Università degli studi di Modena e Reggio Emilia

The anti-Listeria monocytogenes activity of four essential oils (EOs) (Salvia officinalis, Citrus limon, Mentha piperita and Thymus vulgaris) and bacteriocin bacLP17, added alone or in mixture in active edible coatings, was determined in artificially contaminated shrimps. The minimal inhibitory concentration (MIC) values of the EOs were determined against the NCTC 10888 strain of L. monocytogenes by using the broth microdilution method. The checkerboard method was carried out in tryptic soy broth (TSB), using microdilution to obtain the Fractional Inhibitory Concentration Index (FIC-Index) for six associations of EOs, chosen based on the best MIC results. All the EOs confirmed their anti-Listeria activity, both “in vitro” and inside the coatings. The coating matrix was suitable for use in the food field, allowing a gradual release of the EOs in packaged food. When the EOs were used in association (EO/EO) they were demonstrated to act synergistically, leading to a significant reduction in the amount (10–20 times) of EOs needing to be used, and consequently a decrease in the strong smell on the food. This effect was also confirmed when the compounds were incorporated into the coatings. The inclusion of the EOs within the coating not only ensured the anti-Listeria activity by increasing the shelf-life of food products, but also further mitigated the strong smell of the EOs, improving the organoleptic impact on the food and its sensory properties. Keywords: Listeria monocytogenes; shrimp; essential oils; bacteriocin; edible coating.

The antimicrobial activity of garlic (Allium sativum L.) and onion (Allium cepa L.) plant active extracts was determined against Listeria monocytogenes in two meat products. Samples of sausages “cacciatore” and cooked ham in vacuum-packaged slices were artificially contaminated, and the presence of Listeria was evaluated during the sausages ripening and throughout the shelf-life of the cooked ham. The test carried out on sausages did not show differences among treated and untreated samples. The antagonistic activity of the plant extracts against the pathogen was probably hidden by the competition from the sausages microbial flora and the pH and the water activity (aw) decrease. On the other hand, the plant extracts determined an initial reduction of about 1.00 log cfu/g of the L. monocytogenes viable count in the cooked ham slices contaminated with 103 cfu/g, but the best result was obtained with the contamination of 102 cfu/g of L. monocytogenes. In addition to the pathogen’s initial decrease, we observed an extension of the lag phase and a reduction of the Listeria growth rate. Considering that the presence of L. monocytogenes during the slicing phase of the cooked ham does not exceed 10 cfu/g, the use of plant extracts can lead to complete pathogen elimination.

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.

Sixty-nine Lactic Acid Bacteria (LAB) and bifidobacteria were isolated and identified from Italian dairy products (raw milk, cream, butter, soft cheese and yoghurt) to find new antimicrobial compounds to use as food bio-preservatives. All the isolates were preliminarily screened by the deferred antagonism method for bacteriocin production. Afterwards, to evaluate the release of bacteriocin in liquid medium, the Cell-Free Supernatant Fluid (CFSF) of the best producers was tested by agar well diusion assay. The study allowed the selection of three bacteriocin producing strains (Enterococcus faecium E23, Bifidobacterium thermophilum B23 and Lactobacillus bulgaricus L21), endowed with the strongest and broadest inhibitory capability against the pathogen Listeria monocytogenes. The molecular characteristics and the chemical–physical properties of both producers and the respective bacteriocins were studied and compared. The results showed that E. faecium E23 was the best producer strain and its class IIa bacteriocins, called enterocin E23, exhibited a good spectrum of activity towards L. monocytogenes. Enterocin E23 was stable over a wide range of pH and at low temperatures for at least four months and, for this reason, it can be employed in refrigerated foods for the control of L. monocytogenes, the major concern in dairy products.

The aim of this study was to analyze the antibacterial activity of four essential oils (EOs), Melaleuca alternifolia, Eucalyptus globulus, Mentha piperita, and Thymus vulgaris, in preventing the development and spread of extended-spectrum -lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae, metallo-beta-lactamase (MBL)-producing Pseudomonas aeruginosa and carbapenemase (KPC)-producing Klebsiella pneumoniae. A total of 60 strains were obtained from the stock collection from the Microbiology Laboratory of Hesperia Hospital, Modena, Italy. Twenty ESBLproducing E. coli, 5 K. pneumoniae, 13 KPC-producing K. pneumoniae, and 20 MBL-producing P. aeruginosa were cultured and reconfirmed as ESBL and carbapenamase producers. Polymerase chain reaction was used for the detection of genes responsible for antibiotic resistance (ESBL and KPC/MBL). Antibacterial activity of the EOs was determined using the agar disk diusion assay, and minimal inhibitory concentrations (MICs) were also evaluated. Lastly, adhesion capability and biofilm formation on polystyrene and glass surfaces were studied in 24 randomly selected strains. M. alternifolia and T. vulgaris EOs showed the best antibacterial activity against all tested strains and, as revealed by agar disk diusion assay, M. alternifolia was the most eective, even at low concentrations. This eect was also confirmed by MICs, with values ranging from 0.5 to 16 g/mL and from 1 to 16 g/mL, for M. alternifolia and T. vulgaris EOs, respectively. The EOs’ antibacterial activity compared to antibiotics confirmed M. alternifolia EO as the best antibacterial agent. T. vulgaris EO also showed a good antibacterial activity with MICs lower than both reference antibiotics. Lastly, a significant anti-biofilm activity was observed for the two EOs (*P < 0.05 and **P < 0.01 for M. alternifolia and T. vulgaris EOs, respectively). A good antibacterial and anti-biofilm activity of M. alternifolia and T. vulgaris EOs against all selected strains was observed, thus demonstrating a future possible use of these EOs to treat infections caused by ESBL/carbapenemase-producing strains, even in association with antibiotics.

Methodology. Biochemical and molecular methods were used to identify 100 lactobacilli isolated from rectal swabs. Among these, L. paracasei ssp. paracasei LP5 and L. brevis LP9 showed significant antibacterial activity against S. agalactiae and L. monocytogenes. Accordingly, characterization of their bacteriocins, BacLP5 and BacLP9, was conducted to obtain information on their kinetic production, sensitivity to chemico-physical parameters and molecular weight. To investigate the possible use of the two Lactobacillus strains as probiotics, their gastrointestinal resistance, cellular adhesiveness and sensitivity to antibiotics were also studied. Results. The obtained data show that BacLP5 and BacLP9 most likely belong to class II bacteriocins and both have a molecular weight of approximately 3 kDa. The production of BacLP5 and BacLP9 started after 4 h (40 and 80 AU ml−1), respectively. Both of the Lactobacillus strains survived gastric and intestinal juices well and showed adhesive capability on HEp-2 cells. Conclusion. Due to their peculiar antimicrobial characteristics, L. paracasei ssp. paracasei LP5 and L. brevis LP9 are suitable for use in the treatment of vaginal disorders, through both oral and transvaginal administration