Nefise Akçelik

Biofilm-producing abilities of Salmonella strains isolated from Turkey

In the present study the biofilm-forming characteristics of 99 serotyped (DMC strains) and 41 genus level-identified (IS strains) Salmonella strains originating from Turkey were investigated. The strains were selected based on their ability to show the biofilm morphotype on Congo red agar plates. In addition, all strains were evaluated with regard to properties related to forming pellicle structures, physical differences of pellicles, any changes in the media associated with the formation of pellicles, and the presence of cellulose within the formed biofilm matrix as determined using 366 nm UV light. The Salmonella Typhimurium DMC4 strain was the best producer of biofilm grown on polystyrene microtiter plates (optical density at 595 nm: 3.418). In subsequent experiments industrial process conditions were used to investigate different morphotyped Salmonella strains’ biofilm-forming capability on stainless steel, a commonly preferred surface for the food industries, and on polystyrene surfaces. The effect of other important industrial conditions, such as temperature (5, 20, 37°C), pH (4.5, 5.5, 6.5, 7.4) and NaCl concentration (0.5, 1.5, 5.5, 10.5%) on the production of biofilm of the different morphotyped Salmonella strains (DMC4; red, dry and rough morphotyped S. Typhimurium, DMC12; brown, dry and rough morphotyped S. Infantis, DMC13; pink, dry and rough morphotyped S. subsp. Roughform) were also assessed. On the other hand, pH values exhibited variable effects on biofilm-forming features for different Salmonella strains on both polystyrene and stainless steel surfaces.

The interactions between esp, fsr, gelE genes and biofilm formation and pfge analysis of clinical Enterococcus faecium strains

Enterococcus faecium has become an increasingly important nosocomial pathogen due to formation of biofilms on several surfaces. Sixty one (61) E. faecium strains isolated from blood, urine and fecal were assessed for biofilm production, the effect of different glucose concentration on biofilm production and also the presence of esp, fsr and gelE genes. Pulsed field gel electrophoresis (PFGE) method was performed to show chromosomal similarities and also to determine correlation between biofilm formation ability and genetic identity of E. faecium strains. It was observed that glucose concentration of the medium and incubation period can affect biofilm formation of the bacteria. When tested strains were incubated in a medium containing 1% glucose for 48 h, 66.66% of urine isolates, 60.71% fecal isolates and 25% of blood isolates produced strong biofilm structures. esp-positive strains (80% of the all isolates) were also identified as strong biofilm producers compared to esp-negative isolates. As a result of PFGE analyses, isolates numbered 14 (isolated from fecal sample) and 81 (isolated from blood sample) were classified in minor group B at a level of 48% similarity. Out of these two isolates, all the isolates were included in major group A with 43% similarity level and this group was subdivided into six subgroups.

Identification and characterisation of lactic acid bacteria isolated from traditional Urfa cheese

In this study, characterisation of dominant strains of lactic flora in traditional Urfa cheese made from sheeps milk was performed using biochemical, phenotypic and genotypic methods. According to the results obtained, the percentage distributions of the lactic acid bacteria isolated were as follows: 48.95% Enterococcus spp., 40.55% Lactococcus spp., 9.10% Lactobacillus spp., 0.69% Streptococcus spp. and 0.69% Leuconostoc spp. The majority of lactococcal isolates showed an atypical phenotype of growing in the presence of 6.5% NaCl. Acidification and bacteriocin production were also determined for each isolate. Four strains showed bacteriocin activity.

Incidence of antibiotic resistance and virulence determinants in Enterococcus faecium and Enterococcus faecalis strains, isolated from traditional cheeses in Turkey

Enterococcus faecalis (n = 15) and Enterococcus faecium (n = 33) strains isolated from traditional Turkish cheeses were tested for susceptibility to 11 different antimicrobial agents and for the presence of selected genes encoding resistance and 13 genes encoding virulence factors using PCR. Furthermore, the plasmid profile of enterococci was examined. All E. faecium and E. faecalis isolates were resistant to nalidixic acid and kanamycin. The percentages of other resistant E. faecium and E. faecalis isolates, respectively, were 97 and 100% to streptomycin, 15.1 and 20% to ampicillin, 9.1 and 26.7% to gentamycin, 12.1 and 46.6% to chloramphenicol, 12.1 and 60% to tetracycline, 75.7 and 93.3% to rifampicin, 84.8 and 80% to vancomycin, 97 and 100% to erytromycin and 72.7 and 60% to ciprofloxacin. efaAfm (100%) and ccf (90.1%) genes were the most common virulence genes identified among E. faecium isolates while efaAfs (100%), cpd (100%), ccf (93.3%) and cob (86.7%) genes among E. faecalis isolates. Cytolysin determinants (cylM, cylB, cylA) were not detected among tested strains. Plasmid profile analysis of Enterococcus spp. revealed plasmid DNA bands ranging in size from 2.4 to 35.8 kb.

Modeling and predicting the biofilm formation of Salmonella Virchow with respect to temperature and pH

Biofilm formation of Salmonella Virchow was monitored with respect to time at three different temperature (20, 25 and 27.5 °C) and pH (5.2, 5.9 and 6.6) values. As the temperature increased at a constant pH level, biofilm formation decreased while as the pH level increased at a constant temperature, biofilm formation increased. Modified Gompertz equation with high adjusted determination coefficient (Radj(2)) and low mean square error (MSE) values produced reasonable fits for the biofilm formation under all conditions. Parameters of the modified Gompertz equation could be described in terms of temperature and pH by use of a second order polynomial function. In general, as temperature increased maximum biofilm quantity, maximum biofilm formation rate and time of acceleration of biofilm formation decreased; whereas, as pH increased; maximum biofilm quantity, maximum biofilm formation rate and time of acceleration of biofilm formation increased. Two temperature (23 and 26 °C) and pH (5.3 and 6.3) values were used up to 24 h to predict the biofilm formation of S. Virchow. Although the predictions did not perfectly match with the data, reasonable estimates were obtained. In principle, modeling and predicting the biofilm formation of different microorganisms on different surfaces under various conditions could be possible.

Genetic diversity of food originated Salmonella isolates

ABSTRACT Currently, Salmonella enterica is the most common bacterial foodborne pathogen, causing serious extraintestinal disease. Typing methods play an important role on pathogens’ source tracking, knowing the source(s) of bacteria in pharmaceutical sciences, preventing and controlling the diarrhea and food-poisoning outbreaks. The purpose of this study is to use different moleculer typing methods to determine the genetic variability of 38 foodborne Salmonella isolates that were previously identified by biochemical tests. The methods were evaluated by four molecular techniques including 16S rRNA sequencing, PFGE, PCR-RFLP and invA-spvC PCR. 16S rRNA sequencing results showed that four of the 38 isolates were Escherichia coli, Proteus mirabilis and Citrobacter murliniae, and the others were Salmonella enterica. Thirty-eight strains were subtyped by XbaI-PFGE into 20 profiles with different clusters, while they were subtyped by 16S rRNA-RFLP into 9 profiles with a single cluster. Out of two Salmonella isolates, the invasion gene (invA) was detected in all other Salmonella isolates (94%) and the virulence gene (spvC) was detected in 11% of Salmonella isolates. Our results suggested that the PFGE subtyping is the prominent method for the evaluation and benchmarking of molecular subtyping.

Probiotic and Antioxidative Properties of L. Lactis LL27 Isolated from Milk

ABSTRACT The aim of this study was to determine the probiotic properties of Lactococcus lactis subsp. lactis LL27, namely its resistance to bile salt, pepsin, pancreatin, acid and antibiotics. Moreover, the ability of L. lactis to inhibit the adhesion of Escherichia coli ETEC and Salmonella Typhimurium SL1344 to Caco-2 cells were examined and also the hydrophobicity, iron-ion chelating ability, and determination of α, α-diphenyl-β-picrylhydrazyl (DPPH) radical-scavenging ability of the organism were investigated. L. lactis LL27, nisin A producer, was able to inhibit the growth of Gram-positive bacteria including spoilage and pathogenic strains. L. lactis LL27 exhibited high levels of resistance to low pH, bile salts, pepsin and pancreatin, which are conditions characteristic of the gastrointestinal tract environment. LL27 was found to adhere to Caco-2 and CCL-221 cell lines at a rate of 13 ± 2.3% and 29 ± 2.6%, respectively. In addition its presence reduced the ability of pathogenic bacterial species Escherichia coli ETEC and Salmonella Typhimurium SL 1344 to adhere to Caco-2 cells. The hydrophobicity, DPPH radical-scavenging activity and iron ion-chelating ability of the tested LL27 were found to be 83.48 ± 1.45%, 75 ± 3% and 29.5 ± 3%, respectively. Scanning electron microscopy revealed the LL27 to be located inside Caco-2 and CCL-221 mammalian cells. These findings suggested that L. lactis LL27 has good probiotic properties and could be used in functional foods.