Effects of aminomethylphosphonic acid, the main breakdown product of glyphosate, on cellular and biochemical parameters of the mussel Mytilus galloprovincialis.

The effects of the breakdown products of herbicides on aquatic species are largely unknown. In a recent study, we evaluated the effects of glyphosate on the mussel Mytilus galloprovincialis. This study was designed to evaluate for the first time the impact of aminomethylphosphonic acid (AMPA) - the main breakdown product of glyphosate - on cellular and biochemical parameters of the mussel Mytilus galloprovincialis. Bivalves were exposed for 7, 14 and 21 days to 1, 10 and 100 µg/L of AMPA and various biomarkers were measured in haemolymph (total haemocyte counts, haemocyte diameter and volume, haemolymph pH, haemocyte proliferation, haemolymph lactate dehydrogenase activity, haemocyte lysate lysozyme and acid phosphatase activities), as well as in gills and digestive gland (superoxide dismutase, catalase, glutathione S-transferase and acetylcholinesterase activities). AMPA concentrations in seawater samples from the experimental tanks were also measured in order to correlate the biomarker responses of mussels with their exposure to the actual concentrations of AMPA. The MANOVA analysis demonstrated that the experimental variables considered (exposure dose, exposure duration, and their interaction) affected significantly biomarker responses. Nevertheless, the two-way ANOVA analysis revealed significant effects of AMPA on most of the biomarkers measured. The overall results of this study demonstrated that AMPA can affect cellular and biochemical parameters in mussels, similarly to glyphosate.

Inhibitory Effect of Lactiplantibacillusplantarum and Lactococcus lactis Autochtonous Strains against Listeria monocytogenes in a Laboratory Cheese Model.

In the present study, six Lactococcus lactis and seven Lactiplantibacillus plantarum strains isolated from artisanal Sardinian dairy products were evaluated for their efficacy in controlling the growth of Listeria monocytogenes during the storage of miniature fresh cheese manufactured on a laboratory scale to exploit their possible use as biopreservatives. The strains were tested for antimicrobial activity and some technological characteristics before using them in miniature fresh cheese to evaluate their in situ antilisterial effect. Our results showed that five strains (L. lactis 16FS16-9/20234-11FS16 and Lpb. plantarum 1/14537-4A/20045) could be considered suitable candidates for use as protective cultures in fresh cheese manufacture since they significantly lowered the pathogen counts by 3-4 log units compared to the control; however, all strains tested were capable of decreasing L. monocytogenes numbers. Our results suggest that the single and combined action of the acidifying power and the production of bacteriocin of these strains was capable of controlling and/or reducing the growth of L. monocytogenes. Considering their technological characteristics, they might be used as starter/adjunct cultures to increase the safety of the products, perhaps in association with other antimicrobial hurdles.

Influence of Autochthonous Putative Probiotic Cultures on Microbiota, Lipid Components and Metabolome of Caciotta Cheese.

The present study was undertaken to produce probiotic Caciotta cheeses from pasteurized ewes' milk by using different combinations of autochthonous microbial cultures, containing putative probiotic strains, and evaluate their influence on gross composition, lipid components, sensory properties and microbiological and metabolite profiles of the cheeses throughout ripening process. A control cheese was produced using commercial starter cultures. The hydrophilic molecular pools (mainly composed by amino acids, organic acids, and carbohydrates) were characterized by means of 1H NMR spectroscopy, while the cholesterol, α-tocopherol and fatty acid composition by HPLC-DAD/ELSD techniques. Conventional culturing and a PCR-DGGE approach using total cheese DNA extracts were used to analyze cheese microbiota and monitor the presence and viability of starters and probiotic strains. Our findings showed no marked differences for gross composition, total lipids, total cholesterol, and fatty acid levels among all cheeses during ripening. Differently, the multivariate statistical analysis of NMR data highlighted significant variations in the cheese' profiles both in terms of maturation time and strains combination. The use of autochthonous cultures and adjunct probiotic strains did not adversely affect acceptability of the cheeses. Higher levels of lactobacilli (viability of 108-109 cfu/g of cheese) were detected in cheeses made with the addition of probiotic autochthonous strains with respect to control cheese during the whole ripening period, suggesting the adequacy of Caciotta cheese as a carrier for probiotic bacteria delivery.

Application of Autochthonous Lactobacillus Strains as Biopreservatives to Control Fungal Spoilage in Caciotta Cheese.

Fungal spoilage is one of the main causes of economic losses worldwide in the food industry. In the last years, consumer's demands for preservative-free processed foods have increased as a result of growing awareness about the health hazards associated with chemicals. Lactic acid bacteria have been extensively studied for their antibacterial and antifungal potential in order to be used as biopreservatives. The first objective of this study was to investigate in vitro the antifungal activity of autochthonous Lactobacillus strains against moulds commonly associated with cheese spoilage. Then, the Lactobacillus strains with the highest inhibitory effect and broadest spectrum were tested in single or mixed cultures against Penicillium chrysogenum ATCC 9179 and Aspergillus flavus ATCC 46283 on miniature Caciotta cheese produced at laboratory scale to evaluate in situ their ability to prevent mould growth and to determine their impact on cheese organoleptic properties and starter culture activity. The growth of the starter lactococcal population exhibited similar trend and values during ripening, suggesting that the addition of lactobacilli did not influence its growth and survival. Inhibition of P. chrysogenum inoculated in the milk was determined in cheeses produced with single or mixed Lactobacillus adjuncts as compared to cheeses without adjunct. The mixed adjunct cultures resulted in more effective, significantly reducing mould counts of more than 2 log units at the end of ripening. The application of the adjunct cultures resulted in a delay in mycelial growth of P. chrysogenum and A. flavus inoculated on the cheese surface as well. Finally, we found no significant differences among samples for the sensory parameters evaluated that received similar ratings. Our results indicate that the selected Lactobacillus strains may have a potential effect in controlling mould contamination on cheeses. Further studies are currently being carried out to identify the molecules responsible for the antifungal activity.

Biological Activities of Aerial Parts Extracts of Euphorbia characias.

The aim of the present study was to evaluate antioxidant, antimicrobial, anti-HIV, and cholinesterase inhibitory activities of aqueous and alcoholic extracts from leaves, stems, and flowers of Euphorbia characias. The extracts showed a high antioxidant activity and were a good source of total polyphenols and flavonoids. Ethanolic extracts from leaves and flowers displayed the highest inhibitory activity against acetylcholinesterase and butyrylcholinesterase, showing potential properties against Alzheimer's disease. Antimicrobial assay showed that leaves and flowers extracts were active against all Gram-positive bacteria tested. The ethanolic leaves extract appeared to have the strongest antibacterial activity against Bacillus cereus with MIC value of 312.5 µg/mL followed by Listeria monocytogenes and Staphylococcus aureus that also exhibited good sensitivity with MIC values of 1250 µg/mL. Moreover, all the extracts possessed anti-HIV activity. The ethanolic flower extract was the most potent inhibitor of HIV-1 RT DNA polymerase RNA-dependent and Ribonuclease H with IC50 values of 0.26 and 0.33 µg/mL, respectively. The LC-DAD metabolic profile showed that ethanolic leaves extract contains high levels of quercetin derivatives. This study suggests that Euphorbia characias extracts represent a good source of natural bioactive compounds which could be useful for pharmaceutical application as well as in food system for the prevention of the growth of food-borne bacteria and to extend the shelf-life of processed foods.

Preliminary evaluation of probiotic properties of Lactobacillus strains isolated from Sardinian dairy products.

Twenty-three Lactobacillus strains of dairy origin were evaluated for some functional properties relevant to their use as probiotics. A preliminary subtractive screening based on the abilities to inhibit the growth of microbial pathogens and hydrolyze conjugated bile salts was applied, and six strains were selected for further characterization including survival under gastrointestinal environmental conditions, adhesion to gut epithelial tissue, enzymatic activity, and some safety properties. All selected strains maintained elevated cell numbers under conditions simulating passage through the human gastrointestinal tract, well comparable to the values obtained for the probiotic strain Lactobacillus rhamnosus GG, and were able to adhere to Caco-2 cells to various extents (from 3 to 20%). All strains exhibited high aminopeptidase, and absent or very low proteolytic and strong ß-galactosidase activities; none was found to be haemolytic or to produce biogenic amines and all were susceptible to tetracycline, chloramphenicol, erythromycin, ampicillin, and amoxicillin/clavulanic acid. Our results indicate that the Lactobacillus strains analyzed could be considered appropriate probiotic candidates, due to resistance to GIT simulated conditions, antimicrobial activity, adhesion to Caco-2 cell-line, and absence of undesirable properties. They could be used as adjunct cultures for contributing to the quality and health related functional properties of dairy products.

A NMR metabolomics study of the ripening process of the Fiore Sardo cheese produced with autochthonous adjunct cultures.

Fiore Sardo (FS) is a traditional Italian raw ewe's milk cheese carrying a Protected Designation of Origin (PDO). This study investigated the kinetics of FS cheese ripening by physicochemical parameters, microbial counting, and NMR metabolomics using aqueous extracts. Four Fiore Sardo cheeses, manufactured from milk with deliberately added autochthonous lactic acid bacteria (LAB) or commercial starters were studied during a period of 90days of ripening. Major differences in the metabolic profiles were observed amongst the samples as a function of the adjunct culture utilised. (1)H NMR metabolomics in combination with multivariate data analysis was able to classify cheese samples on the basis of their maturation age and the type of added cultures. These findings lay the metabolic basis for the authentication of Fiore Sardo cheese produced in compliance with PDO specifications which allow the use of only native LAB cultures.