Food Safety Awareness, Changes in Food Purchasing Behaviour and Attitudes towards Food Waste during COVID-19 in Türkiye.

(1) Background: The COVID-19 pandemic brought the key issues of food security, food safety, and food waste into sharp focus. Türkiye is in the enviable position of being among the top ten agricultural economies worldwide, with a wide diversity of food production. This survey was undertaken in order to gain insights into consumer behaviour and attitudes in Türkiye with respect to these issues. The objective was to highlight strengths and weaknesses, identify areas for improvement, and present strategies for the future. (2) Methods: This survey was carried out between April and May 2022 in 12 provinces throughout Türkiye. Face-to-face interviews were performed with 2400 participants representing a cross-section of ages, educational attainment, and socio-economic categories. The findings were evaluated statistically. (3) Results: The results provide an insight into attitudes and behaviours, both pre-COVID-19 and during the pandemic. In several ways, the pandemic enhanced knowledge and improved behaviour, leading to improvements in diet and reductions in food waste. However, worrying concerns about food safety persist. Specific attention has been given to understanding patterns of bread consumption, particularly in consideration of waste. (4) Conclusions: It is hoped that the results of this survey will increase dialogue between the components of the food sector, encourage education initiatives, and contribute to improving food safety and security and reducing food waste in Türkiye and beyond.

Molecular characterization of extended-spectrum ß-lactamases-producing Enterobacteriaceae species in ground beef and chicken meat.

The objectives of this study were i) to characterize extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-E) using pheno- and genotyping methods, ii) to evaluate the antimicrobial resistance pattern against 10 antibiotics, and iii) to investigate class 1 integron (intI1) in 80 Enterobacteriaceae isolates obtained from chicken meat (n = 40; 47 isolates) and ground beef (n = 40; 33 isolates) samples. Through the study, we found that 55 (68.7 %) of 80 Enterobacteriaceae isolates were capable of ß-lactamase activity, and 38 (47.5 %) of them were multi-drug-resistant (MDR). The ground meat-origin isolates are 1.2 times more likely to produce imipenem resistance compared to chicken-meat-origin isolates (z = 2.1, p < 0.05, OR = 1.42). ESBL-E was found in 18 (22.5 %) of the isolates, 16.3 % of chicken meat and 6.3 % of ground beef origin. The bla genes were detected in 14 isolates [bla-TEM (n = 10; 12.5 %); bla-SHV (n = 4; 5.0 %); bla-CTX-M (n = 0)], where the predominant species were Escherichia (E.) coli and Citrobacter braakii. The nine ESBL-E isolates were MDR. Twenty-eight (35.0 %) of 80 isolates were found to be resistant to at least one third-generation cephalosporin, and eight (28.6 %) of them were also ESBL-E. Eleven of 16 (48.5 %) carbapenem-resistant isolates were ESBL-E. The intI1 gene was found in 13 (16.3 %) isolates, five of which were ESBL-E, and four of which were MDR. Co-existing with bla-TEM and the intI1 isolate was ESBL-E. coli, which was resistant to nine antibiotics. In conclusion, chicken meat and ground beef may pose a potential risk of containing ESBL-E, and bla genes which could be spread to the entire food chain.

Presence of quorum sensing system, virulence genes, biofilm formation and relationship among them and class 1 integron in carbapenem-resistant clinical Pseudomonas aeruginosa isolates.

Carbapenems are the most effective agents for treating clinical P. aeruginosa (PsA) infections. During an infection, a quorum-sensing (QS) system and its regulating virulence genes have a great role. The aim of the study was to detect the presence of a las and rhl QS system and related virulence genes, biofilm formation and a class 1 (Cls1) integron. A total of 52 carbapenem-resistant PsA (CRPsA) isolates obtained from Kastamonu, Turkey was analyzed. For the isolation and identification of CRPsA isolates, a conventional culture method, an automated VITEK-2 compact system, and oprL gene-based molecular technique were applied. The two QS system genes were detected in 51 (98.1%), and co-existed of four two QS system genes (lasI/R and rhIl/R genes) were determined in 41 (78.8%) of the isolates. algD, lasB, toxA and aprA genes were detected in between 46.1 and 88.5%, and co-existence of four two QS system genes with four virulence genes were detected in 40.4% of the isolates. Biofilm formation using microtiter plate assay and slime production using Congo Red Agar and Cls1 integron were determined in 84.6%, 67.3% and 51.9% of the isolates, respectively. According to statistical analyses results, there was a significant positive correlation (p < .10) between the las and the rhl systems and a strongly and positive correlation (p < .01 or p < .05) between the rhl system-three virulence genes and slime production-and among some virulence genes. In conclusion, the CRPsA isolates tested in the study are highly virulent and QS systems have a significant role in pathogenesis.

Quorum sensing systems and related virulence factors in Pseudomonas aeruginosa isolated from chicken meat and ground beef.

The objective of this study was to evaluate 50 [chicken meat (n = 45) and ground beef (n = 5)] Pseudomonas aeruginosa isolates to determine the expression of the lasI and rhl QS systems, related virulence factors, and the presence of N-3-oxo-dodecanoyl homoserine lactone (AHL: 3-O-C12-HSL). For the isolation and identification of P. aeruginosa, conventional culture and oprL gene-based molecular techniques were used. In relation to QS systems, eight genes consisting of four intact and four internal (lasI/R, rhlI/R) genes were analyzed with PCR assay. The two QS systems genes in P. aeruginosa isolates from ground beef (80.00%) and chicken meat (76.00%) were present at quite high levels. The 3-O-C12-HSL was detected in 14.00% of the isolates. Both biofilm formation and motility were detected in 98.00% of the isolates. Protease activity was determined in 54.00% of the isolates. Pyocyanin production was detected in 48.00% of the isolates. The las system scores strongly and positively correlated with the rhl system (p ˂ .01). PYA moderately and positively correlated with protease (p ˂ .05). In addition, there was statistically significance between lasI and protease activity (p < .10), and rhlI and twitching motility (p < .10). In conclusion, the high number of isolates having QS systems and related virulence factors are critical for public health. Pyocyanin, protease, and biofilm formation can cause spoilage and play essential role in food spoilage and food safety.

Quorum sensing systems, related virulence factors, and biofilm formation in Pseudomonas aeruginosa isolated from fish.

In this study, two quorum sensing (QS) system genes, las and rhI; N-3-oxo-dodecanoyl homoserine lactone (AHL; 3-O-C12-HSL); and QS-related virulence factors and correlation between them were assessed in 30 fish origin P. aeruginosa isolates. The detection of two QS system of the isolates, and eight gene regions consisting of four intact (lasI/R, rhlI/R) and four internal (lasI/R, rhlI/R) genes were tested by PCR assay. According to findings, las and rhI QS system genes were detected in 27 and 30 isolates, respectively, while 3-O-C12-HSL was determined in 13 isolates. A total of 22, 27, and 18 isolates were capable of pyocyanin production, protease, and elastase activity, respectively. Biofilm formation was detected using three methods in all 30 isolates: 12 by Congo red agar, 14 by microtiter plate, and 29 by tube test. Twitching and swarming motility types were detected in 30, but the swimming motility was determined in 25 isolates. The rhI QS system genes detected in all of the isolates having three types including motility, PYA production, and protease and elastase activities. The las QS system genes were detected in 27 of the motility, 17 of PYA production, 25 of protease, and 16 of elastase activity having isolates. In conclusion, the high number of P. aeruginosa isolates from fish tested have two QS systems and related virulence factors. There was also correlation between them.

Prevalence and Antibiotic Resistance of Salmonella Enteritidis and Salmonella Typhimurium in Ground Beef and Meatball Samples in Samsun, Turkey.

The aims of this study were to evaluate the prevalence of Salmonella spp., including S. Enteritidis and S. Typhimurium, their antibiotic resistance profiles, and the presence/absence of class 1 integron (intI1) in 50 raw ground beef and 50 raw, meatball samples collected in the Samsun Province, Turkey. For the detection of Salmonella, conventional culture technique and PCR assay were used. The antibiotic resistance profiles of the isolates against nine antibiotics were tested. Salmonella spp. was detected in 20 (n = 86 isolates) samples, namely 12 ground beef and 8 meatball samples. Salmonella Enteritidis (n = 12; 24 isolates) or S. Typhimurium (n = 3; 6 isolates) was detected in 15 (75.00%, n = 30 isolates) samples. At least one species-specific gene (oriC or invA) was detected in the isolates. All isolates were sensitive to two of the third-generation cephalosporins and also nalidixic acid. There was a different level of multidrug resistance (MDR) between S. Enteritidis and Typhimurium isolates. Class 1 integron was detected in four samples (n = 7 isolates); seven isolates were S. Enteritidis and four out of the seven S. Enteritidis isolates were also MDR. In conclusion, the presence of Salmonella, particularly S. Enteritidis and S. Typhimurium, in ground beef and meatballs may cause foodborne infections. The presence of antibiotic-resistant Salmonella and S. Enteritidis with the Cls1integron is important for horizontal antibiotic gene transfer.

Plasmid-Mediated Colistin Resistance in Escherichia coli O157:H7 Cattle and Sheep Isolates and Whole-Genome Sequence of a Colistin-Resistant Sorbitol Fermentative Escherichia coli O157:H7.

The aims of this study were to investigate the plasmid-mediated colistin resistance genes (mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5), phenotypic colistin resistance in Escherichia coli O157:H7+/H7- strains isolated from cattle and sheep, and whole-genome sequence (WGS) analysis of colistin-resistant sorbitol fermentative E. coli O157:H7. According to the results, 5 of the 49 isolates were found to harbor mcr-2 and/or mcr-3 genes. Three isolates, including a sorbitol fermentative E. coli O157:H7, were found phenotypically resistant to colistin with a minimum inhibitory concentration value of 128 µg/mL. The genome of sorbitol fermentative E. coli O157:H7 did not show 100% similarity to any of the other genome sequences found in the universal genome database. It has also been determined that this isolate carried 62 different antimicrobial resistance genes. This is the first report of plasmid-mediated mcr-2 and mcr-3 genes carrying E. coli O157:H7 from cattle and sheep isolates and WGS of a colistin-resistant sorbitol fermentative E. coli O157:H7. Findings of this study indicate that cattle and sheep can be an important source of colistin resistance in E. coli O157:H7, and slaughterhouse wastewater might be a significant route for dissemination of the plasmid-mediated colistin genes. Therefore, the use of colistin in veterinary medicine should be restricted to reduce the development of resistance. Also it may be necessary to review the non-sorbitol fermentation-based isolation protocol for not missing the sorbitol fermentative E. coli O157:H7 in epidemiological studies.

Pathogenicity, genotyping and antibacterial susceptibility of the Listeria spp. recovered from stray dogs.

The present study aimed to determine the prevalence of Listeria spp. in stray dogs in the Kayseri province of Turkey. In addition, serotyping, genotyping and virulence gene analysis of the isolated Listeria spp. were performed and their pathogenicity and antibacterial susceptibility were investigated. The study included 80 rectal swaps taken from 80 stray dogs of different ages and gender that were sheltered in the Kayseri Municipal Dog Shelter. Listeria selective broth and Listeria selective agar were used for the isolation of Listeria spp. and the isolates were identified using a Microbact 12L (Oxoid, England) identification test kit. 16S rDNA sequencing and species-specific polymerase chain reaction (PCR) were performed for molecular identification of the isolates, multiplex PCR and a serological test were performed for serotyping, and PCR was used for virulence gene analysis. For determining the pathogenicity of L. monocytogenes and L. innocua isolates, a total of 100 mice (50 pregnant and 50 non-pregnant) were used. The mice were infected intraperitoneally; the inoculation dose was 1 × 109 CFU/mL and 0.2 mL was used for each animal. Tissue samples obtained from infected mice were processed for the re-isolation of the Listeria spp. and then stained with hematoxylin eosin and Brown-Brenn Gram stain. The antibiotic susceptibilities of the isolates were determined by the disk diffusion method. Listeria spp. were isolated from 5 (6.25%) of the 80 fecal samples. While 1 of the isolates was identified as L. monocytogenes, 4 of them were identified as L. innocua. Serotyping by serological and molecular methods revealed the isolate of L. monocytogenes to be serotype 1/2a. According to the phylogenetic trees, L. innocua and L. monocytogenes strains were clustered in different groups. The L. monocytogenes isolate was positive for all virulence genes tested. All L. innocua isolates were positive for the plcB gene. While all L. innocua isolates were negative for the lin1068 gene, 3 L. innocua isolates were found to be positive for the lin0558 gene. In mice infected with L. monocytogenes, pathological findings were observed in the uterus, intestines, pancreas, and heart. In mice infected with L. innocua, pathological findings were observed in the stomach, intestines and spleen. L. monocytogenes- or L. innocua-related infections or other inflammatory reactions were not observed in the brains of infected animals. On histopathological examination with Gram stain, an abundance of Listeria spp. was observed in the lesions of the liver, spleen, uterus, and kidney. Moreover, while abortion was observed in all animals infected with L. monocytogenes, it was not observed in any of the animals infected with L. innocua. Antibiotic susceptibility testing revealed that all 5 isolates were sensitive to ampicillin, amoxicillin/clavulanic acid, erythromycin, gentamicin, penicillin G, and trimethoprim-sulfamethoxazole and were resistant to nalidixic acid, streptomycin, and cefuroxime sodium. Considering also the pathogenicity of the isolated microorganisms, it can be suggested that stray dogs as carriers of Listeria spp. are a significant risk to public health. As L. innocua isolates, which are considered apathogenic, led to the occurrence of lesions similar to those caused by L. monocytogenes, detailed studies on the pathogenesis of L. innocua infections caused by L. innocua isolates recovered from various sources are required.

Prevalence and Characterization of Listeria monocytogenes Isolated from Beef and Sheep Carcasses in Turkey with Characterization of Locally Isolated Listeriophages as a Control Measure.

Swab samples from cattle and sheep carcasses (120 of each) were tested for Listeria monocytogenes, and 120 slaughterhouse wastewater samples were tested for listeriophages over 12 months (10 samples per month) to note the seasonal distribution. L. monocytogenes and bacteriophage isolates were characterized, and the biocontrol of L. monocytogenes was investigated in meatballs with a phage cocktail. L. monocytogenes was found in 3.4 and 2.5% of cattle and sheep carcasses, respectively. All the isolates were found to harbor hlyA, actA, inlA, inlB, inlC, inlJ, plcA, plcB, fbpA, and fri genes with varied mRNA expression levels by real-time reverse transcriptase PCR analysis. Five isolates did not harbor the vip gene. According to enterobacterial repetitive intergenic consensus PCR, L. monocytogenes isolates were classified into four different groups based on their DNA patterns. The L. monocytogenes isolates were characterized for antibiotic susceptibility; one strain was found to be resistant to five different antibiotic classes. Of 11 lytic listeriophages, two were selected for the cocktail based on their DNA restriction profiles, efficiency of plating, transmission electron microscopy, and in vitro and in vivo analyses. In the biocontrol study, we used a food model that consisted of a novel bacteriophage cocktail in raw meatballs. The highest reduction of L. monocytogenes was recorded as 2.2 log CFU/g at a multiplicity of cellular infection of 4.7 at the end of 1 h. In conclusion, the new bacteriophage cocktail in this study can be considered an efficient biocontrol agent of L. monocytogenes in meatballs.

Prevalence and Molecular Characterization of Methicillin-Resistant Staphylococcus aureus in Foods of Animal Origin, Turkey.

In the present study, 175 coagulase-positive Staphylococcus (CPS) isolates recovered from samples of beef (n = 110), raw milk n = 56), and fish (n = 9) were analyzed for methicillin resistance using MIC and PCR assays. Methicillin-resistant (MR) Staphylococcus aureus (SA) isolates were then characterized using pulsed-field gel electrophoresis (PFGE). According to findings, 62 (35.4%) of the isolates (44 from beef, 9 from milk, and 9 from fish) were identified as S. aureus based on the presence of the nuc gene. MRCPS was detected in 18 (10.3%) of 175 CPS isolates based on the presence of the mecA gene. Among these isolates, 15 (24.2%) were MRSA: 4 (26.7%) from beef, 2 (13.3%) from milk, and 9 (60%) from fish. However, based on the MIC assay, 21 (12.0%) of the CPS isolates (1 from beef, 15 from milk, and 5 from fish) were MRCPS, indicating a discrepancy between the results of these two methods. The PFGE results indicated genetic heterogeneity of the isolates; six PFGE clusters were found. These results confirm that MRSA is present in foods of animal origin, which is a concern to human health, and indicate the importance of method selection for determination of methicillin resistance. The identity of MR isolates should be verified by PCR to obtain more reliable results.

Prevalence and characterization of Salmonella isolated from chicken meat in Turkey.

This study was conducted in a Turkish province to investigate the presence of Salmonella spp. in 150 chicken meat samples using 2 phenotyping techniques: classic culture technique (CCT) and immunomagnetic separation (IMS). For the confirmation of the isolates at molecular levels, invA gene was detected in these isolates. The presence of invA, class 1 (Cls1) integrons, and integrase (Int1) genes was demonstrated by PCR assay; and the resistance of the isolated Salmonella spp. strains to antibiotics was determined by disk diffusion test. All the cultural and PCR results were evaluated together; Salmonella spp. were detected in a total of 64 (42.66%) chicken meat samples. Contamination rate was higher in carcasses (53.33%, n = 75) than in meat pieces (32%, n = 75). When results of standard culture were compared with IMS technique, IMS (n = 54) showed a clear superiority over the CCT (n = 38). A very high resistance rate (≥ 89.28%) to vancomycin, tetracycline, streptomycin, or nalidixic acid was found. Trimethoprim-sulfamethoxazole resistance was present in 32.14%. Relatively lower incidence of resistance (≤ 8.33%) to gentamicin, chloramphenicol, ampicillin, and ceftriaxone was observed. Concurrent resistance to at least 4 antibiotics was detected in 92.85% of the isolates. Cls1 integrons and Int1 were positive in 80.95% and 95.23% of the isolates, respectively. However, Int1 alone was detected in 15.47% (n = 13). In conclusion, the high prevalence of Salmonella spp. in chicken meat may pose a potential public health risk, and the presence of antibiotic-resistant Salmonella spp. isolate together with Cls1 integron and/or integrase might play an important role in horizontal antibiotic gene transfer.

Listeria monocytogenes in retailed raw chicken meat in Turkey.

The objectives of this study were, to find the prevalence and antimicrobial resistance of L. monocytogenes from a total of 116 chicken meat samples including 50 carcasses and 66 meat parts marketed in Turkey between 2008 and 2009 using immunomagnetic separation (IMS) based cultivation technique, to detect the hlyA gene for the verification of the isolates by PCR, and to identify the genoserotypes of the L. monocytogenes isolates by multiplex PCR assay. In the study, 51 L. monocytogenes colonies were isolated from 34 (29.3%) chicken meat samples (eleven [22.0%] carcasses and 23 [34.8%] pieces of meat) by IMS based cultivation technique and confirmed by PCR. According to the multiplex PCR results, all the 51 isolates were identified as genoserotype IIa (1/2a or 3a). L. monocytogenes isolates were also tested for their susceptibility to eight antibiotic (gentamicin, vancomycin, chloramphenicol, streptomycin, tetracycline, ampicillin, penicillin G, erythromycin) agents using the disk diffusion method. 14 isolates (27.45%) were susceptible to all eight antimicrobials drugs tested and the remaining 37 isolates (72.54%) were resistant to gentamicin (one isolate, 1.96%), vancomycin (four isolates, 7.84%), penicillin G (six isolates, 11.76%), streptomycin (nine isolates, 17.64%; resistant or intermediate), tetracycline (seven isolates, 13.72%) and ampicillin (six isolates, 11.76%). This study showed that antimicrobial resistance is not highly prevalent in L. monocytogenes isolated from chicken carcasses and pieces of meat. The presence of L. monocytogenes in chicken samples suggests an importance of this pathogen in chicken.

Prevalence and molecular characterization of sorbitol fermenting and non-fermenting Escherichia coli O157:H7(+)/H7(-) isolated from cattle at slaughterhouse and slaughterhouse wastewater.

The prevalence and seasonal distribution of E. coli O157:H7(+)/H7(-) in an array of aged cattle at slaughter and its dissemination with slaughterhouse wastewater over a two year period in Turkey were investigated. For this purpose, a total of 720 samples (240 rectoanal mucosal swap [RAMS], 240 carcass sponge and 240 bile samples) of 240 cattle categorized according to age, gender, breed and sampling site were collected along with additional 24 wastewater samples and were subjected to immunomagnetic separation based cultivation technique to efficiently isolate E. coli O157 from the background flora. Identification (rfbEO157, fliCh7), detection of major virulence factors (stx1, stx2, eaeA, hly, lpfA1-3 and espA), intimin variants (eae-α1, eae-α2, eae-ß, eae-ß1, eae-ß2, eae-γ1 and eae-γ2/θ) and shiga toxin variants (stx1c, stx1d, stx2c, stx2d, stx2e, stx2f and stx2g) of all the isolates were assessed by PCR. From 10 (4.2%) of RAMS and 11 (4.6%) of carcass sponge samples and 5 (20.8%) of slaughterhouse wastewater samples, a total of 102 colonies (99 sorbitol negative and 3 sorbitol positive) were isolated. Overall, 17 (7.1%) and 15 (6.3%) of 240 sampled cattle were shown to harbor E. coli O157 and E. coli O157:H7, respectively either in their RAMS or carcass sponge samples analyzed. Statistically significant differences between categories; season, age, gender and breed of cattle were not observed (p>0.05). None of the isolated E. coli O157:H7(+)/H7(-) strains harbored any of the investigated intimin types other than eaeγ1 or shiga toxin variants stx1d, stx2e, stx2f or stx2g while all were lpfA1-3(+) except 5 E. coli O157:H7(-) strains. Intimin variant eaeγ1 and shiga toxin 1 variant stx1c were detected from all of the eaeA(+) (97/102, 95.1%) and stx1(+) (32/102, 31.3%) strains, respectively while from stx2(+) (80/102, 78.4%) isolates, both stx2c (68/80, 85.0%) and stx2d (12/80, 15.0%) variants were determined. In the last decade, prevalence of E. coli O157:H7 has an increasing trend in cattle. Slaughterhouses are the significant sources of environmental contamination with E. coli O157:H7. Isolation and molecular characterization of sorbitol fermenting E. coli O157:H7 are a novel finding and may lead to a revision of reference isolation procedure of E. coli O157:H7 in future.

Serotype distribution of Salmonella isolates from turkey ground meat and meat parts.

The aim of the study was to find out the serotype distribution of 169 Salmonella colonies recovered from 112 Salmonella positive ground turkey (115 colonies) and 52 turkey meat parts (54 colonies). Out of 15 Salmonella serotypes: S. Corvallis, S. Kentucky, S. Bredeney, S. Virchow, S. Saintpaul and S. Agona were identified as the predominant serovars at the rates of 27%, 13%, 12%, 12%, 11%, and 10%, respectively. Other serotypes were below 6% of the total isolates. All S. Kentucky and S. Virchow and most of the S. Corvallis (39/46) and S. Heidelberg (9/9) serotypes were recovered from ground turkey. The results indicate that turkey ground meat and meat parts were contaminated with quite distinct Salmonella serotypes. This is the first study reporting Salmonella serotype distribution in turkey meat and S. Corvallis as predominant serotype in poultry meat in Turkey.

Relation between serotype distribution and antibiotic resistance profiles of Listeria monocytogenes isolated from ground turkey.

The objectives of this study were to determine the serotype distribution of Listeria monocytogenes isolated from ground turkey using a multiplex PCR assay and to determine antimicrobial resistance profiles of the isolates using the disc diffusion method. Of 78 isolates, 35 (44.9%), 29 (37.2%), 7 (9.0%), and 7 (9.0%) were identified as serotypes 1/2a (or 3a), 4b (or 4d or 4e), 1/2b (or 3b), and 1/2c (or 3c), respectively. Overall, 63 isolates (80.8%) were resistant to penicillin G, and 53 (67.9%) were resistant to ampicillin. All 1/2c (or 3c) serotype isolates were resistant to penicillin G and ampicillin, and all 1/2b (or 3b) serotype isolates were resistant to penicillin G. In addition, 91.4% (32 of 35) of 1/2a (or 3a), 57.1% (4 of 7) of 1/2b (or 3b), and 37.9% (11 of 29) of 4b (or 4d or 4e) serotype isolates were resistant to ampicillin, and 85.7% (30 of 35) of 1/2a (or 3a) and 65.5% (19 of 29) of 4b (or 4d or 4e) serotype isolates were resistant to penicillin G. In conclusion, most of the L. monocytogenes isolates identified were serotype 1/2a (or 3a) and 4b (or 4d or 4e). Serotype 1/2c (or 3c) isolates were highly resistant to antibiotics compared with isolates of serotypes 1/2a (or 3a), 1/2b (or 3b), and 4b (or 4d or 4e). Increasing resistance of L. monocytogenes to ampicillin and penicillin is an especially serious concern for public health because of the common use of these antibiotics in treatment of human listeriosis cases.

H-NS controls metabolism and stress tolerance in Escherichia coli O157:H7 that influence mouse passage.

BACKGROUND: H-NS is a DNA-binding protein with central roles in gene regulation and nucleoid structuring in Escherichia coli. There are over 60 genes that are influenced by H-NS many of which are involved in metabolism. To determine the significance of H-NS-regulated genes in metabolism and stress tolerance, an hns mutant of E. coli O157:H7 was generated (hns::nptI, FRIK47001P) and its growth, metabolism, and gastrointestinal passage compared to the parent strain (43895) and strain FRIK47001P harboring pSC0061 which contains a functional hns and 90-bp upstream of the open-reading frame. RESULTS: The hns mutant grew slower and was non-motile in comparison to the parent strain. Carbon and nitrogen metabolism was significantly altered in the hns mutant, which was incapable of utilizing 42 carbon, and 19 nitrogen sources that the parent strain metabolized. Among the non-metabolized substrates were several amino acids, organic acids, and key metabolic intermediates (i.e., pyruvate) that limit carbon acquisition and energy generation. Growth studies determined that the parent strain grew in LB containing 14 to 15% bile or bile salts, while the hns mutant grew in 6.5 and 9% of these compounds, respectively. Conversely, log-phase cells of the hns mutant were significantly (p < 0.05) more acid tolerant than the parent strain and hns mutant complemented with pSC0061. In mouse passage studies, the parent strain was recovered at a higher frequency (p < 0.01) than the hns mutant regardless of whether log- or stationary-phase phase cells were orally administered. CONCLUSION: These results demonstrate that H-NS is a powerful regulator of carbon and nitrogen metabolism as well as tolerance to bile salts. It is likely that the metabolic impairments and/or the reduced bile tolerance of the E. coli O157:H7 hns mutant decreased its ability to survive passage through mice. Collectively, these results expand the influence of H-NS on carbon and nitrogen metabolism and highlight its role in the ability of O157:H7 strains to respond to changing nutrients and conditions encountered in the environment and its hosts.