Glycans are not necessary to maintain the pathobiological features of bovine spongiform encephalopathy.

The role of the glycosylation status of PrPC in the conversion to its pathological counterpart and on cross-species transmission of prion strains has been widely discussed. Here, we assessed the effect on strain characteristics of bovine spongiform encephalopathy (BSE) isolates with different transmission histories upon propagation on a model expressing a non-glycosylated human PrPC. Bovine, ovine and porcine-passaged BSE, and variant Creutzfeldt-Jakob disease (vCJD) isolates were used as seeds/inocula in both in vitro and in vivo propagation assays using the non-glycosylated human PrPC-expressing mouse model (TgNN6h). After protein misfolding cyclic amplification (PMCA), all isolates maintained the biochemical characteristics of BSE. On bioassay, all PMCA-propagated BSE prions were readily transmitted to TgNN6h mice, in agreement with our previous in vitro results. TgNN6h mice reproduced the characteristic neuropathological and biochemical hallmarks of BSE, suggesting that the absence of glycans did not alter the pathobiological features of BSE prions. Moreover, back-passage of TgNN6h-adapted BSE prions to BoTg110 mice recovered the full BSE phenotype, confirming that the glycosylation of human PrPC is not essential for the preservation of the human transmission barrier for BSE prions or for the maintenance of BSE strain properties.

Genistein effect in cultured ovine ovarian granulosa cells.

Genistein, an isoflavone has the potential to mimic, augment, or dysregulate the steroid hormone production pathways. We hypothesized that genistein affects the granulosa cell (GCs) functions through a series of biochemical, molecular, and genomic cascades. The present study was conducted to evaluate the impact of genistein exposure on GCs viability, apoptosis, and steroidogenesis. The present study involved 3/5 days of exposure to genistein on GCs collected from abattoir-derived ovine ovaries at doses of 0, 1, 10, 25, 50, and 100 µM. The harvested GCs were used for growth, cytotoxicity, and gene expression studies related to apoptosis, growth, and steroidogenesis. We observed that genistein had both stimulatory at 10 and 25 µM levels as well as inhibitory effects at 50 and 100 µM levels on the growth and proliferation of GCs. Genistein significantly decreased the levels of 17ß-estradiol at higher exposure (50 and 100 µM), whereas the progesterone level increased significantly as the genistein exposure increased. Additionally, genistein could also alter the mRNA expression of the steroidogenic receptor, enzymes, proteins, and growth-related genes suggesting that genistein could potentially alter the steroidogenic pathways. We conclude that genistein can interfere with cell survival and steroidogenesis by exhibiting a dose-dependent biphasic response on the viability, growth-related parameters, and the synthesis of 17ß-estradiol in the cultured GCs.

Analysis of fecal microbiome and metabolome changes in goats with pregnant toxemia.

BACKGROUND: Pregnancy toxemia is a common disease, which occurs in older does that are pregnant with multiple lambs in the third trimester. Most of the sick goats die within a few days, which can seriously impact the economic benefits of goat breeding enterprises. The disease is believed to be caused by malnutrition, stress, and other factors, that lead to the disorder of lipid metabolism, resulting in increased ketone content, ketosis, ketonuria, and neurological symptoms. However, the changes in gut microbes and their metabolism in this disease are still unclear. The objective of this experiment was to evaluate the effect of toxemia of pregnancy on the fecal microbiome and metabolomics of does. RESULTS: Eight pregnant does suspected of having toxemia of pregnancy (PT group) and eight healthy does during the same pregnancy (NC group) were selected. Clinical symptoms and pathological changes at necropsy were observed, and liver tissue samples were collected for pathological sections. Jugular venous blood was collected before morning feeding to detect biochemical indexes. Autopsy revealed that the liver of the pregnancy toxemia goat was enlarged and earthy yellow, and the biochemical results showed that the serum levels of aspartate aminotransferase (AST) and ß-hydroxybutyric acid (B-HB) in the PT group were significantly increased, while calcium (Ca) levels were significantly reduced. Sections showed extensive vacuoles in liver tissue sections. The microbiome analysis found that the richness and diversity of the PT microbiota were significantly reduced. Metabolomic analysis showed that 125 differential metabolites were screened in positive ion mode and enriched in 12 metabolic pathways. In negative ion mode, 100 differential metabolites were screened and enriched in 7 metabolic pathways. CONCLUSIONS: Evidence has shown that the occurrence of pregnancy toxemia is related to gut microbiota, and further studies are needed to investigate its pathogenesis and provide research basis for future preventive measures of this disease.

MFSD2A frameshift variant in Kerry Hill sheep with microcephaly.

Microcephaly is a rare neurodevelopmental disorder characterized by reduced skull circumference and brain volume that occurs sporadically in farm animals. We investigated an early-onset neurodegenerative disorder observed in seven lambs of purebred Kerry Hill sheep. Clinical signs included inability to stand or severe ataxia, convulsions, and early death. Diagnostic imaging and brain necropsy confirmed microcephaly. The pedigree of the lambs suggested monogenic autosomal recessive inheritance. We sequenced the genome of one affected lamb, and comparison with 115 control genomes revealed a single private protein-changing variant. This frameshift variant, MFSD2A: c.285dupA, p.(Asp96fs*9), represents a 1-bp duplication predicted to truncate 80% of the open reading frame. MFSD2A is a transmembrane protein that is essential for maintaining blood-brain barrier homeostasis and plays a key role in regulating brain lipogenesis. Human MFSD2A pathogenic variants are associated with a neurodevelopmental disorder with progressive microcephaly, spasticity, and brain imaging abnormalities (NEDMISBA, OMIM 616486). Here we present evidence for the occurrence of a recessively inherited form of microcephaly in sheep due to a loss-of-function variant in MFSD2A (OMIA 002371-9940). To the best of our knowledge, this is the first report of a spontaneous MFSD2A variant in domestic animals.

Characterization of TNSALP expression, localization, and activity in ovine utero-placental tissues†.

Tissue-nonspecific alkaline phosphatase (TNSALP; encoded by ALPL gene) has a critical role in the regulation of phosphate homeostasis postnatally. However, the utero-placental expression of TNSALP and the role in phosphate transport in pregnancy is poorly understood. Estrous cycles of ewes were synchronized, and ewes were euthanized and hysterectomized on Days 1, 9, or 14 of the estrous cycle or bred to fertile rams and euthanized and hysterectomized on Days 9, 12, 17, 30, 50, 70, 90, 110, or 125 of pregnancy. The expression of ALPL mRNA, immunolocalization of TNSALP protein, and quantification and localization of TNSALP enzymatic activity was performed on ovine endometria and placentomes. Day of the estrous cycle did not alter ALPL mRNA expression or enzymatic activity of TNSALP. TNSALP protein localized to uterine epithelial and stromal cells, blood vessels, myometrium, caruncular, and cotyledonary stroma. TNSALP activity was localized to uterine epithelia, blood vessels, caruncular stroma (from Day 70 of gestation), and the apical surface of chorionic epithelia (from Day 50 of gestation). TNSALP protein and activity localized to the apical surface of uterine epithelia during the estrous cycle and in early pregnancy. Endometrial TNSALP enzymatic activity was downregulated on Days 17 and 30 of gestation (P < 0.05). Expression of ALPL mRNA decreased in late gestation in endometria and placentomes (P < 0.05). TNSALP activity peaked in placentomes on Days 70 and 90 of gestation. Collectively, these results suggest a potential role of TNSALP in the regulation of phosphate transport and homeostasis at the maternal-conceptus interface in ruminants.

The ovine conceptus utilizes extracellular serine, glucose, and fructose to generate formate via the one carbon metabolism pathway.

Highly proliferative cells rely on one carbon (1C) metabolism for production of formate required for synthesis of purines and thymidine for nucleic acid synthesis. This study was to determine if extracellular serine and/or glucose and fructose contribute the production of formate in ovine conceptuses. Suffolk ewes (n = 8) were synchronized to estrus, bred to fertile rams, and conceptuses were collected on Day 17 of gestation. Conceptuses were either snap frozen in liquid nitrogen (n = 3) or placed in culture in medium (n = 5) containing either: 1) 4 mM D-glucose + 2 mM [U-13C]serine; 2) 6 mM glycine + 4 mM D-glucose + 2 mM [U-13C]serine; 3) 4 mM D-fructose + 2 mM [U-13C]serine; 4) 6 mM glycine + 4 mM D-fructose + 2 mM [U-13C]serine; 5) 4 mM D-glucose + 4 mM D-fructose + 2 mM [U-13C]serine; or 6) 6 mM glycine + 4 mM D-glucose + 4 mM D-fructose + 2 mM [U-13C]serine. After 2 h incubation, conceptuses in their respective culture medium were homogenized and the supernatant analyzed for 12C- and 13C-formate by gas chromatography and amino acids by high performance liquid chromatography. Ovine conceptuses produced both 13C- and 12C-formate, indicating that the [U-13C]serine, glucose, and fructose were utilized to generate formate, respectively. Greater amounts of 12C-formate than 13C-formate were produced, indicating that the ovine conceptus utilized more glucose and fructose than serine to produce formate. This study is the first to demonstrate that both 1C metabolism and serinogenesis are active metabolic pathways in ovine conceptuses during the peri-implantation period of pregnancy, and that hexose sugars are the preferred substrate for generating formate required for nucleotide synthesis for proliferating trophectoderm cells.

Characterization of microtubule-associated protein tau isoforms and Alzheimer's disease-like pathology in normal sheep (Ovis aries): relevance to their potential as a model of Alzheimer's disease.

Alzheimer's disease is a chronic neurodegenerative disease that accounts for up to 80% of all dementias. Characterised by deteriorations of memory and cognitive function, the key neuropathological features are accumulations of ß-amyloid and hyperphosphorylated tau, as 'plaques' and 'tangles', respectively. Despite extensive study, however, the exact mechanism underlying aggregate formation in Alzheimer's disease remains elusive, as does the contribution of these aggregates to disease progression. Importantly, a recent evaluation of current Alzheimer's disease animal models suggested that rodent models are not able to fully recapitulate the pathological intricacies of the disease as it occurs in humans. Therefore, increasing attention is being paid to species that might make good alternatives to rodents for studying the molecular pathology of Alzheimer's disease. The sheep (Ovis aries) is one such species, although to date, there have been few molecular studies relating to Alzheimer's disease in sheep. Here, we investigated the Alzheimer's disease relevant histopathological characteristics of 22 sheep, using anti-ß-amyloid (Abcam 12267 and mOC64) and phosphorylation specific anti-tau (AT8 and S396) antibodies. We identified numerous intraneuronal aggregates of both ß-amyloid and tau that are consistent with early Alzheimer's disease-like pathology. We confirmed the expression of two 3-repeat (1N3R, 2N3R) and two 4-repeat (1N4R, 2N4R) tau isoforms in the ovine brain, which result from the alternative splicing of two tau exons. Finally, we investigated the phosphorylation status of the serine396 residue in 30 sheep, and report that the phosphorylation of this residue begins in sheep aged as young as 2 years. Together, these data show that sheep exhibit naturally occurring ß-amyloid and tau pathologies, that reflect those that occur in the early stages of Alzheimer's disease. This is an important step towards the validation of the sheep as a feasible large animal species in which to model Alzheimer's disease.

Feed restriction around parturition does not affect colostrum immunoglobulin G concentration in dairy fat-tailed sheep but does affect performance and blood metabolites in newborn lambs.

This study aimed to investigate the effect of prepartum and postpartum feed restriction of fat-tailed dairy sheep on colostrum IgG concentration, and performance and blood metabolites of newborn fat-tailed lambs. Twenty fat-tailed dairy sheep were randomly allocated into control (Ctrl; n = 10) and feed restriction (FR; n = 10) groups. The Ctrl group received a diet that met 100% of energy requirements, both prepartum (from wk -5 to parturition) and postpartum (from parturition to wk 5). The FR group received a diet equivalent to 100, 50, 65, 80, and 100% of the energy requirements in wk -5, -4, -3, -2, and -1 relative to parturition, respectively. After parturition, the FR group received a diet equivalent to the 100, 50, 65, 80, and 100% of the energy requirements in wk 1, 2, 3, 4, and 5, respectively. At birth, lambs were assigned to their dam's experimental group. Both the Ctrl lambs (n = 10) and the FR lambs (n = 10) were allowed to suck colostrum and milk from the dams. Colostrum samples (50 mL) were collected at parturition (0 h) and then at 1, 12, 24, 36, 48, and 72 h postpartum. Blood samples were collected from all lambs before suckling colostrum (0 h) and then at 1, 12, 24, 36, 48, and 72 h after birth and weekly until the end of the experimental period (i.e., wk 5 relative to birth). The data were evaluated using the MIXED procedure of SAS (SAS Institute Inc.). The model included feed restriction, time, and the interaction feed restriction × time as fixed effects. The individual lamb was set as a repeated subject. Variables measured in colostrum and plasma were considered dependent variables, and significance was set at P < 0.05. Prepartum and postpartum feed restriction in fat-tailed dairy sheep did not affect colostrum IgG concentration. Consequently, no differences in blood IgG concentrations were observed in the lambs. In addition, the prepartum and postpartum feed restriction experienced by fat-tailed dairy sheep caused decreased body weight and milk intake in lambs from the FR group compared with the Ctrl group. Feed restriction also promoted increased concentration of blood metabolites such as triglycerides and urea in FR lambs compared with control lambs. In conclusion, prepartum and postpartum feed restriction in fat-tailed dairy sheep did not affect either colostrum IgG concentration or blood IgG concentration of the lambs. However, prepartum and postpartum feed restriction decreased lamb milk intake and, therefore, lamb body weight gain during the first 5 wk after birth.

In vitro addition of epinephrine, norepinephrine, and carnitine alters palmitate oxidation and esterification in isolated ovine hepatocytes.

Hepatocytes from 4 wethers were used to study the effects of carnitine and increasing concentrations of epinephrine and norepinephrine on palmitate oxidation and esterification. Liver cells were isolated from the wethers and incubated in Krebs-Ringer bicarbonate buffer with 1 mM [14C]-palmitate. Radiolabel incorporation was measured in CO2, acid-soluble products, and esterified products, including triglyceride, diglyceride, and cholesterol esters. Carnitine increased production of CO2 and acid-soluble products from palmitate by 41% and 216%, respectively, but had no effect on conversion of palmitate to esterified products. Epinephrine had a quadratic-increasing effect on palmitate oxidation to CO2, but norepinephrine did not increase palmitate oxidation to CO2. Neither epinephrine nor norepinephrine affected the production of acid-soluble products from palmitate. Increasing concentrations of norepinephrine and epinephrine linearly increased rates of triglyceride formation from palmitate. Increasing norepinephrine concentrations linearly increased diglyceride and cholesterol ester formation from palmitate in the presence of carnitine; epinephrine did not affect diglyceride or cholesterol ester formation. In general, catecholamine treatment had the greatest effect on the formation of esterified products from palmitate, and effects of norepinephrine were more pronounced than epinephrine. Conditions that result in catecholamine release might lead to fat accumulation in the liver.

Hematological parameters, antioxidant status, and gene expression of γ-INF and IL-1ß in vaccinated lambs fed different type of lipids.

This study was aimed to evaluate the effects of vegetable oils as calcium salt on immune responses and the expression of immune-related genes in vaccinated lambs. Twenty-four lambs (35 kg body weight, 6 months old) were assigned to four treatments with six replicates in a completely randomized design for 40 days. Four concentrates were formulated in which the calcium salts of palm oil, canola oil, corn oil, and flaxseed oil were used. On day 30 of the experiment, lambs were vaccinated by a dose of foot-and-mouth disease virus. The blood samples were collected from jugular vein 10 days after vaccination. The level of malondialdehyde and the activity of liver enzymes were the highest in lambs receiving corn oil and the lowest in lambs receiving flaxseed oil. The highest lymphocytes and the lowest neutrophil percentages were observed in lambs receiving flaxseed oil. There was a significant difference among treatments for the relative genes expression. Flaxseed oil significantly upregulated interferon-γ and corn oil upregulated interleukin-1ß. The highest titer against foot-and-mouth disease virus was related to lambs receiving flaxseed oil, and the lowest titer was related to lambs that received corn oil. Flaxseed oil had more beneficial effects on immune response than other oils.

The effect of dietary inclusion of Artemisia sieberi leaves on growth performance, feeding behaviors, ruminal fermentation, feed digestibility, and blood hemato-biochemical profile of growing male lambs.

The objective of this study was to investigate the impacts of replacing alfalfa hay with Artemisia sieberi leaves (ASL) on growth performance, feeding behaviors, total tract feed digestibility, ruminal fermentation as well as blood cells, and biochemical parameters. A total of 21 Kermani male lambs (average body weight (BW) of 25.2 ± 0.9 kg) were randomly divided into three groups and offered the following treatments: (1) basal diet (65% concentrate and 35% alfalfa hay as forage source), (2) ASL inclusion at 25% of forage source (8.75% of TMR, ASL25), and (3) ASL addition at 50% of forage source (17.5% of TMR, ASL50). The experiment lasted for 64 days, including 14 days of adaptation and 50 days of data collection. Dietary ASL inclusion decreased lambs' feed consumption (6.7 and 2.8% reduction in ASL25 and ASL50, respectively, p < 0.05) without affecting water intake, final BW, daily weight gain, and feed conversion ratio. Moreover, feeding behaviors, including time spent for eating, ruminating, and chewing activities, were uninfluenced by the dietary ASL addition on days 25 and 50 of the experiment. Based on the data provided, the substitution of alfalfa hay with ASL resulted in an increase in the digestibility of dry matter, organic matter, and neutral detergent fiber (p < 0.05). In addition, the ruminal fluid of ASL-fed lambs had a lower molar proportion of acetate and a higher proportion of propionate compared to that of the control animals (p < 0.05), whereas other volatile fatty acids (VFA) and total VFA were not different between the experimental groups. Blood hematocrit, hemoglobin, red cells, white cells, and their differentiation, platelets, glucose, total protein, albumin, globulin, triglyceride, calcium, phosphorus, aspartate aminotransferase, and alanine aminotransferase, urea, total antioxidant capacity, and malondialdehyde remained unchanged when ASL was included in the diet. Regardless of the slight decrease in dry matter intake, the lack of adverse effects of dietary ASL addition on growth performance, feeding behaviors, rumen fermentation, and blood hemato-biochemical parameters, along with positive impacts of ASL on propionate production and total tract feed digestibility, suggest that Artemisia sieberi leaves can be considered as a potential alternative feed resource for small ruminants.

Antioxidant action of yerba mate on carcass and meat characteristics and fatty acid profile in meat and fat of lambs finished in tropical pastures.

The objective was to evaluate the effect of the supplementation with natural antioxidants from Yerba mate (Ilex paraguariensis St. Hil.) on the performance, carcass and meat characteristics, and fatty acid profile in meat and fat of Texel × Ile de France lambs kept in Brachiaria-grass pasture. Twenty lambs with a mean age of 90 days and 26.3 kg of BW were distributed in a completely randomized design in two treatments with 10 replicates/treatment: Control-protein-energy supplement based on corn and soybean meal (150 g/kg DM of crude protein) without the addition of natural antioxidant yerba mate and Yerba Mate - energy protein supplement with the addition of yerba mate (110 g/kg DM). There was no treatment effect on animal performance and carcass characteristics. The subcutaneous fat thickness was significantly higher for lambs treated without yerba mate. There was no effect of nutritional treatments on the oxidative markers of lamb meat. The muscle of the animals treated with yerba mate showed a higher concentration of unsaturated and polyunsaturated fatty acids elaidic, linolenic, thymnodonic, docosadienoic, and ceorvonic. The fat of the animals treated with yerba mate showed a higher concentration of palmitoleic acid, linoelaidic acid, heneicosanoic acid, dihomo-γ-linolenic acid, and arachidonic acid. The concentration of polyunsaturated fatty acids was significantly higher in meat and fat of lambs treated with yerba mate. Including yerba mate in the diet of lambs kept on Brachiaria pastures improves the lipid profile of polyunsaturated fatty acids in meat and fat, without altering the performance of the animals.

Comparison of the effect of Saccharomyces cerevisiae-Megasphaera elsdenii and buffer on growth performance, digestibility, ruminal histomorphometry, and carcass characteristics of fattening lambs in high concentrate diet.

This study aimed to investigate the effect of rumen pH-adjusting additives in the high-concentrated diet on functional traits, nutrient digestion, some meat parameters, and histomorphometry, and rumen histopathology. Twenty-four Arabia male lambs with 3 to 4 months old and initial body weight of 23.9 ± 3.15 kg were used in a completely randomized design with three treatments and eight replicates. The study was 77 days, including 14 days of the adaptation period and 63 days of the record taking and sampling period. The experimental treatments consisted of a control diet, control diet + sodium bicarbonate buffer, control diet + Megasphaera elsdenii, and Saccharomyces cerevisiae (bacterial-yeast). Rumen fluid was taken by stomach tube at 3 h after morning feeding to measure pH. The lambs were weighed every 3 weeks during the period, and the body weight changes, average daily gain, and total weight gain were measured, and the feed conversion ratio was calculated. At the end of the experiment, the lambs were slaughtered, and the longissimus dorsi muscle was prepared to determine the meat parameters. For histological studies, the abdominal rumen sac was sampled. There were no differences among treatments in dry matter intake (DMI), daily weight gain (ADG), and feed conversion ratio (P > 0.05). Propionate concentration was higher in the bacteria-yeast treatment than other treatments (P < 0.05). Protein digestibility was higher in control and bacteria-yeast treatments than buffer treatment (P < 0.05). The percentage of meat protein, carcass weight, and dressing percentage in bacterial-yeast treatment was higher than other treatments (P < 0.05). Rumen wall thickness in the buffer and bacterial-yeast receiving treatments was greater than the control treatment and was significant in the buffer treatment compared to the control treatment (P < 0.05). The thickness of rumen epithelial tissue in the buffer and bacterial-yeast recipient treatments was less than the control treatment (P < 0.05). Rumen papillae thickness was higher in the control treatment than other treatments (P < 0.05). Hydropic degeneration and parakeratosis were less in pH-regulating treatments than in control. The results showed that the use of Megasphaera elsdenii could be an effective way to modulate the ruminal fermentation conditions of lambs fed with high concentrate diets. In addition, to increaseing dressing percentage and meat protein, it can also reduce tissue damage and improve ruminal tissue structure.

Replacing ground Rhodes grass hay with soyhulls in the pelleted diet: effects on ingestive behavior, nutrient digestibility, blood metabolites, growth performance, and economic viability of intensive fattening Lohi lambs.

This study aimed to evaluate the effects of replacing soyhulls (SH) for ground Rhodes grass hay (RGH) in a pelleted diet on ingestive behavior, digestibility, blood metabolites, growth performance, and economic viability of fattening Lohi lambs. A total of thirty male lambs (age: 5 months; body weight 20.4 ± 0.24 kg) were allotted to one of the three diets (n = 10 lambs/diet) under a completely randomized design. The diets were with 25% RGH inclusion (control), replacing 15% RGH with 15% SH as a fiber source (SH-15), and containing only 25% SH inclusion on a dry basis (SH-25). Ingestive behavior parameters like time spent (min/day), bouts frequency (number/day), and bout length (min/bout) for feeding, drinking, rumination, chewing, standing, and lying were not influenced (P > 0.05) by replacing RGH with SH. The dry matter (DM) and neutral detergent fiber (NDF) chewing rate, rumination rate, and feeding efficiency were also not changed (P > 0.05) by dietary treatments, whereas total dry matter and NDF intakes, and their rumination efficiencies were lower (P < 0.05) for the SH-25 than for the rest of the treatments. The digestibility of NDF and acid detergent fiber (ADF) was higher (P < 0.05) for the SH-25 than for the rest of the treatments. The concentrations of blood metabolites like glucose, blood nitrogen, and cholesterol were not changed (P > 0.05) among the treatment groups. Similarly, performance parameters like dry matter intake, initial and final body weight, average daily gain, and feed conversion ratio were similar (P > 0.05) across the treatments. However, the incidence of loose fecal consistency was higher (P < 0.05) for SH-25 than for the control. The values of economic efficiency were better for SH-25-fed lambs than those fed the rest of the treatments. Based on the results, substituting SH for RGH in a pelleted diet improved the digestibility of fiber fractions, and economics without affecting the growth performance, and blood metabolites of fattening lambs. However, lower rumination efficiency and loos fecal consistency are evidencing less effectiveness of SH fiber.

Folic acid supplementation during pregnancy modulates hepatic methyl metabolism and genes expression profile of neonatal lambs of different litter sizes.

Maternal folic acid (FA) plays an important role in the fetus development, but it is unknown the response of hepatic metabolism in the offspring from different litter sizes to maternal FA supplementation. In the present study, this was done by feeding the ewes with 0, 16 and 32 mg/(kg·DM) FA supplemented diet during pregnancy and analysing the hepatic one-carbon metabolism-related indices and gene expression in the neonatal lambs of different litter sizes (twins, TW; triplets, TR). Regardless of litter sizes, the concentrations of folate, methionine, S-adenosylmethionine and DNA methyltransferase increased significantly, but homocysteine and S-adenosylhomocysteine decreased in the liver of newborn lambs from ewes whose diet was supplemented with FA. In TW, maternal FA status has little effect on hepatic genes expression profile of newborn lambs, and no significant enriched pathway was found. However, DEG involved in cell proliferation such as CCNA2, CCNB2, CCNE2, CDK1 and BUB1 were significantly enriched when the ewes were supplemented with FA in TR groups. In addition, nucleotide synthesis-related genes such as POLD1, POLD2, MCM4 and MCM5 were enriched markedly in DNA replication and pyrimidine metabolism pathways in triplets when a higher FA ingestion [32 mg/(kg·DM)] was implemented in ewes. This finding demonstrated that the hepatic methyl metabolism in TW and TR newborn lambs was regulated by maternal FA status. The hepatic cell proliferation and nucleotide metabolism related genes in TR were more susceptible to maternal dietary FA supplementation during pregnancy.

Effect of L-citrulline supplementation on sperm characteristics and hormonal and antioxidant levels in blood and seminal plasma of rams.

With the aim of providing a theoretical basis for the application of L-citrulline (L-Cit) in animal husbandry, the effects of L-Cit on reproductive hormone levels, antioxidant capacity and semen quality of rams were studied by feeding them varying doses of L-Cit. A total of 32 rams were randomly divided into four groups with eight rams each. After all rams were trained to donate sperm normally, the control group was fed a basic diet, whereas the experimental groups I, II and III were provided with feed supplemented with 4, 8 and 12 g/d of L-Cit respectively. The experiment was conducted for 70 days, during which blood samples were collected from the jugular vein on days 0, 15, 30, 45 and 60, and semen samples were collected on days 0, 20, 40 and 60. In the same group, 100 µl of semen was used to test for quality, The rest of the semen sample and blood samples were centrifuged at 600 g for 15 min, and the supernatant and serum, respectively, were used to determine the levels reproductive hormones and antioxidant indices. Ram semen samples were also collected on day 70 and used to study sperm plasma membrane, substitution and mitochondrial membrane potential. Compared with the control group, the groups receiving L-Cit showed an increase in sperm concentration and number of linear motile sperm (p < .01); a decrease in the number of dead sperm (p < .01); an increase in sperm viability, particularly in groups II and III (p < .01); and an increase in sperm mitochondrial membrane potential (p < .01). Moreover, groups I, II and III showed significantly higher levels of serum gonadotropin-releasing hormone (GnRH), glutathione peroxidase (GSH-Px) and nitric oxide (NO) (p < .01). Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels increased in groups I (p < .05), II (p < .05) and III (p < .01), whereas testosterone (T), catalase (CAT) and superoxide dismutase (SOD) levels increased in groups I and II (p < .01). Serum total antioxidant capacity (T-A) increased (p < .05), whereas both hydroxyl radical (·OH) and peroxy radical ( O 2 · - ) levels decreased (p < .01). Compared with the control, all groups had significantly higher SOD and GSH-Px in their seminal plasma (p < .01), and groups I, II (p < .05 for both) and III (p < .01) had higher levels of GnRH and FSH. LH, CAT and NO levels increased in group I (p < .05), II and III (p < .01 for both); malondialdehyde levels decreased in groups I, II (p < .05 for both) and group III (p < .01); and O 2 · - levels decreased in groups I, II and III (p < .01). Under our experimental conditions, GnRH, FSH, LH, T, CAT, SOD, T-A, GSH-PX and NO levels in the serum and seminal plasma of rams receiving L-Cit increased, whereas Oestradiol (E2 ), O 2 · - and ·OH levels in the seminal plasma decreased; this improved the semen quality of rams supplemented with L-Cit. Moreover, supplementation with 12 g/d gave the best results.

Improving the functional value of meat quality by feeding with protected fat supplementation in Morkaraman lambs.

This study investigated the effect of feeding different levels of protected fat (calcium soaps of tallow) on fattening performance, carcass characteristics, meat quality, fatty acid composition of Longissimus Thoracis (LT) muscle and economic evaluation of 45 fat-tailed Morkaraman male lambs. The lambs were randomly divided into three groups including a control diet without protected fat (CON), supplemented with 2% protected fat (group PF2), and supplemented with 4% protected fat (group PF4) during the fattening period. Eight lambs randomly selected from each group were slaughtered to determine carcass characteristics and meat quality. Supplementing protected fat significantly improved the fattening performance and carcass characteristics (p<0.05). The groups were similar in terms of pH, colour, ether extract, dry matter and sensory evaluations, except for the crude protein of group PF2. Myristic acid and stearic acid (p<0.05), which are saturated fatty acids (SFA), were decreased with the supplementation of protected fat, while palmitic acid was increased (p<0.01). The fattening with protected fat had a highly significant positive effect on important unsaturated fatty acids such as oleic acid (p<0.01) and resulted in an increase of approximately 91% in the conjugated linoleic acid (CLA) (p<0.05). SFA was significantly decreased, and health indicators were improved (p<0.01). In conclusion, the supplementation of protected fat had a positive effect on fattening performance, carcass traits, chemical, sensory properties and economic evaluation.

Effects of using siris (Albizia lebbeck) foliage in the diet of fattening lambs on nutrient digestibility, blood and rumen parameters, growth performance, and meat quality characteristics.

This study aimed to investigate the effect of substitution of siris foliage with alfalfa forage in the diet of fattening lambs on digestibility, fermentation, and growth performance of fattening lambs. In the present experiment, 27 8-month-old Arabi lambs (31.3 ± 6) with an initial weight of 28.8 ± 1.99 kg were used in a completely randomized design. The effect of experimental diets on dry matter intake was not significant; however, the diets had a significant effect on the intake of neutral detergent fiber (NDF), acid detergent fiber (ADF), and crude protein (P < 0.05). The effect of experimental diets on the apparent digestibility of dry matter, organic matter, NDF, ADF, and crude protein was not significant (P < 0.05). Ammonia nitrogen concentration, pH, and a total population of ruminal fluid protozoa and blood parameters were not affected by experimental diets. Parameters of fattening performance such as feed intake, live weight changes, feed conversion ratio, some carcass traits such as mean weight and size of carcass parts, and colorimetric indices of muscle tissue in the order of fattening lambs were not affected by experimental diets. The use of foliage of siris in the diet of fattening lambs as a substitute with part of alfalfa had no adverse effect on the characteristics studied in the present experiment. Therefore, siris be recommended as part of the diet of fattening lambs.

The effects of feeding chickpea grains on the lactating performance and blood metabolites of ewes.

An experiment was conducted to see how replacing soybean meal with chickpea grains (CHPE) affected the performance, milk production, digestibility, nitrogen (N) balance, and blood metabolites of Awassi ewes. Thirty Awassi ewes with an initial body weight of 53.2 ± 2.31 kg and aged between 4 and 5 years were selected and randomly assigned to different feeding treatments (10 ewes per diet). Dietary dry matter (DM) was (1) 0 g/kg CHPE (CON); (2) 150 g/kg CHPE (CHPE150); and (3) 300 g/kg CHPE (CHPE300). The costs of these diets were determined using current ingredient prices. The experiment lasted 9 weeks. During the trial, ewes and their lambs were kept in individual pens. Daily nutrient intake was monitored. Every 2 weeks, the body weight (BW), milk output, and composition of the ewes were measured. In comparison to the CON diet, adding the CHPE to the experimental diets lowered feed costs (US$) per ton by 11% for CHPE150 and 21% for CHPE300. The CHPE300 group had a lower (P < 0.05) consumption of neutral detergent fiber (NDF) than the CHPE150 group, while the CON group had the highest (P < 0.05) intake of NDF. The intake of ether extract (EE) (g/d) was higher (P < 0.05) in the ewes fed the CHPE300 compared to the ewes fed CHPE150, and it was lowest in ewes fed the CON diet. When the ewes were fed the CHPE300 and the CHPE150 diets, the total solids (g/kg) in each kg of milk produced per day was higher (P < 0.05) than when they were fed the CON diet. The cost per kg of milk produced by the ewes was lowered (P < 0.05) when the CHPE was added to their diets. When the ewes on the CHPE300 and the CHPE150 diets were compared to the ewes on the CON diet, crude protein (CP) digestibility was higher (P < 0.05). The EE digestibility was higher (P < 0.05) for the CHPE300 group, while the CHPE150 group had an intermediate outcome. The CHPE300 group had lower amounts of N in their feces and urine (P < 0.05) than the other groups. The retention of N (g/100 g) in the ewes fed the CHPE300, and the CHPE150 diets was higher (P < 0.05) than in the ewes fed the CON diet. As a result, the current research demonstrates that part of soybean meal and barley grain could be replaced with chickpeas in the diets of lactating Awassi ewes.

Insights into the lipidome and primary metabolome of the uterus from day 14 cyclic and pregnant sheep†.

In ruminants, conceptus elongation requires the endometrium and its secretions. The amino acid, carbohydrate, and protein composition of the uterine lumen during early pregnancy has been defined in sheep; however, a comprehensive understanding of metabolomic changes in the uterine lumen is lacking, particularly with respect to lipids. Here, the lipidome and primary metabolome of the uterine lumen, endometrium, and/or conceptus was determined on day 14 of the estrous cycle and pregnancy. Lipid droplets and select triglycerides were depleted in the endometrium of pregnant ewes. In contrast, select ceramides, diglycerides, and non-esterified fatty acids as well as several phospholipid classes (phosphatidylcholine, phosphatidylinositol, phosphatidylglycerols, and diacylglycerols) were elevated in the uterine lumen of pregnant ewes. Lipidomic analysis of the conceptus revealed that triglycerides are particularly abundant within the conceptus. Primary metabolite analyses found elevated amino acids, carbohydrates, and energy substrates, among others, in the uterine lumen of pregnant ewes. Collectively, this study supports the hypothesis that lipids are important components of the uterine lumen that govern conceptus elongation and growth during early pregnancy.