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Stimulation of the titre of a sheep serum factor that is related to the sheep complement system.

Serum from most sheep subjected to a single jugular bleeding, repeated bleeding or an intraperitoneal injection of yeast and repeated bleeding, showed an increase in the titre of a serum component called serum factor. Serum factor reached a peak titre 2 to 5 days after treatment started. For some sheep, the titre was elevated for a 9- to 12-day period whereas for others the titre dropped markedly on day 7 followed by a rise on day 9. Serum factor reacts with sheep erythrocytes sensitised with rabbit antibody (sheep E-rabbit A). Serum factor can be detected on sheep E-rabbit A using guinea-pig antiserum that reacts with sheep complement (C). Serum factor is inactivated by heating at 56 degrees C for 30 minutes, but is only partially inactivated at 50 degrees C for 30 minutes. The reaction of serum factor with sheep E-rabbit A is inhibited by chelators of Ca++ and/or Mg++. Serum factor appears to be related to the sheep C system. Preliminary results suggest it may he a component of the classical pathway of sheep C, possibly the second component, C2.

Journal Article↗

The growth of sheep scab lesions in relation to sheep breed and time of the year.

Numerically equal groups of between five and ten young Merino and Dorper sheep were artificially infested with sheep scab mites, Psoroptes ovis. Separate groups of sheep were infested during the early winter of two successive years as well as during the early summer of the second year. Lesion growth was measured at two-weekly intervals for a period of six weeks during the first winter and for eight weeks during the second winter and in the summer. The rate of lesion growth and mean lesion size on Merino sheep was significantly greater than that on Dorper sheep at each two-weekly recording during the winter of both years. The rate of growth of the lesions did not differ significantly between the two sheep breeds during summer, although, with the exception of eight weeks post-infestation, the mean lesion size on Merino sheep was significantly larger than that on Dorper sheep at each assessment.

Animals↗

The rate of spread of sheep scab within small groups of Merino and Dorper sheep.

A single Merino sheep, artificially infested with the sheep scab mite, Psoroptes ovis, and a similarly infested Dorper sheep were placed with 9 uninfested Merino or 9 uninfested Dorper sheep respectively during winter and the rate of spread of infestation on the uninfested sheep observed. The same procedure was repeated in summer. It took 14 and 8 weeks respectively in winter before all sheep in the 2 groups displayed lesions of sheep scab, whereas in summer it took 10 and 12 weeks before all sheep had lesions.

Animals↗

Variations in sheep serum conglutinating and haemolytic activity for sheep erythrocytes sensitized by rabbit antibody.

Based on conglutinating and haemolytic reactions with sheep erythrocytes (E) sensitized by rabbit antibody (A), three types of sheep sera were encountered. Type 1 sera do not conglutinate or haemolyse sheep E-rabbit A. Type 2 sera failed to conglutinate or haemolytically active. Type 3 sera have both activities. Serum from one type 1 sheep still failed to conglutinate 5 days after venepuncture but was not haemolytically active (i.e., type 2). Some sheep that initially had type 2 sera had, five days after an intraperitoneal injection of yeast cells, sera with conglutinating activity (type 3 sera). Type 1, 2 and 3 sera all had haemolytic activity with human E-sheep A indicator cells. Pooled type 3 sera have the highest conglutinating titres with sheep E-rabbit A after 10 min incubation at 39 degrees C. At this stage, the haemolytic titres were very low. From 10 min, the conglutinating titres decreased whereas the haemolytic titres gradually increased until 80 min. Optimal conglutinating activity required less rabbit A to sensitize sheep E than did haemolytic activity.

Animals↗

The sheep erythropoietin gene: molecular cloning and effect of hemorrhage on plasma erythropoietin and renal/liver messenger RNA in adult sheep.

Erythropoietin (Epo) production was studied in adult sheep. Nine ewes, body weight (BW) 39 +/- 1.3 kg, were hemorrhaged a volume of blood equivalent to 1.6% BW, and sampled at 0, 2, 4, 6, 24 h. Oxygen content (O2 CT) decreased by 2.7 +/- 0.6 ml/dl at 2 h. Plasma immunoreactive (IR) Epo was only significantly increased at 24 h, from 18.5 +/- 3.5 to 40 +/- 10.7 mU/ml (mean +/- SEM). A further 5 ewes were bled extensively (2793 +/- 82 ml) over 54 h, and killed for Epo mRNA determination. The O2 CT decreased from 12.3 +/- 1.6 to 4.1 +/- 0.6 ml/dl, and plasma Epo increased from 15 +/- 4 to 1675 +/- 287 mU/ml. The sequence of ovine Epo cDNA was derived from the kidney RNA of a severely bled sheep using reverse transcription/polymerase chain reaction (RT/PCR), and from an ovine Epo genomic clone. The cDNA encodes a peptide of 194 amino acids, including a 27 amino acid signal peptide. The deduced amino acid sequence of sheep Epo shows 82%, 78% and 80% homology with mature Epos of human, mouse and monkey, respectively. The gene structure resembles closely those of human and mouse, with 5 exons and 4 introns. The expression of the ovine Epo gene in tissues from normal and hemorrhaged sheep was analysed by a competitive RT/PCR method. Epo mRNA was difficult to detect in liver from normal sheep, but was detectable at 0.01-0.04 amol/microgram total RNA in kidney from normal sheep. In the kidneys of severely bled sheep, the Epo mRNA levels (per micrograms total RNA) increased 400-1500-fold compared to that of normal kidneys, and were approximately 60-fold greater than those in the livers of the hemorrhaged sheep.

Amino Acid Sequence↗

Oxidative metabolism of long-chain fatty acids in mitochondria from sheep and rat liver. Evidence that sheep conserve linoleate by limiting its oxidation.

Mitochondria isolated from the livers of sheep and rats were shown to oxidize palmitate, oleate and linoleate in a tightly coupled manner, by monitoring the oxygen consumption associated with the degradation of these acids in the presence of 2mM-L-malate. Rat liver mitochondria oxidized linoleate and oleate at a rate 1.2-1.8 times that of palmitate. Sheep liver mitochondria had a specific activity for the oxidation of palmitate that was 50-80% of that of rats and a specific activity for the oxidation of oleate and linoleate that was 30-40% that of rats. This would indicate that sheep conserved linoleate by limiting its oxidation. Carnitine acyltransferase I (CAT I) actively esterified palmitoyl-CoA and linoleate to carnitine in both rat and sheep liver mitochondria, and in both cases the rate for linoleate was faster than for palmitate. The CAT I reaction in both rat and sheep liver was inhibited by micromolar amounts of malonyl-CoA. With 90 microM-palmitoyl-CoA as substrate, CAT I was inhibited by 50% with 2.5 microM-malonyl-CoA in rats, and in sheep, 50% inhibition was found with all malonyl-CoA concentrations tested (1-5 microM). With 90 microM-linoleate as substrate for CAT I, a much larger difference in response to malonyl-CoA was seen, the rat enzyme being 50% inhibited at 22 microM-malonyl-CoA, whereas sheep liver CAT I was 91% and 98% inhibited at 1 microM- and 5 microM-malonyl-CoA respectively. We propose that malonyl-CoA may act as an important regulator of beta-oxidation in sheep, discriminating against the use of linoleate as an energy-yielding substrate.

Animals↗

The comparative sensitivity of sheep and chicken embryos to bluetongue virus and observations on viraemia in experimentally infected sheep.

The virus titre in sheep blood samples received from BT-suspected cases in the field was assayed in sheep and in chicken embryos. These infected blood samples represented 3 different BT virus types: 4, 10 and 16. Three identical experiments were performed, one with each of the 3 different virus types. Ten-fold dilutions of the infected blood samples were prepared and 1 ml of each blood dilution was inoculated IV into series of 10 to 12-month old susceptible sheep; at the same time 0.1 ml of each dilution was inoculated IV into series of 10 to 13-day-old chicken embryos. The virus titre was found to be similar when assayed in the two host systems. There was no correlation between the amount of virus inoculated and the severity of symptoms in the inoculated sheep. The virus content in daily blood samples collected from the experimental sheep was assayed by IV inoculation of CE. Virus was isolated from all the reacting sheep and was detected sometimes as early as 1 day PI and as late as 30 days PI. A high titre of log10 4.0 to 7.0 per 1 ml of blood was recorded during several consecutive days before and after the onset of clinical signs. There seemed to be an inverse ratio between the amount of virus inoculated and the number of days the virus persisted in the bloodstream. The neutralisation index in day 22 serum samples was 3.5 to 4.5. Virus was isolated from some of the reacting sheep on the day that these antibody levels were recorded. Since the comparative simultaneous titrations of BT virus in sheep and in CE yielded similar results, the IV inoculation of CE is advocated as the routine method to be employed for laboratory diagnosis of this disease.

Animals↗

Experimental transmission of sheep scrapie by intracerebral and oral routes to genetically susceptible Suffolk sheep in the United States.

Scrapie is a naturally occurring fatal neurodegenerative disease of sheep and goats. Susceptibility to the disease is partly dependent on the genetic makeup of the host. This study documents clinicopathological findings and the distribution of abnormal prion proteins (PrPres) by immunohistochemical and Western blot techniques, in tissues of genetically susceptible sheep inoculated with US sheep scrapie agents. Four-month-old Suffolk lambs (QQ or HQ at codon 171) were inoculated (5 intracerebrally and 19 orally) with an inoculum (#13-7) consisting of a pool of scrapie-affected sheep brains. Intracerebrally inoculated animals were euthanized when advanced clinical signs of scrapie were observed. Orally inoculated animals were euthanized at predetermined time points (4, 9, 12, 15, and 21 months postinoculation [PI]) and thereafter when the animals had terminal signs of disease. All intracerebrally inoculated animals exhibited clinical signs of scrapie and were euthanized between 13 and 24 months PI. Spongiform lesions in the brains and PrPres deposits in central nervous system and lymphoid tissues were present in these sheep. In orally inoculated sheep, clinical signs of scrapie were seen between 27 and 43 months PI in 5/9 animals. The earliest detectable PrPres was observed in brainstem and lymphoid tissues of a clinically normal, orally inoculated sheep at 15 months PI. Three of the 4 clinically normal sheep were positive at 15, 20, and 49 months PI by PrPres immunohistochemistry.

Administration, Oral↗

Sheep serum complement sensitisation of sheep erythrocyte-rabbit antibody complexes for haemolysis by guinea-pig complement plus EDTA or Mg2+-EGTA.

Sheep erythrocyte (E)-rabbit antibody (A) complexes incubated with sheep serum diluted up to 1:5120 or 1:20480 and washed can be haemolysed by guinea-pig (g-p) serum (complement, C) containing EDTA or Mg2+-EGTA respectively as haemolytic finishing reagents. Sheep E carrying a high dose of rabbit A were necessary for this reaction, particularly with g-p C-EDTA. G-p serum (stored by freezing) was active as a haemolytic finishing reagent with both EDTA and Mg2+-EGTA. Reconstituted freeze-dried g-p serum (also stored by freezing) was haemolytically active with Mg2+-EGTA only. G-p serum preserved by Richardson's method did not function as a finishing reagent with EDTA or Mg2+-EGTA. A non-haemolytic prozone occurred with sheep E-rabbit A treated with dilutions of sheep serum or body fluid up to 1:160, particularly when g-p C (frozen)-EDTA was used as the finishing reagent. Sheep E-rabbit A were sensitized by serum, foetal lamb serum, pericardiac-, synovial- or ovarian follicle-fluids colostrum or milk for haemolysis by g-p C (frozen)-EDTA or -Mg2+-EGTA. With the C3 inhibitors cobra venom factor or salicylaldoxime, serum sensitisation of sheep E-rabbit A for haemolysis by g-p C (frozen)-EDTA or -Mg2+-EGTA was not blocked. Sensitisation by serum heated at 50 degrees C for 30 min (partial inactivation of C2) was incomplete. Inhibitors of C1 (antrypol, chelators of Ca2+ or heating serum at 56 degrees C for 30 min) partially or fully blocked sensitisation for haemolysis by both g-p C (frozen)-EDTA or -Mg2+-EGTA. These results show that at a minimum, components C1, C4 and C2 are present and functionally active in serum and some body fluids of sheep.

Animals↗

Effect of sheep and human follicular fluid on the maturation of sheep oocytes in vitro.

This study examines the effect of sheep and human follicular fluid on the in vitro maturation (IVM) of sheep follicular oocytes. Oocyte cumulus complexes recovered post mortem were matured for 24 to 26 h at 38.6 degrees C, 5% CO(2) in air, in TCM-199 bicarbonate medium supplemented with 20% fetal calf serum (FCS) and, where stated, with maturation hormones, including FSH (5.0 microg/ml), LH (5.0 microg/ml) and estradiol (1 microg/ml), or with sheep follicular fluid recovered from large (>5 mm) or small (2 to 5 mm) ovarian follicles post mortem, or with human periovular follicular fluid obtained during routine IVF procedures. The matured oocytes were then denuded, and their maturation stage and developmental capacity were assessed by in vitro fertilization (IVF) and culture (IVC). It was found that inclusion of sheep or human follicular fluid or hormone supplements in the IVM media more than doubled the number of oocytes completing maturation (FCS alone 33%, compared with 76.2% for maturation hormones, 84.2% for fluid from large and 69.6% for fluid from small sheep follicles and 82.6% for human follicular fluid), and significantly increased fertilization rates (FCS alone 51.6%, compared with 71.9% for maturation hormones, 78.4% for fluid from the large and 75.7% for fluid from small sheep follicles and 73.1% for human follicular fluid) without discernible adverse effects on the development of the cleaving embryos to the morula or blastocyst stage in culture. Omission of FCS and supplements from the IVM medium resulted in a marked reduction (56%) in the number of oocytes maturing. This reduction could be offset to a large part, but not completely, by inclusion of human follicular fluid or human follicular fluid plus LH (5 microg/ml) in the medium. The results of this study show that addition of sheep or human follicular fluid to maturation medium can enhance rather than inhibit the maturation and fertilizability of sheep follicular oocytes in vitro.

Journal Article↗

Phospholipid fatty acids in brains of normal sheep and sheep with ceroid-lipofuscinosis.

The ceroid-lipofuscinoses are a group of inherited diseases of humans and animals characterised by brain atrophy and the storage of a fluorescent lipopigment. Brain grey matter phospholipid fatty acids of diseased sheep are compared with those of normal sheep. Phosphatidylethanolamine of diseased sheep contains more 18:1(n-9) and less 22:6(n-3) than normal and their phosphatidylcholine less 16:0. Other differences are minor. All differences are in the same direction as those reported for the infantile form of human ceroid-lipofuscinosis, but are smaller. Normal sheep grey matter phosphatidylinositol contains 8.5% 20:4(n-6) and 24.6% 22:6(n-3), in contrast to 28.5 and 4.6%, respectively, in humans. The other sheep phospholipids have similar fatty acid profiles to those from humans. Apart from low levels of 20:3(n-9) and 22:3(n-9) they contain no additional non-essential fatty acid derived species. No sign of essential fatty acid deficiency occurs in either diseased or normal sheep. It is concluded that sheep must conserve their restricted essential fatty acid supply for structural functions, and that an abnormality in fatty acid metabolism is not primarily involved in the pathogenesis of ceroid-lipofuscinosis. The results also call into question the primary role of peroxidation of polyunsaturated fatty acids in lipopigment formation in this disease.

Animals↗

The greater susceptibility of North Ronaldsay sheep compared with Cambridge sheep to copper-induced oxidative stress, mitochondrial damage and hepatic stellate cell activation.

Sheep of the semi-feral North Ronaldsay (copper-sensitive) and domesticated Cambridge (copper-tolerant) breeds were compared in respect of pathological changes and protein expression in the liver as a result of excessive dietary copper. Acute mitochondrial damage and hepatic stellate cell (HSC) activation with collagen synthesis occurred in response to moderate copper overload in North Ronaldsay but not in Cambridge sheep. Mitochondrial degradative changes occurred either as ballooning degeneration and rupture with subsequent autophagic degradation or as mitochondrial matrical condensation (pyknosis). In North Ronaldsay sheep prolonged exposure to copper produced mitochondrial hyperplasia and hypertrophy, and nuclear damage with necrosis. Cytosolic isocitrate dehydrogenase (IDH), an enzyme responsive to oxidative stress, was induced in the liver of Cambridge sheep receiving a Cu-supplemented diet but was undetectable in the non-supplemented control sheep. Conversely, IDH was detected at similar levels in both control and copper-supplemented North Ronaldsay sheep, indicating a lower threshold response, and an enhanced susceptibility, to oxidative stress. "Upregulation" of mitochondrial thioredoxin-dependent peroxidase reductase (antioxidant protein-1) in the hepatic cytosol of the North Ronaldsay (but not Cambridge) sheep affirmed the increased susceptibility of the mitochondria to Cu-induced oxidative stress in this breed. Likewise the upregulation of cathepsin-D indicated increased lysosomal activity and HSC activation. The findings may be relevant to copper toxicosis in human infants.

Animals↗

Survival away from sheep and alternative methods of transmission of sheep lice (Bovicola ovis).

Transmission of sheep lice is thought to occur mainly by sheep to sheep contact although the possibility of other sources of infestation is often suggested. This study investigated the period of survival of Bovicola ovis after removal from sheep under varying conditions and assessed the likelihood of new infestations arising from contaminated facilities, wool caught on fences and shearers' footwear. In laboratory studies with lice held away from sheep at 4, 20, 25 and 36.5 degrees C, adults and nymphs survived longest at 25 degrees C (LT90 of 11.7 and 24.1 days for adults and large nymphs, respectively). Nymphs survived longer than adults and lice provided with raw wool survived longer than lice provided with wool that had been degreased. Nymphal lice survived for up to 29 days on unscoured wool at 36.5 degrees C, but the LT50 was less than 9 days in most experiments. In shearing sheds in winter and early spring lice survived for up to 14 and 16 days, respectively. These periods of survival are considerably longer than previously indicated for B. ovis. Most lice dropped out of wool staples attached to a fence within 1 h and only two of a total of 225 lice were still present after 24 h, suggesting that sheep are unlikely to become infested from wool caught on fences. Adult and nymphal lice readily transferred to shearers' moccasins and survived there for up to 10 days, indicating that transmission of lice on the footwear of shearers or other sheep handlers may be a cause of new infestations. Microwaving each moccasin for 5 min killed all lice and may provide a simple method of reducing the likelihood of transmission of B. ovis between properties.

Animal Husbandry↗

Molecular analysis of cases of Italian sheep scrapie and comparison with cases of bovine spongiform encephalopathy (BSE) and experimental BSE in sheep.

Concerns have been raised about the possibility that the bovine spongiform encephalopathy (BSE) agent could have been transmitted to sheep populations via contaminated feedstuffs. The objective of our study was to investigate the suitability of molecular strain typing methods as a surveillance tool for studying scrapie strain variations and for differentiating PrP(Sc) from sheep scrapie, BSE, and sheep BSE. We studied 38 Italian sheep scrapie cases from 13 outbreaks, along with a British scrapie case, an experimental ovine BSE, and 3 BSE cases, by analyzing the glycoform patterns and the apparent molecular masses of the nonglycosylated forms of semipurified, proteinase-treated PrP(Sc). Both criteria were able to clearly differentiate sheep scrapie from BSE and ovine experimental BSE. PrP(Sc) from BSE and sheep BSE showed a higher glycoform ratio and a lower molecular mass of the nonglycosylated form compared to scrapie PrP(Sc). Scrapie cases displayed homogeneous PrP(Sc) features regardless of breed, flock, and geographic origin. The glycoform patterns observed varied with the antibody used, but either a monoclonal antibody (MAb) (F99/97.6.1) or a polyclonal antibody (P7-7) was able to distinguish scrapie from BSE PrP(Sc). While more extensive surveys are needed to further corroborate these findings, our results suggest that large-scale molecular screening of sheep populations for BSE surveillance may be eventually possible.

Animals↗

An outbreak of sheep pox on a sheep breeding farm in Jammu, India.

An outbreak of sheep pox occurred in December 2001 on a sheep breeding farm in Jammu, India. The farm maintains three exotic breeds of sheep, i.e. American Merino, Rambouillet and Australian cross. The disease agent was confirmed as sheep pox virus by clinical and post-mortem examination as well as laboratory testing. Typical pock lesions were dispersed over the body of the affected animals with nodular lesions observed in the lung tissue of the dead animals. Sheep pox virus antigen and antibody were detected in infected tissue and convalescent sera, respectively, with serological tests. Viral deoxyribonucleic acid was extracted from the infected tissue and amplified using a diagnostic polymerase chain reaction. Sheep of the Rambouillet breed were found to be most susceptible to infection with morbidity and mortality rates of 26.9% and 8.3%, respectively. Morbidity and mortality rates in the entire flock were 18.4% and 6.3%, respectively. The grazing and migration pattern indicates that the disease was probably introduced to the farm by local sheep.

Animals↗

Comparative leukotoxicities of Pasteurella haemolytica isolates from domestic sheep and free-ranging bighorn sheep (Ovis canadensis).

Twenty-eight isolates of Pasteurella haemolytica from domestic sheep (n = 14 isolates) and bighorn sheep (n = 14 isolates) were evaluated for leucotoxicity against peripheral blood neutrophils of bighorn sheep by adding bacterial culture supernatants to bighorn sheep neutrophils in vitro. Leukotoxic isolates of P. haemolytica, defined as causing > 50% neutrophil death as measured by release of lactate dehydrogenase into culture supernatants, were identified from eight of 14 domestic sheep isolates and from 0 of 14 bighorn sheep isolates. The in vitro assay of isolates of P. haemolytica may provide a valid predictive measure of strain virulence of P. haemolytica, and of potential pneumonic episodes in bighorn sheep populations.

Animals↗

The chromosomal distribution and organization of sheep satellite I and II centromeric DNA using characterized sheep-hamster somatic cell hybrids.

A panel of sheep-hamster somatic cell hybrids containing single sheep chromosomes was used to study the chromosomal distribution and organization of two families of sheep centromeric satellite DNA. This study shows that the centromeres of the sheep metacentric chromosomes 1, 2 and 3 differ in their organization and relative quantities of sheep satellite I DNA. The results, when correlated with the proposed formation of these metacentric chromosomes by ancient Robertsonian translocations, suggest a loss or replacement of satellite I centromeric DNA from the centromeres of these sheep chromosomes. Using Southern blot analysis and fluorescence in situ hybridization, this study shows that the recent centric fusion chromosome t2 (rob 9;10) contains little satellite II DNA. Together these results suggest the possibility of substantial reorganization of sheep centromeric DNA families after Robertsonian translocations.

Animals↗