Characteristics of Escherichia coli from raw vegetables at a retail market in the Czech Republic

A large epidemic caused by shigatoxigenic Escherichia coli (E. coli) in spring 2011 in Germany resulted in reduction of trust in the health safety of raw vegetables and sprouted seeds. This study focused on the detection and characterization of E. coli in raw vegetables and sprouted seeds sold in the Czech Republic. Out of 91 samples, 24 (26.4%) were positive for the presence of E. coli. Resistance to antimicrobial agents was determined by the disk diffusion method and E-test. Polymerase chain reaction was used for the detection of selected genes encoding virulence — eaeA, hly, stx₁, and stx₂ and genes encoding resistance to tetracycline — tet(A), tet(B), tet(C), and tet(G) and to β-lactams — bla_TEM, bla_SHV, and bla_CTX. The bla_TEM gene was detected in two isolates, the tet(B) gene in three and tet(A) in one isolate. No hly, stx₁, or stx₂ genes were present, but the eaeA gene was found in three (11.1%) isolates from imported vegetables. These isolates can be considered as potentially enteropathogenic. Results of this study show that raw vegetables and sprouted seeds sold in the retail market can represent a potential risk for consumers.     

Antibiotic resistance in non-enterococcal lactic acid bacteria and bifidobacteria

Over the last 50 years, human life expectancy and quality of life have increased dramatically due to improvements in nutrition and the use of antibiotics in the fight against infectious diseases. However, the heyday of antibiotic treatment is on the wane due to the appearance and spread of resistance among harmful microorganisms. At present, there is great concern that commensal bacterial populations from food and the gastrointestinal tract (GIT) of humans and animals, such as lactic acid bacteria (LAB) and bifidobacteria, could act as a reservoir for antibiotic resistance genes. Resistances could ultimately be transferred to human pathogenic and opportunistic bacteria hampering the treatment of infections. LAB species have traditionally been used as starter cultures in the production of fermented feed and foodstuffs. Further, LAB and bifidobacteria are normal inhabitants of the GIT where they are known to exert health-promoting effects, and selected strains are currently been used as probiotics. Antibiotic resistance genes carried by LAB and bifidobacteria can be transferred to human pathogenic bacteria either during food manufacture or during passage through the GIT. The aim of this review is to address well-stated and recent knowledge on antibiotic resistance in typical LAB and bifidobacteria species. Therefore, the commonest antibiotic resistance profiles, the distinction between intrinsic and atypical resistances, and some of the genetic determinants already discovered will all be discussed.     

Presence and potential for horizontal transfer of antibiotic resistance in oxidase-positive bacteria populating raw salad vegetables

To assess whether domestically grown fresh salad vegetables constitute a possible reservoir of antibiotic resistance for Canadian consumers, aerobic bacteria capable of forming colonies at 30 degrees C on nutrient-limited media were recovered from a single sampling of Romaine lettuce, Savoy spinach and alfalfa sprouts, then examined for their susceptibility to ten antibiotics and the carriage of potentially mobile R-plasmids and integrons. Of the 140 isolates resistant to one or more antibiotic, 93.5 and 90.0% were resistant to ampicillin and cephalothin; 35.7% to chloramphenicol, 10.0% to streptomycin, 4.2% to nalidixic acid, 4.2% to kanamycin, and 2.8% to gentamicin. Gram-positive isolates accounted for less than 4% of the antibiotic resistant strains. A small portion (23.1%) of the predominant oxidase-positive, gram-negative isolates was resistant to two or more antimicrobials. Members of the Pseudomonas fluorescens/putida complex were most prevalent among the 34 resistant strains identified. Sphingobacterium spp. and Acinetobacter baumanni also were detected. Ten of 52 resistant strains carried plasmids, 3 of which were self-transmissible and bore resistance to ampicillin and kanamycin. Eighteen of 48 gave PCR evidence for integron DNA. Class 2 type integrons were the most prevalent, followed by class 1. We conclude that the foods examined here carry antibiotic resistant bacteria at the retail level. Further, our determination that resistant strains contain integron-specific DNA sequences and self-transmissible R-plasmids indicates their potential to influence the pool of antibiotic resistance in humans via lateral gene transfer subsequent to ingestion.     

Antimicrobial resistance and R-factor transfer of salmonellae isolated from chicken carcasses in Greek hospitals

A total of 62 salmonellae, belonging to six different serotypes, were isolated from 60 out of 87 (69.0%) chicken carcasses delivered to hospitals of Thesssaloniki, Greece. Salmonella enteritidis, Salmonella anatum and Salmonella bredeney were the most prevalent serovars. Isolates were examined for antibiotic resistance patterns and R-determinants. Resistance to at least one antibiotic was observed in 36 (58.1%) of them and 18 different resistant profiles were recorded. Nitrofurantoin-resistance was the most common (29.0%), followed by spectinomycin (21.0%), ampicillin (19.4%) and ticarcillin (19.4%). Fourteen (38.9%) of the resistant isolates possessed R-factors and resistance to ampicillin, ticarcillin, trimethoprim and kanamycin was easily self-transferable. However, nitrofurantoin- and spectinomycin-resistance although prevailing, was not found transferable even after mobilization. The high incidence of antibiotic resistant salmonellae among chicken carcasses in our hospital setting suggests the need for public health interventions and possible withdrawal of drug selective pressure.     

Antibiotic resistance of coagulase-negative staphylococci associated with food and used in starter cultures

The resistance of 330 coagulase-negative staphylococci (CNS) associated with food or used in starter cultures and belonging to the species Staphylococcus carnosus, Staphylococcus condimenti, Staphylococcus piscifermentans, Staphylococcus equorum, Staphylococcus succinus and Staphylococcus xylosus, against 21 antibiotics was determined using the disk diffusion method. The incidence and number of resistances was found to be species and source of isolation dependent. Most strains of S. equorum (63%), S. succinus (90%) and S. xylosus (95%) exhibited resistances against up to seven antibiotics, whereas only few strains of S. carnosus (12%) and S. piscifermentans (27%) were antibiotic resistant. Resistances to lincomycin, penicillin, fusidic acid, oxacillin, ampicillin and tetracycline were predominant. Among strains of S. xylosus, the incidence of resistance ranged from 22% for tetracycline up to 69% for penicillin. Concerning the source of isolation, resistances were often determined in strains of S. equorum, S. succinus and S. xylosus isolated from cheese (87%) and sausage (83%), and strains of S. xylosus obtained from meat starter cultures (93%). Remarkably, all CNS were sensitive to the clinically important antibiotics chloramphenicol, clindamycin, cotrimoxazol, gentamicin, kanamycin, linezolid, neomycin, streptomycin, synercid and vancomycin. The phenotypic resistances to ss-lactam antibiotics, lincomycin and tetracycline were verified by PCR amplification and could be traced back to the genes blaZ, lnuA and tetK, respectively. This study permitted a comprehensive insight into the incidence of antibiotic resistances in food-associated CNS.     

Antibiotic resistance genes and identification of staphylococci collected from the production chain of swine meat commodities

Staphylococci harbouring antibiotic resistance (AR) genes may represent a hazard for human health and, as other resistant food-related bacteria, they contribute to the spread of AR. In this study, we isolated resistant staphylococci from an entire swine production chain and investigated the occurrence of 11 genes [aac(6')Ie-aph(2')Ia, blaZ, mecA, vanA, vanB, ermA, ermB, ermC, tet(M), tet(O) and tet(K)] encoding resistance to some antibiotics largely used in clinical practice. The 66 resistant staphylococcal isolates were identified as Staphylococcus epidermidis (27 isolates), Staphylococcus aureus (12), Staphylococcus xylosus (12), Staphylococcus simulans (5), Staphylococcus pasteuri (4), Staphylococcus carnosus (3), Staphylococcus lentus (2) and Staphylococcus sciuri (1). Specific-PCR detection of AR genes showed the prevalence of the tet(K) gene in most of the isolates (89.4%), followed by tet(M) and ermC (about 75%); mecA was detected in more than half of S. aureus and S. epidermidis isolates. The genes vanA and vanB were not retrieved. It was found that a high proportion of coagulase-positive and -negative isolates are multidrug-resistant and some of them carry up to six AR genes. Our findings show that the swine production chain is a source of antibiotic-resistant staphylococci suggesting the importance of resistance surveillance in the food production environment.     

市售即食食品中非致病菌的耐药性及耐药基因转移的研究

目的 了解西安市即食食品中的非致病菌的耐药性分布、耐药程度及耐药基因的转移情况,分析潜在的食品安全问题.方法 对西安市的几家大型超市零售的熟肉、果盘、凉拌菜3大类即食食品样品进行食源性非致病菌的红霉素和氨苄青霉素耐药性分析、耐药基因转移、菌株16 S rRNA序列测序鉴定等.结果 42.5％的分离株具有不同程度的红霉素耐药性,47.5％的分离株具有不同程度的氨苄青霉素耐药性,21.2％的分离株同时具有2种抗生素耐药性;11.3％的红霉素耐药株耐药性高达160 mg/L,6.3％的氨苄青霉素耐药株耐药性高达500 mg/L.随机挑选的部分耐药株在无抗生素培养基中连续传代1000代以后,60％的氨苄青霉素耐药株和45％的红霉素耐药株的耐药性消失.部分耐药株的耐药基因可水平转移至非耐药株中.这些耐药株多为肠球菌、葡萄球菌、芽孢杆菌、假单胞菌等.结论 即食食品中的非致病菌有潜在的风险,耐药基因在细菌间的转移可能对人体敏感肠道菌群和致病菌有一定影响.     

A note on the incidence and antibiotic resistance of Staphylococcus aureus isolated from meat and chicken samples

One hundred and fifty samples of raw calf/lamb meat samples (mince and chunks) and chicken parts (giblets, carcass) were analysed for the presence of Staphylococcus aureus. Eighty S. aureus strains were isolated and identified. Resistance of the strains to methicillin and other antibiotics was determined by the Kirby-Bauer disc diffusion test. The overall methicillin resistance rate for S. aureus was 67.5%. Of S. aureus strains, 87.5% were resistant to bacitracin. A high prevalence of penicillin G resistance was detected for S. aureus (53.8%). Few of the strains were resistant to erythromycin (7.5%). All strains were susceptible to vancomycin, sulbactam–ampicillin, ciprofloxacin and cefaperazone–sulbactam. This study confirmed the presence of S. aureus, especially antibiotic-resistant strains, in the foods examined, indicating poor sanitary conditions during processing which may create a health risk for consumers.     

Identification and functional traits of lactic acid bacteria isolated from Ciauscolo salami produced in Central Italy

Lactic acid bacteria (LAB) from Ciauscolo salami produced in Marche Region of Central Italy, and LAB strains belonging to our laboratory collection were examined for their capability to survive at low pH and bile, to adhere to Caco-2 cells, and for antibiotic resistance. LAB from Ciauscolo were identified by ARDRA and RAPD-PCR. Our study showed that all LAB strains had good adaptation to gastric juice and moderate tolerance to bile. The adhesiveness was variable among strains but significantly lower in LAB from food. Antibiotic resistance was broadly spread among food strains, with level of resistance exceeding 15% for all the antibiotics tested. The resistance determinants erm(B) and tet(M) were found in nine strains of food origin (21.4%) while tet(L) in one strain of our collection (5%). Our work suggests that fermented foods are valuable sources of bacterial strains with functional traits of intestinal lactobacilli. These bacteria may be further studied for their use in probiotic applications.     

泡菜中乳酸菌的分离鉴定及体外抗性筛选

从泡菜中分离得到5株乳酸杆菌,经16S rDNA种属分析,其中4株为植物乳杆菌(Lactobacillus plantarum),编号分别为E-6-1、E-1-2、E-6-3、D-2-3,1株为消化乳杆菌(Lactobacillus alimentarius),编号为C-2-1。通过人工胃液和胆盐试验来确定5株菌的体外抗性。结果表明,D-2-3乳酸菌具有较好的抗人工胃液能力,其在pH 3.0的人工胃液处理3h后的存活率高达95.38%,且在1,2,3g/L胆盐中均能较好的生长。说明该菌株具有益生菌潜质。     

Functional properties and safety assessment of lactic acid bacteria isolated from goat colostrum for application in food fermentations

Thirty representative isolates from genotypes obtained after RAPD‐PCR analysis of 191 presumptive LAB isolates from goat colostrum were assessed. They were identified as Leu. mesenteroides, Leu. lactis and Lac. garvieae. Only Lac. garvieae P27M15 showed a high resistance to low pH, while 93.3% of the isolates retained their viability after exposure to bile salts. Leu. lactis V22M11, V42M18 and V23M15 displayed the best technological properties. Lac. garvieae P27M15 and Leu. lactis V22M11, V42M18 and V23M15 were susceptible to 8 of the 13 antibiotics assayed. Therefore, it can be concluded that the isolates P27M15, V22M11, V42M18 and V23M15, showing potentially important properties, could be valuable for practical applications in biotechnological processes.     

Evaluation of antibacterial resistance in Vibrio strains isolated from imported seafood and Italian aquaculture settings

Ninety-two Vibrio strains isolated over a period of 9 years from different sources (national and imported fishery products, shellfish, seawater from aquaculture settings, etc.) and belonging mostly to two species relevant for human health and fish pathology, V. parahaemolyticus and V. alginolyticus, were tested for resistance to different antibacterials using the standard disk diffusion test (five antibacterials: ampicillin, sulfamethoxazole, tetracycline, trimethoprim/sulfamethoxazole and chloramphenicol) and the Minimum Inhibitory Concentrations method (six antibacterial families: β-lactams, tetracyclines, chloramphenicol, aminoglycosides, quinolones, trimethoprim/sulfamethoxazole) providing consistent results. The analysis performed by standard disk diffusion test showed resistance to ampicillin in 82% of the strains, whereas 7% were resistant to sulfamethoxazole, 3% to tetracycline, and 1% to trimethoprim/sulfamethoxazole; no strain was found to be resistant to chloramphenicol. The results obtained by the MIC tests, because of the absence of established breakpoints for some antibacterials, were not readily interpretable for all substances, but showed no statistically significant difference to the results obtained by the standard disk diffusion test. Beside these results, high MIC₉₀ values (128 μg ml⁻¹ and above) were obtained for kanamycin and streptomycin, indicating that high concentrations of these antibacterials were required to inhibit the growth of the strains. Strains exhibiting resistance or intermediate resistance to two or more antibacterials represented 15% of the total and included, beside resistance to ampicillin, resistance to sulfamethoxazole or, for V. parahaemolyticus isolates, tetracycline. Interestingly, differences in the mechanism of resistance to β-lactams were detected between the species V. parahaemolyticus and V. alginolyticus.     

2016-2018年北京市腹泻患者弯曲菌耐药性与脉冲场凝胶电泳分子分型的研究

目的 分析北京市腹泻患者弯曲菌(Campylobacter spp.)分离株的抗生素敏感性和脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)分子分型特征.方法 2016—2018年,对北京市19家医院肠道门诊分离的弯曲菌采用琼脂稀释法进行抗生素敏感性检测,按照PulseNet...     

Occurrence,heat and antibiotic resistance profile of Bacillus cereus isolated from raw cow and processed milk in Mezam Division,Cameroon

Bacillus cereus is the aetiologic agent of two distinct forms of food poisoning: the diarrhoeal and emetic syndromes. Little data exist on the prevalence of B. cereus in raw milk and milk products sold in Cameroonian towns. This study was aimed at investigating the occurrence, heat and antibiotic resistance of B. cereus isolated from raw milk and selected milk products in Mezam division, Cameroon. Bacillus cereus was isolated by inoculating samples onto mannitol‐egg yolk‐polymyxin B agar. Isolates were characterised morphologically and biochemically. The occurrence of B. cereus in raw milk (8.22%) was less than that in milk powder (13.33%). Bacillus cereus was not isolated from fermented milk. There was no significant difference (P < 0.05) between the B. cereus load in raw milk (2.6 × 10³ cfu/mL) and milk powder (3.0 × 10² cfu/mL). All the isolates showed haemolysin activity and were sensitive to tetracycline, gentamicin, chloramphenicol and nalidixic acid, but resistant to penicillin, ampicillin and trimethoprim/sulphamethoxazole. The detection of drug‐resistant, haemolysin‐positive isolates should serve as a warning for an impending health hazard following consumption of untreated milk. Heat resistance of isolates was assessed by determining the decimal reduction time; D‐value (time to inactivate 90% of the B. cereus spores); and the heat sensitivity, z (temperature increase leads to a tenfold reduction in the D‐value). The values for D₁₀₀ ranged from 0.5 to 3.5 min, and z‐values ranged from 10.0 to 32.6 °C. These results could be used in the dairy industry to evaluate the importance of heat treatment on B. cereus inactivation and calculation of process efficiency.     

Characterization of Listeria monocytogenes isolates from lactating dairy cows in a Wisconsin farm: Antibiotic resistance,mammalian cell infection,and effects on the fecal microbiota

Listeria monocytogenes is an invasive foodborne pathogen that is ubiquitously present in the dairy farm environment. Although cattle are a reservoir of L. monocytogenes, most adult animals do not exhibit clinical symptoms, suggesting a homeostasis between this pathogen and the bovine gastrointestinal ecosystem. Nevertheless, substantial prevalence of L. monocytogenes fecal shedding by dairy cattle has been reported in many studies, posing threats of transmission within the herd and contamination of the human food supply. Accordingly, understanding the L. monocytogenes ecology within the bovine gastrointestinal tract is important to prevent clinical illness in the animal host, reduce transmission, and guide intervention strategies. In this study, we conducted a longitudinal sampling of fecal samples from 20 lactating dairy cows in one Wisconsin farm over a 29-d period and found a strikingly high incidence of L. monocytogenes shedding, in 90% of sampled animals. The L. monocytogenes isolates were genetically diverse, representing all common serotypes previously identified from cattle. Additionally, most tested isolates were resistant to ampicillin, and a few were also resistant to gentamicin or trimethoprim/sulfamethoxazole. Most isolates effectively infected human epithelial cells (Caco-2) and murine fibroblasts (L2), suggesting that they are all capable of causing systemic infection if the intestinal barrier is breached. Finally, we investigated the effects of L. monocytogenes colonization on the gastrointestinal tract microbiota by analyzing the fecal bacterial communities of some shedding and nonshedding cows. Whereas L. monocytogenes did not affect the α and β diversity of tested animals, a subset of shedding cows exhibited different abundances of certain operational taxonomic units within the Bacteroidetes and Firmicutes phyla compared with nonshedding cows. Overall, our findings highlight the threat of antibiotic resistance among some L. monocytogenes isolates, emphasize the need for a strain-specific approach in listeriosis treatment, and suggest the potential negative influence of subclinical L. monocytogenes carriage on animal gut health.     

Analysis of antibiotic resistance patterns and detection of mecA gene in Staphylococcus aureus isolated from packaged hamburger

Presence of Staphylococcus aureus, antibiotic resistance pattern and PCR detection of mecA gene in isolated strains were investigated in total of 256 packaged hamburgers in Iran-Tehran. For this purpose we used standard disk-diffusion method and sensitive and specific PCR technique, respectively. Results showed that 25% of samples were positive for S. aureus. Resistance to meticillin, erythromycin, penicillin G, cefazolin, ciprofloxasin, vacomycin and amoxiclave was determined 89%, 20.3%, 18.7%, 15.6%, 14%, 26.6% and 12.5%, respectively. According to the obtained results from PCR analysis of methicillin-resistant S. aureus (MRSA), mecA gene was present in 100% of the resistant isolates, 0% of intermediate-resistance isolates and 25% of susceptible isolates. The results obtained from PCR detection of mecA gene showed high correlation with standard disk diffusion test.     

Antibiotic resistance in gut bacteria from dairy calves: a dose response to the level of antibiotics fed in milk

Dairy calves are commonly fed milk from cows treated with antibiotics. The concentration of beta-lactam antibiotic residues found in milk from treated cows was used to determine the range of concentrations of penicillin used in a dose-regulated experiment. Thirty-one Holstein calves were randomly assigned to milk with penicillin G added at concentrations of 0, 6.25, 12.5, 25, and 50 microl/kg. Fecal swabs were taken from each calf twice weekly. Resistance to penicillin was tested by measuring the zone of inhibition in bacterial growth around a disk impregnated with the antibiotic. Inhibition was greatest for bacteria from calves fed milk with no penicillin (2.89 +/- 0.14 mm), and declined as the penicillin dose provided in the milk increased (to a low of 0.70 +/- 0.10 for the 50 microl/kg treatment group). In conclusion, resistance of gut bacteria to antibiotics increases with increasing concentrations of penicillin in the milk fed to dairy calves.     

Occurrence, virulence genes and antibiotic resistance of Escherichia coli O157 isolated from raw bovine, caprine and ovine milk in Greece

The examination of 2005 raw bovine (n = 950), caprine (n = 460) and ovine (n = 595) bulk milk samples collected throughout several regions in Greece for the presence of Escherichia coli serogroup O157 resulted in the isolation of 29 strains (1.4%) of which 21 were isolated from bovine (2.2%), 3 from caprine (0.7%) and 5 from ovine (0.8%) milk. Out of the 29 E. coli O157 isolates, only 12 (41.4%) could be classified as Shiga-toxigenic based on immunoassay and PCR results. All 12 Shiga-toxigenic E. coli serogroup O157 isolates belonged to the E. coli O157:H7 serotype. All except one of the 12 Shiga-toxin positive isolates were stx₂-positive, five of which were also stx₁-positive. The remaining isolate was positive only for the stx₁ gene. All stx-positive isolates (whether positive for stx₁, stx₂ or stx₁ and stx₂) were also PCR-positive for the eae and ehxA genes. The remaining 17 E. coli O157 isolates (58.6%) were negative for the presence of the H7 flagellar gene by PCR, tested negative for Shiga-toxin production both by immunoassay and PCR, and among these, only four and three strains were PCR-positive for the eae and ehxA genes, respectively. All 29 E. coli O157 isolates displayed resistance to a wide range of antimicrobials, with the stx-positive isolates being, on average, resistant to a higher number of antibiotics than those which were stx-negative.