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This study aimed to evaluate the effect of hazelnut skin by-product supplementation on lamb meat quality characteristics and plasma and muscle proteomes. Twenty-two Valle del Belice male lambs were divided into two experimental groups: control (C), fed a maize-barley diet and hazelnut (H), fed hazelnut skin by-product as maize partial replacer in the concentrate diet. The meat of lambs fed hazelnut skin showed greater values of lightness, redness, yellowness, and chroma color parameters together with the highest myofibril fragmentation index. Two-dimensional electrophoresis and mass spectrometry applied on plasma proteome identified 20 protein spots corresponding to 18 unique gene names to be differently expressed due to hazelnut skin by-product substitution. For the early post-mortem muscle, 23 protein spots (42 unique gene names) were significantly up-regulated due to hazelnut skin by-product supplementation. Four proteins these being APOA1, PHB, ACTG1 and ALB, were found to be common to the two proteomes suggesting that these proteins could be candidate biomarkers to monitor in vita and post-mortem lamb meat quality traits. This study evidenced the main mechanisms involved in the supplementation of hazelnut skin by-product in lambs' diet and confirmed the possibility of using plasma proteome as a non-invasive way to predict lamb meat quality. SIGNIFICANCE: Maximizing the use of agro-industrial by-products as replacers of traditional feedstuff for improving animal products is one of the important challenges to preserving natural resources and guaranteeing environmental sustainability. Hazelnut (Corylus avellana L.) skin, obtained as a results of hazelnut roasting, represents a valuable by-products due to its high content in unsaturated fatty acids, tannins, and vitamins. Thus, including hazelnut skin by-product in small ruminant nutrition could reduce the costs of animal feedings for farmers as well as improve meat nutritional and sensorial characteristics. Additionally, monitoring the meat quality characteristics with fast, accurate, and non-invasive tools to find, before slaughter, animals with desired quality characteristics is of growing interest in the last years. In this regard, the objectives of this study were to assess i) the effect of hazelnut skin supplementation on lamb meat quality characteristics and plasma and muscle proteomes, and ii) whether analyzing plasma proteome by using a gel-based proteomic approach could effectively offer a more readily available option for determining lamb meat quality. Taken together, the proteomic approach applied to plasma and muscle proteomes, allowed us to reveal the pathways and the potential candidate plasma biomarkers to predict lamb meat production in the pre-slaughter phase.

Physical activity (PA) and sport play an essential role in promoting body development and maintaining optimal health status both in the short and long term. Despite the benefits, a long-lasting heavy training can promote several detrimental physiological changes, including transitory immune system malfunction, increased inflammation, and oxidative stress, which manifest as exercise-induced muscle damages (EIMDs). Meat and derived products represent a very good source of bioactive molecules such as proteins, lipids, amino acids, vitamins, and minerals. Bioactive molecules represent dietary compounds that can interact with one or more components of live tissue, resulting in a wide range of possible health consequences such as immune-modulating, antihypertensive, antimicrobial, and antioxidative activities. The health benefits of meat have been well established and have been extensively reviewed elsewhere, although a growing number of studies found a significant positive effect of meat molecules on exercise performance and recovery of muscle function. Based on the limited research, meat could be an effective post-exercise food that results in favorable muscle protein synthesis and metabolic performance.

Nonbovine milk products mainly from goat, sheep, camel, and donkey have recently attracted attention for their nutritional, health-promoting, and processing properties. The present chapter addresses an overview of the recent nonbovine milk market scenario, facing the critical aspects for growing nonbovine dairying, and illustrates the main differences among protein and fat composition of nonbovine milk. Moreover, the newest dairy products from sheep, goat, camel, and equid milk were presented in the light of the recent consumer’s request, among which probiotic and low-fat products, and emerging applications as ingredient in functional food, such as use of oligosaccharides from goat milk and novel probiotic strains isolated from camel milk.

In the present study, the evaluation of the carcasses and meat quality, in terms of chemical composition and fatty acid profile, of lambs from five autochthonous sheep breeds (Altamurana, Bagnolese, Gentile di Puglia, Laticauda, and Leccese) reared in continental Southern Italy, were studied. All the carcasses were evaluated according to the EU Mediterranean classification system for carcasses weighing less than 13 kg. Meat chemical composition and fatty acids profile were assessed on both loin and leg commercial cuts. Fatty acid composition of loin resulted in differences among breeds, displaying lower values of saturated fatty acid in Altamurana, Bagnolese, and Leccese breeds and the highest content of polyunsaturated fatty acid in the Altamurana breed. Principal component analysis grouped lamb according to fatty acid content and to conjugated linoleic acid (CLA), omega n-3 and n-6 fatty acids, thus, Altamurana, Bagnolese, and Leccese breeds are characterized by the highest values of CLA content. Our data demonstrated that lamb meat from autochthonous breed has good carcass quality and the content of CLA, n-3, and n-6 was valuable for human consumption, therefore, the valorisation of local meat quality can help to avoid the extinction of the autochthonous breed offering to the market and consumer’s high nutritive products.

Sheep milk contains a large number of antioxidant molecules, such as whey proteins, vitamins, fatty acids and others. In sheep milk the oxidation process is naturally balanced by the presence of constitutive antioxidant agents, however, the abundance of this antioxidant compounds depends also by the stressor conditions to which animal can be exposed. The stage of lactation together with particular environmental conditions, such as the high ambient temperatures, can cause an imbalance between oxidants and antioxidants, being responsible for the oxidative stress that can be measured both in plasma and in milk samples. Different methods to measure the antioxidant activity of milk have been proposed, according to each single antioxidant compounds and to the need for consumers’ health of decreasing the exposition to oxidized proteins capable for creation of a novel class of allergens. However, the golden standard for the measurement of antioxidant activity in milk has not still found. Therefore, the present review aims at describing the roles of various antioxidant molecules of milk, and provides an overview of the different methods available to measure the total antioxidant capacity of milk.

The objective of this paper was to assess the plasma antioxidant status ewes under high ambient temperature fed seaweed Ascophyllum nodosum (AG), flaxseed (FS) and their combination (FS + AG) measured by the Antioxidant/Oxidant Balance (AOB). The experiment lasted 30 day and involved thirty-two late-lactation Comisana ewes, divided into four balanced groups of eight each, and exposed to high ambient temperature. The ewes in each group were individually fed twice daily and received 1.8 kg per day of oat hay. Results showed that ewes fed the combination of flaxseed and A. nodosum displayed the highest AOB value at day 30 of high ambient temperature exposure; moreover, they displayed lower rectal temperature than ewes fed A. nodosum alone, and lower concentrations of heat shock protein 70. These data demonstrated that supplementation of diet with A. nodosum in combination with flaxseed can protect ewes from the excessive oxidative imbalance caused by high ambient temperature.

Neutrophils represent primary mobile phagocytes recruited to the site of infection, and their functions are essential to enhance animals' health performance. Neutrophils have an essential role in innate immunity and are able to kill the pathogens via the synthesis of neutrophil extracellular traps (NETs). The objective of the present work was the study of the in vitro NETosis of peripheral neutrophils isolated from dairy cows supplemented with olive pomace. Dairy cows (n = 16) balanced for parity (3.67 ± 1.5 for CON, 3.67 ± 1.9 for OP), milk yield (24.3 ± 4.5 kg d−1for CON and 24.9 ± 1.7 kg d−1 for OP), the number of days in milk (109 ± 83.5 for CON and 196 ± 51 for OP), and body weight (647 ± 44.3 kg for CON and 675 ± 70.7 kg for OP) were divided into two experimental groups fed with a control diet (CON) and supplemented with 6% of olive pomace (OP). Peripheral blood neutrophils were isolated and stimulated in vitro with phorbol-myristate-acetate (PMA) as a marker for activation and reactivity of the neutrophils. After isolation, both the viability and CD11b expression were analyzed by flow cytometry. Both NETosis by neutrophil elastase-DNA complex system and myeloperoxidase (MPO) activity were evaluated by ELISA. The specific antibodies against MPO and citrullination of Histone-H1 were used for investigating NETosis by immunofluorescence microscopy. The neutrophil elastase-DNA complexes produced during NETosis and MPO activity of neutrophil extracts were affected by OP supplementation. Furthermore, results from immunofluorescence analysis of NETosis depicted a similar result found by ELISA showing a higher expression of MPO and citrullination of Histone-H1 in OP than the CON neutrophils. In addition, all data showed that the OP diet resulted in a better response of neutrophils to PMA stimulation than the CON diet, which did not support the neutrophils' responses to PMA stimulation. Our results demonstrated that OP supplementation can enhance the neutrophil function in dairy cows leading to udder defense and inflammation response especially when an immunosuppression state can occur.

In the last ten years there is a growing concern about livestock health and welfare in relation to the spread of antimicrobial resistance caused by the misuse of antibiotics, highlighting the interest of new alternatives to antibiotics in veterinary medicine practices. A number of studies investigated the potential use of aromatic plants and their extracts, such as essential oils, as alternatives to the antibiotics use in ruminant nutrition. More recently, aromatic plant-derived bioactive compounds (ABIOC) have attracted the attention as alternatives to antibiotics for their potential antimicrobial activities and to growth promoters in livestock. In vivo and in vitro studies on dairy animals demonstrated that ABIOC could favorably alter ruminal fermentation stimulating the growth of beneficial microbes and limit the number of pathogenic bacteria; therefore, they could potentially improve the feed efficiency of ruminants and enhance the production and nutritive quality of milk. The effects of aromatic plants and their extracts in dairy animal will be discussed in the chapter.

The present experiment aimed at understanding the effects of cortisol levels on sheep peripheral blood mononuclear cell (PBMC) proliferation and cytokine production during hyperthermia. To mimic stress related to the exposition of high ambient temperatures, PBMC were cultured at 43°C for 12 h, and subsequently at 39°C for additional 12 h. Cells in normothermia were cultured at 39°C for 24 h. Phytohemagglutinin-stimulated PBMC were cultured with different cortisol levels: 0 ng/mL; 100 ng/mL, representing the physiological cortisol concentration simulating stress condition (Cort100); and 1,000 ng/mL, representing the hyperactivated hypothalamic-pituitary-adrenal axis (Cort1000). Phytohemagglutinin-stimulated PBMC with 0 ng/mL of cortisol concentration represented the positive control, whereas nonstimulated PBMC without cortisol represented the negative control (NC). The free cell supernatants were collected for the determination of IL-6, IL-1β, and IL-10 by ELISA. Bromodeoxyuridine assay was performed on cells to determine cell proliferation. Exposition to hyperthermia negatively affected cell proliferation, IL-6, IL-1β, and IL-10 concentrations in cell supernatants. The interaction of hyperthermia and cortisol level affected both cell proliferation and IL-10 production. Both PBMC proliferation and IL-10 production in positive control, Cort100, and Cort1000 decreased at 43°C as compared with 39°C NC. On average, the Cort100 treatment displayed higher concentrations of IL-6 than NC. The present experiment demonstrated that the action of cortisol concentration simulating stress condition on cell proliferation and cytokine production was a permissive/stimulatory action during normothermia, whereas it was a suppressive action during hyperthermia. These data confirmed that cortisol concentration simulating stress condition could have a role in the immune system of sheep via mediating cellular homeostasis in the condition of hyperthermia. The negative effects of hyperthermia on sheep immune responses were apparent when performing an immunological challenge.

We present a pilot study on the effects of milk protein fractions [αS1-casein (CN), αS2-CN, κ-CN, β-CN, and a mix of α-lactalbumin (α-LA) and β-lactoglobulin (β-LG)] from different animal species (bovine, ovine, and caprine) on pro- and anti-inflammatory cytokines and oxidative status in cultured peripheral blood mononuclear cells from children with generalized epilepsy. Peripheral blood mononuclear cells (PBMC) were obtained by density gradient from blood of 10 children with generalized epilepsy (5 males; mean age 33.6 ± 5.4 mo) and 10 controls (5 males; mean age 35.6 ± 6.8 mo). Children with epilepsy were grouped according to cytokine levels as follows: children with epilepsy having low levels of cytokines not different from those of control children (LL-EC); children with epilepsy having cytokine levels at least 5-fold higher (medium levels) than those of control children (ML-EC); and children with epilepsy having cytokine levels at least 10-fold higher (high levels) than those of control children (HL-EC). The production of tumor necrosis factor-α (TNF-α), IL-10, IL-6, and IL-1β was studied in cultured PBMC incubated with αS1-CN, αS2-CN, κ-CN, β-CN, and a mix of α-LA and β-LG from bovine, caprine, and ovine milks. The levels of reactive oxygen and nitrogen species (ROS/RNS) and catalase activity were assessed in cultured supernatant. In the HL-EC group, β-CN from small ruminant species (ovine and caprine) induced the highest levels of TNF-α, whereas PBMC incubated with αS2-CN from ovine milk and the mix of β-LG and α-LA from all tested milk species had the lowest levels of TNF-α. Within the HL-EC group, production of IL-1β was higher for bovine and ovine αS2-CN fractions and lower for caprine and ovine β-CN and κ-CN. In the HL-EC group, IL-6 was higher in cultured PBMC incubated with αS2-CN from bovine and ovine milk than from caprine milk. The cytokine IL-10 did not differ among milking species. The highest levels of ROS/RNS were found after incubation of PBMC with the β-CN fraction in bovine milk. Catalase activity was higher in PBMC cultured with β-CN isolated from bovine and caprine milk and with αS1-CN from ovine milk.

We report the fatty acid profile of raw milk and of the corresponding digested milk from different sources (human milk, formula milk and donkey, bovine, ovine and caprine milk) to gain information on the nutritional quality of different milk sources in infant nutrition.Short chain fatty acids (SC-FA) were higher in bovine and caprine milk, intermediate in ovine and donkey and lower in human and formula milk. Medium chain fatty acids (MC-FA) showed the highest values for bovine and caprine milk and the lowest for donkey and formula milk, whereas long chain fatty acids (LC-FA) were the highest in donkey and formula milk and intermediate in human milk.The percentage distribution of fatty acids liberated after in vitro digestion did not reflect the patterns found in the corresponding milk sources. In particular, MC free fatty acids (MC-FFA) showed the highest and the lowest values in donkey and in formula milk, LC-FFA showed the highest value in human milk. The total FFA was highest in human milk, lowest in formula milk and intermediate in donkey, bovine, ovine, and caprine milk.

In this study, the effects of melatonin treatment on growth, redox status and immunity in prenatally stressed newborn lambs were evaluated. Thirty-seven newborn lambs were allocated into two groups (melatonin-MEL and control-CON), based on whether their mothers were treated with melatonin implants or not, respectively. All pregnant ewes were exposed to heat stress. The body weight of lambs was recorded at birth (L0), and then on days 15 (L15) and 40 (L40). Redox biomarkers [total antioxidant capacity (TAC), glutathione (GSH), thiobarbituric acid reactive substances (TBARS)] were assayed in blood samples collected from lambs on days L0, L1, L2, L5, L10 and L40. Chemical analysis and antioxidant capacity were evaluated in colostrum and milk samples collected at the same time points with blood samples. Cytokines (IL-1β, IL-6, IL-10, IFN-γ) and immunoglobulin (IgG) were assayed in blood and colostrum samples collected from ewes on days L0 and L1, and in lambs’ blood on days L0, L1 and L2. The results revealed that body weight gain of newborn lambs did not differ between the two groups (p &gt, 0.05). Better redox status was found in MEL lambs until L2, as well as higher antioxidant capacity in the colostrum of MEL ewes compared to CON ones on day L0 (p &lt, 0.05). In MEL ewes’ colostrum, higher protein content was measured on day L0 and higher fat content on L1 compared to CON group (p &lt, 0.05). The highest level of IL-6 was found in MEL ewes on L1, with a concomitant increase of IL-10 level in MEL lambs in comparison to CON lambs on L2. Moreover, CON colostrum resulted in a higher level of IL-10 within time, coupled with an increased level of IgG found in lambs’ plasma on L2 (p = 0.04). This study indicated that melatonin could be administered as antioxidant and immune-modulatory regime in prenatally stressed offspring in order to cope with the crucial first days of their life. This effect of melatonin was also amplified by crosstalk between IL-6, IL-10 and IgG production, resulting in an improved quality of produced milk.

Food preparations, especially those based on animal products, are often accused of being responsible for the increase in food-borne infections, contributing to increased pressure on healthcare systems. The risk assessment in agri-food supply chains is of utmost importance for the food industry and for policymakers. A wrong perception of risks may alter the functioning of supply chains, thus, efforts should be devoted to communicating risks in an efficient way. We adopt a multidisciplinary approach to investigate how consumers perceive different food risks. Our analysis shows that planning effective communication strategies is very much important for efficiently informing consumers on food risks. We also comment on potential innovative ways to better organise the supply chains.

The present study was undertaken to evaluate the effect of protein fractions from bovine, caprine, and ovine milk on production of cytokines and reactive oxygen species (ROS) and reactive nitrogen species (RNS) by cultured peripheral blood mononuclear cells (PMBC) from infants with generalized epilepsy. Bovine, caprine, and ovine bulk milks were pasteurized and analyzed for chemical composition. Then, PBMC were isolated from 10 patients with generalized epilepsy (5 males; mean age 33.6±5.4mo). Production of tumor necrosis factor-α (TNF-α), IL-10, IL-6, and IL-1β was studied in cultured PBMC (from infants with epilepsy and controls) stimulated by bovine, caprine, and ovine milk and casein and whey protein fractions, and levels of ROS and RNS were measured in the culture supernatant. The ability of PBMC to secrete cytokines in response to milk and protein fraction stimulation may predict the secretion of soluble factor TNF-α in the bloodstream of challenged patients. Bovine, caprine, and ovine bulk milks induced low-level production of IL-10 by cultured PBMC in at least 50% of cases; the same behavior was observed in both casein and whey protein fractions for all species studied. Bovine and ovine milk and their casein fractions induced production of lower levels of IL-1β in 80% of patients, whereas caprine milk and its casein fraction induced the highest levels in 80% of patients. The amount of IL-6 detected after stimulation of PBMC by milk and its fractions for all species was lower than that of other proinflammatory cytokines. In the bovine, total free radicals were higher in bulk milk and lower in the casein fraction, whereas the whey protein fraction showed an intermediate level; in caprine, ROS/RNS levels were not different among milk fractions, whereas ovine had higher levels for bulk milk and casein than the whey protein fraction. Lower levels of ROS/RNS detected in PBMC cultured with caprine milk fraction could be responsible for the lower levels of TNF-α cytokine in the corresponding fraction. Cytokines might be useful biomarkers to discriminate the effects of foods on the inflammatory response; dietary strategies could help in alleviating the negative effects of epilepsy in infants.

The present research was undertaken to evaluate the effect of stage of lactation on immune competence of the goat mammary gland. The prevalence and etiology of intramammary infections and cytokine pattern were studied in individual caprine milk during lactation. A total of 1,200 caprine milk samples were collected during early, mid, and late lactation (less than 40 d, from 110 to 130 d, and more than 200 d in lactation, respectively) and analyzed for milk quality, bacterial cell load, and cytokine pattern. Contagious and environmental pathogens associated with intramammary infection were isolated from caprine milk. Nutritional components of caprine milk were affected by stage of lactation; fat, protein, casein, and whey protein increased along with lactation due to the lowest milk yield. Lactose content showed an opposite trend, with a reduced percentage passing from early to late lactation. The impaired rheological properties detected in early and late lactation milk were associated with a modification of milk quality. An impairment of microbial quality was detected at the end of lactation as a consequence of a major susceptibility of mammary gland to infections. The percentage of bacteriologically positive caprine milk samples was 44% throughout lactation. The incidence of bacteria associated with intramammary infection was different according to the stage of lactation with a major prevalence of Staphylococcus aureus, Enterobacteriaceae, Staphylococcus hyicus, and Streptococcus uberis in early lactation milk, Staphylococcus chromogenes, Streptococcus bovis, and Staphylococcus epidermidis in mid lactation, and Pseudomonas aeruginosa and Staphylococcus caprae in late lactation milk. The higher values of IL-1β, tumor necrosis factor-α, and IL-6 were ascribed to the major prevalence of Staphylococcus aureus at the beginning of lactation, whereas a higher level of IL-10 was detected in mid-lactation caprine milk and correlated with Staphylococcus chromogenes. Monitoring the cytokine pattern in milk helps to understand the immune status of the mammary gland, its ability to cope with bacteria, and the evolution of infections.

The purpose of this study was to evaluate the effects of different sources of polyunsaturated fatty acids (PUFA) supplemented in the diet on the welfare of lactating sheep exposed to high ambient temperature. The experiment was conducted during summer, it involved 32 ewes divided into 4 groups, and lasted 6 weeks. All groups were fed twice daily and received 1.8 kg/ewe/d of oat hay. The control group also received 1 kg/ewe/d of pelleted concentrate, whereas ewes in the experimental groups were supplemented with whole flaxseed (FS), seaweed Ascophyllum nodosum (Tasco®, AG), or their combination (FS + AG). At the beginning of the experiment, and weekly thereafter, the respiration rate, rectal temperature, body condition score (BCS) and body weight of the ewes were recorded. Milk samples were collected from each ewe weekly for the determination of the chemical composition, somatic cell count, and coagulating properties. Milk samples were also analyzed for their concentration of plasmin. Flaxseed diet during heat stress resulted in a reduction of the respiration rate, and in an increase of both milk and fat yield. The PUFA in the combined diet influenced the rectal temperature, and the respiration rate; also the milk yield and the concentration of plasmin (PL) increased in FS + AG diet. The lowest value of BCS recorded in ewes receiving the AG diet together with a rise in the respiration rate and rectal temperature suggested poor sheep welfare. Furthermore, sheep receiving the AG diet showed a consistent increase in the milk PL concentration that was responsible of the increase of milk clotting time and rate of clot formation.

Simple Summary Stressful conditions can play an important role in affecting welfare, productive performances and meat quality of lambs. The relation between nutrition and immune response has been investigated in the livestock production, particularly in dairy cattle and sheep. Due to costs related to animal feed it is useful to evaluate the proper feeding strategy supplementation for improving animal welfare and lamb meat quality. The present study aimed to evaluate the effects of supplementation with linseed, quinoa seed and their combination on metabolic profile, immune system, and cortisol response in blood and on meat quality of merinos derived lambs. Both linseed and quinoa supplementation enhanced the immune responses of lambs, and showed a hypo cholesterol effect on blood of lambs. Moreover, lambs supplemented with linseed resulted in the lowest level of cortisol secretion during the loading test demonstrating the link between stress and the immune system. In addition, data from the present experiment highlighted that linseed supplementation in lambs enhance meat quality producing a better meat tenderness. These findings should be considered for development of specific strategies aimed at improving the quality of meat and sustaining lambs’ welfare. Abstract In the last years several studies have investigated the strong relation between nutrition and immune response in the livestock production, particularly in dairy cattle and sheep. The aim of this study was to evaluate the effects of supplementation based on linseed, quinoa seeds and their combination on welfare, productivity and quality of meat from merinos derived lambs. 32 weaned lambs were divided into 4 experimental groups: quinoa (Q), linseed (LS) and combination of quinoa and linseed (LS + Q) that received the respective supplementation and control group (C) without supplementation. Lambs from all supplemented groups showed lower plasma urea, creatinine and cholesterol than control. Both linseed and quinoa supplementation enhanced the cell-mediated immune responses of lambs, furthermore, linseed supplementation resulted in the lowest level of cortisol secretion after handling, loading and transport. Meat from lambs supplemented with linseed and LS + Q showed the highest pH, at 1 and 3 h post-mortem, while, meat from all supplemented groups was more tender than meat from control. Results indicated that linseed and quinoa seeds supplementation can help the animal to cope with stressful events due to the close link between stress responses and the immune system and for improving meat quality in terms of better tenderness.

The objective of this study was to determine the effect of cooling upon calving in alleviating the adverse effects of heat stress in Holstein lactating cows. Production performance, indicators of metabolic status, immune response, and biomarkers of oxidative stress were measured. Based on mature equivalent milk production, parity, and calving date, 46 multiparous lactating cows were allotted to groups of equal sizes (n = 23); heat stressed (HS; BW = 658 ± 28 kg [mean ± SD]; BCS = 2.7 ± 0.18; parity = 3 ± 0.12) and cooled (CL; BW = 668 ± 23 kg; BCS = 2.8 ± 0.14; parity = 3 ± 0.25). Cows were housed in sand-bedded individual stalls equipped with misters and fans which were on from 1000 to 1800 hours for CL group. DMI and milk yield were measured from calving for 7 wk. Body condition score and BW were recorded weekly. Blood samples were collected weekly to measure the metabolic and antioxidant status, inflammatory cytokines, and immunoglobulins. Rectal temperature was measured daily at 1400 hour. Mean daily maximum temperature, minimum relative humidity, and maximum temperature–humidity index was 37.0 °C, 31.9%, and 83.4 for HS and 27.3 °C, 44.9%, and 75.7 for CL, respectively. Heat-stressed cows exhibited greater rectal temperature (39.8 vs. 39.1 °C) and lower feed intake (19.8 vs. 21.3 kg/d) relative to CL cows. Milk yield, including raw (31.2 vs. 38.6 kg/d) and fat- and protein-corrected (32.1 vs. 35.7 kg/d) milk, was lower in HS vs. CL cows, respectively. The percentages of milk protein (3.25 vs. 3.06), lactose (4.73 vs. 4.58), and solids-not-fat (8.63 vs. 8.38) but not milk fat (4.31 vs. 3.59) were higher in HS cows than in CL cows, respectively. Somatic cell score was greater in HS cows as compared with CL cows. Cooled cows lost less body condition as compared with HS cows. Blood plasma concentrations of glucose, non-esterified fatty acids, and β-hydroxybutyric acid were lower in HS cows. Blood plasma concentrations of malondialdehyde (2.13 vs. 1.84 nmol/mL), reactive oxygen species (579 vs. 561 U/mL), and total antioxidant capacity (4.49 vs. 4.06 U/mL) were greater in HS cows than in CL cows. Blood plasma concentrations of the inflammatory cytokines (tumor necrosis factor-α, interleukin-1α, and interleukin-2) and immunoglobulins (IgA, IgM, and IgG) were lower in HS cows than in CL cows. These findings demonstrated that cooling dairy cows during the early postpartum improved the production performance, indicators of metabolic status, immune response, and antioxidant capacity.

The post partum period is characterized by immunosuppression and increased disease susceptibility. Both phytosterols from microalga Dunaniella tertiolecta and dietary supplementation with n-3 polyunsaturated fatty acids (PUFA) influence cell proliferation and cytokine release during inflammation. The objective of this paper was the evaluation of the effects of physterols, extracted and purified from D. tertiolecta, on the in vitro immune responses of ewes supplemented with flaxseed during post partum. Twenty Comisana parturient ewes were divided in two balanced groups, and supplemented with flaxseed (FS, 250 g/day) or fed with a conventional diet (CON). Blood samples (15 mL) were collected for five weeks, starting from lambing, in order to isolate peripheral blood mononuclear cells (PBMC). Stimulated PBMC were treated with a total sterols fraction from D. tertiolecta (TS), a mix of ergosterol and 7-dehydroporiferasterol (purified extract, PE), and a mix of acetylated ergosterol and 7-dehydroporiferasterol (acetylated purified extract, AcPE), extracted and purified from D. tertiolecta at two concentrations (0.4 and 0.8 mg/mL). Results of the experiment demonstrated that n-3 PUFA from flaxseed induced an anti-inflammatory cytokine profile, with an increase of both IL-10, IL-6 and a decrease of IL-1β. TS, PE, and AcPE purified from D. tertiolecta showed an anti-proliferative effect on sheep PBMC regardless their chemical composition and concentration.

The experiment was carried out to evaluate the effects of a moderate level of flaxseed administration on milk coagulation properties and fatty acid profile of milk from two different breeds. The experiment was performed on 20 Italian Friesian cows and 20 Jersey cows divided into 2 groups of 10 animals each. The experimental diets were (1) a traditional diet (CON) administrated as unifeed and no supplemental fat and (2) a diet supplemented with 0·5 kg/d of whole flaxseed (FS). Cows were milked twice daily and milk yield was recorded. Milk samples were analysed at 1, 15, and 30 d of the experiment for composition, pH, and milk coagulation properties. To verify the effects of flaxseed administration on the coagulation properties of milk from Friesian and Jersey cows, an electrophoresis study on casein fractions was performed. Milk fatty acid profile can be improved by administrating a moderate level of flaxseed in the diet, however, milk fatty acid profile from Friesian and Jersey cows showed different contents of C18 : 1trans-11, SFA and MUFA. The results demonstrated that milk coagulating ability can be increased by flaxseed administration in both breeds as a result of different aggregation of casein micelles.

Sheep are considered one of the most resistant species with respect to climate change and high environmental temperatures. Most studies have focused on the effects of heat stress on the physiological mechanisms of adaptation in dairy cows; few studies have dealt with dairy sheep physiology and, above all, with sheep immunological responses. This chapter will focus on the complex network of mechanisms activated by heat stress as affecting immune responses in sheep. In particular, heat stress will be discussed as an attempt to animals’ homeostasis, thus affecting immune system and inducing activation of inflammatory processes. In particular, it will present a synopsis of the knowledge of the sheep immune system, and the changes induced by heat stress on both innate and adaptive immunity. Recent data suggesting a role of heat stress on perturbation of oxidative balance and the consequent effects on the immune system will be discussed. Finally, a discussion on the effects of heat stress on sheep mammary immunity will be presented.

The objective of this experiment was to study the effects of the unsaponified fraction (UP), the acetylated unsaponified fraction (AUP), and the total lipid fraction (TL) extracted and purified from Chlorella sorokiniana (CS) on the proliferation and cytokine profile of sheep peripheral blood mononuclear cells (PBMCs). Cells were cultured with 0.4 mg/mL and 0.8 mg/mL concentrations of each extract (UP, AUP, and TL fractions) and activated with 5 &mu, g/mL concanavalin A (ConA) and 1 &mu, g/mL lipopolysaccharide (LPS) at 37 &deg, C for 24 h. PBMCs cultured with ConA and LPS represented the stimulated cells (SC), and PBMCs without ConA and LPS represented the unstimulated cells (USC). Cell-free supernatants were collected to determine IL-10, IL-1&beta, and IL-6 secretions, on cells, measurement of proliferation was performed. All the extracts tested significantly decreased the cell proliferation, in particular, the UP fraction at 0.4 mg/mL showed the lowest proliferative response. Furthermore, at 0.8 mg/mL, the UP fraction enhanced IL-10 secretion. On the contrary, the TL fraction at 0.4 mg/mL induced an increase in IL-10, IL-6, and, to a lesser extent, IL-1&beta, secretions by cells. The AUP fraction did not change cytokine secretion. The results demonstrated that CS extracts could be useful ingredients in animal feed in order to minimize the use of antibiotics by modulating cell proliferation and cytokine response.

A large number of studies recognize glucocorticoids (Gc) as suppressors of inflammation; Gc exert an important role in coordinating the magnitude and duration of host immune responses. In the present in vitro investigation, we tested incremental levels of cortisol to verify the immunosuppressive or immunopermissive role of cortisol in sheep peripheral blood mononuclear cells (PBMC) after acute and chronic stress. Phytohemagglutinin (PHA)-stimulated PBMC were cultured for 24h and 96h at 37°C with 5% of CO 2 and varying cortisol levels: 10 ng/mL (baseline), 100 ng/mL (physiological poststressor), and 1,000 ng/mL [hyperactivated hypothalamic-pituitary-adrenal (HPA) axis]. The cell-free supernatants were collected for determination of IL-6, IL-1β, and IL-10 by ELISA, and the bromodeoxyuridine assay was performed on cells. Physiological cortisol concentration negatively affected the levels of IL-6 secreted by PBMC, resulting in increased cell proliferation after acute stress (24h of incubation). However, physiological cortisol concentration exhibited a reduction in cell proliferation induced by increased levels of IL-6 secreted by PBMC during chronic stress (96h of incubation). The cortisol concentration representing a hyperactivated HPA axis led to a reduction in cell proliferation after acute stress, which was probably induced by the elevated IL-10 production. Our results demonstrate that in sheep the effect of Gc on the immune system was related to the magnitude and the duration of stress. In particular, cortisol levels higher than physiological concentrations suppressed cell proliferation soon after acute stress. Instead, the physiological poststressor concentration of cortisol affected the immune responses in a bidirectional manner depending on the duration of the stressor.

The objective of the study was to characterize the immune profile of dairy ewes fed flaxseed, rich in polyunsaturated fatty acids (PUFA), around parturition. The hypothesis to be verified was that a physiological stressor, such as parturition, could be overcome with a nutritional manipulation in the diet of the animal in order to guarantee welfare of animals and to sustain their immune responses. Twenty Comisana ewes were divided in two groups (10 ewes/group), and fed a supplementation of whole flaxseed in the diet (FS group) or no supplementation (CON group). Blood samples were collected at parturition and then 7, 14, 21, 28, and 42 day post partum. Plasma samples were used to assess the humoral immune response after ovalbumin (OVA) immunization. At parturition, at 14 day, and 42 day post partum the level of plasma cytokines was assessed. The sheep showed a reduced responsiveness to OVA immunization. In FS ewes the IL-6 level remained unchanged until 14 day post partum and then significantly decreased from 14 day to 42 day post partum. IL-10 level was significantly higher in FS ewes than in CON ewes at 14 day. At parturition IL-1β level was significantly lower in FS ewes than in CON ewes and significantly decreased in both groups from parturition to 42 day. In conclusion, PUFA from flaxseed, as supplement in the diet of ewes around parturition can modulate sheep immune reactivity by influencing cytokine production.