Extra-virgin olive oils contain many bioactive substances that are phenolic compounds. The survival of Arcobacter-like strains in non-buffered (WEOO) and buffered (BEOO) extracts of olive oils were studied. Time kill curves of different strains were measured in the environment of olive oil extracts of different grades. The activity of the extracts was also monitored for biofilm formation using the Christensen method. In vitro results revealed that extra-virgin olive oil extracts exhibited the strongest antimicrobial effects, especially non-buffered extracts, which exhibited strain inhibition after only 5 min of exposure. The weakest inhibitory effects were observed for olive oil extracts. A decrease in biofilm formation was observed in the environment of higher WEOO concentrations, although at lower concentrations of extracts, increased biofilm formation occurred due to stress conditions. The dialdehydic forms of oleuropein derivatives, hydroxytyrosol, and tyrosol were the main compounds detected by HPLC-CoulArray. The results indicate that not all olive oils had a similar bactericidal effect, and that bioactivity primarily depended on the content of certain phenolic compounds.
- Klíčová slova
- Arcobacter-like, HPLC, antimicrobial effect, biofilm formation, olive oil extract,
- MeSH
- Arcobacter * MeSH
- biofilmy MeSH
- fenoly analýza farmakologie MeSH
- oleje rostlin farmakologie MeSH
- olivový olej analýza MeSH
- plankton MeSH
- voda MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fenoly MeSH
- oleje rostlin MeSH
- olivový olej MeSH
- voda MeSH
Hydrolates obtained via the hydrodistillation and steam distillation of Lavandulaangustifolia Mill., Syzygiumaromaticum L., Foeniculumvulgare Mill., and Laurusnobilis L. were analyzed by gas chromatography with flame ionization detector (GC-FID) and gas chromatography coupled to mass spectrometry (GC-MS). Additionally, the hydrolates were evaluated for antimicrobial activity (disk-diffusion and microdilution method), influence on biofilm formation (Christensen method) and cytotoxicity of concentrated hydrolates against human cell lines (A549) by xCELLigence system. Using chemical analysis, 48, 9, 13 and 33 different components were detected in lavender, clove, fennel and laurel hydrolates, respectively. Lavender hydrolate contained the largest proportion of 1,8-cineol, linalool furanoxide, and linalool. The main components of laurel hydrolate were 1,8-cineol, 4-terpineol and α-terpineol. Fenchone and estragole were the most abundant in fennel hydrolate, and eugenol and eugenyl acetate in clove hydrolate. Concentrated hydrolates showed significant antimicrobial activity. Clove hydrolate was among the most antimicrobially active agents, most preferably against C. albicans, with an inhibition zone up to 23.5 mm. Moreover, concentrated hydrolates did not show any cytotoxic effect again8 st human A549 cells. In the presence of the non-concentrated hydrolates, significantly reduced biofilm formation was observed; however, with concentrated clove hydrolate, there was an increase in biofilm formation, e.g., of A. thereius, A. lanthieri, and A. butzleri. Research shows new findings about hydrolates that may be important in natural medicine or for preservation purposes.
- Klíčová slova
- Arcobacter-like bacteria, Foeniculum vulgare, Laurus nobilis, Lavandula angustifolia, Syzygium aromaticum, antimicrobial activity, biofilm formation, distillation, gas chromatography, hydrolates,
- MeSH
- antiinfekční látky farmakologie MeSH
- Arcobacter účinky léků MeSH
- buňky A549 MeSH
- Candida albicans účinky léků MeSH
- destilace MeSH
- levandule chemie MeSH
- lidé MeSH
- nádory plic farmakoterapie MeSH
- oleje prchavé farmakologie MeSH
- oleje rostlin farmakologie MeSH
- proliferace buněk MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antiinfekční látky MeSH
- oleje prchavé MeSH
- oleje rostlin MeSH
Arcobacter butzleri is an emerging human and animal pathogen for which an increased prevalence of resistance to antibiotics has been observed, and so alternative compounds to modulate resistance of A. butzleri are required. This work aims to study the potential use of several polyphenols as efflux pump inhibitors (EPIs) and to evaluate their interaction with antibiotics, in order to enhance antibiotic activity against A. butzleri. The minimum inhibitory concentration (MIC) of (-)-epicatechin, (+)-catechin, rutin, gallic acid, caffeic acid, chlorogenic acid, resveratrol, pterostilbene, and pinosylvin was determined, in absence and presence of four known EPIs. Subsequently, ethidium bromide accumulation in presence of subinhibitory concentrations of polyphenols was evaluated, and the synergistic potential of the compounds with antibiotics was assessed by checkerboard dilution test. Only stilbenes presented activity against A. butzleri, with MIC values ranging between 64 and 512 μg/mL. The MIC determination of the polyphenols in the presence of subinhibitory concentrations of known EPIs showed that efflux pumps play a role in the resistance to these compounds. Stilbenes also induced a higher intracellular accumulation of ethidium bromide, indicating that they may inhibit the activity of efflux pumps. Checkerboard assays showed that several combinations of polyphenol/antibiotic had an additive effect against A. butzleri. Overall, the results indicate that some polyphenols reduce A. butzleri resistance to antibiotics, suggesting the potential of stilbenes as EPIs. The potential of resveratrol and pinosylvin as resistance modulators was evidenced, insofar as these compounds can even revert antibiotic resistance. Therefore, the use of polyphenols as resistance modulators could be an alternative to overcome the decreasing susceptibility of A. butzleri to antibiotics.
- MeSH
- antibakteriální látky farmakologie MeSH
- Arcobacter účinky léků genetika metabolismus MeSH
- bakteriální léková rezistence MeSH
- bakteriální proteiny antagonisté a inhibitory genetika metabolismus MeSH
- gramnegativní bakteriální infekce mikrobiologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- polyfenoly farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- bakteriální proteiny MeSH
- polyfenoly MeSH
Bacteria of the Arcobacter (A.) genus, originating mainly from food and water, are dreaded germs for humans as well as animals. However, the virulence of these bacteria has not been fully elucidated yet. This study looked at the occurrence of eight virulence-associated factors (ciaB, cj1349, pldA, irgA, hecA, tlyA, mviN, hecB) in a total of 80 isolates of Arcobacter butzleri and 22 isolates of A. cryaerophilus. The isolates were derived from food, water, and clinical samples. A polymerase chain reaction using specific primers was used to detect these virulence-associated genes. The presence of all genes in the isolates of A. butzleri (98.8% ciaB, 95.0% cj1349, 98.8% pldA, 22.5% irgA, 31.3% hecA, 95.0% tlyA, 97.5% mviN, 38.8% hecB) and A. cryaerophilus (95.5% ciaB, 0.0% cj1349, 9.1% pldA, 0.0% irgA, 0.0% hecA, 31.8% tlyA, 90.9% mviN, 0.0% hecB) was monitored. Among the tested isolates, there were 13 isolates (12.7%) of A. butzleri, in which the presence of all eight virulence-associated genes was recorded in the genome. In contrast, in one A. cryaerophilus strain, none of the observed genes were detected. The presence of ciaB and mviN genes was significantly more frequent in A. cryaerophilus isolates than other genes (P < 0.05). In general, more virulence-associated genes have been detected in A. butzleri isolates compared to A. cryaerophilus. The most common gene combination (ciaB, cj1349, pldA, tlyA, mviN) was detected in case of 39 isolates. In 50.0% of A. butzleri isolates derived from clinical samples, all eight virulence-associated genes were significantly more frequently detected (P < 0.05). The tlyA gene occurred significantly more frequent in A. butzleri isolates from meat and water samples and irgA and hecB genes in clinical samples. Therefore, our study provides information about occurrence of virulence-associated genes in genome of Arcobacter isolates. These findings could be hazardous to human health, because the presence of virulence-associated genes is the assumption for potential dangerousness of these bacteria. Our results indicate high incidence of virulence-associated genes in Arcobacter genomes and hence potentially pathogenic properties of the studied strains.
- MeSH
- Arcobacter genetika izolace a purifikace patogenita MeSH
- bakteriální geny genetika MeSH
- faktory virulence genetika MeSH
- genetická variace MeSH
- gramnegativní bakteriální infekce mikrobiologie MeSH
- mikrobiologie vody * MeSH
- potravinářská mikrobiologie * MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- faktory virulence MeSH
OBJECTIVES: Arcobacter spp. are considered to be potential foodborne pathogens, and consumption of contaminated food containing these bacteria could endanger human and animal health. Arcobacter butzleri and Arcobacter cryaerophilus are the species most frequently isolated from food of animal origin and from other samples. The aim of this study was to evaluate the susceptibility of arcobacters isolated in the Czech Republic. No information about antibiotic susceptibility and multidrug resistance of arcobacters isolated in the Czech Republic is available in the literature before now. METHODS: The antimicrobial resistance of A. butzleri (n=80) and A. cryaerophilus (n=20) isolated from meat of animal origin, water sources and clinical samples was examined by the disk diffusion method. RESULTS: Arcobacters were resistant to one or more antimicrobial agents in 99% (99/100) of tested isolates. Most of the Arcobacter isolates were resistant to β-lactam antibiotics, i.e. ampicillin (81.0%), amoxicillin/clavulanic acid (28.0%), cefalotin (73.0%) and aztreonam (93.0%). Arcobacters were also frequently resistant to lincosamides, i.e. clindamycin (98.0%). Of the aminoglycosides, amikacin, gentamicin and tobramycin were evaluated to be the most effective antibiotics among those tested against arcobacters. CONCLUSIONS: These results demonstrate substantial resistance in Arcobacter isolates to 18 antimicrobial agents commonly used in medical and veterinary medicine. Multidrug resistance was found in 93.8% (75/80) of A. butzleri isolates and 70.0% (14/20) of A. cryaerophilus isolates.
- Klíčová slova
- Antibiogram, Antibiotic, Antimicrobial susceptibility testing, Arcobacter spp., Susceptibility,
- MeSH
- antibakteriální látky farmakologie MeSH
- Arcobacter účinky léků izolace a purifikace MeSH
- diskové difúzní antimikrobiální testy MeSH
- gramnegativní bakteriální infekce mikrobiologie MeSH
- lidé MeSH
- mikrobiologie vody * MeSH
- mnohočetná bakteriální léková rezistence * MeSH
- potravinářská mikrobiologie * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- antibakteriální látky MeSH
Water plays an important role in the transmission of Arcobacter spp. to animals and humans. The aim of this study was to isolate and characterize Arcobacter spp. from 115 different water samples (66 sewage, 25 rivers, 16 spring water, and 8 drinking water) in Izmir, Turkey. In total, 41 samples (35.7 %) were found positive for Arcobacter spp. by the genus-specific PCR. Arcobacter butzleri was detected in 39 out of 115 samples (33.9 %) including 24 sewage, 13 rivers, and 2 spring water. The remaining Arcobacter spp. (n = 2) isolates could not be identified by m-PCR and 16S rRNA gene sequencing. Based on the phenotypic characterization, most of the Arcobacter species (87.8 %) indicated weak catalase activity. In addition, there were differences in phenotypic patterns among isolated species during growth at 37 °C under microaerobic and aerobic conditions, in the presence of 2 % (39/41) and 3.5 % (32/41) NaCl and 0.04 % TTC (39/41) and on MacConkey agar (38/41). The results of this study indicated that environmental water samples are common sources for Arcobacter spp. Therefore, effective control measures should be taken to protect human health.
- MeSH
- aerobióza MeSH
- Arcobacter klasifikace genetika růst a vývoj izolace a purifikace MeSH
- chlorid sodný metabolismus MeSH
- DNA bakterií chemie genetika MeSH
- kultivační média chemie MeSH
- mikrobiologie vody * MeSH
- ribozomální DNA chemie genetika MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- techniky typizace bakterií MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Turecko MeSH
- Názvy látek
- chlorid sodný MeSH
- DNA bakterií MeSH
- kultivační média MeSH
- ribozomální DNA MeSH
- RNA ribozomální 16S MeSH
This study provides information on the occurrence of Arcobacter in several types of water and food products of animal origin in the Czech Republic. We processed 190 samples using the modified method, and the occurrence of Arcobacter spp. was confirmed in 36.8 % of these. This total incidence consisted of Arcobacter butzleri (27.3 %), Arcobacter cryaerophilus (8.4 %) and Arcobacter skirrowii (1.1 %). We newly described the common presence of Arcobacter spp. in sewage water in the Czech Republic that is released into waterways after processing in water treatment plants (86.7 %). All the acquired isolates were subject to detailed confirmation with subsequent species classification using multiplex PCR and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In this study, we used a modification of a method using passive filtration of an enriched sample, which could be suitable for the isolation of Arcobacter, especially in combination with Campylobacter selective agar chromogenic medium. Our studies have shown this agar to be quite suited to the isolation of Arcobacter and that it can be an appropriate instrument for accelerating culture diagnostics.
- MeSH
- Arcobacter genetika růst a vývoj izolace a purifikace metabolismus MeSH
- kultivační média metabolismus MeSH
- odpadní vody mikrobiologie MeSH
- počet mikrobiálních kolonií přístrojové vybavení metody MeSH
- potravinářská mikrobiologie * MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- kultivační média MeSH
- odpadní vody MeSH
The genus Arcobacter is related to the well-known human pathogen, Campylobacter jejuni, and has been linked to human diseases. In this study, the survival of Arcobacter spp. in various concentrations of ethanol, in various samples of beers, and in a model stomach has been investigated. For most of these bacteria, a concentration of 10 % ethanol was determined to be the minimum inhibitory concentration. The fact that these organisms are able to survive under these conditions may have an impact in the food processing industry. We studied the activity of beer against arcobacters. These bacteria were killed in all samples of beer within 30 min. A model stomach, containing a food matrix and a synthetic gastric fluid, was used to deduce the effect of beer against Arcobacter spp. during food consumption. Complete inactivation of all monitored arcobacters was detected usually within 15 min. However, the presence of beer does not potentiate the effect of gastric fluid against these bacteria. This is apparently the first study focusing upon the effect of beer on Arcobacter spp.
- MeSH
- Arcobacter účinky léků fyziologie MeSH
- časové faktory MeSH
- ethanol toxicita MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- mikrobiální viabilita účinky léků MeSH
- pivo analýza mikrobiologie MeSH
- teoretické modely MeSH
- žaludek mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ethanol MeSH
The inhibitory effects of 17 organic acids (C₂-C₁₆ fatty acids, sorbic, benzoic, phenylacetic, fumaric, succinic, lactic, malic and citric) on Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii were investigated by determining their IC₅₀ values, defined as the concentration of acid at which the target DNA sequence was expressed at 50% of the positive control level in cultures incubated at 30°C for 24 h. DNA was analysed by real-time PCR. The Arcobacter strains tested were inhibited by all the organic acids, with the sensitivities in the order A. skirrowii > A. cryaerophilus > A. butzleri. Eight acids with IC₅₀ values of <1 mg/mL against A. butzleri were tested for their effects on A. butzleri inoculated on chicken carcasses at a concentration of 5 log CFU/g of skin. Inoculated halved carcasses were immersed in solutions of the acids at 5 mg/mL for 1 min. Samples of skin were collected from carcass halves after storage at 4°C for 0, 1, 2 or 3 days for enumeration of arcobacters on Muller-Hinton agar. All eight tested acids suppressed bacterial proliferation. The highest inhibitory activities were observed for benzoic, citric, malic and sorbic acids. Subsequent sensory analysis revealed benzoic acid to be the most suitable organic acid for chicken skin treatment.
- MeSH
- Arcobacter účinky léků růst a vývoj MeSH
- inhibiční koncentrace 50 MeSH
- kur domácí MeSH
- kůže mikrobiologie MeSH
- kyseliny farmakologie MeSH
- manipulace s potravinami MeSH
- maso mikrobiologie normy MeSH
- organické látky farmakologie MeSH
- potravinářská mikrobiologie * metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kyseliny MeSH
- organické látky MeSH
The aim of this study was to examine 634 samples of chicken, lamb, pork, beef, fish, samples from the intensive animal industry and from poultry for slaughter, as well as from the domestic breeding of poultry, horses, pigs, and lambs, from surface water, and from clinical samples for the presence of Arcobacter. All the samples were examined with a cultivation method, followed by confirmation by multiplex PCR. The method of multiplex PCR applied directly to a liquid medium after enrichment was applied only to the samples with the highest probability of the presence of arcobacters. Arcobacter spp. were detected in 11.8% of the samples, of which A. butzleri, A. cryaerophilus, and A. skirrowii were found in 6.6, 5.1, and 0.2% of the samples, respectively. The sources of the arcobacters were chicken meat from the retail market, intensive animal production facilities, domestic chicken breeding facilities, lamb raising environments, surface water and wastewater, and beef swabs taken in a meat processing factory. No occurrence of arcobacters was identified in the swabs from slaughter turkeys, ducks, and wild poultry. No arcobacters were found in horse and pig breeding environments, on pork, or on the swabs of fish. Forty-two rectal swabs taken from humans were also free of Arcobacter. Seventeen isolates of Arcobacter were further identified by sequencing the 16S rRNA gene. Varied genotypes were observed among A. butzleri from chicken meat and chicken breeds, and A. cryaerophilus from wastewater and chicken breeds. They were similar to the genotypes present in wastewater, porcine feces, human stool, and human blood obtained from databases. Our results revealed that the chicken meat from the retail market is an important source of arcobacters. Cross-contamination during handling of chicken carcass practices could play a key role in the spread of Arcobacter.
- MeSH
- Arcobacter klasifikace izolace a purifikace MeSH
- druhová specificita MeSH
- kontaminace potravin analýza MeSH
- lidé MeSH
- manipulace s potravinami metody MeSH
- maso mikrobiologie MeSH
- počet mikrobiálních kolonií MeSH
- polymerázová řetězová reakce metody MeSH
- potrava z moře (živočišná) mikrobiologie MeSH
- potravinářská mikrobiologie MeSH
- prevalence MeSH
- spotřebitelská bezpečnost produktů MeSH
- techniky typizace bakterií metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
