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Susceptibility of Dall sheep (Ovis dalli dalli) to pneumonia caused by Pasteurella haemolytica.

We evaluated susceptibility of Dall sheep (Ovis dalli dalli) to bacterial pneumonia induced by two strains of Pasteurella haemolytica of domestic sheep origin by evaluating the sensitivity of blood neutrophils of eight Dall sheep to lysis by cytotoxins of P. haemolytica, and by intratracheal inoculation of three Dall sheep, two bighorn sheep (Ovis canadensis), and two domestic sheep with 3.7 x 10(6) or 2.5 x 10(7) colony forming units of P. haemolytica. Neutrophils from the Dall sheep were more sensitive to lysis by cytotoxins from supernatants of a P. haemolytica, biotype A, serotype 2 (A2), of domestic sheep origin, than were neutrophils from six bighorn sheep. This cytotoxic bacterium was the same isolate that was used for intratracheal inoculation of two Dall sheep and two domestic sheep. Inoculation of this cytotoxic P. haemolytica A2 resulted in fatal fibrinopurulent pleuropneumonia in the first Dall sheep within 24 hr of inoculation, and pneumonic lesions in the second Dall sheep before it was euthanized 52 hr after inoculation. This strain of P. haemolytica A2 did not cause respiratory disease when inoculated into two domestic sheep. A noncytotoxic strain of P. haemolytica; biotype T, serotype 3,4,10 of domestic sheep origin did not result in pneumonia in the third Dall sheep or two bighorn sheep. Prior to inoculation, P. haemolytica, biotype T isolates were obtained from all three Dall sheep, but none of these isolates was cytotoxic. At necropsy, cytotoxic P. haemolytica A2 was isolated from lungs and other tissues of the two pneumonic Dall sheep. Based on these results, we conclude that Dall sheep appear to be at least as sensitive as bighorn sheep to pneumonia caused by P. haemolytica A2 of domestic sheep origin. Because in vitro and in vivo results appear closely correlated in this and other studies, we believe with additional evaluation and standardization, neutrophil cytotoxicity tests may serve as a substitute for live animal challenges in future studies of pathogenic P. haemolytica in wild sheep.

Animals↗

[Genetic relationships among seven sheep populations using four microsatellite markers].

The genetic polymorphisms of four microsatellite loci BM143, OarHH35, OarAE101, and BMS2508 were analyzed in 286 sheep of seven sheep populations (Small Tail Han sheep, Hu sheep, Ujumqin sheep, Suffolk sheep, Dorset sheep, Charolais sheep, F1 of Dorset male x Small Tail Han female sheep). The numbers of alleles for BM143, OarHH35, OarAE101, and BMS2508 are 9, 11, 14 and 9 in seven sheep populations, respectively. The polymorphism information content/number of effective alleles/ heterozygosity of BM143, OarHH35, OarAE101 and BMS2508 were 0.7073/3.7231/0.7314, 0.8267/6.4399/0.8447, 0.5743/2.5178/0.6028, 0.6172/3.0712/0.6744 in 286 sheep, respectively. The results revealed the greatest genetic variation at OarHH35 locus and the lowest at OarAE101, the greatest genetic variation in Small Tail Han sheep and the lowest in Hu sheep among seven sheep populations. In the unweighted pair group method with arithmetic mean (UPGMA) dendrograms based on Nei's D(A) distance and Nei's D(S) standard genetic distance, the Chinese native breeds (Small Tail Han sheep, Ujumqin sheep, Hu sheep) were grouped together, then with Charolais sheep. The F1 crossbred sheep, and the two British native sheep (Suffolk sheep, Dorset sheep) also clustered together. Microsatellite genotyping in sheep provided a useful tool for examining the genetic relationships among breeds (populations).

Alleles↗

Experiences with morphine injected into the subarachnoid space in sheep.

The effects of morphine (M), 0.1 mg/kg, administered into the lumbosacral subarachnoid space of sheep used for experimental stifle surgery, were investigated. In a pilot study, preservative-free morphine was administered to three sheep, morphine containing preservatives to two sheep, and saline (S) to one sheep. After recovery from anesthesia, all five sheep administered M displayed rear limb weakness. One sheep, which had received morphine containing preservatives, also licked and chewed incessantly at its flank and hindquarters during recovery. A group of 24 sheep was used to study the effects of morphine containing preservatives, injected intrathecally, on recovery from general anesthesia and hindlimb orthopedic surgery. Eight sheep received M, eight sheep received S, and eight sheep had a needle placed in the subarachnoid space without any injection (N). Times from end of anesthesia to standing varied greatly and did not differ significantly among groups (P = .73), with M sheep averaging 119 minutes; S sheep, 87 minutes; and N sheep, 83 minutes. One sheep administered M licked and chewed at its hindquarters during recovery. Another group of 24 sheep was used to study the effects of morphine containing preservatives, injected intrathecally, on postoperative lameness. Treatments were as described previously. Sheep were videotaped intermittently for 36 hours after surgery, and each sheep was scored as follows: 0 = not lame; 1 = slightly lame; and 2 = very lame. The average lameness scores, which did not differ significantly among groups (P = .21), were: M sheep, 1.07; S sheep, 0.81; and N sheep, 0.68. One sheep administered M displayed extensor spasms of the hindlimbs, and could not stand until several hours after surgery. We conclude that subarachnoid morphine at the dosage used produces no apparent benefit in sheep which have had stifle surgery, and in fact may cause detrimental side effects, such as hindlimb weakness, and pruritus or irritation of the hindquarters.

Analgesics, Opioid↗

[Origin and genetic diversity of Mongolian and Chinese sheep using mitochondrial DNA D-loop sequences].

To determine the origin and gene diversity of the Chinese and Mongolian domestic sheep, a partial fragment of mitochondrial DNA D-loop was sequenced for total number of 314 individuals from nine Chinese sheep populations and 11 Mongolian sheep populations. The results show no difference in nucleotide composition between Chinese and Mongolian sheep mtDNA D-loop sequences. However, more variables were identified in Mongolian sheep (26.85% of the sites) than that in Chinese sheep (24.22%). In China, mtDNA haplotype diversity was the highest in Qinghai Tibetan sheep, followed then by Gansu Tibetan sheep, Gansu Alpine Merino, Qinghai Merino, Gannan Tibetan sheep, Small-tailed Han sheep, Tan sheep, Hu sheep and Minxian Black Fur sheep. In Mongolian sheep, mtDNA haplotype diversity was the highest in Bayad and Baidrag populations and the lowest in the Gobi-Altai population. In general, Mongolian sheep have a richer genetic diversity than the Chinese ones with larger number of haplotypes (86.06% (142/165) versus 78.83% (108/137)), higher haplotype diversity (Hd; 0.976 versus 0.936), higher nucleotide diversity (Pi (pi); 0. 036 versus 0.034) and higher average number of nucleotide differences (k; 23.50 versus 22.48). Phylogenetic analysis of the 217 haplotypes identified in both Mongolian and Chinese sheep supported the same origin of their domestication with three distinct maternal lineages defined as major haplotypes A, B and C, of which haplotype A are the commonest in all Chinese sheep populations and in the majority of Mongolian sheep populations (9/11) with an average frequency of 58.73%, followed by haplotype B present in eight of Chinese population and in all Mongolian sheep populations with an average frequency of 24.68%, and haplotype C present in eight Chinese and in 10 Mongolian sheep populations with an average frequency of 16.59%. Further network analysis of the phylogenetic relationship of the 87 haplotypes identified from 91 sequences retrieved from GenBank together with the 217 haplotypes detected in this study reveals clearly four distinct lineages with the European mouflon (O. musimon) mixed into one of the lineages (haplotype B). There is no evidence of contribution of Argali sheep (O. ammon), O. vignei bochariensis and/or O. ammon nigrimontana to the maternal origin of both Mongolian and Chinese domestic sheep.

Animals↗

[Preliminary studies of microsatellite markers OarAE101 and BM1329 in five sheep breeds].

Two microsatellite markers OarAE101 and BM1329 which were closely linked to the fecundity gene FecB in Booroola sheep were analysed for polymorphisms in Small Tail Han sheep, Hu sheep, Charollais sheep, Ujumqin sheep, Dorset sheep, F1 (Dorset x Small Tail Han sheep). The number of alleles for OarAE101 was 5 or 4 in five sheep breeds. The number of alleles for BM1329 was all 4 in five sheep breeds. The polymorphic information contents for OarAE101/BM1329 in Small Tail Han sheep, Hu sheep, Charollais sheep, Ujumquin sheep, Dorset sheep, F1(Dorset x Small Tail Han sheep) were 0.57/0.54, 0.62/0.67, 0.61/0.59, 0.62/0.66, 0.56/0.67, 0.62/0.68, respectively. Least squares mean of litter size for OarAE101 107 bp/113 bp was significantly higher than those for OarAE101 109 bp/109 bp and 107 bp/111 bp (P < 0.05) in Small Tail Han sheep. Least squares means for litter size were not significantly different among other OarAE101 genotypes. The allele 107 bp of OarAE101 had a significant positive correlation with litter size in Small Tail Han sheep, both alleles 109 bp and 111 bp of OarAE101 had a significant negative correlation with litter size in Small Tail Han sheep. Least squares mean of litter size for BM1329 146 bp/158 bp was significantly higher than those for other BM1329 genotypes (P < 0.05) in Small Tail Han sheep. Least squares means for litter size were not obviously different among other BM1329 genotypes. The allele 146 bp of BM1329 had a significant positive correlation with litter size in Small Tail Han sheep, the allele 148 bp of BM1329 had a significant negative correlation with litter size in Small Tail Han sheep.

Alleles↗

[Estrogen receptor as a candidate gene for prolificacy of small tail Han sheep].

Single nucleotide polymorphism in exon 1 of the estrogen receptor (ESR) gene was detected by PCR-SSCP in both high fecundity sheep breeds (Small Tail Han sheep, Hu sheep and German Mutton Merino sheep) and low fecundity sheep breeds (Dorset sheep,Suffolk sheep). Results indicated that there were three genotypes (AA, AB and BB) in all three high fecundity sheep breeds, but only two genotypes (AA, AB) in both low fecundity breeds. In Hu sheep,German Mutton Merino sheep, Small Tail Han sheep, Suffolk sheep and Dorset sheep,the frequency of allele A was 0.672, 0.786, 0.846, 0.857 and 0.867, respectively, and the frequency of allele B was 0.328, 0.214, 0.154, 0.143, and 0.133, respectively. Sequencing revealed a C-->G mutation at 363 bp of exon 1 of ESR gene in the BB genotype in comparison to the AA genotype. The genotype distribution was significantly different between Small Tail Han sheep and Hu sheep (P<0.01) and between Dorset sheep and Hu sheep (P <0.05). There was no difference in genotype distribution between other sheep breeds. The Small Tail Han sheep ewes with genotypes AB or BB had 0.51 (P < 0.05) and 0.7 (P < 0.05) more lambs than those with genotype AA, respectively. These results showed that the estrogen receptor locus is either a major gene that influences the prolificacy in Small Tail Han sheep or in close linkage with such a gene. In view of our results, marker-assisted selection using ESR is warranted to increase litter size in sheep and will be of considerable economic value to mutton producers.

Alleles↗

Study on polymorphisms of microsatellites DNA of six Chinese indigenous sheep breeds.

The polymorphisms of 17 microsatellites loci of six indigenous sheep breeds, including Mongolian sheep, Ujumuqin sheep, Kazakstan sheep, Aletai sheep and Tibetan sheep, were studied using polypropylene gel electrophoresis in order to investigate their genetic diversity,origin, differentiation and relationships. The results indicated that there was a significant difference in genetic diversity between different loci (P < 0.01). There was no significant difference in PIC, Fis and observed heterozygosity (Obs. Het) among populations (P > 0.05), but a significant difference in gene diversity and expected heterozygosity (Exp. Het) (P < 0.05). Chinese indigenous sheep breeds had similar genetic diversity as those from Europe,but with higher inbreeding coefficient. It could be inferred from the cluster of individuals and populations that Chinese indigenous sheep breeds might originate from two ancestors. The cluster of populations showed that Mongolian sheep and Ujumuqin sheep had close relationship and did not differentiate obviously. Mongolian sheep and Tibetan sheep had far relationship and differentiated significantly. Aletai sheep differentiated from Kazakstan sheep but not significantly. Tan sheep,Aletai sheep and Tibetan sheep also had close relationships. The Fst of Chinese indigenous sheep breeds was close to some Spanish sheep breeds,but much smaller than that of other European sheep breeds. More loci and samples should be studied in the future in order to obtain more accurate results.

Animals↗

European Union's rapid TSE testing in adult cattle and sheep: implementation and results in 2001 and 2002.

After the discovery of variant Creutzfeldt-Jakob disease (vCJD), scientific advances quickly led to post-mortem tests to identify late-stage bovine spongiform encephalopathy (BSE) disease. These were first used in Switzerland in 1999 for active BSE surveillance of a) fallen and emergency-slaughter bovines (risk stock) and b) 5% sample of routinely slaughtered cattle over 30 months of age. In 1999 and 2000, Switzerland's estimated 103 BSE positives per 1000000 adult cattle put it in the same BSE risk classification as UK and Portugal. In July 2000, the European Union's Scientific Steering Committee published its methodology (and first vetted results) for geographical BSE risk (GBR) assessment in cattle. Member states with no BSE cases found themselves, on rational assessment, classified as GBR III (BSE likely but not confirmed, or confirmed at a lower level). Because of Europe's thus highly assessed BSE risks, active BSE surveillance of adult cattle in all member states began in January 2001 using one of three validated post-mortem tests. Implementation was variable across member states in January to March 2001 but, where operational, active surveillance was typically achieved for around 13300 routinely slaughtered and 1000 risk stock per month per 1000000 adult cattle; BSE positive rates were 60 and 600 per 1000000 routinely slaughtered and risk cattle, respectively. By the second half of 2001, active BSE surveillance was operating reasonably in most member states, although anomalies persisted. Performance and results for July to December 2001 and for January to June 2002 are considered in detail. The BSE positive rate decreased substantially in UK, Portugal and Ireland between semesters, whereas Spain's rates increased for both routinely slaughtered and risk bovines. Based on 1450000 routinely slaughtered and 135000 risk stock as standard, France could have expected 153 BSE positives in July to December 2001 (109 in January to June 2002); Italy 154 (67); and Germany only 39 (48). When sample-based surveillance data were scaled up and combined with clinical BSE cases, Great Britain's BSE positives were estimated at around 400 per 1000000 adult cattle in 2002 compared with over 1000 per 1000000 adult cattle in 2000. Age distributions for cattle subject to active BSE surveillance have been underexploited. The major transmissible spongiform encephalopathy (TSE) which affects sheep and goats is scrapie. Passive surveillance of scrapie is associated with substantial under-reporting. Susceptibility to scrapie depends strongly on sheep genotype; but resistance to scrapie does not necessarily confer resistance of sheep to BSE. Because of uncertainty about the true prevalence of scrapie-infected adult sheep and concern that BSE in sheep may be missed, the European Union pre-empted its planned evaluation of rapid post-mortem TSE tests in sheep by requiring the rapid TSE testing of small ruminants from April 2002 with one of the three cattle-validated tests. Basic requirements for active TSE surveillance in sheep were: random sample of 6000 fallen sheep and of 60000 routinely slaughtered adult native sheep to be tested per member state by end March 2003. Lower surveillance targets were set for countries with under 1000000 adult sheep. Adequately to map scrapie-susceptible genotypes and identify resistant genotypes, a random sample of 500 routinely slaughtered native adult sheep was to be genotyped, together with each TSE rapid test positive adult sheep and two sets of three suitably sampled controls. By the end of August 2002, when 41% of the initial surveillance time had elapsed, only 20% of the European joint target for routinely slaughtered adult sheep had been completed, but that for fallen sheep was exceeded. Except in Ireland, the upper 95% confidence bound on TSE prevalence exceeded 500 per 1000000 routinely slaughtered adult sheep in reporting-compliant countries with more than 1000000 adult sheep. The UK, Greece, Italy and France were likely to approach the goal of 100 TSE rapid test positives on completion of their assigned first-year surveillance target for sheep. Results from the recommended genotyping of TSE positive adult sheep and controls for use in inferring differential TSE-positive susceptibility by genotype are awaited. Only by genotyping 5000-50000 TSE-positive adult sheep, a massive undertaking even on the European scale, will it become clear whether scrapie resistance is relative rather than absolute. This paper details Europe's quantitative evolution in TSE surveillance.

Animals↗