Specific Antimicrobial Activities Revealed by Comparative Evaluation of Selected Gemmotherapy Extracts.

Nowadays, unprecedented health challenges are urging novel solutions to address antimicrobial resistance as multidrug-resistant strains of bacteria, yeasts and moulds are emerging. Such microorganisms can cause food and feed spoilage, food poisoning and even more severe diseases, resulting in human death. In order to overcome this phenomenon, it is essential to identify novel antimicrobials that are naturally occurring, biologically effective and increasingly safe for human use. The development of gemmotherapy extracts (GTEs) using plant parts such as buds and young shoots has emerged as a novel approach to treat/prevent human conditions due to their associated antidiabetic, anti-inflammatory and/or antimicrobial properties that all require careful evaluations. Seven GTEs obtained from plant species like the olive (Olea europaea L.), almond (Prunus amygdalus L.), black mulberry (Morus nigra L.), walnut (Juglans regia L.), blackberry (Rubus fruticosus L.), blackcurrant (Ribes nigrum L.) and bilberry (Vaccinium myrtillus L.) were tested for their antimicrobial efficiency via agar diffusion and microbroth dilution methods. The antimicrobial activity was assessed for eight bacterial (Bacillus cereus, Staphylococcus aureus, Salmonella enterica subsp. enterica, Proteus vulgaris, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Listeria monocytogenes), five moulds (Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus, Penicillium citrinum, Penicillium expansum) and one yeast strain (Saccharomyces cerevisiae). The agar diffusion method revealed the blackberry GTE as the most effective since it inhibited the growth of three bacterial, four moulds and one yeast species, having considered the total number of affected microorganism species. Next to the blackberry, the olive GTE appeared to be the second most efficient, suppressing five bacterial strains but no moulds or yeasts. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined for each GTE and the microorganisms tested. Noticeably, the olive GTE appeared to feature the strongest bacteriostatic and bactericidal outcome, displaying specificity for S. aureus, E. faecalis and L. monocytogenes. The other GTEs, such as blueberry, walnut, black mulberry and almond (the list indicates relative strength), were more effective at suppressing microbial growth than inducing microbial death. However, some species specificities were also evident, while the blackcurrant GTE had no significant antimicrobial activity. Having seen the antimicrobial properties of the analysed GTEs, especially the olive and black mulberry GTEs, these could be envisioned as potential antimicrobials that might enhance antibiotic therapies efficiency, while the blackberry GTE would act as an antifungal agent. Some of the GTE mixtures analysed have shown interesting antimicrobial synergies, and all the antimicrobial effects observed argue for extending these studies to include pathological microorganisms.

Biocide Tolerance and Impact of Sanitizer Concentrations on the Antibiotic Resistance of Bacteria Originating from Cheese.

In this study, we determined and identified the bacterial diversity of different types of artisanal and industrially produced cheese. The antibiotic (erythromycin, chloramphenicol, kanamycin, ampicillin, clindamycin, streptomycin, tetracycline, and gentamicin) and biocide (peracetic acid, sodium hypochlorite, and benzalkonium chloride) resistance of clinically relevant bacteria was determined as follows: Staphylococcus aureus, Macrococcus caseolyticus, Bacillus sp., Kocuria varians, Escherichia coli, Enterococcus faecalis, Citrobacter freundii, Citrobacter pasteurii, Klebsiella oxytoca, Klebsiella michiganensis, Enterobacter sp., Enterobacter cloacae, Enterobacter sichuanensis, Raoultella ornithinolytica, Shigella flexneri, and Salmonella enterica. Also, the effect of the sub-inhibitory concentration of three biocides on antibiotic resistance was determined. The microbiota of evaluated dairy products comprise diverse and heterogeneous groups of bacteria with respect to antibiotic and disinfectant tolerance. The results indicated that resistance was common in the case of ampicillin, chloramphenicol, erythromycin, and streptomycin. Bacillus sp. SCSSZT2/3, Enterococcus faecalis SRGT/1, E. coli SAT/1, Raoultella ornithinolytica MTT/5, and S. aureus SIJ/2 showed resistance to most antibiotics. The tested bacteria showed sensitivity to peracetic acid and a different level of tolerance to benzalkonium chloride and sodium hypochlorite. The inhibition zone diameter of antibiotics against Enterococcus faecalis SZT/2, S. aureus JS11, E. coli CSKO2, and Kocuria varians GRT/10 was affected only by the sub-inhibitory concentration of peracetic acid.

Antibiotic resistance pattern of the allochthonous bacteria isolated from commercially available spices.

Spices are often used in dried form, sometimes with significant microbial contamination including pathogenic and food spoilage bacteria. The antibiotic resistance represents an additional risk for food industry, and it is worthy of special attention as spices are important food additives. During our work, we examined the microbiological quality of 50 different spices with cultivation methods on diverse selective media. The identification of the most representative bacteria was carried out using 16S rDNA gene sequence analysis. Antibiotic resistance profiling of twelve identified Bacillus species (B. subtilis subsp. stercoris BCFK, B. licheniformis BCLS, B. siamensis SZBC, B. zhangzhouensis BCTA, B. altitudinis SALKÖ, B. velezensis CVBC, B. cereus SALÖB isolate, B. tequilensis KOPS, B. filamentosus BMBC, B. subtilis subsp. subtilis PRBC2, B. safensis BMPS, and B. mojavensis BCFK2 isolate) was performed using the standard disk-diffusion method against 32 antibiotics. The study showed that the majority resistance was obtained against penicillin G (100%), oxacillin (91.67%), amoxyclav (91.67%), rifampicin (75%), and azithromycin (75%). Our findings suggest that spices harbor multidrug-resistant bacteria.

The effect of essential oils and their combinations on bacteria from the surface of fresh vegetables.

During the study, we determined the antimicrobial activity of different selected essential oils (thyme, lemongrass, juniper, oregano, sage, fennel, rosemary, mint, rosehips, dill) on some pathogenic and spoilage bacteria isolated from the surface of various fresh vegetables. At the same time, in the case of some volatile oil combinations we followed the phenomena of synergism and antagonism. The identification of the isolated bacterial strains was made using 16S rDNA gene sequence analysis. The most resistant isolates appeared to be Curtobacterium herbarum, Achromobacter xylosoxidans, and Enterobacter ludwigii, while Pseudomonas hibiscicola was the most sensitive. Of the chosen plant essential oils, the most pronounced antimicrobial effect was detected in the case of oregano. The essential oils of thyme and mint also showed elevated antimicrobial activity. A synergistic effect was observed in case of five combinations of essential oil. Based on the results, we find that some individual essential oils and mixture compositions (due to synergic effect) could be good candidates for the preservation of fresh vegetables. These preliminary findings suggest that essential oils from locally grown spices could contribute to decreasing the health risk and also to the suppression of emergence of antibiotic resistance.

Investigation of mineral water springs of Miercurea Ciuc (Csíkszereda) region (Romania) with cultivation-dependent microbiological methods.

Water samples of ten mineral water springs at Miercurea Ciuc (Csíkszereda) region (Romania) were examined during 2005-2006 using cultivation-dependent microbiological methods. The results of standard hygienic bacteriological tests showed that the Hargita Spring had perfect and five other springs had microbiologically acceptable water quality (Zsögöd-, Nagy-borvíz-, Taploca-, Szentegyháza- and Lobogó springs). The water of Borsáros Spring was exceptionable (high germ count, presence of Enterococcus spp.).Both standard bacteriological and molecular microbiological methods indicated that the microbiological water quality of the Szeltersz-, Nádasszék- and Délo springs was not acceptable. Bad water quality resulted from inadequate spring catchment and hygiene (low yield, lack of runoff, negligent usage of the springs, horse manure around the spring).The 16S rRNA gene-based identification of strains isolated on standard meat-peptone medium resulted in the detection of typical aquatic organisms such as Shewanella baltica, Aeromonas spp., Pseudomonas veronii, Psychrobacter sp,. Acinetobacter spp. and allochthonous microbes, like Nocardia, Streptomyces, Bacillus, Microbacterium , and Arthrobacter strains indicating the impact of soil. Other allochthonous microbes, such as Staphylococcus spp., Micrococcus sp., Lactococcus sp., Clostridium butyricum, Yersinia spp., Aerococcus sp., may have originated from animal/human sources.