The emergence of antibiotic-resistant bacteria has become a major concern worldwide. This trend indicates the need for alternative agents to antibiotics, such as natural compounds of plant origin. Using agar disc diffusion and minimum inhibitory concentration (MIC) assays, we investigated the antimicrobial activity of Citrus aurantium (AEO), Citrus x limon (LEO), Eucalyptus globulus (EEO), Melaleuca alternifolia (TTO), and Cupressus sempervirens (CEO) essential oils (EOs) against three representatives of antibiotic-resistant pathogens and respective biofilms: vancomycin-resistant enterococci (VRE), methicillin-resistant Staphylococcus aureus (MRSA), and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli. Using the checkerboard method, the efficacy of the EOs alone, in an association with each other, or in combination with the reference antibiotics was quantified by calculating fractional inhibitory concentrations (FICs). All the EOs displayed antibacterial activity against all strains to different extents, and TTO was the most effective. The results of the EO–EO associations and EO–antibiotic combinations clearly showed a synergistic outcome in most tests. Lastly, the effectiveness of EOs both alone and in association or combination against biofilm formed by the antibiotic-resistant strains was comparable to, and sometimes better than, that of the reference antibiotics. In conclusion, the combination of EOs and antibiotics represents a promising therapeutic strategy against antibiotic-resistant bacteria, even protected inside biofilms, which can allow decreasing the concentrations of antibiotics used.
Probiotics retrieved from animal sources have substantial health benefits for both humans and animals. The present study was designed to identify lactic acid bacteria (LAB) isolated from domestic water buffalo milk (Bubalus bubalis) and to evaluate their potential as target-based probiotics. Forty-six LAB strains were isolated and, among them, five strains (NMCC-M2, NMCC-M4, NMCC-M5, NMCC-M6, and NMCC-M7) were regarded as possible probiotics on the basis of their phenotypic and biochemical properties. These isolates were molecularly identified as Weissella confusa (NMCC-M2), Leuconostoc pseudo-mesenteroides (NMCC-M4), Lactococcus lactis Subsp. hordniae (NMCC-M5), Enterococcus faecium NMCC-M6, and Enterococcus lactis NMCC-M7. The tested bacterial strains showed significant antimicrobial activity, susceptibility to antibiotics, acid and bile tolerance, sugar fermentation, enzymatic potential, and nonhemolytic characteristics. Interestingly, NMCC-M2 displayed the best probiotic features including survival at pH 3 and 0.5% (w/v) bile salts, complete susceptibility to the tested antibiotics, high enzymatic potential, and in vitro cholesterol reduction (48.0 µg/mL for NMCC-M2) with 0.3% bile salt supplementation. Therefore, the isolated strain NMCC-M2 could be considered as a potential target-based probiotic in cholesterol-lowering fermented food products.
The prevalence of multi-drug-resistant (MDR) bacteria in 48 air samples from indoor environments, surgical rooms, dental surgery and waste management plants has been investigated. A total of 280 bacterial strains belonging to different genera were isolated, and the operating rooms were the most contaminated ones (107 isolates), with all the isolates belonging to Gram-positive cocci (51.5 % Micrococcus spp., 48.5 % Staphylococcus spp.). Only 5 % of the isolates was sensitive to all the antibiotics tested, while the remaining strains resulted resistant to three (13 %), four (14 %), five (9 %) and six (10 %) antibiotics. Correlation between the resistance patterns and the environmental source of MDR bacteria isolates also emerged from the present investigation. This study confirms the high presence of antibiotic-resistant bacteria in air samples, finding that represents a threat for the possible transfer of resistance genes to pathogenic bacteria
Cockroaches, insects of the order Blattodea, seem to play a crucial role in the possible conjugation-mediated genetic exchanges that occur among bacteria that harbor in the cockroach intestinal tract. The gut of these insects can be thought of as an effective in vivo model for the natural transfer of antimicrobial resistance plasmids among bacteria. In our study, we evaluated the conjugation-mediated horizontal transfer of resistance genes between Escherichia coli and other microorganisms of the same Enterobacteriaceae family within the intestinal tract of Blaberus craniifer Burmeister, 1838 (Blattodea: Blaberidae). Different in vivo mating experiments were performed using E. coli RP4 harboring the RP4 plasmid carrying ampicillin, kanamycin, and tetracycline resistance genes as the donor and E. coli K12 resistant to nalidixic acid or Salmonella enterica serovar Enteritidis IMM39 resistant to streptomycin as the recipients. The RP4 plasmid was successfully transferred to both recipients, producing E. coli K12-RP4 and S. Enteritidis IMM39-RP4 transconjugants. Conjugation frequencies in vivo were similar to those previously observed in vitro. The transfer of the RP4 plasmid in all transconjugants was confirmed by small-scale plasmid isolation and agar gel electrophoresis, suggesting that the intestinal tract of cockroaches is an effective in vivo model for natural gene transfer. Our results confirm that cockroaches allow for the exchange of antimicrobial resistance plasmids among bacteria and may represent a potential reservoir for the dissemination of antibiotic-resistant bacteria in different environments. These findings are particularly significant to human health in the context of health care settings such as hospitals.
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