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Evaluation of a commercially available vaccine against Corynebacterium pseudotuberculosis for use in sheep.

OBJECTIVE: To evaluate the efficacy of a commercially available bacterin-toxoid vaccine for preventing Corynebacterium pseudotuberculosis-induced abscesses in sheep. DESIGN: Prospective randomized controlled trial. ANIMALS: 31 mixed-breed sheep seronegative for C pseudotuberculosis. PROCEDURE: Sheep were randomly assigned to vaccinate (n = 20) or nonvaccinate (11; control) groups. Sheep in the vaccinate group received 2 doses of serial A or serial B bacterin-toxoid vaccine at 4-week intervals. Serologic testing was conducted after vaccination to document an antibody response to vaccination. All sheep were challenge inoculated with virulent C pseudotuberculosis organisms 32 weeks after the second vaccination. Twenty weeks after challenge inoculation, all sheep were examined for external and internal abscesses secondary to C pseudotuberculosis infection. RESULTS: Vaccinated sheep developed an antibody response to both components of the vaccine, as measured by use of ELISA tests. After challenge inoculation, vaccinated sheep had significantly less external, internal, and total abscesses than control sheep. CLINICAL IMPLICATIONS: Vaccination of sheep with a commercially available bacterin-toxoid against C pseudotuberculosis could substantially decrease the prevalence and number of abscesses that form secondary to C pseudotuberculosis infection.

Abscess↗

The influence of chemical form of zinc on the effects of toxic intraruminal doses of zinc to sheep.

The EDTA, sulphate and oxide compounds of zinc were administered to sheep as a single intraruminal dose (480, 240 or 120 mg Zn per kg body weight) or as thrice-weekly doses (240 mg Zn per kg body weight per dose) for 4 weeks. In the single-dose experiment, serum zinc concentrations rose most rapidly, were highest and fell most rapidly in those animals receiving zinc EDTA. In the sheep receiving the sulphate, serum zinc concentrations were high and stayed high for 2 or 3 days. Zinc oxide caused slight, but prolonged, elevation of serum zinc. High concentrations of zinc were present in urine after administration of zinc EDTA. In the multiple-dose experiments, the sheep dosed with zinc sulphate showed progressively higher elevations of serum zinc (first dose, 5-10 micrograms Zn ml-1; day 13, 30-60 micrograms Zn ml-1) and six of the seven sheep so dosed died before the final dose. At post mortem examination, many of these had lesions of the upper gastrointestinal tract and showed evidence of haemolytic crises. Sheep dosed with zinc oxide also had progressively higher peak serum zinc concentrations. In contrast, the peak serum concentrations for the zinc EDTA-dosed sheep declined as the experiment progressed. No deaths occurred in the sheep dosed with zinc oxide or EDTA. Major pancreatic injury occurred in sheep dosed with the sulphate or oxide, but was only mild in the sheep dosed with the zinc EDTA. Diarrhoea was mild and transitory in the EDTA-dosed sheep, but more severe and persistent in those dosed with the sulphate.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Comparison of arachidonate metabolism by alveolar macrophages from bighorn and domestic sheep.

We have defined the metabolites of arachidonic acid (AA) secreted by alveolar macrophages (AMs) of bighorn sheep and domestic sheep in response to three agents: calcium ionophore A23187, phorbol myristate acetate (PMA), and opsonized zymosan. Cells were labeled with [3H]AA prior to stimulation and 11 tritiated metabolites, including prostaglandins (PGs), thromboxanes (TXs), leukotrienes (LTs), and hydroxyeicosatetraenoic acids (HETEs), were detected and quantitated by high-performance liquid chromotography and radiometry. Zymosan stimulation resulted in the release of significantly elevated quantities (P less than 0.05), of LTB4, [5(S), 12(R)-dihydroxy-6,14-cis-8,10-trans-eicosatetraenoic acid], 5-HETE, [5(S)-hydroxyeicosatetraenoic acid], and the nonenzymatic isomers of LTB4, [LTB I, 5(S),12(R)-6-trans-LTB4] and LTB II, [5(S), 12(S)-6-trans-LTB4], from domestic sheep AM when compared to bighorn sheep AM. Phorbol myristate acetate (PMA) stimulation released significantly elevated quantities (P less than 0.04), of TXB2, (thromboxane B2), HHT, [12(S)-12-hydroxy-5,8,10-heptadecaenoic acid], LTB I, LTB II, and 15-HETE, [15(S)-hydroxyeicosatetraenoic acid] from domestic sheep AMs when compared to bighorn sheep AMs. However, after A23187 challenge, only 15-HETE was significantly elevated (P less than 0.04) in domestic sheep AMs when compared to bighorn sheep AMs. These clear differences in AA metabolism of AMs obtained from bighorn and domestic sheep in response to three different agonists suggest not only different control mechanisms for lung metabolism of AA in the two species, but also suggest that differences in the metabolites released may lead to quite different regulation of lung defense mechanisms in the two sheep species.

Animals↗

Six-month daily treatment of sheep with neurotoxic organophosphorus compounds.

The delayed neurotoxic effects of tri-o-cresyl-phosphate (TOCP), O-methyl-O-(4-bromo-2,5-dichlorophenyl) phenylphosphonothioate (leptophos), and O-ethyl O-(4-nitrophenyl) phenylphosphonothioate (EPN) at 5, 5, and 1 mg/kg/day, respectively, on male sheep were studied during 6 months of daily oral treatment under field conditions. A vehicle-control group of sheep given corn oil (0.1 ml/kg/day) only was used for comparison. All sheep were killed 24 h after the 180th daily treatment. Blood, brain, spinal cord, and sciatic nerve tissues were taken for histological and/or biochemical examinations. The results indicated that leptophos induced severe ataxia and paralysis in sheep following about 4 months of treatment. TOCP produced either mild ataxia or lameness in two of four sheep during the last week of experiment. On the other hand, none of the EPN-treated sheep showed clinical signs of neurotoxicity during the course of the experiment at the dosage tested. These clinical results were supported by histological findings and also by biochemical results with neurotoxic esterase (NTE) measurements. In the case of leptophos-treated sheep, numerous prominent degenerative lesions of axons were observed in spinal cords and brains. Similar but somewhat less numerous lesions were noted in sheep treated with TOCP. No histological changes were observed in similar tissues taken from EPN-treated sheep. The results also indicated that, for chronic exposure to these neurotoxic organophosphorus compounds in sheep, a threshold in excess of 60-70% prolonged inhibition of brain NTE, or 50-60% inhibition of spinal cord NTE must be exceeded to initiate clinical and/or histological neurotoxic effects.

Acetylcholinesterase↗

Analysis of prion protein genotypes in relation to reproduction traits in local and cosmopolitan German sheep breeds.

Due to the genetic determination of susceptibility to scrapie and other forms of transmissible spongiform encephalopathy (TSE) in sheep breeding to the less susceptible prion protein (PrP) genotype ARR/ARR was advanced within EU. In 4961 ewes of nine German sheep breeds (Coburg Fox sheep, Gray Horned Heath sheep, Merinoland sheep, Rhoen sheep, German Blackheaded Mutton sheep, Shropshire, Suffolk, Texel and White East Friesian Milk sheep) representing local and cosmopolitan breeds the reproductive traits number of lambs born, dead (including abortion at the end of pregnancy, stillbirth and death during the first 56 days post natum), weaned and rearing rate at each lambing were recorded and in 1641 of these ewes the PrP genotype was determined. A linear model was used to evaluate associations between PrP genotype and reproduction traits including the effects of PrP genotype (four classes: ewes with two, one and no copy of the ARR allele and with unknown PrP genotype), breed, interaction of PrP genotype and breed, number of lambing, lambing season and stock. Significant associations were only observed between the PrP genotype and the number of dead lambs at each lambing in Shropshire and Merinoland sheep and the rearing rate at each lambing in Shropshire. These significant associations were mainly caused by differences between animals with unknown PrP genotype and animals of the other PrP classes. In conclusion, breeding for TSE resistant sheep will not lead to a reduction in economically important reproduction traits.

Alleles↗

VIP innervation: sharp contrast in fetal sheep and baboon adrenal glands suggests differences in developmental regulation.

Immunocytochemical technique and light microscopy were used to ascertain the relationship between vasoactive intestinal polypeptide (VIP) and tyrosine hydroxylase in fetal sheep and fetal baboon adrenal cortices and medullae at 85% of gestation. VIP immunostaining was extremely robust in fetal sheep adrenal cortical neurofibers and cells while weak in fibers and nonexistent in cells of fetal baboon. Also, tyrosine hydroxylase-immunopositive cells, present throughout the adrenal cortices of both fetal sheep and baboons, were heavily innervated by VIP-immunoreactive neurofibers in fetal sheep, but not in fetal baboons. Adrenal cortical VIP-immunopositive fibers occurred in greater (P<0.05) frequency in fetal sheep than in fetal baboons (14.82+/-3.10 vs. 0.84+/-0.26 fibers/field), were larger in diameter (2.93+/-0.34 vs. 0.93+/-0.07 microm) and ran for longer distances in the plane of section (127.85+/-5.16 vs. 74.53+/-4.93 microm). VIP immunogenicity in cells (ganglion and chromaffin) and fibers was robust in fetal adrenal medulla of sheep while nonexistent in baboons. VIP fibers in fetal sheep medulla were smaller in diameter compared to fetal sheep cortex (1.22+/-0.13 vs. 2.93+/-0.34 microm, P<0.05), but not compared to extrinsic nerve fibers (1.30+/-0.09 microm). We hypothesize that in fetal sheep of this age, medullary neurofibers derive primarily from extrinsic sources while cortical fibers arise from cortical ganglion cells. We conclude that at 85% of gestation the potential for VIP neural control of paracrine (e.g., glucocorticoid/catecholamine) interactions in both adrenal cortex and medulla is much greater in fetal sheep compared to fetal baboons.

Adrenal Cortex↗

Evaluation of bluetongue virus diagnostic tests in free-ranging bighorn sheep.

Five bluetongue virus (BTV) diagnostic tests were evaluated for use in free-ranging bighorn sheep. We sampled one bighorn sheep population four times between 1989 and 1995. The tests evaluated included virus isolation (VI), polymerase-chain reaction (PCR), serum neutralization (SN), agar-gel immunodiffusion (AGID), and competitive enzyme-linked immunosorbent assay (c-ELISA). The c-ELISA, AGID and SN tests had high levels of agreement in determining serogroup exposure in bighorn sheep. We used maximum-likelihood algorithms to estimate the parameters of each diagnostic test used. Although the c-ELISA and AGID had high sensitivity and specificity, the SN had perfect specificity but lower apparent sensitivity. Due to the potential of cross-reactions among multiple serotypes, results of the SN must be interpreted with caution when assessing serotype exposure in an area where multiple serotypes are endemic. The PCR assay delineated convalescent antibody titers from more-recent infections, and consequently, was pivotal in distinguishing a different exposure pattern between the bighorn sheep and cattle in an adjacent herd. Based on an increasing seroprevalence (50% to 100%), BTV circulated through this bighorn sheep population between 1989 and 1993. This increase in seroprevalence coincided with a bighorn die-off due to BTV infection in June, 1991. An adjacent cattle herd was sampled in 1995 for comparison. The bighorn sheep and adjacent cattle had different patterns of exposure to BTV between 1994 and 1995. There was no evidence that BTV circulated through the bighorn sheep population from 1994 to 1995. In 1995, seroprevalence to BTV decreased to 72%, none of yearling bighorn was seropositive, and all of the 39 bighorn sheep were PCR-negative. In contrast, all adult cattle were seropositive to BTV by c-ELISA and SN, and 4 of the calves were seropositive; 11 of the 24 cattle were PCR-positive, including all five calves. Overall, the pattern of temporal herd immunity in the bighorn sheep appeared to follow a classic epidemic curve, with the appearance and subsequent disappearance of herd immunity coinciding with the 1991 die-off in this population. As low levels of herd immunity and high proportions of susceptible animals are key factors in the development of epidemics, this population of bighorn sheep may be at increased risk for a BTV epidemic in the future.

Animals↗

Rates of entry and oxidation of acetate, glucose, D(-)-beta-hydroxybutyrate, palmitate, oleate and stearate, and rates of production and oxidation of propionate and butyrate in fed and starved sheep.

1. Rates of entry and oxidation of a range of metabolites have been measured in tracheostomized sheep (diet, 800g. of lucerne chaff and 100g. of maize/day) by combining isotope-dilution techniques with the continuous measurement of total respiratory gas exchange, and (14)CO(2) production during the intravenous or intraruminal infusion of (14)C-labelled substrates. 2. Mean entry rates in fed and starved (24hr.) sheep respectively, expressed as mg./min./kg. body wt.(0.75), were: glucose, 5.0 (range 4.8-5.1, 2 observations) and 3.8 (3.2-4.2, 4); acetate, 10.8 (9.1-13.5, 4) and 5.8 (1); d(-)-beta-hydroxybutyrate, 1.4 (1) and 1.5 (0.8-2.4, 4); palmitate, oleate and stearate (starved sheep only) 1.0 (0.6-1.9, 7), 0.9 (0.2-1.6, 10) and 0.9 (0.5-1.1, 11) respectively. 3. Production rates of propionate and butyrate in continuously feeding sheep were 6.4 (4.7-8.3, 4) and 4.3 (3.4-6.1, 4) mg./min./kg.(0.75) respectively, and in starved (24hr.) sheep were 2.5 (2.2-2.9, 2) and 1.0 (0.8-1.2, 2) mg./min./kg.(0.75) respectively. 4. Calculated terminal values for the specific radioactivity of respiratory (14)CO(2) during measurements of entry rates and production rates were used to calculate the contributions of individual substrates to overall oxidative metabolism. Mean values for fed and starved sheep respectively were: glucose, 9.1 (8.6-9.6, 2) and 11.2 (5.9-15.1, 4)%; acetate, 31.6 (26.8-38.1, 4) and 22.1 (1)%; d(-)-beta-hydroxybutyrate, 10.4 (1) and 4.8 (1.9-7.7, 4)%; propionate, 23.0 (13.8-29.9, 4) and 7.1 (6.8-7.4, 2)%; butyrate, 16.5 (13.7-20.5, 4) and 5.3 (5.2-5.3, 2)%; palmitate, oleate and stearate (starved sheep only), 4.7 (2.0-7.7, 7), 4.0 (1.2-6.6, 10) and 4.4 (3.8-5.8, 9)% respectively. The sum of these values for individual substrates in fed and starved sheep, excluding that of beta-hydroxybutyrate and after correction of the glucose value for the known interrelations of this substrate with propionate, accounted for 76% and 58% respectively of total production of carbon dioxide. 5. Calculations based on the proportion of substrate entry directly oxidized indicated that the substrates studied accounted for 63% (fed sheep) and 43% (starved sheep) of total energy expenditure measured by oxygen uptake. The contribution of beta-hydroxybutyrate was excluded, and corrections were made for glucose-propionate interrelations, and for the different rates of oxidation of the methyl and carboxyl fragments of acetate. 6. The present results have been combined with those obtained earlier in this Laboratory to examine the relationships between rates of substrate entry and oxidation, and concentrations of substrate in blood. Rates of entry of acetate, glucose, d(-)-beta-hydroxybutyrate, palmitate and oleate (but not stearate) were well correlated with concentration in blood, and substrate contribution to production of carbon dioxide showed a similar correlation to blood concentration, except with glucose. 7. It was concluded that the general technique is of potential value in providing valid quantitative parameters of animal metabolism.

Acetates↗

Allergy to goat and sheep cheese with good tolerance to cow cheese.

BACKGROUND: We report on a patient who experienced allergic reactions after eating goat cheese and after touching goat and sheep cheese, but not after consuming cow's milk dairy products. OBJECTIVE: To assess the allergenicity and IgE-binding capacity of the caseins from the three different species. METHODS: Skin prick tests were carried out using whole milk and caseins from three different species (goat, sheep and cow), and whey fractions of cow's milk. Total serum IgE and specific IgE to cow's milk proteins were measured by CAP system and specific IgE against caseins and whole milk were determined by ELISA technique. To evaluate allergenic cross-reactivity, inhibition of the IgE ELISA activity to goat's milk and goat casein was tested for the three caseins. SDS-PAGE and immunoblotting was used to determine IgE binding bands in caseins. RESULTS: Skin tests were positive to sheep and goat's milk, sheep and goat casein, as well as to sheep and goat cheese. Total serum IgE was 66 kU/L and IgE determinations by CAP were negative. IgE ELISA against the caseins from goat and sheep was strongly positive, whereas it was negative to cow casein. ELISA inhibition assays revealed a high degree of cross-reactivity between goat casein and sheep casein. Immunoblotting showed three IgE-binding bands in goat casein at 31, 27 and 22 kDa, which may correspond to alpha-, beta- and gamma-caseins. A band at about 31 kDa was observed in sheep casein and another band at 34 kDa was recognized in cow casein. CONCLUSION: This patient developed allergy to goat and sheep cheese with good tolerance to cow's milk. We identified goat casein as the main allergen causing sensitization in this patient as demonstrated by in vivo and in vitro tests. A high degree of cross-reactivity between goat and sheep casein was observed.

Allergens↗

Sequences and diversity of 17 new Ovar-DRB1 alleles from three breeds of sheep.

To investigate the genetic diversity of the sheep MHC (Ovar) class II DRB1 locus, we amplified exon 2 of Ovar-DRB1 alleles by polymerase chain reaction (PCR) and determined the nucleotide sequences of both resultant strands after cloning. In our study of a total of 97 sheep of three breeds, namely, Suffolk, Cheviot and Corriedale, we identified 18 previously published alleles and 17 new alleles. These alleles were 83.4 to 94.1% identical at the nucleotide level and 71.4 to 90.9% identical at the amino acid level to Ovar-DRB1*0101. We identified six new alleles in Cheviot sheep and 11 new alleles in Suffolk sheep. Furthermore, we identified 15, 6 and 1 allele in Suffolk, Cheviot and Corriedale sheep, respectively, that have only been found in these breeds to date. Analysis of the frequencies of the various Ovar-DRB1 alleles in each breed indicated that Ovar-DRB1*0702 was the most frequent allele in Suffolk sheep (23.9%), Ovar-DRB1*0203 was the most frequent allele in Cheviot sheep (27.5%) and Ovar-DRB1*0201 was the most frequent allele in Corriedale sheep (25.0%). A comparative analysis of the positions of polymorphic residues in the first extracellular domain of the DRB genes of sheep, humans and mice revealed an extraordinary similarity amongst the positions of polymorphic residues that are associated with the antigen recognition site (ARS). Moreover, the extent of polymorphism seems to be similar in sheep, humans and mice.

Amino Acid Sequence↗

Vacuolar lesion profile in sheep scrapie: factors influencing its variation and relationship to disease-specific PrP accumulation.

Detailed neuropathological examination for vacuolar lesions was performed on the brains of 42 sheep with clinical signs compatible with scrapie. The sheep were grouped according to their breed (Poll-Dorset, Cheviot, Welsh Mountain, Shetland and Suffolk), their PrP genotype at codons 136, 154 and 171 (VRQ/VRQ, VRQ/ARQ, VRQ/ARR and ARQ/ARQ) and the type of infection (experimental infection with SSBP/1, or natural disease). Twenty-two neuroanatomical sites from seven brain regions were examined for vacuolation in the neuropil and five sites at the level of the obex were examined for intraneuronal vacuolation. In 36 sheep, immunohistochemical examination for disease-specific PrP (PrP(d)) accumulation had also been performed in the same brain regions in an earlier study. The magnitude of total neuropil vacuolation was highest in the naturally affected ARQ/ARQ Suffolk sheep and lowest in the experimentally infected VRQ/VRQ Cheviot sheep and VRQ/ARR Poll-Dorset sheep. The severity of neuropil vacuolation at nine of the 22 neuroanatomical sites examined was used to generate a vacuolar lesion profile, which showed variations between the different sheep groups. These variations could be attributed to both PrP genotype and sheep breed and also possibly to scrapie agent; there was, however, considerable individual variation in lesion profile within sheep groups. All groups showed a similar ratio of neuropil vacuolation to neuronal vacuolation at the level of the obex. Although a positive correlation between neuropil vacuolation and PrP(d) deposition was generally observed, it was low except for the astrocyte-associated pattern of PrP(d) accumulation. The study suggests that vacuolar lesion profiles in sheep are affected by several factors and, by comparison with lesion profiles in mice, are of no more than limited value for discriminating between scrapie strains.

Animals↗

Studies of the large intestine of sheep. 1. Fermentation and absorption in sections of the large intestine.

1. Fermentation and absorption of constituents of digesta in segments of the large intestine of sheep given different diets were studied by analysis of gut contents obtained at slaughter after a period during which the sheep had been administered a non-absorbable gut marker. 2. In sheep given chopped, dried lucerne (Medicago sativa) there was net absorption of water throughout the large intestine with concomitant increases in the proportion of dry matter (DM) and organic matter (OM). There was net disappearance of 62 g OM, 1.66 g non-urea non-ammonia-nitrogen (NU-NAN) and 0.6 g (urea + NH3(-N in the caecum and proximal colon. There was no significant change in OM and NU-NAN flow through the remainder of the large intestine but there was a net disappearance of 0.3 g NH3-N. There was also net appearance of volatile fatty acids (VFA) in the caecum, most of which was apparently absorbed before the rectum. 3. Metabolism in the caecum was also studied in sheep grazing fresh pasture or consuming one of three sugar cane-bagasse-based diets, or barley pellets. In the lucerne- and pasture-fed sheep there was a net disappearance of approximately 0.5 g NH3-N/d from the caecum, while in sheep fed on bagasse plus urea, 1.4 g NH3-N/d was apparently absorbed from this region. The addition of fish meal to this latter diet resulted in apparent disappearance of 5.3 g NH3-N/d from the caecum and proximal colon. 4. There was apparent loss of NU-NAN from the caecum of sheep on all diets except the barley diet. With the latter diet there was a net gain of 1 g NU-NAN/d which was associated with relatively high VFA concentration and production; taken together these results indicate that microbial fermentation in the caecum was more extensive in the sheep fed on the barley diet than in those fed on the other diets. 5. The proportions of individual VFA in digesta from the rumen and caecum of lucerne-fed and pasture-fed sheep and in digesta from the caecum of sheep given the bagasse-based or barley diets are also reported and discussed. 6. In general the results indicate that the caecum and to a lesser extent the proximal colon were the major regions of fermentation and absorption of the components of the digesta in the large intestine.

Ammonia↗

Effect of food intake on energy and protein metabolism in the skin of Romney sheep.

Sheep fed on either a low (500 g lucern (Medicago sativa) chaff/d; L) or high (1100 g lucerne chaff/d; H) intake had measurements made, using arterio-venous techniques, of blood flow and energy metabolite and cysteine utilization in the skin. Sheep on the H intake had significantly increased skin blood flow (P = 0.014) and oxygen uptake (P = 0.05). Although the H sheep had higher skin blood flow they showed no difference in skin uptake of either glucose or acetate compared with the L sheep, but the H sheep had a significantly lower output of lactate (P = 0.014). Animals in each group had either [14C]glucose or [14C]acetate infused into the skin which showed that acetate was the predominant precursor of skin sterol and fatty acid synthesis in the H sheep while L sheep skin used both glucose and acetate. The H sheep showed an increase in the net uptake of cysteine by the skin (P = 0.053), and in the uptake of cysteine for protein synthesis (P = 0.078), relative to the L sheep and this increase was of a comparable magnitude to the increase in blood flow to the skin. Although blood flow, protein synthesis and energy supply increased in the skin of the H sheep by 200-300%, wool production would only have increased by 10-20%, suggesting that nutrient flux changes are not the sole level of regulation of wool production.

Acetates↗

Association between PCR-SSCP of growth differentiation factor 9 gene and high prolificacy in Small Tail Han sheep.

Small Tail Han sheep that has significant characteristics of high prolificacy and nonseasonal ovulatory activity is an excellent local sheep breed in P.R. China. The lambing percentage averaged 260% in Small Tail Han sheep. Growth differentiation factor 9 (GDF9) gene, which was essential for growth and differentiation of early ovarian follicles, was considered as a possible candidate gene for litter size in Small Tail Han sheep. The genetic polymorphism of a part of the GDF9 gene was detected in 130 ewes of Small Tail Han sheep by PCR-SSCP. The results indicated that there were two genotypes (AA and AB) detected by two primer pairs. In both exon 1 and exon 2 of the GDF9 gene in Small Tail Han sheep, frequencies of AA genotype were 0.846 and 0.908, frequencies of AB genotype were 0.154 and 0.092, frequencies of A allele were 0.923 and 0.954, and frequencies of B allele were 0.077 and 0.046, respectively. The results of chi2 fitness test indicated that both exon 1 and exon 2 of the GDF9 gene were in Hardy-Weinberg equilibrium (p > 0.05) in Small Tail Han sheep. Least squares means of litter size in the first and the second parity for genotype AA were 0.30 (p <0.05) and 0.77 (p <0.0001) more than those for genotype AB detected in exon 1 of the GDF9 gene in Small Tail Han sheep, respectively. Fragments detected in exon 2 of the GDF9 gene had no significant effect (p > 0.05) on litter size in both the first and the second parity in Small Tail Han sheep. Litter size in sheep is lowly heritable, expressed only in females, and manifested relatively late in life. Access to genetic markers would thus be advantageous in selection programs.

Alleles↗

Gonial angle changes after bilateral unidirectional mandibular distraction in sheep.

It was the aim of this study to investigate radiologically the distraction site in the gonial angle area after three different protocols of unidirectional mandibular distraction in sheep. Bilateral mandibular distraction was performed in nine sheep, and one other sheep served as a sham-operated control. In the nine experimental sheep, three different distraction protocols were used. In three sheep, an oblique corticotomy line was applied and the distractors were positioned perpendicular to the corticotomy line. In three other sheep, a horizontal corticotomy line was applied and the distractors were also positioned perpendicular to this corticotomy line. In the remaining three sheep, again, a horizontal corticotomy was performed, but with an angular positioning of the distractors relative to this corticotomy line. Distraction of 20 mm was performed over 20 days at a rate of 1 mm per day. After 3 weeks of retention the sheep were sacrificed, the mandibles were dissected and hemisectioned, and standardized hemimandibular contact radiographs were made of the specimens. The distracted gaps were traced from the radiograph and the gonial angle was measured. The displacements of the distracted parts were measured, the gonial angles were determined, and the values were compared with four additional control non-distracted sheep mandibles. The results show that there was a very large variability in the distracted gaps as well as in the gonial angle changes after distraction, and it was not possible to recognize any of the three different distraction protocols radiographically after distraction. Although performed under standardized conditions it is difficult to reproduce intended mandibular corticotomy lines in sheep.

Animals↗

Phylogenetic analysis of snow sheep (Ovis nivicola) and closely related taxa.

Based on mitochondrial cytochrome b gene sequence analysis, the history of true sheep (Ovis) began approximately 3.12 million years ago (MYA). The evolution of Ovis resulted in three generally accepted genetic groups: Argaliforms, Moufloniforms, and Pachyceriforms. The Pachyceriforms of the subgenus Pachyceros comprise the thin-horn sheep Ovis nivicola (snow sheep), Ovis dalli (Dall and Stone sheep), and Ovis canadensis (Rocky Mountain and desert bighorn). North America wild sheep (O. canadensis and O. dalli) evolved separately from Eurasian wild sheep and diverged from each other about 1.41 MYA. Ancestral stock that gave rise to snow sheep, Moufloniforms, and Argaliforms occurred 2.3 MYA, which then gave rise to two different extant lines of snow sheep that diverged from each other about 1.96 MYA. The more recent nivicola line is genetically closer to the North American wild sheep and may represent a close association during the refugium when Alaska and Siberia were connected by the Bering land bridge. The earlier period of evolution of the Pachyceriforms suggests they may have first evolved in Eurasia, the oldest ancestor then giving rise to North American wild sheep, and that a canadensis-like ancestor most likely gave rise to nivicola. Cytogenetic analysis further validates that the standard diploid number for modern nivicola is 52.

Animals↗

Nitric oxide synthase inhibition versus norepinephrine for the treatment of hyperdynamic sepsis in sheep.

OBJECTIVES: To investigate the effects of Nomega-mono-methyl-L-arginine (L-NMMA), an inhibitor of nitric oxide synthesis, on hemodynamics, oxygen transport, and regional blood flow in an ovine model of hyperdynamic sepsis and to compare these effects with the responses to norepinephrine. DESIGN: Prospective, nonrandomized, controlled experimental study with repeated measures. SETTING: Investigational intensive care unit at a university medical center. SUBJECTS: Twenty-five female, healthy, adult sheep of the Merino breed, divided into three groups: nine control sheep; eight sheep treated with L-NMMA; and eight sheep treated with norepinephrine. INTERVENTIONS: All sheep were chronically instrumented. After a 5-day recovery period, a continuous infusion of live Pseudomonas aeruginosa (2.5 x 10(6) colony-forming units/min) was started and maintained for the remainder of the experiment. After 24 hrs of sepsis, eight sheep received L-NMMA (7 mg/kg/hr), eight sheep received norepinephrine, and nine sheep received the vehicle alone (0.9% saline). The norepinephrine dosage was continuously and individually adjusted to achieve the same increase in blood pressure as was observed in a matched sheep of the L-NMMA group. MEASUREMENTS AND MAIN RESULTS: After 24 hrs of sepsis, all sheep developed a hyperdynamic circulatory state with increased cardiac indices and reduced arterial pressures, and systemic vascular resistances. L-NMMA reversed the hyperdynamic circulation, causing an increase in arterial pressure by peripheral vasoconstriction. Norepinephrine led to an increase in blood pressure by augmenting cardiac indices, leaving the systemic vascular resistance unaffected. The norepinephrine dose needed to keep the blood pressure high had to be continuously increased, reflecting the reduced vascular responsiveness to catecholamines during sepsis. Renal blood flow remained unaffected by all treatment forms. Norepinephrine and L-NMMA led to a dramatic increase in urine production. Blocking the nitric oxide synthase with L-NMMA did not interfere with the host's pulmonary ability to clear bacteria, nor did treatment with norepinephrine. CONCLUSIONS: Blocking nitric oxide synthase had a marked vasoconstrictive effect. Both norepinephrine and L-NMMA increased arterial pressure without reducing renal blood flow, leading to an improved renal function.

Animals↗

Selective inhibition of inducible nitric oxide synthase: effects on hemodynamics and regional blood flow in healthy and septic sheep.

OBJECTIVES: To investigate the effects of S-ethylisothiourea (S-EITU) on hemodynamics, oxygen transport, and regional blood flow in healthy and septic sheep. DESIGN: Prospective, randomized, controlled experimental study with repeated measures. SETTING: Investigational intensive care unit at a university medical center. SUBJECTS: Eleven healthy, female adult sheep of the Merino breed, divided into a control group (n = 5) and into a group treated with S-EITU (n = 6). INTERVENTIONS: All sheep were chronically instrumented. After a 5-day recovery period, they were randomly assigned to either control or S-EITU groups. While control sheep received only saline, S-EITU was administered in increasing doses of 1, 3, and 9 mg/kg/hr over 1 hr each (nonseptic phase). After 2 days of recovery, a continuous infusion of live Pseudomonas aeruginosa (2.5 x 106 colony-forming units/min) was started in all sheep and maintained for the remainder of the experiment. After 24 hrs of sepsis, the sheep again received their assigned treatment (septic phase). In both the nonseptic and septic phases, the sheep received colored microspheres through a left atrial catheter to allow analysis of regional blood flows. All animals were autopsied at the end of the experiments, and organ probes were removed for blood flow analyses. MEASUREMENTS AND MAIN RESULTS: The administration of S-EITU caused a dose-dependent vasoconstriction in the nonseptic phase. After 24 hrs of Pseudomonas infusion, all sheep developed a hyperdynamic circulatory state, with increased cardiac indices and reduced arterial pressures and systemic vascular resistances. Oxygen extraction decreased significantly, preventing an increase in oxygen consumption, despite an increased oxygen delivery. The hyperdynamic circulation was dose dependently reversed by S-EITU, causing an increase in arterial pressure by peripheral vasoconstriction. Sheep in the control group showed a continuation of the hyperdynamic circulation. The effects of S-EITU on hemodynamics and regional blood flows were comparable under septic and nonseptic conditions. CONCLUSIONS: With the inducible form of nitric oxide synthase expressed under septic, but not under nonseptic conditions, S-EITU was expected to have vasoconstrictive properties only in the septic phase. It produced a comparable vasoconstriction during the nonseptic phase of the experiment. Thus, either S-EITU does not selectively block the inducible nitric oxide synthase in sheep, or other vasodilators besides nitric oxide play an important role in septic vasodilation.

Animals↗