Thymol in the intestinal tract of broiler chickens after sustained administration of thyme essential oil in feed.

Plant compounds occurring in phytogenic feed additives are involved in different pharmacological activities in the animal organism. Since the digestive tract acts as a first line of defence against foreign compounds, it is necessary to outline its response to dietary supplementation with bioactive plant components. Little information is available on the bioactivity of thymol as the main bioactive compound of Thymus vulgaris L. essential oil (TEO). The main objective of the present study was to provide a detailed view of the concentrations of thymol in plasma and the content of individual intestinal segments (duodenum, jejunum, ileum, caecum and colon) of broiler chickens after 4 weeks of dietary supplementation with different TEO concentrations. 32 one-day old Ross 308 hybrid broilers were randomly divided into four dietary treatment groups (0.00%, 0.01%, 0.05%, 0.1% w/w of TEO in the diet). Thymol concentrations in the duodenal chyme presented around 7% on average from the thymol amount administered in the feed. A significantly increased thymol amount was observed after 0.1% TEO addition to the diet compared with 0.01% TEO enrichment in the duodenal wall and gut content of jejunum, ileum, caecum and colon (p < 0.05). Thymol levels in the colon were significantly higher than in the ileum and about 1.7 times higher on average than those in the caecum. Significant coefficient of correlation was observed between thymol concentrations in plasma and feed, gut content of all intestinal segments as well as duodenal wall. Our results point to intensive thymol absorption in the initial sections of the digestive tract. In the current study, the role of intestine in biotransformation of thymol was observed, and it would be desirable to investigate whether thymol itself or thymol metabolites are responsible for beneficial effects in intestine.

Activity of 30 different cheeses on cholesterol plasma levels and Oxidative Balance Risk Index (OBRI) in a rat model.

BACKGROUND: Cheese is considered to increase the total cholesterol levels (CH) due to the high-saturated fat content. New models are needed to measure the relationship between cholesterol and cheese. METHODS: Thirty different cheeses produced in Val Brembana, Italy ("furmai da mut", "caprino" and "stracchino"), were added to the diet of 30 groups of 4 rats. Cheeses were analyzed to differentiate the volatile organic compounds (VOCs) and the cholesterol content (Ch(f)). The body weight, CH, urine volume and oxidative balance were measured. Three new indexes in relation to CH were calculated: OI (oxidative index), PI (protective index) and OBRI (oxidative balance risk index). RESULTS: None of the cheeses increased CH. Some of the "furmai de mut" were significantly decreasing CH and improved the oxidative balance. Ch(f) was not affecting the CH levels in plasma. In terms of VOCs, the acetic acid content was correlated (p < 0.05) with the CH reduction and PI improvement. OBRI was reduced mainly in the "stracchino group". CONCLUSIONS: The model shows that some cheese can reduce significantly CH levels and improve the antioxidant capacity.

High biodiversity in a limited mountain area revealed in the traditional production of Historic Rebel cheese by an integrated microbiota-lipidomic approach.

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 ß-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.

Effect of Linseeds and Hemp Seeds on Milk Production, Energy and Nitrogen Balance, and Methane Emissions in the Dairy Goat.

The effect of whole linseeds or hemp seeds on milk production, energy and nitrogen balance, and methane emission was studied in 12 Alpine goats using respiration chambers. Diets tested were a control diet (C) and two diets supplemented with whole linseeds (L) or hemp seeds (H) at 9.3% on a dry matter (DM) basis. DM intake was similar among treatments, whereas DM and organic matter digestibility were lower for L compared to C. Milk yield (2.30 kg/d on average) and rumen fermentation profile were not affected by treatments. Treatment also did not affect the milk composition, with the exception of fat, which was higher in H and L compared to C (4.21, 3.94, and 3.20%, respectively). Oilseed supplementation caused a reduction in the concentration of de novo fatty acids (FA) (41.1, 48.8, and 64.1% of FA, for L, H, and C, respectively). Moreover, L and H diets reduced the sum of saturated FA, and increased monounsaturated FA, whereas only the L diet increased the concentration of polyunsaturated FA. Regarding methane production, and nitrogen and energy balances, no differences were registered among the diets. Our research indicates that including whole linseeds and hemp seeds in the dairy goat diet is an effective strategy for increasing milk fat content and positively modifying the milk FA composition, without a change in nitrogen and energy balances, but also without a reduction in enteric methane emission.

Characterization of Lysinibacillus fusiformis strain S4C11: In vitro, in planta, and in silico analyses reveal a plant-beneficial microbe.

Despite sharing many of the traits that have allowed the genus Bacillus to gain recognition for its agricultural relevance, the genus Lysinibacillus is not as well-known and studied. The present study employs in vitro, in vivo, in planta, and in silico approaches to characterize Lysinibacillus fusiformis strain S4C11, isolated from the roots of an apple tree in northern Italy. The in vitro and in vivo assays demonstrated that strain S4C11 possesses an antifungal activity against different fungal pathogens, and is capable of interfering with the germination of Botrytis cinerea conidia, as well as of inhibiting its growth through the production of volatile organic molecules. In planta assays showed that the strain possesses the ability to promote plant growth, that is not host-specific, both in controlled conditions and in a commercial nursery. Biocontrol assays carried out against phytopathogenic viruses gave contrasting results, suggesting that the strain does not activate the host's defense pathways. The in silico analyses were carried out by sequencing the genome of the strain through an innovative approach that combines Illumina and High-Definition Mapping methods, allowing the reconstruction of a main chromosome and two plasmids from strain S4C11. The analysis of the genes encoded by the genome contributed to the characterization of the strain, detecting genes related to the biocontrol effect detected in the experimental trials.

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 < 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.

Effect of Different Percentage of Camelina sativa Cake in Laying Hens Diet: Performance, Welfare, and Eggshell Quality.

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.

Raw Milk Microbiota Modifications as Affected by Chlorine Usage for Cleaning Procedures: The Trentingrana PDO Case.

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.

Not Just a Pathogen? Description of a Plant-Beneficial Pseudomonas syringae Strain.

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.

Terpenes and fatty acid profiles of milk fat and "Bitto" cheese as affected by transhumance of cows on different mountain pastures.

The evolution of fatty acid (FA) and terpenoid profiles was studied in milk (n=20) and "Bitto" (n=3), a protected designation of origin cheese produced in a restricted Italian alpine area. Milk came from 25 Italian Brown cows successively grazing pastures at 1400, 2100 and 2200m during transhumance in June-September 2006. The fat matter was analyzed for FAs and terpenes by means of gas chromatography and purge & trap/gas chromatography-mass spectrometry, respectively. FA composition of milk fat varied significantly (p<0.0001) in relation to contents of conjugated linoleic acid (CLA), stearic, linoleic and trans-vaccenic acids. Similar monoterpene profiles characterized milk fat from cows grazing the different pastures and the highest amount of terpenes was measured in milk coming from cows grazing at 1400m. High levels of δ3-carene in milk fat were likely related to the important presence of Ligusticum mutellina in the pasture. Only negligible amounts of sesquiterpenes were detected in milk fat whereas they were the most abundant class in fodder. Both FA and terpene profiles of ripened (70 days) cheeses resembled those of the original milks. Overall, results confirm the influence of the botanical composition of mountain pastures both in enhancing the ruminal synthesis of CLA and in modifying the FA and terpenoid profiles of milk and "Bitto" cheese. Nevertheless, neither the FA nor the terpenoid profiles revealed here can be considered as "unique" to "Bitto" cheese and, for this reason, they can hardly be assumed to be biomarkers for defining a specific relationship among grazing area, milk and "Bitto" cheese. They better represent the chemical fingerprint of the cow feeding, adopted in mountain areas.

Competition assays revealed Paenibacillus pasadenensis strain R16 as a novel antifungal agent.

The development of new sustainable containment strategies of plant diseases is very important to guarantee food security while reducing the environmental impact of agriculture. Research of new biocontrol agents is a long and difficult process that involves several steps that start from the identification of possible candidates which, for example, show antibiotic activities, and ends with in field, large scale trials. In this study, the plant growth promoting potential and antifungal effect exerted by a novel, putative candidate biocontrol agent, strain R16, identified as Paenibacillus pasadenensis by sequence analysis of 16S rRNA and rpoB genes, against three important plant pathogenic fungi (Botrytis cinerea, Fusarium verticillioides, and Phomopsis viticola), were assessed. Biochemical assays to determine plant growth promoting potential gave negative results for siderophore production and phosphate solubilization, and positive results for ACC-deamination and IAA production. Further biochemical assays for endophytic lifestyle and antifungal activity gave positive results for catalase and chitinase activity, respectively. In vitro antagonism assays showed that strain R16 is effective against B. cinerea, reducing mycelial growth both in dual-culture and through volatile substances, characterized to be mostly composed by farnesol, and inhibiting conidia germination. Good antagonistic potential was also observed in vitro towards P. viticola, but not towards F. verticillioides. Moreover, in vivo assays confirmed the strain R16 activity reduced the infection rate on B. cinerea-inoculated berries. The obtained results firstly proved that P. pasadenesis strain R16 is a putative plant growth promoter and effective against phytopathogenic fungi. Further studies will be needed to investigate the possible application of this strain as a biocontrol agent.

Variability in Seed Traits in a Collection of Cannabis sativa L. Genotypes.

The seed of Cannabis sativa L. is an expanding source of proteins and oil for both humans and animals. In this study, the proximate composition of a collection of hemp cultivars and accessions of different geographical origins grown under the same conditions for 1 year was analyzed in order to identify potential accessions to improve hemp cultivars. Fatty acids, tocopherols, and antinutritional components, as well as concentrations of crude protein and oil were quantified. The seed oil concentrations varied between 285 and 360 g kg(-1) dry seed (DS), while crude protein ranged between 316 and 356 g kg(-1) dry matter (DM). The seed oil was mainly composed of unsaturated fatty acids and, as expected, the dominant fatty acids were linoleic and α-linolenic acid. A high variability among the cultivars and accessions was also detected for polyphenolic content which ranged from 5.88 to 10.63 g kg(-1) DM, cv. Felina was the richest, whereas cv. Finola had the lowest polyphenolic content. Regarding antinutritional compounds in seed, a high variability was detected among all genotypes analyzed and phytic acid was particularly abundant (ranging between 43 and 75 g kg(-1) DM). In conclusion, our results reveal noticeable differences among hemp seed genotypes for antinutritional components, oil and protein content. Collectively, this study suggests that the hemp seed is an interesting product in terms of protein, oil and antioxidant molecules but a reduction of phytic acid would be desirable for both humans and monogastric animals. The high variability detected among the different genotypes indicates that an improvement of hemp seed might be possible by conventional and/or molecular breeding.