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PubMed2004

Discrimination between scrapie and bovine spongiform encephalopathy in sheep by molecular size, immunoreactivity, and glycoprofile of prion protein.

A procedure for discrimination between scrapie and bovine spongiform encephalopathy (BSE) in sheep is of importance for establishing whether BSE has entered the sheep population. Since BSE has not yet been found in sheep at the farm level, such discrimination procedures can be developed only with experimental sheep BSE. Two distinctive molecular features of the prion protein (PrP)-molecular size and glycosylation profile-in proteinase K digests of brain stem tissue from sheep were used here; upon Western blotting, these features led to an unequivocal discrimination among natural scrapie, experimental scrapie, and experimental BSE. The higher electrophoretic mobility of PrP in sheep BSE could be best observed after deglycosylation treatment with N-glycosidase F. A simpler method for confirmation of this size difference involved comparison of the ratios for the binding of two monoclonal antibodies: P4 and 66.94b4. Based on epitope mapping studies with P4 and peptides, it appeared that N-terminal amino acid sequence WGQGGSH was intact only in sheep scrapie digests. Another feature typical for PrP in sheep BSE was the large fraction of diglycosylated PrP (70% or more). These data were obtained for a large group of positive sheep, consisting of 7 sheep with experimental BSE infection (genotypes: six ARQ/ARQ and one AHQ/AHQ), 48 sheep naturally infected with scrapie (six different genotypes), and 3 sheep with primary experimental scrapie infection. Routine tests of slaughter material serve well for the initial detection of both BSE and scrapie. With Western blotting as a rapid follow-up test, a 66.94b4/P4 antibody binding ratio above 1.5 is a practical indicator for serious suspicion of BSE infection in sheep.

Amino Acid Sequence↗
PubMed1987

Glucose-dose dependent characteristics of insulin secretion in obese and lean sheep.

We previously reported that obesity in sheep and cattle was associated with basal hyperinsulinemia, insulin resistance, and an exaggerated insulin response to a single dose (350 mg/kg) of glucose. In this study, the glucose-dose dependency of insulin secretion in obese and lean sheep was determined by 1) using jugular venous concentrations of insulin (Exp 1) and 2) arteriovenous differences in insulin concentrations across the pancreas together with plasma flow rates in the portal vein (Exp 2). Sheep were injected with glucose doses of 0 (water), 10, 30, 100, and 350 mg glucose/kg body weight in Exp 1 (six sheep per group) and with a low (20 mg/kg) and high (200 mg/kg) dose of glucose in exp 2 (four sheep per group). In Exp 1, mean (+/- SE) pretreatment plasma concentrations of insulin (22.0 +/- 1.7 vs. 9.4 +/- 0.4 microU/ml) and glucose (56.1 +/- 0.5 vs. 52.4 +/- 0.8 mg/dl) were greater (P less than 0.01) in obese than lean sheep fasted for 12 h. The glucose-induced rises in insulin concentrations above pretreatment levels were always greater (P less than 0.05) in obese than lean sheep regardless of glucose dose. Eadie-Scatchard plot analysis of the hyperbolic relationship between the acute insulin and acute glucose response areas (0 to +10 min) indicated that the maximum (Vmax) early phase insulin response was greater (P less than 0.025) in obese than lean sheep (568 +/- 148 vs. 156 +/- 33 microU ml-1 X min). In Exp 2, pretreatment concentrations of insulin (25.1 +/- 3.4 vs. 5.6 +/- 1.2 microU/ml) and glucose (58.3 +/- 1.8 vs. 45.5 +/- 1.1 mg/dl) in arterial plasma were greater (P less than 0.01) in obese than in lean sheep fasted 18 to 22 h. Similarly, pretreatment pancreatic secretion rates of insulin were greater (P less than 0.01) in obese (17.8 +/- 5.8 mU/min) than in lean (4.9 +/- 1.3 mU/min) sheep. Glucose-induced acute (0 to +10 min) increments in pancreatic secretory rates of insulin also were greater (P less than 0.05) in obese than in lean sheep after the low (215 +/- 73 vs. 11 +/- 15 mU) and high (881 +/- 281 vs. 232 +/- 66 mU) doses of glucose. It was concluded that insulin secretion in response to a range of stimulatory concentrations of glucose was greater in obese than in lean sheep because the obese sheep had greater maximum (i.e. Vmax) acute phases of glucose-induced insulin secretion.

Animals↗
PubMed1997

[Serological investigations in the diagnosis of viral infections derived from cattle in sheep].

In the coast region of the north-west of Germany 269 arbitrary chosen blood samples of 22 flocks of sheep were tested for the existence of BHV-1, BVD and PI-3 antibodies. The livestock of 13 farms with 163 sheep consistent of sheep and bovine, 9 herds with 106 sheep kept sheep only. The results show that BHV-1 antibodies were not present in the sampled sheep. Contrary to this are the results for BVD and PI-3. The blood tests for BVD antibodies were positive in 6.7% of the samples. This result indicates a low prevalence rate of BVD virus in the investigated region. The positive results showed distinct differences in distribution for the two farm types. Out of all BVD antibody positive reactors (n = 18) 83% were located in 7 farms keeping sheep and cattle. The rest of the positive reactors (17%) was found on 2 farms keeping sheep only. For PI-3 the distribution of farms keeping sheep plus cattle vrs. keeping sheep only concerning the antibody positive stocks was 7 positive herds out of 13 vrs. 6 positive herds out of 9. Again within the sheep herds of the two farms types occurred significant differences. Farms with sheep and cattle had 19% to 76% PI-3 antibody positive blood samples and farms with sheep only 1% to 11%.

Animals↗
PubMed1994

Age-dependent immune response in Merino sheep.

The numerical and functional attributes of populations of lymphocytes were compared in the blood, lymph and skin of young and mature sheep. Young sheep, four to eight months old, had a lower proportion of CD4+ cells in blood, lymph and skin than mature sheep three to six years old. In contrast, B cells and T19+ cells were as prevalent or more prevalent in young sheep as in mature sheep. Blood lymphocytes from young sheep, cultured in vitro produced less interferon-gamma, both spontaneously and in the presence of concanavalin A than did lymphocytes from older sheep. The serum antibody responses of adult sheep to the T cell-independent antigen Brucella abortus lipopolysaccharide (LPS) were greater over a range of antigen doses, suggesting that an apparent excess of antigen could not overcome the relative immune deficiency of young sheep. The adjuvant Quil A corrected the depressed antibody response of young sheep to B abortus LPS, but dextran sulphate did not. The skin contact hypersensitivity of mature sheep to dinitrochlorobenzene was greater. However, the T cell phenotypes present in infiltrates of lymphocytes elicited by the intradermal injection of tetanus and diphtheria, but not tuberculin antigens, were comparable for the two age groups. The capacity of Quil A to raise the antibody responses of both young and mature sheep to a similar titre suggests that it may be possible to overcome the immunological hyporesponsiveness that may contribute to the disease susceptibility of young sheep.

Aging↗
PubMed1983

Functional depression of H2 histamine receptors in sheep with experimental allergic asthma.

We tested the hypothesis that airway hyperresponsiveness to histamine in the allergic sheep is related to functional depression of H2 histamine receptors. Thirteen of 32 sheep responded with bronchoconstriction to inhaled Ascaris suum antigen (allergic sheep), and the remainder served as controls (nonallergic sheep). In the allergic sheep, 50 and 100 breaths of 5% histamine solution increased mean pulmonary resistance (RL) to 235% and 438% of baselines, respectively. The corresponding values in nonallergic sheep were 200% and 211%, indicating a greater response to the higher dose of histamine in allergic sheep. Selective H1-receptor stimulation with 50 breaths of histamine (pretreatment with the H2-receptor antagonist metiamide) failed to enhance the effect of histamine in allergic sheep (mean RL increased to 239% of baseline) whereas it enhanced the histamine response in nonallergic sheep (RL increased to 438% of baseline). Selective H2-receptor stimulation (pretreatment with the H1-receptor antagonist chlorpheniramine) caused histamine to decrease RL by 31% in the nonallergic sheep group; it blocked but did not reverse the histamine effect in the allergic sheep. Similar observations were made in a different group of animals when selective H1- or selective H2-receptor stimulation was produced by 100 breaths of histamine. The cutaneous wheal response to intradermal histamine dilutions of 0.0001. 0.001, 0.01, and 1 mg/ml was similar in both groups. In nonallergic sheep, both chlorpheniramine and metiamide blunted the cutaneous wheal response. In allergic animals, only chlorpheniramine blunted the cutaneous wheal response, whereas metiamide was without effect. We conclude that airway hyperresponsiveness to histamine in allergic sheep is related to a functional depression of H2 receptors and that such a defect is observed both in the airways as well as in the skin.

Animals↗
PubMed1986

Comparative pathogenicity and antigenic cross-reactivity of Rift Valley fever and other African phleboviruses in sheep.

Homologous and heterologous haemagglutination-inhibition (HAI), complement-fixation (CF), immunodiffusion (ID) and mouse neutralization tests were performed with the Lunyo (LUN) and a Zimbabwean strain of Rift Valley fever (RVF) virus, the prototype and a South African strain of Arumowot (AMT) virus and prototype strains of Gordil (GOR), Saint-Floris (SAF) and Gabek Forest (GF) viruses, using immune mouse ascitic fluids prepared against these viruses. Reactions of identity occurred in all tests between LUN and the Zimbabwean strains of RVF and between the two strains of AMT virus. Otherwise, cross-reactions occurred between all the phleboviruses in HAI tests, while reactions in CF, ID and neutralization tests were monospecific for virus serotypes, except that weak cross-reaction occurred between GOR and SAF viruses in CF and ID tests. Four sheep infected subcutaneously with the Zimbabwean strain of RVF virus developed transient fever, viraemia, leucopaenia, relative thrombocytopaenia, haemoconcentration and raised serum enzyme levels, which indicated that the sheep had developed necrotic hepatitis. Disseminated focal necrotic hepatitis was confirmed in a sheep killed for examination on day 4 post-infection. The other three sheep recovered uneventfully after only mild depression and anorexia. Groups of three sheep infected with SAF, GOR, AMT and GF viruses had no demonstrable viraemia or other sign of infection or illness, except that the sheep infected with AMT developed mild fever lasting less than 24 h. Antibody responses were monitored at intervals over a period of 24 weeks in all sheep by homologous and heterologous HAI, CF and cell culture neutralization (CPENT) tests. Homologous antibody responses were marked in the RVF-infected sheep and their sera cross-reacted strongly in HAI tests with antigens of the other viruses. The sera of the RVF-infected sheep cross-reacted less markedly in CF and CPENT tests. Homologous antibody responses were poor in all the sheep infected with phleboviruses other than RVF, and the cross-reactivity of their sera for RVF antigen or virus was negligible. All sheep were challenged with RVF virus 48 weeks after their initial infection. The sheep which had originally been infected with RVF virus were immune and developed neither fever nor viraemia. All other sheep developed fever, viraemia and antibodies to RVF virus. It was concluded that the African phleboviruses, other than RVF, are unlikely to cause disease in livestock or to induce antibodies which could cause confusion in the diagnosis of RVF.

Animals↗
PubMed1998

New gene assignments using a complete, characterized sheep-hamster somatic cell hybrid panel.

The generation and characterization of new sheep-hamster cell hybrids is reported from the fusion of sheep white blood cells with six different hamster auxotrophs. Selection from these and previously generated cell hybrids has led to the production of a panel of 30 hybrids covering the complete sheep genome of 28 chromosomes. Over half of the cell hybrids in this panel contain single sheep chromosomes. By complementation, the following new assignments have been made using the panel: phosphoribosyl N-formylglycinamide amidotransferase (PRFGA) to sheep chromosome (chr) 11; adenylosuccinate synthetase (ADSS) to sheep chr 12; adenylosuccinate lyase (ADSL) to sheep chr 3q; 3-hydroxy-3-methylglutaryl-coenzyme A synthase (HMGCS) to sheep chr 16; dihydrofolate reductase (DHFR) to sheep chr 5; and adenine phosphoribosyltransferase (APRT) to sheep chr 14. The gene phosphoribosylaminoinidazole-carboxamide formyltransferase/Inosinicase (PRACFT) has now been regionally assigned to chr 2q. By isozyme analysis, phosphogluconate dehydrogenase (PGD) was assigned to sheep chr 12, anchoring the sheep syntenic group U1 to this chromosome, and mannose phosphate isomerase (MPI) was assigned to sheep chr 18. Furthermore, the chromosomal assignment of 110 microsatellites was confirmed using this cell panel.

Adenine Phosphoribosyltransferase↗
PubMed1993

Exaggerated cardiopulmonary response after bacteremia in sheep with week-old thermal injury.

OBJECTIVE: To determine if thermal injury impairs pulmonary intravascular clearance of bacteria and therefore leads to exaggerated cardiopulmonary dysfunction in sheep, since endotoxin infusion has been previously shown to induce more severe pulmonary injury after thermal injury. DESIGN: Prospective, unblinded, randomized, controlled trial. SETTING: Laboratory at a large university medical center. INTERVENTIONS: Chronically instrumented, anesthetized sheep received a 40% total body surface area, third-degree thermal injury. Live Pseudomonas aeruginosa (10(7) P. aeruginosa/min for 1 hr; n = 6) were infused 7 to 10 days after thermal injury. Similarly prepared noninjured sheep received the same pseudomonas infusion (n = 7) or saline (n = 7). MEASUREMENTS AND MAIN RESULTS: Bacterial clearance, which measures phagocytosis by the pulmonary intravascular macrophages, was equally efficient in intact sheep and sheep with thermal injury. Pulmonary hypertension persisted for 18 hrs after thermal injury, compared with 8 hrs in noninjured sheep. Lung lymph flow significantly increased from 6 to 8 hrs in only the thermal injury group. Both bacteremic groups developed a hyperdynamic circulation from 6 to 8 hrs, but cardiac index was 1 to 1.5 L/min/m2 higher in thermally injured sheep. Total peripheral resistance index decreased significantly from 6 to 24 hrs in thermally injured sheep and from 6 to 12 hrs in intact bacteremic sheep. Mean arterial pressure of thermally injured sheep was increased at baseline and for the first 6 hrs compared with noninjured animals. Mean arterial pressure decreased from 6 to 24 hrs in sheep with thermal injury but did not change in intact bacteremic sheep. CONCLUSIONS: Bacterial clearance was not impaired by preceding thermal injury in sheep. Bacteremia in the presence of a preexisting thermal injury led to more persistent pulmonary hypertension and an exaggerated hyperdynamic circulation.

Animals↗
PubMed2001

Comparison between sheep and human cervical spines: an anatomic, radiographic, bone mineral density, and biomechanical study.

STUDY DESIGN: The quantitative anatomic, radiographic, computerized tomographic, and biomechanical data of sheep and human cervical spines were evaluated. OBJECTIVES: To compare the anatomic, radiographic, computerized tomographic, and biomechanical data of human and sheep cervical spines to determine whether the sheep spine is a suitable model for human spine research. SUMMARY OF BACKGROUND DATA: Sheep spines have been used in several in vivo and in vitro experiments. Quantitative data of the normal sheep cervical spine are lacking, yet these data are crucial to discussion about the results of such animal studies. METHODS: In this study, 20 fresh adult female Merino sheep cervical spines and 20 fresh human cadaver cervical spines were evaluated anatomically, radiographically, computerized tomographically, and biomechanically. Three linear and two angular parameters were evaluated on four digital radiographic views: anteroposterior, right lateral in neutral position, flexion, and extension. Quantitative computed tomography scans at the center of each vertebral body and 3 mm below both endplates were analyzed for bone mineral density measurements. Biomechanical testing was performed in flexion, extension, axial rotation, and lateral bending by a nondestructive stiffness method using a nonconstrained testing apparatus. Range of motion and stiffness of each motion segment were calculated. Additionally, 10 linear anatomic parameters of each vertebra were measured using a digital ruler. RESULTS: Anterior and mean disc space height in the sheep cervical spine increased constantly from C2-C3 to C6-C7, whereas middle disc space height decreased and posterior disc space height remained unchanged. Anterior and mean disc space height were significantly higher in sheep. In both sheep and human cervical spines, intervertebral angles were not significantly different. Standard deviations of bone mineral density in the human cervical spine were fourfold higher than in the sheep cervical spine, yet no significant differences were found in bone mineral density values between the two species. Range of motion differed significantly between the two species except in flexion-extension of C3-C4, C5-C6, axial rotation of C2-C3, and lateral bending of C2-C3, C3-C4, and C4-C5. Stiffness also was significantly different except in flexion-extension of C2-C3, C4-C5, C5-C6, and lateral bending of C2-C3, C3-C4, and C4-C5. Anatomic evaluation showed no difference in upper endplate parameters for C4 and C5. CONCLUSIONS: Although several differences were found between human and sheep cervical spines, the small intergroup standard deviations and the good comparability with the human spine encourage the use of the sheep cervical spine as a model for cervical spine research. On the basis of the quantitative data obtained in this study, the sheep motion segment C3-C4 seemed to be the most reliable model for the corresponding human motion segment.

Adult↗
PubMed2004

Prion protein gene polymorphisms in healthy and scrapie-affected sheep in Greece.

A total of 216 local crossbred sheep from 16 scrapie-affected Greek flocks and 210 purebred sheep of the milk breeds Chios and Karagouniko from healthy flocks were analysed for scrapie-linked polymorphisms in the prion protein (PrP) gene. Of the 216 sheep in this case-control study, 96 sheep were clinical cases, 25 subclinical cases (asymptomatic at the moment of euthanasia but positive by histopathology and/or ELISA detecting proteinase-resistant PrP) and 95 healthy controls (negative by all evaluations). Polymorphisms at codons 136, 154 and 171 were determined by denaturing gradient gel electrophoresis, followed by RFLP and sequencing. Scrapie, both clinical and subclinical, was associated with the genotypes ARQ/ARQ (88 of 110 sheep of that genotype), ARQ/TRQ (9 of 13), ARQ/AHQ (15 of 38) and VRQ/VRQ (9 of 17). Histopathological lesions were more severe in the clinical cases. Genotypes ARQ/ARR (26 sheep), ARQ/ARK (seven sheep), AHQ/ARR (one sheep), ARH/ARH (one sheep) and ARR/ARH (three sheep) were detected exclusively in healthy control sheep. In the purebred survey, four genotypes were present in the Chios sheep (ARQ/ARQ, ARQ/TRQ, ARQ/AHQ and ARQ/ARR) and four in the Karagouniko sheep (ARQ/ARQ, ARQ/AHQ, ARQ/ARR and ARQ/ARH).

Animals↗
PubMed1998

The regulation of lung liquid absorption by endogenous cAMP in postnatal sheep lungs perfused in situ.

1. The lungs of two groups of lambs aged 0-2 weeks and 6-12 weeks were artificially perfused in situ with warmed and oxygenated sheep blood. The airspaces of the lungs were filled with liquid containing an impermeant tracer to allow estimation of net liquid movement across the pulmonary epithelium at rest and after administration of certain drugs. 2. Dibutyryl cAMP (dB-cAMP, 10-4 M) stimulated the rate of lung liquid (LL) absorption in the lungs of four neonatal sheep aged 9-12 days, from -1.43 +/- 0.2 to -2.75 +/- 0.3 ml h-1 (kg body wt)-1 (P < 0.05, comparison of regression lines by Student's t test), but had no effect in four juvenile sheep aged 6-12 weeks (P > 0.10). 3. Theophylline, a non-selective phosphodiesterase (PDE) inhibitor (5 x 10-4 M), increased LL absorption from a resting rate of -1.55 +/- 0.3 to -3.62 +/- 0.5 ml h-1 kg-1 in the lungs of four sheep aged 1-12 days and from -1.47 +/- 0.3 to -3.73 +/- 0.4 ml h-1 kg-1 in four sheep aged 6-12 weeks (P < 0.05, Student's paired t test). 4. The beta-adrenergic antagonist sotalol (10-4 M) reduced LL absorption rate from -1.47 +/- 0.1 to -1.22 +/- 0.1 ml h-1 kg-1 (P < 0.01) in the lungs of four sheep aged 4-13 days, while theophylline given after sotalol had no effect. In four sheep aged 6-12 weeks, sotalol had no effect on LL absorption rate, whereas theophylline given after sotalol increased LL absorption rate from -1.06 +/- 0.1 to -1.92 +/- 0.2 ml h-1 kg-1 (P < 0.05). 5. The A1/A2 purinergic receptor blocker 7-(beta-chloroethyl) theophylline (CET; given at 5 x 10-6 M and 10-4 M) had no effect on LL absorption rate in the lungs of four sheep aged 6-12 weeks, confirming that theophylline produced its effect of increasing LL absorption by inhibiting PDE hydrolytic activity. 6. The selective PDE IV (cAMP-specific) PDE inhibitor rolipram was given in the perfused lungs of seven sheep aged 6-12 weeks at doses between 10-8 and 10-4 M, increasing LL absorption rate at concentrations of 10-6 M and above; the half-maximal effective concentration was estimated to be 5.9 x 10-7 M. 7. Rolipram (10-5 M) increased LL absorption rate from -1.99 +/- 0.2 to -3.18 +/- 0.5 ml h-1 kg-1 in the perfused lungs of four sheep aged 6-11 days, and from -1.21 +/- 0.4 to -3.45 +/- 0.3 ml h-1 kg-1 in the perfused lungs of four sheep aged 6-12 weeks (P < 0.05). Sotalol (10-4 M) reduced LL absorption rate from -3.39 +/- 0.8 to -2. 18 +/- 0.4 ml h-1 kg-1 (P < 0.05) in four sheep aged 10-14 days, while rolipram given after sotalol had no effect. In four sheep aged 6-12 weeks, sotalol had no effect on resting LL absorption rate, whereas rolipram given after sotalol increased absorption rate from -1.27 +/- 0.1 to -2.02 +/- 0.6 ml h-1 kg-1 (P < 0.05). 8. We conclude that cAMP mediates a component of LL absorption postnatally, and that while beta-adrenergic stimulation was the sole source of endogenous cAMP in neonates, this was not the case in juveniles, in whom cAMP originated, at least in part, from other sources.

Absorption↗
PubMed1990

Water and sodium movements across the ruminal epithelium in fed and food-deprived sheep.

Fluid and electrolyte movements across the ruminal epithelium of sheep were studied using the temporarily isolated rumen technique. The sheep were all subjected to the following treatments: (1) fed sheep (fed twice daily), after rumen emptying, received rumen contents from a fed sheep; (2) food-deprived sheep (two meals were omitted), after rumen emptying, received rumen contents from a food-deprived sheep; (3) fed sheep, after rumen emptying, received rumen contents from a food-deprived sheep; and (4) food-deprived sheep, after rumen emptying, received rumen contents from a fed sheep. Food deprivation led to an increased Na concentration of the rumen fluid while K and Cl concentrations, as well as osmolality, decreased. Plasma Na and osmolality decreased. During the 40 min after the rumen contents were exchanged no net movements of water occurred. Then the sheep were given an intraruminal load of saline which gave rise to a significant net absorption of fluid from the rumens of those sheep which had received rumen contents from fed sheep. The change in composition of the rumen contents after food deprivation impaired the absorption of Na and water from the rumen. Furthermore food deprivation reduced the Na absorptive function of the ruminal epithelium, but not the water permeability.

Analysis of Variance↗
PubMed1982

The effects of thiosalicylic and hydroxyethylethylenediaminetriacetic acids on the absorption and excretion of 54Mn and 65Zn in the duodenally dosed sheep.

Nine sheep, each prepared with a single cannula in the ascending duodenum and a reentrant cannula in the terminal ileum were fed a purified diet deficient in manganese and zinc for a period of 1 month. The sheep were then divided into three equal groups and duodenally dosed with a mixture containing MnSO4, ZnSO4, 54Mn, 65Zn, and water (control), thiosalicylic acid (TSA), or hydroxyethylethylenediaminetriacetic acid (HEDTA). Blood and ileal digesta were sampled for 24 h and total urine and feces were collected for 14 days after dosing. Plasma uptake of 54Mn 1 h after dosing was more than threefold higher in the HEDTA-dosed sheep than in other sheep, indicating higher absorption of 54Mn. This was accompanied by a lower flow of 54Mn through the terminal ileum during the first 12 h after dosing. However, total 14-day urinary and fecal excretion of 54Mn and 65Zn were apparently higher in the HEDTA-dosed sheep than in other sheep, each approaching 100% of the dose. Excretion of 54Mn and 65Zn in TSA-dosed sheep was apparently higher than in control sheep. Plasma uptake 65Zn was higher in TSA-dosed sheep and lower in HEDTA-dosed sheep than in control sheep. However, total 14-day excretion of 65Zn in feces and urine was slightly higher in TSA-dosed than in control sheep. It was concluded that chelation of Mn and Zn with HEDTA or TSA would aggravate rather than alleviate a Mn or Zn deficiency in sheep.

Animals↗
PubMed1988

Lung injury and neutrophil density during air embolization in sheep after leukocyte depletion with nitrogen mustard.

The hypothesis that neutrophils are a major mediator of acute lung injury is based in part on the results from leukocyte depletion studies in which peripheral leukopenia is induced by administration of chemotherapeutic agents or antileukocyte serum. Because of valid concerns about the specificity and multiplicity of effects these regimens have in vivo, we designed a correlative structure-function study of the impact of one depletion regimen on the intrapulmonary density of neutrophils in two groups of sheep. We gave one group of four awake sheep repeated injections of nitrogen mustard (0.4 mg/kg body weight on the ninth, sixth, and third day before the experiment) to lower circulating leukocyte counts to less than 1% of their normopenic value. The other group of four awake sheep was not treated before the experiment (normopenic sheep). On the day of the experiment, the sheep were anesthetized to measure hemodynamics and lung lymph dynamics, and to take lung tissue for morphologic examination before and during 4 h of continuous venous air embolization. Reduction of circulating leukocytes by greater than 99% of normopenic values only lowered pulmonary intravascular neutrophil density by about 80%. In baseline lung biopsies, neutrophil density in small pulmonary arteries (1 to 0.1 mm in diameter) in the nitrogen-mustard-treated sheep averaged 51 +/- 23 (+/- 1 SD) cells/mm2 of blood area versus 291 +/- 40 in the normopenic sheep. During venous air embolization, neutrophils sequestered in the pulmonary arterial microvessels of both groups (210 +/- 27 cells/mm2 of blood area in the nitrogen-mustard-treated sheep versus 1,217 +/- 49 in the normopenic sheep). Parallel increases in neutrophil density occurred in alveolar capillaries and small pulmonary veins. Neutrophils attached to the intravascular air emboli and to microvascular endothelial cells. Endothelial cell gaps were seen in about 30% (13 gaps in 48 microvessels) of the small arteries in the nitrogen mustard-treated sheep versus about 80% (38 gaps in 48 microvessels) in the normopenic sheep. Lung lymph protein clearance nearly doubled in the nitrogen mustard-treated sheep and tripled in the normopenic sheep during venous air embolization compared with the respective baseline clearance values. We suggest that methods used to deplete circulating leukocytes do not assure removal of neutrophils from the lungs; the remaining neutrophils appear capable of responding to intravascular inflammatory stimuli.

Animals↗
PubMed1990

Failure to maintain interspecific pregnancy after transfer of Dall's sheep embryos to domestic ewes.

Over a 3-year period, 32 Dall's sheep (Ovis dalli dalli) embryos were transferred into 24 domestic sheep (O. aries) recipients and 4 were transferred into 2 Dall's sheep recipients. In the first year, none of the 10 O. aries recipients was diagnosed pregnant. In the following 2 years, 9 (37%) of the domestic sheep recipients were pregnant on Day 18, 8 (33%) on Day 40, 6 (25%) on Day 90 and 4 (16%) on Day 120; 1 aborted at Day 125 and another at Day 145. Pregnancies were established only in ewes that had previously been recipients of Dall's sheep embryos. The 2 remaining pregnant sheep were treated with progesterone from Day 125 until the fetuses were determined to be dead at Day 145. Both of the Dall's sheep recipients (Year 2) established pregnancies; 1 live Dall's sheep lamb was born 174 days after mating. No differences in serum progesterone, oestrone, prostaglandin F-2 alpha metabolites or cortisol concentrations could be detected during pregnancy between recipients carrying Dall's sheep embryos, recipients receiving progesterone treatment or domestic ewes carrying domestic sheep pregnancies. Six fetuses were necropsied (1 at Day 125 and 5 at Day 145-146): all fetuses were premature and had various degrees of hydranencephaly. No significant differences were found when cotyledon numbers were compared among domestic ewes carrying Dall's sheep lambs. Dall's sheep ewes lambing naturally and domestic ewes lambing naturally. These results demonstrate that the transfer of Dall's sheep embryos to domestic ewes results in the establishment but subsequent loss of pregnancy and that these losses occur throughout gestation.

Animals↗
PubMed1975

Changes in the jugular haematocrit of sheep during feeding.

Intact and splenectomized sheep with and without a rumen fistula were used to investigate changes in the jugular blood haematocrit and plasma osmolality during hourly and once-daily feeding regimes. Osmolality was also estimated in the ruminal fluid of fistulated sheep with spleens. Haematocrit decreased in sheep with spleens before they were given a once-daily feed; it increased when these sheep started to feed, reaching a maximum increase of 13% after 30 min of feeding; it decreased during the remaining 45 min of feeding time and usually continued to decrease after feeding stopped. These changes were not due to diurnal influences. Splenectomized sheep fed once daily showed only small decreases in haematocrit before they were fed. Increases occurred with the onset of eating but they were smaller (7%) than in intact sheep and were of shorter duration. In hourly fed sheep with spleens, haematocrit decreased in the early stages of sampling in a manner similar to that for sheep fed once daily. The changes in haematocrit that did occur were not related in any obvious manner to the feeding regime. The haematocrit in splenectomized sheep fed hourly was stable throughout feeding. Variations in the haematocrit in splenectomized sheep, equivalent to a range of 13% in one of them, were observed in a series of blood samples obtained during a 5-h period remote from the feeding time. Large increases occurred in osmolality of ruminal fluid when sheep were fed daily and this was abolished by hourly feeding. Plasma osmolality in sheep fed once daily increased slowly. Maxima occurred after 100 min from the start of eating and were 7% greater than prefeeding values. Only minor changes were observed when these sheep were fed hourly.

Animals↗
PubMed1965

CARBOHYDRATE METABOLISM IN LIVER FROM FOETAL AND NEONATAL SHEEP.

1. During development of the sheep, the activities of UDP-glucose-alpha-glucan glucosyltransferase and UDP-glucose pyrophosphorylase and the glycogen content are highest in the liver of lambs 2 weeks old and considerably lower in liver from adult sheep. 2. The activity of hexokinase and the rate of incorporation of [(14)C]-glucose into glycogen are much lower in liver from postnatal sheep than in rat liver. 3. The activities of hexose diphosphatase and glucose 6-phosphatase and the rates of incorporation of [(14)C]pyruvate and [(14)C]propionate into glycogen increase from low levels in the liver of foetal sheep to maxima a few weeks after birth. The activities in the liver of adult sheep are slightly lower. 4. The incorporation rate of [(14)C]pyruvate into glucose has been measured in liver slices from rats, sheep and chick embryos at several ages of these animals. This pathway is active in liver from foetal sheep, embryonic chicks and postnatal rats or sheep, but is absent from the liver from foetal rats. 5. Fructose metabolism, as measured by the rates of incorporation of [(14)C]fructose into glycogen and glucose in liver slices and by assays of liver ketohexokinase, is barely detectable in the liver of foetal sheep and appears soon after birth. 6. During development of the sheep, the incorporation rate of [(14)C]galactose into glycogen in liver slices is highest in foetal sheep and decreases with increasing age of the animal. 7. These findings are discussed with reference to the changing pattern of carbohydrate metabolism during neonatal development of liver in the sheep.

Animals↗
PubMed2001

Social facilitation of predatory, sheep-chasing behaviour in Norwegian Elkhounds, grey.

Sheep grazing on unfenced mountain pasture may be attacked by loose dogs, which may chase and kill sheep and separate lambs from their mothers. We have earlier shown that testing dogs individually towards sheep with an electronic dog collar may effectively reduce the chasing propensity 1 year later. The aim of this study was to investigate whether and how a non-chasing and a chasing companion dog influence sheep chasing in test dogs (20 Norwegian Elkhounds, grey), and whether this varies with sex or age. The test dogs' predatory behaviour towards sheep was examined by observation of the dogs in a fenced enclosure with sheep in 5min tests together with, first, a non-chasing (Hamilton stoever, a hound breed) and 2-3 days later a chasing (Border collie) companion dog. Physical contact with sheep was prevented. Initially, the test dogs exhibited a higher chasing motivation towards sheep in tests with a chasing, as compared with a non-chasing, companion (P<0.001). During the entire test, 60% of the dogs attacked sheep when accompanied by the non-chaser. All dogs attacked sheep when the chasing companion was present (P=0.008), although in only 8 of 20 cases the companion dog chased simultaneously with the test dog. In the chasing-companion tests, the attack latency was shorter (P<0.001) than in tests with a non-chasing companion. The attack severity was higher when attacking alone than when attacking together with the chasing companion (P=0.033). In these tests, the higher the attack severity, the shorter was the attack latency (P=0.006). A sequence analysis on chasing-companion tests showed that test dogs generally started with observing or showing interest in sheep, followed by attacks, which increased in severity. No sex differences were observed. In non-chasing companion tests, dogs aged up to 2.5 years exhibited a weaker initial hunting motivation than older dogs (P=0.025), but during the entire test were more inclined to attack (P=0.019). Taken together, our findings indicate that a companion dog showing intentions of predatory behaviour stimulates predatory chase in another dog, while a non-chasing companion has a limited influence on this. In tests certifying dogs for their refrainment of chasing sheep, well trained Border collies approaching sheep on command might be used to reveal the full predatory potential of the dog being tested.

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