Rumen fermentation and epithelial gene expression responses to diet ingredients designed to differ in ruminally degradable protein and fiber supplies.

Although numerous studies exist relating ruminal volatile fatty acid (VFA) concentrations to diet composition and animal performance, minimal information is available describing how VFA dynamics respond to diets within the context of the whole rumen environment. The objective of this study was to characterize how protein and fiber sources affect dry matter intake, rumen pH, fluid dynamics, fermentation parameters, and epithelial gene expression. Four diet treatments (soybean meal or heat-treated soybean meal and beet pulp or timothy hay) were delivered to 10 wethers. The soybean meals served as crude protein (CP) sources while the beet pulp and timothy hay represented neutral detergent fiber (NDF) sources. Feed intake, rumen pH, fluid pool size, and fluid passage rate were unaffected by treatment. Butyrate synthesis and absorption were greater on the beet pulp treatment whereas synthesis and absorption of other VFA remained unchanged. Both CP and NDF treatment effects were associated with numerous VFA interconversions. Expression levels of rumen epithelial genes were not altered by diet treatment. These results indicate that rumen VFA dynamics are altered by changes in dietary sources of nutrients but that intake, rumen environmental parameters, and the rumen epithelium may be less responsive to such changes.

Feeding wheat dried distillers' grains with solubles increases conjugated linoleic acid and unsaturated lipids in ovine milk without adversely affecting milk yield.

The aim of this research communication was to examine the effect of dietary supplementation with wheat-based dried distillers' grains with solubles (DDGS), a by-product of bioethanol production, on yield, composition, and fatty acid (FA) profile of ewe milk. Forty-five purebred mid-lactating Chios ewes (average milk yield 2.23 kg/d in 96 ± 5 d in lactation) were offered three iso-nitrogenous and iso-energetic diets (15 animals per diet) for a 10 d adaptation period followed by a 5-week recording and sampling period. The diets contained 0, 6, and 12% DDGS on DM basis for the DG0, DG6, and DG12 treatment, respectively, as a replacement of concentrate mix, whilst concentrate-to-forage ratio remained at 60:40 in all treatments. Individual milk yield, milk composition, and FA profile were recorded weekly and analyzed using a complete randomized design with repeated measurements. No significant differences were observed among groups concerning dry matter intake (overall mean of 2.59 kg/d), milk yield or 6% fat-corrected milk and milk protein percentage or protein yield. Milk fat percentage was decreased in the DG12 (4.76%) compared to DG0 (5.69%) without, however, significantly affecting the daily output of milk fat. The concentration of all major saturated FA between C4:0 to C16:0 was reduced, whereas long-chain (>16 carbons), mono-unsaturated and poly-unsaturated FAs were increased in the milk of DDGS groups. Among individual FA, increments of oleic acid and C18:1 trans-monoenes like C18:1 trans-10 and C18:1 trans-11 were demonstrated in DG12 group, whereas linoleic and conjugated linoleic acid (CLA cis-9, trans-11) were elevated in both DDGS groups compared to control. Changes in FA profile resulted in a decline in the atherogenic index of milk by 20% and 35% in DG6 and DG12 treatments, respectively, compared with control. In conclusion, feeding DDGS to dairy ewes increased the levels of unsaturated FA that are potentially beneficial for human health without adversely affecting milk, protein or fat yield.

Gallic acid promotes the in vitro development of sheep secondary isolated follicles involving the phosphatidylinositol 3-kinase pathway.

This study was conducted to evaluate the effect of addition of gallic acid as the single antioxidant to the base medium for in vitro culture of sheep secondary follicles and if the phosphatidylinositol 3-kinase (PI3K) pathway is involved in the action of gallic acid. Secondary follicles were isolated and cultured for 12 days in α-MEM supplemented with bovine serum albumin (BSA), insulin, glutamine, hypoxanthine, transferrin, selenium, and ascorbic acid (control medium: α-MEM+) or in α-MEM supplemented with BSA, insulin, glutamine, hypoxanthine and different concentrations of gallic acid (25, 50 or 100 µM), thus replacing transferrin, selenium and ascorbic acid in the medium. Follicle morphology, glutathione (GSH), and mitochondrial activity, and meiotic resumption were evaluated. Furthermore, inhibition of PI3K pathway was performed by pretreatment with LY294002. After 12 days of culture, the follicle survival in a medium containing 100 µM gallic acid was similar (P > 0.05) to α-MEM+ and greater (P < 0.05) compared with other gallic acid concentrations. Antrum formation, follicle diameter, GSH, and mitochondrial activity, and meiotic resumption, however, were greater (P < 0.05) when 100 µM gallic acid was included in the α-MEM+ culture medium compared with the control medium. Furthermore, LY294002 inhibited (P < 0.05) follicle survival, development, and meiotic resumption stimulated by 100 µM gallic acid. In conclusion, concentration of 100 µM of gallic acid can be a substitute for transferrin, selenium, and ascorbic acid in the base medium during in vitro culture of sheep secondary follicles, inducing follicle development likely through the PI3K pathway.

Periconceptional nutrition with spineless cactus (Opuntia ficus-indica) improves metabolomic profiles and pregnancy outcomes in sheep.

We tested whether periconceptional nutrition with cladodes from the cactus, Opuntia ficus-indica, with or without protein-enrichment, improved the metabolomic profile and reproductive outcomes of adult female sheep. Sixty Rambouillet ewes of similar body weight were randomly allocated among three nutritional treatments that were fed during the breeding period (34 days): Control (Control; n = 20), Opuntia (Opuntia; n = 20) and protein-enriched Opuntia (E-Opuntia; n = 20). There were no effects of treatment on body weight but assessment of urine samples indicated that, for 76 metabolites, the Control and Opuntia groups differed completely (P < 0.05), whereas there was overlap between the Control and E-Opuntia groups. It appears that, in Opuntia-fed and Control-fed sheep, different functional groups are activated leading to changes in the metabolism of glucose, tyrosine, methane, and glycerolipids. Fertility and reproductive rate tended to be higher in the Opuntia (70% and 95%) and E-Opuntia (90% and 110%) groups than in the Control (55% and 65%), and an orthogonal contrast revealed the difference between Control and Opuntia to be significant for both reproductive variables (P < 0.05). We conclude that nutritional supplementation with Opuntia cladodes, with or without protein enrichment, increased fertility rate and reproductive rate of female sheep, without any accompanying increases in body weight. Our observations suggest that the reproductive responses to Opuntia do not simply reflect a response to good nutrition, but might be caused by specific metabolites/metabolomic pathways, perhaps by an activation of the metabolism of glucose, methane, tyrosine and glycerolipids. There are few reports relating these metabolomic compounds with the metabolism of the sheep, let alone with reproductive efficiency. The novelty of these discoveries suggests that we need further research into the mechanisms through which nutrition affects the reproductive system.

Supplementing pregnant Merino ewes with caffeine to improve neonatal lamb thermoregulation and viability.

Percentages of lamb mortalities prior to weaning are a significant and persistent problem for the Australian sheep industry. Maternal caffeine supplementation reduced stillbirths and improved viability in piglets; however, the efficacy of caffeine in enhancing viability and lamb survival is equivocal. The aim of this study was to determine the optimal concentration and duration of maternal caffeine supplementation to improve lamb viability; time to stand and suck with consumption of colostrum occurring, as well as survival to weaning. Multiparous Merino ewes were supplemented with either 0 (CTL), 10 (C10) or 20 mg/kg (C20) body weight (BW) caffeine in feed after day 120 of gestation (dG), or 20 mg/kg BW (LC20) caffeine from dG 142 until parturition. Ewes were housed indoors in individual lambing pens from dG 130 to 72 -h post-partum (pp). Values for pp ewe and lamb variables were analysed using a generalised linear mixed model in IBM SPSS version 25. While ewes within the CTL, C10 and LC20 groups consumed more caffeine compared to C20 ewes (P =  0.001), lambs of C20 ewes had greater rectal temperatures at 20 -h pp (P =  0.021), greater 4 -h serum IgG concentrations (P =  0.041), a longer latency to first sucking bout (P =  0.030), and a greater number of sucking attempts (P =  0.044) compared to lambs from CTL, C10 and LC20 ewes. These results indicate that caffeine supplementation during late-gestation stimulates neonatal lambs as a result of increased sucking (4 -h serum IgG) and increased temperatures within the first 24 -h.

Beneficial effects of rumen-protected methionine on nitrogen-use efficiency, histological parameters, productivity and reproductive performance of ruminants.

Providing essential amounts of balanced nutrients is one of the most vital aspects of livestock production. Among nutrients, protein has an essential role in many physiological functions of animals. Amino acids in needs for both high and medium yielding ruminant animals are not fully covered by microbial degraded feed sources in the rumen of animals, and they must be met by protecting the proteins from being broken down in the rumen; hence, the dietary supplementation of rumen-protected proteins (RPP), including mainly rumen-protected methionine (RPM), became imperative. Many researchers are interested in studying the role of (RPM) in ruminant animals concerning its effect on milk yield, growth performance, digestibility, dry matter intake and nitrogen utilization efficiency. Unfortunately, results obtained from several investigations regarding RPM indicated great fluctuation between its useful and useless effects in ruminant nutrition particularly during early and late lactation period; therefore, this review article may be helpful for ruminant farm owners when they decide to supplement RPM in animal's diet. Conclusively, supplementation of RPM often has a balanced positive influence, without any reported negative impact on milk yield, growth performance and blood parameters especially in early lactating ruminant animals and when used with the low crude protein diet.

Neuroanatomical basis of the nerve growth factor ovulation-induction pathway in llamas†.

The objective of the study was to characterize the anatomical framework and sites of action of the nerve growth factor (NGF)-mediated ovulation-inducing system of llamas. The expression patterns of NGF and its receptors in the hypothalamus of llamas (n = 5) were examined using single and double immunohistochemistry/immunofluorescence. We also compare the expression pattern of the P75 receptor in the hypothalamus of llama and a spontaneous ovulator species (sheep, n = 5). Both NGF receptors (TrkA and P75) were highly expressed in the medial septum and diagonal band of Broca, and populations of TrkA cells were observed in the periventricular and dorsal hypothalamus. Unexpectedly, we found NGF immunoreactive cell bodies with widespread distribution in the hypothalamus but not in areas endowed with NGF receptors. The organum vasculosum of the lamina terminalis (OVLT) and the median eminence displayed immunoreactivity for P75. Double immunofluorescence using vimentin, a marker of tanycytes, confirmed that tanycytes were immunoreactive to P75 in the median eminence and in the OVLT. Additionally, tanycytes were in close association with GnRH and kisspeptin in the arcuate nucleus and median eminence of llamas. The choroid plexus of llamas contained TrkA and NGF immunoreactivity but no P75 immunoreactivity. Results of the present study demonstrate sites of action of NGF in the llama hypothalamus, providing support for the hypothesis of a central effect of NGF in the ovulation-inducing mechanism in llamas.

Preference for inorganic sources of calcium and phosphorus by sheep as a function of need.

We hypothesized that ruminants discriminate among different mineral supplements and modify their selection as a function of need. Forty lambs were offered four-way choices among P, Ca, and Mg sources: monosodium phosphate (MSP), Magnesium phosphate (MGP), Monocalcium phosphate (MCP), and Dicalcium phosphate (DCP); and two-way choices between DCP and the other mineral sources (Baseline). Subsequently, lambs were randomly assigned to four groups (10 lambs/group) and fed rations such that levels of Ca and P were low (LCa_LP), adequate (NCa_NP), low in Ca and adequate in P (LCa_NP), or adequate in Ca and low in P (NCa_LP). Four- and two-way choice tests were repeated (Post-deficiency). During Baseline, lambs ate and preferred MSP > MGP>DCP = MCP (p < .0001), showing that they discriminated among minerals. This pattern remained during four-way Post-deficiency tests, but lambs in LCa_LP, LCa_NP, and NCa_LP showed increased preference for MCP, MGP, and DCP, respectively, relative to other groups and Baseline (p < .05). Some mineral preference also changed in relation to serum concentration of P and Ca. Thus, lambs discriminated among minerals and some groups modified their preferences from Baseline to Post-deficiency choice tests, likely influenced by mineral needs, which in turn contributed to rectify mineral imbalances.

Carcass traits and meat quality of Texel lambs raised in Brachiaria pasture and feedlot systems.

Our hypothesis suggests an improvement in carcass and meat quality of pasture-finished animals by introducing concentrate feed into diets. This study aimed to evaluate the effect of different levels of protein-energy supplementation on carcass and meat characteristics of Texel lambs raised on Brachiaria pastures, and compare the results with those obtained from confined animals. Thirty 2-month-old intact lambs were divided into five treatments: pasture with mineral supplementation, pasture containing daily supplies of 0.8, 1.6, and 2.4% BW protein-energy supplementation, and confinement (reference treatment). All animals were slaughtered 104 days after the beginning of the experiment. Lambs submitted to 1.6% and 2.4% BW supplementation presented similar characteristics to animals kept in confinement and were superior to animals treated with 0 or 0.8% of BW supplementation levels (p < .05). Increases of subcutaneous fat thickness were verified for 0.0, 0.8, 1.6, 2.4% BW, and confinement treatments, being respectively of 0.25, 0.74, 1.61, 1.69, and 1.98. Conversely, treatments had no influence on meat physical-chemical characteristics, being all considered moderately soft, juicy (tender), and with mild tastes and odors. Lambs raised on Brachiaria pastures have high-quality meat, but more protein-energy supplementation is needed to be an alternative for the feedlots for young animals.

Moringa oleifera leaves extract enhances fresh and cryopreserved semen characters of Barki rams.

This study was carried out to evaluate the efficacy of Moringa oleifera leaves extract (MOLE) to improve the characters of fresh and cryopreserved semen of Barki rams compared to vitamin E and Selenium combination. Twenty-four mature Barki rams (50-70 Kg) were randomly assigned into three groups, eight rams each. The first group was given distilled water orally. The second group was given MOLE orally daily at a dose of 40 mg/kg. The third group was injected with a combination of vitamin E and selenium at a dose of 3 ml (4.5 mg sodium selenite and 204 mg vitamin E)/head i.m twice a week for 64 days. Moringa oleifera leaves extract increased semen volume, sperm concentration, activities of seminal plasma catalase, glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD), alkaline phosphatase (ALP), acid phosphatase (ACP), levels of ascorbic acid and total antioxidant capacity (TAC). In addition, it significantly increased post thawing sperms motility, viability index, membrane integrity, and the activities of post thawing semen antioxidant enzymes. While it decreased seminal plasma concentration of malondialdehyde (MDA) and acrosomal defects and DNA fragmentation of sperm in cryopreserved semen. Vitamin E and selenium decreased semin volume, sperm concentration, seminal plasma ascorbic acid, TAC concentrations and activities of antioxidant enzymes while it increased sperm abnormalities, DNA fragmentation and MDA concentration in seminal plasma. This study indicated that Moringa oleifera leaves extract improved the characters of the fresh and cryopreserved semen of Barki rams via improving seminal plasma antioxidant defense mechanism.

Impact of docusate and fauna-free on feed intake, ruminal flora and digestive enzyme activities of sheep.

Four Small-tail Han male hogget sheep, fitted with rumen cannula and fed the same basal diet were used to study the impacts of docusate (DOC) and fauna-free on the voluntary feed intake (VFI), and ruminal protozoal, bacterial and fungal counts and the digestive enzyme activities. By a 4 × 4 Latin square design, sheep were given no DOC (the control), 2 doses of DOC: 1.2 and 3.0 g/kg diet or oral dose of 6.0 g/d DOC for three days (fauna-free treatment) in each period of 18 days, the last three days of which were for sampling the rumen fluid. Compared with the control, 1.2 g/kg of DOC supplementation significantly resulted in increases of 18.0% VFI and 44% bacterial count, and no significant change in the fungal number. Supplementing DOC reduced protozoal number in a dose-dependent manner. The fibre degradation enzyme activity in rumen fluid increased by 17.7% with a concomitant 10% increase in volatile fatty acids (VFA); the protease activity was reduced by 23% with a corresponding reduction in rumen ammonia by 42%. In contrast, supplementing 3.0 g/kg of DOC has adverse effects on those measures compared with 1.2 g/kg of DOC. Defaunation was accompanied with substantial increases in the bacterial and fungal counts, but had no significant influences on VFI and the enzyme activity for starch, protein and pectin digestion, and small changes in fibre digestion enzymes and the total VFA compared with the control. A high correlation (r2  = 0.82) was noted between VFI and the total activity of fibre digestion enzymes and VFA. It was proposed that fibre digestion rate in the rumen is a primary factor for determining VFI in sheep, and dietary supplementation of 1.2 g/kg of DOC could partially result in enhanced activity of fibre digestive enzyme in the rumen and increase VFI.

Metagenomic and volatile profiles of ripened cheese obtained from dairy ewes fed a dietary hemp seed supplementation.

Chemical and organoleptic properties of dairy products largely depend on the action of microorganisms that tend to be selected in cheese during ripening in response to the availability of specific substrates. The aim of this work was to evaluate the effects of a diet enriched with hemp seeds on the microbiota composition of fresh and ripened cheese produced from milk of lactating ewes. Thirty-two half-bred ewes were involved in the study, in which half (control group) received a standard diet, and the other half (experimental group) took a diet enriched with 5% hemp seeds (on a DM basis) for 35 d. The dietary supplementation significantly increased the lactose in milk, but no variations in total fat, proteins, caseins, and urea were observed. Likewise, no changes in total fat, proteins, or ash were detected in the derived cheeses. The metagenomic approach was used to characterize the microbiota of raw milk and cheese. The phyla Proteobacteria and Firmicutes were in equally high abundance in both control and experimental raw milk samples, whereas Bacteroidetes was less abundant. The scenario changed when considering the dairy products. In all cheese samples, Firmicutes was clearly predominant, with Streptococcaceae being the most abundant family in the experimental group. The reduction of taxa observed during ripening was in accordance with the increment (relative abundance) of the starter culture Lactococcus lactis and Streptococcus thermophilus, which together dominate the microbial community. The analysis of the volatile profile in ripened cheeses led to the identification of 3 major classes of compounds: free fatty acids, ketones, and aldehydes, which indicate a prevalence of lipolysis compared with the other biochemical mechanisms that characterize the cheese ripening.

Dietary supplementation of brown seaweed (Sargassum latifolium) alleviates the environmental heat stress-induced toxicity in male Barki sheep (Ovis aries).

Heat stress (HS) is the most potent environmental stressors for livestock in tropical and subtropical regions. HS induced splanchnic tissue hypoxia and intestinal oxidative damage, leading to endotoxemia and systemic inflammation. The present study evaluated and compared the modulatory effects of feeding Barki male sheep (Ovis aries) on a standard concentrated diet containing 2% or 4% of the brown seaweed (Sargassum latifolium) followed by roughage for 40 consecutive days on the toxicity-induced by exposure to severe environmental HS (temperature-humidity index = 28.55 ± 1.62). The present study showed that the diet containing Sargassum latifolium (especially 4%) modulated significantly (P < 0.05-0.001) almost all changes shown in the HS-exposed sheep including the increase in the thermo-respiratory responses (skin and rectal temperatures, and respiration rate) and the resulted dyslipidemia, anemia, and systemic inflammation (blood leukocytosis, the elevation in the erythrocyte sedimentation rate, and the increase in serum proinflammatory cytokines and heat shock protein-70 concentrations). In addition, Sargassum latifolium improved significantly (P < 0.05-0.001) the body-weight gain, kidney functions (especially at the high dose), and blood antioxidant defense system (total antioxidant capacity, and the activities of catalase and superoxide dismutase) in the HS-exposed sheep, as well as protected the animals from oxidative tissue damage and the risk of atherosclerosis. In conclusion, feeding sheep with the diet containing 4% of Sargassum latifolium was safe and suitable for animal nutrition, as well as efficiently alleviated the harmful effects of the environmental HS in Barki sheep through improving the animal antioxidant defense system, and regulating the thermo-respiratory and inflammatory responses.

Maternal arginine supplementation enhances thermogenesis in the newborn lamb.

Body temperature maintenance is one of the most important physiological processes initiated after birth. Brown adipose tissue (BAT) is an essential mediator of thermogenesis in many species and is responsible for 50% of the heat generated in the newborn lamb. To determine if maternal arginine supplementation could enhance thermogenesis in the neonate, we randomly assigned 31 multiparous Suffolk ewes, gestating singletons or twins, to receive intravenous injections of either l-arginine (27 mg/kg body weight; n = 17) or sterile saline (n = 14) three times daily from day 75 to 125 of gestation (term = 147). Following parturition, lambs were removed from their mothers and subjected to 0 °C cold challenges at 4 and 22 h of age. Rectal temperatures were higher for the duration of the cold challenges in lambs from arginine-treated ewes compared with lambs from saline-treated ewes (P < 0.05). Elevated rectal temperatures were associated with increased (P < 0.05) circulating glycine and serine concentrations in lambs. The mRNA expression of genes related to BAT function changed over time, but not between lambs from arginine-treated vs. saline-treated ewes. Results indicate that maternal arginine treatment increases neonatal thermogenesis after birth. Although the underlying mechanisms remain to be elucidated, these data are a first step in improving neonatal survival in response to cold.

Exogenous melatonin and male foetuses improve the quality of sheep colostrum.

Colostrum is the first product secreted by the mammary gland to transfer immunity to the newborn, especially through immunoglobulins (Ig) G. Melatonin is an immunomodulatory factor and there is evidence that it has a direct effect on IgG production. To evaluate the effects of melatonin treatment during pregnancy, litter size and offspring sex on colostrum quality, sixty pregnant Rasa Aragonesa ewes were divided into three groups: one group received a melatonin implant at the third month of pregnancy (3M, n = 13), another group at the fourth month (4M, n = 18) and the remaining ewes were not implanted (Control, C, n = 29). Immediately after lambing, a sample of colostrum was collected and IgG, crude protein and fat content analysed. Timing of melatonin implantation (p < .001), and offspring sex (p < .01) had a significant effect on IgG concentration. Colostrum of treated ewes had a higher mean (±SEM) IgG concentration than that of the control ewes (55.54 ± 3.09 and 49.50 ± 4.36 mg/ml, respectively; p < .05), mainly because the concentration in the 4M group was significantly (p < .001) higher than it was in the other groups. The relationship between lamb sex and IgG (p < .01) and %CP (p < .05) was evident in singletons (ewes with a male lamb: 54.57 ± 5.37 mg IgG/ml, 15.42 ± 0.82%CP; ewes with a female lamb: 34.66 ± 4.30 mg/ml, 13.18 ± 0.73%CP). The presence of a female in the litter was associated with significantly (p < .01) lower colostrum IgG levels (litters with at least one female: 49.33 ± 3.42 mg/ml; litters with no females: 58.24 ± 4.00 mg/ml). Among 4M ewes, female foetuses had significantly (p < .01) lower IgG levels whether they carried singletons or multiple lambs. In conclusion, treatment with melatonin implants at the fourth month of pregnancy resulted in a higher colostrum quality based on IgG concentration.

Dietary nitrate and presence of protozoa increase nitrate and nitrite reduction in the rumen of sheep.

Nitrate ( NO 3 - ) supplementation is an effective methane (CH4 ) mitigation strategy for ruminants but may produce nitrite ( NO 2 - ) toxicity. It has been reported that rumen protozoa have greater ability for NO 3 - and NO 2 - reduction than bacteria. It was hypothesised that the absence of ruminal protozoa in sheep may lead to higher NO 2 - accumulation in the rumen and a higher blood methaemoglobin (MetHb) concentration. An in vivo experiment was conducted with defaunated (DEF) and faunated (FAU) sheep supplemented with 1.8% NO 3 - in DM. The effects of rumen protozoa on concentrations of plasma and ruminal NO 3 - and NO 2 - , blood MetHb, ruminal volatile fatty acid (VFA) and ruminal ammonia (NH3 ) were investigated. Subsequently, two in vitro experiments were conducted to determine the contribution of protozoa to NO 3 - and NO 2 - reduction rates in DEF and FAU whole rumen digesta (WRD) and its liquid (LIQ) and solid (SOL) fractions, incubated alone (CON), with the addition of NO 3 - or with the addition of NO 2 - . The results from the in vivo experiment showed no differences in total VFA concentrations, although ruminal NH3 was greater (p < .01) in FAU sheep. Ruminal NO 3 - , NO 2 - and plasma NO 2 - concentrations tended to increase (p < .10) 1.5 hr after feeding in FAU relative to DEF sheep. In vitro results showed that NO 3 - reduction to NH3 was stimulated (p < .01) by incoming NO 3 - in both DEF and FAU relative to CON digesta. However, adding NO 3 - increased (p < .05) the rate of NO 2 - accumulation in the SOL fraction of DEF relative to both fractions of FAU digesta. Results observed in vivo and in vitro suggest that NO 3 - and NO 2 - are more rapidly metabolised in the presence of rumen protozoa. Defaunated sheep may have an increased risk of NO 2 - poisoning due to NO 2 - accumulation in the rumen.

Effect of dry medicinal plants (wormwood, chamomile, fumitory and mallow) on in vitro ruminal antioxidant capacity and fermentation patterns of sheep.

The objective of this study was to determine the effect of dry medicinal plants (wormwood, chamomile, fumitory and mallow) and dietary substrates containing a mix of the plants on the end products of in vitro ruminal and intestinal fermentation, rumen protozoan population and ruminal antioxidant capacity of sheep. The experiment consisted of fermentations with the four plants used individually as the sole substrate and fermentation of a mix of medicinal plants (Plants): meadow hay:barley grain (MH:B), 700/300 w/w and Plants:MH:B, 100/600/300 w/w/w. The experiment was conducted using the in vitro gas production technique (IVGPT) with 35 ml of buffered inocula and approximately 250 mg (DM basis) of substrate incubated for 24 hr at 39°C in anaerobic conditions. Quantitative analyses of the bioactive compounds by ultra-high-resolution mass spectrometry in Plants identified three main groups: flavonoids (22 mg/g DM), phenolic acids (15 mg/g DM) and alkaloids (3 mg/g DM). The total antioxidant capacity (TAC) of the plant extracts and rumen fluid was analysed using a ferric reducing antioxidant power assay. The values of total and individual short-chain fatty acids, acetate:propionate ratio, pH and total gas production were significantly affected by the single plant substrates and inocula (p < .001). Apart from these parameters, the values of ammonia N, methane production and total gas produced were decreased in Plants:MH:B in comparison with MH:B (p < .001). A positive correlation was recorded between total polyphenols content and TAC of plant extracts (R2  = .778, p < .001). The counts of the total ciliate protozoan population in rumen fluid after 24 hr of fermentation were not significantly different (p > .05). Results suggest that the dietary substrate containing the medicinal plant mix possessed strong ruminal antioxidant capacity, had the potential to reduce methane emission and ammonia concentration and caused desirable changes in the gastrointestinal ecosystem.

Supplementation of by-products from grape, tomato and myrtle affects antioxidant status of dairy ewes and milk fatty acid profile.

The aim of this study was to evaluate the effect of diets containing different dried by-products on milk and blood plasma antioxidant capacity of dairy ewes. Thirty-six Sarda ewes were assigned to four treatments: control (CON; no by-product), 100 g/day of grape marc (GM), 100 g/day tomato pomace (TP) and 75 g/day of exhausted myrtle berries (EMBs). The superoxide dismutase (SOD), glutathione reductase (GR), glutathione transferase (GST) and glutathione peroxidase (GSH-Px) in blood, and SOD, GR and lactoperoxidase (LPO) in milk were determined. Total antioxidant capacity (FRAP and ABTS assays), malondialdehyde (MDA) and protein carbonyls (PCs) were also measured. Milk fatty acid profile was investigated by gas chromatography. The results showed higher antioxidant capacity measured by FRAP or ABTS assays and a reduction in MDA in GM plasma than CON. All by-products enhanced the protection of milk proteins by oxidation, as evidenced by lower values of PCs compared with CON. GM supplementation increased PUFAn-6, due to increase in C18:2n-6, the main component of GM compared with CON. All by-products did not modify the nutritional indexes of milk fat. In conclusion, dietary GM may enhance protection against oxidative condition of dairy ewes, whereas TP and EMB need further research to define the optimum inclusion level in sheep diet.

Partitioning of fatty acids into tissues and fluids from reproductive organs of ewes as affected by dietary phenolic extracts.

Evaluation of dietary interventions with regard to fertility problems often observed in ruminant livestock is of global interest. Though the effects of polyphenol supplementation in ruminants on digestion and food quality are well described, the impact on reproductive tissues and fluids remains scarcely investigated. These compounds protect dietary unsaturated fatty acids (FA) from oxidation and biohydrogenation and thus saturation. In addition, modification of the expression of genes associated with FA metabolism may occur. Therefore, we characterized for the first time the FA profiles of reproductive tissues and fluids and investigated their potential modification by dietary polyphenols in 22 cyclic ewes. The animals were randomly divided into four groups and fed a basal diet of meadow hay and one of four concentrate types either non-supplemented (control) or supplemented with grape seed extract, Acacia mearnsii bark extract (13 g/kg dry matter (DM) each) or a combination of both (26 g/kg DM). After 10 weeks of feeding, the animals were slaughtered. Samples of reproductive (oviduct, uterus) and metabolically differently active tissues (liver, muscle, adipose) as well as of plasma and fluids from oviduct and uterus were analysed for their FA composition. In addition, the expression of lipid metabolic and antioxidant genes was analysed in all tissues except the adipose tissue. Fatty acid profiles in tissues and fluids as well as gene expression in tissues significantly differed between the different fluids and tissues. In contrast, only a few diet and matrix (fluid or tissue) × diet interactions were observed. Still, the FA profile of the uterus was the only one not at all affected by the diet. The mRNA expression was not affected by the diet for most of the genes investigated, which might in part be explained by the similar plasma polyphenol concentrations found at slaughter. Overall, our findings contribute to an improved understanding of the characteristic FA composition of reproductive tissues and fluids in sheep. In addition, the effect of polyphenols on different tissues, fluids and tissue gene expression has been confirmed as described in other animal species.

Effects of oregano essential oil on in vitro ruminal fermentation, methane production, and ruminal microbial community.

Different inclusion rates of oregano essential oil (OEO) were investigated for their effects on ruminal in vitro fermentation parameters, total gas, methane production, and bacterial communities. Treatments were (1) control, 0 mg/L of OEO (CON); 13 mg/L (OEO1); 52 mg/L (OEO2); 91 mg/L (OEO3); and 130 mg/L (OEO4), each incubated with 150 mL of buffered rumen fluid and 1,200 mg of substrate for 24 h using the Ankom in vitro gas production system (Ankom Technology Corp., Fairport, NY). Treatment responses were statistically analyzed using polynomial contrasts. Digestibility of DM, NDF, and ADF increased quadratically with increasing OEO inclusion rates. Digestibility of DM and NDF were highest for OEO2, whereas ADF digestibility was highest for OEO3, compared with CON, with the remaining treatments being intermediate and similar. Ammonia nitrogen concentrations decreased from CON at a quadratic rate with increasing OEO inclusion rates, and OEO2 had the lowest concentration compared with the other groups. Total VFA, acetate, propionate, butyrate, valerate, and isovalerate concentrations linearly decreased with increasing OEO inclusion rates. Total gas production levels by CON and OEO4 were greater than those of OEO1, OEO2, and OEO3 in a quadratic response, and methane production linearly decreased from CON, compared with OEO4, at a decreasing rate with OEO inclusion rates. As determined by 16S rRNA sequencing, the α biodiversity of ruminal bacteria was similar among OEO inclusion rates. Increasing OEO inclusion rates linearly increased the relative abundance of Prevotella and Dialister bacteria. Several bacteria demonstrated different polynomial responses, whereas several bacteria were similar among increasing OEO inclusion rates. These results suggested that OEO supplementation can modify ruminal fermentation to alter VFA concentrations and reduce methane emissions by extensively altering the ruminal bacterial community, suggesting an optimal feeding rate for future animal studies of approximately 52 mg/L for mature ruminants.