Species distribution,antibiotic resistance and virulence traits in enterococci from meat in Tunisia

Antimicrobial resistance and the mechanisms implicated were studied in 119 enterococci from 105 meat samples from Tunisian markets. Almost 24.5% of recovered enterococci showed resistance against four or more antimicrobial agents and these isolates were identified to the species level. Enterococcus faecalis was the most prevalent species (41%). High percentages of erythromycin and tetracycline resistances were found among our isolates, and lower percentages were identified to aminoglycosides, ciprofloxacin and chloramphenicol. All tetracycline-resistant isolates carried the tet(M) and/or tet(L) genes. The erm(B) gene was detected in 78.5% of erythromycin-resistant isolates, ant(6)-Ia gene in 58.8% of streptomycin-resistant isolates, and cat(A) gene in one chloramphenicol-resistant isolate. Forty-eight isolates carried the gelE gene and exhibited gelatinase activity. The hyl and esp genes were detected in one and three Enterococcus faecium isolates, respectively. Streptomycin-resistant isolates showed a high genetic diversity by PFGE and MLST. Meat might play a role in the spread through the food chain of enterococci with these virulence and resistance characteristics to humans.     

Characterization of Streptococcus lutetiensis isolated from clinical mastitis of dairy cows

Streptococcus lutetiensis, previously termed Streptococcus bovis type II/1, has rarely been associated with bovine mastitis. The objectives of this work were to characterize the molecular diversity, antimicrobial resistance profiles, virulence genes of Strep. lutetiensis (n = 37) isolated from bovine clinical mastitis, as well as its pathogenic effects in a murine mastitis model. Genetic relationships of isolates were determined by random amplified polymorphic DNA (RAPD)-PCR, virulence genes were detected by PCR. Antimicrobial susceptibility testing was carried out by broth microdilution technique. The pathogenic effects of Strep. lutetiensis were studied with 2 infection models: bovine mammary epithelial cells cultured in vitro and murine mammary infection in vivo. Streptococcus lutetiensis isolates were clustered into 5 RAPD-types (A–E), with a dominant type A representing 84% of isolates. Eighteen (49%), 16 (43%), and 9 (24%) isolates were resistant to ceftiofur, tetracycline, and erythromycin, respectively. Prevalence of multidrug resistance (resistant to ≥3 classes of antimicrobials) was 24% (9/37). The most prevalent virulence genes were bca (100%), speG (100%), hly (97%), scpB (95%), and ssa (95%). There was no difference between isolates from mild and moderate cases of bovine mastitis in prevalence of virulence genes. Streptococcus lutetiensis rapidly adhered to and subsequently invaded (1 and 3 h after infection, respectively) bovine mammary epithelial cells, resulting in elevated lactate dehydrogenase release (4 h after infection). Edema and hyperemia were observed in challenged mammary glands and bacteria were consistently isolated at 12, 24, and 48 h after infection. In addition, numerous neutrophils migrated into gland alveoli and interstitium of infected mammary tissue. We concluded that Strep. lutetiensis had potential to spread within a dairy herd and good adaptive ability in bovine mammary cells or tissue, which are generally characteristics of a contagious mastitis pathogen.     

Prevalence and characterization of antibiotic resistant Enterococcus faecalis in French cheeses

Prevalence of enterococci and antibiotic resistance profiles of Enterococcus faecalis was analyzed in 126 French cheeses from retail stores. Forty-four percent of pasteurized or thermised-milk cheeses, and up to 92% of raw-milk cheeses contained detectable enterococci. A total of 337 antibiotic resistant enterococci were isolated in 29% and 60% of pasteurized-milk and raw-milk cheeses, respectively. E. faecalis was the predominant antibiotic resistant species recovered (81%), followed by Enterococcus faecium (13%), and Enterococcus durans (6%). The most prevalent antibiotic resistances were tetracycline (Tet) and minocycline (Min), followed by erythromycin (Ery), kanamycin (Kan) and chloramphenicol (Cm). The most common multiple antibiotic resistance phenotype was Cm Ery Kan Min Tet. The occurrence of antibiotic genes, as searched by PCR, was 100 % for aph3′IIIa, 96 % for ermB, 90 % for tetM and 80 % for catA in isolates resistant to Kan, Ery, Tet or Cm, respectively. MLST analysis of 30 multidrug resistant E. faecalis revealed that ST19, CC21, CC25 and CC55 isolates were the most common in cheeses. In conclusion, as in many other European countries, French cheeses do contain enterococci with multiple antibiotics resistances. However, low occurrence of high-level gentamicin resistant or sulfamethoxazole/trimethoprim-resistant enterococci and absence of vancomycin- or ampicillin- resistant enterococci indicate that cheeses cannot be considered as a major reservoir for nosocomial multi-drug resistant enterococci.     

Minimum inhibitory concentrations and disk diffusion zone diameter for selected antibiotics against streptococci isolated from bovine intramammary infections

The antimicrobial susceptibility to 11 antibiotics was determined of 358 streptococcal isolates representing six species was determined. All isolates were from milk samples obtained from clinical or subclinical cases of bovine mastitis. Agar disk diffusion zone diameters and minimum inhibitory concentrations were obtained for all isolates. Of the antibiotics tested that are available for mastitis therapy, cephalothin, penicillin, ampicillin, novobiocin, and erythromycin demonstrated acceptable activity against nonenterococcal streptococci, but only penicillin and ampicillin demonstrated moderate activity against Enterococcus faecalis. Enterococcus faecalis demonstrated substantial resistance to cephalothin, erythromycin, gentamicin, streptomycin, and tetracycline, while the nonenterococcal streptococci demonstrated substantial resistance to gentamicin, streptomycin, and tetracycline. Streptococcus uberis demonstrated greater antibiotic resistance than did Streptococcus dysgalactiae, while Aerococcus viridans demonstrated the greatest overall resistance of the nonenterococcal species.     

Minimum inhibitory concentrations of Staphylococcus aureus recovered from clinical and subclinical cases of bovine mastitis

Antimicrobials are often used for treatment of bovine mastitis and the possibility of selection for resistant bacteria must be considered. The objectives of this study were to determine the minimum inhibitory concentration of Staphylococcus aureus recovered from cases of clinical and subclinical bovine mastitis, and to determine the prevalence of multidrug resistance in this population. Milk samples were collected from cows on commercial dairy herds (n=13), including quarters (n=1,574) of cows with subclinical mastitis cases, and cows experiencing clinical mastitis cases (n=608). Selected Staph. aureus isolates, obtained from clinical (n=58) and subclinical (n=58) mastitis cases, were used to determine minimum inhibitory concentrations of 12 selected antimicrobials. Of Staph. aureus isolates tested, 87 (75%) did not exhibit resistance to any antimicrobial, 28 (24.1%) exhibited resistance to 1 (n=21) or 2 (n=7) classes of antimicrobials, and 1 (0.9%) exhibited multidrug resistance. All Staph. aureus (clinical and subclinical cases) were inhibited by the range of concentrations tested for ceftiofur and oxacillin. Moreover, no isolates obtained from clinical mastitis cases exhibited resistance to cephalothin, penicillin-novobiocin, or sulfadimethoxine. Of isolates, 3 exhibited resistance to enrofloxacin. Of isolates exhibiting resistance to more than 1 antimicrobial, independent of antimicrobial class, the combination of erythromycin and tetracycline, and ampicillin and penicillin accounted for the majority of resistance. Of isolates tested, 19% were resistant to tetracycline and 14% were resistant to penicillin. Survival curves of Staph. aureus relative to minimum inhibitory concentration demonstrated heterogeneity among case types for ceftiofur, cephalothin, and erythromycin. Multidrug resistance was identified in only 1 isolate obtained from a single farm.     

Short communication: Prevalence and antibiotic resistance of Staphylococcus aureus isolated from bovine clinical mastitis

The aims of this study were to determine the prevalence and antibiotic resistance of Staphylococcus aureus isolated from bovine clinical mastitis in Varamin, Tehran Province, Iran. All of the isolated Staph. aureus were identified by morphology and culture and confirmed using the API Staph identification system (bioMérieux, Marcy-l’Étoile, France). Antibiotic resistance genes were detected by PCR with oligonucleotide primers specific for each gene. Staphylococcus aureus was recovered from 43 of 207 (20.1%) bovine clinical milk samples. Using disk diffusion, methicillin-resistant Staph. aureus was detected in 5 of 43 (11.6%) samples. The pathogen showed high resistance against penicillin G (86%) and tetracycline (76.7%). The blaZ (penicillin) (86%), tetM (tetracycline), and ermC (erythromycin) genes (39.5% each) were the most prevalent antibiotic resistance genes. The findings of this study are useful for designing specific control programs for bovine clinical mastitis caused by Staph. aureus in this region of Iran.     

Monitoring of radical scavenging compounds from oxidized lipids (RSOL) by 2,4-dinitrophenylhydrazine (DNPH) derivatization in thermally oxidized linoleic acid and lard systems

From February to October 2011, 209 samples of retail raw meat and fishery products were randomly obtained from 41 grocery stores in Korea and cultured for the presence of Enterococcus spp. Ninety-six enterococcal isolates were recovered from 76 samples, with contamination rates ranging from 18.5% in fishery product samples to 43.9% in chicken samples. Antimicrobial disk susceptibility testing was conducted according to the Clinical and Laboratory Standards Institute protocol. The antibiotic resistance rates of the 96 enterococci isolates were as follows: tetracycline (TE) 77.1%; erythromycin (E) 50%; rifampin (RD) 44.8%; and vancomycin (VAN) 9.4%. Disk diffusion showed that 9 isolates were resistant to vancomycin. The minimum inhibitory concentration (MIC) was ≥32 μg/mL for all 9 isolates, and all were resistant to vancomycin. Among the 9 vancomycin-resistant enterococci (VRE) identified, the vanA gene was carried by 1 Enterococcus durans strain and the vanB gene was carried by 2 Enterococcus faecium, and 1 Enterococcus hirae strains. Further genotyping of the VRE isolates using pulsed-field gel electrophoresis (PFGE) revealed extensive heterogeneity. The VRE were recovered from various animal sources with a particularly high prevalence of E. faecium found in fishery products and chicken.     

Short communication: Detection of antibiotic resistance,mecA, and virulence genes in coagulase-negative Staphylococcus spp. from buffalo milk and the milking environment

The aim of this study was to determinate whether coagulase-negative staphylococci (CNS) from buffalo milk or the milking environment possess virulence factors that are associated with intramammary infections or antimicrobial resistance. Milk samples (n = 320) from 80 lactating buffalo were evaluated for clinical and subclinical mastitis by physical examination, the strip cup test, California Mastitis Test (CMT), and somatic cell count (SCC) over a 4-mo period. In addition, swabs were obtained from the hands of consenting milkers (16), liners (64), and from the mouths (15) and nostrils (15) of buffalo calves. No clinical cases of mastitis were observed; however, CMT together with SCC results indicated that 8 animals had subclinical mastitis. Eighty-four CNS isolates were identified by MALDI-TOF MS and cydB real-time PCR (qPCR) and then evaluated by qPCR for presence of the eta, etb, sea, sec, cna, seb, sei, seq, sem, seg, see, and tst toxin genes, adhesion- and biofilm-associated genes (eno, ebps, fib, fnbA, coa), and the methicillin resistance gene (mecA). Resistance to antibiotics commonly used for mastitis treatment in Brazil was determined using the Kirby-Bauer test. Two strains were positive for the see and eta toxin genes; and mecA (1), eno (27), ebps (10), fnbA (10), and coa (5) genes were also detected. A notable number of isolates were resistant to erythromycin (30), penicillin (26), and cotrimoxazole (18); importantly, 10 vancomycin-resistant isolates were also detected. A smaller number of isolates were resistant to rifampicin (8), oxacillin (7), clindamycin (5), cefepime (4), tetracycline (3), ciprofloxacin (2), and chloramphenicol (1), and none were resistant to gentamicin or ciprofloxacin. Isolates with resistance to 2 (13 isolates), 3 (3), 4 (3), 5 (1), and 6 (1) antibiotics were detected. Taken together, our findings suggest that CNS isolates may not be a significant cause of clinical or even subclinical mastitis in buffaloes, but they may be a reservoir of virulence and antibiotic resistance genes.     

The phylogenetic group,antimicrobial susceptibility,and virulence genes of Escherichia coli from clinical bovine mastitis

Bovine mastitis is still a central problem on dairy farms despite control programs, and Escherichia coli is a crucial pathogen during the development of bovine mastitis. The virulence genes, antimicrobial susceptibility, and mortality of mice infected with different E. coli isolates from bovine mastitis were determined in this study. According to the presence of the specific genes chuA, yjaA, and TspE4.C2, these isolates mainly belonged to 2 different groups: group A (47/79) and group B1 (22/79). The ompC gene was detected in all the isolates, followed by fimH (89.9%), ECs3703 (88.6%), and ompF (73.4%), whereas most of the virulence genes were not detected in these isolates. The results of the antimicrobial susceptibility tests indicated that the isolates were susceptible to the fluoroquinolones and aminoglycosides. An inverse relationship was shown between the expression level of ompF and antimicrobial resistance; additionally, the isolates that were nonsusceptible to at least 4 classes of antimicrobial agents showed a lower mortality to mice in comparison with the susceptible isolates. This study indicated that antibiotic resistance had emerged in E. coli from bovine mastitis in this area, and appropriate measures should be taken to avoid potential threats to humans and other animals.     

Antimicrobial activity of binary combinations of natural and synthetic phenolic antioxidants against Enterococcus faecalis

This study investigated the antimicrobial activity of 3 natural (thymol, carvacrol, and gallic acid) and 2 synthetic [butylated hydroxyanisole (BHA) and octyl gallate] phenolic compounds, individually and in binary combinations, on 4 dairy isolates of Enterococcus faecalis with different virulence factors (β-hemolytic, gelatinase, or trypsin activities; acquired resistance to erythromycin or tetracycline; and natural resistance to gentamicin). A checkerboard technique and a microdilution standardized method were used. All compounds individually tested exhibited antimicrobial activity against E. faecalis, with minimal inhibitory concentrations (MIC) ranging from 30 μg/mL (octyl gallate) to 3,150 μg/mL (gallic acid), although no significant differences were detected among strains to each phenolic compound. Carvacrol in combination with thymol or gallic acid, and gallic acid combined with octyl gallate showed partial synergistic inhibition of all E. faecalis strains. The most effective combinations were thymol + carvacrol and gallic acid + octyl gallate, as the MIC for each of these compounds was reduced by 67 to 75% compared with their respective individual MIC. These results highlight the possibility of using combinations of these phenolic compounds to inhibit the growth of potential virulent or spoilage E. faecalis strains by reducing the total amount of additives used in dairy foods.     

In vitro growth of enterococci of bovine origin in bovine mammary secretions from various stages of lactation

In vitro growth responses of Enterococcus faecium and Enterococcus faecalis were tested in cell-free, fat-free bovine mammary secretions. Mammary secretions were collected during the dry period, and during early, late, and extended lactation. Sixty-three enterococcal isolates from aseptically collected bovine quarter milk samples and bedding samples from a commercial dairy herd were tested. Isolates from bovine quarter milk samples originated from mammary glands with clinical mastitis, cows with composite somatic cell score >4, postpartum milk samples, or from routine milk samples submitted to a mastitis diagnostic laboratory. Source of enterococcal isolates and the species significantly contribute to the ability of organisms to multiply in mammary secretions from various stages of lactation. Isolates collected from milk samples of the commercial herd and isolates from milk submitted to a mastitis diagnostic lab did not display enhanced growth in mammary secretions compared with isolates from bedding. Growth responses of E. faecalis were greater than those for E. faecium in secretions collected during the dry period, late lactation, and extended lactation. Bacterial growth did not differ between enterococcal species in mammary secretion collected from cows in early lactation. Differences in bacterial growth between E. faecalis and E. faecium in mammary secretions may indicate differences between species in susceptibility of mammary glands during the lactation cycle.     

Molecular characterization and clonal diversity of methicillin-susceptible Staphylococcus aureus in milk of cows with mastitis in Brazil

Mastitis is an important disease for the dairy industry worldwide, causing economic losses and reducing milk quality and production. Staphylococcus aureus is a worldwide agent of this intramammary infection, which also causes foodborne diseases. The objective of this study was to determine the frequency of methicillin-susceptible Staphylococcus aureus (MSSA) isolates in milk of mastitis cows in Brazil and to analyze the genetic lineages and the content of antimicrobial resistance genes and virulence factors among these isolates. Fifty-six MSSA isolates were recovered from 1,484 milk samples (positive for the California mastitis test) of 518 cows from 11 different farms in Brazil (representing 51% of total Staph. aureus obtained), and they were further characterized. Methicillin-susceptible Staphylococcus aureus were isolated from 3.7% of California mastitis test-positive tested milk samples and from 6.2% of tested mastitic cows. Methicillin-susceptible Staphylococcus aureus isolates were characterized by spa typing, agr typing, and multilocus sequence typing, and resistance and virulence traits were investigated by PCR. Seven spa types were identified among MSSA (% of isolates): t127 (44.6), t605 (37.5), t002, t1784, t2066 (1.8), and 2 new ones: t10856 (10.7) and t10852 (1.8). Five distinct sequence types (ST) were detected (% of isolates): ST1 (46.4), ST126 (37.5), ST133 (10.7), ST5 (3.6), and a novel ST registered as ST2493 (1.8). Resistances were detected for streptomycin, chloramphenicol, and tetracycline. One strain contained the chloramphenicol resistance gene (fexA; included within transposon Tn558) and 3 strains contained the tetracycline resistance gene [tet(K)]. Methicillin-susceptible Staphylococcus aureus strains were susceptible to most of the antibiotics studied and lacked the virulence genes of Panton-Valentine leukocidin (lukF/S-PV), toxic shock syndrome toxin 1 (tst), exfoliative toxin A (eta), and exfoliative toxin B (etb), as well as the genes of the immune evasion cluster. Methicillin-susceptible Staphylococcus aureus isolates were detected in a relatively low proportion of cows with mastitis (6.2%) and recovered isolates presented high diversity of genetic lineages, with CC1 and CC126 the predominant clonal complexes, and CC133 also being detected. Larger epidemiological studies with molecular characterization of isolates are required to deepen the knowledge on the circulating genetic lineages among the cow population with mastitis.     

Short communication: Antimicrobial resistance and virulence characterization of methicillin-resistant staphylococci isolates from bovine mastitis cases in Portugal

Methicillin-resistant staphylococci (MRS) have already been reported as mastitis agents. Such bacterial species are a public health concern, and the characterization of their antimicrobial resistance and virulence profile is important to better control their dissemination. The present work evaluated the distribution of methicillin-resistance among 204 staphylococci from clinical (n = 50) and subclinical (n = 154) bovine mastitis. The presence ofthe mecA gene was determined by PCR. Phenotypic expression of coagulase, DNase, lipase, gelatinase, hemolytic enzymes, and biofilm production was evaluated. The presence of biofilm-related genes, icaA, icaD, and bap, was also determined. Antimicrobial resistance patterns for aminoglycosides, lincosamides, macrolides, fluoroquinolones, sulphonamides, tetracyclines, and fusidic acid were determined. Nineteen (9.3%) isolates were identified as MRS, and the presence of mecA in these isolates was confirmed by PCR. Virulence factors evaluation revealed that gelatinase was the most frequently detected (94.7%), followed by hemolysins (73.7%) and lipase (68.4%); 84.2% of the MRS isolates produced biofilm and icaA and icaD were detected in almost half of the MRS isolates (52.6%), but all were bap-negative. Resistance against other antimicrobial agents ranged from 0 (fusidic acid, ciprofloxacin, norfloxacin, enrofloxacin) to 100% (nalidixic acid). Resistance to nalidixic acid and nalidixic acid-tetracycline were the most common antimicrobial resistance profiles (31.6%). This study confirms that despite the low prevalence of MRS, isolates frequently express other virulence traits, especially biofilm, that may represent a serious challenge to clinicians.     

Short communication: Characterization of enterotoxin-producing Staphylococcus aureus isolated from mastitic cows

Staphylococcus aureus is not only a common cause of bovine mastitis, but also an agent of food poisoning in humans. In an attempt to determine whether staphylococci causing bovine mastitis could also cause food poisoning, 60 isolates of presumed S. aureus were isolated in the period between March and August 2017 from 3,384 routine, composite, quarter milk samples of individual cows raised on 12 dairy farms in central Italy. Seventeen out of 60 isolates were confirmed as S. aureus after coagulase, thermonuclease, and biochemical tests. These isolates were analyzed by PCR for the presence of the nuc, sea, seb, sec, sed, and see genes. The positive isolates were nuc, 100% (17); sea, 35.29% (6); seb, 5.88% (1); sec, 5.88% (1); sed, 29.41% (5); and see, 47.06% (8). The isolates were also tested with 2 enzyme immunoassay diagnostic kits, one for the screening detection of the production of staphylococcal enterotoxins (SEA, SEB, SEC, SED, SEE) and one for the detection of specific enterotoxin produced by each isolate. Seven out of 17 (41.18%) were enterotoxin producers: 7 produced SEA (41.18%), 1 SEB (5.88%), 1 SEC (5.88%), 5 SED (29.41%), and 6 SEE (35.29%). To further characterize the isolates, they were analyzed by the Kirby Bauer test for susceptibility to 13 antimicrobials (ampicillin, ciprofloxacin, kanamycin, tetracycline, gentamicin, methicillin, nalidixic acid, erythromycin, amoxicillin/clavulanic acid, streptomycin, vancomycin, neomycin, and enrofloxacin), and we detected resistance to ampicillin (52.94%), nalidixic acid (70.59%), erythromycin (5.88%), and amoxicillin/clavulanic acid (17.65%). The isolates were sensitive to the main classes of antimicrobials used for the treatment of bovine subclinical mastitis. The presence of enterotoxin-producing isolates of S. aureus in bovine milk means that a temperature abuse or a breakdown in the thermal treatment of the milk could present a food safety risk, particularly if all enterotoxigenic isolates could potentially produce SEA in milk.     

Antimicrobial resistance and virulence of Enterococcus faecalis isolated from retail food

Although enterococci are considered opportunistic nosocomial pathogens, their contribution to foodborne illnesses via dissemination through retail food remains undefined. In this study, prevalence and association of antimicrobial resistance and virulence factors of 80 Enterococcus faecalis isolates from retail food items were investigated. The highest rates of resistance were observed for lincomycin (73 of 80 isolates, 91%) and bacitracin (57 of 80 isolates, 71%), and lower rates of resistance (< or =40%) were found for chloramphenicol, ciprofloxacin, erythromycin, flavomycin, gentamicin, kanamycin, nitrofurantoin, penicillin, and tylosin. Overall resistance to antimicrobials was low for most isolates tested. Of the virulence factors tested, the majority of isolates were positive for ccf (78 of 80 isolates, 98%), efaAfs (77 of 80, 96%), and cpd (74 of 80, 93%). Isolates also commonly contained cob (72 of 80 isolates, 90%) and gelE (68 of 80, 85%). Very few isolates contained cylMBA (12 of 80 isolates [15%] for cylM and 9 of 80 isolates [11%] for both cylB and cylA) and efaAfm (2 of 80 isolates, 3%). Positive statistical associations (significance level of 0.05) were found between agg and tetracycline resistance, cylM and erythromycin resistance, and gelE and efaAfs and lincomycin resistance. The presence of the cylB and cylA alleles also was positively correlated with bacitracin and tetracycline resistance. Negative correlations were observed between many of the virulence attributes and resistance to ciprofloxacin, erythromycin, flavomycin, gentamicin, kanamycin, and tylosin. These data suggest that both positive and negative associations exist between antimicrobial resistance genes and virulence factors in E. faecalis isolates from foods commonly purchased from grocery stores.     

Short communication: Association between the accessory gene regulator (agr) group and the severity of bovine mastitis caused by Staphylococcus aureus

Staphylococcus aureus can elicit mild to more severe degrees of mastitis in cattle, depending on the response of the host's immune system and the virulence factors of the specific isolate. Several virulence factors are controlled by a global regulatory system, designated accessory gene regulator (agr). Thus, the objective was to examine associations between different capsular and agr types and the severity of bovine mastitis caused by S. aureus. All isolates were obtained from bovine subclinical (n = 50), mild clinical (n = 73), and moderate clinical mastitis cases (n = 28). Isolates containing the agrI gene and lacking the agr locus (agr^?) were more prevalent among subclinical than clinical mastitis cases, whereas isolates containing the agrII and agrIII genes were more prevalent among clinical mastitis cases. The capsular types 5 (cap5) and 8 (cap8) were found in 42 and 44%, respectively, of the isolates obtained from subclinical cases and in 38.6 and 58.4%, respectively, of those isolated from clinical mastitis cases. Capsular type was not associated with type of mastitis (subclinical, mild clinical, or moderate clinical). We found a strong association between agr type and type of mastitis, suggesting that knowledge of S. aureus genetic profiles could be an additional tool to control this disease.     

Prevalence,characterization and antibiotic resistance of enterococci from traditional cheeses in Turkey

The aim of the present work is to provide information about Enterococcus strains isolated from traditional Turkish cheese samples in Ankara (Turkey), focusing on their prevalence, phenotypic and genotypic characteristics, and antibiotic resistance. A total of 213 probable enterococcal isolates isolated from 215 samples were identified by phenotypic and genotypic methods. As a result of 16S rDNA sequence analysis, 88 of the 213 enterococci strains were identified as Enterococcus faecium and 125 as Enterococcus faecalis. The E. faecalis strains (58.7%) were identified as the dominant species isolated from cheese samples in Turkey. The 213 Enterococcus strains were tested for susceptibility to 12 different antimicrobial agents. The resistance phenotype were as follow: nalidixic acid (100%), kanamycin (98.6%), rifampicin (78.4%), ampicillin (48.8%), ciprofloxacin (45.5%), erythromycin (18.8%), tetracycline (11.7%), penicillin G (5.6%), chloramphenicol (4.2%), gentamycin (3.8%) and streptomycin (1.4%). None of the strains was resistant to vancomycin. E. faecium strains showed more resistant phenotypes than E. faecalis strains as shown by the antibiotic resistance levels. It was also observed that the resistance of E. faecium and E. faecalis strains against the antibiotics was statistically significant (p ? 0.05). In total, 100% of E. faecium and 88.8% of E. faecalis strains were resistant to multiple drugs.     

Enterococcus populations in artisanal Manchego cheese: biodiversity, technological and safety aspects

Enterococci represent a considerable proportion of the microbiota in Manchego cheeses. In this study, a total of 132 enterococci isolated from good quality Manchego cheeses from two dairies at different ripening times were genotypically characterized and identified using molecular techniques. Representative isolates from the clusters obtained after genotyping were assayed for some enzymatic activities considered to have a potential role in cheese ripening, and for 2,3-butanedione and acetoin production, evaluation of odor intensity and appearance in milk and safety evaluation. Enterococcus faecalis was the predominant specie, accounting for 81.8% of the total isolates, while Enterococcus faecium, Enterococcus hirae and Enterococcus avium were present in low proportions. The number of genotypes involved at each ripening time varied both between dairies and with the ripening times; genotype E. faecalis Q1 being present in almost all the samples from both dairies. Eight isolates showed a higher proteolytic activity and 3 isolates produced high quantities of acetoin-diacetyl, for which reason they are interesting from a technological standpoint. A low antibiotic resistance was found and almost all the strains were susceptible to clinically important antibiotics. On the contrary, only four isolates (E. faecalis C4W1 and N0W5, and E. faecium N32W1 and C16W2) did not harbor some of the virulence genes assayed.     

Identification and antimicrobial resistance of Streptococcus uberis and Streptococcus parauberis isolated from bovine milk samples

The conventional identification of Streptococcus uberis/parauberis group (n = 137) in clinical and subclinical bovine mastitis samples originating from 111 different farms was compared with identification based on 16 and 23S rRNA gene HindIII RFLP patterns used as operational taxonomic units in numerical analyses. On the basis of ribopattern analysis only 2 isolates belonged to S. parauberis, which is thus not a frequent cause of bovine intramammary infections in Finland. According to in vitro antimicrobial susceptibility testing, Streptococcus uberis is susceptible to β-lactam antibiotics. The prevalence of erythromycin (15.6%) and oxytetracycline (40.6%) resistance of clinical S. uberis isolates was higher than reported previously among subclinical isolates. The 2 subclinical S. parauberis isolates were susceptible to all the antimicrobials tested.     

自贡市肉及其制品中粪肠球菌耐药性毒力基因和多位点序列分析

目的了解四川省自贡市肉及其制品分离粪肠球菌的抗生素耐药性和携带毒力基因情况,以及多重耐药菌株的序列分型(ST)。方法 2013年4月17~21日对147份肉及其制品样品污染的粪肠球菌进行分离鉴定,使用全自动微生物鉴定仪对分离菌株的耐药性进行检验,应用聚合酶链式反应(PCR)方法检测分离到的粪肠球菌携带4种常见毒力基因的情况,并对多重耐药菌株进行多位点序列分型(MLST)研究。结果从147份肉及其制品样品中共分离到65株粪肠球菌,其中耐药菌株比例为58.5%(38/65);分离菌株对四环素的耐药率最高,为41.5%(27/65),对利福平、氯霉素和红霉素也均有较高的耐药率;分离菌株对高浓度链霉素和高浓度庆大霉素的耐药率分别达到了15.4%(10/65)和12.3%(8/65);未分离到对青霉素类(青霉素和氨苄西林)、糖肽类(万古霉素)和脂肽类(达托霉素)抗生素耐药的菌株。4种常见毒力基因(gel E、asa1、esp、cyl A)在65株分离株中均有携带,阳性率分别为56.9%(37/65)、21.5%(14/65)、9.2%(6/65)和7.7%(5/65)。14株多重耐药菌株共有9个MLST型别,包括4株ST16、2株ST81和2株ST480菌株,且相同ST型别的粪肠球菌有相似的耐药谱和毒力基因携带情况。结论四川省自贡市肉及其制品中相同ST型别的粪肠球菌有相似的耐药谱和毒力基因携带情况,应重视肉及其制品中耐药粪肠球菌对公众健康的潜在威胁。