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Abstract Background In the context of the RABOLA project, which aimed to identify operational practices that lead to the reduction of antibiotic use in dairy cattle farming, lyophilised Aloe arborescens was administered orally to cows during the dry-off and peripartum periods. In this specific paper we wanted to examine whether oral administration of Aloe arborescens, in combination with the topical application of a teat sealant could exert an effect on the microbial populations of three cow microbiomes (rumen, milk, rectum), between dry-off and peripartum. Dry-off and peripartum are critical physiological phases of the cow’s life, where both the mammary gland and the gastrointestinal tract undergo dramatic modifications, hence the relevance of evaluating the effects of dietary treatments. Methods Thirty multiparous dairy cows were randomly allocated to three groups: Control (antibiotic treatment and internal teat sealant), Sealant (only internal teat sealant) and Aloe (internal teat sealant and Aloe arborescens homogenate administered orally). For 16S rRNA gene sequencing, rumen, rectum and milk samples were collected, not synchronously, at the most critical timepoints around dry-off and calving, considering the physiological activity of each biological site. Results The rumen microbiome was predominantly characterized by Bacteroidetes and Firmicutes followed by Proteobacteria, while the rectum exhibited a prevalence of Firmicutes and Bacteroidetes. The milk microbiome mainly comprised Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes. Alistipes spp., Ruminococcaceae UCG-10 group, Prevotellaceae UCG-001 group, and Bacteroides spp., involved in cellulose and hemicellulose degradation, enhancement of energy metabolism, and peptide breakdown, showed increment in the rectum microbiome with Aloe supplementation. The rectum microbiome in the Aloe group exhibited a significant increase in the Firmicutes to Bacteroidetes ratio and alpha-diversity at seven days after dry-off period. Beta-diversity showed a significant separation between treatments for the rectum and milk microbiomes. Aloe supplementation seemed to enrich milk microbial composition, whereas the Sealant group showed greater diversity compared to the Control group, albeit this included an increase in microorganisms frequently associated with mastitis. Conclusions Aloe arborescens administration during the dry-off period did not demonstrate any observable impact on the microbial composition of the rumen, a finding further supported by volatilome analysis. Instead, the oral Aloe supplementation at dry-off appears to significantly influence the composition of the dairy cow rectum and milk microbiomes in the following lactation.

Table olives, a vital category among fermented vegetables, come in treated and natural forms. Treated green olives require an alkaline treatment before brining to start fermentation. This study investigates the fermentation of unsalted Moroccan Picholine green olives using a method that includes heat shock treatment, inoculation with Lactiplantibacillus plantarum S61, and orange peel juice. The findings indicate that a heat shock at 70°C significantly reduces enterobacteria and enhances acidification, dropping pH levels from 5.0 to 3.6. In contrast, non-heat-treated olives showed only a slight pH decrease. The method also exhibited high antimicrobial activity, with substantial inhibition of E. coli, P. aeruginosa, L. monocytogenes, and S. aureus. Antifungal activity was noted, with significant inhibition of various strains. These results suggest that combining heat shock, L. plantarum S61, and orange peel juice can significantly improve the fermentation process of green olives, enhancing safety and quality.

Objective The aim was to study the influence of cooling milk at 9°C at the farm versus keeping it at 20°C on Parmigiano Reggiano cheese lipolysis. Methods A total of six cheesemaking trials (3 in winter and 3 in summer) were performed. In each trial, milk was divided continuously into two identical aliquots, one of which was kept at 9°C (MC9) and the other at 20°C (MC20). For each trial and milk temperature, vat milk (V-milk) and the resulting 21 month ripened cheese were analysed. Results Fat and dry matter and fat/casein ratio were lower in MC9 V-milk (p≤0.05) than in MC20. Total bacteria, mesophilic lactic acid and psychrotrophic and lipolytic bacteria showed significant differences (p≤0.05) between the two V-milks. Regarding cheese, fat content resulted lower and crude protein higher (p≤0.05) both in outer (OZ) and in inner zone (IZ) of the MC9 cheese wheels. Concerning total fatty acids, the MC9 OZ had a lower concentration of butyric, capric (p≤0.05) and medium chain fatty acids (p≤0.05), while the MC9 IZ had lower content of butyric (p≤0.05), caproic (p≤0.01) and short chain fatty acids (p≤0.05). The levels of short chain and medium chain free fatty acids (p≤0.05) were lower and that of long chain fatty acids (p≤0.05) was higher in MC9 OZ cheese. The principal component analysis of total and free fatty acids resulted in a clear separation among samples by seasons, whereas slight differences were observed between the two different milk temperatures. Conclusion Storing milk at 9°C at the herd affects the chemical composition of Parmigiano Reggiano, with repercussion on lipolysis. However, the changes are not very relevant, and since the cheese can present a high variability among the different cheese factories, such changes should be considered within the “normal variations” of Parmigiano Reggiano chemical characteristics.

In recent years, fermented foods have attracted increasing attention due to their important role in the human diet, since they supply beneficial health effects, providing important sources of nutrients. In this respect, a comprehensive characterization of the metabolite content in fermented foods is required to achieve a complete vision of physiological, microbiological, and functional traits. In the present preliminary study, the NMR-based metabolomic approach combined with chemometrics has been applied, for the first time, to investigate the metabolite content of Phaseolus vulgaris flour fermented by different lactic acid bacteria (LAB) and yeasts. A differentiation of microorganisms (LAB and yeasts), LAB metabolism (homo- and heterofermentative hexose fermentation), LAB genus (Lactobacillus, Leuconostoc, and Pediococcus), and novel genera (Lacticaseibacillus, Lactiplantibacillus, and Lentilactobacillus) was achieved. Moreover, our findings showed an increase of free amino acids and bioactive molecules, such as GABA, and a degradation of anti-nutritional compounds, such as raffinose and stachyose, confirming the beneficial effects of fermentation processes and the potential use of fermented flours in the production of healthy baking foods. Finally, among all microorganisms considered, the Lactiplantibacillus plantarum species was found to be the most effective in fermenting bean flour, as a larger amount of free amino acids were assessed in their analysis, denoting more intensive proteolytic activity.

Abstract Historic Rebel (HR) cheese is an Italian heritage cheese, produced from raw milk during the summer grazing period in the Alps. The aim of this work was (i) to characterize the cheese microbiota, by 16S rRNA gene amplicons sequencing, and the volatile and non-volatile lipophilic fraction, by Gas Chromatography and Dynamic Headspace Extraction-Gas Chromatography-Mass Spectrometry, and (ii) to evaluate their respective associations. HR cheese was dominated by Firmicutes phylum (99% of the entire abundance). The core microbiota was formed by Streptococcus, Lactobacillus, Lactococcus, Leuconostoc and Pediococcus genera together representing 87.2–99.6% of the total abundance. The polyunsaturated fatty acids composition showed a high PUFA n-3, PUFA n-6 and CLA content, two fold higher than typical plain cheeses, positively correlated with pasture altitude. A complex volatilome was detected, dominated in terms of abundance by ketones, fatty acids and alcohols. Total terpene levels increased at higher altitudes, being the main terpenes compounds α-pinene, camphene and β-pinene. The HR cheese showed a great diversity of bacterial taxa and lipophilic fractions among producers, despite belonging to a small alpine area, revealing a scarce cheese standardization and a chemical fingerprint of a typical mountain cheese produced during the grazing period. A deeper knowledge of the variability of HR cheese due to its composition in microbial community and volatile compounds will be appreciated, in particular, by elite consumers looking for niche products, adding economic value to farming in these alpine areas.

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 posture, location and social context) of 24 Alpine goats subjected to two different straw treatments (Regular: 2 kg/straw/head + 1 kg straw/head/day, n = 12; Scarce: 1 kg straw/head + 0.5 kg straw/head/day, n = 12) was observed from video recordings using a scan sampling method at 15-min scan intervals in March 2017. Moreover, straw and milk samples were submitted to microbiological analysis for the detection of total bacteria (SPC) and Enterobacteriaceae count. No significant differences in the duration of lying were observed between treatments, except at the beginning of the experimental period (Day 1: Regular: 26.0%; Scarce: 20.3% of scans; p

Among factors that can affect calf health, microbial quality of the pen air is poorly studied. In 25 Italian dairy farms, data concerning air quality in the calf pens, hygiene of pens and equipment, microclimatic conditions, calf health and management were collected during the winter season (January-March 2020 and December-March 2021). The average air Standard Plate Count (SPC) of 85 pens was 4.51 (SD = 0.52) log10 cfu/m3 whereas the average air ammonia concentration was 0.66 (SD = 0.53) ppm. Positive correlations were found between average Temperature Humidity Index (THI) in the pen and air SPC, night maximum THI and air SPC and between SPC and yeast, mould and ammonia concentration in the pen air. The concentrations of E. coli, Enterobacteriaceae and yeasts in the pen air were higher and calf cough increased as the renewal interval of bedding material became longer. High bedding dry matter and low THI were associated with low air SPC, good calf health scores and low mortality. Maintaining low bedding humidity and controlling microclimatic conditions can contribute to enhancing air microbiological quality in the pen and reduce calf diseases and mortality.

Milk microbiota represents a key point in raw milk cheese production and contributes to the development of typical flavor and texture for each type of cheese. The aim of the present study was to evaluate the influence of chlorine products usage for cleaning and sanitizing the milking equipment on (i) raw milk microbiota; (ii) the deriving whey-starter microbiota; and (iii) Trentingrana Protected Designation of Origin (PDO) cheese microbiota and volatilome. Milk samples from three farms affiliated to a Trentingrana PDO cheese factory were collected three times per week during a 6-weeks period in which a sodium hypochlorite detergent (period C) was used and during a subsequent 6-weeks period of non-chlorine detergent usage (period NC). Samples were subjected to microbiological [Standard Plate Count; coliforms; coagulase-positive staphylococci; and lactic acid bacteria (LAB)] and metagenomic analysis (amplification of V3-V4 regions of 16S rRNA gene performed on Illumina MiSeq platform). In addition, cheese volatilome was determined by SPME-GC-MS. In the transition from period C to period NC, higher SPC and LAB counts in milk were recorded. Milk metagenomic analysis showed a peculiar distinctive microbiota composition for the three farms during the whole experimental period. Moreover, differences were highlighted comparing C and NC periods in each farm. A difference in microbial population related to chlorine usage in bulk milk and vat samples was evidenced. Moreover, chlorine utilization at farm level was found to affect the whey-starter population: the usually predominant Lactobacillus helveticus was significantly reduced during NC period, whereas Lactobacillus delbrueckii had the exact opposite trend. Alpha- and beta-diversity revealed a separation between the two treatment periods with a higher presence of L. helveticus, L. delbrueckii, and Streptococcus thermophilus in cheese samples after NC detergent period. Cheese volatilome analysis showed a slight decrease in lipolysis during C period in the inner part of the cheese wheel. Although preliminary, these results suggest a profound influence on milk and cheese microbiota, as well as on raw milk cheese production and quality, due to the use of chlorine. However, further studies will be needed to better understand the complex relationship between chlorine and microbiota along all the cheese production steps.

Four LAB strains belonging to the Lactobacillus and Leuconostoc genera were investigated in experimental bread-making in comparison with Saccharomyces cerevisiae (SC). The derived breads were evaluated for microbiological and technological characteristics. Functional features associated to degradation of starch, release of gluten exorphins were investigated ante and post in vitro gastrointestinal digestion. Digestates were also assessed for rapidly digestible starch (RDS) through Englyst’s method and for secretion of the satiety hormone PYY when administered to an in vitro human intestinal cell co-culture. A clear separation emerged between LAB and SC samples, being the highest difference affected by TTA and reduction of phytate. The SC sample was also distinguishable by its highest levels of simple sugars, in particular glucose, post in vitro digestion and PYY secretion. Breads from LAB strains differed with regard to volume and moisture, TTA, phytate reduction, exorphins release, total sugars and glucose contents post in vitro digestion.

Food spoilage is a serious issue dramatically impacting the worldwide need to counteract food insecurity. Despite the very expensive application of low temperatures, the proper conservation of fresh dairy products is continuously threatened at different stages of production and commercialization by psychrotrophic populations mainly belonging to the Pseudomonas genus. These bacteria cause discolouration, loss of structure, and off-flavours, with fatal implications on the quality and shelf-life of products. While the effects of pseudomonad decay have been widely reported, the mechanisms responsible for the activation and regulation of spoilage pathways are still poorly explored. Recently, molecule signals and regulators involved in quorum sensing (QS), such as homoserine lactones, the luxR/luxI system, hdtS, and psoR, have been detected in spoiled products and bacterial spoiler species; this evidence suggests the role of bacterial cross talk in dairy spoilage and paves the way towards the search for novel preservation strategies based on QS inhibition. The aim of this review was to investigate the advancements achieved by the application of omic approaches in deciphering the molecular mechanisms controlled by QS systems in pseudomonads, by focusing on the regulators and metabolic pathways responsible for spoilage of fresh dairy products. In addition, due the ability of pseudomonads to quickly spread in the environment as biofilm communities, which may also include pathogenic and multidrug-resistant (MDR) species, the risk derived from the gaps in clearly defined and regulated sanitization actions is underlined.

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.

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.

Abstract Sustainable exploitation of agro-industrial by-products has attracted great interest in cereal bran valorization. In this research, a polyphasic approach has been carried out to characterize maize bran at microbiological and chemical level during a sourdough like fermentation process, in order to enhance its technological and nutritional properties. Autochthonous microbiota was isolated at different refreshment steps and subjected to identification and molecular characterization. Fermentation was characterized by a rapid increase in lactic acid bacteria and yeasts, with a co-dominance, at the initial stage, of Weissella spp., Pediococcus spp. and Wickerhamomyces anomalus. At the end of the fermentation, a natural selection was produced, with the prevalence of Lactobacillus plantarum, Lactobacillus brevis and Kazachstania unispora. This is the first time that a specific association between LAB and yeasts is reported, during the maize bran fermentation process. Enzymatic activities related to this microbial consortium promoted a “destructuration” of the fiber fraction, an increase in soluble dietary fiber and a reduction of phytic acid content. Our data also evidenced a noticeable increment in ferulic acid. The results obtained indicate that fermentation processes represent an efficient biotechnological approach to increase nutritional and functional potential of maize bran. Moreover, the characterization of microbiota involved in natural fermentation process will allow the selection of specific biotypes, with appropriate metabolic and enzymatic activities, to conduct “tailored” fermentation processes and improve brans or whole-meal flours from both nutritional and technological points of view.

Plants develop in a microbe-rich environment and must interact with a plethora of microorganisms, both pathogenic and beneficial. Indeed, such is the case of Pseudomonas, and its model organisms P. fluorescens and P. syringae, a bacterial genus that has received particular attention because of its beneficial effect on plants and its pathogenic strains. The present study aims to compare plant-beneficial and pathogenic strains belonging to the P. syringae species to get new insights into the distinction between the two types of plant–microbe interactions. In assays carried out under greenhouse conditions, P. syringae pv. syringae strain 260-02 was shown to promote plant-growth and to exert biocontrol of P. syringae pv. tomato strain DC3000, against the Botrytis cinerea fungus and the Cymbidium Ringspot Virus. This P. syringae strain also had a distinct volatile emission profile, as well as a different plant-colonization pattern, visualized by confocal microscopy and gfp labeled strains, compared to strain DC3000. Despite the different behavior, the P. syringae strain 260-02 showed great similarity to pathogenic strains at a genomic level. However, genome analyses highlighted a few differences that form the basis for the following hypotheses regarding strain 260-02. P. syringae strain 260-02: (i) possesses non-functional virulence genes, like the mangotoxin-producing operon Mbo; (ii) has different regulation pathways, suggested by the difference in the autoinducer system and the lack of a virulence activator gene; (iii) has genes encoding DNA methylases different from those found in other P. syringae strains, suggested by the presence of horizontal-gene-transfer-obtained methylases that could affect gene expression.

The proteolytic traits of the psychrotrophic strains Pseudomonas poae LP5, Pseudomonas fluorescens LPF3, Chryseobacterium joostei LPR1, Pseudomonas fulva PS1, Citrobacter freundii PS37, Hafnia alvei PS46, and Serratia marcescens PS92 were initially investigated by phenotypic and genotypic approaches. Six strains elicited extracellular proteolytic activity, and five expressed the thermostable AprX or (likely) Ser1 enzymes. Then, the strains were inoculated (104 CFU/mL) in microfiltered pasteurized milk and kept at 4 °C for five days. All of the strains reached 108 CFU/mL at the end of storage and five produced thermostable extracellular proteolytic enzymes. The freshly inoculated samples and the corresponding samples at 108 CFU/mL were batch-sterilized (131 °C, 30 s) and kept at 45 °C up to 100 days. The former samples did not gel until the end of incubation, whereas the latter, containing P. poae, P. fluorescens, C. joostei, C. freundii, and S. marcescens, gelled within a few days of incubation. The thermostable proteolytic activity of strains affected the peptidomic profile, and specific proteolyzed zones of β-CN were recognized in the gelled samples. Overall, the results confirm some proteolytic traits of psychrotrophic Pseudomonas spp. strains and provide additional insights on the proteolytic activity of psychrotrophic bacteria potentially responsible for sterilized milk destabilization.

Ready-to-eat salads are very perishable with quality losses within 6–7 days, and the extension of their shelf life is still a challenge. In this work, an atmospheric pressure plasma jet (APPJ) was applied for the surface decontamination of fresh-cut lettuce baby leaves. The APPJ antimicrobial efficiency on the natural microbiota and its impact on some physicochemical attributes of lettuce were evaluated as a function of the treatment duration (0–30 s). Then, the influence of plasma treatment on the salad shelf life was studied, following the growth of aerobic mesophilic bacteria in both untreated and plasma-treated samples during 9 days of storage at 4 °C, together with the plasma-induced changes in physicochemical parameters of lettuce leaves. The APPJ induced a fast (15 s) microbial decontamination (1.3 log10 CFU/g) of the salad surface. Exposure time and salad-plasma plume distance were the parameters that substantially affected the microbial inactivation. APPJ treatment retarded bacterial growth during the refrigerated storage, as plasma-treated samples were noticeably less contaminated than the non-treated ones in the first 3–4 days. No significant effect were observed on electrolyte leakage, pH, and dry matter content in both the set up phase and the shelf life study.

The natural load of lactic acid bacteria (LAB) in milk is the basis of the production of raw milk cheeses, such as Grana Padano PDO. In the last decades, improvements in livestock hygiene management resulted in bulk cow milk with less than 20,000 colony forming units (CFU) of bacterial count, unable to ensure a sufficient supply of LAB, with a negative impact on cheese quality. This study investigated the relations between farm management practices and prevalence of different groups of bacteria in cow milk. Sixty-two intensive dairy farms located in Lombardy (Italy) where involved, most of them destined as milk for the production of Grana Padano. Season had no significant effect on the content of most of the bacterial groups, except for coliforms. A strong relation among standard plate count (SPC) and other bacterial groups was evidenced. Cluster analysis showed that the most productive farms applied a complete milking routine and produced milk with the lowest value of SPC, the lowest count of the other bacteria, including LAB, but the highest LAB/SPC. The study suggests that complexity of farming practices can affect the microbial population of milk.

The aim of the study is to develop a scoring system for dairy farms in order to give specific information about the product and production process of milk. The scoring system, based on a multi-dimensional approach, was developed on the basis of data collected in 29 Italian dairy farms and included different aspects. For the evaluation of animal welfare, a selection of indicators set up in the European Project® assessment protocol for cattle 2009 was used. Environmental sustainability of milk production was assessed by a cradle-to-farm-gate Life Cycle Assessment. Laboratory analyses were carried out on bulk tank milk to evaluate microbiological, nutritional and nutraceutical quality. Nineteen variables were selected and retained to define six quality aspects: animal welfare, environmental and economic sustainability of farms, microbiological, nutritional and nutraceutical quality of milk. Each farm was visited twice; each visit received, for each variable, a score between 1 and 3 based on the frequency distribution of that variable in the farm sample. The relation among farm characteristics and quality aspects showed the importance to maximise dairy efficiency to improve environmental and economic sustainability of the farms and the inclusion of hay in dairy cows’ ration to enhance the nutraceutical and nutritional quality of milk. The proposed multi-dimensional scoring system is a practical tool: for the farmer, to support decisions for improving the quality of the product and the productive process; for the dairy company, as a value-added opportunity; for the consumer, who receives detailed information about nutritional characteristics and production system of dairy products.

Although camelina [Camelina sativa (L.) Crantz] is a good source of protein, antioxidants, and polyunsaturated fatty acids, its antinutritional compounds limit its use in animal feeding. The aim of this study was to verify the effect of feeding laying hens with up to 20% of camelina cake from a breeding line containing a low level of glucosinolates on performance, welfare, and eggshell quality. Two hundred and forty Hy-Line® hens from 18 to 51 weeks of age were divided into three treatments: control (C), camelina cake 10% (CAM10), and camelina cake 20% (CAM20). Egg number was recorded daily, while egg weight, feed consumption, and mortality were recorded weekly. At 24 and 43 weeks of hen age, shell resistance to fracture was measured. Our results demonstrate no detrimental effects for CAM10 and CAM20 diets on feed intake, growth performance, and welfare. No difference in egg production was detected among the diets. The significant (p < 0.05) interaction of diet and age factors suggest that the addition of camelina cake, up to 20%, likely protects the eggshell of older hens. Our findings confirm that camelina cake might be an alternative and sustainable protein source for hens.

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 < 0.001), while SFA showed higher values in S (68.85 vs 63.41 and 61.68 in P and A, respectively; p < 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 < 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.

Bacteria of the Paenibacillus genus are becoming important in many fields of science, including agriculture, for their positive effects on the health of plants. However, there are little information available on this genus compared to other bacteria (such as Bacillus or Pseudomonas), especially when considering genomic information. Sequencing the genomes of plant-beneficial bacteria is a crucial step to identify the genetic elements underlying the adaptation to life inside a plant host and, in particular, which of these features determine the differences between a helpful microorganism and a pathogenic one. In this study, we have characterized the genome of Paenibacillus pasadenensis, strain R16, recently investigated for its antifungal activities and plant-associated features. An hybrid assembly approach was used integrating the very precise reads obtained by Illumina technology and long fragments acquired with Oxford Nanopore Technology (ONT) sequencing. De novo genome assembly based solely on Illumina reads generated a relatively fragmented assembly of 5.72 Mbp in 99 ungapped sequences with an N50 length of 544 Kbp; hybrid assembly, integrating Illumina and ONT reads, improved the assembly quality, generating a genome of 5.75 Mbp, organized in 6 contigs with an N50 length of 3.4 Mbp. Annotation of the latter genome identified 4987 coding sequences, of which 1610 are hypothetical proteins. Enrichment analysis identified pathways of particular interest for the endophyte biology, including the chitin-utilization pathway and the incomplete siderophore pathway which hints at siderophore parasitism. In addition the analysis led to the identification of genes for the production of terpenes, as for example farnesol, that was hypothesized as the main antifungal molecule produced by the strain. The functional analysis on the genome confirmed several plant-associated, plant-growth promotion, and biocontrol traits of strain R16, thus adding insights in the genetic bases of these complex features, and of the Paenibacillus genus in general.

Hen egg-white lysozyme (LSZ) is currently used in the food industry to limit the proliferation of lactic acid bacteria spoilage in the production of wine and beer, and to inhibit butyric acid fermentation in hard and extra hard cheeses (late blowing) caused by the outgrowth of clostridial spores. The aim of this work was to evaluate how the enzyme activity in commercial preparations correlates to the enzyme concentration and can be affected by the presence of process-related impurities. Different analytical approaches, including turbidimetric assay, SDS-PAGE and HPLC were used to analyse 17 commercial preparations of LSZ marketed in different countries. The HPLC method adopted by ISO allowed the true LSZ concentration to be determined with accuracy. The turbidimetric assay was the most suitable method to evaluate LSZ activity, whereas SDS-PAGE allowed the presence of other egg proteins, which are potential allergens, to be detected. The analytical results showed that the purity of commercially available enzyme preparations can vary significantly, and evidenced the effectiveness of combining different analytical approaches in this type of control.

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

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.