listeria species. The factors associated with the prevalence of Listeria species were investigated, and multiplex polymerase chain reaction (mPCR) was used to determine Listeria species, the pathogenic serogroups, and the carriage of eight virulence-associated genes by Listeria monocytogenes. The overall prevalence of Listeria species in cattle farms was 14.6%, comprising Listeria innocua (11.3%), Listeria monocytogenes (3.4%), Listeria welshimeri (0.0%) compared with 11.1%, comprising Listeria innocua (5.7%), Listeria monocytogenes (4.6%), Listeria welshimeri (0.8%) for beef abattoirs. Of the three variables (area, type of farm/abattoir, and sample type) investigated, only the sample types at abattoirs had a significant (P < 0.001) effect on the prevalence of L. innocua and L. welshimeri. The frequency of distribution of the serogroups based on 11 L. monocytogenes isolated from farms was 72.7% and 27.3% for the serogroup 1/2a-3a and 4b-4d-4e, respectively, while for the 12 L. monocytogenes isolates recovered from abattoirs, it was 25%, 8.3%, 50% and 16.7% for the serogroup 1/2a-3a, 1/2b-3b, 1/2c-3c, and 4b-4d-4e respectively (P < 0.05). All (100%) isolates of L. monocytogenes from the farms and abattoirs were positive for seven virulence genes (hlyA, inlB, plcA, iap, inlA, inlC, and inlJ). The clinical and food safety significance of the findings cannot be ignored., (© 2024. The Author(s).)

Listeria monocytogenes is sporadically detected on a range of ready to eat fresh produce lines, such as spinach and rocket, and is a threat to public health. However, little is known about the diversity of L. monocytogenes present on fresh produce and their potential pathogenicity. In this work, fifteen Listeria monocytogenes isolates from the UK fresh produce supply chain were characterised using whole genome sequencing (WGS). Additionally, isolates were characterised based on their ability to form biofilm. Whole genome sequencing data was used to determine the sequence type of isolates based on multi-locus sequence typing (MLST), construct a core single nucleotide polymorphism (SNP) phylogeny and determine the presence of virulence and resistance associated genes. MLST revealed 9 distinct sequence types (STs) spanning 2 lineages (I & II) with one isolate belonging to the ST6 subtype, strains from which have been recently implicated in two large, food-associated L. monocytogenes outbreaks in South Africa and across Europe. Although most of the 15 isolates were different, comparison of core genome SNPs showed 4 pairs of 'indistinguishable' strains (<5 SNPs difference). Virulence profiling revealed that some isolates completely lacked the Listeria pathogenicity island-3 (LIPI-3) amongst other virulence factors. Investigation of the inlA gene showed that no strains in this study contained a premature stop codon (PMSC), an indicator of attenuated virulence. Assessment of biofilm production showed that isolates found in the fresh produce supply chain differ in their ability to form biofilm. This trait is considered important for L. monocytogenes to persist in environments associated with food production and processing. Overall the work indicates that a genetically diverse range of L. monocytogenes strains is present in the UK fresh produce supply chain and the virulence profiles found suggests that at least some of the strains are capable of causing human illness. Interestingly, the presence of some genetically indistinguishable isolates within the 15 isolates examined suggests that cross-contamination in the fresh produce environment does occur. These findings have useful implications in terms of food safety and for informing microbial surveillance programmes in the UK fresh produce supply chain. • The Lm population in the UK leafy produce supply chain is genetically diverse • Isolates are related to epidemic strains and possess the necessary genes for disease • Biofilm production differs amongst closely related isolates • WGS provides valuable information on isolates that is of interest to stakeholders [ABSTRACT FROM AUTHOR]

Background: Globally, bacterial contamination of platelets remains the major transfusion‐related infectious risk. Limited data are available from low‐middle income countries. The South African National Blood Service routinely tests a percentage of apheresis platelets for bacterial contamination. We report on contamination rates from January 2011 to December 2016. Methods: A retrospective analysis of the surveillance data from apheresis platelets was performed. Two to 4 ml of each apheresis product was collected aseptically and divided equally into an aerobic and anaerobic culture bottle using the BacT/ALERT (bioMerieux) or BACTEC™ (Beckton Dickinson) systems. Bacterial identification was done using the MicroScan (Beckman Coulter) system. Results: A total of 87 841 apheresis platelets were collected between 2011 and 2016 of which 18% were tested. The average positive culture rate was 2·7%, ranging between 0·6% (2011) and 5·22% (2015). Gram‐positive bacteria constituted 94% of all isolates (269/285), ten (4%) gram‐negative bacteria and 6 (2%) fungi were detected. The most common bacterial isolates were Cutibacterium acnes (35%), coagulase‐negative Staphylococci (26%), Bacillus spp. (8%), Corynebacteria spp. (8%). A total of nine highly pathogenic bacteria were isolated: two Acinetobacter spp, one each of Enterobacter spp., Serratia spp., Klebsiella spp., one Staphylococcus aureus and three Listeria spp. Average time to positivity for bacteria was 5·9 days. No cases of sepsis or death were reported during this time. Conclusion: Bacterial contamination rates, mostly due to gram‐positive bacteria, increased since 2011. Bacterial screening must be aligned with international recommendations, and novel technologies should be explored to reduce the residual risk. [ABSTRACT FROM AUTHOR]

Objective: To describe maternal risk factors, presentations, peripartum findings, and pregnancy outcomes in Listeria monocytogenes-infected women., Methods: A retrospective descriptive case review. The records of 51 pregnant women infected with listeriosis who delivered infants between February 1, 2016 and February 28, 2018 at three academic hospitals in Johannesburg, South Africa, were included. The diagnosis of listeriosis was made on maternal/neonatal-sampled blood or tissue cultures., Results: Forty-eight (82.3%) Listeria infections of maternal and neonatal listeriosis were diagnosed on blood culture. The median gestational age at diagnosis was at a preterm gestation of 33 (20-43) weeks. Twenty-eight women (54.9%) had normal vaginal deliveries. Precipitous labor was described in 18 (39%) of these women. Fetal distress was the indication for cesarean section in 22 (41.2%) women. Meconium-stained amniotic fluid was found in 21 (61.7%) women at the time of delivery. The category of very low birth weight had 14 (27.4%) neonates with an Apgar score of less than 7 at 5 min. Maternal morbidities included chorioamnionitis (3 [5.8%]) and puerperal infections (7 [13.7%]). The HIV-positive anemic women had a tendency towards listerial infections., Conclusions: Symptoms of listeriosis were non-specific and diagnosis was detected on blood culture sampling. Risk factors included HIV seropositivity and were associated with puerperal infections and anemia., (© 2022 The Authors. International Journal of Gynecology & Obstetrics published by John Wiley & Sons Ltd on behalf of International Federation of Gynecology and Obstetrics.)

Abstract: In South Africa, the antimicrobial activity of many indigenous plants has been investigated. In general, studies have focused on planktonic bacteria, with less attention given to bacterial biofilms. Many organisms, however, including the opportunistic pathogen Listeria monocytogenes occur more frequently as biofilms. The aim of this study was to identify and select plants that exhibit the best antilisterial activity, isolate the bioactive compounds and determine their effect on the architecture of listerial biofilms. The ethyl acetate and chloroform extracts of thirteen plants were investigated for antilisterial activity. The ethyl acetate extract of Acacia karroo and Plectranthus ecklonii showed the best antilisterial activity, exhibiting a minimum inhibitory concentration (MIC) of 3.1mg/ml and 0.5mg/ml, respectively. These were further selected for the identification of bioactive compounds. Column chromatographic purification of the ethyl acetate extracts of the leaves of A. karroo led to the isolation of three known pure compounds, namely epicatechin (1), β-sitosterol (2) and epigallocatechin (3). Confocal scanning laser microscopy (CSLM) showed that the biomass of the listerial biofilm was reduced when the isolated compounds were added. The aggregation of cells that were exposed to β-sitosterol and epigallocatechin was reduced from 25μm as observed in untreated cells to <10μm in length. [Copyright &y& Elsevier]

Abstract  We assessed the prevalence of free-living and plankton-associated Listeria species in the final effluents of a South African wastewater treatment facility and its receiving watershed between August 2007 and July 2008 as well as the antibiotic susceptibilities of effluent isolates. The physicochemical quality of the raw sewage and treated effluents was also determined. Free-living Listeria were more prevalent (96%), compared to plankton-associated Listeria species (58–67%). Listeria pathogens were sensitive to 11 (55%) of the 20 tested antibiotics, and showed varying (7–71%) levels of resistance to eight antibiotics. Turbidity, COD, NO3, PO4 and Listeria density fell short of recommended standards after treatment; while pH, temperature, TDS, DO and NO2 were compliant with target quality after treatment. We conclude that final effluents of wastewater treatment plants are potential sources of Listeria pathogens in the aquatic milieu of South Africa. [ABSTRACT FROM AUTHOR]

Meat products have been implicated in many listeriosis outbreaks globally, however there is a dearth of information on the diversity of L. monocytogenes isolates circulating in food products in South Africa. The aim of this study was to investigate the population structure of L. monocytogenes isolated in the meat value chain within the South African market. Based on whole-genome sequence analysis, a total of 217 isolates were classified into two main lineage groupings namely lineages I (n = 97; 44.7%) and II (n = 120; 55.3%). The lineage groups were further differentiated into IIa (n = 95, 43.8%), IVb (n = 69, 31.8%), IIb (n = 28, 12.9%), and IIc (n = 25, 11.5%) sero-groups. The most abundant sequence types (STs) were ST204 (n = 32, 14.7%), ST2 (n = 30, 13.8%), ST1 (n = 25, 11.5%), ST9 (n = 24, 11.1%), and ST321 (n = 21, 9.7%). In addition, 14 clonal complex (CCs) were identified with over-representation of CC1, CC3, and CC121 in "Processed Meat-Beef", "RTE-Poultry", and "Raw-Lamb" meat categories, respectively. Listeria pathogenic islands were present in 7.4% (LIPI-1), 21.7% (LIPI-3), and 1.8% (LIPI-4) of the isolates. Mutation leading to premature stop codons was detected in inlA virulence genes across isolates identified as ST121 and ST321. The findings of this study demonstrated a high-level of genomic diversity among L. monocytogenes isolates recovered across the meat value chain control points in South Africa. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a highly fatal foodborne causative agent that has been implicated in numerous outbreaks and related deaths of listeriosis in the world. In this study, six L. monocytogenes isolated from ready-to-eat (RTE) meat products were analysed using Whole Genome Sequencing (WGS) to identify virulence and resistance genes, prophage sequences, PCR-serogroups, and sequence types (STs). The WGS identified four different STs (ST1, ST121, ST204, and ST876) that belonged to serogroup 4b (lineage I) and 1/2a (lineage II). Core genome, and average nucleotide identity (ANI) phylogenetic analyses showed that the majority of strains from serogroup 4b (lineage I) clustered together. However, two isolates that belong to serogroup 1/2a (lineage II) grouped far from each other and the other strains. Examination of reference-guided scaffolds for the presence of prophages using the PHAge Search Tool Enhanced Release (PHASTER) software identified 24 diverse prophages, which were either intact or incomplete/questionable. The National Center for Biotechnology Information- Nucleotide Basic Local Alignment Search Tool (NCBI-BLASTn) revealed that Listeria monocytogenes strains in this study shared some known major virulence genes that are encoded in Listeria pathogenicity islands 1 and 3. In general, the resistance profiles for all the isolates were similar and encoded for multidrug, heavy metal, antibiotic, and sanitizer resistance genes. All the isolates in this study possessed genes that code for resistance to common food processing antiseptics such as Benzalkonium chloride. [ABSTRACT FROM AUTHOR]

The article discusses efforts to control the outbreak of listeriosis, a foodborne disease caused by the bacterium, Listeria monocytogenes, in South Africa. Topics discussed include the high mortality rate of listeriosis among neonates, older people and immunocomprised adults, processed meat products as possible source of listeriosis, recall of contaminated products and translation of public communication materials in different languages and targeted groups at increased risk of listeriosis.

The prevalence of bacteria with multidrug-resistance (MDR) is a significant threat to public health globally. Listeria spp. are naturally ubiquitous, with L. monocytogenes particularly being ranked as important foodborne disease-causing microorganisms. This study aimed to evaluate the incidence and determine the antimicrobial resistance (AMR) profiles of multidrug-resistant Listeria spp. (MDRL) isolated from different environmental samples (river and irrigation water) in the Sarah Baartman District Municipality (SBDM), Eastern Cape Province (ECP), South Africa. Molecular identification and characterization were carried out using polymerase chain reaction (PCR) and isolates that exhibited phenotypic resistance were further screened for relevant antimicrobial-resistant genes (ARGs). Findings revealed a total of 124 presumptive Listeria isolates; 69 were molecularly confirmed Listeria species. Out of the confirmed species, 41 isolates (59%) were classified as L. monocytogenes while 9 (13%) were classified as L. welshimeri . All Listeria spp. exhibited phenotypic resistance against ampicillin, penicillin, and trimethoprim-sulphamethoxazole and further screening revealed ARGs in the following proportions: sulI (71%), bla TEM (66%), tetA (63%), and bla CIT (33%). Results confirmed the occurrence of ARGs among Listeria inhabiting surface waters of ECP. The present study indicates that the river water samples collected from SBDM are highly contaminated with MDRL, hence, constituting a potential health risk.