Control Sheep Research Articles

Developmental programming: Transcriptional regulation of visceral and subcutaneous adipose by prenatal bisphenol-A in female sheep

BackgroundBisphenol-A (BPA) exposure is widespread and early life exposure is associated with metabolic syndrome. While visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) are implicated in the development of metabolic syndrome, the adipose depot-specific effects of prenatal BPA treatment are poorly understood. ObjectiveTo determine the impact of prenatal BPA exposure on genome-wide gene expression of VAT and SAT depots. MethodsRNA sequencing was performed on SAT and VAT from 21-month old control and prenatal BPA-treated female sheep. Gene expression and pathway differences between SAT and VAT depots with or without prenatal BPA-treatment and the effect of prenatal BPA treatment on each depot were tested. ResultsThere were 179 differentially expressed genes (padjusted < 0.05, log2-fold change >2.5) between SAT and VAT. Development and immune response pathways were upregulated in SAT, while metabolic pathways were upregulated in VAT. These adipose depot-specific genes and pathways were consistent with prenatal BPA-treatment. In SAT, BPA-treatment resulted in differential expression of 108 genes (78% upregulated with BPA) and altered pathways (immune response downregulated, RNA processing upregulated). In contrast in VAT, BPA-treatment differentially expressed 4 genes and upregulated chromatin and RNA processing pathways. ConclusionPrenatal BPA-treatment induces adult depot-specific alterations in RNA expression in inflammation, RNA processing, and chromatin pathways, reflecting the diverse roles of SAT and VAT in regulating lipid storage and insulin sensitivity. These adipose tissue transcriptional dysregulations may contribute to the metabolic disorders observed in prenatal BPA-treated female sheep.

Effects of fermented Caragana korshinskii on the intramuscular fat content and expression of FABP3, UBE3C, ADRB3, LIPE, and SCD in different muscles of Tan sheep

The aim of this study was to investigate the effects of fermented Caragana korshinskii on the intramuscular fat content and varied expression of the intramuscular fat deposition-related genes FABP3, UBE3C, ADRB3, LIPE, and SCD among four muscle tissues (m. psoas, gluteus, quadriceps, and supraspinatus) of Tan sheep. Twenty-eight male animals of similar age (270 ± 10 days) and weight (24.6 ± 1.06 kg) were randomly divided into a control group (fed the basal diet) and an experimental group (fed the same diet except 10% of corn stalks were replaced with fermented C. korshinskii). Soxhlet petroleum-ether extraction and quantitative real-time PCR were applied to evaluate the fat content and gene expression in tissues, respectively. We observed a significant improvement (P &amp;lt; 0.05) in the intramuscular fat contents in the m. gluteus and supraspinatus of treated sheep compared to those of non treated sheep. The FABP3 mRNA level was markedly higher (P &amp;lt; 0.05) in the m. quadriceps and supraspinatus of treated sheep than in the control sheep. UBE3C mRNA levels were significantly decreased in the m. gluteus, quadriceps, and supraspinatus (P &amp;lt; 0.05) of treated sheep compared with those of the control sheep. ADRB3 mRNA levels were significantly lower (P &amp;lt; 0.05) in the m. psoas, gluteus, and supraspinatus of sheep fed fermented C. korshinskii than in the control group, whereas LIPE mRNA levels were significantly increased (P &amp;lt; 0.05) in the m. gluteus and quadriceps of sheep fed fermented C. korshinskii. The SCD mRNA levels in m. psoas, quadriceps, and supraspinatus of sheep fed fermented C. korshinskii were significantly higher than those of the control group (P &amp;lt; 0.05). Our results indicated that fermented C. korshinskii could partially replace the roughage used in Tan sheep feed, and its substitution affected the intramuscular fat content and altered the expression of intramuscular fat deposition-related genes. The present study lays a solid foundation for further exploring the utilization of C. korshinskii in ruminant husbandry.

Effects of Nano-Copper Poisoning on Immune and Antioxidant Function in the Wumeng Semi-Fine Wool Sheep.

The Wumeng semi-fine wool sheep is vital to the production system of the Wumeng mountainous area in Southwest China. To study the effect of nano-copper poisoning in the Wumeng semi-fine wool sheep, nano-copper poisoning model was established in sheep. We determined blood physiological and biochemical parameters, immune indexes, and antioxidant indicators. The results showed that Hb, RBC, and PCV levels in blood from the nano-copper group were markedly lower (P < 0.01) than those in the control group. Serum LDH, AST, ALT, CPK, and Cp from the nano-copper group were significantly higher (P < 0.01) than those in control animals. Serum SOD, GSH-Px, CAT, and T-AOC from the nano-copper group were significantly lower (P < 0.01) than those in control sheep, and MDA content in serum from the nano-copper group were markedly higher (P < 0.01) than those in control sheep. The levels of IL-2, IL-6, IL-1β, IgA, IgM, and IgG from the nano-copper group were significantly lower (P < 0.01) than those in the control group. It was concluded that nano-copper poisoning could not only affect the immune function of Wumeng semi-fine wool sheep but also reduce the antioxidant capacity.

Histologic safety of transcranial focused ultrasound neuromodulation and magnetic resonance acoustic radiation force imaging in rhesus macaques and sheep

BackgroundNeuromodulation by transcranial focused ultrasound (FUS) offers the potential to non-invasively treat specific brain regions, with treatment location verified by magnetic resonance acoustic radiation force imaging (MR-ARFI). ObjectiveTo investigate the safety of these methods prior to widespread clinical use, we report histologic findings in two large animal models following FUS neuromodulation and MR-ARFI. MethodsTwo rhesus macaques and thirteen Dorset sheep were studied. FUS neuromodulation was targeted to the primary visual cortex in rhesus macaques and to subcortical locations, verified by MR-ARFI, in eleven sheep. Both rhesus macaques and five sheep received a single FUS session, whereas six sheep received repeated sessions three to six days apart. The remaining two control sheep did not receive ultrasound but otherwise underwent the same anesthetic and MRI procedures as the eleven experimental sheep. Hematoxylin and eosin-stained sections of brain tissue (harvested zero to eleven days following FUS) were evaluated for tissue damage at FUS and control locations as well as tissue within the path of the FUS beam. TUNEL staining was used to evaluate for the presence of apoptosis in sheep receiving high dose FUS. ResultsNo FUS-related pre-mortem histologic findings were observed in the rhesus macaques or in any of the examined sheep. Extravascular red blood cells (RBCs) were present within the meninges of all sheep, regardless of treatment group. Similarly, small aggregates of perivascular RBCs were rarely noted in non-target regions of neural parenchyma of FUS-treated (8/11) and untreated (2/2) sheep. However, no concurrent histologic abnormalities were observed, consistent with RBC extravasation occurring as post-mortem artifact following brain extraction. Sheep within the high dose FUS group were TUNEL-negative at the targeted site of FUS. ConclusionsThe absence of FUS-related histologic findings suggests that the neuromodulation and MR-ARFI protocols evaluated do not cause tissue damage.

Reduced Na+ K+ -ATPase activity may reduce amino acid uptake in intrauterine growth restricted fetal sheep muscle despite unchanged ex vivo amino acid transporter activity.

Fetuses with intrauterine growth restriction (IUGR) have reduced muscle mass that persists postnatally, which may contribute to their increased risk for adult onset metabolic diseases, such as diabetes and obesity. Amino acid transporter-mediated histidine uptake and system L amino acid transporter activity were similar in sarcolemmal membranes isolated from control and IUGR hindlimb skeletal muscle. Activity of Na+ K+ -ATPase, which is responsible for establishing the sodium gradient necessary for system A and N amino acid transporter function, was significantly reduced in IUGR skeletal muscle sarcolemma compared to control. ATP content was lower in IUGR skeletal muscle. Expression and phosphorylation of proteins in the mechanistic target of rapamycin pathway were similar in control and IUGR skeletal muscle homogenate. Our data suggest that lower Na+ K+ -ATPase activity, which reduces the driving force for active amino acid transport, and lower ATP availability contribute to reduced amino acid uptake and protein synthesis in IUGR fetal skeletal muscle. Fetuses with intrauterine growth restriction (IUGR) have lower muscle mass that persists postnatally. Using a sheep model of placental insufficiency and IUGR, we have previously demonstrated lower net total uptake of amino acids by the fetal hindlimb and lower skeletal muscle protein synthesis rates. To investigate the mechanisms underlying these changes, we tested the hypothesis that ex vivo amino acid transporter and Na+ K+ -ATPase activity is reduced, and ex vivo ATP levels are lower in hindlimb skeletal muscle of the IUGR fetus. We developed a novel protocol to measure transporter-mediated histidine uptake, system L amino acid transporter activity and Na+ K+ -ATPase activity using sarcolemmal membranes isolated from hindlimb muscle of control (CON, n=11-12) and IUGR (n=12) late gestation fetal sheep. We also determined ATP content and the activity of insulin and mechanistic target of rapamycin (mTOR) signalling, which are involved in regulating cellular amino acid uptake and protein synthesis, by measuring the expression and phosphorylation of AKT, 4E-BP1, eIF2α, AMPKα, p70 S6 kinase and rpS6 in muscle homogenates. Transporter-mediated histidine uptake and system L activity were similar in control and IUGR sarcolemma, although ex vivo Na+ K+ -ATPase activity was lower by 64% (P=0.019) in IUGR sarcolemma. ATP content was lower by 25% (P=0.007) in IUGR muscle. Insulin, AMPK, and mTOR signalling activity was similar in control and IUGR muscle. We speculate that reduced muscle sarcolemmal Na+ K+ -ATPase activity and lower ATP content diminishes the sodium gradient in vivo, resulting in a reduced driving force for sodium-dependent transporters and subsequently lower muscle amino acid uptake.

Pathogens of intestinal cysts of sheep, their epizootology, diagnosis and profilactics

The article provides information on literature and personal research on the causes and effects of intestinal cestodosis in sheep in Uzbekistan, their occurrence in all seasons and animals of different ages, their current epizootological status, the methods for their differentiation and prevention. However, this is not the case in lambs, which may be thought to be due to the fact that cement damage begins later than in adult sheep. In the control sheep, the separation of the cement joints was observed during the experiment. In sheep and lambs of this control group, further deworming was performed. In the case of worm infections, the gastrointestinal cestodosis of sheep is most prevalent in terms of its prevalence, economic damage, and low levels of study. Given the widespread use of worms in animals and the emergence of various diseases in farm animals and poultry, their study will undoubtedly provide an opportunity to address not only veterinary but also broader socio-economic issues. This is also partly due to the immediate release of potted flowers or other green plants to domestic animals in wet weather. Intermediate hosts of cestodes - oribatid mites are common in all climatic and geographical regions of Uzbekistan - irrigated plains, foothills and desert-pasture biocenoses. After the skeleton, the cervical portion begins and is of different length in the cestodes of different systematic groups: the neck is very short (2.5–3 mm to 5–10 mm) in the parasites of the generation of monies and the neck is longer (30–40 cm). and the Avitellina lineage is longer and longer.

Effect of subclinical mastitis on the yield and cheese-making properties of ewe’s milk

Abstract This review covers an update of scientific knowledge about productive and technological consequences of subclinical mastitis in sheep milk. The literature reports individual milk yield losses of 2.6–43.1 %, being modulated by several factors as infection severity, production level, causal agents, and unilateral or bilateral IMI (1 or 2 infected glands, respectively). A compensatory increase of milk production from the uninfected gland when only one half was infected has been quantified in 6.6 %, compared with healthy halves of control sheep. This compensatory adaptation highlights the risk of underestimating subclinical mastitis in sheep. The mammary gland response is quick and milk yield losses in absolute terms remained constant within the following weeks, both when infection appear during lactation and when it is present from lambing. With respect to the changes on main components in milk due to subclinical mastitis it has been clearly established a decrease in the concentration of lactose and an increase of that of whey proteins. The role played by lactose as an osmotic regulator results in a more accentuated decrease of its concentration in milk. This is why lactose is considered at present as a reliable potential indicator of subclinical mastitis. Whey proteins increase as a result of the increase of the blood-milk barrier permeability and the increased proteolysis of caseins. However, the content in milk of fat and casein are modified depending on the magnitude of milk yield reduction, being affected by a concentration or dilution effect. In any case, the ratio casein to protein (parameter independent of the milk volume) decreases as a result of infection. The impairment of physical and chemical characteristics due to decreased udder health status is the responsible of the negative effect of increased SCC on the coagulation properties of milk, the curd yield and the quality of cheese. Low ratio of casein to protein in high bulk tank SCC milk enhances the extension of the rennet coagulation time (RCT) and curd firming time (k20) because there are more serum proteins and the stability of casein micelles are reduced as a result of hydrolysis. Those changes in turn led to poor syneresis, lower cheese yield, increased moisture content and lower fat and protein content in cheese. Finally, there is a favourable relationship between lactose and milk technological properties because the decrease of this component in case of mastitis is associated with an increase of milk pH. Thus, the three parameters, SCC, pH, and lactose affect, contemporarily and independently, milk quality and coagulation properties, and this is why have been highlighted as potential indicators traits for improving cheese-making ability of sheep milk.

Rates of myogenesis and myofiber numbers are reduced in late gestation IUGR fetal sheep.

Intrauterine growth-restricted (IUGR) fetuses are born with reduced skeletal muscle mass. We hypothesized that reduced rates of myogenesis would contribute to fewer and smaller myofibers in IUGR fetal hindlimb muscle compared to the normally growing fetus. We tested this hypothesis in IUGR fetal sheep with progressive placental insufficiency produced by exposing pregnant ewes to elevated ambient temperatures from 38 to 116 days gestation (dGA; term = 147 dGA). Surgically catheterized control (CON, n = 8) and IUGR (n = 13) fetal sheep were injected with intravenous 5-bromo-2′-deoxyuridine (BrdU) prior to muscle collection (134 dGA). Rates of myogenesis, defined as the combined processes of myoblast proliferation, differentiation, and fusion into myofibers, were determined in biceps femoris (BF), tibialis anterior (TA), and flexor digitorum superficialis (FDS) muscles. Total myofiber number was determined for the entire cross-section of the FDS muscle. In IUGR fetuses, the number of BrdU+ myonuclei per myofiber cross-section was lower in BF, TA, and FDS (P < 0.05), total myonuclear number per myofiber cross-section was lower in BF and FDS (P < 0.05), and total myofiber number was lower in FDS (P < 0.005) compared to CON. mRNA expression levels of cyclins, cyclin-dependent protein kinases, and myogenic regulatory factors were lower (P < 0.05), and inhibitors of the cell cycle were higher (P < 0.05) in IUGR BF compared to CON. Markers of apoptosis were not different in IUGR BF muscle. These results show that in IUGR fetuses, reduced rates of myogenesis produce fewer numbers of myonuclei, which may limit hypertrophic myofiber growth. Fewer myofibers of smaller size contribute to smaller muscle mass in the IUGR fetus.

Increased susceptibility to Haemonchus contortus infection by interleukin-5 modulation of eosinophil responses in sheep.

Eosinophils are prominent effector cells in immune responses against gastrointestinal nematode infections in ruminants, but their in vivo role has been hard to establish in large animals. Interleukin-5 is a key cytokine in the induction and stimulation of anti-parasitic eosinophil responses. This study attempted to modulate the eosinophil response in sheep through vaccination with recombinant interleukin-5 (rIL-5) and determine the effect on subsequent Haemonchus contortus infection. Nematode-resistant Canaria Hair Breed (CHB) sheep vaccinated with rIL-5 in Quil-A adjuvant, had lower blood eosinophil counts and higher mean worm burdens than control sheep vaccinated with Quil-A adjuvant alone. In addition, adult worms in IL-5-vaccinated sheep were significantly longer with higher eggs in utero in female worms, supporting an active role of eosinophils against adult parasites in CHB sheep. These results confirm that eosinophils can play a direct role in effective control of Hcontortus infection in sheep and offer a new approach to study immune responses in ruminants.

The efficacy of pirfenidone in a sheep model of pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive fibrotic lung disease with unknown cause. While the drugs nintedanib and pirfenidone have been approved for the treatment of IPF, they only slow disease progression and can induce several side-effects, suggesting that there is still an unmet need to develop new efficacious drugs, and interventions strategies, to combat this disease. We have recently developed a sheep model of pulmonary fibrosis for the preclinical testing of novel anti-fibrotic drugs. The aim of this study was to assess the effects of pirfenidone to ascertain its suitability as a benchmark for comparing other novel therapeutics in this sheep model. To initiate localized fibrosis, sheep were given two infusions of bleomycin (0.6 U/ml per infusion), a fortnight apart, to a specific lung segment. The contralateral lung segment in each sheep was infused with saline to act as an internal control. Two weeks after the final bleomycin infusion, either pirfenidone or methylcellulose (vehicle control) were administered orally to sheep twice daily for 5 weeks. Results showed that sheep treated with pirfenidone had improved lung function, ameliorated fibrotic pathology, lower numbers of active myofibroblasts, and reduced extra cellular matrix deposition when compared with the relevant measurements obtained from control sheep treated with vehicle. This study showed that pirfenidone can attenuate bleomycin-induced pulmonary fibrosis in sheep, and can therefore be used as a positive control to assess other novel therapeutics for IPF in this model.

Impaired Baroreflex Function in an Ovine Model of Chronic Heart Failure Induced by Multiple Coronary Microembolizations.

Testing new therapies in heart failure (HF) requires a chronic stable model of HF in large animals. Microembolization of the coronary arteries has been used to model HF previously; however, neural control has not been previously explored in this model. Thus the aim of this study was to further characterize neural control in this model of HF. HF was induced by infusion of microspheres (45 micron; 1.3 ml) into the proximal left coronary artery or left descending coronary arteries, with three sequential embolizations over 3 weeks. Twelve to 14 weeks after the final embolization, and when ejection fraction had decreased below 45%, animals were instrumented to record blood pressure and heart rate. Baroreflex control of heart rate was investigated in conscious animals. Additionally, pressure-volume loops were constructed under anesthesia. Embolization-induced HF was associated with a decrease in mean arterial pressure (67 ± 2 vs. 85 ± 4 mmHg, p < 0.05), an increase in heart rate (108 ± 4 vs. 94 ± 4 bpm, p < 0.05), and a significant increase in left ventricular end-diastolic pressure (11.4 ± 2 vs. 6.2 ± 1 mmHg, p < 0.01). Under conscious conditions, there was a significant decrease in the gain (−8.2 ± 2 vs. −4.1 ± 1 beats/min/mmHg, p < 0.05) as well as the lower plateau of the baroreflex in HF compared to control animals. HF was also associated with significantly increased respiratory rate (107 ± 4 vs. 87 ± 4 breaths/min, p < 0.01) and incidence of apneas (520 ± 24 vs. 191 ± 8 apnea periods >4 s, p < 0.05), compared to control sheep. The microembolization model of heart failure is associated with an increase in left ventricular end-diastolic pressure, impaired cardiac function, and altered baroreflex control of the heart. These findings suggest this chronic model of HF is appropriate to use for investigating interventions aimed at improving neural control in HF.

Detection of Echinococcus granulosus sensu lato infection by using extracts derived from a protoscoleces G1 cell line.

Cystic echinococcosis (CE) can be diagnosed by means of several serological approaches, but their results vary among laboratories due to the molecular characteristics of the reference antigens used. Thus, this study aimed to address both the relevance of an EGPE cell line previously obtained from Echinococcus granulosus protoscoleces G1 and the complexity of the immune response by using two different in vitro growth stages as separate sources of parasite antigens. The serum reactivity was investigated by western blotting (WB) in 21 CE patients from an endemic area in a matched case-control design and also in seven experimentally infected sheep and five healthy control sheep. EGPE-antigen-human serum sensitivity by WB was higher than that of hydatid fluid (HF) WB, ELISA and DD5 (P<.05, Chi-square test). EGPE protein extract was immunogenic in mice and hyperimmune plasma reacted with HF proteins, and AgB2 expression was detected by molecular analysis. Proteins of 37 to 60kDa were recognized by 95.24% of the CE patients' sera but, with poor specificity. Statistically significant differences were found between serum protein extract recognition at 7 and 20days of cell growth. The EGPE cell line is a laboratory source of antigens for improvement of CE serological diagnosis.

Effects of Mannan Oligosaccharides on Gas Emission, Protein and Energy Utilization, and Fasting Metabolism in Sheep.

Simple SummaryMannan oligosaccharides (MOS) are a promising feed additive to improve animal health, immune capacity, and antioxidation. Based on the previous studies, we carried out three experiments to investigate the effects of MOS on the gas emission, protein and energy utilization, and fasting metabolism of sheep. The results showed that 2.0% MOS supplementation led to the lowest in vitro CO2 production and lower CH4 production and decreased in vivo intake. However, it also decreased urine nitrogen excretion and energy released as CH4, and then improved the utilization of crude protein and energy of sheep. There were no differences in the parameters of respiration and energy metabolism of sheep under the fasting condition. The findings indicated that MOS slightly affected the gas emission and nutrients and energy utilization of sheep.This study investigated the effects of mannan oligosaccharides (MOS) on in vitro and in vivo gas emission, utilization of crude protein (CP) and energy, and relative parameters of sheep under fasting metabolism conditions. In vitro gas productions were evaluated over 12 h in sheep diets containing different amounts of MOS (from 0% to 6.0%/kg, the increment was 0.5%). A control experiment was used to assess the gas emission, utilization of CP and energy, and fasting metabolism in control sheep and sheep treated with 2.0% MOS over 24 days (d). The results showed that 2.0% MOS supplementation led to the lowest in vitro CO2 production and less CH4 production, while also leading to decrease in vivo nutrients intake, CP and energy excretion, digested and retained CP, and energy released as CH4 (p < 0.05). Furthermore, 2.0% MOS supplementation appeared to decrease in vivo O2 consumption and CH4 production per metabolic body weight (BW0.75), and increase the CP retention rate of sheep (p < 0.074). MOS did not affect other parameters, along with the same parameters of sheep under fasting metabolism conditions (p > 0.05). The findings indicate MOS has only slight effects on the gas emission and nutrients and energy metabolism of sheep.

Detection of the Bacteroides fragilis toxin gene in sheep with and without small intestinal adenocarcinoma

ABSTRACTAims: To determine if presence of the Bacteroides fragilis toxin (bft) gene, a molecular marker of colonic carriage of entertoxigenic Bacteroides fragilis (ETBF) in humans, was associated with a finding of small intestinal adenocarcinomas (SIA) in sheep in New Zealand.Methods: Samples of jejunal tissue were collected from the site of tumours and from grossly normal adjacent tissue in 20 sheep, in different consignments, diagnosed with SIA based on gross examination of viscera following slaughter. Two jejunal samples were also collected from a control sheep in the same consignment that had no gross evidence of SIA. A PCR assay was used to detect the presence of the bft gene in the samples.Results: Of the sheep with SIA, the bft gene was amplified from one or both samples from 7/20 (35%) sheep, and in sheep that had no gross evidence of SIA the bft gene was amplified from at least one sample in 11/20 (55%) sheep (RR 0.61; 95% CI = 0.30–1.25; p = 0.34). Of 11 positive samples analysed, ETBF subtype bft-1 was detected in one, bft-2 was detected in 10, and none were bft-3.Conclusions and Clinical Relevance: There was a high prevalence of detection of the bft gene in both SIA-affected and non-affected sheep, but there was no apparent association between carriage of ETBF, evidenced by detection of the bft gene, and the presence of SIA. ETBF are increasingly implicated in the aetiology of human colorectal cancer, raising the possibility that sheep may provide a zoonotic reservoir of this potentially carcinogenic bacterium.Abbreviation: Bft: Bacteroides fragilis toxin; ETBF: Enterotoxigenic Bacteroides fragilis; SIA: Small intestinal adenocarcinoma

Assessment of different pain management methods for the treatment of claw lesions in Meat Merino ewes

Pain management during veterinary procedures is a significant component of animal welfare and has legal as well as ethical implications. Even though regional intravenous anaesthesia (RIVA) is an accepted method for painful procedures involving the distal digits of sheep, this anaesthetic technique is rarely applied in the field. The primary goal was to investigate the feasibility, safety and efficacy of RIVA in sheep. A secondary goal was to examine whether the anaesthetic procedure can be improved by combining RIVA with sedation and whether these methods have a positive effect on the postoperative wellbeing of the animals. A total of 36 Meat Merino sheep with contagious interdigital dermatitis and 12 healthy control sheep were used. Behaviour was observed during treatment of the lame sheep using various pain management protocols and during routine claw trimming of the healthy sheep, and all the sheep were observed after the procedures. The observed behaviours were assessed using scores and the scores compared among the animals of the 4 study groups (RIVA, sedation with xylazine hydrochloride + RIVA, placebo, control). RIVA was successfully conducted in sheep. Local reactions at the application sight and in the tourniquet area in 2 animals resolved completely. A significant reduction in defensive movements during the painful procedure confirmed the efficacy of RIVA. Stress-associated behaviours, including head shaking and idle chewing, occurred with a similar frequency in RIVA and placebo animals, leading to the conclusion that stress levels due to the handling in dorsal recumbency were comparable between these 2 groups. Sedation reduced the frequency of pain- and stress-associated behaviours, including guarding, favouring limbs, vocalisation, idle chewing and bruxism. Xylazine hydrochloride-RIVA animals displayed better weight-bearing in the affected limb, better food uptake and ruminated more postoperatively than sheep from the other study groups. RIVA in sheep is straightforward, safe and effective. Additional sedation reduces the stress and pain response. This pain and stress management has a positive effect on the postoperative wellbeing of sheep.

Developmental Programming: Contribution of Epigenetic Enzymes to Antral Follicular Defects in the Sheep Model of PCOS

Prenatal testosterone (T)-treated sheep, similar to women with polycystic ovary syndrome (PCOS), manifest oligo-/anovulation, hyperandrogenism, and polyfollicular ovary. The polyfollicular ovarian morphology, a result of persistence of antral follicles, arises, in part, by transcriptional changes in key mediators of follicular development that, in turn, are driven by epigenetic mechanisms. We hypothesized that prenatal T excess induces, in a cell-specific manner, transcriptional changes in key mediators of follicular development associated with relevant changes in epigenetic machinery. Expression levels of key mediators of follicular development, DNA methyltransferases (DNMTs), and histone de-/methylases and de-/acetylases were determined in laser-capture microdissection-isolated antral follicular granulosa and theca and ovarian stromal cells from 21 months of age control and prenatal T-treated sheep (100 mg IM twice weekly from gestational day 30 to 90; term: 147 days). Changes in histone methylation were determined by immunofluorescence. Prenatal T treatment induced the following: (i) cell-specific changes in gene expression of key mediators of follicular development and steroidogenesis; (ii) granulosa, theca, and stromal cell-specific changes in DNMTs and histone de-/methylases and deacetylases, and (iii) increases in histone 3 trimethylation at lysine 9 in granulosa and histone 3 dimethylation at lysine 4 in theca cells. The pattern of histone methylation was consistent with the expression profile of histone de-/methylases in the respective cells. These findings suggest that changes in expression of key genes involved in the development of the polyfollicular phenotype in prenatal T-treated sheep are mediated, at least in part, by cell-specific changes in epigenetic-modifying enzymes.

Impacts of breed type and vaccination on Teladorsagia circumcincta infection in native sheep in Gran Canaria

Vaccines and genetic resistance offer potential future alternatives to the exclusive use of anthelmintics to control gastrointestinal nematodes (GIN). Here, a Teladorsagia circumcincta prototype vaccine was administered to two sheep breeds which differ in their relative levels of resistance to infection with GIN. Vaccination of the more susceptible Canaria Sheep (CS) breed induced significant reductions in worm length and numbers of worm eggs in utero (EIU) when compared to control CS sheep. In the more resistant Canaria Hair Breed (CHB), although vaccination induced a reduction in all parasitological parameters analysed, differences between vaccinated and control sheep were not statistically significant. Such interactions between sheep breed and vaccination may allow better integrated control of GIN in future.

Use of grape residue flour in lactating dairy sheep in heat stress: Effects on health, milk production and quality.

The objective of this study was to evaluate the effects of grape residue flour (GRF) on antioxidant activities, biochemistry variables, components of the immune system and milk production and quality of Lacaune sheep in heat stress. Twenty-seven multiparous lactating sheep [50 ± 1.8 days (d) milking] were stratified by initial body weight, age, date of lambing and milk production and assigned randomly to 1 of 3 treatments (9 sheep/treatment): no GRF supplementation (control group) or supplementation at 1% (10 g/kg GRF) or 2% (20 g/kg GRF) of GRF (bark and seed) in the concentrate (grains and minerals mixture). Each ewe received 0.8 kg/d of concentrate, 3.6 kg/d of corn silage, and 0.25 kg/d of Cynodon spp hay. Milk production along with blood and milk samples were collected on d 1, 10 and 15. The 2% GRF sheep had increased serum concentrations of superoxide dismutase and glutathione peroxidase activity on d 15 compared to control sheep. Over time (d 10 to 15), lipid peroxidation was reduced in 2% GRF sheep. Total serum antioxidant capacity was greater in 2% GRF sheep compared to control sheep on d 10 and 15. Superoxide dismutase and glutathione peroxidase activity in milk samples were greater in 2% GRF sheep compared to control sheep. Supplementation with GRF did not affect milk production but GRF sheep were more efficient compared to control sheep. Protein and lactose concentrations were similar between treatments, but total solids and fat concentrations were greater in 2% GRF sheep compared to control sheep on d 15. Somatic cell count was reduced in GRF sheep compared to control sheep. In summary, supplementation with 2% GRF in dairy sheep in heat stress resulted in antioxidant and anti-inflammatory responses, which improved milk quality and reduced somatic cell count and lipid peroxidation.

Evaluation of in vitro neutralization of epoxyscillirosidine by antibodies raised in sheep

Intoxication by Moraea pallida Bak. (yellow tulp) in livestock is of great importance in South Africa, ranking top among all plant-induced cardiac glycoside toxicosis. The toxic principle, a bufadienolide, is 1α, 2α-epoxyscillirosidine. Treatment of poisoning is challenging and affected livestock often succumbs due to the stress of handling. Manipulating animals to resist poisoning is a potential management strategy. The goal of this study was to explore the potential to develop a vaccine against epoxyscillirosidine by raising antibodies against epoxyscillirosidine in sheep and to assess the neutralization ability of the antibodies in vitro. Epoxyscillirosidine was successfully conjugated to keyhole limpet haemocyanin (KLH) and bovine serum albumin (BSA) rendering them immunogenic. The sheep, vaccinated with epoxyscillirosidine-KLH conjugate (n = 4) and KLH (n = 2) with Montanide, developed antibodies as determined with an indirect enzyme linked immunosorbent assay (ELISA). Total immunoglobulins from sera of vaccinated and control sheep that were purified and concentrated using ammonium sulphate precipitation were 11,940 and 7850 μg, respectively. The in vitro neutralization assay using the methyl blue tetrazolium bromide (MTT) cell viability assay indicated no significant difference (p > 0.05) between anti-epoxyscillirosidine-KLH and KLH antibodies. Rather, the antibodies seemed to enhance the cytotoxicity of epoxyscillirosidine in H9c2 cells. Thus, it is necessary to develop improved vaccination methods to generate antibodies capable of neutralizing the functional group responsible for epoxyscillirosidine toxicity.

Microscopic alterations in Fasciola hepatica from sheep treated with albendazole.

Currently, albendazole is one of the most commonly used drugs because of its affordability. The objective was to evaluate the histopathology of Fasciola hepatica specimens. For this, the efficacy test was performed on sheep treated with albendazole at the dose recommended for F. hepatica, in which the helminths recovered at necropsy were counted and separated for histology. Spermatogenic cells from parasites recovered from treated and control sheep were examined by microscopy. The fecal egg-count reduction test revealed 97.06% efficacy of albendazole in the treatment of F. hepatica. Changes in testicular tubule cells started 48 hours after treatment and became evident within 72 hours, at which point it became difficult to identify cell types. Primary and secondary spermatogonia became increasingly rare and intercellular vacuolization was more evident. Signs of apoptosis, with pycnotic nuclei and evidence of keriorrexia were observed at all times. Cell debris was identified 96 hours after treatment. The results indicated that parasitic spermatogenesis was severely affected by albendazole and demonstrated the importance of the use of histopathology for the diagnosis of therapeutic efficacy in field strains.