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In recent years, the search for new compounds with antibacterial activity has drastically increased due to the spread of antibiotic-resistant microorganisms. In this study, we analyzed Cell-Free Supernatant (CFS) from Bacillus siamensis, assessing its potential antimicrobial activity against some of the main pathogenic microorganisms of human interest. To achieve this goal, we exploited the natural antagonism of skin-colonizing bacteria and their ability to produce compounds with antimicrobial activity. Biochemical and molecular methods were used to identify 247 strains isolated from the skin. Among these, we found that CFS from a strain of Bacillus siamensis (that we named CPAY1) showed significant antimicrobial activity against Staphylococcus aureus, Enterococcus faecalis, Streptococcus agalactiae, and Candida spp. In this study, we gathered information on CFS's antimicrobial activity and on its sensitivity to chemical–physical parameters. Time–kill studies were performed; anti-biofilm activity, antibiotic resistance, and plasmid presence were also investigated. The antimicrobial compounds included in the CFS showed resistance to the proteolytic enzymes and were heat stable. The production of antimicrobial compounds started after 4 h of culture (20 AU/mL). CPAY1 CFS showed antimicrobial activity after 7 h of bacteria co-culture. The anti-biofilm activity of the CPAY1 CFS against all the tested strains was also remarkable. B. siamensis CPAY1 did not reveal the presence of a plasmid and showed susceptibility to all the antibiotics tested. [ABSTRACT FROM AUTHOR]

The presence of antibiotic-resistant bacteria has become a major therapeutic priority. This trend indicates the need for alternative agents to antibiotics, such as natural compounds of plant origin. By assessing membrane permeability, we investigated the antimicrobial activity of Melaleuca alternifolia and Eucalyptus globulus essential oils (EOs) against three strains of methicillin-resistant Staphylococcus aureus (MRSA). Using the checkerboard method, the efficacy of single EOs, in association with each other or in combination with oxacillin, was quantified by calculating the fractional inhibitory concentrations (FIC Index). All EOs showed a reduction in bacterial load, an alteration of membrane permeability which leads to an increase in its function, resulting in the release of nucleic acids and proteins. The treatment with EO–oxacillin combinations and associated EO–EO resulted in a synergistic effect in most of the tests performed. EO–EO association showed a high activity in the alteration of the membrane, increasing the permeability to about 80% in all the MRSA strains treated. In conclusion, the combination of EOs and antibiotics represents a valid therapeutic support against MRSA bacteria, allowing for a decrease in the antibiotic concentration needed for therapeutic use. [ABSTRACT FROM AUTHOR]

Featured Application: Natural compound extracts from lichens were able to decrease and/or inhibit the growth of microorganisms of interest in human health, thus providing a good alternative alone or in combination with drugs for therapy. The antimicrobial properties of two lichen extracts (LC1 and LC2 solutions extracted in acetone and cyclohexane, respectively) were investigated against both Gram-positive and Gram-negative microorganisms through the agar well diffusion assay. Results displayed that both samples were similarly effective against all the indicator strains. The antimicrobial activity was maintained up to 30 days against Candida albicans ATCC 10231 with an inhibition zone of 38 mm and 37 mm for the LC1 and LC2 solutions extracted, respectively. In order to separate the single chemical components and to associate them with the biological activity, the two extracts were subjected to an activity-guided fractionation followed by a liquid chromatography mass spectrometry (LC–MS) Ion Trap 6310A for the chemical characterization. Chromatogram analysis of each sample that maintained an antimicrobial activity revealed the presence of a significant peak, at a retention time (tR) of 10.8 min, corresponding to a scabrosin derivative that could likely be associated with the antimicrobial activity. Results obtained in the present investigation, especially against the opportunistic pathogen C. albicans, are encouraging and could represent a preliminary step to a future solution toward a microorganism responsible for fungal infections, mainly occurring in immunocompromised patients and recently caused by drug-resistant strains. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate and characterize the bacterial load present in twenty-four Ready-To-Eat (RTE) sandwiches, purchased at refrigerated vending machines and supermarkets in the province of Modena (Italy). We isolated 54 bacterial strains, including pathogens of interest in food safety, such as Listeria, Staphylococcus, Enterococcus, Yersinia, Aeromonas and Acinetobacter spp. Phenotypic tests have been performed on these pathogens to detect the presence of virulence factors, such as gelatinase production and hemolytic capability. To test their antibiotic resistance features, the minimum inhibitory concentration (MIC) against eight commonly used antibiotics (Amikacin, Ciprofloxacin, Ampicillin, Oxacillin, Imipenem, Tetracycline, Erythromycin and Vancomycin) was also evaluated. The results showed that among the 54 isolates, fifty percent (50%) belonged to harmless microorganisms (Leuconostoc and Lactococcus), whereas the remaining fifty percent (50%) included pathogenic bacteria (Listeria ivanovii, Listeria monocytogenes, Staphylococcus aureus, Yersinia, and Citrobacter spp.), species responsible for pathologies often difficult to treat due to the presence of antibiotic resistance features. This study demonstrates the importance of thorough controls, both during the production and marketing of RTE food like sandwiches, to avoid reaching the infectious load and the onset of pathologies, particularly dangerous for old and immunocompromised patients. [ABSTRACT FROM AUTHOR]

Several species of the genus Acanthamoeba cause human diseases. Treatment of infections involves various problems, emphasising the need to develop alternative antiprotozoal agents. We studied the anti-amoebic activity of Essential Oils (EOs), derived from rosemary (Rosmarinus officinalis L.) and cloves (Syzygium aromaticum L. Merr. & Perry), against Acanthamoeba polyphaga strain. The amoebicidal activity of cloves and rosemary EOs was preliminary demonstrated by the morphology change (modifications in the cell shape, the presence of precipitates in the cytoplasm, autophagic vesicles, membrane blends) of the treated trophozoites. The cell-counts, carried out after staining trophozoites with a Trypan blue solution, revealed that both EOs were active in a dose-dependent manner and in relation to the exposure time. This activity was evident after few hours, with encouraging results obtained in particular with cloves EO, able to act at the lower concentrations and after 1 h, probably for its high eugenol content (65.30%). [ABSTRACT FROM AUTHOR]

We investigated the presence of ESBL and AmpC-producing Enterobacteriaceae isolated from 200 rectal swabs of healthy swine and 200 samples of ground pork. Phenotypic testing by using the double synergy differential test (DSDT) for ESBL/AmpC-positive strains was confirmed by PCR and DNA sequence analysis. The localization of beta-lactamase genes was established by conjugation experiments. ESBL and/or AmpC-producing Enterobacteriaceae was found in 52.2% (95/182) of the isolates collected from rectal swabs and 3% (3/100) of isolates obtained from ground pork samples. Polymerase chain reaction and sequencing confirmed the presence of blaTEM-20, blaTEM-34, blaTEM-52, blaCTX-M-1, blaSHV-12, blaTEM-1 +SHV-12, blaTEM-20 +SHV-12, blaCMY-2, blaTEM-1+ CMY-2, blaACC-1 and blaACC-2. The conjugation assays yielded positive results, denoting a plasmid localization of the genes. Practical Applications In this study, the prevalence of ESBL and AmpC-producing Enterobacteriaceae isolated from rectal swabs of healthy swine and samples of ground pork were determined. ESBL/AmpC-positive strains were confirmed by PCR and DNA sequence analysis. The most frequently isolated species was E.coli, among the most common variants detected TEM-type ESBL, TEM-52 was showed. These findings provide new information about the presence of ESBL/AmpC at the farm level and have important implications for assessments of risks of meat contamination during slaughter. [ABSTRACT FROM AUTHOR]

One of the most-used food contact materials is stainless steel (AISI 304L or AISI 316L), owing to its high mechanical strength, cleanability, and corrosion resistance. However, due to the presence of minimal crevices, stainless-steel is subject to microbial contamination with consequent significant reverb on health and industry costs due to the lack of effective reliability of sanitizing agents and procedures. In this study, we evaluated the noncytotoxic effect of an amorphous SiOxCyHz coating deposited on stainless-steel disks and performed a time-course evaluation for four Gram-negative bacteria and four Gram-positive bacteria. A low cytotoxicity of the SiOxCyHz coating was observed; moreover, except for some samples, a five-logarithm decrease was visible after 1 h on coated surfaces without any sanitizing treatment and inoculated with Gram-negative and Gram-positive bacteria. Conversely, a complete bacterial removal was observed after 30 s−1 min application of alcohol and already after 15 s under UVC irradiation against both bacterial groups. Moreover, coating deposition changed the wetting behaviors of treated samples, with contact angles increasing from 90.25° to 113.73°, realizing a transformation from hydrophilicity to hydrophobicity, with tremendous repercussions in various technological applications, including the food industry. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The antibacterial activity of Pimpinella anisum L., Cinnamomum zeylanicum , Syzygium aromaticum , and Cuminum cyminum L. essential oils (EOs) against some common pathogenic microorganisms ( Staphylococcus aureus ATCC 6538, Staphylococcus epidermidis ATCC 14990, Enterococcus faecalis ATCC 29212, Streptococcus pyogenes ATCC 1915, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 27853, Aeromonas hydrophila ATCC 7966, Proteus mirabilis ATCC 10005, Klebsiella pneumoniae ATCC 13883, and Candida albicans ATCC 10231) and their biofilms was studied. The EOs inhibitory effects were evaluated by both Agar Well Diffusion assay and Minimum Inhibitory Concentration (MIC) determination. The most active EOs, cinnamon and cloves, were also tested on 18, 24, 48, 72 hours mature biofilms. Cinnamon and cloves exhibited the best results showing a significant activity against all the tested bacteria. Concerning biofilm, results suggest that Cinnamomum zeylanicum oil may be a useful approach to impair the biofilm produced by the tested Gram-negative bacteria. [Formula: see text].

Among several alternatives to control hospital-acquired infections (HAIs), a strategy could be the use of hospital uniforms imbued with antimicrobial substances. For this purpose we evaluated the antibacterial activity of two different silver doped fabrics employed for the production of hospital uniforms. The study was conducted in two-step. In the first the antimicrobial activity was evaluated in vitro against Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 6538, Enterococcus faecalis ATCC 29212. In the second, we tested the total viable counts detected from beginning to end of the work shift on experimental silver doped uniforms worn by doctors, nurses, allied health assistants in different hospital wards. The in vitro tests showed a remarkable antibacterial activity of both silver doped samples (>99.9% reduction within 4h of exposure for Gram-positive and within 24 h for Gram-negative bacteria). The experimental uniforms provided results only slightly in agreement with in vitro data. Even if the increase of total viable counts was somewhat lower for experimental uniforms than traditional ones, significant differences were not observed. Despite the results on the uniforms worn, the addition of silver in fabrics to make medical equipment (supplies) remains an interesting option for HAI control.

We investigated presence and prevalence of antibiotic-resistances and other biological characters in enterococci isolated from faeces of healthy dogs and cats because these microorganisms represent important human and veterinary pathogens/opportunists, and a significant burden for healthcare systems. In all samples (n=115) we detected enterococci, with a predominance of Enterococcus faecium (42; 36.5%) and Enterococcus faecalis (36; 31.3%) species, endowed with virulence traits and multidrug-resistance. The two predominant resistance patterns (erythromycin, tetracycline) were examined by polymerase chain reaction for tet and erm genes. Only tetM for tetracycline, and ermA and ermB for erythromycin were detected. PCR for gelatinase gene (gelE) was positive in 62.6% of isolates, but only 26.1% produce gelatinase suggesting the existence of silent genes. efaAfs and efaAfm genes were found in E. faecalis and E. faecium respectively. 89.6% of isolates produced bacteriocin-like substances with a prevailing action against Listeria genus and, among these, 33.9% were positive for the bacteriocin structural genes entA, entL50 or entP. According to our study, pet animals can be considered a reservoir of potentially pathogenic enterococci and we cannot exclude that those microorganisms may be responsible for opportunistic infections in high-risk pet owners.

The aim of this study was to assess the production of extended spectrum beta-lactamases (ESBL) in 56 strains of Enterobacteriaceae, obtained from 100 rectal swabs of farm animals, and to evaluate the horizontal transfer ca- pacity of the genetic determinants of resistance. The ESBL-positive strains were confirmed by phenotypic testing, confirmed by PCR and DNA sequence analysis. The localization of beta-lactamase genes was established by conju- gation experiments. Of the 56 analyzed strains, 20 (36%) resulted positive for ESBL production by the double-disk synergy test, and belonged to Escherichia coli 15 (75%) and Klebsiella ozaenae 5 (25%) species. Molecular analysis showed that all ESBL-producing isolates possessed genes encoding for TEM-type enzymes and/or CTX-M. The conjugation assays yielded positive results, thus denoting a plasmidic localization of the genes. This study high- lights the high percentage of ESBL-positive Enterobacteriaceae and the mobility of the responsible genes. Gene mo- bility implies highly negative consequences in terms of drug therapy because of the spread of antibiotic resistance.