Bloat in sheep (Ovis aries)

1. Most of the field studies on bloat are conducted with cattle and most of the laboratory experiments seeking to explain the various parameters associated with bloat are done with sheep. 2. Based on grazing behaviour, it would be expected that sheep might bloat more severely than cattle because they selectively choose to eat leaves over stems and chew what they ingest more frequently than cattle. Furthermore, sheep appear to select legumes over grasses because the legumes can be eaten more rapidly. However, because they are selective, sheep eat more slowly than cattle. Despite a higher bloat expectation, bloating in sheep is reported to be less of a problem than in cattle. 3. Although frothing of rumen ingesta was described earlier in cattle as the cause of acute legume bloat, experiments with frothy bloat in sheep preceded those in cattle. 4. Anti-frothing agents were used in sheep before cattle to treat acute legume bloat. 5. Experiments devoted to the study of eructation in ruminants were carried out on sheep, then cattle. 6. Convincing evidence that rumen motility does not cease during acute legume bloat was gathered using sheep. 7. Although the transected tracheal technique for the determination of the volume of eructated gas was developed with cattle, the pathway of eructated gas was confirmed with sheep. 8. All the current evidence accumulated from experiments with sheep supports the hypothesis that death due to legume bloat is caused by acute neural, respiratory, and cardiovascular insult resulting from the effect of the distended rumen on thoracic viscera, diaphragm, intercostal muscles, and the abdominal vena cava. 9. Experiments with sheep and cattle being fed scabrous and nonscabrous diets similar in chemical composition show that sheep are more resistant than cattle to the increase in intrarumen pressure, decline in rumen contraction amplitude, and decrease in rumen contraction frequency caused by nonscabrous diets. 10. The sequence of events in the reticulorumen during primary and secondary contractions previously described following visual and palpation experiments with cattle was confirmed by the use of myoelectrodes implanted in the various sacs of the reticulorumen of sheep. 11. Elevated intrarumen pressure is associated with an increase in the frequency of primary (mixing) and secondary (eructation) contractions (more secondaries than primaries).(ABSTRACT TRUNCATED AT 400 WORDS)     

Differentiation and isolation of hepatic-like cells from human embryonic stem cells

Human embryonic stem cells are pluripotent cells that can serve as a cell source for transplantation medicine, and as a tool to study human embryogenesis. We investigate here the potential of human embryonic stem cells to differentiate into hepatic cells. We have characterized the expression level of liver-enriched genes in undifferentiated and differentiated human embryonic stem cells by DNA microarrays. Our analysis revealed a subset of fetal hepatic enriched genes that are expressed in human embryonic stem cells upon differentiation into embryoid bodies. In order to isolate the hepatic-like cells, we introduced a reporter gene regulated by a hepatocyte-specific promoter into human embryonic stem cells. We isolated clones of human embryonic stem cells that express enhanced green fluorescent protein upon in vitro differentiation. Through immunostaining, we showed that most of these cells express albumin, while some cells still express the earlier expressed protein alpha-fetoprotein. Using fluorescence activated cell sorter, we were able to sort out the fluorescent differentiated cells and expand them for a few more weeks. This is the first report to demonstrate the possibility of purifying differentiated derivatives of human embryonic stem cells and culturing them further. Through confocal microscopy, we detected clusters of hepatic-like cells in 20-day-old embryoid bodies and in teratomas. As observed during embryonic development, we showed that in teratomas, the hepatic-like endodermal cells develop next to cardiac mesodermal cells. In order to examine the secreted factors involved in the induction of hepatic differentiation, human embryonic stem cells were grown in the presence of various growth factors, demonstrating the potential involvement of acidic fibroblast growth factor in the differentiation. In conclusion, given certain growth conditions and genetic manipulation, we can now differentiate and isolate hepatic-like cells from human embryonic stem cells.     

In vitro gametogenesis from embryonic stem cells

Many insights into mammalian germ cell development have been gained through genetic engineering and in vivo studies, but the lack of an in vitro system for deriving germ cells has hindered potential advances in germ cell biology. Recent studies have demonstrated embryonic stem cell differentiation into germ cells and more mature gametes, although significant unanswered questions remain about the functionality of these cells. The derivation of germ cells from embryonic stem cells in vitro provides an invaluable assay both for the genetic dissection of germ cell development and for epigenetic reprogramming, and may one day facilitate nuclear transfer technology and infertility treatments.     

The Complete Mitochondrial DNA Sequence of the Domestic Sheep (Ovis aries) and Comparison with the Other Major Ovine Haplotype

The complete mitochondrial DNA (mtDNA) molecule of the domestic sheep, Ovis aries, was sequenced, together with part of the mtDNA of a specimen representing the other major O. aries haplotype group. The length of the complete ovine mtDNA presented is 16,616 nucleotides (nt). This length is not absolute, however, due to heteroplasmy caused by the occurrence of different numbers of a 75-nt-long tandem repeat in the control region. The sequence data were included in analyses of intraspecific ovine molecular differences, molecular comparisons with bovine mtDNAs, and phylogenetic analyses based on complete mtDNAs. The comparisons with bovine mtDNAs were based on the central domains of the ovine control regions, representing both major ovine haplotype groups, and the corresponding domains of Bos taurus and B. indicus. The comparisons showed that the difference between the bovids was 1.4 times greater than the intraspecific ovine difference. These findings suggest that the strains of wild sheep from which domestic sheep originated were more closely related than were the B. primigenius subspecies which gave rise to B. indicus and B. taurus cattle. Datings based on complete mtDNAs suggest that the bovine and ovine lineages diverged about 30 million years before present. This dating is considerably earlier than that proposed previously. Received: 5 September 1997 / Accepted: 5 May 1998     

Blood biochemistry values of sheep (Ovis aries ligeriensis)

1. Analysis of biochemical parameters were carried out on material pooled from 30 female sheep (Ovis aries ligeriensis). 2. The values determined were for the common metabolites and enzymes utilized for specific studies in general metabolism (urea, glucose, cholesterol, lipids, bilirubin, uric acid, creatinine, alkaline phosphatase, GOT, GPT, LDH, LAP, CGT, CK and amylase). 3. Results of these studies were compared with values from normal human adults. 4. The differences obtained in human and sheep ranges can be explained by the different physiology of the two species. 5. This study gives values for the sheep as an experimental animal in biomedical research.     

Sister chromatid exchange in chromosomes of sheep (Ovis aries)

Blood lymphocyte cultures from 32 Comisana and Laticauda sheep breeds (15 males and 17 females) raised in Southern Italy were studied using sister chromatid exchange (SCE) test. Of the 932 cells studied, the SCE-mean value was 7.20 +/- 2.5 per cell for both breeds. Indeed, the SCE mean values were 7.12 +/- 2.45 and 7.28 +/- 2.55 in Comisana and Laticauda breeds, respectively, and the differences were not significant. No statistical differences were noticed between male and female cells (7.25 +/- 2.39 and 7.16 +/- 2.60, respectively). The SCE frequency distribution did not follow a Poisson distribution. The number of SCE were significantly higher than expected in chromosomes 1, 2 and 3 (p < 0.001) and significantly lower than expected in the X and remaining chromosomes (p < 0.001) on the basis of relative chromosome lengths.     

Physiological and genetic characteristics of black-boned sheep (Ovis aries)

Here we report for the first time the discovery of sheep that have black bones and black muscles. The spectral pattern of pigment extracted from tissues of these black-boned sheep is similar to that of black-boned Chinese silky fowl. Additionally, black-boned sheep have significantly higher plasma colour, tyrosinase activity and kidney function than normal sheep. Synonymous nucleotide substitutions in the tyrosinase (TYR) and melanocortin 1 receptor (MC1R) genes were detected in black-boned sheep when compared with the corresponding sequences in normal sheep. In addition, a missense mutation (215T>C) in exon 2 of tyrosinase-related protein 1 (TYRP1) was detected in black-boned sheep, and this resulted in a putative valine-to-alanine substitution at codon 68 (Val68Ala).     

Primary structure of the ram (Ovis aries) protamine

The amino acid sequence of the protamine isolated from mature sperm nuclei of the ram (Ovis aries) has been established from automated sequence analysis of the S-carboxymethylated protamine. Ram and bull protamines differ only by two point changes and the deletion in bull protamine of the tripeptide Cys39-Arg-Arg41. In mammalian protamines the central region (residues 13-36) consisting mainly of arginine clusters appears to be conserved whereas the N-terminal and C-terminal regions are more variable.     

Effects of insulin and glucose on the production and utilization of glucose in sheep (Ovis aries)

1. The rates of appearance (RA) and metabolic clearance rates (MCR) of glucose were determined during the infusion of insulin plus glucose in sheep. 2. During euglycaemia 50% inhibition of RA of endogenous glucose occurred at insulin concentrations of 60 microU/ml in arterial plasma. 3. During euglycaemia the MCR of glucose doubled with an increment of about 100 microU/ml of insulin. 4. Hypoglycaemia was associated with a reduction in insulin's suppression of RA of endogenous glucose and hyperglycaemia significantly reduced the increase in MCR due to insulin.     

A novel maternal lineage revealed in sheep (Ovis aries)

It is generally believed that domestic sheep have two maternal lineages (haplotypes A and B), based on mitochondrial DNA analysis. In the present study, we provide evidence that a novel maternal lineage (haplotype C) is exhibited in Chinese native sheep. To verify this finding, 231 samples were collected from six Chinese local breeds, which cover the vast geographical region of sheep inhabitation in China. For comparison, 50 samples were collected from two Western breeds collected in China. Mitochondrial DNA was screened by PCR single-strand conformational polymorphism (SSCP), leading to the identification of novel band patterns in ND2 and ND4 genes in the Chinese breeds. Interestingly, mutations at the two loci were in strong linkage disequilibrium. Direct sequencing of the DNA fragments revealed a non-synonymous substitution in ND2. Furthermore, two synonymous mutations were identified by comparisons of the novel type (haplotype C) and the established types (haplotypes A and B). The entire mitochondrial control region for 55 samples was then sequenced to construct a phylogenetic tree and median joining network. Both the tree and network demonstrated a topology of three groups, which is in consistent with the SSCP analysis. Unlike Western breeds, Chinese breeds are composed mainly of haplotypes A and B, but with a small fraction of haplotype C. According to Fu's test and mismatch distribution, haplotype C has not been subject to a recent population expansion. Based on these results, we propose a novel origin for Chinese sheep.     

Magnitude of ouabain-sensitive respiration of lamb hepatocytes (Ovis aries)

In lamb hepatocyte preparations with viabilities greater than 90%, ouabain-sensitive respiration accounted for approximately 50% of the total cellular O2 consumption. Lamb hepatocyte preparations with viability of less than 50% exhibited lower (P less than 0.05) total and ouabain-sensitive respiration. The decrease in ouabain-sensitive respiration in these preparations entirely accounted for the drop in total respiration.     

Resolving ambiguities in the karyotype of domestic sheep (Ovis aries)

Internally consistent G-, Q- and R-banded karyotypes and idiograms for sheep chromosomes at the 422-band level of resolution are presented. These were derived by sequential Q- to G-staining, and sequential Q- to R-staining of prometaphase spreads prepared from sheep with normal and Robertsonian chromosomes. The fused chromosomes served as stable morphological markers. To minimise confusion due to chromosomal nomenclature, we have listed chromosome-specific (reference) molecular markers that have been mapped byin situ hybridization to sheep chromosomes. The use of molecular markers in conjunction with the sequential Q- to G- and sequential Q- to R-banded karyotypes and iodiograms provided here will elimiate ambiguities in identifying and numbering sheep chromosomes and will facilitate their comparison with cattle chromosomes. Edited by: J.B. Rattner     

Mitochondrial DNA variation of domestic sheep (Ovis aries) in Kenya

The history of domestic sheep (Ovis aries) in Africa remains largely unknown. After being first introduced from the Near East, sheep gradually spread through the African continent with pastoral societies. The eastern part of Africa was important either for the first diffusion of sheep southward or for putative secondary introductions from the Arabian Peninsula or southern Asia. We analysed mitochondrial DNA control region sequences of 91 domestic sheep from Kenya and found a high diversity of matrilines from the widespread haplogroup B, whereas only a single individual from haplogroup A was detected. Our phylogeography analyses of more than 500 available mitochondrial DNA sequences also identified ancestral haplotypes that were probably first introduced in Africa and are now widely distributed. Moreover, we found no evidence of an admixture between East and West African sheep. The presence of shared haplotypes in eastern and ancient southern African sheep suggests the possible southward movement of sheep along the eastern part of Africa. Finally, we found no evidence of an extensive introduction of sheep from southern Asia into Africa via the Indian Ocean trade. The overall findings on the phylogeography of East African domestic sheep set the grounds for understanding the origin and subsequent movements of sheep in Africa. The richness of maternal lineages in Kenyan breeds is of prime importance for future conservation and breeding programmes.     

Resolving ambiguities in the karyotype of domestic sheep (Ovis aries)

The unambiguous identification of ovine chromsomes has become essential for the mapping of the sheep genome, which predominantly consists of telocentric chromosomes of gradually decreasing size. Nucleolus organizer regions (NORs) and Robertsonian fusions have been used here as the cytological and morphological markers, respectively, to define the banding pattern of eight paris of sheep telocentric chromosomes that have an ambiguous identification status. Five Robertsonian chromosomes involving most of the ambiguous chromosomes as well as normal prometaphase chromosomes were stained sequentially and separately by QFQ, GTG, and Ag-NOR methodologies. The prometaphase banding patterns of the ambiguous chromosomes 4, 6, 8, 9, 21, 24, 25 and 26 are represented schematically. For providing an accurate image of the banding pattern, a system of shading has been employed to show the relative intensity of bands in a given chromosome. The results presented here will facilitate the regional mapping of the sheep genome, extend the information on cytogenetic homology with other bovids, and substantially accelerate the comparative mapping studies in Bovidae.     

Mechanics of locomotion of dogs (Canis familiaris) and sheep (Ovis aries)

Records have been made of the forces exerted on the ground by dogs and a sheep, in walking, trotting, cantering and slow galloping. Film has been taken simultaneously. The difference between walking and trotting was much less marked for the sheep than for the dogs.
Step length and stride length increase as speed increases. They are expressed as functions of the Froude number.
The vertical component of the force exerted by a foot on the ground shows two main maxima in walking, except in the case of the fore feet of sheep. In this case and in other gaits there is only one main maximum. The vertical movements of the fore and hind quarters which occurred in examples of each gait have been calculated from the force records.
The force exerted by a foot on the ground changes direction in the course of a step so as to remain more or less in line with a point fixed relative to the animal, but dorsal to its back.
The force records show impact disturbances in the first 003 sec of contact of each foot with the ground.
The point of application of the force on the sole of a foot tends to move posteriorly as the force increases.
The results are discussed in relation to a theoretical account of the mechanics of locomotion on legs.