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The objective of this study was to examine antibacterial, antifungal and antiviral properties of selected Phlomis species (Lamiaceae) growing in Turkey. The petroleum ether and methanol extracts of the seven species, namely P. armeniaca Willd., P. bourgaei Boiss., P. leucophracta P.H. Davis & Hub.-Mor., P. lunariifolia Sibth. & Sm., P. lycia D. Don, P. pungens Willd. var. pungens , and P. pungens var. hirta Velen. were tested against Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Bacillus subtilis , and Enterococcus faecalis for their antibacterial activity using ampicillin and oflaxocin as references. Antifungal activity of the same extracts was determined against Candida albicans using microdilution method with ketocanazole as reference. Both DNA virus Herpes simplex type-1 (HSV-1) and RNA virus Parainfluenza (PI-3) were employed for antiviral assessment of the Phlomis extracts using Madin-Darby Bovine Kidney and Vero cell lines in which acyclovir for HSV-1 and oseltamivir for PI-3 were employed as reference drugs. Although both the petroleum ether and methanol extracts seemed to exert similar antibacterial activity, the methanolic extracts were observed to be more active against S. aureus and E. faecalis . On the other hand, methanolic extract of P. pungens var. pungens possessed notable antiviral activity against both type of viruses.
Plasmid content was investigated in hundred copiotrophic Gram-negative river water isolates that exhibited resistance to four or more antibiotics. A total of seventy-seven isolates were found to carry plasmids of varying sizes. These isolates were primarily grouped as Pseudomonads and members of Enterobacteriaceae on the basis of physiological and biochemical tests. Fifty-six isolates that were rifampicin-sensitive and belonged to Enterobacteriaceae family were chosen as donors for the conjugal transfer assay. Eighteen of the isolates successfully transferred conjugable plasmids to the E. coli DH5 α recipient. Countable multiple antibiotic resistant transconjugants arose readily and conjugal transfer frequency was in the range of 3.75 × 10 −6 to 1.0 × 10 −1 . The most common carriage of resistances conferred by transmissible R plasmids was against ampicillin, cefotaxim and cephalexin. The residence of class 1 integrons on conjugative R plasmids was confirmed in only six transconjugants. Gene cassettes borne on the integrons were identified to be dihydrofolate reductases (dhfrs) . The major concern of this study was about the copiotrophs containing self-transmissible R plasmids which may be potential reservoirs of antibiotic-resistance genes and instrumental in dissemination of the same in the environment.
This study was conducted at a 900+ bed general teaching hospital, from May to September 2007, in Iran. The aim of this study was to determine the prevalence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae and their antimicrobial pattern. The Kirby-Bauer disk diffusion method and the phenotypic disk confirmatory test were performed for each isolate. The total of 206 isolates including 106 E. coli and 100 K. pneumoniae were collected of which 122 isolates (59.2%) were ESBL positive. The prevalence of ESBL-producing strains was 59.2% (122/206). All the isolates were susceptible to imipenem. Among the ESBL-producing isolates, the sensitivity was from 3.3% to 61.5% for ampicillin to aztreonam. From female isolates (136), 59.5% and from male isolates (70), 58.6% were ESBL-producers. Ratios of isolates from hospitalized patients to out-patients were 94/28 in the ESBL-producing group. The number of ESBL-producing isolates according to the isolation sites showed a significant difference between ESBL-producers and non-producers in blood samples ( P <0.05). This study shows that the prevalence of ESBL strains in Iran is high. It seems necessary for clinicians and medical community personnel to be fully aware of ESBL-producing microorganisms.
L. monocytogenes poses a serious threat to public health, since most cases of listeriosis are connected with eating contaminated food. L. monocytogenes is often detected both in fresh and frozen vegetables.
The aim of this study was to evaluate the antibiotic susceptibility and ability to form biofilm of L. monocytogenes strains isolated from frozen vegetable mixtures in Poland.
Ninetynine genetically different strains were found among 100 isolates of L. monocytogenes. Among the 99 strains, 80 (80.8%) were susceptible to all tested antibiotics. Nineteen (19.2%) strains were resistant to one or more antibiotics. From this group of L. monocytogenes strains, most strains were resistant to erythromycin (16; 16,1%), penicillin (15; 15.1%), meropenem (12; 12.1%), cotrimoxazole (12; 12.1%), and ampicillin (3; 3.1%). According to the obtained results, differences in intensity of biofilm, both between those isolated in successive years and in the particular year, were observed. Performed analysis showed statistically insignificant faint negative correlation (r=–0.088) between the number of antibiotics to which strains were resistant and the intensity of biofilm formation by them.
Food contamination with L. monocytogenes poses a threat to consumers, therefore it is necessary to monitor their antibiotic susceptibility, ability to form biofilm, and genetic similarity, in order to evaluate the strains persistence time in plant.
Honey is the most important bee product. There are many secondary metabolites, carbohydrates, enzymes, and vitamins in honey, thus, honey has antimicrobial activity. In this study, in vitro antimicrobial activity of forty-two honey and eight propolis ethanolic extracts (PEE) were investigated against 16 microorganisms. Total phenolic content ranged between 20.00–124.10 mg GAE/100 g and 103–232 mg GAE/g for honey and raw propolis samples, respectively. Pine and oak honeydew honeys had higher antimicrobial activity than four different grades of Manuka Honeys up to 18 mm minimum inhibition zone diameters. The ethanolic propolis extracts showed much higher antimicrobial activity than the honey samples. Fungi species were inhibited by the propolis samples. Helicobacter pylorii (H. pylorii) was the most sensitive, whereas Streptococcus agalactiae was the most resistant bacteria among the studied microorganisms. Brazilian and Zonguldak propolis had the closest antimicrobial activity to ampicillin, streptomycin, and fluconazole. It can be concluded that both honey and propolis could be used in preservative and complementary medicine.
Excessive use of antibiotics leads to their occurrence into the environment. In spite of their benefit properties and desired effects during the therapeutic applications, the same properties can be disadvantageous for the environment having negative influences over the plants and microorganisms and the potential risks for human health. Regarding the monitoring of antibiotics and their subsequent elimination from environment, it is necessary to develop analytical procedures for their determination. In the present study, the quantitative determination of seven antibiotics belonging to three different classes is reported: tetracyclines (tetracycline and doxycycline), cephalosporins (ceftazidime and ceftriaxone), and penicillins (amoxicillin, ampicillin, and penicillin G) from surface waters. The proposed procedure consists of the solid phase extraction (SPE) of studied antibiotics from river water samples, their separation by high-perfomance thin-layer chromatography (HPTLC), and quantification by UV densitometry. The antibiotics were extracted from water matrices using hydrophilic-lipophilic-balanced Oasis HLB cartridges. The cartridge efficiency of the SPE method was checked by recovery experiments and evaluated by HPTLC. The chromatographic separation was performed on pre-coated Alugram SIL G/UV254 plates with ethyl acetate-methanol-acetone-water 5:2.5: 2.5:1.5 (v/v) mobile phase. The bands were detected and quantified at 254 nm by densitometry. For method validation, studies of selectivity, linearity, limits of detection and quantification, and precision and accuracy were achieved. The proposed procedure was applied to the determination of studied antibiotics on surface water samples collected from Someoul Mic River (Romania).
The aims of this study were to isolate LAB from Thai plant-derived foods and beverages and to examine in vitro probiotic properties including b-glucosidase enzyme activity. Lactobacillus plantarum SC 359 selected from Thai pickled soybean was significantly (P<0.05) the best to perform β-glucosidase enzyme activity (0.396 U ml−1 at 18 h of incubation) out of the 227 tested strains. The strain survived in 0.30% (w/v) bile salt and had high tolerance to acidic pH with survival rates at a 2 h period of 72.24%, 85.52%, 92.64%, and 93.38% at pH 2.0, 3.0, 4.0, and 5.0, respectively. The SC 359 strain showed proteolytic and lipolytic activities. Moreover, the selected strain displayed strong antagonistic activities against Bacillus cereus ATCC 11778, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Salmonella Typhi, Shigella sonnei and Candida albicans ATCC 90028. The strain was susceptible to ampicillin, chloramphenicol, erythromycin, gentamicin, kanamycin, rifampicin, streptomycin, tetracycline, and vancomycin. In addition, the selected strain significantly inhibited the adherence to Caco-2 cells of E. coli, S. Typhi and Sh. sonnei (P<0.05) by 33.50 to 73.37%. The strain could obstruct the adherences of pathogens by elimination, competition, and displacement with pathogen adherences 33.62–53.92%, 26.63–59.23%, and 49.41–66.50%, respectively. Based on the results, the selected strain could be applied as functional starter for Thai fermented plant-derived foods and beverages.
The biochemical characteristics and antibiotic susceptibility of 12 Ornithobacterium rhinotracheale strains isolated from chickens and turkeys suffering from respiratory clinical signs and the survival of some isolates on egg-shell and within chicken eggs during hatching were examined. All O. rhinotracheale strains showed typical biochemical characteristics. Among the 16 drugs examined, penicillin G, ampicillin (MICs ranging from ≤ 0.06 μg/ml to 1 μg/ml), ceftazidim (with MICs from ≤ 0.06 μg/ml to 0.12 μg/ml), erythromycin, tylosin, tilmicosin (with some exceptions MICs ranged from ≤ 0.06 μg/ml to 1 μg/ml) and tiamulin (MICs varied from ≤ 0.06 μg/ml to 2 μg/ml) were the most effective. Lincomycin, oxytetracycline and enrofloxacin also gave good inhibitions, but with most strains in a higher concentration (MICs ranged in most cases from 2 μg/ml to 8 μg/ml). The other antibiotics inhibited the growth of O. rhinotracheale only in very high concentrations (colistin) or not at all (apramycin, spectinomycin, polymyxin B). At 37 °C, O. rhinotracheale did not survive on egg-shell for more than 24 hours, while upon inoculation into embryonated chicken eggs it killed embryos by the ninth day, and from the 14th day post-inoculation no O. rhinotracheale could be cultured from the eggs at all. These results suggest that O. rhinotracheale is not transmitted via eggs during hatching.
Besides the well-known O157:H7 clone causing enterohaemorrhagic colitis and haemolytic uraemic syndrome in Europe, Japan and North America, the number of Escherichia coli isolates with non-motile (NM) phenotype has considerably increased. We supposed that spontaneous antibiotic resistance mutation could cause this phenotypic change. To model our hypothesis we isolated rifampicin- (Rif) and ampicillin- (Amp) resistant mutants from E. coli O157:H7 prototype strains 7785 and EDL933. Among Rif r mutants we could isolate strains with no or reduced motility, while the Ampr mutants became hypermotile. The biochemical profile of the mutants had not changed but phage sensitivity and generation time of the mutants were altered. Among the representative strains we did not find polymorphism with Southern blot analysis and no polymorphism was found in the fliC gene of the mutants. The described characteristics have proven to be stable. In a mice virulence assay by intravenous infections the virulence of the derivatives was also found to be changed. In summary, we found that the antibiotic-resistant phenotype in E. coli O157:H7 was coexpressed with several other phenotypic changes including motility and virulence. It can be assumed that expression of the involved phenotypes may be under the influence of a common regulatory cascade. Further work is needed to identify the components and mechanism of this regulatory system.
The aim of this work was investigation of clinically important amino acid substitutions of NDM-1 variants. A bla NDM-1 gene was cloned into expression vector pET100/D-TOPO. The sequence of NDM-1 variants with substituted amino acids was determined by ClustalW program. A pET100/D-TOPO + bla NDM-1 was used to generate the alanine mutations at different positions, such as NDM-2 (P28A), NDM-3 (D95A), NDM-4 (M154A), NDM-5 (V88A), NDM-7 (D130A), and NDM-9 (E152A). The mutant variants were transformed into Escherichia coli DH5α. Changes in the activities of alanine mutation variants were determined by E-test. All samples had 32 μg/ml MIC values against ampicillin. The 28th amino acid mutation sample had the highest MIC value against ceftazidime, whereas decreased MIC value for piperacillin. It was observed that the resistance to imipenem was increased in mutant variants D95A, M154A, D130A, and E152A, comparing with P28A and V88A. It was found that NDM-1 has 0.64 μg/ml and the 130th amino acid mutation sample has 0.75 μg/ml meropenem MIC value.
