Listeria spp. in the raw milk and dairy products in Antakya,Turkey

In this study, the presence of Listeria spp. was investigated in a total of 157 raw milk and dairy product samples sold in Antakya (Antioch). The prevalence of Listeria spp. in raw milk and Turkish white cheese samples was found to be 2.12% and 8.23%, respectively. Listeria monocytogenes was not isolated from raw milk and found in only two cheese samples (2.35%). No Listeria spp. were isolated in any of the butter and yoghurt samples.     

Prevalence and antibiotic susceptibility of Listeria spp. isolated from raw meat and retail foods

Listeria and particularly Listeria monocytogenes is an important foodborne pathogen that can cause listeriosis with flu-like symptoms in healthy people, and severe complications in immunocompromised subjects, children, pregnant women and the elderly. A research survey was conducted to check the presence of Listeria spp. in raw meat and retail products and to analyse their antibiotic resistances. Total prevalence was 11.7%: in raw meat was 21.4%; in ham it was 5.2%; in fresh soft cheese it was 3.49%; in sandwiches it was 5.88%, while we found no isolates in smoked salmon and only two in ready salads (1.23%). The highest percentage of prevalence of L. monocytogenes was found in samples of ham (37.5%), lower percentages were in sandwiches (25.0%), in raw meat samples (23.6%), in fresh soft cheeses (20.0%), while ready salads and smoked salmons were not contaminated. The susceptibility of 168 strains of Listeria spp. was determined by disk diffusion method: we found 51 (30.4%) strains resistant to three or more antibiotics. All isolated strains, except one, are susceptible or at least to one of the first choice antibiotics (ampicillin and gentamycin) or to trimethoprim–sulfamethoxazole used as antibiotic of second choice in the treatment of human listeriosis. Strains isolated from ready-to-eat food show high level of resistance to ampicillin, gentamycin and meticillin. Meticillin is used normally, in treatment of Enterococcus spp. human infection; L. monocytogenes can transfer antibiotic resistance genes from plasmids and transposons to Enterococcus spp. in vitro and in vivo causing an increase of these bacteria resistant to meticillin. L. monocytogenes, in the last decades, is becoming resistant to a lot of antibiotics, a continued surveillance on its incidence on raw foods and on emerging resistances are important to identify food that can represent a risk of infection for the population, particularly for immunocompromised, children, pregnant women and the elderly to ensure effective treatment of human listeriosis with effective antibiotics.     

Incidence of Listeria monocytogenes and Escherichia coli O157:H7 in two Kasar Cheese processing environments

This study was designed to determine the presences of two environmental pathogens in two dairy factories in Sakarya, Turkey. A total of 264 environmental samples, raw milk and cheese samples were taken at four different seasons. According to the results, Listeria monocytogenes or Escherichia coli O157:H7 was isolated from 26 or 2.7% of the samples collected from both factories, respectively. None of the cheese or curd samples were found to be positive for Listeria or E. coli O157:H7. However, 50% of raw milk samples contained Listeria innocua. Listeria was mostly isolated from the swap samples taken from the drains or the floors in processing or packaging areas. However, E. coli was also isolated from the swap samples taken from the workers’ hands and gloves as well as the drains and the floor. Only one raw milk sample contained E. coli O157:H7. A higher prevalence of both pathogens was observed in the summer months than in the other months.     

Incidence of Listeria spp. in fish and environment of fish markets in Northern Greece

The incidence of Listeria spp. in the salt-water edible fish and in the environment of fish markets in Thessaloniki, Northern Greece was studied. One hundred and twenty raw/fresh fish bought at the fish markets of Thessaloniki (Northern Greece), were sampled and tested for presence of Listeria species using a two step enrichment procedure, followed by plating on two selective agars and subsequent biochemical identification of the isolates. Five fish samples were positive for Listeria spp. and in only one sample L. monocytogenes was detected. Also, 100 samples of knives, hands, boxes etc were sampled and 18 samples were positive for Listeria spp. and five for L. monocytogenes. L. innocua was more common being detected in four fish samples and 13 environmental samples. L. seeligeri was detected only in one environmental sample. Our findings indicate that only a few fish were contaminated with Listeria spp., while the level of contamination of the environment of fish markets was higher.     

Prevalence of Listeria spp. and antibiotic susceptibility of Listeria monocytogenes isolated from raw chicken and ready-to-eat chicken products in Jordan

Listeria monocytogenes is the causal agent of listeriosis, a disease that can be serious and is often fatal in susceptible individuals. The objective of the study was to determine the prevalence of Listeria spp. in raw chicken and ready-to-eat (RTE) chicken products in Amman, Jordan and the antimicrobial resistance of L. monocytogenes isolates. A total of 280 raw chicken and RTE chicken products (chicken-shawirma, chicken-burger, chicken-sausage and mortadella) were collected from Amman abattoir and local retail markets in Amman city. Listeria spp. were isolated by the conventional International Organization for Standardization (ISO) method and L. monocytogenes identified by biochemical and Polymerase Chain Reaction (PCR). Results of conventional method showed that out of total 280 samples, 141 (50%) were found to be contaminated with Listeria spp. [L. monocytogenes (18.2%), Listeria ivanovi (26.1%), Listeria grayi (3.5%), Listeria seeligeri (1.8), Listeria welshimeri (0.7%)]. The PCR confirmed all L. monocytogenes isolates (51 isolates: 15 from raw dressed broiler chicken, 23 from chicken-burger, 9 from chicken-sausage, and 4 from chicken-shawirma). Five of the tested L. monocytogenes isolates were resistance to two antibiotics (tilimicosin and tetracycline) among the ten tested antibiotics as determined by microbroth dilution method. The results presented in this study indicate the potential risk of contamination of RTE chicken products with L. monocytogenes.     

Prevalence and antimicrobial resistance of Listeria species isolated from milk and dairy products in Iran

The objective of this study was to determine the prevalence of Listeria spp. in milk and dairy products in Isfahan province, Iran. From March 2007 to September 2009, a total of 594 samples of various milk and dairy products were obtained from randomly selected retail stores. Using conventional bacteriologic method, 55 samples (9.3%) were positive for Listeria spp. The highest prevalence of Listeria was found in raw sheep milk samples (22.6%), followed by cheese samples (18.9%). The most species recovered was Listeria innocua (58.2%); the remaining isolates were Listeria monocytogenes (32.7%) and Listeria seeligari (9.1%). Overall, 54 Listeria isolates (98.2%) were resistant to one or more antimicrobial agents. Resistance to nalidixic acid was the most common finding (96.4%). All Listeria isolates were susceptible to vancomycin. The results of this study indicate the potential risk of infection with Listeria in people consuming raw and unpasteurized milk and dairy products.     

Successful management of Listeria spp. in an avocado processing facility

Listeria monocytogenes can grow and multiply in various food matrices and cause severe human illness. Apart from the influence on consumer health, L. monocytogenes contamination of ready-to-eat (RTE) food products causes major economic losses due to product recalls. Control of foodborne pathogens in RTE food products is a challenge, specifically in foods that cannot undergo a heat-treatment during processing. The aim of this study was to develop control strategies for the management of L. monocytogenes in an avocado processing facility, additional to a quality control system. An in-house monitoring system (IMS) was established to test specifically for Listeria spp. in the final products and processing environment, including floors, equipment, work areas and personnel. Guacamole and environmental samples were collected and tested on-site for Listeria with the ISO 11290-1 method. Based on the prevalence of Listeria, the facility introduced new strategies in processing to counter cross contamination. Results from the 2014 guacamole production season showed almost complete eradication of Listeria spp. in final products (0.17%, n = 1170) and the processing facility (0.79%, n = 1520). This is a major achievement since the highest incidence of Listeria spp. over a period of five years was measured at 11.39% (n = 948) in the final product during the 2013 season and 13.44% (n = 1927) in the processing facility in 2011. These results indicate that successful management of Listeria spp. in an avocado processing facility can be accomplished with in-house monitoring of the listerial population and subsequent adjustments to the processing system.     

Occurrence and traceability of Listeria monocytogenes strains isolated from sheep's milk cheese-making plants environment

The aim of the study was to conduct an extensive survey on Listeria monocytogenes and Listeria spp. environmental contamination in 13 cheese-making plants. A total of 409 environmental and food samples were collected during years 2011–2013. Listeria spp. contamination was observed in all the facilities, while L. monocytogenes was recovered from 12 facilities with a prevalence ranging between 3.0% and 22.6%. Floor drains were the most contaminated sampling sites (48.8% of positive samples), serving as harbourage site for subsequent contamination. Out of 616 isolates, 277 (45.0%) were Listeria innocua, 274 (44.5%) L. monocytogenes, 41 (6.6%) Listeria ivanovii, 14 (2.3%) Listeria welshimeri and 10 (1.6%) Listeria gravyi. Serotyping carried out by PCR and agglutination method for L. monocytogenes revealed that 169 strains (61.7%) were serotype 1/2a, 65 (23.7%) 4b, 20 (7.3%) 1/2b, 10 (3.6%) 3a, 7 (2.5%) 1/2c and 3 (1.1%) 3b. PFGE conducted on L. monocytogenes isolates using AscI and ApaI restriction enzymes, yielded 6 clusters. Two predominant PFGE clusters were observed including respectively 36 and 32 strains. Within cheese-making plants, L. monocytogenes showed wide variability with strains distributed up to 4 different clusters. Pulsotypes isolated from raw milk filter were never detected in the processing environment, indicating that the contamination originated from sources other than raw milk. The isolation of strains with similar profile from different sampling sites, within and among cheese-making plants, indicated the possible transfer of L. monocytogenes contamination along production lines and from one facility to another. Strains recovered from food were confirmed as originating from the processing environment.     

Prevalence of Listeria spp. at a poultry processing plant in Brazil and a phage test for rapid confirmation of suspect colonies

Sites and occurrence of Listeria contamination in an industrial poultry processing plant were investigated by sampling carcasses at varying stages of processing and testing the hands and gloves of food handlers as well as the chilling water used in the process. In the course of nine visits to a local processing plant we collected a total of 121 samples: 66 from carcasses, 37 from workers' hands and gloves and 18 from the water used for chilling. Except for the water samples Listeria was isolated at all sampling sites. The species most often isolated was Listeria innocua, which accounted for 28 of the 31 (90.3%) isolates. The frequency of Listeria in the chicken carcasses was similar at bleeding, defeathering and end of evisceration stages (33.3%), reduced during scalding (16.7%), and rose immediately after initial evisceration stage (50%) to peak after packaging (76.2%). The carcasses were contaminated by L. monocytogenes serotypes 1b and 1c only during packaging. The prevalence of Listeria spp. on workers' hands and gloves was 46% mostly with L. innocua (40.5%) followed by L. monocytogenes 1b (11.8%). Chilling water presented more than 100 ppm of chlorine, which could explain why the samples were negative to Listeria. As the contamination by Listeria in the carcasses progressively rose both in number, species and strains during processing it seems reasonable to conclude that those carcasses become contaminated at the processing level. Improvement and innovation measures to control bacteria in general at the processing plant level are necessary to effectively reduce final product contamination by L. monocytogenes. In the course of this work we introduced a bacteriophage susceptibility test to confirm suspected Listeria colonies which was able to reduce the time of analysis to a minimum of 30 h depending on the isolation technique employed.     

Prevalence of Listeria spp. in ready to eat foods (RTE) from Algiers (Algeria)

The aim of this study was to establish the occurrence of Listeria spp., especially Listeria monocytogenes in ready to eat RTE food marketed in Algiers (Algeria).A total of 227 samples were collected from different producers and retailers.All samples were analyzed using a conventional cultivation method AFNOR V08-055.Out of 227 samples tested, 21 (9.3%) tested positive for Listeria spp. among them, 6 (2.6%) tested positive for L. monocytogenes. L. innocua was the most common Listeria species found being detected in 11 samples (4.8%), although both Listeria ivanovii and Listeria welshimeri were detected in 3 (1.3%) and 1 (0.4%) food samples respectively.The study of the antimicrobial sensitivity of Listeria monocytogenes strains showed no resistance.The study has enabled us to detect these contaminants in a wide range of RTE foods, to suggest that contamination likely occurs after heat treatment, and to assess the danger represented by this category of food for populations at risk.     

Comparison of two pre-enrichments broths for recovering Listeria spp. from salmon (Salmo salar) and salmon-trout (Oncorhynchus mykiss)

Low levels of occurrence of Listeria spp. in fresh salmon (Salmo_salar ) and salmon-trout (Oncorhynchus mykiss), may be related to the selectivity of the pre-enrichment broth recommended by ISO 11290-1. The purpose of this study was to compare the abilities of Fraser base (without supplements) and 0.1% (w/v) peptone water for recovering Listeria spp. from the fresh fish samples.Fifty-six fish were swabbed and the swabs placed in Fraser base and in 0.1% (w/v) peptone water. Samples were analysed 4–6 h later following the ISO 11290-1 protocol. A total of fifteen Listeria spp. positive samples were found. Three and twelve samples were found to contain respectively, L. monocytogenes and L. innocua. The Fraser base did not detect any of the three L. monocytogenes positive samples. Only two Listeria spp. positive samples were simultaneously recovered by the two broths.     

Incidence and genetic variability of Listeria species from three milk processing plants

The presence of Listeria in three milk processing environments as a potential source of milk contamination was assessed. Swab samples (n = 210) taken from milk processing plants were examined. Sample sites included the milk processing equipment, besides areas handling raw and pasteurized milk. The USDA Listeria-selective enrichment procedure was used to process the samples. Forty one (19.52%) Listeria isolates were recovered. The isolates were further subjected to biochemical and genotypic characterization. Out of 41 isolates, 16 (7.62%) were confirmed as Listeria monocytogenes, 2 (0.95%) as L. ivanovii, 19 (9.05%) as L. innocua. 1 (0.48%) as L. seeligeri and 3 (1.43%) as L. grayi. All the L. monocytogenes isolates were positive for the hlyA gene. PCR based serotyping revealed all L. monocytogenes to be of 1/2a, 1/2c, 3a and 3c serovar group. AscI and ApaI restriction analysis yielded four PFGE clusters for 16 L. monocytogenes isolates obtained from raw milk collector, milk silos, buttermilk mixer, cheese and other milk product processor. No predominant PFGE cluster was observed among these L. monocytogenes isolates. The main sources of L. monocytogenes were found to be raw milk collector and milk silos. In the present study L. monocytogenes was isolated from milk and milk products processing plants which could cross-contaminate the processed products and may possess a potential threat to public health.     

Investigation on prevalence of Listeria spp. and Listeria monocytogenes in animal-derived foods by multiplex PCR assay targeting novel genes

Chilled and frozen animal-derived food can be contaminated by Listeria spp., emerging foodborne pathogens in food industry. The objective of this study was to mine novel target genes by comparative genomics approach for multiplex PCR detection and differentiation of Listeria monocytogenes and other Listeria spp. in food. Multiplex PCR assay targeting the genetic markers LMOf2365_2721, AX25_00730, lin1814, int, lwe1673, and Oxidoreductase gene, resulted in the amplification of DNA fragments of 583 bp, 703 bp, 421 bp, 994 bp, 345 bp, and 201 bp from L. monocytogenes, Listeria ivanovii, Listeria innocua, Listeria seeligeri, Listeria welshimeri, and Listeria grayi, respectively. The detection limits of the multiplex assays were as low as 89 fg/μL genomic DNA and 910 CFU/mL of bacterial culture. The prevalence of Listeria spp. was determined using the developed multiplex PCR assay and standard microbiological method in a total of 200 food samples collected from different supermarkets and traditional agri-product markets in Nanjing, China. A total of 28 samples were found to be positive for the presence of Listeria, including 10.9% (6/55) of livestock meat samples, 22% (11/50) of poultry samples, 15% (6/40) of shellfish samples, 13.3% (4/30) of octopus samples and 4% (1/25) of freshwater fish samples. Of these, 13 isolates were classified as L. monocytogenes, 11 were classified as L. innocua, 2 were classified as L. ivanovii and 3 were classified as L. welshimeri. These results demonstrate that the multiplex PCR assay based on novel target genes is able to rapidly detect the Listeria spp. in 12 h with high accuracy and sensitivity, which may be used in the future for detection of Listeria spp. in animal-derived food products.     

Prevalence and antimicrobial resistance patterns of Listeria species isolated from poultry products marketed in Iran

In the present study, a total of 402 poultry product samples composed of raw, ready-to-cook (RTC) and ready-to-eat (RTE) products were examined for the presence of Listeria spp. The total contamination rate with Listeria spp. in poultry products was 33.3% with a higher rate of contamination in warm seasons than in cold seasons. The most species recovered was Listeria innocua (46.3%); the remaining isolates were Listeria monocytogenes (38.8%), Listeria ivanovii (9.7%) and Listeria seeligeri (5.22%). L. monocytogenes was detected in 14.1%, 12.2% and 11.4% of raw, RTC and RTE poultry products, respectively. Serotype 4b (44.9%) was the predominant serotype of L. monocytogenes isolates followed by 1/2a (40.8%), 1/2b (10.2%) and 1/2c (4.08%). Considering seasonal variability, 1/2a was the most prevalent serotype in warm seasons, while 4b was predominant in cold seasons. The Listeria spp. particularly L. monocytogenes isolates were highly resistant to ampicillin, penicillin, fluroquinolones and tetracycline. The results indicate that high prevalence of Listeria spp. especially L. monocytogenes in poultry products, and resistance of the isolates to the antimicrobials commonly used to treat human listeriosis could be a potential health hazard for consumers. In addition, prevalence of L. monocytogenes serotype 4b that involved in the majority of foodborne outbreaks of human listeriosis is a public health concern.     

Inhibitory effects of chitosan in combination with antibiotics on Listeria monocytogenes biofilm

Microorganisms on living or inert surfaces usually form biofilms which makes microbes highly resistant to antibiotics and immune clearance. Herein we investigated the synergistic effect of several antibiotics with chitosan on eradication biofilms built by Listeria monocytogenes, a causative organism of the serious foodborne illness in a wide variety of mammalian species including human and food animals. Our data demonstrated that chitosan combined with the aminoglycoside antibiotic such as amikacin, but not clindamycin, vancomycin and erythromycin was more effective in inhibition or disruption of L. monocytogenes biofilms than the antibiotic alone did. To elicit an optimal anti-biofilm and bactericidal efficiency, chitosan with specialized molecular size (∼13 kDa) and high N-deacetylation degree (∼88%) was required in this combination. Further evidence showed that this strategy was effective in removing biofilms built by multiple L. monocytogenes strains and other Listeria species. Thus, our data highlighted that chitosan/amikacin combination might be useful for the treatment of Listeria biofilms and help prevent the spread of antibiotic resistance through improving antibiotic effectiveness.     

Influence of Listeria innocua on the growth of Listeria monocytogenes

The growth of Listeria monocytogenes and Listeria innocua strains was monitored during this study: (i) in TSB–YE media and (ii) in a food matrix (pasteurized milk) according to the ISO 11290-1 methodology. Different inocula concentrations and mixtures were tested. The response was shown to be strain dependent. In TSB–YE the inhibition of a L. monocytogenes strain was observed in just one of the three mixtures (L. monocytogenes_1340 with L. innocua_11288) showing a reduction of 1.37 log cfu/ml after 42.5 h and 1.85 log after 66.5 h of incubation. In pasteurized milk the inhibition of L. monocytogenes by L. innocua was always observed when L. innocua was present in higher concentrations than L. monocytogenes. The reverse was also observed but only in one mixture (cocktail of six L. monocytogenes with L. innocua_2030c) when the initial concentration of L. monocytogenes was 100 times higher than L. innocua suggesting the phenomenon of quorum sensing. Furthermore, inhibitory activity was not caused by bacteriocins, and no correlation between the growth rate and inhibition was demonstrated.     

Listeria monocytogenes in ready-to-eat vacuum and modified atmosphere packaged meat and fish products of Estonian origin at retail level

The prevalence, counts and genetic diversity of Listeria monocytogenes in ready-to-eat (RTE) vacuum and modified atmosphere packaged meat and fish products was studied in Estonia. Within two consecutive years 370 RTE food samples were collected at retail level from which 11% were found to be positive for L. monocytogenes. Contamination was higher among RTE fish products (17%) than in RTE meat products (6%). Generally, the counts of L. monocytogenes in positive products remained under ten colony forming units (CFU) per gram of product. Only 1.6% of the RTE meat and fish products contained L. monocytogenes in range of 10–100 CFU/g and 0.3% more than 100 CFU/g at the end of shelf-life. The food category containing highest L. monocytogenes prevalence was RTE lightly salted fish products with the prevalence of 32%. Only one (0.3%) RTE food sample exceeded the 100 CFU/g food safety criterion set out in the EU Regulation 2073/2005. Pulsed-field gel electrophoresis (PFGE) characterization of the isolates showed an overall similarity higher than 70%, and nine clusters based on 100% similarity were revealed. PFGE genotyping revealed that the few predominant pulsotypes were associated with particular food plants.     

Changing regulation: Canada's new thinking on Listeria

In 2008, Canada experienced one of its most serious outbreaks of foodborne listeriosis. During this outbreak, which was eventually traced back to contaminated deli-meat, there were 57 cases of illness and 23 deaths. As a result of the outbreak, the Canadian Food Inspection Agency (CFIA) has developed new Directives for the registered meat and poultry sector. Under the new requirements, production facilities must implement food contact surface testing for Listeria spp. and/or Listeria monocytogenes. The enhanced requirements focus on early detection and control of L. monocytogenes by introducing new testing and reporting requirements for industry, e.g., positive test results from all food contact surface testing must now be immediately reported to the CFIA and companies must perform trend analysis on their test results.In addition, work has begun on updating the current Health Canada policy on L. monocytogenes. The Canadian approach to Listeria control stresses the importance of environmental sampling. In addition to government agencies and food industries working diligently at minimizing the exposure to L. monocytogenes, consumers also have an important role to play in the farm-to-fork continuum. That role calls for Canadians to learn and adopt safe food handling, avoidance of certain high-risk foods, and preparation practices. To this effect, Health Canada has and will continue to undertake the development of science-based consumer education material which will help create an understanding of food safety issues within the context of the public’s right to know about the potential dangers in food, and industry’s responsibility for producing safe food. A combination of all these approaches are currently being adopted and/or developed to improve the control of L. monocytogenes in RTE foods sold in Canada.     

Polymerase chain reaction detection of Listeria monocytogenes using oligonucleotide primers targeting actA gene

A polymerase chain reaction (PCR) assay targeting the gene encoding actA was developed for detecting Listeria monocytogenes in pure cell cultures and on artificially contaminated pork, water and milk. 827-bp PCR product was detected in all 51 L. monocytogenes strains belonging to four different sero-groups (1/2a, 1/2b, 1/2c, and 4b). In contrast, the PCR product was not detected both in all other Listeria spp., including Listeria innocua, Listeria ivanovill, Listeria seelingeri, Listeria welshimeri, or Listeria grayi and in non-Listeria bacteria, indicating that the primer set we use was highly specific for L. monocytogenes. The detection limit of the PCR assay for pure cell cultures was 10⁵ cfu per ml pure cell culture. However, the assay could detect as few as 10¹ cfu of L. monocytogenes in 25 g of pork and 25 ml milk and water following 16 h of enrichment in Listeria Enrichment broth (LEB) at 30 °C. Only a large number of dead L. monocytogenes cells can cause false positivity, as determined using model pork, milk and water samples artificially contaminated with decimal dilutions of dead L. monocytogenes. The total assay time including enrichment was approximately 18 h. These results suggest that the PCR assay based on amplifying actA gene can used to rapidly detect L. monocytogenes on pork, milk, water and possibly other types of food products.     

Prevalence of L. monocytogenes and Salmonella spp. in Tulum cheese

Tulum cheese, which is produced from raw milk, is one of the most popular semi-hard cheeses in Turkey. The growth of some food pathogens such as Listeria monocytogenes and Salmonella spp. in cheese and other dairy products may cause serious health problems for consumers. The aim of this study was to assess the presence of L. monocytogenes and Salmonella spp. in Tulum cheese sold in Istanbul. During the period March 2004–March 2005, a total of 250 Tulum cheese samples were obtained from various markets located in Istanbul and the presence of L. monocytogenes and Salmonella spp. was analyzed according to “The US Food and Drug Administration” (FDA) methods. The results were positive for L. monocytogenes and Salmonella spp. in 12 (4.8%) and 6 (2.4%) samples respectively.