Biopreservation potential of Enterococcus faecalis isolated from Italian traditional raw milk cheeses

Enterococcus faecalis is frequently associated with raw milk cheeses of Mediterranean area. The genetic diversity of 38 E. faecalis obtained from raw milk products in Italy was assessed through Randomly Amplified Polymorphic DNA PCR (RAPD-PCR) and repetitive extragenic palindromic PCR (rep-PCR). The strains were screened for their antimicrobial activity against 5 food-borne spoilage and pathogenic bacteria and 13 lactic acid bacteria (LAB), commonly used as starter cultures. Investigation was made to identify the bacteriocinogenic potential by searching for bacteriocin-encoding genes. Inhibitory effects against undesirable bacteria, including Bacillus cereus (44.7% of strains), Escherichia coli (18.4%), Listeria monocytogenes (15.8%), Staphylococcus aureus (2.6%), and Clostridium sporogenes (21.1%), were detected. Moderate antagonism towards LAB was found. One strain producing enterocin AS-48 was identified, suggesting that the antimicrobial activity of the phenotypically positive isolates should be neces..

Molecular typing of Staphylococcus pseudintermedius canine strains by three commonly used techniques

Staphylococcus pseudintermedius is a newly described species of Staphylococcus regarded as the main causative agent of canine pyoderma [1]. S. pseudintermedius infection was recently described in humans. An important feature of this pathogen is the high genetic identity with two other species of staphylococci, namely S. intermedius and S. delphini, which are included all together in the Staphylococcus Intermedius Group (SIG) [2]. This scenario seriously hampers phenotypic differentiation of these three pathogens. Despite this, only in 2008 was described the first molecular protocol for diagnostic identification of   S. pseudintermedius [3]. The aim of this work was to investigate the presence of different biotypes of S. pseudintermedius obtained from clinically relevant cases of pyoderma in dogs using three molecular methods commonly used to type bacteria: the Ribosomal Spacers Amplification (RSA), the Random Amplification of Polymorphic DNA (RAPD) and the Restriction Fragment Length Polymorphism (RFLP). A total of 46 different strains were included in this work. The application of the RSA technique, which was applied here for the first time, identified the presence of S. pseudintermedius, although it did not allow any differentiation between biotypes. The RAPD assay showed a single cluster that assembles all the interested strains that are grouped in three different sub-clusters (Fig. 1). The RFLP technique showed the most discriminative power, providing the opportunity to clearly identify this bacterium. In conclusion, the use of these three different techniques allows to clearly identify S. pseudintermedius and to observe the presence of different biotypes. In future it could be interesting to couple these results with the determination of the antibiotic resistance in order to verify if certain Multi Drug Resistant strains have particular RSA and RAPD profiles

Efficiency of cleaning procedure of milking equipment and bacterial quality of milk

The cleaning and sanitation of milking equipment could be consider a critical point in the milking procedure because a cleaning failure could influence the level of bacteria contamination of bulk tank milk. Aim of the study is to monitor the cleaning procedures of milking systems in 7 dairy cows farms in Lombardy and to find a relation between efficacy of cleaning system and the bacterial quality of bulk tank milk, remaining washing water through milking equipment and teat cup surface. Cleaning procedures were monitored with Lactocorder, that measured: duration, water temperature, turbulence, percentage of water in pipes, water conductivity of pre- and post-rinse and detergent phases. Results showed that the monitored farms the most of the cleaning parameters were lower than recommendations, in particular maximum water temperature (42.1±9.9°C) and percentage of water during detergent phase (76.1±13.9 %). A maximum temperature of detergent phase <40°C determined a high Standard Plate Count (SPC), thermoduric bacteria and Coliform Count (CC) of bulk tank milk, SPC and CC of teat cup surface. The research indicated that monitoring the efficiency of cleaning milking equipment with proper tools provide useful information about possible sources of contamination of bulk tank milk. Improving cleaning milking efficiency allow to improve milk quality

Phenotypic and Genotypic Characterization of Staphylococcus aureus Strains from Italian Dairy Products

Staphylococcus aureus is a known major cause of foodborne illnesses, and milk and dairy products are often contaminated by enterotoxigenic strains of this bacterium. In the present study, 122 S. aureus isolates collected from different dairy products were characterised by phenotypic properties, by the distribution of genes encoding staphylococcal enterotoxins (sea, sec, sed, seg, seh, sei, sej, and sel) and by randomly amplified polymorphic DNA PCR (RAPD-PCR). Moreover, strain resistance to vancomycin and methicillin (oxacillin) was studied. The differences in the RAPD-PCR profiles obtained with the primers M13 and AP4 revealed the presence of a great genetic heterogeneity among the different S. aureus strains. Using the primer AP4 and M13, eight groups were distinguished by RAPD-PCR cluster analysis, although, except in few cases, it was not possible to correlate the isolates of different animal species (cow or ovine) with the presence of se genes. None of the isolates showed resistance to vancomycin or methicillin

Lactic acid bacteria with cholesterol-lowering properties for dairy applications: In vitro and in situ activity.

Cholesterol-lowering activity is one of the most promising properties of lactic acid bacteria with probiotic characteristics. In the present study, 58 potentially probiotic lactic acid bacteria were tested for their ability to survive in vitro digestion and reduce cholesterol in a medium containing cholesterol and bile acids. The best-performing strains (Lactobacillus casei VC199, Lactobacillus paracasei ssp. paracasei SE160 and VC213, Lactobacillus plantarum VS166 and VS513, Enterococcus faecium VC223, and Enterococcus lactis BT161) resulted in a 42 to 55% reduction of the cholesterol level in broth and were further tested in cheese manufacture. The cholesterol content in all the cheeses decreased with ripening. All the strains were present in the cheese at levels higher than 107 cfu/g until 60 d of ripening, the highest reductions (up to 23%) being obtained when Lb. paracasei ssp. paracasei VC213 and E. lactis BT161 were added during the cheese-making. The adjunct cultures had no negative effect on the sensory characteristics of the cheese. Thus, these strains with proven in vitro properties are good candidates for novel probiotic-containing formulations and could be used to functionalize foods such as dairy fermented products

Effects of the Cooling Temperature at the Farm on Milk Maturation and Cheesemaking Process in the Manufacture of Parmigiano Reggiano PDO Cheese

Parmigiano Reggiano is a hard PDO cheese made from bovine raw milk, whose microbiological characteristics have important repercussions on cheese quality. According to the EU official production protocol, milk temperature at the farm must not drop below 18 °C. The present research aimed to study the effect of cooling milk at the farm at 9 °C on the characteristics of milk and on the cheesemaking process and losses during manufacture. Six cheesemaking trials were performed in two different dairies. In each of them, two cheesemakings were made in parallel: one with milk kept at 9 °C (TM9) and the other with milk kept at 20 °C (TM20). TM9 milk, in comparison with TM20, showed after the creaming process a significant reduction not only of total bacterial count but also of psychrotrophic and lipolytic bacteria. At the same time, TM9 milk showed a higher creaming capacity and, consequently, a lower fat content than TM20. TM9 vat milk had worst coagulation properties than TM20, which caused slightly higher loss of fat and curd fines into the whey. Nevertheless, these changes were too small to influence the efficiency of the cheesemaking process; conversely, maintaining milk at the farm at 9 °C led to a reduction of the number of spoilage bacteria

Characterisation of Formaggella della Valle di Scalve Cheese Produced From Cows Reared in Valley Floor Stall or in Mountain Pasture: Fatty Acids Profile and Sensory Properties

An important problem in mountain areas is the abandonment of pasture. This trend can be combated by the valorisation of typical dairy products, such as "Formaggella della Valle di Scalve", a semi-cooked traditional cheese made from whole milk in a mountain area in Italy. The aim of the present research was to compare the fatty acid (FA) profile and the sensory properties of this cheese as manufactured under different conditions: i) from the milk of cows grazing on mountain or valley pasture or fed indoors; ii) from the milk of cows fed hay or fed silage. In the first case, five cheesemaking trials were conducted during two years for each of the following situations: mountain pasture (A); pasture at the bottom of the valley (P) (about 1000m asl); stall (S). In the second case, three cheesemaking trials were conducted for each of the following situations: cows fed silage (I); cows fed hay (F). S cheese was richer in medium-chain FAs, while long-chain FAs were higher in P and A cheeses. On the other hand, long chain fatty acids (LCFA) were more abundant in P and A cheeses than in S. In general, MUFA, PUFA and, consequently, total unsaturated FA (UFA), were significantly higher in the P and A cheeses than S (UFA: 36.55 and 38.34, respectively, vs 31.13; p &lt; 0.001), while SFA showed higher values in S (68.85 vs 63.41 and 61.68 in P and A, respectively; p &lt; 0.001). Conjugated linoleic acid isomers (CLA) were more represented in the P and A samples (1.86 in P and 1.52 in A, vs 0.80 in S; p &lt; 0.001); Omega 3 fatty acids, and in particular α-linolenic acid, were more abundant in P than in S cheese. In winter, the I sample (silage) presented higher percentages of myristic (C14), myristoleic (C14:1) and omega 6 acids, whereas F cheese (hay) contained higher concentrations of CLA. The triangular test of sensory analysis showed that, in general, F cheeses were judged as "sweeter" than I, with aromatic profiles characterized by higher content of 2- butanol and ethyl capronate

Effect of a reduced amount of straw bedding on goats' comfort and hygienic characteristics of milk and straw

The present study aims to understand the influence of a reduced amount of straw litter on goats' lying comfort and hygienic characteristics of milk and straw. Lying behaviour (frequency, lying post..

Bacterial Communities in the Embryo of Maize Landraces:Relation with Susceptibility to Fusarium Ear Rot

Locally adapted maize accessions (landraces) represent an untapped resource of nutritional and resistance traits for breeding, including the shaping of distinct microbiota. Our study focused on five different maize landraces and a reference commercial hybrid, showing different susceptibility to fusarium ear rot, and whether this trait could be related to particular compositions of the bacterial microbiota in the embryo, using different approaches. Our cultivation-independent approach utilized the metabarcoding of a portion of the 16S rRNA gene to study bacterial populations in these samples. Multivariate statistical analyses indicated that the microbiota of the embryos of the accessions grouped in two different clusters: one comprising three landraces and the hybrid, one including the remaining two landraces, which showed a lower susceptibility to fusarium ear rot in field. The main discriminant between these clusters was the frequency of Firmicutes, higher in the second cluster, and this abundance was confirmed by quantification through digital PCR. The cultivation-dependent approach allowed the isolation of 70 bacterial strains, mostly Firmicutes. In vivo assays allowed the identification of five candidate biocontrol strains against fusarium ear rot. Our data revealed novel insights into the role of the maize embryo microbiota and set the stage for further studies aimed at integrating this knowledge into plant breeding programs

Caracterización de Staphylococcus aureus aislado de productos lácteos por MALDI-TOF MS

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