The Effect of Dietary Supplementation with Probiotic and Postbiotic Yeast Products on Ewes Milk Performance and Immune Oxidative Status.

The administration of yeast products as feed additives has been proven to beneficially affect animal productivity through energy, oxidative, and immune status improvement. This study evaluated a combination of Saccharomyces cerevisiae live yeast (LY) with yeast postbiotics (rich in mannan-oligosaccharides (MOS) and beta-glucans) and selenium (Se)-enriched yeast on ewes' milk performance and milk quality, energy and oxidative status, and gene expression related to their immune system during the peripartum period. Ewes were fed a basal diet (BD; F:C = 58:42 prepartum and 41:59 postpartum) including inorganic Se (CON; n = 27), the BD supplemented with a LY product, and inorganic Se (AC; n = 29), as well as the combination of the LY, a product of yeast fraction rich in MOS and beta-glucans, and organic-Se-enriched yeast (ACMAN; n = 26) from 6 weeks prepartum to 6 weeks postpartum. The ß-hydroxybutyric acid concentration in the blood of AC and ACMAN ewes was lower (compared to the CON) in both pre- and postpartum periods (p < 0.010). Postpartum, milk yield was increased in the AC and ACMAN Lacaune ewes (p = 0.001). In addition, the activity of superoxide dismutase (p = 0.037) and total antioxidant capacity (p = 0.034) measured via the 2,2-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) method was increased in the blood plasma of the ACMAN postpartum. Higher ABTS values were also found (p = 0.021), while protein carbonyls were reduced (p = 0.023) in the milk of the treated groups. The relative transcript levels of CCL5 and IL6 were downregulated in the monocytes (p = 0.007 and p = 0.026 respectively), and those of NFKB were downregulated in the neutrophils of the ACMAN-fed ewes postpartum (p = 0.020). The dietary supplementation of ewes with yeast postbiotics rich in MOS and beta-glucans, and organic Se, improved energy status, milk yield and some milk constituents, and oxidative status, with simultaneous suppression of mRNA levels of proinflammatory genes during the peripartum period.

Dietary Supplementation of a Live Yeast Product on Dairy Sheep Milk Performance, Oxidative and Immune Status in Peripartum Period.

This study evaluated the dietary administration of Saccharomyces cerevisiae live yeast on milk performance and composition, oxidative status of both blood plasma and milk, and gene expression related to the immune system of lactating ewes during the peripartum period. Chios ewes were fed either a basal diet (BD) (Control, n = 51) or the BD supplemented with 2 g of a live yeast product/animal (ActiSaf, n = 53) from 6 weeks prepartum to 6 weeks postpartum. Fatty acid profile, oxidative, and immune status were assessed in eight ewes per treatment at 3 and 6 weeks postpartum. The ß-hydroxybutyric acid concentration in blood of ActiSaf fed ewes was significantly lower in both pre- and postpartum periods. A numerical increase was found for the milk yield, fat 6% corrected milk (Fat corrected milk (FCM6%)), and energy corrected milk yield (ECM) in ActiSaf fed ewes, while daily milk fat production tended to increase. The proportions of C15:0, C16:1, C18:2n6t, and C18:3n3 fatty acids were increased in milk of ActiSaf fed ewes, while C18:0 was decreased. Glutathione reductase in blood plasma was increased (p = 0.004) in ActiSaf fed ewes, while total antioxidant capacity measured by 2,2'-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) method was decreased (p < 0.001). Higher ABTS values were found in the milk of the treated group. The relative transcript levels of CCL5, CXCL16, and IL8 were suppressed, while that of IL1B tended to decrease (p = 0.087) in monocytes of ActiSaf fed ewes. In conclusion, the dietary supplementation of ewes with S. cerevisiae, improved the energy utilization and tended to enhance milk performance with simultaneous suppression on mRNA levels of pro-inflammatory genes during the peripartum period.

Bioactive Compounds in Food Waste: A Review on the Transformation of Food Waste to Animal Feed.

Bioactive compounds are substances which are present in foods in small amounts and have the ability to provide health benefits. Bioactive compounds include but are not limited to long-chain polyunsaturated fatty acids, vitamins, carotenoids, peptides, and polyphenols. The aim of the present study is to review literature for potential bioactive compounds present in food waste material and discuss the transformation of food waste to animal feed under the perspective that usage of food waste, rather than disposal, may tackle food insecurity and provide health benefits. Finally, applications in poultry and swine nutrition, with emphasis on the presence of fatty acids on food waste material, are discussed.

The effects of dietary supplementation with rumen-protected amino acids on the expression of several genes involved in the immune system of dairy sheep.

Amino acids might be a tool to transform animals from a pro- to an anti-inflammatory phenotype through the downregulation of several genes (TLR-4, NF-κB, TNFa, IL-1ß, IL-2, IL-6, IL-8, CCL-5 and CXCL-16) whose expression increases during inflammation. To examine this possibility, each of sixty Chios dairy sheep was assigned to one of the following five dietary treatments: A: basal diet (control group); B: basal diet +6.0 g/head rumen-protected methionine (MetaSmart™ ); C: basal diet +5.0 g/head rumen-protected lysine (LysiGEM™ ); D: basal diet +6.0 g/head MetaSmart™  + 5.0 g/head LysiGEM™ ; and E: basal diet +12.0 g/head MetaSmart™  + 5.0 g/head LysiGEM. The results showed a significant downregulation in the expression of the TLR-4 gene in both the macrophages and the neutrophils of ewes fed rumen-protected amino acids. Significantly lower mRNA transcript accumulation was also observed for the TNFa, IL-1ß and CXCL-16 genes in the macrophages and for the IL-1ß gene in the neutrophils of ewes supplemented with amino acids. The ewes that received dietary supplementation with rumen-protected lysine alone (C) had significantly lower CCL-5 transcript levels in their macrophages than the ewes fed the other supplemented diets. Diet D enhanced the mRNA expression of the IL-2 gene in ewe neutrophils. Negative correlations were found between: a. TLR-4, TNFa, IL-1ß and CXCL-16 gene expression in macrophages and the milk fat and total solids content; b. CCL-5 gene expression in neutrophils and the milk yield and FCM(6%) ; and c. CXCL-16 gene expression and the milk protein content. Moreover, positive correlations were found between the BHBA concentration and the expression of the TLR-4 and CXCL-16 genes in macrophages. In conclusion, the rumen-protected amino acids improved sheep metabolism (as indicated by reduced blood BHBA and urea concentrations), milk chemical composition and immune system function.

The effect of long term under- and over-feeding on the expression of six major milk protein genes in the mammary tissue of sheep.

Milk protein synthesis in the mammary gland involves expression of six major milk protein genes whose nutritional regulation remains poorly defined. In this study, the effect of long term under- and over-feeding on the expression of αs1-casein: CSN1S1, αs2-casein: CSN1S2, ß-casein: CSN2, κ-casein: CSN3, α-lactalbumin: LALBA and ß-lactoglobulin: BLG gene in sheep mammary tissue (MT) was examined. Twenty-four lactating dairy sheep, at 90-98 d in milk, were divided into three groups and fed the same ration, for 60 d, in quantities which met 70% (underfeeding), 100% (control) and 130% (overfeeding) of their energy and crude protein requirements. The results showed a significant reduction on mRNA of CSN1S1, CSN1S2, CSN2 and BLG gene in the MT of underfed sheep compared with the overfed ones and a significant reduction in CSN3 and LALBA gene expression compared with the respective control animals. Significant positive correlations were observed between the mRNA levels of milk proteins' genes with the milk protein yield and milk yield respectively. In conclusion, the feeding level and consequently the nutrients availability, affected the milk protein yield and milk volume by altering the CSN1S1, CSN1S2, CSN2, CSN3, LALBA and BLG gene expression involved in their metabolic pathways.

The effect of long term under- and over-feeding on the expression of genes related to glucose metabolism in mammary tissue of sheep.

Glucose utilisation for lactose synthesis in the mammary gland involves expression of a large number of genes whose nutritional regulation remains poorly defined. In this study, the effect of long term under- and over-feeding on the expression of genes [glucose transporter 1: GLUT1, glucose transporter 3: GLUT3, Sodium glucose contransporter 1: SGLT1, two isoforms of ß- (1,4) galactosyltransferase: ß- (1,4) GAT1, ß- (1,4) GAT3 and α-lactalbumin: LALBA] related to glucose metabolism in sheep mammary tissue (MT) was examined. Twenty-four lactating dairy sheep were divided into three homogenous sub-groups and fed the same ration in quantities which met 70% (underfeeding), 100% (control) and 130% (overfeeding) of their energy and crude protein requirements. The results showed a significant reduction on mRNA of GLUT1 and LALBA gene in the MT of underfed sheep, compared with the respective controls and overfed and a significant reduction on mRNA level of SGLT1 and ß- (1,4) GAT1 in the MT of underfed sheep, compared with the overfed ones. A significant increase in the GLUT3 mRNA accumulation in the MT of both under- and over- fed sheep was found. Additionally, a trend of reduction on ß- (1,4) GAT3 mRNA level in the MT of the underfed sheep, compared with the overfed, was observed. A close positive relationship was obtained between the mRNA transcripts accumulation of GLUT1, SGLT1, ß- (1,4) GAT1 and LALBA gene with the milk lactose content and milk lactose yield respectively. In conclusion, feeding level and consequently nutrient availability, may affect glucose uptake and utilisation in sheep MT by altering the GLUT1, GLUT3, SGLT1, ß- (1,4) GAT1 and LALBA gene expression involved in their metabolic pathways.

The effect of long term under- and over-feeding on the expression of genes related to lipid metabolism in mammary tissue of sheep.

Milk fatty acid (FA) synthesis by the mammary gland involves expression of a large number of genes whose nutritional regulation remains poorly defined. In this study, we examined the effect of long-term under- and over-feeding on the expression of genes (acetyl Co A carboxylase, ACC; fatty acid synthetase, FAS; lipoprotein lipase, LPL; stearoyl Co A desaturase, SCD; peroxisome proliferator activated receptor γ2, PPARγ2; sterol regulatory element binding protein-1, SREBP-1c; and hormone sensitive lipase, HSL) related to FA metabolism in sheep mammary tissue (MT). Twenty-four lactating sheep were divided into three homogenous sub-groups and fed the same ration in quantities covering 70% (underfeeding), 100% (control) and 130% (overfeeding) of their energy and crude protein requirements. The results showed a significant reduction of mRNA of ACC, FAS, LPL and SCD in the MT of underfed sheep, and a significant increase on the mRNA of LPL and SREBP-1c in the MT of overfed compared with the control respectively. In conclusion, the negative, compared to positive, energy balance in sheep down-regulates ACC, FAS, LPL, SCD, SREBP-1c and PPARγ2 expression in their MT which indicates that the decrease in nutrient availability may lead to lower rates of lipid synthesis.

Effects of high selenium and fat supplementation on growth performance and thyroid hormones concentration of broilers.

A total of 400, as hatched, broilers were used to investigate the effect of increase of selenium and energy intake on thyroid hormone metabolism, growth and liver fatty acid profile. There were 5 replicates of 4 dietary treatments namely, TA (0.289mg Se per kg diet and adequate energy content), TB (0.583mg Se per kg diet and adequate energy content), TC (0.267mg Se per kg diet and 9% increase of energy content) and TD (0.576mg Se per kg diet and 9% increase of energy content). Diets were isonitrogenous. Zinc L-selenomethionine complex was used to increase Se content and corn oil was used to increase the energy content. The experiment lasted 42 days. Broiler growth performance was not significantly affected by dietary treatments. Liver glutathione peroxidase (GPx) activity increased (P<0.05) in broilers fed high Se and energy diets compared to other ones. Whole blood GPx activity was higher in Se supplemented groups however, it was reduced by age. Thyroid hormone concentrations were unaffected by dietary treatments. A significant increase of linoleic and arachidonic acid concentration (P<0.001) was observed in the liver of broilers fed diets with moderately increased energy content and supplemented with Se compared to those fed diets with moderately increased energy content alone. In conclusion, zinc L-selenomethionine complex and moderate increase of energy content did not affect growth rate or thyroid hormone metabolism but led to increased liver fatty acid content and hepatic GPx activity.

Changes in milk and plasma fatty acid profile in response to fish and soybean oil supplementation in dairy sheep.

An effective strategy for enhancing the bioactive fatty acids (FA) in sheep milk could be dietary supplementation with a moderate level of a combination of soybean oil with fish oil (SFO) without negative effects on milk yield and its chemical composition. Thus, the objective of this study was to determine the effects of a moderate forage diet supplementation with SFO on milk chemical composition and FA profile, as well as on plasma FA. Twelve dairy sheep were assigned to two homogenous sub-groups. Treatments involved a control diet without added oil, and a diet supplemented with 23.6 g soybean oil and 4.7 g fish oil per kg dry matter (DM) of the total ration. The results showed that SFO diet had no effect on milk yield and chemical composition. In blood plasma the concentrations of trans-11 C(18:2) (VA), C(18:2n-6), C(20:5n-3) (EPA) and C(22:6n-3) (DHA) were significantly higher while those of C(14:0), C(16:0) and C(18:0) were lower in sheep fed with SFO diet compared with control. The SFO supplementation of sheep diet increased the concentrations of VA, cis-9, trans-11 C(18:2) CLA, trans-10, cis-12, C(18:2) CLA, EPA, DHA, monounsaturated FA (MUFA), polyusaturated fatty acids (PUFA) and n-3 FA and decreased those of short chain FA (SCFA), medium chain FA (MCFA), the saturated/unsaturated ratio and the atherogenicity index value in milk compared with the control. In conclussion, the SFO supplementation at the above levels in a sheep diet, with moderate forage to concentrate ratio, improved the milk FA profile from human health standpoint without negative effects on its chemical composition.

The effect of long term under- and over-feeding of sheep on milk and plasma fatty acid profiles and on insulin and leptin concentrations.

Since sheep's milk is mainly used for cheese making and milk chemical composition and fatty acids (FA) profile affect cheese yield and quality, the objective of this study was to determine the effects of different feeding levels on milk chemical composition and FA profile, as well as on plasma FA profile, and on insulin and leptin concentrations. Twenty-four sheep were assigned to three homogeneous sub-groups. Throughout the experimental period each group was fed the same diet but in quantities which met 70% (under-feeding), 100% (control) and 130% (over-feeding) of their respective energy and crude protein requirements. The results showed that the underfed sheep had higher milk fat content compared with overfed. In blood plasma the concentrations of C18:0 and C18:1 in the underfed sheep were significantly higher compared with control and overfed sheep. The concentrations of leptin and insulin were significantly higher in overfed compared with underfed sheep. Underfeeding reduced the concentrations of short chain FA (SCFA) and medium chain FA (MCFA) and increased that of C18:0 and mono unsaturated FA (MUFA) in sheep milk fat compared with controls and overfed. The concentrations of C18:0, long chain FA (LCFA) and monounsaturated FA (MUFA) in milk were significantly higher and those of SCFA, MCFA and saturated FA (SFA) significantly lower in the underfed compared with the overfed sheep. In conclusion, long term under- and over-feeding affected the sheep milk chemical composition and FA profile which consequently has an impact on milk products yield (cheese and yogurt) and quality (human health).

Differences in sheep and goats milk fatty acid profile between conventional and organic farming systems.

The objective of this study was to investigate whether there is a difference in chemical composition and particularly in fatty acid (FA) profile, with emphasis on cis-9, trans-11 CLA, of milk obtained from conventional and organic dairy sheep and goats farms under the farming conditions practiced in Greece. Four dairy sheep and four dairy goat farms, representing common conventional production systems and another four dairy sheep and four dairy goat farms, organically certified, representing organic production and feeding systems were selected from all over Greece. One hundred and sixty two individual milk samples were collected from those farms in January-February 2009, about three months after parturition. The milk samples were analyzed for their main chemical constituents and their FA profile. The results showed that the production system affected milk chemical composition: in particular fat content was lower in the organic sheep and goats milk compared with the corresponding conventional. Milk from organic sheep had higher content in MUFA, PUFA, alpha-LNA, cis-9, trans-11 CLA, and omega-3 FA, whereas in milk from organic goats alpha-LNA and omega-3 FA content was higher than that in conventional one. These differences are, mainly, attributed to different feeding practices used by the two production systems. The results of this study show that the organic milk produced under the farming conditions practiced in Greece has higher nutritional value, due to its FA profile, compared with the respective conventional milk.

Feed supplementation of Lactobacillus plantarum PCA 236 modulates gut microbiota and milk fatty acid composition in dairy goats--a preliminary study.

This study aimed to evaluate the potential of a promising Lactobacillus plantarum isolate (PCA 236) from cheese as a probiotic feed supplement in lactating goats. The ability of L. plantarum to survive transit through the goat gastrointestinal tract and to modulate selected constituents of the gut microbiota composition, monitored at faecal level was assessed. In addition, L. plantarum effects on plasma immunoglobulins and antioxidant capacity of the animals as well as on the milk fatty acid composition were determined. For the purpose of the experiment a field study was designed, involving 24 dairy goats of the Damascus breed, kept in a sheep and goat dairy farm. The goats were divided in terms of body weight in two treatments of 12 goats each, namely: control (CON) without addition of L. plantarum and probiotic (PRO) treatment with in feed administration of L. plantarum so that the goats would intake 12 log CFU/day. The experiment lasted 5 weeks and at weekly time intervals individual faecal, blood and milk samples were collected and analysed. All faecal samples were examined for the presence of L. plantarum PCA 236. In addition, the culturable population levels of mesophilic aerobes, coliforms lactic acid bacteria (LAB), Streptococcus, Enterococcus, mesophilic anaerobes, Clostridium and Bacteroides in faeces were also determined by enumeration on specific culture media. In parallel, plasma IgA, IgM and IgG and antioxidant capacity of plasma and milk were determined. No adverse effects were observed in the animals receiving the lactobacillus during the experiment. Lactobacillus plantarum PCA 236 was recovered in the faeces of all animals in the PRO treatment. In addition, PRO treatment resulted in a significant (P

Sheep and goats differences in CLA and fatty acids milk fat content in relation with mRNA stearoyl-CoA desaturase and lipogenic genes expression in their mammary gland.

An experiment was conducted with 12 lactating dairy ewes and 12 goats with the objective to determine whether, under the same dietary treatments, the differences in their fatty acid (FA) profile with emphasis on cis-9 trans-11 CLA milk fat content, are reflected in the transcript levels of genes involved in FA and cis-9, trans-11 CLA biosynthesis. The animals were fed with two diets (A, B) in different days of milk (DIM) due to the different milk yield, body weight etc, in order to have the same food intake and to avoid dietary effects. Diet A was fed to the animals on a group basis as it is traditionally used in practice, while diet B was chosen to avoid individual feed intake variation which is usually observed in group feeding. The results showed that there are significantly lower mRNA levels of acetyl-CoA carboxylase (ACC) in sheep mammary gland compared with those of goats, independently from the diet fed. The same trend was observed with the mRNA level of FA synthase (FAS), but the results were significant only for diet A. The mRNA level of lipoprotein lipase (LPL) in the mammary gland did not differ between sheep and goats fed with diet A. In addition, the concentration of cis-9 trans-11 CLA content was significantly higher in sheep milk fat compared with those of goats. This is in accordance with the significant higher levels on mRNA of stearoyl-CoA desaturase (SCD) which were observed in their mammary adipocytes of sheep compared with those of goats, independently of the fed diet (A or B). In conclusion, these findings demonstrate that the differences between sheep and goats, concerning cis-9, trans-11 CLA and FA milk fat content, under the same dietary treatments could be explained in part by the differences in mRNA of SCD and lipogenic genes in their mammary gland.

Effect of selenium supplementation on thyroid hormone levels and selenoenzyme activities in growing lambs.

The aim of the present study was to evaluate the effects of selenium supplementation on thyroid hormone metabolism and selenoenzyme activities in lambs. Twelve 20-d-old male lambs were assigned to one of two diets: A (0.11 ppm Se) and B (supplemented with 0.2 ppm selenium as sodium selenite). Blood samples were collected weekly for the determination of T3, T4, and selenium levels. The response to thyrotropin-releasing hormone (TRH) challenge was estimated at the 11th and 20th wk. Animals were slaughtered at wk 20 and tissues were collected for enzyme determination. Plasma selenium concentration was significantly higher in supplemented lambs (p<0.001). Plasma T3 and T4 levels remained similar in both groups. Type I deiodinase activity (ID-I) was decreased in the liver (p<0.05) and increased in the pituitary (p<0.01) of supplemented animals. No ID-I activity was detected in the thyroid. Pituitary type II deiodinase activity (ID-II) remained unchanged. The response to TRH challenge did not differ between the two groups for both challenges, but in group B, the second TRH challenge (20th wk) resulted in a significantly higher T3 response compared to the first one (11th wk) (p<0.05). In conclusion, the lack of effects of Se supplementation on thyroid hormone metabolism demonstrates that enzyme activity is homeostatically controlled and selenium is incorporated in that order to ensure the maintenance of thyroid hormone homeostasis.

Selenoenzyme activities in selenium- and iodine-deficient sheep.

This study was conducted to evaluate the effects of single and combined deficiencies of selenium and iodine on selenoenzyme activities in sheep. Twenty-four male lambs were assigned to one of four semisynthetic diets: combined deficient A (Se-I), Se-deficient B (Se-I+), I-deficient C (Se+I-), and basal diet D (Se+I+). Thyroid hormones (T3, T4), thyroid stimulating hormone (TSH), and inorganic iodine (PII) were determined in plasma. Selenium and glutathione peroxidase activity (GSH-Px) were determined in erythrocytes, and tissue samples, including the thyroid, liver, kidney, and brain, were taken for selenoenzyme analysis. Plasma T3, T4, and TSH concentrations were similar in all groups. Type I deiodinase (ID-I) activity in liver and kidney remained unchanged in Se or I deficiency. In contrast, hepatic ID-I activity was increased by 70% in combined Se-I deficiency. Thyroidal cystolic GSHPx (c-GSH-Px) and phospholipid GSH-Px (ph-GSH-Px) activities remained constant in both Se-deficient groups, whereas thyroidal c-GSH-Px activity increased (57%) in I deficiency. Type II deiodinase (ID-II) activity was not detectable in the cerebrum and cerebellum, whereas cerebellum Type III deiodinase (ID-III) activity was decreased in I deficiency and combined Se-I deficiencies. The results of the present study support a sensitive interaction between Se and I deficiencies in sheep thyroid and brain. Furthermore, the lack of thyroidal ID-I activity, the preservation of the thyroidal antioxidant enzymes, and the increases in hepatic ID-I indicate that a compensatory mechanism(s) works toward retaining plasma T3 levels, mostly by de novo synthesis of T3 and peripheral deiodination of T4 in Se- and I-deficient sheep.