Stimulatory effect of plasma samples from fattening cattle on adipogenesis‐related gene expression in preadipocyte cells

It is desirable to produce beef with high levels of monounsaturated fatty acids (MUFA), as this is related to fat softness and palatability. However, the physiology of MUFA synthesis in bovine fat during the fattening process remains to be established. In this study, in order to elucidate the relationship between plasma components and the fatty acid composition of intramuscular fat, we investigated the effect of plasma obtained from fattening cattle on the messenger RNA (mRNA) expressions of the adipogenesis‐related gene in a clonal bovine intramuscular preadipocyte line (BIP cells). The mRNA expressions of stearoyl‐CoA desaturase, adipocyte Protein 2, peroxisome proliferator‐activated receptor gamma and sterol regulatory element‐binding protein 1 in BIP cells were significantly higher following treatment with those plasma samples collected from the cattle with the highest diaphragmatic unsaturated fatty acids to saturated fatty acids ratio (US/S). Furthermore, the concentration of nonesterified fatty acid (NEFA) in the plasma samples had an inverse correlation with carcass diaphragmatic US/S. These results indicate that cattle with a low ratio of US/S in fat may be discriminated from the population of fattening cattle before slaughter by measuring the effect of their plasma on gene expression in BIP cells as well as their plasma concentration and composition of NEFA.     

Plasma vitamin C concentration is not related to the incidence of ketosis in dairy cows during the early lactation period

Many animals including cattle can synthesize vitamin C from glucose. The objective of this study was to investigate plasma vitamin C concentration in ketotic cows during the early lactation period because glucose supply for vitamin C synthesis might be limited in these cows. We measured plasma beta-hydroxybutyrate (BHBA) concentration in 118 cows within 2 months after parturition. Subclinical/clinical ketosis was quantitatively determined using a plasma BHBA threshold of 1,200 microM. Plasma glucose concentration was lower in the ketotic cows than in the control cows but plasma vitamin C concentration did not differ between the control and the ketotic cows. Then we measured plasma vitamin C, BHBA and glucose levels in 7 cows during the periparturient period. Plasma BHBA increased and plasma glucose decreased after parturition but plasma vitamin C did not change. These results indicate that plasma vitamin C is not related to the incidence of ketosis in the early lactation period. We suggest that ketotic cows have the ability to produce vitamin C to meet its requirement in the early lactation period although glucose supply is not sufficient for milk production. Vitamin C synthesis is possibly given a high metabolic-priority for glucose in lactating cows.     

Antiradical function of sulfhydryl residues and oxidative stress markers in dairy cattle plasma

The antiradical function of sulfhydryl (SH) residue in dairy cattle plasma and the relationship of SH residue concentrations to other oxidative stress markers, ascorbic acid and thiobarbituric acid reactive substance (TBARS) in plasma were investigated. The concentration of reduced glutathione (GSH) in phosphate buffer solution (pH 7.6) and SH residues in dairy cattle plasma decreased significantly (P < 0.05) in vitro by the addition of peroxyradicals at 38°C, depending on incubation periods. The decrease of GSH concentration with the peroxyradical solution was partially protected by the addition of sodium ascorbate solution. A positive and significant correlation with SH residues and albumin concentration in the fresh plasma obtained from 15 dairy cattle was observed (P < 0.05). The SH residue concentration was not correlated with the TBARS concentration in plasma. The total ascorbic acid and SH residues concentration in the plasma correlated positively but not significantly (P < 0.10). These results suggested that SH residues in dairy cattle plasma play important part in the antiradical function.     

The effect of vitamin C supplementation on plasma concentration and urinary excretion of vitamin C in cattle

We investigated the plasma concentration and urinary excretion of vitamin C in cows supplemented with vitamin C. Five cows (mean BW = 597 kg) were allocated to a 5 x 5 Latin square design and supplemented with a vitamin C preparation coated with hydrogenated soybean oil at 0, 10, 20, 40, or 60 mg of vitamin C per kg of BW per day for 9 d. Plasma and urine samples were collected for measuring vitamin C concentration. Urinary excretion of vitamin C was expressed as the ratio of vitamin C to creatinine. Plasma vitamin C concentration and urinary vitamin C excretion increased quadratically as dietary vitamin C increased (P < 0.001); that is, the lowest dose affected neither plasma vitamin C concentration nor urinary vitamin C excretion but the plasma vitamin C concentration and urinary vitamin C excretion increased (P < 0.05) with increasing supplementation of vitamin C at greater doses. This suggests that plasma vitamin C concentration affects urinary excretion of vitamin C in cattle and that plasma vitamin C concentration exceeded the renal threshold for vitamin C in the cows receiving vitamin C at 20 mg/kg of BW per day. Furthermore, increased urinary excretion of vitamin C appears to limit plasma vitamin C concentration in response to vitamin C intake. The daily excretion of vitamin C was estimated by the reported value of daily creatinine excretion, indicating that the daily amount of vitamin C excreted into urine was more than half of supplied vitamin C. Therefore, a large part of supplied vitamin C probably escapes ruminal degradation and is absorbed but excreted into urine.     

Effects of vitamin C supplementation on plasma ascorbic acid and oxalate concentrations and meat quality in swine

Two experiments were conducted to determine the effects of vitamin C supplementation 48 h before slaughter on plasma ascorbic acid and oxalate concentrations and its effect on pork quality. In Exp. 1, 16 pigs (87.8+/-2.13 kg BW) were blocked by sex and weight and assigned randomly within block to one of three vitamin C treatments: 1) control; 2) 1,000 mg/L; or 3) 2,000 mg/L supplemented in the drinking water for a 48-h period. This was then followed by an additional 48-h period without supplemental vitamin C. Vitamin C increased plasma ascorbic acid concentrations (11.6, 19.5, and 23.4 microg/mL for 0, 1,000, and 2,000 mg/L of vitamin C; P < 0.05) within 6 h of supplementation. Plasma ascorbic acid concentrations from treated pigs decreased and did not differ from those of control pigs (13.7, 18.2, and 18.6 microg/mL for 0, 1,000, and 2,000 mg/L of vitamin C; P = 0.30) within 2 h of ending supplementation. No differences in plasma ascorbic acid concentrations were found between the two levels of supplementation. Vitamin C did not affect plasma oxalate or cortisol; however, cortisol tended to increase quadratically (P = 0.077) with vitamin C after 96 h. In Exp. 2, 30 pigs (107.5+/-0.54 kg BW) were blocked by sex and weight and assigned randomly within block to one of three vitamin C treatments: 1) control; 2) 500 mg/L; or 3) 1,000 mg/L supplemented in the drinking water 48 h before slaughter. Pigs were slaughtered 4 to 5 h after vitamin C supplementation ended, and loin samples were collected for meat quality measurements. At the time of slaughter, no differences in plasma ascorbic acid or cortisol were observed, but oxalate tended (P = 0.074) to increase quadratically with increasing vitamin C. Muscle ascorbic acid at slaughter and lactic acid in muscle at 0 and 1.5 h after slaughter were not different; however, lactic acid increased (P = 0.048) quadratically at 24 h after slaughter. Vitamin C did not affect initial or ultimate pH. Initial fluid loss (P = 0.041), and fluid loss on d 4 (P = 0.014) and 8 (P = 0.076) of simulated retail display; L* on d 0 (P = 0.038), 4 (P = 0.010), and 8 (P = 0.051); a* on d 0 (P = 0.021); and b* on d 0 (P = 0.006), 4 (P = 0.035), and 8 (P = 0.017) were negatively affected in a quadratic manner when vitamin C was supplemented. Vitamin C tended (P = 0.086) to increase oxidation in chops on d 0, but not d 4 or 8. Results indicate that on-farm supplementation of vitamin C was generally not effective in improving pork quality, which may be related to timing relative to slaughter.     

An automated micro-method for the enzymatic determination of D-B-hydroxybutyrate in ruminant plasma

1. An enzymatic ‘reaction rate’ micro-method for the rapid routine estimation of D-B-hydroxybutyrate (D-B-HOB) in ruminant plasma, using an I.L. Multistat III centrifugal analyzer, is described.
2. Reaction conditions were optimized to give a linear response for plasma D-B-HOB concentrations between 100 and 2500 μmoles per litre, at 30°C and pH 9.0.
3. For the standardized method the within-run and between-run coefficients of variation for deproteinised ovine plasma were consistently less than 3.5%.
4. There was good agreement between plasma concentrations obtained by the present method and both original U.V. end-point technique (r=0.927b=0.950) and a colorimetric end-point procedure (r=0.937. b=0.879).
5. Utreated ovine and bovine plasma consistently exhibited high ‘blank’ activity and this was directly correlated with plasma lactate dehydrogenase (LDH) activity in both species (r=0.971; p<0.001 and r=0.949; p<0.001 respectively). The distribution of LDH activity in man was similar to sheep but, contrastingly, non-specific interference was extremely low in human plasma and unrelated to LDH. Horse, chicken and rat had negligible ‘blank” activity and comparatively low LDH levels. In both cattle and sheep non-specific interference was abolished by perchloric acid precipitation. In the sheep subtraction of ‘blank’ activity gave D-H-HOB concentrations for untreated plasma comparable to those in deproteinised samples. However, in the bovine, D-B-HOB levels remained significantly (t=6.44; p<0.001) higher even after ‘blank’ correlation. In contrast to man and other non-ruminants, perchloric acid precipitation is essential in ruminants to avoid false overestimation of plasma D-B-HOB levels.
6. Plasma with EDTA as anticoagulant and serum gave concentrations of D-B-HOB approximately 60% lower, than samples containing heparin or oxalate/fluoride. However, heparin was associated with much higher (up to 50%) non-specific NAD rduction than oxalate/fluoride.
7. High levels of acetoacetate (400–1000 μmoles per litre) reduced the recovery of D-B-HOB from ovine plasma by less than 10%. This effect was negated by the inclusion of hydrazine hydrates in the reaction mixture. Perchlorate ion concentrations above 25 μmoles per litre per test dramatically inhibited the assay in ovine plasma, and therefore precipitation conditions must be carefully controlled.
8. Plasma with oxalate/fluoride as anticoagulant showed the greatest stability in storage; 24 hours at room temperature, one week at +4°C and at least one month at ?20°C.

     

Calculation of variables describing plasma nonvolatile weak acids for use in the strong ion approach to acid-base balance in cattle

OBJECTIVE: To calculate values for the total concentration of nonvolatile weak acids (Atot) and the effective dissociation constant for nonvolatile weak acids (Ka) of bovine plasma and to determine the best method for quantifying the unmeasured strong anion concentration in bovine plasma. SAMPLE POPULATION: Data sets from published and experimental studies. PROCEDURE: The simplified strong ion model was applied to published and experimentally determined values for pH, PCO2, and strong ion difference (SID+). Nonlinear regression was used to solve simultaneously for Atot and Ka. Four methods for quantifying the unmeasured strong anion concentration in plasma (anion gap, the Fencl base excess method [BEua], the Figge unmeasured anion method [XA], and the strong ion gap [SIG]) were compared in 35 cattle with abomasal volvulus. RESULTS: For bovine plasma at 37 C, Atot was 25 m M/L, equivalent to 76 times the albumin concentration or 3.6 times the total protein concentration; Ka was 0.87 x 10(-7), equivalent to pKa of 706. The Atot and Ka values were validated, using data sets from in vivo and in vitro studies. Plasma unmeasured strong anion concentration was most accurately predicted in critically ill cattle by calculating SIG from serum albumin (R2, 0.66) or total protein concentration (R2, 0.60), compared with BEua (R2, 0.56), [XA] (R2, 0.50), and the anion gap (R2, 0.41). CONCLUSIONS AND CLINICAL RELEVANCE: Calculated values for Atot, Ka, and the SIG equation should facilitate application of the strong ion approach to acid-base disturbances in cattle.     

The Relationship Among Vitamin C, β‐carotene,Vitamin A,Progesterone and Oestradiol 17‐β Concentrations in Plasma and Cyst Fluid of Holstein Cows with Ovarian Cyst

The aims of this study were to determine the concentrations of the progesterone, oestradiol‐17‐β, vitamin A, C and β‐carotene in plasma and cyst fluid and to relate these values with cystic diameter and membrane thickness of Holstein cattle with ovarian luteal cyst. 1650 Holstein cows were examined for the presence of the ovarian cyst and luteal and follicular cystic ovaries were obtained following slaughtering in personal slaughterhouse in Konya‐Turkey. 15 Luteal and 15 follicular cystic ovaries were distinguished by rectal palpation and by post mortem ultrasonographic examination. Plasma and cyst fluid, hormone and vitamin analyses were carried out by EIA method and spectrophotometric measurement respectively. Although there was no relationship between β‐carotene and vitamin A in plasma and cyst fluid of both cyst type and hormone concentrations, the vitamin C concentration of cyst fluid was found significantly higher in luteal cyst than in follicular cyst. Moreover, there is a positive correlation among values of the vitamin C concentrations of cyst fluid and cystic membrane thickness, plasma and the cyst fluid progesterone concentrations, but there is a negative correlation among the vitamin C concentrations of cystic fluid and oestradiol 17β levels of plasma and cyst fluid. In conclusion, vitamin C concentration of cyst fluid supported ultrasonographic and endocrinologic findings. Also, it can be postulated that vitamin C is probably effective on progesterone synthesis in the luteal tissue of cyst.     

Effect of high environmental temperatures on ascorbic acid, sulfhydryl residue and oxidized lipid concentrations in plasma of dairy cows

Information on oxidative stress under hot conditions from the levels of cells to organs and the whole body has accumulated in the last decades. Although a hot climate decreased dairy performance, changes of oxidative stress markers under hot conditions have remained obscure. Therefore, the effect of high environmental temperature on ascorbic acid, sulfhydryl (SH) residue and oxidized lipids concentrations in plasma from a total of 128 dairy cows was investigated. The monthly average maximum day temperature varied from 9.2°C in January to 32°C in August of 2004 in this institute. High ambient temperatures increased the rectal temperature of dairy cows up to 39.3°C in August. One of the reducing equivalents in plasma, SH residue concentration, decreased in July compared with December (P < 0.05). Another antiradical molecule, ascorbic acid concentration in plasma, also decreased in July (P < 0.01). The oxidative stress index, thiobarbituric acid reactive substance (TBARS), which was produced from the oxidation of polyunsaturated fatty acids under oxidative conditions, increased in summer (P < 0.05). A significant positive relationship of SH residue and ascorbic acid concentrations in the hot season was observed (P < 0.01). A negative correlation between rectal temperatures and ascorbic acid concentrations in the hot season was obtained (P < 0.01). However, TBARS concentration varied independently of the SH residue and ascorbic acid concentration. These results suggest that the response of oxidative stress markers of SH residue, ascorbic acid and TBARS concentration to oxidative stress under hot conditions were not shown to be the same, and that oxidative stress in dairy cows in the hot season increased.     

Effects of injectable vitamins A, D, E and C on the health and growth rate of feedlot cattle destined for the Australian domestic market

OBJECTIVE: To examine the effects of injectable vitamins A, D and E at feedlot entry on health and growth rate and the effects of injectable vitamin C at the time of treatment for bovine respiratory disease (BRD) on cattle health. DESIGN: Systematic allocation of 2465 cattle at feedlot entry to: a commercial vitamin A, D and E preparation at the label dose rate; commercial vitamin A, D and E at twice the label dose rate; a formulation with no vitamin D, a lower concentration of vitamin A and a higher concentration of vitamin E; and the oil-based carrier alone at volumes corresponding to the above treatments. Comparisons of growth rate, disease and mortality were made between the groups at the conclusion of the feeding period. In a separate experiment, 176 cattle were alternately administered injectable vitamin C at the time of treatment for BRD, or were not injected with vitamin C, and mortality was compared between the groups. RESULTS: There were no differences between cattle administered vitamin A, D and E at feedlot entry and the controls in growth rate (P=0.11), all diseases (P=0.99), BRD (P=0.60) or mortalities (P=0.95). Cattle treated with the higher vitamin E and lower vitamin A preparation had a higher (P=0.02) incidence of anorexia than the other groups. Fewer cattle treated with 5 g of vitamin C by intramuscular injection at the time of treatment with antibiotics for BRD subsequently died (P=0.04). CONCLUSIONS: The routine injection of cattle with vitamins A, D and E at feedlot entry is unlikely to result in improvements in health and growth rate where cattle are provided with these vitamins in their diets at concentrations equal to the National Research Council recommendations. Mortality rate in cattle diagnosed with BRD may be reduced by intramuscular injection of vitamin C at the time of treatment with antibiotics.     

Preliminary system of rapid analysis of blood retinol level in cattle

Control of blood retinol levels in cattle during fattening is important in the production of marbled beef. However, it is difficult to easily measure the blood retinol concentration in the field. In this study, we attempted to develop an analysis method that does not require blood cell separation and uses a compact fluorescence analyzer that can be carried around as a preliminary system for measuring blood retinol concentration in the field. This system was used to monitor blood retinol levels in 12 fattening cattle (14 to 27 months old) and demonstrated a strong correlation (r = 0.78) with the results obtained by the standard high-performance liquid chromatography (HPLC) method. Stronger correlations (r = 0.87) were obtained until the cattle were 24 months of age. These results suggest that higher correlations can be expected to be obtained by improving the robustness of the extraction system. Refinements for practical use need to be considered, but whole blood extraction and the vitamin A analyzer that was developed show potential to be used for on-farm monitoring of retinol levels.     

Plasma concentration of some vitamins in greyhounds

Plasma concentrations of ascorbic acid, (3-carotene, retinol and a-tocopheral were determined in four groups of racing greyhounds. Plasma biotin was measured in two groups. Mean concentrations were 5-3 mg/litre ascorbic acid, 877|xg/litre retinol, 10-4 mg/litre a-tocopherol and 451 ng/litre biotin. In all dogs (3-carotene was below the level of detection, ie, 20 μg/litre. Differences in plasma vitamin concentrations between groups were found.     

Ascorbic acid status of female camels during different phases of reproduction

There is suggestive evidence that a low status of ascorbic acid in camels enhances their risk for infectious diseases. This study was carried out to disclose the role of reproduction, if any, in affecting ascorbic acid status. The associations between the reproductive cycle and ascorbic acid contents in plasma and leukocytes were studied in Sudanese camels browsing on local vegetation. Ascorbic acid status was found to be lowest during pregnancy and highest during lactation. Estrus versus non-estrus was associated with high vitamin C status. Brucellosis-positive camels showed decreased levels of ascorbic acid in plasma and leukocytes. Possibly, the phases of non-estrus and pregnancy in camels invoke an increased susceptibility to infectious diseases due to a lower ascorbic acid status.     

Mitogen stimulation of lymphocytes in pigs with hereditary vitamin C deficiency

Recently an inherited vitamin G deficiency in the pigs presumably based on an autosomal recessive gene was decribed* Homozygotes are in contrast to heterozygotes and normal pigs unable to synthesize ascorbic acid. In an experiment comprising 3 littermate pigs, 2 homozygous and 1 heterozygous for the vitamin C deficiency gene, the influence of ascorbic acid depletion, and repletion on mitogen stimulation of peripheral blood lymphocytes was studied. Ascorbic acid depletion of the vitamin C dependent pigs resulted in a rapid decline in plasma ascorbic acid. Response of lymphocytes to stimular tion with Concanavalin A (Con A) and phytohemagglutinin M (PHA) decreased more slowly reaching a minimum, which coincidedi with the occurrence of the first clinical symptoms of scurvy. Following resupplementation with vitamin C the plasma content of ascorbic acid rapidly returned to normal, while the lymphocyte response to Con A and PHA stimulation only gradually approached the initial values. The repletion with ascorbic acid caused a transitory increase in the response to pokeweed mitogen (PWM) stimulation. The significance of these findings in relation to the cellular immune system in normal pigs is discussed.     

Plasma leptin, feed intake and body fat accumulation in fattening castrated male and female lambs

The main objective of this study was to investigate the relationships between changes in plasma leptin concentration and feed intake or bodyweight in female and castrated male lambs with fattening. Four female and four castrated male lambs were used and were fed roughage and concentrate supplemented with beef tallow ad libitum for 28 weeks. Although the feed intake and bodyweight increased with fattening in both the castrated male and female lambs, they decreased at 24–28 weeks in the female lambs. At the end of fattening, the crude fat content in the muscle (loin) of the female lambs was significantly higher than in the castrated male lambs (P < 0.05), while the crude protein content in the loin and fillet meat was higher in the castrated male than in the female lambs (P < 0.05). The plasma leptin concentration showed high values at a later stage of fattening (P < 0.05). In the female lambs the plasma insulin concentration increased at a later stage of fattening (P < 0.05) and was positively correlated (P < 0.0001, r = 0.78) with plasma leptin. Plasma metabolites (glucose, nonesterified fatty acid, total cholesterol and triglyceride) concentrations were also changed with fattening. Plasma total cholesterol was positively related to plasma leptin, more closely in the female than in the castrated male lambs (in females, r = 0.63, P < 0.001; in males, r = 0.38, P < 0.01). The accumulation of body fat was probably accelerated by the consumption of a lot of concentrate feed supplemented with treated beef tallow and by the stimulation of insulin with fattening. Consequently, the plasma leptin concentration increased, especially toward the end of the fattening period. The decrease in feed intake and bodyweight after the 24th week of fattening was possibly caused by an increase in leptin that is involved in the homeostatic regulation of body energy by regulating appetite.     

Contribution of cobalamin analogues to plasma vitamin B12 concentrations in cattle

Plasma vitamin B12 concentrations in cattle were analysed by a radioisotope dilution assay using pig intrinsic factor and a microbiological assay using Euglena gracilis. Both assays provided similar results for samples of cattle plasma containing vitamin B12 concentrations ranging from 0.07 to 3.60 micrograms litre-1 (r = 0.95, P less than 0.001). The addition of excess cobinamide in the radioisotope dilution assay to block non-specific binding in the intrinsic factor preparation due to the presence of R-type binders, was used to determine the presence of cobalamin analogues. Cobalamin analogues accounted for up to 50 per cent of the total vitamin B12 concentration in samples of plasma from cows but were virtually undetectable in plasma from sheep.     

Sexual difference in ascorbic acid synthesis, tissue ascorbic acid and plasma total antioxidant capacity in mature chickens

1. An experiment was conducted with commercial White Leghorn type chickens to determine the effect of gender on tissue ascorbic acid concentration, antioxidant capacity and ascorbic acid synthesis. 2. Birds reared and maintained on litter were given a standard layer diet, without supplemental ascorbic acid, from 18 weeks of age. Tissue ascorbic acid concentration, plasma total antioxidant capacity and renal L-gulonolactone oxidase activity were measured at 30 weeks of age. 3. Females and males differed in ascorbic acid synthesis, as measured by renal L-gulonolactone oxidase activity, and tissue ascorbic acid concentration. 4. Plasma total antioxidant capacity and adrenal, gonadal, plasma and pituitary ascorbic acid concentrations were significantly higher in males, whereas ascorbic acid synthesis and splenic and thymic ascorbic acid concentrations were significantly higher in females. 5. L-Gulonolactone oxidase activity was not detected in the comb of cockerels.     

Effects of dietary supplemental L-carnitine and ascorbic acid on performance, carcass composition and plasma L-carnitine concentration of broiler chicks reared under different temperature.

The present study was initiated to determine whether dietary supplemental L-carnitine and ascorbic acid affect growth performance, carcass yield and composition, abdominal fat and plasma L-carnitine concentration of broiler chicks reared under normal and high temperature. During the experiment, two temperature regimes were employed in two experimental rooms, which were identical but different in environmental temperature. The regimes were thermoneutral (20-22 degrees C for 24 h) or recycling hot (34-36 degrees C for 8 h and 20-22 degrees C for 16 h). One-day-old broiler chicks (ROSS) were used in the experiment. A 2 x 2 x 2 factorial arrangement was employed with two levels (0 and 50 mg/kg) of supplemental L-carnitine and two levels (0 or 500 mg/kg) of supplemental ascorbic acid in drinking water under thermoneutral or high temperature regimes. Body weight gain was affected by high temperature. However, body weight gain was significantly improved in animals receiving supplemental L-carnitine, ascorbic acid or L-carnitine + ascorbic acid compared to animals receiving unsupplemented diet under high temperature. On the other hand, supplemental L-carnitine or L-carnitine + ascorbic acid reduced body weight gain under thermoneutral condition. Supplemental ascorbic acid significantly improved feed conversion efficiency, the improvement was relatively greater under high temperature. The L-carnitine content in the plasma was higher in the groups receiving supplemental L-carnitine and ascorbic acid under high temperature, while broilers fed supplemental L-carnitine and ascorbic acid had a decreased level of plasma L-carnitine concentration under normal temperature. It is concluded that dietary supplemental L-carnitine or L-carnitine + ascorbic acid may have positive effects on body weight gain, carcass weight under high temperature conditions.     

An automated micro-method for the enzymatic determination of D-B-hydroxybutyrate in ruminant plasma

An enzymatic 'reaction rate' micro-method for the rapid routine estimation of D-B-hydroxybutyrate (D-B- HOB ) in ruminant plasma, using an I.L. Multistat III centrifugal analyzer, is described. Reaction conditions were optimized to give a linear response for plasma D-B- HOB concentrations between 100 and 2500 mumoles per litre, at 30 degrees C and pH 9.0. For the standardized method the within-run and between-run coefficients of variation for deproteinised ovine plasma were consistently less than 3.5%. There was good agreement between plasma concentrations obtained by the present method and both original U.V. end-point technique (r = 0. 927b = 0.950) and a colorimetric end-point procedure (r = 0.937. b = 0.879). Untreated ovine and bovine plasma consistently exhibited high 'blank' activity and this was directly correlated with plasma lactate dehydrogenase (LDH) activity in both species (r = 0.971; p less than 0.001 and r = 0.949; p less than 0.001 respectively). The distribution of LDH activity in man was similar to sheep but, contrastingly , non-specific interference was extremely low in human plasma and unrelated to LDH. Horse, chicken and rat had negligible 'blank' activity and comparatively low LDH levels. In both cattle and sheep non-specific interference was abolished by perchloric acid precipitation. In the sheep subtraction of 'blank' activity gave D-B- HOB concentrations for untreated plasma comparable to those in deproteinised samples. However, in the bovine, D-B- HOB levels remained significantly (t = 6.44; p less than 0.001) higher even after 'blank' correction. In contrast to man and other non-ruminants, perchloric acid precipitation is essential in ruminants to avoid false overestimation of plasma D-B- HOB levels. Plasma with EDTA as anticoagulant and serum gave concentrations of D-B- HOB approximately 60% lower, than samples containing heparin or oxalate/fluoride. However, heparin was associated with much higher (up to 50%) non-specific NAD reduction than oxalate/fluoride. High levels of acetoacetate (400-1000 mumoles per litre) reduced the recovery of D-B- HOB from ovine plasma by less than 10%. This effect was negated by the inclusion of hydrazine hydrate in the reaction mixture. Perchlorate ion concentrations above 25 mumoles per litre per test dramatically inhibited the assay in ovine plasma, and therefore precipitation conditions must be carefully controlled. Plasma with oxalate/fluoride as anticoagulant showed the greatest stability in storage; 24 hours at room temperature, one week at +4 degrees C and at least one month at -20 degrees C.     

Content of beta-carotene, vitamin E and ascorbic acid in blood plasma of female calves, cattle, bulls, castrates and ox throughout the course of the year]

The concentration of beta-carotin, of vitamin E and of ascorbic acid in the plasma of female calves (beginning with an age of 4 weeks), of cows in the 2nd and 5th lactation, of bulls, of short scrotum bulls and of oxes in the course of the year was analysed. The concentration of beta-carotin in the plasma of calves after driving up to pasture increased slower than that of cows. After transition to stable feeding there was a significant decrease in the concentration after 4 weeks. During the period of suckling the concentration of vitamin E in the plasma of calves was low. After transition to pasture the concentration of vitamin E in the plasma of calves increased slower than in that of the cows. The concentration of ascorbic acid in the plasma during the winter was higher than during the summer. There was no influence of experimental cryptorchism or castration on the mentioned values.