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.

Microalgae as a Sustainable Source of Antioxidants in Animal Nutrition, Health and Livestock Development.

Microalgae are a renewable and sustainable source of bioactive compounds, such as essential amino acids, polyunsaturated fatty acids, and antioxidant compounds, that have been documented to have beneficial effects on nutrition and health. Among these natural products, the demand for natural antioxidants, as an alternative to synthetic antioxidants, has increased. The antioxidant activity of microalgae significantly varies between species and depends on growth conditions. In the last decade, microalgae have been explored in livestock animals as feed additives with the aim of improving both animals' health and performance as well as product quality and the environmental impact of livestock. These findings are highly dependent on the composition of microalgae strain and their amount in the diet. The use of carbohydrate-active enzymes can increase nutrient bioavailability as a consequence of recalcitrant microalgae cell wall degradation, making it a promising strategy for monogastric nutrition for improving livestock productivity. The use of microalgae as an alternative to conventional feedstuffs is becoming increasingly important due to food-feed competition, land degradation, water deprivation, and climate change. However, the cost-effective production and use of microalgae is a major challenge in the near future, and their cultivation technology should be improved by reducing production costs, thus increasing profitability.

Compositional Differences of Greek Cheeses of Limited Production.

Greece has a long tradition in cheesemaking, with 22 cheeses registered as protected designation of origin (PDO), 1 as protected geographical indication (PGI), and 1 applied for PGI. Several other cheeses are produced locally without any registration, which significantly contribute to the local economy. The present study investigated the composition (moisture, fat, salt, ash, and protein content), color parameters, and oxidative stability of cheeses that do not have a PDO/PGI certification, purchased from a Greek market. Milk and cheese types were correctly assigned for 62.8 and 82.1 % of samples, respectively, through discriminant analysis. The most important factors for milk type discrimination were L, a and b color attributes, salt, ash, fat-in-dry-matter, moisture-in-non-fat-substance, salt-in-moisture, and malondialdehyde contents, whereas a and b, and moisture, ash, fat, moisture-in-non-fat substance contents, and pH were the most influential characteristics for sample discrimination according to cheese type. A plausible explanation may be the differences in milk chemical composition between three animal species, namely cows, sheep, and goats and for the manufacture procedure and ripening. This is the very first report on the proximate analysis of these, largely ignored, chesses aiming to simulate interest for further study and production valorization.

Inclusion of Camelina sativa Seeds in Ewes' Diet Modifies Rumen Microbiota.

Supplementing ruminant diets with unconventional feedstuffs (Camelina sativa seeds; CS) rich in bioactive molecules such as polyunsaturated fatty acids, may prove a potential eco-efficient strategy to manipulate rumen microbiome towards efficiency. Forty-eight ewes were divided into four homogenous groups (n = 12) according to their fat-corrected milk yield (6%), body weight, and age, and were fed individually with concentrate, alfalfa hay, and wheat straw. The concentrate of the control group (CON) had no CS inclusion, whereas the treated groups were supplemented with CS at 60 (CS6), 110 (CS11), and 160 (CS16) g·kg-1 of concentrate, respectively. Rumen digesta was collected using an esophageal tube and then liquid and solid particles were separated using cheesecloth layers. An initial bacteriome screening using next-generation sequencing of 16S was followed by specific microbes targeting with a RT-qPCR platform, which unveiled the basic changes of the rumen microbiota under CS supplementation levels. The relative abundances of Archaea and methanogens were significantly reduced in the solid particles of CS11 and CS16. Furthermore, the relative abundance of Protozoa was significantly increased in both rumen fluid and solid particles of the CS6, whereas that of Fungi was significantly reduced in the rumen particle of the CS16. In rumen fluid, the relative abundance of Fibrobacter succinogens and Ruminobacter amylophilus were significantly increased in the CS6 and CS11, respectively. In the solid particles of the CS11, the relative abundance of Ruminococcus flavefaciens was significantly reduced, whereas those of Butyrivibrio proteoclasticus and Ruminobacter amylophilus were significantly increased. Additionally, the relative abundance of Selenomonas ruminantium was significantly increased in both CS11 and CS16. Consequently, the highest CS content in the concentrate reduced the relative abundance of methanogens without inducing radical changes in rumen microorganisms that could impair ruminal fermentation and ewes' performance.

Effect of Spirulina Dietary Supplementation in Modifying the Rumen Microbiota of Ewes.

Supplementing ruminant diets with microalgae, may prove an effective nutritional strategy to manipulate rumen microbiota. Forty-eight ewes were divided into four homogenous groups (n = 12) according to their fat-corrected milk yield (6%), body weight, age, and days in milk, and were fed individually with concentrate, alfalfa hay, and wheat straw. The concentrate of the control group (CON) had no Spirulina supplementation, while in the treated groups 5 (SP5), 10 (SP10), and 15 g (SP15) of Spirulina were supplemented as an additive in the concentrate. An initial screening using metagenomic next-generation sequencing technology was followed by RT-qPCR analysis for the targeting of specific microbes, which unveiled the main alterations of the rumen microbiota under the Spirulina supplementation levels. The relative abundance of Eubacterium ruminantium and Fibrobacter succinogenes in rumen fluid, as well as Ruminococcus albus in rumen solid fraction, were significantly increased in the SP15 group. Furthermore, the relative abundance of Prevotella brevis was significantly increased in the rumen fluid of the SP5 and SP10 groups. In contrast, the relative abundance of Ruminobacter amylophilus was significantly decreased in the rumen fluid of the SP10 compared to the CON group, while in the solid fraction it was significantly decreased in the SP groups. Moreover, the relative abundance of Selenomonas ruminantium was significantly decreased in the SP5 and SP15 groups, while the relative abundance of Streptococcus bovis was significantly decreased in the SP groups. Consequently, supplementing 15 g Spirulina/ewe/day increased the relative abundance of key cellulolytic species in the rumen, while amylolytic species were reduced only in the solid fraction.

Schizochytrium spp. Dietary Supplementation Modulates Immune-Oxidative Transcriptional Signatures in Monocytes and Neutrophils of Dairy Goats.

The high propensity of dietary polyunsaturated fatty acids (PUFA) to oxidation can induce a cascade of cellular immune-oxidative imbalances. On the other hand, PUFA, namely docosapentaenoic acid (ω6-DPA) and docosahexaenoic acid (DHA) can exert immunomodulatory effects by suppressing a pro-inflammatory response. Thus, the objective of this study was to investigate the effect of dietary supplementation with Schizochytrium spp. levels, rich in both ω6-DPA and DHA on the transcriptional profiling of genes involved in oxidative homeostasis and innate immunity of dairy goats' monocytes and neutrophils. Twenty-four dairy goats were divided into four homogeneous sub-groups; the diet of the control group (CON) had no Schizochytrium spp. while those of the treated groups were supplemented daily with 20 (ALG20), 40 (ALG40), and 60 (ALG60) g/goat/day. The mRNA levels of MGST1 in neutrophils were downregulated (p = 0.037), while in monocytes, SOD2 and SOD3 were downregulated (p = 0.010 and p = 0.044, respectively) in ALG60 compared to the CON group. GPX2 mRNA levels were downregulated (p = 0.036) in ALG20 and ALG60 compared to the CON group in neutrophils. NOX1 was upregulated (p = 0.043) in the neutrophiles of ALG60-goats. NOX2 was upregulated (p = 0.042) in the monocytes of ALG40-fed goats, while higher (p = 0.045) levels were also found in the ALG60 group in neutrophils. The mRNA levels of COX2 were downregulated (p = 0.035) in monocytes of the ALG40 and ALG60 groups. The mRNA levels of PTGER2 were also downregulated (p = 0.004) in monocytes of Schizochytrium-fed goats, while in neutrophils, significant downregulation (p = 0.024) was only found for ALG60 compared to the CON group. ALOX5AP mRNA levels were significantly decreased (p = 0.033) in ALG60 compared to the CON group in monocytes. LTA4H mRNA levels were increased (p = 0.015) in ALG60 compared to ALG20 and ALG40 groups in monocytes, while in neutrophils, a significant downregulation (p = 0.028) was observed in ALG20 compared to the CON group. The inclusion of more than 20 g Schizochytrium spp./day in goats' diet induced imbalances in mechanisms that regulate the antioxidant system, while downregulated the expression of pro-inflammatory pathways in monocytes and neutrophils.

The Effect of Dietary Inclusion of Microalgae Schizochytrium spp. on Ewes' Milk Quality and Oxidative Status.

An unprecedented challenge for nutritionists arises during the 21st century in order to produce highly nutritious and functional food which promotes human health. Polyunsaturated fatty acids (PUFA) that are highly contained in microalgae have broadly been confirmed for preventing cardiovascular diseases and regulating immune-oxidative status. However, their optimum dietary inclusion level needs to be defined since PUFA are prone to oxidation. For this purpose, 24 cross-bred dairy ewes, were separated into four groups (n = 6) and were fed with different levels of microalgae Schizochytrium spp. [0 (CON, no microalgae), 20 (SC20), 30 (SC30) and 40 (SC40) g/ewe/day] for 60 days. The results showed that although the production parameters were not impaired, milk fat content was decreased in medium and high-level supplemented groups while protein content was suppressed only for the medium one. Concerning the fatty acids (FA) profile, the proportions of C14:0, trans C18:1, trans-11 C18:1, cis-9, trans-11 C18:2, trans-10, cis-12 C18:2, C20:5 (EPA), C22:5n-6 (DPA), C22:6n-3 (DHA), the total ω3 FA and PUFA were significantly increased, while those of C18:0, cis-9 C18:1 and C18:2n-6c were decreased in the milk of treated ewes. Additionally, in the S40 group an oxidative response was induced, observed by the increased malondialdehyde (MDA) levels in milk and blood plasma. In conclusion, the dietary inclusion of 20 g Schizochytrium spp./ewe/day, improves milks' fatty acid profile and seems to be a promising way for producing ω3 fatty acid-enriched dairy products.

Consumer attitudes toward dairy products from sheep and goats: A cross-continental perspective.

This study aimed to assess consumer knowledge, attitudes, and perceptions toward dairy products from sheep and goats. A web-based survey was conducted in Latin America (Mexico and Chile), Europe (Italy, Spain, Greece, and Denmark), and Asia (Bangladesh). From March to June 2021, adult participants answered an online survey available in 5 languages. In total, 1,879 surveys were completed. Categorical and ordinal data were analyzed as frequencies and percentages. To determine the relationship between the variables for purchasing and consumption behaviors of respondents who declared that they consume dairy products, a multiple correspondence analysis was carried out. Most completed surveys were from Mexico and Italy (30% and 33.7%, respectively). Most respondents were between 18 and 29 yr old, female, highly educated, and employed. The majority of respondents (70.8%) declared that they consume dairy products from small ruminants. Consumers preferred products from both sheep and goats (49.4%); however, it was observed that in Mexico, Denmark, and Bangladesh, more than 50% preferred goat dairy products. The most-consumed products were mature and fresh cheeses. Mature cheese was the most-preferred product in Chile; in Mexico, Italy, Greece, and Denmark, it was fresh cheese. Unlike the rest of the countries, in Bangladesh, dairy product consumption from small ruminants was observed by more than 30% of respondents. In Mexico, a higher percentage of people do not consume sheep or goat dairy products because they are unfamiliar with them. In Mexico, Chile, and Bangladesh, limited market availability was also a variable responsible for nonconsumption. In European and Asian countries, sheep and goat dairy products are not consumed because consumers dislike them, in addition to a greater awareness of sustainability and climate change issues. The multiple correspondence analysis defined 5 dimensions. Dimension 1 was associated with the geographic location of the respondent (country and continent), the type of milk (sheep or goat), and the consideration of well-being and health as characteristics associated with the consumption of dairy products from small ruminants. Dimension 2 was associated with the respondent's country of origin and the frequency of consumption. Dimension 3 was associated with gender, education, and employment status. Dimension 4 was associated with the respondent's age, the association of the "healthy" concept of sheep and goat dairy products, and the consideration of the nutritional benefits of dairy as responsible for considering them healthy. Dimension 5 was associated with a "strong smell and taste" of sheep and goat dairy products. This study showed that consumer attitudes toward dairy products from sheep and goats vary between continents. In conclusion, results showed consumer interest in animal welfare and environmental impact issues related to small ruminant farming as well as a general attraction to local products. It seems that these factors contribute to consumers' perception of the quality of dairy products, so the industry and select farmers should carefully consider incorporating them into their supply chain.

Long-term administration of a commercial supplement enriched with bioactive compounds does not affect feed intake, health status, and growth performances in beef cattle.

Feed additives including natural bioactive compounds (BCs) in combination with vitamin E (VitE) and organic Se could mitigate animal stress associated with intensive livestock farming due to their anti-inflammatory and antioxidant properties. Yeast and yeast derivate are included in feed additives as probiotic products and digestion promoters. Scutellaria baicalensis is a source of bioactive compounds and has been tested in monogastrics, exhibiting many immunostimulating and hepato-protective activities. However, the literature lacks information regarding S. baicalensis effects on beef cattle performance and health status. The aim of the present study was to evaluate the impact on beef cattle's feed intake, health and oxidative status, and growth performances of the inclusion of a commercial supplement (CS) containing VitE, organic Se, yeast derivate, and S. baicalensis extract during the fattening and finishing period. A total of 143 Charolaise male cattle were allotted into 12 pens of 11-12 animals each and assigned to a control ( 463.9 ± 21.48 body weight - BW) or a treated ( 469.8 ± 17.91  BW) group. Each group included two replicates of three pens. The treated groups were supplemented with 20  g CS animal - 1 d - 1 . Feed intake was measured monthly on a pen base during two consecutive days. Total mixed ration and fecal samples were collected at three time points (monthly, from November to February) and pooled by replicate for the analyses to monitor digestibility. Blood samples were individually collected at the beginning and at the end of the trial for oxidative status and metabolic profile determination. Final BW and carcass weight were individually recorded to calculate average daily gain, feed conversion ratio, and carcass yield. Similar feed digestibility between groups were observed during the whole experiment. Feed intake, growth performances, final body weight, average daily gain, feed conversion rate, oxidative status, and metabolic profile were not affected by the dietary inclusion of the tested CS indicating no detrimental effect of the treatment. Different doses of this product should be tested in the future in order to provide a more complete report on the product efficacy.

Effect of a Carotenoid Extract from Citrus reticulata By-Products on the Immune-Oxidative Status of Broilers.

Although carotenoids generally possess antimicrobial and antioxidant properties, the in vivo synergistic action of carotenoid blends derived from plant-based by-products has not been thoroughly studied. Therefore, the carotenoid characterization and antimicrobial potential of Citrus reticulata extract as well as the impact of this carotenoid-rich extract (CCE) dietary supplementation on the performance, meat quality, and immune-oxidative status of broiler chickens were determined. One hundred and twenty one-day-old hatched chicks (Ross 308) were allocated to two dietary groups, with four replicate pens of 15 birds each. Birds were fed either a basal diet (CON) or the basal diet supplemented with 0.1% CCE (25 mg carotenoid extract included in 1 g of soluble starch) for 42 d. ß-Cryptoxanthin, ß-Carotene, Zeaxanthin, and Lutein were the prevailing carotenoid compounds in the Citrus reticulata extract. The CCE feed additive exerted inhibitory properties against both Gram-positive (Staphylococcus aureus) and negative (Klebsiella oxytoca, Escherichia coli, and Salmonella typhimurium) bacteria. Both the broiler performance and meat quality did not substantially differ, while the breast muscle malondialdehyde (MDA) concentration tended to decrease (p = 0.070) in the CCE-fed broilers. The inclusion of CCE decreased the alanine aminotransferase and MDA concentration, and the activity of glutathione peroxidase, while the activity of superoxide dismutase was increased in the blood. Catalase and NADPH oxidase 2 relative transcript levels were significantly downregulated in the livers of the CCE-fed broilers. Additionally, Interleukin 1ß and tumor necrosis factor (TNF) relative transcript levels were downregulated in the livers of the CCE- fed broilers, while TNF and interferon γ (IFNG) tended to decrease in the spleens and bursa of Fabricius, respectively. The present study provided new insights regarding the beneficial properties of carotenoids contained in Citrus reticulata in broilers' immune-oxidative status. These promising outcomes could be the basis for further research under field conditions.

Effects of PUFA-Rich Dietary Strategies on Ruminants' Mammary Gland Gene Network: A Nutrigenomics Review.

Although the inclusion of polyunsaturated fatty acids (PUFAs) in ruminants' diets appears to be a well-documented strategy to enrich milk with PUFAs, several gene networks that regulate milk synthesis and mammary gland homeostasis could be impaired. The objective of this literature review is to assess the effects of nutritional strategies focused on enriching milk with PUFAs on gene networks regulating mammary gland function and lipogenesis, as well as the impact of feed additives and bioactive compounds with prominent antioxidant potential on immune-oxidative transcriptional profiling, as a part of mammary gland homeostasis and health. The findings support the conclusion that PUFAs' inclusion in ruminants' diets more strongly downregulate the stearoyl-CoA desaturase (SCD) gene compared to other key genes involved in de novo fatty acid synthesis in the mammary gland. Additionally, it was revealed that seed oils rich in linoleic and linolenic acids have no such strong impact on networks that regulate lipogenic homeostasis compared to marine oils rich in eicosapentaenoic and docosahexaenoic acids. Furthermore, ample evidence supports that cows and sheep are more prone to the suppression of lipogenesis pathways compared to goats under the impact of dietary marine PUFAs. On the other hand, the inclusion of feed additives and bioactive compounds with prominent antioxidant potential in ruminants' diets can strengthen mammary gland immune-oxidative status. Considering that PUFA's high propensity to oxidation can induce a cascade of pro-oxidant incidences, the simultaneous supplementation of antioxidant compounds and especially polyphenols may alleviate any side effects caused by PUFA overload in the mammary gland. In conclusion, future studies should deeply investigate the effects of PUFAs on mammary gland gene networks in an effort to holistically understand their impact on both milk fat depression syndrome and homeostatic disturbance.

Immune-Related Gene Expression Profiling of Broiler Chickens Fed Diets Supplemented with Vinification Byproducts: A Valorization Approach II.

The valorization of vinification byproducts portrays a promising bioprocess for the enrichment of animals' diet with bioactive compounds, such as polyphenols, which could regulate the immune response. Therefore, the impact of dietary grounded grape pomace (GGP), wine lees extract (WYC), and grape stem extract (PE) on the relative transcript level of immune related genes of broiler chickens were examined. Two hundred forty, one-day-old as hatched (male/female) chicks (Ross 308) were allocated to four dietary groups, with four replicate pens each with 15 birds. Birds were fed either a basal diet (CON) or the basal diet supplemented with 2.5% GGP, or 0.2% WYC, or 0.1% PE for 42 d. The relative expression of immune-related genes was investigated using a real-time PCR platform. The mRNA levels of Toll-like Receptor 4 (TLR4) were downregulated (p = 0.039) in the liver of broilers fed the GGP-containing diet compared to the CON, while in the spleen of PE-fed broilers, TLR4 was significantly upregulated (p = 0.043). The mRNA levels of interleukin 8 (IL8) tended to upregulate (p = 0.099) in the bursa of Fabricius and were significantly increased (p = 0.036) in the spleen of broilers fed the PE diet. Vinification byproducts depict a promising sustainable source of polyphenols for the poultry feed industry, but more research is needed under field conditions.

Plant Feed Additives as Natural Alternatives to the Use of Synthetic Antioxidant Vitamins on Livestock Mammals' Performances, Health, and Oxidative Status: A Review of the Literature in the Last 20 Years.

In the last two decades, the interest in natural plant feed additives (PFA) as alternatives to synthetic vitamins in livestock nutrition has increased. After a systematic review, a total of 19 peer-reviewed papers published between 2000 and 2020 were retained to evaluate the antioxidant effects of PFA compared to synthetic antioxidant vitamins (mainly vitamin E; VitE) in livestock nutrition. These studies demonstrated that PFAs could be as efficient as VitE in counteracting oxidative stress in pigs, rabbits, and ruminants. However, PFAs only positively affected animals' growth performance and feed efficiency in some monogastric studies. The PFA can affect antioxidant enzyme activity in a dose- and method of administration-dependent manner. The antioxidant capacity of both PFA and VitE were depressed in cows fed with diets rich in polyunsaturated fatty acids. Variability among studies could be related to species differences. Despite the interest of the feed industry sector in PFA, there are still very few studies evaluating their antioxidant effect in species other than poultry.

The Effect of Forage-to-Concentrate Ratio on Schizochytrium spp.-Supplemented Goats: Modifying Rumen Microbiota.

The inclusion of feed additives and the implementation of various nutritional strategies are studied to modify the rumen microbiome and consequently its function. Nevertheless, rumen enzymatic activity and its intermediate products are not always matched with the microbiome structure. To further elucidate such differences a two-phase trial using twenty-two dairy goats was carried out. During the first phase, both groups (20HF n = 11; high forage and 20HG n = 11; high grain) were supplemented with 20 g Schizochytrium spp./goat/day. The 20HF group consumed a diet with a forage:concentrate (F:C) ratio of 60:40 and the 20HG-diet consisted of a F:C = 40:60. In the second phase, the supplementation level of Schizochytrium spp. was increased to 40 g/day/goat while the F:C ratio between the two groups were remained identical (40HF n = 11; high forage and 40HG n = 11; high grain). By utilizing a next-generation sequencing technology, we monitored that the high microalgae inclusion level and foremost in combination with a high grains diet increased the unmapped bacteria within the rumen. Bacteroidetes and Prevotella brevis were increased in the 40HG -fed goats as observed by using a qPCR platform. Additionally, methanogens and Methanomassiliicoccales were increased in high microalgae-fed goats, while Methanobrevibacter and Methanobacteriales were decreased. Fibrolytic bacteria were decreased in high microalgae-fed goats, while cellulolytic activity was increased. Ammonia was decreased in high grains-fed goats, while docosapentaenoic and docosahexaenoic acids showed a lower degradation rate in the rumen of high forage-fed goats. The alteration of the F:C ratio in goats supplemented with Schizochytrium spp. levels modified both ruminal microbiota and enzymatic activity. However, there was no significant consistency in the relations between them.

Assessing the Optimum Level of Supplementation with Camelina Seeds in Ewes' Diets to Improve Milk Quality.

Camelina sativa seeds are rich in bioactive compounds such as polyunsaturated fatty acids (PUFA) and antioxidants, thus, their supplementation in ewes' diets, may be an effective way to develop high nutritional dairy products. Therefore, the present study investigates the effect of the dietary inclusion of Camelina sativa seeds in ewes' oxidative status and milk quality. Forty-eight dairy Chios ewes were divided into four homogenous groups and were fed individually. The concentrate of the control group (CON) had no inclusion of Camelina seeds, while the treatment groups (CSS6, CSS11, CSS16) were supplemented with 6%, 11%, and 16%, respectively. Including Camelina seeds in 6% and 11%, had no impact on milk performance, while in the CSS16, milk fat was significantly decreased compared to the CON. Supplementing Camelina seeds improved milk quality from a human health perspective by modifying the content of saturated fatty acid, the proportions of α-linolenic (C18:3 n-3), and C18:2 cis-9, trans-11 (CLA), and the ω6/ω3 ratio. Furthermore, the activity of catalase (CAT) was significantly increased in the CSS11 and CSS16, and superoxide dismutase (SOD) activity also significantly upsurged in the CSS16. Still, the levels of malondialdehyde (MDA) were significantly increased in the CSS11 compared to the CON and CSS6, and in the CSS16 compared to the CSS6. In CSS16, protein carbonyls were significantly increased. Finally, in the CSS-fed ewes, milk oxidative stability was fortified, as suggested by the modifications in the activities of SOD, CAT, and glutathione peroxidase (GSH-Px), in the antioxidant capacity, and the oxidative stress biomarkers. Consequently, the incorporation of 6% Camelina seeds in the concentrates of ewes improves milk's fatty acid profile and oxidative status. However, more research is required regarding the possible negative effects of the constant consumption of Camelina seeds by ewes.

Antioxidant Status of Broiler Chickens Fed Diets Supplemented with Vinification By-Products: A Valorization Approach.

Vinification by-products display great potential for utilization as feed additives rich in antioxidant compounds. Thus, the effect of dietary ground grape pomace (GGP), wine lees extract rich in yeast cell walls (WYC), and grape stem extracts (PE) on the relative expression of several genes involved in liver oxidative mechanisms and the oxidative status of the blood and breast muscle of broiler chickens was investigated. In total, 240 one-day-old as hatched chicks (Ross 308) were assigned to four treatments, with four replicate pens and 15 birds in each pen. Birds were fed either a basal diet (CON) or a basal diet supplemented with 25 g/kg GGP, or 2 g/kg WYC, or 1 g starch including 100 mg pure stem extract/kg (PE) for 42 days. The polyphenolic content of vinification by-products was determined using an LC-MS/MS library indicating as prevailing compounds procyanidin B1 and B2, gallic acid, caftaric acid, (+)-catechin, quercetin, and trans-resveratrol. Body weight and feed consumption were not significantly affected. The relative transcript level of GPX1 and SOD1 tended to increase in the liver of WYC-fed broilers, while NOX2 tended to decrease in the PE group. SOD activity in blood plasma was significantly increased in WYC and PE compared to the CON group. The total antioxidant capacity measured with FRAP assay showed significantly higher values in the breast muscle of PE-fed broilers, while the malondialdehyde concentration was significantly decreased in both WYC- and PE-fed broilers compared to the CON group. The exploitation of vinification by-products as feed additives appears to be a promising strategy to improve waste valorization and supply animals with bioactive molecules capable of improving animals' oxidative status and products' oxidative stability.

Changes in the Rumen Bacteriome Structure and Enzymatic Activities of Goats in Response to Dietary Supplementation with Schizochytrium spp.

With the aim to produce functional dairy products enriched with polyunsaturated fatty acids (PUFA) by using feed supplements, radical changes could occur in the rumen microbiome. This work investigated the alterations of the rumen bacteriome of goats fed with PUFA-rich marine microalgae Schizochytrium spp. For the trial, twenty-four goats were divided into four homogenous clusters (six goats/treatment) according to their fat-corrected (4%) milk yield, body weight, and age; they were individually fed with alfalfa hay and a concentrate (F/C = 50/50). The concentrate of the control group (CON) contained no microalgae, while those of the treated groups were supplemented daily with 20 (ALG20), 40 (ALG40), and 60 g (ALG60) of Schizochytrium spp./goat. Rumen fluid samples were collected using a stomach tube during the 20th and 40th days of the experiment. The microbiome analysis using a 16S rRNA sequencing platform revealed that Firmicutes were decreased in microalgae-fed goats, while Bacteroidetes showed a tendency to increase in the ALG40 group due to the enhancement of Prevotellaceae. Cellulolytic bacteria, namely Treponema bryantii, Ruminococcus gauvreauii, R. albus, and R. flavefaciens, were decreased in the ALG40 group, resulting in an overall decrease of cellulase activity. In contrast, the amylolytic potential was significantly enhanced due to an upsurge in Ruminobacter amylophilus, Succinivibrio dextrinosolvens, and Fretibacterium fastidiosum populations. In conclusion, supplementing goats' diets with 20 g Schizochytrium spp. could be considered a sustainable and efficient nutritional strategy to modulate rumen microbiome towards the development of dairy products enriched with bioactive compounds, while higher levels induced substantial shifts in determinant microbes' populations.

Schizochytrium sp. Dietary supplementation modify Toll-like receptor 4 (TLR4) transcriptional regulation in monocytes and neutrophils of dairy goats.

Animals protect themselves against pathogens or abiotic factors by innate or adaptive mechanisms. Long-chain polyunsaturated fatty acids (ω3) of microalgae modify both human and mice' immune systems resulting in a beneficial balance between pro-inflammatory and anti-inflammatory pathways. However, scarce information exists on their impact on lactating animals' immunity. The objective of this study was to investigate the impact of dietary inclusion of Schizochytrium sp. (rich in docosapentaenoic and docosahexaenoic acid), on the expression of several genes involved in the innate immunity of goats. Twenty-four dairy goats were divided into four homogeneous sub-groups (n = 6). All goats were fed individually with alfalfa hay and concentrate. The concentrate of the control group (CON) had no microalgae while those of the treated groups were supplemented daily with 20 (ALG20), 40 (ALG40), and 60 (ALG60) g Schizochytrium sp. Monocytes and neutrophils were isolated from goats' blood in the 20th, 40th, and 60th days from the beginning of the experimental period. The relative transcript levels of TLR4, MYD88, MAPK, IRF3, IFNG, and pro-inflammatory cytokines (IL1B, IL2, IL8, TNF), and chemokines (CCL5 and CXCL16) were decreased in monocytes of microalgae treated goats compared to the CON. In contrast, MAPK and IL1B relative transcript levels were increased in neutrophils of ALG40 and ALG60 groups. In conclusion, the supplementation of goats' diet with 20 g Schizochytrium sp. resulted in a downregulation of the pro-inflammatory transcriptions, and following further research could be considered as a sustainable alternative strategy to improve immune function.

Effects of Inclusion of Schizochytrium spp. and Forage-to-Concentrate Ratios on Goats' Milk Quality and Oxidative Status.

Although the dietary inclusion level of polyunsaturated fatty acids (PUFA) and the forage: concentrate (F:C) ratio affect milk quality, their interaction has not been broadly studied. To address such gaps and limitations a two-phase trial using twenty-two dairy goats was carried out. During the first phase, both groups (20 HF n = 11; high forage and 20 HG n = 11; high grain) were supplemented with 20 g Schizochytrium spp./goat/day. The 20 HF group consumed a diet with F:C ratio 60:40 and the 20 HG-diet consisted of F:C = 40:60. In the second phase, the supplementation level of Schizochytrium spp. was increased to 40 g/day/goat while the F:C ratio between the two groups were remained identical (40 HF n = 11; high forage and 40 HG n = 11; high grain). Neither the Schizochytrium spp. supplementation levels (20 vs. 40) nor the F:C ratio (60:40 vs. 40:60) affected milk performance. The high microalgae level (40 g) in combination with high grain diet (40 HG) modified the proportions of docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), and conjugated linoleic acid (CLA) and the ω3/ω6 ratio in milk, to a beneficial manner according to human health recommendation guidelines. However, the highest inclusion level of Schizochytrium spp. (40 g) and foremost in combination with the high grain diets (40 HG) induced an oxidative response as observed by the increased protein carbonyls (CP) and malondialdehyde (MDA) levels in milk and blood plasma indicating severe limitations for a long-term, on-farm application. In conclusion, the supplementation with 20 g Schizochytrium spp. and high forage diet (60:40) appears to be an ideal formula to enrich dairy products with essential biomolecules for human health without adversely affect milk oxidative stability.

Effects of Supplementing Rumen-Protected Methionine and Lysine on Milk Performance and Oxidative Status of Dairy Ewes.

There is limited information on the impact of dietary supplementation with separate rumen-protected (RP) amino acids (AA), or with their combination, on ewes' oxidative status. Sixty ewes were divided into five groups; C: basal diet (control); M: basal diet + 6 g/ewe RP methionine; L: basal diet + 5 g/ewe RP lysine; LML: basal diet + 6 g methionine and 5 g lysine/ewe; and HML: basal diet + 12 g methionine + 5 g lysine/ewe. Milk's fat content increased in RP-AA fed ewes, while that of protein in M and L only. In blood plasma, the malondialdehyde (MDA) content was reduced in the M, LML, and HML compared to C-fed ewes. An increase in glutathione transferase activity in the blood plasma of the M and LML compared to the C and HML-fed ewes were found. In milk, lower values of the ferric reducing ability of plasma (FRAP) in the LML and HML-fed ewes and of 2,2'-Azino-bis 3-ethylbenzthiazoline-6-sulfonic acid (ABTS) in LML only, were found. Lysine increased milk's FRAP values and MDA content. Both L and HML diets increased milk's protein carbonyls content. Methionine improves the organism's oxidative status, without adversely affecting milk's oxidative stability. Lysine dietary inclusion affects negatively the oxidative stability of milk.