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Water and electrolyte homeostasis in sheep without functional colons.

The regulatory role of the colon in water and electrolyte balance and the renal compensation which follows impairment of colonic function were assessed using sheep with ileorectal anastomosis (ILRAN sheep) on restricted and free water intake as experimental models. Faecal electrolyte loss sustained by the ILRAN group was eight to nine times greater than that in the control animals. When water was available ad lib., ILRAN sheep lost 2.81 and 0.51 more water per day via the faeces and urine, respectively, than the controls. Urine volume in the ILRAN sheep comprised largely electrolyte-free water and the renal retention of water was entirely secondary to the high degree of sodium reabsorption in these animals compared with the controls. On restricted water intake, the urine volume of the ILRAN sheep declined due to retention of electrolyte-free water and even greater absorption of sodium (and hence water by osmosis) by the kidney tubules. The latter observation was substantiated by a decrease in fractional excretion of sodium from 0.29 to 0.08% when water intake was restricted. Plasma aldosterone concentration was markedly elevated in the ILRAN sheep as a result of the excessive loss of sodium and water via the faeces. Activation of the renin-angiotensin-aldosterone sequence is believed to underlie the increased sensation of thirst (ILRAN sheep drank on average about 2.51 more water per day than the controls), the homeostatic response by the kidneys and the relatively lower plasma potassium in the ILRAN sheep compared to controls.

Animals↗

Single-dose pharmacokinetics of ampicillin/sulbactam (2:1) combination after intravenous administration to sheep and goats.

The pharmacokinetic behaviour of a combination of ampicillin and sulbactam (2:1) in six sheep and six goats after single intravenous doses of 20 mg kg body weight-1 (13.33 mg kg-1 of ampicillin and 6.67 mg kg-1 of sulbactam) was investigated by using a high-performance liquid chromatographic method for determining plasma concentrations. The objective was to determine whether there are differences between sheep and goats in the disposition kinetics of ampicillin and sulbactam. The plasma concentration-time curves were analysed by compartmental pharmacokinetic and non-compartmental methods. The disposition curves for both drugs were best described by a biexponential equation (two-compartment open model) in both sheep and goats. The mean (SD) elimination half-lives of ampicillin were 0.32 (0.05) h in sheep and 0.32 (0.04) h in goats, and the half-lives of sulbactam were 0.74 (0.10) h and 0.79 (0.18) h in sheep and goats, respectively. The apparent volumes of distribution of ampicillin and sulbactam were similar in the two species. Mean (SD) body clearances of ampicillin were 0.69 (0.07) litre h-1 kg-1 in sheep and 0.72 (0.11) litre h-1 kg-1 in goats, and the body clearances of sulbactam were 0.38 (0.03) and 0.38 (0.07) litre h-1 kg-1 in sheep and goats, respectively. There were no significant differences between any of the pharmacokinetic parameters of ampicillin and sulbactam in the sheep and goats.

Ampicillin↗

Sheep cytosolic branched-chain amino acid aminotransferase: cDNA cloning, primary structure and molecular modelling and its unique expression in muscles.

This paper presents the cloning and the molecular modelling of the cytosolic branched-chain amino acid aminotransferase (BCATc) from sheep brain. The sheep BCATc cDNA (3 kb) encodes a mature polypeptide of 385 amino acids with a calculated molecular mass of 43072.93 Da. The sequence of the sheep BCATc cDNA is more similar to other mammalian BCATc cDNAs (53-87% identical) than to the sheep mitochondrial branched-chain amino acid aminotransferase (52%). Sheep BCATc belongs to the IV Folding class of pyridoxal-5'-phosphate-depending enzymes. Based on the known structure of the branched-chain amino acid aminotransferase (BCAT) from Escherichia coli, a molecular model of sheep BCATc (amino acid residues 62-385) was built. This is the first three-dimensional model of any mammalian BCAT. It suggests that the enzymatic mechanism of sheep BCATc and likely of all mammalian BCAT is very similar to the mechanism of the E. coli BCAT and confirms the hypotheses regarding to the substrate binding sites of E. coli BCAT. Sheep skeletal muscle, which is the main in vivo site for transamination of branched-chain amino acids, exhibits an unique expression of BCATc.

Amino Acid Sequence↗

A case-control study on scrapie in Norwegian sheep flocks.

Scrapie first was detected in indigenous sheep in Norway in 1981, and from 1995 to 1997 an increase in the number of flocks with scrapie cases was recorded. These flocks were mainly in one geographical region. A study to identify risk factors for scrapie was conducted. The study had three frequency-matched controls selected for every case within the same Veterinary District. A questionnaire was submitted to 176 sheep flocks (42 had been scrapie flocks). The data obtained by the questionnaire were linked to data collected from governmental and industry registers. After imputing missing data using single random imputation, the statistical analysis was performed using multivariable conditional logistic regression. Purchase of female sheep from scrapie flocks, sharing of rams, or sharing of pastures between different flocks were the risk factors associated with the occurrence of scrapie. Of factors potentially sustaining and promoting the infection in the flock, number of winter-fed sheep, number of buildings for housing sheep, rams and ewes shared room during mating period and increase in the flock size were associated with scrapie. We interpret these findings to show that factors involving transfer of sheep between flocks or direct contact between sheep of different flocks are important for the spread of scrapie. Management factors are important for the development of scrapie. However, it was not possible to discriminate between the different management factors in this study at the flock level. Also, factors indicating awareness and interest of the farmer (as well as willingness to contact a veterinarian for diseased sheep) were related to the detection of scrapie in the flock.

Animal Husbandry↗

Molecular screening of sheep for bovine spongiform encephalopathy.

Bovine spongiform encephalopathy (BSE) may have transmitted to sheep through feed and pose a risk to human health. Sheep BSE cannot be clinically distinguished from scrapie, and conventional strain typing would be impractical on a significant scale. As human prion strains can be distinguished by differences in prion protein (PrPsc) conformation and glycosylation we have applied PrP(Sc) typing to sheep. We found multiple Western blot patterns of PrP(Sc) in scrapie, consistent with the known scrapie strain diversity in sheep. Sheep passaged BSE showed a PrP(Sc) banding pattern similar to BSE passaged in other species [Collinge, J., Sidle, K.C.L., Meads, J., Ironside, J. and Hill, A.F., Nature, 383 (1996) 685-690], both in terms of fragment size following proteinase K cleavage and abundance of diglycosylated PrP. However, none of the historical or contemporary scrapie cases studied had a PrP(Sc) type identical to sheep BSE. While more extensive studies, including sheep of all PrP genotypes, will be required to fully evaluate these findings, these results suggest that large scale screening of sheep for BSE may be possible.

Animals↗

Localization and genomic organization of sheep antimicrobial peptide genes.

Antimicrobial peptides are an abundant and diverse component of animal innate immunity. Within mammalian species, defensins and cathelicidins are the two principal antimicrobial peptide families. We identified and sequenced ten new sheep genes which encode potential antimicrobial peptides including two beta-defensins and eight cathelicidins. We mapped the two-exon beta-defensin genes to sheep chromosome 26 and the four-exon cathelicidin genes to sheep chromosome 19 using sheep-hamster somatic cell hybrids in conjunction with flow-sorted sheep chromosomes. These assignments confirm homology between sheep, cattle, mouse, and human antimicrobial peptide gene families. Contig construction for the sheep cathelicidin gene family demonstrates that three genes, OaDodeA, OaDodeB, and OaMAP-34, are present head-to-tail in a 14.5 kb region, and that four proline/arginine-rich genes, OaBac5, OaBac7.5, OaBac11, and OaBac6, are arranged head-to-tail in a region covering 30.5 kb. This richly diverse family of sheep cathelicidin peptides is encoded in a gene array which may reflect the mechanism of its evolution.

Amino Acid Sequence↗

Beneficial hemodynamic, endocrine, and renal effects of urocortin in experimental heart failure: comparison with normal sheep.

OBJECTIVES: The goal of this study was to determine the bioactivity of urocortin (Ucn) in experimental heart failure (HF). BACKGROUND: Urocortin may participate in cardiovascular function and pressure/volume homeostasis. Its effects in HF are unknown. METHODS: Eight normal sheep and eight sheep with pacing-induced HF received ovine Ucn (10, 50, and 100 mg intravenous boluses at 2-h intervals) in vehicle-controlled studies. RESULTS: Urocortin boluses dose-dependently increased plasma Ucn (p < 0.001). Pharmacokinetics were similar in normal and HF sheep with half-lives approximating 1.3 and 19.5 h for the first and second phases, respectively. In HF, cardiac output increased (twofold), while peripheral resistance, left atrial pressure (both 50% falls: p < 0.001), and mean arterial pressure (p < 0.05) fell. In normal sheep, changes in peripheral resistance and atrial pressure were blunted and in arterial pressure were directionally opposite. Urocortin induced persistent, dose-dependent falls (30% to 50%) in plasma vasopressin, renin activity, aldosterone, natriuretic peptides (all p < 0.001), and endothelin-1 (p < 0.05) in HF sheep, while adrenocorticotrophic hormone and cortisol levels rose acutely (both p < 0.001). In comparison, Ucn in normal sheep resulted in a similar rise in cortisol and fall in aldosterone, no significant effects on plasma renin activity and natriuretic peptides, and a rise in vasopressin. Urocortin produced dose-dependent, sustained increases in urine volume (twofold, p < 0.01), sodium excretion (>9-fold rise, p < 0.001), and creatinine clearance (p < 0.001) in HF sheep. No significant renal effects were observed in normal sheep. CONCLUSIONS: Urocortin has profound and sustained hemodynamic, hormonal, and renal effects in experimental HF. Urocortin may have a role in pressure/volume homeostasis in HF and may provide a novel therapeutic approach to this disease.

Animals↗

Specific frequency distribution of erythrocytic X-protein alleles in indigenous sheep populations in East Asia.

The nonhemoglobin erythrocytic X-protein polymorphism consisting of two phenotypes called X-positive [X(+)] and X-negative [X(-)] was determined in 576 unrelated healthy native sheep of East Asia, using one-dimensional and horizontal starch gel electrophoresis. A striking difference in the frequency of the X allele coding dominantly for the X(+) type between the northern and southern populations of native East Asian sheep divided by the boundary of the Himalaya Mountains was seen (P < 0.0001). The X allele frequency ranged from 0 to 0.0438 with an average of 0.0323 in the northern population examined, consisting of the Bhyanglung, Baruwal, Yunnan, and Khalkhas sheep belonging to the Tibetan and Mongolian sheep groups. In contrast, the frequency of the same allele was in the range of 0.2037-0.4655 and the mean frequency was 0.2998 in the southern population tested, consisting of the Bengal, Kagi, Lampuchhre, Vietnamese, and Myanmar sheep, which belong to the Indian sheep group. This finding suggests that the X allele appears to be an Indian sheep marker and is potentially important in phylogenetic studies on native sheep populations, especially in East Asia.

Alleles↗

Apolipoprotein E polymorphism and plasma lipid levels in Native Mongolian sheep.

Apolipoprotein E (apoE) phenotypes were determined in 199 unrelated native sheep (Khalkhas line) of Central Mongolia, using a polyacrylamide gel isoelectric focusing-immunoblotting technique, and the plasma lipid levels in different phenotypes were assayed enzymatically. Twenty-eight phenotypes were identified in this sheep. In addition to all the previously detected seven apoE variants composing the phenotypes, four new variants were discovered, which were called E8, E9, E10, and E11. From the population data, these were found to be genetically controlled by four codominant alleles, designated APOE8, APOE9, APOE10, and APOE11, based on the same mode of inheritance as in the seven variants. These alleles were detected at a low frequency, in the range of 0.005 to 0.0126. The Khalkhas sheep differed most significantly from the Baruwal and Lampuchhre sheep of Nepal and the Vietnamese sheep with respect to the allele frequencies found in some Asian local sheep previously examined. Type 1/1 and/or 2/7 sheep had significantly higher plasma levels of total cholesterol and low-density lipoprotein cholesterol than type 7/7 sheep (P < 0.05 and/or P < 0.02).

Animals↗

Characterization of indigenous fat-tailed and fat-rumped hair sheep in Kenya: diversity in blood proteins.

This paper reports the variation in five blood proteins from five populations of sheep found in Kenya. Blood samples were collected from a total of 309 adult sheep of both sexes in Kwale, Makueni and Kakamega districts for the fat-tailed sheep and in Isiolo district for the fat-rumped hair sheep. Fine-wooled Merino sheep were used in this study as the reference population. Transferrin, esterase-A and esterase-C were polymorphic in all the populations investigated, while albumin was monomorphic for the S allele in the fat-tailed sheep and haemoglobin was fixed for the B allele in the Kwale, Makueni and Isiolo populations. Phylogenies derived from the pairwise genetic distance estimates showed a clear separation between the indigenous sheep populations and the exotic Merino. However, the topology of the former showed rather poor consistency with their morphological classification based on the localization of their fat deposits, namely fat-tailed or fat-rumped hair sheep.

Alleles↗

A short-term study in sheep with the Groningen temporomandibular joint prosthesis.

As part of the pre-clinical testing process of a newly developed temporomandibular joint (TMJ) prosthesis, animal experiments were performed. In 14 sheep, the right TMJ was replaced by the developed TMJ prosthesis. The prosthesis consisted of a skull part, a mandibular part and an intervening polyethylene disc. In the first series (6 sheep), three designs were tested, differing in the applied metal (stainless steel or titanium) and in the fitting method of the skull part (a fitting member or bone cement). The sheep were sacrificed after 8-16 weeks. In the second series (8 sheep), the preferred titanium fitting member design was applied, and the sheep were sacrificed after 2-10 weeks. One sheep was excluded because no correct position of the prosthesis parts could be achieved. At sacrifice, the removal torque of the screws was measured, and the surrounding tissues were harvested for histologic examination. The sheep recovered well and functioned until the end of the scheduled sacrifice date. Encountered problems were two disc dislocations, one fistula formation, and one screw failure. All mandibular parts were clinically stable, as were most skull parts with a fitting member, and one of both skull parts fitted with bone cement. The clinically observed stability was confirmed by the removal torque values, which indicated well-integrated screws. It is concluded that the TMJ prosthesis could remain stable and functional over the initial healing period. The main restriction of the sheep model is the much larger translatory capacity compared with patients, which adversely influences tissue healing.

Animals↗

Lymphocyte subpopulations in sheep during the early stage of experimental infection with bovine leukaemia virus.

Sheep were experimentally infected with bovine leukaemia virus (BLV) by the inoculation of PBL from leukaemic sheep. Antibodies to viral structural proteins were detected at from 2 to 6 weeks after inoculation. At seroconversion, all sheep had a marked increase in the number of circulating lymphocytes, due essentially to an increase in the number of B cells. The number of circulating B cells then decreased but remained higher than pre-infection levels. A second increase in this population preceded the development of a B cell lymphoblastic leukaemia. Generalized lymphosarcoma was diagnosed at necropsy of all sheep. Variation between individual sheep in the time from infection to the development of tumours allowed two clearly delineated groups of nine sheep to be compared. A study of changes in the B cell and T cell populations during the first 16 weeks of infection suggested that the initial response to infection influences the subsequent rate of leukaemogenesis. At seroconversion the number of circulating B cells was significantly higher in group 1 (10.16 +/- 1.51 X 10(9)/l) than in group 2 (6.47 +/- 2.76 X 10(9)/l). Group 1 sheep became leukaemic at 20-50 weeks after infection, whereas group 2 sheep did not do so until 70-95 weeks after infection.

Animals↗

Utilization of free fatty acids by starved and pregnant sheep.

1. The rate of entry into the plasma of stearic acid in fed and starved non-pregnant sheep and of palmitic acid in fed and starved pregnant sheep has been measured by a continuous-infusion isotope-dilution method. 2. In non-pregnant sheep the entry rate of stearic acid rose from 0.38mg./min./kg. when fed to 0.69mg./min./kg. after 72hr. starvation. In pregnant sheep, the entry rate of palmitic acid rose from 0.55mg./min./kg. when fed to 0.64mg./min./kg. on starvation. 3. The entry rates of palmitic acid and stearic acid are related to their respective plasma concentrations. 4. At a given plasma concentration the entry rate of palmitic acid in pregnant sheep was greater than that of stearic acid in non-pregnant sheep. 5. There was no detectable conversion of palmitate or stearate into other plasma long-chain fatty acids. There was negligible incorporation of fatty acids into other plasma lipids with the exception of the plasma triglycerides of fed pregnant sheep. 6. Up to 12% of expired carbon dioxide was derived from palmitic acid or stearic acid. The high rate of oxidation of plasma palmitic acid in fed pregnant sheep is noteworthy.

Animals↗

Regulation of carnitine palmitoyltransferase activity by malonyl-CoA in mitochondria from sheep liver, a tissue with a low capacity for fatty acid synthesis.

The characteristics of inhibition of carnitine palmitoyltransferase (CPT) I by malonyl-CoA were studied for the enzyme in mitochondria isolated from sheep liver, a tissue with a very low rate of fatty acid synthesis. Malonyl-CoA was as potent in inhibiting the sheep liver enzyme as in inhibiting the enzyme in rat liver mitochondria. CPT I in guinea-pig liver mitochondria was also similarly inhibited. The inhibition showed the same time-dependent characteristics previously established for the rat liver enzyme. Methylmalonyl-CoA was as effective an inhibitor of CPT I as malonyl-CoA in sheep liver mitochondria, but did not affect CPT I activity in mitochondria from rat or guinea-pig liver. The concentrations of malonyl-CoA required to inhibit CPT I in sheep liver mitochondria in vitro were similar to those found in freeze-clamped sheep liver samples (about 7 nmol of malonyl-CoA/g wet wt.). In sheep liver cells the content of malonyl-CoA was only one-tenth of that observed in vivo when glucose only was added to the incubation medium. Inclusion of acetate and/or insulin increased the malonyl-CoA content about 10-fold, to values similar to those observed in vivo. The rate of fatty acid synthesis in sheep liver cells was about 1% of that observed in rat liver, but was correlated with the concentrations of malonyl-CoA in the cells under various incubation conditions. These observations are discussed in relation to (i) the regulatory role of malonyl-CoA in tissues that have a low capacity for fatty acid synthesis, and (ii) the utilization by sheep liver of propionate as a gluconeogenic precursor.

Acyl Coenzyme A↗

Animal host associated differences in Shiga toxin-producing Escherichia coli isolated from sheep and cattle on the same farm.

AIMS: To investigate if cattle on the same farm as sheep are a possible risk factor for stx in sheep and to determine whether or not sheep and cattle on the same farm share the same stx pool. METHODS AND RESULTS: Faecal samples from sheep and cattle were screened for stx by polymerase chain reaction (PCR). Of these samples, 87.6 and 64.6% were stx positive in sheep and cattle, respectively. There was no difference in stx occurrence in sheep from farms with or without cattle. From stx positive samples, 118 Shiga toxin-producing Escherichia coli (STEC) isolates were recovered by a filter-hybridization method. Serotyping, PCR and pulsed-field gel electrophoresis (PFGE) showed that there was a distinct association between serotypes, stx profiles and animal species. CONCLUSIONS: Keeping animals together in pens, which enhances faecal-oral contact, is suggested as a possible explanation for the differences seen in stx occurrence. Sheep and cattle isolates are distinctly different in serotype and stx profile although isolated from the same farm, and are more related to isolates within the same serotype with the same stx profile than to isolates with different serotype from the same farm. SIGNIFICANCE AND IMPACT OF STUDY: The study supports the animal-host relationship hypothesis suggested in other studies and indicates that the STEC sheep reservoir in Norway may not pose a serious public health risk.

Animal Husbandry↗

Modulation of Na+ transport across isolated rumen epithelium by short-chain fatty acids in hay- and concentrate-fed sheep.

The effect of increasing concentrations of short-chain fatty acids [SCFA; mixture of the Na+ salts of acetic acid (62.5%), propionic acid (25.0%) and of butyric acid (12.5%)] on Na+ transport of sheep rumen epithelium was studied in vitro. The conventional Ussing chamber method was used for measuring Na+ transport rates (22Na+), short-circuit current (Isc) and tissue conductance (GT) of isolated rumen epithelium. SCFA in the buffer solution on the mucosal side caused a linear increase of Jnet Na+ from 1.14, to 1.22, 1.78 and 2.50 microeq/cm2/h in hay-fed sheep at 0, 15, 40 and 80 mmol/l SCFA, respectively. In a second study, the effect of higher SCFA concentrations [0 (control), 80, 100 and 120 mmol/l] was investigated with epithelia from two groups of sheep. One group was subjected to hay ad libitum, whereas the other received concentrate feed (800 g/day in equal portions at 7.00 am and 3.00 pm) and hay ad libitum. Epithelia from concentrate-fed sheep again showed a significant (p < 0.05) and linear increase in Jnet Na+ at 80, 100 and 120 mmol/l. However, in hay-fed sheep, the difference in increase among 80, 100 and 120 mmol/l SCFA was not significant, indicating that, above 80 mmol/l SCFA Jms and Jnet exhibit saturation. Moreover, Na+ fluxes (Jms and Jnet) were generally higher in concentrate-fed than in hay-fed sheep at all SCFA concentrations and significant differences were observed at 100 and 120 mmol/l SCFA. The obtained results confirm the effect of SCFA on Na+ transport and are in agreement with studies regarding feeding regimes and electrolyte transport in the rumen. The important new observation is the increase of Na+ transport in concentrate-fed sheep even at high concentrations of SCFA (100 and 120 mmol/l). The enhanced activity of the Na+/H+ exchanger at these SCFA concentrations supports the assumption that the capacity for regulating the intracellular pH by extrusion of protons is increased, suggesting an adaptation in concentrate-fed sheep. This adaptation could prevent possible disturbances of epithelial functions (transport and barrier) under conditions of increased SCFA absorption.

Absorption↗

Changes in behaviour, cortisol and lymphocyte types during isolation and group confinement of sheep.

This experiment compared changes in complex behaviour patterns, adrenal corticosteroid secretion and the numbers of various types of lymphocytes in sheep that were subjected to the stress of confinement. Grazing Merino ewes (n=80 in five replicated experiments) were confined either in groups of four per pen or in total isolation from other sheep. The percentage of CD4+ lymphocytes increased while the percentage of CD8+ lymphocytes decreased over the experimental period. This result was more pronounced in isolated sheep than in grouped sheep. The increase in CD4:CD8 was greater for isolated sheep than for grouped sheep and greater for 2 week sheep than for 3 week sheep. The percentage of CD5+ cells also increased, less so in isolated than in grouped animals. Interpreting these changes as a recovery of immune competence following introduction of a stressor, it is apparent that isolation impaired immune system recovery more severely than group confinement. Physiological and behavioural adaptation over the period were characterized by a decline in the adrenocortical response, resumption of the normal pattern of flocking behaviour and a reduction in motor activity during the test. These findings add to the evidence pointing to the possible correspondence between critical features of the psychoneural, neuroendocrine and immune systems.

Animals↗

Autoaugmentation peritoneocystoplasty in a sheep model.

OBJECTIVE: To report our experience with autoaugmentation peritoneocystoplasty (AAPC) in a sheep model, and to compare the results with autoaugmentation gastrocystoplasty (AAGC) in a sheep model and in paediatric patients. MATERIALS AND METHODS: Ten 6-month-old male lambs underwent bladder augmentation by detrusorotomy. A flap of parietal peritoneum, dissected from the anterior abdominal wall, was used to cover the bladder mucosa. The sheep were evaluated by urodynamics 6 months after surgery. Bladder compliance (bladder volume/intravesical pressure) was calculated for the bladder capacity at leakage. The urodynamic results were compared with age-matched control sheep and with 12 sheep that had undergone AAGC; the results were assessed using the Mann-Whitney U-test. RESULTS: In two of the 10 sheep, bladder volumes after AAPC increased by > 100%, although for the group, the mean (range) bladder volume after augmentation, at 159 (42-261) mL, was not significantly different from that before surgery (mean 143 mL). Bladder volumes after AAPC were not significantly different from those in the control sheep (mean 205 mL) but were significantly less than in the AAGC group (mean 317 mL; P < 0.05). Bladder compliance at leak capacity in the AAPC group (mean 5.4 mL/cmH2O) was also not significantly different from the controls (mean 9.1 mL/cmH2O), but was lower than the in the AAGC animals (median 14.6 mL/cmH2O; P < 0.05). CONCLUSIONS: AAPC in a sheep model does not result in a reliable increase in bladder volume or compliance. The volume and compliance are inferior to those found in bladders augmented by AAGC.

Animals↗