Prevalence Of Listeria Monocytogenes Research Articles

Prevalence of Listeria monocytogenes in Milk and Dairy Product Supply Chains: A Global Systematic Review and Meta-analysis.

Listeria monocytogenes, one of the main foodborne pathogens, is commonly found in milk and dairy products. This study aimed to estimate the presence of L. monocytogenes in milk and dairy product supply chains using a meta-analysis based on PubMed, Embase, Web of Science, and Scopus databases. A total of 173 studies were included in this meta-analysis. The pooled prevalence in the supply chain environment was 8.69% (95% confidence interval [CI]: 5.30%-12.78%), which was higher than that in dairy products (4.60%, 95% CI: 1.72%-8.60%) and milk products (2.93%, 95% CI: 2.14%-3.82%). Subgroup analysis showed that L. monocytogenes prevalence in raw milk (3.44%, 95% CI: 2.61%-4.28%) was significantly higher than in pasteurized milk (0.60%, 95% CI: 0.00%-2.06%). The highest prevalence of L. monocytogenes in milk and dairy products was observed in North America (5.27%, 95% CI: 2.19%-8.35%) and South America (13.54%, 95% CI: 3.71%-23.37%). In addition, studies using culture and molecular methods (5.17%, 95% CI: 2.29%-8.06%) had higher prevalence than other detection methods. Serogroup 1/2a and 3a (45.34%, 95% CI: 28.74%-62.37%), serogroup 1/2b and 3b (14.23%, 95% CI: 6.05%-24.24%), and serogroup 4b/4e (13.71%, 95% CI: 6.18%-22.83%) were dominant in these studies. The results of this study provide a better understanding of the prevalence of L. monocytogenes in milk and dairy product supply chains and suggest a potential foodborne pathogen burden.

Foodborne bacteria in milk and milk products along the water buffalo milk chain in Bangladesh

Controlling foodborne pathogens in buffalo milk is crucial for ensuring food safety. This study estimated the prevalence of nine target genes representing seven critical foodborne bacteria in milk and milk products, and identified factors associated with their presence in buffalo milk chain nodes in Bangladesh. One hundred and forty-three milk samples from bulk tank milk (n = 34), middlemen (n = 37), milk collection centers (n = 37), and milk product shops (n = 35) were collected and analyzed using RT-PCR. Escherichia (E.) coli, represented through yccT genes, was the most prevalent throughout the milk chain (81–97%). Chi-squared tests were performed to identify the potential risk factors associated with the presence of foodborne bacteria encoded for different genes. At the middleman level, the prevalence of E. coli was associated with the Mymensingh, Noakhali, and Bhola districts (P = 0.01). The prevalence of Listeria monocytogenes, represented through inlA genes, and Yersinia (Y.) enterocolitica, represented through yst genes, were the highest at the farm level (65–79%). The prevalence of both bacteria in bulk milk was associated with the Noakhali and Bhola districts (P < 0.05). The prevalence of Y. enterocolitica in bulk milk was also associated with late autumn and spring (P = 0.01) and was higher in buffalo-cow mixed milk than in pure buffalo milk at the milk collection center level (P < 0.01). The gene stx2 encoding for Shiga toxin-producing (STEC) E. coli was detected in 74% of the milk products. At the middleman level, the prevalence of STEC E. coli was associated with the use of cloths or tissues when drying milk containers (P = 0.01). Salmonella enterica, represented through the presence of invA gene, was most commonly detected (14%) at the milk collection center. The use of plastic milk containers was associated with a higher prevalence of Staphylococcus aureus, represented through htrA genes, at milk product shops (P < 0.05). These results suggest that raw milk consumers in Bangladesh are at risk if they purchase and consume unpasteurized milk.

Contamination of Food Products by Listeria monocytogenes in Iran during 2010-2023

Introduction: Mortality rate of Listeria infection is high and has been reported to be up to 30%. In this regard, the present study aimed to provide more information about food sources contaminated with the Listeria monocytogenes and offer suitable strategies to control this pathogen in supply centers, slaughterhouses, and processing centers. Method: Results of investigations on the prevalence of L. monocytogenes in food (seafood and aquatic products, red meat, poultry, and dairy products) during 2010-2023 were reviewed. In total, 61 acceptable studies were involved in the present research. After the selection of papers, their key results were extracted, including the prevalence of Listeria monocytogenes, cultivation method, sample type, and sampling location. Results: Listeria monocytogenes is prevalent in all types of seafood products, red meat, poultry, dairy products, and vegetables, as well as ready-to-eat and ready-to-cook foods. Consuming raw and undercooked foods contaminated with animal and human feces and direct contact with feces can cause Listeria infection in humans. L. monocytogenes is sensitive to amoxicillin, ampicillin, penicillin, and gentamicin antibiotics; therefore, these antibiotics can be used for listeriosis treatment. Conclusion: In previous studies, the importance of traditional meat and dairy supply centers in L. monocytogenes transmission has been addressed to a lesser extent. It can be concluded that appropriate standards should be developed and applied to control this food pathogen by regulatory systems. In addition, regular and periodic monitoring of food products and providing necessary training to people about how to prevent food poisoning can play an effective role in reducing listeriosis.

Investigation of the seasonal prevalence, phenotypic, and genotypic characteristics of Listeria monocytogenes in slaughterhouses in Burdur.

This study examined Listeria monocytogenes isolates from two slaughterhouses in Burdur province, southern Turkey, over four seasons for antibiotic resistance, serogroups, virulence genes, in vitro biofilm forming capacity, and genetic relatedness. Carcass (540) and environment-equipment surface (180) samples were collected from two slaughterhouses (S1, S2) for 1 year (4 samplings). Of the 89 (12.4%) positive isolates, 48 (53.9%) were from animal carcasses, and 41 (46.1%) from the environment-equipment surfaces. Autumn was the peak season for Listeria monocytogenes compared to summer and spring (P<0.05). In addition, the most common serotype between seasons was 1/2c. Except for plcA and luxS genes, all isolates (100%) harbored inlA, inlC, inlJ, hlyA, actA, iap, flaA genes. Listeria monocytogenes isolates were identified as belonging to IIc (1/2c-3c; 68.5%), IVb (4b-4d-4e; 29.2%), and IIa (1/2a-3a; 2.2%) in the screening using multiplex polymerase chain reaction-based serogrouping test. A total of 65 pulsotypes and 13 clusters with at least 80% homology were determined by using pulsed field gel electrophoresis on samples that had been digested with ApaI. Thirty-four (38.2%) of the isolates were not resistant to any of the 14 antibiotics tested. The antibiotic to which the isolates showed the most resistance was rifampicin (44.9%). Serotype 1/2c was the most resistant serotype to antibiotics. Despite having biofilm-associated genes (inlA, inlB, actA, flaA, and luxS), a minority (11%) of isolates formed weak biofilm. This study revealed seasonal changes prevalence of Listeria monocytogenes, particularly higher in autumn, posing a greater risk of meat contamination. Notably, Serotype 1/2c showed significant prevalence and antibiotic resistance. Indistinguishable isolates indicated cross-contamination, underscoring the importance of prioritized training for slaughterhouse personnel in sanitation and hygiene protocols.

A comprehensive systematic review and meta-analysis of Listeria monocytogenes prevalence in food products in South Korea

Listeria monocytogenes is a foodborne pathogen that can cause deadly severe listeriosis. While systematic review and meta-analysis are powerful tools for comprehensive analysis by pooling every related study, these approaches to L.monocytogenes contamination food have yet to be studied in South Korea. We aimed to identify high-risk L.monocytogenes foods in South Korea through a prevalence survey of retail food products for the first time. A total of 13,684 samples of 59 articles were used for meta-analysis through the systematic review, and the results were synthesized using a random-effects model considering the heterogeneity. The overall pooled prevalence was 2.26 % (95 % CI: 1.44–3.52 %). Among nine food categories, meat exhibited the highest prevalence at 8.32 % (95 % CI: 4.42–12.14 %) after sample size restriction. Specifically, a post-hoc sensitivity analysis was conducted to identify the prevalence difference among subgroups and the source of heterogeneity. Intriguingly, the analysis revealed chicken as the primary contributor to the elevated prevalence of L.monocytogenes, a key factor deriving the observed heterogeneity. This study carries significant implications for public health and food safety in Korea. Furthermore, knowledge of differences in prevalence levels in various foods will be able to be used as a predictive guideline for foodborne outbreaks.

Prevalence, identification and antimicrobial resistance of Listeria monocytogenes and Listeria spp. isolated from poultry and pork meat

SummaryThe aim of this work was to evaluate the prevalence of Listeria monocytogenes and other Listeria spp. in chicken, duck, quail, turkey and pork meat, including the antibiotic resistance of isolated strains. A total of 184 meat samples were collected from different retailers in La Rioja (Spain). The presence of Listeria spp. and L. monocytogenes were detected in 24.46% and 10.32% of the meat samples respectively. L. monocytogenes was the predominant Listeria spp. found in chicken, quail and pork meat, while L. innocua and L. welshimeri were the predominant species in duck and turkey meat respectively. A total of thirty‐three strains (55.93%) of Listeria spp. were found to be multi‐resistant (resistant to ≥3 families of antibiotics). The highest multi‐resistant rates were observed in L. monocytogenes (73.68%) and L. innocua (70.59%), followed by L. ivanovii (50%). Resistance to ampicillin and trimethoprim–sulfamethoxazole were found in L. monocytogenes strains isolated from chicken, being of special concern, since these antibiotics are used in the treatment of listeriosis. Special measures should be taken to reduce meat contamination such as adequate handling, correct preparation (cooking) and cleaning and disinfection in order to avoid cross‐contamination.

Prevalence of Listeria monocytogenes in dairy products in Turkey: A Meta-Analysis

This study was aimed to conduct a meta-analysis to determine the seroprevalence of Listeria monocytogenes in dairy products in Turkey. A systematic literature search was conducted using the Medline/PubMed, Science Direct, Web of Science, and Scopus databases to identify studies reporting the seroprevalence of L. monocytogenes in dairy products. The search was conducted using the following keywords: (“ice cream” or “curd” or “cream” or “cheese” or “butter” or “yoğurt” or “dairy” or “milk”) and (“Listeriosis” or “Listeria monocytogenes” or “Listeria”) and (“Turkiye” or “Turkey”) and (“seropositivity” or “seroprevalence” or “prevalence”). A comprehensive meta-analysis included a total of 4766 dairy products from the 27 studies. The overall molecular prevalence of Listeria monocytogenes in dairy products in Turkey was estimated to be 7.4% (95% confidence interval [CI]: 4.7-10.7%). The I2 value of 93.5% (CI95%91.6–95.0) suggested high heterogeneity, with a τ2 of 3.4422 (CI95%-1.7091–8.5936), and an X2 statistic of 399.2 (P &amp;lt; 0.0001). In conclusion, this meta-analysis reveals a significant seroprevalence of L. monocytogenes in dairy products in Turkey. The findings highlight the need for improved control measures to minimize the risk of contamination and ensure consumer safety. By implementing rigorous hygiene practices, enhancing monitoring systems, and promoting collaborative efforts between industry and regulatory bodies, the dairy sector in Turkey can effectively mitigate the risks associated with L. monocytogenes and maintain high standards of food safety.

Listeria monocytogenes in ready-to-eat (RTE) delicatessen foods: Prevalence, genomic characterization of isolates and growth potential

This study investigated Listeria monocytogenes prevalence and count in 132 ready-to-eat (RTE) delicatessen samples belonging to different categories (starters with/without mayonnaise pasta/rice-based courses, meat/fish-based main courses) produced by an Italian industry. Whole Genome Sequencing characterized the isolates to map the pathogen circulation. Moreover, the growth potential of L. monocytogenes in the most contaminated product was investigated by a challenge test.L. monocytogenes was detected in 23 samples, giving an estimated prevalence of 17.4 %. Starters with mayonnaise showed a very high prevalence (56.7 %), showing the role of the sauce in the diffusion of the pathogen within the plant. A total of 49 isolates were obtained; they belonged to two different serogroups, IIb and IIa, and were related to two clonal complexes (CCs) and sequence types (STs) (CC288-ST330 and CC121-ST717), suggesting the possible persistence and circulation of the pathogen within the plant.The results of the challenge test showed a limited ability to grow in the selected product thanks to the presence of lactic microflora.

Exploring microbial diversity during the artisanal Salchichón production: Food safety in the consumer spotlight

A study was conducted to evaluate the microbiological quality, safety, and variability along the production chain of two artisanal salchichón factories in Andalusia. Aerobic mesophilic bacteria (AMB), total coliforms, coagulase-positive Staphylococcus spp. (CPS), lactic-acid bacteria, moulds and yeasts and the presence of bacterial pathogens were investigated in 3 batches of raw materials, finished products, food-contact surfaces (FCS) and indoor ambient air by using classical microbiological and PCR/PFGE techniques. Of the 171 samples analysed, high inter-batch variability in microbial loads was observed. The prevalence of Listeria monocytogenes and Staphylococcus aureus in samples was 7.01% and 4.67%, respectively. Raw meat and FCS were identified as the potential sources of contamination of the finished products, with 16.67% testing positive for L. monocytogenes and 10% for S. aureus. Diverse sources of L. monocytogenes and a wide range of high pulsotypes were identified in the smaller producer. Although the largest producer showed better hygiene control, L. monocytogenes was detected in salchichón samples with growth-supporting physicochemical values. Therefore, the implementation of food safety management systems, hygiene measures for raw materials and monitoring of hazards are encouraged. Finally, the findings of this study offer valuable and current empirical data that can be used in risk assessments.

Risk factors associated with the prevalence of Listeria monocytogenes in manured soils on certified organic farms in four regions of the United States

IntroductionBiological soil amendments, including raw or untreated manure, are currently used to improve soil fertility, especially in organic operations that prohibit use of synthetic fertilizers. However, addition of untreated manure may pose a risk of contamination of fresh produce by pathogens of public health significance, including Listeria monocytogenes. Organic growers follow United States Department of Agriculture (USDA) National Organic Program regulations for raw manure use, which stipulate that harvest should commence no earlier than 90- or 120-days post-application, depending on direct contact between the edible portion of the produce and the soil. To inform the protection that such time-intervals provide, this study explored the farm-level risk factors associated with L. monocytogenes prevalence in USDA-certified organic farm soils amended with untreated manures.MethodsA longitudinal, multi-regional study was conducted on 19 farms in four states (California, Minnesota, Maine, and Maryland) over two growing seasons (2017 and 2018). Untreated manure, soil, irrigation water, and produce samples were collected and cultured for L. monocytogenes. Mixed effect logistic regression was used to investigate risk factors associated with L. monocytogenes prevalence in soil.Results and DiscussionResults showed that multiple factors influenced the odds of a soil-positive sample, including temporal [year (OR = 0.19), sampling day (OR = 0.09–0.48)] and weather-related [temperature range (OR = 0.48)] variables, manure characteristics [season of application (OR = 0.04, summer), presence of L. monocytogenes (OR = 2.89) and other pathogens in manure (OR = 5.24)], farm management factors [water source (OR = 2.73, mixed), number of year-round staff (OR = 0.02)], and soil characteristics [concentration of generic Escherichia coli (OR = 1.45), moisture (OR = 0.46), organic matter (OR = 7.30), nitrate (OR = 3.07), potassium (OR = 0.09) and calcium (OR = 2.48)]. This study highlights the complexity of L. monocytogenes prevalence in soil and contributes science-based metrics that may be used when determining risk-mitigation strategies for pathogen contamination.

Viability and Virulence of Listeria monocytogenes in Poultry.

The prevalence of Listeria monocytogenes in 30 samples of poultry was determined using culture-dependent (isolation on OCLA and confirmation by conventional polymerase chain reaction -PCR-, OCLA&PCR) and culture-independent (real-time polymerase chain reaction, q-PCR) methods. L. monocytogenes was detected in 15 samples (50.0%) by OCLA&PCR and in 20 (66.7%) by q-PCR. The concentrations (log10 cfu/g) of L. monocytogenes (q-PCR) ranged from 2.40 to 5.22 (total cells) and from <2.15 to 3.93 (viable cells). The two methods, q-PCR using a viability marker (v-PCR) and OCLA&PCR (gold standard), were compared for their capacity to detect viable cells of L. monocytogenes, with the potential to cause human disease. The values for sensitivity, specificity and efficiency of the v-PCR were 100%, 66.7% and 83.3%, respectively. The agreement between the two methods (kappa coefficient) was 0.67. The presence of nine virulence genes (hlyA, actA, inlB, inlA, inlC, inlJ, prfA, plcA and iap) was studied in 45 L. monocytogenes isolates (three from each positive sample) using PCR. All the strains harbored between six and nine virulence genes. Fifteen isolates (33.3% of the total) did not show the potential to form biofilm on a polystyrene surface, as determined by a crystal violet assay. The remaining strains were classified as weak (23 isolates, 51.1% of the total), moderate (one isolate, 2.2%) or strong (six isolates, 13.3%) biofilm producers. The strains were tested for susceptibility to a panel of 15 antibiotics. An average of 5.11 ± 1.30 resistances per isolate was observed. When the values for resistance and for reduced susceptibility were taken jointly, this figure rose to 6.91 ± 1.59. There was a prevalence of resistance or reduced susceptibility of more than 50.0% for oxacillin, cefoxitin, cefotaxime, cefepime ciprofloxacin, enrofloxacin and nitrofurantoin. For the remaining antibiotics tested, the corresponding values ranged from 0.0% for chloramphenicol to 48.9% for rifampicin. The high prevalence and level of L. monocytogenes with numerous virulence factors in poultry underline how crucial it is to follow correct hygiene procedures during the processing of this foodstuff in order to reduce the risk of human listeriosis.

Effect of Sodium Nitrite, Nisin and Lactic Acid on the Prevalence and Antibiotic Resistance Patterns of Listeria monocytogenes Naturally Present in Poultry.

The impact of treating minced chicken meat with sodium nitrite (SN, 100 ppm), nisin (Ni, 10 ppm) and lactic acid (LA, 3000 ppm) on the levels of some microbial groups indicating hygiene quality were investigated. Specifically, aerobic plate counts and culture-based counts of psychrotrophic microorganisms and enterobacteria were obtained. Additionally, the prevalence of Listeria monocytogenes and the resistance of 245 isolates from this bacterium to 15 antibiotics were documented. L. monocytogenes was isolated using the ISO 11290-1:2017 method and confirmed with polymerase chain reaction using the lmo1030 gene. Antibiotic resistance was established using the disc diffusion technique (EUCAST and CLSI criteria). Twenty-four hours after treatment, the microbial load (log10 cfu/g) was reduced (p < 0.05) relative to controls in those samples treated with LA, with counts of 5.51 ± 1.05 (LA-treated samples) vs. 7.53 ± 1.02 (control) for APC, 5.59 ± 1.14 (LA) vs. 7.13 ± 1.07 (control) for psychrotrophic microorganisms and 2.33 ± 0.51 (LA) vs. 4.23 ± 0.88 (control) for enterobacteria. L. monocytogenes was detected in 70% (control samples), 60% (samples receiving SN), 65% (Ni) and 50% (LA) (p > 0.05) of samples. All strains showed resistance to multiple antimicrobials (between 3 and 12). In all, 225 isolates (91.8%) showed a multi-drug resistant (MDR) phenotype, and one isolate (0.4%) showed an extensively drug-resistant (XDR) phenotype. The mean number of resistances per strain was lower (p < 0.01) in the control samples, at 5.77 ± 1.22, than in those receiving treatment, at 6.39 ± 1.51. It is suggested that the use of food additives might increase the prevalence of resistance to antibiotics in L. monocytogenes, although additional studies would be necessary to verify this finding by analyzing a higher number of samples and different foodstuffs and by increasing the number of antimicrobial compounds and concentrations to be tested.

Prevalence of Listeria monocytogenes in milk in Africa: a generalized logistic mixed-effects and meta-regression modelling

Listeria outbreaks and food recalls is on the raise globally. Milk particularly is highly susceptible to Listeria as its production and storage adequately support Listeria growth. The extent of milk contamination with Listeria monocytogenes (Lm) and preventative actions to halt milk associated outbreaks in Africa are unknown. Hence, this study aimed at assessing the national and subregional prevalence of Lm in milk in Africa and identify impacting factors via generalized logistic mixed-effects (GLMEs) and meta-regression modelling. Lm-milk-specific data acquired from primary studies according to standard protocol were fitted using a GLMEs. The GLMEs was subjected to leave-one-study-out-cross-validation (LOSOCV). Factors impacting Lm prevalence in milk were assayed via a 1000-permutation-assisted meta-regression-modelling. The pooled prevalence of Lm in milk in Africa was 4.35% [2.73–6.86] with a prediction interval (PI) of 0.14–59.86% and LOSOCV value of 2.43% [1.62–3.62; PI: 0.32–16.11%]. Western Africa had the highest prevalence [20.13%, 4.13–59.59], then Southern Africa [5.85%, 0.12–75.72], Northern Africa [4.67%, 2.82–7.64], Eastern Africa [1.91%, 0.64–5.55], and there was no record from Central Africa. In term of country, Lm prevalence in milk significantly (p < 0.01) varied from 0.00 to 90.00%. Whereas the Lm prevalence was negligibly different (p = 0.77) by milk type, raw-milk had the highest prevalence [5.26%], followed by fermented-milk [4.76%], boiled-milk [2.90%], pasteurized-milk [1.64%], and powdered-milk [1.58%]. DNA extraction approach did not significantly (p = 0.07) affect Lm prevalence (Boiling [7.82%] versus Kit [7.24%]) as well as Lm detection method (p = 0.10; (ACP [3.64%] vs. CP [8.92%] vs. CS [2.27%] vs. CSP [6.82%]). Though a bivariate/multivariate combination of all tested variables in meta-regression explained 19.68–68.75% (R2) variance in Lm prevalence in milk, N, nation, and subregion singly/robustly accounted for 17.61% (F1;65 = 7.5994; p = 0.005), 63.89% (F14;52 = 4.2028; p = 0.001), and 16.54% (F3;63 = 3.4743; p = 0.026), respectively. In conclusion, it is recommended that adequate sample size should be prioritized in monitoring Lm in milk to prevent spuriously high or low prevalence to ensure robust, plausible, and credible estimate. Also, national efforts/interests and commitments to Lm monitoring should be awaken.

Prevalence of Listeria monocytogenes in RTE Meat Products of Quevedo (Ecuador).

Listeria monocytogenes is a foodborne pathogen that causes listeriosis and can be a problem in areas where meat products are sold at unregulated storage temperatures. In this work, the prevalence of L. monocytogenes was determined in the five most widely traded meat products in the province of Quevedo (Ecuador): bacon, "chorizo paisa", grilled hamburger meat, mortadella, and salami. A total of 1000 samples of these products were analyzed in two seasons of the year (dry season/rainy season). All L. monocytogenes isolates were confirmed by PCR with primers designed for the iap gene. Furthermore, the positive samples were quantified for L. monocytogenes. Of the 1000 meat products analyzed, 163 were positive for L. monocytogenes (16.3%). The prevalence of L. monocytogenes in the two seasons in different meat products was as follows: 22.5% in mortadella, 19% in hamburger meat, 15% in bacon, 14.5% in chorizo paisa and 10.5% in salami. In addition, the concentration of L. monocytogenes in most of the positive samples was in the range of 4-6 log CFU/g or even higher. The results show the need for improvements in the hygienic measures and meat storage temperatures in Quevedo (Ecuador) to avoid risks of foodborne listeriosis.

Prevalence, Serotypes, Antimicrobial Resistance and Biofilm-Forming Ability of Listeria monocytogenes Isolated from Bulk-Tank Bovine Milk in Northern Greece.

The prevalence of Listeria monocytogenes in bovine bulk-tank milk (BTM) in Greece has not been previously investigated. The aim of the study was to estimate the prevalence of L. monocytogenes in bovine BTM in Greece and to characterize the isolates in terms of carriage of genes encoding for pathogenic determinants, assess the isolates' biofilm-forming ability and determine their susceptibility against 12 antimicrobials. Samples (n = 138) of bovine BTM were obtained from farms located throughout Northern Greece and were analyzed qualitatively and quantitatively for L. monocytogenes. Five samples (3.6%) tested positive for L. monocytogenes. The pathogen's populations in these positive samples were below 5 CFU/mL. Most isolates belonged to the molecular serogroup "1/2a, 3a". All isolates carried the virulence genes inlA, inlC, inlJ, iap, plcA and hlyA, but actA was detected in only three isolates. The isolates displayed weak to moderate biofilm-forming ability and distinct antimicrobial resistance profiles. All isolates were characterized as multidrug resistant, with resistance to penicillin and clindamycin being a common feature. Considering that L. monocytogenes constitutes a serious public health threat, the key findings of the study, related to the carriage of virulence genes and multidrug resistance, highlight the importance of continued monitoring of the pathogen in farm animals.

Application of MALDI-TOF Mass Spectrometry for the Assessment of Prevalence of Listeria monocytogenes in Raw Milk, Dairy Products and Freshwater Fishes

Background: Foodborne infections like Listeriosis cover several disorders and are a worldwide public health emergency. Listeria monocytogenes has been isolated from various foodstuffs, including milk and fishes. MALDI-TOF mass spectrometry technique is known for its rapid and accurate identification of bacterial organisms. Methods: In the current research, a total of 360 samples comprising raw milk (130), milk products (125) and freshwater fishes (105) were screened for the detection of pathogenic Listeria species by using the USDA method. The recovered Listeria isolates were characterized using conventional set of biochemical analysis along with sugar fermentation tests and further confirmed by MALDI-TOF MS. The virulent nature of pathogenic Listeria isolates was also assessed by in vitro tests like hemolysis on blood agar, CAMP and PI-PLC assay. Result: In current study, on screening 360 animal origin food samples, three isolates were recovered from raw milk samples and identified as Listeria monocytogenes indicating an overall prevalence of Listeria monocytogenes to the tune of 0.83%. Excellent correlation was observed with identification of Listeria species using conventional phenotypic tests and advanced molecular tool Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) technique. The results depict dependability of advanced technique for rapid and reliable identification of Listeria species.

Prevalence, antibiotic resistance and virulence feature of Listeria monocytogenes isolated from bovine milk in Yunnan, Southwest China

The prevalence of Listeria monocytogenes in bovine milk from different regions of Yunnan Province, Southwest China, was investigated. The isolated strains were genotyped using pulsed-field gel electrophoresis (PFGE), and their virulence and antibiotic resistance potential analysed with whole-genome sequencing (WGS). A total of 8 L. monocytogenes strains were isolated from 4 out of 161 samples, with a detection rate of 2.48%. All strains were at least resistant to one antibiotic, and the majority of strains (75%) were multi-antibiotic resistant. The 8 strains were clustered into 3 pulsotypes, and each pulsotype contained only strains isolated from the same geographical area. One strain was selected from each pulsotype for WGS, and it was found that a total of 99 antibiotic resistance genes and 83 virulence genes were detected from the 3 strains, respectively, which indicated that L. monocytogenes isolated from bovine milk have strong antibiotic resistance and virulence potential.

Salmonella Prevalence Is Strongly Associated with Spatial Factors while Listeria monocytogenes Prevalence Is Strongly Associated with Temporal Factors on Virginia Produce Farms.

The heterogeneity of produce production environments complicates the development of universal strategies for managing preharvest produce safety risks. Understanding pathogen ecology in different produce-growing regions is important for developing targeted mitigation strategies. This study aimed to identify environmental and spatiotemporal factors associated with isolating Salmonella and Listeria from environmental samples collected from 10 Virginia produce farms. Soil (n = 400), drag swab (n = 400), and irrigation water (n = 120) samples were tested for Salmonella and Listeria, and results were confirmed by PCR. Salmonella serovar and Listeria species were identified by the Kauffmann-White-Le Minor scheme and partial sigB sequencing, respectively. Conditional forest analysis and Bayesian mixed models were used to characterize associations between environmental factors and the likelihood of isolating Salmonella, Listeria monocytogenes (LM), and other targets (e.g., Listeria spp. and Salmonella enterica serovar Newport). Surrogate trees were used to visualize hierarchical associations identified by the forest analyses. Salmonella and LM prevalence was 5.3% (49/920) and 2.3% (21/920), respectively. The likelihood of isolating Salmonella was highest in water samples collected from the Eastern Shore of Virginia with a dew point of >9.4°C. The likelihood of isolating LM was highest in water samples collected in winter from sites where <36% of the land use within 122 m was forest wetland cover. Conditional forest results were consistent with the mixed models, which also found that the likelihood of detecting Salmonella and LM differed between sample type, region, and season. These findings identified factors that increased the likelihood of isolating Salmonella- and LM-positive samples in produce production environments and support preharvest mitigation strategies on a regional scale. IMPORTANCE This study sought to examine different growing regions across the state of Virginia and to determine how factors associated with pathogen prevalence may differ between regions. Spatial and temporal data were modeled to identify factors associated with an increased pathogen likelihood in various on-farm sources. The findings of the study show that prevalence of Salmonella and L. monocytogenes is low overall in the produce preharvest environment but does vary by space (e.g., region in Virginia) and time (e.g., season), and the likelihood of pathogen-positive samples is influenced by different spatial and temporal factors. Therefore, the results support regional or scale-dependent food safety standards and guidance documents for controlling hazards to minimize risk. This study also suggests that water source assessments are important tools for developing monitoring programs and mitigation measures, as spatiotemporal factors differ on a regional scale.

Prevalence of Listeria monocytogenes, Salmonella spp., Shiga toxin-producing Escherichia coli, and Campylobacter spp. in raw milk in the United States between 2000 and 2019: A systematic review and meta-analysis

Raw (unpasteurized) milk is available for sale and direct human consumption within some states in the United States (US); it cannot be sold or distributed in interstate commerce. Raw milk may contain pathogenic microorganisms that, when consumed, may cause illness and sometimes may result in death. No comprehensive review for prevalence and levels of the major bacterial pathogens in raw milk in the US exists. The objective of the present research was to systematically review the scientific literature published from 2000 to 2019 to estimate the prevalence and levels of Listeria monocytogenes, Salmonella spp., Shiga toxin-producing Escherichia coli (STEC), and Campylobacter spp. in raw milk in the US. Peer-reviewed studies were retrieved systematically from PubMed®, Embase®, and Web of ScienceTM. The unique complete nonduplicate references were uploaded into the Health Assessment Work Collaborative (HAWC). Based on the selection criteria, twenty studies were included in the systematic review and meta-analysis. Comprehensive Meta-Analysis (CMA) was used for statistical analyses, specifically, random effects meta-analyses were used to synthesize raw bulk tank milk (BTM) and milk filters (MF) data. Data from studies using culture and non–culture-based detection methods were included. Forest plots generated in CMA (Biostat, Englewood, NJ) were used to visualize the results. The average prevalence (event rate) of L. monocytogenes, Salmonella spp., STEC, and Campylobacter spp. in raw BTM in the US was estimated at 4.3% (95% confidence intervals [CIs], 2.8–6.5%), 3.6% (95% CIs, 2.0–6.2%), 4.3% (95% CIs, 2.4–7.4%), and 6.0% (95% CIs, 3.2–10.9%), respectively. Estimated prevalence was generally larger in MF than in BTM. There was not enough data to perform a meta-analysis for the prevalence or levels of pathogens in raw milk from retail establishments or other milk categories.

Listeria monocytogenes isolated from raw milk: phenotypic and molecularcharacterization, pathogenicity testing, and multidrug resistance profiling

Foodborne infections are a worldwide public health emergency that actually impact a variety of disorders. In the current research, a total of 300 samples comprising raw milk (100),milk products (100), and chicken (100) were screened for detection of pathogenic Listeria species by using the USDA method. On conventional biochemical characterization, three isolates were identified as Listeria monocytogenes (from raw milk samples)indicating an overall prevalence of Listeria monocytogenes to the tune of 1%. All other samples comprising milk products and chicken samples (100) showed negativity for the presence of any of the Listeria species. Further, all three isolates were subjected to polymerase chain reaction (PCR) targeting genus-specific (prsA and iap) and species-specific gene (isp) in which all three were turn out positive for both the genes, endorsing their identification as Listeria monocytogenes. All three confirmed Listeria monocytogenes isolates were phenotypically assessed for in-vitro pathogenicity tests like hemolysis on 7% sheep blood agar, CAMP test, and PI-PLC assay. The results revealed their highly pathogenic nature. Subsequently, all these isolates were also assessed for their virulent nature by PCR, targeting the array ofmarkers including virulence-associated genes viz. hlyA, actA, plcA. The results endorsed pathogenic nature of all isolates showing amplification of all targeted virulence genes. The antibiotic resistance profiling revealed occurrence of multidrug-resistant pathogenic L. monocytogenes in foods of animal origin with maximum multiple antibiotic resistance (MAR) index of 0.6and a minimum MAR index of 0.48 in MDR, which is a matter of concern from public health point of view.