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Genomic heterogeneity of small ruminant lentiviruses: existence of heterogeneous populations in sheep and of the same lentiviral genotypes in sheep and goats.

We have recently shown that French small ruminant lentiviruses (SRLV) from sheep are more similar to Caprine Arthritis Encephalitis Virus (CAEV) than to visna maedi virus (VMV) in a conserved region of the pol gene. To extend these results, we have examined sequences from a variable region of the env gene in French SRLV. We found that they were nearly equally distant from both CAEV and VMV strains, suggesting a considerable divergence since the initial introduction of the virus. Analysis of separate clones from individual animals showed that some carry a population of variant viruses. The study of further pol gene sequences from both goats and sheep suggests that viral variants show little or no host species specificity. A phylogenetic tree of pol gene sequences confirmed the presence of a novel genotype of SRLV in France.

Amino Acid Sequence↗

Sheep erythrocyte rosettes in pigs, sheep, cattle and goats demonstrated in the presence of dextran.

Formation of rosettes with sheep RBC by blood lymphocytes from young pigs was increased from 24.4% +/- 2.2 (mean +/- S.E.) in PBS to 54.1% +/- 1.9 in the presence of dextran. This increase was achieved without inducing appreciable rosette formation with other RBC which do not form rosettes in PBS. Lymphocytes which rosette only in dextran are predominant in pigs between 20 and 160 days old, when the peripheral lymphocyte pool is increasing very rapidly. The use of dextran revealed major populations of blood lymphocytes rosetting with SRBC in adult sheep (30.7% +/- 2.0), adult cattle (37.3% +/- 4.2) and adult goats (13.3% +/- 1.2). Proportions of rosette-forming lymphocytes tended to increase with age. In calf lymphoid tissues the distribution of rosette-forming lymphocytes suggested that these were T cells. In the improvement of rosette formation with SRBC, dextran was more effective than foetal calf serum, papain treatment of the SRBC or combinations of these treatments.

Age Factors↗

Comparison between in vitro lipid peroxidation in fresh sheep platelets and peroxidative processes during sheep platelet ageing under storage at 4 degrees C.

Incubation of sheep platelet crude membranes with xanthine oxidase (XO)/hypoxanthine/Fe(2+)-ADP revealed: (i) a fast peroxidative response - with a maximal linear rate of 14 nmol malondialdehyde (MDA) equivalents/mg protein, as evidenced by the thiobarbituric acid test - and a decrease in the polyunsaturated fatty acid (PUFA) content of the platelet crude membranes; (ii) a decrease in the lipid fluidity in the deep lipid core of the membranes but not at the membrane surface; (iii) a dramatic inhibitory effect on glucose 6-phosphatase (Glc-6-Pase) but not on acetylcholinesterase activity. Platelets were also aged by storage at 4 degrees C in their own plasma or in Seto additive solution. In these media, platelet aggregates were visible and the effects on platelet phospholipids, PUFA, lipid extract fluorescence, crude membrane fluidity and membrane-bound enzyme activities were assessed for comparison with those observed in in vitro lipid peroxidation. The sensitivity of membranes from stored platelets to lipid peroxidation was also assessed. Storage of platelets in plasma for 5 days was associated with different changes in their crude membranes such as decreases in arachidonic acid contents, the decrease not being avoided by the presence of phospholipase A(2) inhibitors, increases in MDA equivalents, conjugated dienes and lipid extract fluorescence, decreases in the amounts of MDA equivalents formed by platelet crude membranes treated with the oxidizing agents, changes in membrane fluidity and inhibition of Glc-6-Pase. All these alterations were less pronounced or even abolished after platelet storage in Seto. These findings suggest that platelet lipid peroxidation due to XO/hypoxanthine/Fe(2+)-ADP and platelet membrane alterations observed after platelet ageing under storage at 4 degrees C share common features. Also, as regards the prevention of peroxidative processes, Seto solution permits better storage of sheep platelets than plasma.

Acetylcholinesterase↗

Sulfur--selenium studies in sheep. II. Effect of a dietary sulfur deficiency on selenium and sulfur metabolism in sheep fed varying levels of selenomethionine.

The effect of a sulfur deficiency on the metabolism of selenium and sulfur was investigated in eight merino wethers. The sheep were fed high-sulfur (2 g S/kg) or low-sulfur (0.5 g S/kg) diets for two periods of 35 days each, and received selenium as selenomethionine at dietary concentrations of 0.02, 0.06, 0.09 and 0.67 mg Se/kg. Sheep fed the low-sulfur diet had reduced feed intake, reduced nitrogen, sulfur and selenium balance, but elevated plasma and wool selenium concentrations. Selenium concentrations in organs and tissues of slaughtered animals paralleled the selenium intake of the animal, with the renal cortex containing the highest concentration and bone the lowest. The effect of the 0.5 g S/kg diet on feed intake is in contrast with the results from the previous experiment (White and Somers 1977) using 0.7 g S/kg. It is this difference in fed intake which was responsible for many of the effects on selenium metabolism observed in this experiment. Once the feed intake effects are accounted for, the implications for sulfur--selenium interactions remain as before, i.e. more selenium is incorporated into wool and plasma protein when dietary sulfur is limiting than when it is not.

Animals↗

Air movement and heat loss from sheep. I. Boundary layer insulation of a model sheep, with and without fleece.

A model sheep, made from metal cylinders and hemispheres, was heated electrically. Heat loss by forced convection in a wind tunnel was analysed in terms of the dependence of the Nusselt number (Nu) on Reynolds number (Re). For a bare trunk Nu = 0.095 Re0.684, but with fleece covering the trunk to a depth of 3.5 cm, Nu = 0.0112 Re0.875 when the mean radiative temperature of the the coat was taken as the surface temperature. Heat transfer by convection from the whole body, including legs, was described by Nu = 0.029 Re0.80. However, a bulk Nesselt number should not be used to estimate heat loss from a live sheep in a hot environment if the windspeed is below about 4 m s-1 because the relation between mean surface temperature, Nusselt number and convective heat flux is not unique.

Air Movements↗

Relationships between the density of different indicator organisms on sheep and beef carcasses and in frozen beef and sheep meat.

AIM: To describe the relationship between the concentration of different indicator bacteria in red meat. METHODS AND RESULTS: Enumeration data for aerobic plate count (APC), Enterobacteriaceae, coliforms and Escherichia coli biotype I were analysed from an Australia-wide survey of beef carcasses, sheep carcasses, frozen beef and frozen sheep meat. In all commodities, there was only low-to-moderate rank correlation (0.16-0.47) between concentration of APC and concentration of each Gram-negative indicator. Rank correlations between counts of Gram-negative indicators were much higher (0.47-0.92) especially when nondetections were excluded from analysis (0.78-0.94). Receiver-operator characteristics analysis showed that detection of coliforms can predict the presence of E. coli biotype I with almost 100% sensitivity but fails to predict absence in 2.7-8.5% of samples not containing E. coli biotype I. CONCLUSIONS: Enumeration of coliforms is a useful adjunct to enumeration of E. coli biotype I or Enterobacteriaceae in red meat. The density of coliforms or Enterobacteriaceae can be used to predict the presence or absence of E. coli biotype I, although when the latter is at low prevalence errors in positive test prediction can be large. SIGNIFICANCE AND IMPACT OF THE STUDY: A quantitative basis is provided for comparing the concentration of different indicator bacteria measured in the production, regulation and trade of red meat.

Animals↗

Stimulation of Na+-K+ pump in sheep red blood cells by heteroimmune anti-sheep red cell antibodies.

In the HK-LK polymorphism of sheep red blood cells, alloimmune antiserum against the L antigen on LK cells is known to stimulate the Na+-K+ pump in low K+ (LK) cells, but alloimmune antiserum against the M antigen of high K+ (HK) cells does not. We have shown for the first time that heteroimmune antibodies against sheep red blood cells raised in mice can stimulate the pump. Heteroimmune antibodies against both LK(L) cells and HK(M) cells stimulated active K+ transport in LK cells. Furthermore heteroimmune antibodies against LK(L) cells also stimulated the pump in HK cells. As expected, alloimmune and heteroimmune antibodies acted at different sites in stimulating transport in LK cells.

Animals↗

Immunoglobulin-bearing lymphocytes: their demonstration in adult sheep and ontogeny in the sheep fetus.

A method for the preparation of lymphocytes from sheep blood is described. Lymphocytes from adult and fetal sheep have been examined by immunofluorescence for surface immunoglobulin (sIg) determinants. Young adults (1.5 years) had a mean of 23.2% sIg cells, older adults (8.75 years) a mean of 9.7% sIg cells. Pregnancy did not influence these values. The earliest lymphocytes with sIg in fetal lambs were demonstrable at 52 days (96 mm crown-rump length) and countable by 56 days (110 mm CRL) at 0.3% sIg. The percentage rose rapidly to 15.1% between 78 and 87 days, followed by a fall to 2.1% around 117 days with a second increase thereafter. The significance of this spontaneous appearance of fetal sIg cells is discussed.

Aging↗

Plasma norepinephrine, epinephrine, and dopamine concentrations in maternal and fetal sheep during spontaneous parturition and in premature sheep during cortisol-induced parturition.

The plasma catecholamine response to parturition was studied in two groups of chronically catheterized fetal sheep. Group I consisted of full term pregnancies with a mean gestation of 147 days at delivery. Group II was comprised of fetal sheep infused with cortisol to induce premature delivery at a mean gestational age of 135 days. Plasma catecholamine concentrations were measured in group I mothers and fetuses and group II fetuses by radioenzymatic assay using 50 microliter samples. Uterine contractions began 4--14 h before delivery in both groups. Significant increases in fetal plasma norepinephrine (NE) concentrations occurred within 3 h of delivery in both groups. Group I plasma epinephrine (E) and group I and II plasma dopamine levels were significantly increased within 1 h of delivery. Only plasma E levels differed in group I and II fetuses; there was no significant rise in plasma E concentrations in cortisol-infused (group II) fetuses. Maternal plasma NE and E concentrations were increased 2--3 h before delivery in group I animals. Maternal plasma dopamine concentrations were only elevated 30 min before delivery when plasma NE and E levels were highest.

Animals↗

[Estrus and ovulation synchronization in Merino meat sheep. 1. Effect of Gonavet "Berlin Chemie" after estrus synchronization with prostaglandin F2 alpha in Merino meat sheep].

Oestrus synchronisation by means of PGF2 alpha analogues was followed by injection of Gonavet "Berlin-Chemie" which triggered off an LH peak, 2 to 3 hours from injection. Injection of Gonavet "Berlin-Chemie", 44 hours after PGF2 alpha application, caused synchronisation of all LH peaks. The interval between injection of Gonavet "Berlin-Chemie" and onset of ovulation amounted to 22 hours. The length of ovulation was not accurately determinable. Ovulation was successfully induced to all sheep by application of Gonavet "Berlin-Chemie", 44 or 48 hours after PGF2 alpha injection. Ovulation rates were 1.75 or 1.54. Luteolytic action on sheep of Cloprostenol "Jenapharm", a PGF2 alpha analogue, proved to be just as good as that of Oestrophan (SPOFA).

Animals↗

Altered humoral immune response to sheep red blood cells by sheep erythrocyte-soluble hemolysate.

Adult C3H/He mice were rendered unresponsive to a primary injection of sheep red blood cells (SRBC) by pretreatment with sheep hemolysate supernatant (SHS) or subfractions of SHS isolated by column chromatography. The following effects of SHS on the immune response were observed: SHS did not kill antigen-reactive cells, it did not prevent the release of antibody by cells actively synthesizing and secreting antibody, and SHS-induced tolerance was not inhibited or abrogated by methods which terminate or abolish tolerance. In addition, cell-mediated responses were not affected in animals whose humoral responses were suppressed; however, the secondary plaque-forming cell (PFC) response was enhanced by SHS treatment. SDS gel electrophoresis revealed SHS to contain several proteins ranging from 12,000 to approximately 500,000 daltons.

Animals↗

Studies on the differentiation of T lymphocytes in sheep. I. Recognition of a sheep T-lymphocyte differentiation antigen by a monoclonal antibody T-80.

The results presented in this paper demonstrate that a mouse IgM monoclonal antibody (T-80) recognizes an antigen on cells of the T-lymphocyte lineage of sheep. However, this antibody does not identify all T cells, as 10-20% of thymocytes and some peripheral-blood T cells are negative. T-80- thymocytes reside in the medulla. The majority of cortical thymocytes are T-80+ and classified as dull cells on the basis of antigen density per cell as measured by flow microfluorometry. In contrast, T-80+ cells in the periphery can be categorized into two populations, i.e., dull cells and bright cells. Suggestive evidence was obtained that bright T-80+ cells are fast recirculating T cells, whereas dull cells are sessile or less easily mobilizable T cells in the periphery. In foetal environment, over 90% of thymocytes and approximately 5% of spleen cells are T-80+ at 54 days of gestation (gestation period = 150 days), which may indicate that T-cell emigration from the thymus commences well before mid-gestation in sheep.

Animals↗

Studies on the immune response to fixed antigens. Preferential induction of helper function with heavily trinitrophenylated sheep erythrocytes, and glutaraldehyde-treated sheep erythrocytes.

Mice primed with heavily trinitrophenylated sheep red cells (TNP128SRC) or glutaraldehyde-treated sheep red cells (G-SRC) developed an early helper function mediated by thymus-derived cells. Such mice were able to produce high secondary responses to both hapten and carrier after challenge 2 days after priming, with lightly trinitrophenylated SRC (TNP0.14SRC). However, the primary response of the TNP128SRC or G-SRC-primed mice were very low to undetectable, and their secondary responses were also low when the challenge antigen was administered 4 days after priming or later. Inhibitory humoral factor(s) which were induced in the primed animals appeared responsible for the decreased capacity of primed mice to mount a secondary response when challenged later than 2 days after priming. Transfer of spleen cells from TNP128SRC-primed mice to sublethally irradiated recipients circumvents their exposure to inhibitory humoral factor(s) present in intact animals allowing them to react with challenge antigen. Enriched populations of T cells, but not B cells, were able to transfer this early immunologic memory to irradiated recipients. The theoretical and practical implications of these results are discussed.

Aldehydes↗

Complexities in ETS-domain transcription factor function and regulation: lessons from the TCF (ternary complex factor) subfamily. The Colworth Medal Lecture.

The ETS-domain transcription factor family can be divided into a series of subfamilies. Elk-1 represents the founding member of the ternary complex factor (TCF) subfamily. By focusing on the TCF subfamily, we can demonstrate the complexities that exist in the function and regulation of ETS-domain transcription factors. This article focuses on Elk-1 in detail and summarizes the functions of other TCFs. The key themes covered include the domain structure of the TCFs, the mechanisms of complex formation with serum response factor, regulation of TCFs by mitogen-activated protein kinase cascades, and transcriptional regulatory properties of the TCFs. Finally, the emerging role of the TCFs in vivo is discussed. A picture is developing indicating that, while these proteins exhibit significant sequence and functional conservation, key differences in their structure and regulation are being identified which may relate to unique functions of these proteins in vivo.

Amino Acid Sequence↗

Higher relative, but lower absolute risks of myocardial infarction in women than in men: analysis of some major risk factors in the SHEEP study. The SHEEP Study Group.

OBJECTIVES: Middle-aged men have often been the subjects of multifactorial studies of myocardial infarction (MI) risk factors. One major objective of the SHEEP study was to compare the effects of different MI risk factors in women and men. DESIGN: SHEEP (Stockholm Heart Epidemiology Program) is a population-based case-referent study of causes of MI (first event) in Swedish women and men aged 45-70 years. During the period 1992-94, 2246 cases of MI were identified; 34% of the cases were women and 27% of the cases were fatal. One referent per case was chosen randomly from the Stockholm County population after stratification for the case's sex and age. Logistic regression was used to estimate the relative risks associated with risk factors of primary interest (diabetes, hypercholesterolaemia, hypertriglyceridaemia, hypertension, overweight, physical inactivity, smoking and job strain). RESULTS: The relative risk estimates ranged from 1.5 to 4.4 in women and from 1.3 to 2.9 in men (results for nonfatal cases and their referents). None of the 95% confidence intervals included 1.0. The relative risks were higher in the women than in the men (101-180%). The absolute risks, however, were all lower in the women than in the men. Estimates of Rothman's synergy index for gender ranged from 1.0 (hypertension) to 1.8 (current smoking). CONCLUSIONS: The indications of some effect modification due to sex (stronger risks in men for certain exposures) invoke the question of possible mechanisms.

Aged↗

Methionyl-tRNA synthetase from sheep liver. Purification of a fully active monomer derived from high-molecular-weight complexes by trypsin treatment. Evidence for immunological cross-reaction with the corresponding enzyme from sheep mammary gland.

The size distribution of methionyl-tRNA synthetase in extracts from sheep liver is compared to that of lysyl-tRNA, isoleucyl-tRNA, leucyl-tRNA and seryl-tRNA synthetases by gel filtration on Biogel A-5m. Extraction conditions are described which lead to isolation of methionyl-tRNA synthetase exclusively in the form of complexes of molecular weight close to 10(6). Limited trypsin treatment of these aggregates releases a fully active low-molecular-weight form of methionyl-tRNA synthetase which was purified to a specific activity of 674 units/mg at 25 degrees C with a yield of 40%. The homogeneous enzyme appears to be undistinguishable from the corresponding enzyme derived from sheep lactating mammary gland, as judged by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and by titration with antibodies raised against the enzyme purified from liver.

Amino Acyl-tRNA Synthetases↗

Culture of sheep Chlamydia in a sheep fibroblast cell culture.

Abortion and enteric isolates of Chlamydia psittaci from sheep differed in their growth in a fibroblastic cell culture derived from the small intestine of a lamb. Twenty abortion isolates, each from a different farm, produced large inclusions which could be passaged several times whereas 10 enteric isolates each from different farms (but from some of the farms of origin of the abortion isolates) produced sparse inclusions which could not be passaged. This appears to be a rapid method of distinguishing abortion and enteric isolates and may indicate different nutritional requirements or be related to the invasiveness of the isolates.

Abortion, Veterinary↗