Effect of blood lactate-guided conditioning of horses with exercises of differing durations and intensities on heart rate and biochemical blood variables.

The velocity at which blood lactate concentration ([LA]) of 4 mmol/L is reached (v(4)) is widely used to determine fitness, but there are few published data on using [LA] as a guide for the exercise speed for training in horses. In this study, the effect of 3 conditioning programs with [LA] guided exercise speeds on v(4), v(200) (speed at a heart rate of 200 beats/min), blood [LA], plasma FFA ([FFA]), and alanine concentrations ([alanine]), before and after exercise, as well as heart rate during exercise, of horses was examined. Six 2-yr-old Haflinger stallions underwent an initial treadmill-based standard exercise test (SET). A regression analysis [LA]-speed relationship was used to calculate v(1.5), v(2.5), and v(4). Horses were then randomly assigned to 1 of 3 conditioning programs according to a 3 x 6 Latin square design. During 6 wk, horses exercised on the treadmill every other day for 45 min at their calculated v(1.5) or v(2.5) or 25 min at their v(4). Each conditioning period (CDP) was followed by 5 wk without conditioning. At 2 and 9 d, and 5 wk, after the end of the CDP, the horses performed another SET to evaluate again the v(4) and v(200). Blood [LA], plasma [FFA], and [alanine] were measured before and after heart rate during exercise sessions 1, 11, and 21 in each CDP. None of the exercise programs had an effect on v(4) and v(200) (P > 0.05). The increase of the mean [LA] after exercise decreased during CDP (P < 0.05), and the increase of mean heart rate during exercise tended to decrease as well (P = 0.07). There was no difference among the conditioning programs. Plasma [FFA] before exercise was not influenced by the CDP (P > 0.05). The plasma [FFA] was always greater after exercise (P < 0.05), but there was no difference among conditioning programs. Overall, the increase was greatest after the 21st exercise session compared with the 1st and 11th exercise sessions (P < 0.05). The mean plasma [alanine] before exercise remained similar during all CDP (P > 0.05). Mean plasma [alanine] of the horses was increased after all exercise sessions measured (P < 0.05). There was no difference among conditioning programs (P > 0.05). It is concluded that conditioning with the exercise types used had small effects. This could have been because the exercise stress was too small, but also because the workload was not increased during the CDP.

The effects of dietary phosphorus deficiency on surface pH and membrane composition of the mucosa epithelium in caprine jejunum.

In ruminants, the uptake of inorganic phosphate (P(i)) across the intestinal mucosa epithelium by Na-dependent and Na-independent mechanisms is a main regulatory factor in P homeostasis. The aim of the study was to elucidate to which extent Na-independent mechanisms, including pH effects or composition of mucosal brush-border membranes, could be involved in positive stimulation of P(i) absorptive processes seen under the P deficient condition. Therefore, luminal, surface and intracellular pH of the jejunal epithelial cells in control and P depleted goats were compared and biochemical analyses of membrane phospholipids in the apical membrane of the jejunal epithelium were performed. Dietary P depletion resulted in decreased plasma P(i) levels. While pH in jejunal ingesta was not significantly changed, P depletion resulted in a significantly lower surface pH in the crypt region compared to control animals (7.62 +/- 0.02 vs. 7.77 +/- 0.04, n = 4, P < 0.01). Inhibition of apical Na(+)/H(+)-exchange resulted in an increase of the jejunal surface pH in P depleted animals by 0.07 +/- 0.01 (n = 6, P < 0.01) and 0.05 +/- 0.01 (n = 6, P < 0.01) for the villus and the crypt region, respectively. This increase were inversely correlated with the initial surface pH prior to inhibition. In contrast to surface pH, intracellular pH of the jejunal epithelium and the phospholipid composition of the apical jejunal membrane were not affected by P depletion. Although the data suggest the existence of a Na(+)/H(+)-exchange mechanism at the luminal surface of goat jejunum they do not support the hypothesis that adaptational processes of active P(i) absorption from goat jejunum in response to low dietary P could be based on "non P(i) transporter events".

Distribution patterns of the glucose transporters GLUT4 and GLUT1 in skeletal muscles of rats (Rattus norvegicus), pigs (Sus scrofa), cows (Bos taurus), adult goats, goat kids (Capra hircus), and camels (Camelus dromedarius).

Earlier studies demonstrated that forestomach herbivores are less insulin sensitive than monogastric omnivores. The present study was carried out to determine if different distribution patterns of the glucose transporters GLUT1 and GLUT4 may contribute to these different insulin sensitivities. Western blotting was used to measure GLUT1 and GLUT4 protein contents in oxidative (masseter, diaphragm) and glycolytic (longissimus lumborum, semitendinosus) skeletal muscle membranes of monogastric omnivores (rats and pigs), and of forestomach herbivores (cows, adult goats, goat kids, and camels). Muscles were characterized biochemically. Comparing red and white muscles, the isocitrate dehydrogenase (ICDH) activity was 1.5-15-times higher in oxidative muscles of all species, whereas lactate dehydrogenase (LDH) activity was 1.4-4.4-times higher in glycolytic muscles except in adult goats. GLUT4 levels were 1.5-6.3-times higher in oxidative muscles. GLUT1 levels were 2.2-8.3-times higher in glycolytic muscles in forestomach herbivores but not in monogastric animals. We conclude that GLUT1 may be the predominant glucose transporter in glycolytic muscles of ruminating animals. The GLUT1 distribution patterns were identical in adult and pre-ruminant goats, indicating that GLUT1 expression among these muscles is determined genetically. The high blood glucose levels of camels cited in literature may be due to an "NIDDM-like" impaired GLUT4 activity in skeletal muscle.

Assessment of the antioxidative potential of dietary supplementation with α-tocopherol in low-nitrite salami-type sausages.

The objective of this study was to determine the effects of dietary α-tocopherol supplementation to pigs on the lipid stability of nitrite-reduced salami-type sausages (custom-produced for this study). Fourteen crossbred pigs were divided into two equal groups. The control group (CG) was fed a basal diet containing 34ppm α-tocopherol, and the experimental group (EG), a diet supplemented with 364ppm α-tocopherol, both for 35 days. After slaughter, salami-type sausages were produced with the addition of different nitrite levels in the curing salt (100, 50, 25 and 0ppm), ripened for four weeks, sliced, and packaged under a protective atmosphere (80 parts N(2) to 20 parts CO(2)). The packed products were stored for eight weeks under simulation of the usual conditions in self-service shelves with permanent lighting (9°C, 200 lux). The sausages were regularly assessed for their oxidative status (thiobarbituric acid-reactive substances, fatty acid pattern) and the amount of α-tocopherol, and by sensory evaluation. In the EG sausages, α-tocopherol concentrations were twice as high and TBARS were lower than in the CG sausages. The α-tocopherol in EG prevented the loss of unsaturated fatty acids during storage. Sausages of EG containing 100ppm nitrite had the longest sensory stability. However, the biochemical parameters did not indicate that nitrite had any effect on the antioxidative potential of the sausages.

[Contents of the glucose transporters GLUT1 and GLUT4 in oxidative and glycolytic muscles of goat kids and adult goats]. / Gehalte der Glucosetransporter GLUT1 und GLUT4 in oxidativer und glycolytischer Muskulatur von Ziegenlammern und adulten Ziegen.

The objective of this study was to elucidate, whether the impaired insulin sensitivity with regard to glucose utilisation in ruminating goats compared with suckling goat kids may be due to a reduced expression of the glucose transporters GLUT1 and GLUT4 in skeletal muscle. Muscle samples were removed from red, oxidative muscles (M. masseter, diaphragm) and white, glycolytic muscles (M. longissimus dorsi, and M. semitendinosus) of five goat kids fed with milk exchanger and nine adult goats of different stages of life. Samples were analysed for their GLUT1 and GLUT4 contents. The muscles were characterised metabolically by measuring the activities of isocitrate dehydrogenase (ICDH) and of lactate dehydrogenase. In all four analysed muscles the GLUT4 contents of adult goats were significantly (p < 0.05) lower than in goat kids. Significant differences concerning the GLUT4 contents in skeletal muscles were not detected in adult goats of different stages of life. The GLUT1 contents differed to a lower extend between goat kids and adult goats. The results of this investigation indicate that the impaired insulin sensitivity of adult compared with suckling ruminants is accompanied by or leads to a decreased GLUT4 expression.

[Effects of excess caloric fat feeding on the lipid metabolism in Shetland ponies]. / Auswirkungen überkalorischer Fettfütterung auf den Fettstoffwechsel bei Shetland Ponys.

To investigate the influence of overweight and dietary fat supplementation on lipid and insulin glucose metabolism of Shetland ponies, eight Shetland pony geldings were fed a hypercaloric (30 MJ DE/150 kg bwt. and day) fat diet (10% fat as soybean oil) or a carbohydrate control diet for nine months until ponies gained an overweight of 15%. Afterwards oral glucose tolerance tests (oGTT; 5, 6 mmol/kg bwt.) were performed after a 12 hour fast and after a fast which led to an increase of plasma triglyceride concentrations to a threshold of 3 mmol/l (36-65 hrs.). Plasma concentrations of glucose, insulin, triglycerides and non esterified fatty acids (NEFA) were determined for 480 minutes after the glucose load. Ponys having had received the control diet tended to a higher insulin secretion in case of both oGTTs, whereas the glucose tolerance was similar in both groups but lower than in ponies of normal weight. During the oGTTs after fasting leading to the plasma triglyceride threshold, triglyceride concentrations decreased significantly (p < 0.05) faster and stronger in fat fed ponies. Additionally, fat fed pony showed significantly (p < 0.05) lower NEFA levels. The results of this study demonstrate a positive effect of fat feeding on the triglyceride clearance of overweight Shetland ponies.

Effect of dietary vitamin E supplementation on the shelf life of cured pork sausage.

This study examined the shelf life of cured sausage under different packaging conditions from vitamin E supplemented pigs. One group (n=6) of crossbred pigs received a normal fattening diet containing 20 ppm α-tocopherol for 39 days before slaughter. Another group was fed a diet containing 410 ppm α-tocopherol during the same period. After slaughter, cured sausages were produced, packaged under three different atmospheric conditions, ripened for 4 weeks and then stored for 8 weeks (9 °C; 200 lux). The α-tocopherol content was recorded in the fat, liver, muscles and the sausage. TBARS, L*, a*, b* values and antioxidative capacity were evaluated in the sausage during storage. The results showed a transfer of vitamin E into tissues and sausage but no detectable effect on TBARS and colour stability. However, antioxidative effects of vitamin E were seen by provocation. Probably the effect of vitamin E in the sausage was masked by nitrite in the curing salt. The oxygen content of the packs had an influence on TBARS and colour stability.

Influence of high levels of vitamin E on semen parameters of cocks.

This study was an attempt to find whether the reproductive performance of cocks would be influenced by oral administration of different amounts of dietary vitamin E over a long period of time. For that purpose 60 cocks were divided into five dietary groups of 12 animals each, and supplemented with 0 (control group), 100, 1,000, 10,000 or 20,000 IU alpha-tocopherol/kg diet, respectively, over a period of 12 months. The effect on semen parameters and biochemical parameters measured in pooled semen samples and the weight of the testes were determined. The weight of testes decreased with increasing amounts of supplemented vitamin E. Volume, pH, colour, consistency and motility were not influenced by the diets, but density of ejaculate (sperm/microl), total amount of spermatozoa and morphology of sperm were significantly lowered by increasing amounts of supplemented vitamin E. The alpha-tocopherol concentration in ejaculates increased significantly in relation to the diet whereas phospholipid content and thiobarbituric acid-reactive substances (TBARS) of the semen samples decreased significantly with increasing vitamin E supplementation. The reproductive performance of cocks was negatively influenced by high doses of vitamin E although decreased TBARS indicated rising oxidative defence.

Insulin responsiveness of sheep, ponies, miniature pigs and camels: results of hyperinsulinemic clamps using porcine insulin.

It had been suggested that marked species differences in glucose tolerance tests were due to differences in insulin resistance. To compare insulin responsiveness, euglycemic hyperinsulinemic clamps were carried out in sheep, ponies, miniature pigs and camels. Porcine insulin was infused as primed-continuous infusions for 2 h (6 mU x kg(-1) x min(-1)). The steady state glucose infusion rates in the pigs, sheep, ponies and camels were 96.0, 18.6, 7.1 and 6.1 micromol x kg(-1) x min(-1), respectively. The maximal plasma insulin concentrations during the insulin infusions were 2,700 microU x ml(-1) in the camels, 1,400 microU x ml(-1) in the sheep and ponies and 600 microU x ml(-1) in the pigs. The rate of insulin removal from plasma was lowest in the camels as compared to the sheep, ponies and pigs (0.019, 0.038, 0.035 and 0.070 min(-1), respectively). In all species the concentrations of plasma non-esterified fatty acids dropped significantly 10-30 min after the start of the insulin infusion. However, the rates of non-esterified fatty acid reduction were higher in the pigs and sheep than in the camels and ponies. Results confirm a considerably higher insulin responsiveness in the pigs as compared to the sheep. The ponies and camels were found to be even more insulin-resistant than the sheep.

Effects of fat feeding and energy level on plasma metabolites and hormones in Shetland ponies.

The aim of this study was to investigate the influence of a fat-supplemented diet compared with a carbohydrate diet on the lipid metabolism and the enteroinsular axis of Shetland ponies. The 'crossover' experiment was divided into two parts: in the first 10 weeks the diets comprised the correct number of calories according to requirements and in the following 10 weeks they were hypercaloric, in order to check the effect of a different energy content of the diets. Feeding the fat-enriched diet, independently of its energy content, led to a significant decrease in plasma triglycerides, associated with a mean 50% increase of plasma lipoprotein lipase activity. After oral glucose load the ponies on fat-enriched diets showed higher plasma glucose concentrations. Oral glucose administration after feeding the hypercaloric fat-enriched diet led to a 25-fold increase of plasma insulin levels. Glucose-dependent insulinotropic polypeptide plasma levels were increased in the animals on the fat-enriched diets. The results of this study suggest that fat feeding improves triglyceride clearance. However, the fat supplementation of the diet also led to impaired glucose tolerance. These results are important for a better understanding of the function of the enteroinsular axis. To investigate the influences of fat on lipid metabolism in relation to the aetiopathogenesis of equine hyperlipaemia further studies involving diseased animals are needed.

Effects of feeding high dosages of vitamin E to laying hens on thyroid hormone concentrations of hatching chicks.

Long-term experimental feeding of 20,000 ppm alpha-tocopheryl acetate to laying hens caused a significant (P < 0.05) decrease in hatching rates as compared to the control group, which was fed a diet containing 19 ppm alpha-tocopherol. When the thyroid hormones in the developing chicks were checked on incubation days 16, 19, 21, and 22, the following results were ascertained: During the latter part of incubation, increases in plasma concentrations of thyroxine and triiodothyronine were observed. No significant differences in hormone concentrations (P > 0.05) between the control and the treatment group were observed during incubation days 16, 19, and 22. However, on the day of hatching (day 21 of incubation) significantly lower (P < 0.05) triiodothyronine concentrations in chick embryos of piped eggs were found in the treatment group. Moreover, thyroxine concentrations in non-piped eggs and in hatched chicks were found to be significantly higher as compared to the control group. Given these results, one concludes that extremely high dosages of vitamin E may affect thyroid hormone concentrations of hatching chicks, and therefore, the chicks might be inhibited in pipping the egg shell. Hypothetically, the hepatic enzyme 5'-monodeiodinase is involved in the mechanism of inhibition.

Glucose-dependent insulinotropic polypeptide (GIP) and the enteroinsular axis in equines (Equus caballus).

To investigate the enteroinsular axis (EIA) in equines oral (oGTT) and intravenous (i.v.GTT) glucose tolerance tests (5.6 and 1 mmol glucose/kg BW, respectively) were performed with healthy, normal weight large horses and Shetland ponies. Plasma was analysed for concentrations of glucose, glucose-dependent insulinotropic polypeptide (GIP) and insulin. In all equines plasma GIP concentrations only increased significantly when glucose was administered orally. The insulin glucose ratio (IGR) was significantly higher during the oGTT than during the i.v.GTT in both races. Basal plasma glucose levels were significantly higher in large horses than in ponies in both experiments. During the oGTT maximum glucose values were significantly higher in ponies. Ponies tended to a higher insulin secretion but the IGRs were identical in both races after oral and intravenous glucose administration. One clinically inconspicuous pony showed hyperinsulinaemia and, in case of the oGTT, insulin resistance, glucose intolerance, and GIP hypersecretion. The results of this study indicate the existence of an EIA in equines due to the higher IGRs during the oGTT. Furthermore, the similarity of plasma GIP levels and IGRs in ponies and large horses suggest a comparable activity of the EIA in both races. Regarding the elevated plasma GIP concentrations of the insulin resistant pony the EIA appears to participate in equine hyperinsulinaemia.

Brain, liver and plasma unsaturated aldehydes in nutritional encephalomalacia of chicks.

Vitamin E deficiency and linoleic acid-feeding lead to nutritional encephalomalacia (NE) in chicks, affecting the cerebellum exclusively. The relevance of lipid peroxidation (LPO) products to the pathogenesis of the disease was studied. Laying hens received a diet low in vitamin E. Resulting chicks were assigned to four groups fed either with linoleic (C18: 2n-6) or linolenic (C18: 3n-3) acid together with 1 or 50 p.p.m. vitamin E. Nine days post-hatching NE occurred in the vitamin E-deficient group fed linoleic acid. With each chick showing NE, a healthy one from all four groups was killed. Unsaturated aldehydes were determined in plasma, liver, cerebrum and cerebellum. Results underlined that the type of dietary fat is decisive for the aldehyde pattern. In the liver of linoleic acid-fed animals total aldehydes were increased. Diseased animals had increased aldehydes stemming from n-3 fatty acids. In plasma, vitamin E deficiency led to higher malondialdehyde and OH-nonenal concentrations. In brain, neither vitamin E deficiency nor NE were accompanied by increased aldehyde concentrations. In consequence a direct role of unsaturated aldehydes for the development of NE in the cerebellum is not probable.

[New fundamental order of the School of Veterinary Medicine Hanover]. / Neue Grundordnung für die Tierärztliche Hochschule Hannover.

On June 29, 2000, the Council of the School of Veterinary Medicine Hannover (TiHo) passed a new fundamental order. Upon termination of the current terms of office of the rector and the chancellor in September and at the end of the year 2001, respectively, a school management will be established with a longer term of office and a new type of School Council. However, the new fundamental order must still be approved by the Lower Saxony Ministry of Science and Culture. This change has been made necessary by radical restructuring affecting all universities in Lower Saxony: the transformation to a state enterprise with the concomitant introduction of a global budget demands full-time management. This is a step of historical significance, for this change in the fundamental order puts an end to a tradition of 87 years at the School of Veterinary Medicine Hannover under a constitutional rectorate. The following essay describes the most important changes and explains their necessity.

A study of lipid peroxidation and vitamin E in dairy cows with hepatic insufficiency.

Sixteen dairy cows were studied to assess the status of the natural antioxidant vitamin E and lipid peroxidation in their livers. Cows with liver failure (n = 7) showed clinical signs of a hepatic encephalopathy and had the following values of selected blood indices: AST > 80 U/l and GLDH > 15 U/l in serum, and venous plasma ammonia > 35 mmol/l. The control group (n = 9) consisted of dairy cows which were recovering from surgery (omentopexy) and were free of any health complications. Blood was analysed for alpha-tocopherol, aspartate aminotransferase, glutamate dehydrogenase, gamma-glutamyl transferase, total bilirubin, ammonia, cholesterol, albumin, free fatty acids, glucose, and beta-hydroxybutyrate. Alpha-tocopherol, triglyceride and malondialdehyde were measured in wet liver tissue. The cows with hepatic failure were clearly low in alpha-tocopherol and had significantly lower (P < 0.01) plasma alpha-tocopherol than the controls. Both liver triglycerides and MDA were higher (P < 0.05) in the cows with fatty livers. It is concluded that the cows with liver failure had an increase in the intensity of hepatic lipoperoxidative processes and a low antioxidative status, which should be taken into consideration in cases where treatment of the disease is proposed.

The determination of TBA-reactive substances and alkenals in the presence of antioxidants.

This study investigated the inhibitory efficiency of all-rac-alpha-tocopherol, 2,6-ditert-butyl-p-cresole (BHT), and 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carbonic acid (Trolox) on determination of thiobarbituric acid-reactive substances (TBARS) and short-chain alkenals in rat liver homogenates. The concentration of TBARS was measured fluorophotometrically. Aldehydes were determined after derivatization with methylhydrazine by gas chromatography (GLC). The concentrations of alkenals and TBARS in liver homogenates were diminished when antioxidants were present during the sample preparation. It is suggested that in the absence of antioxidants the samples are autoxidized further during the preparative procedures. For the aldehyde determination all-rac-alpha-tocopherol was the most effective antioxidant to reduce the bias due to autoxidation, whereas for TBARS it was Trolox.

Studies on equine lipid metabolism. 2. Lipolytic activities of plasma and tissue lipases in large horses and ponies.

The enzymatic fundamentals of lipid metabolism of equine have not been thoroughly investigated at this point in time. It is still unclear why ponies in contrast to horses may become hyperlipaemic when coming negative energy balance. In this study, the activities of the triglyceride-cleaving key enzymes of ponies are large bred horses were investigated in order to obtain insight into the aetiology of the syndrome. The objective of the study was to measure the activities of hormone-sensitive lipase (HSL), lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) in ponies and horses in ex vivo in vitro assays. Norepinephrine (NE) stimulated pony adipocytes to release FFA in a linear fashion (4.57 +/- 2.09 nmol FFA.10(5) cells-1.min-1). This was not observed in horses. Lipolysis was significantly higher in fat cells of ponies than in horses when adenosine deaminase (ADA) and NE were added (12.71 +/- 3.12 vs. 1.96 +/- 1.22 nmol FFA.10(5) cells-1.min-1). Relative inhibition of lipolysis by the action of insulin was comparable in adipocytes of horses and ponies. However, absolute FFA release in pony fat cells was as high as the maximal NE and ADA stimulated lipolysis in horse adipocytes. Postheparin plasma lipase activities in ponies and horses did not differ between the sub-species. This finding was supported by the results obtained from measurement of LPL activity in adipose and muscle tissue showing only a tendency of increased activities in pony explants when compared to horse tissue incubations. This study further supports the hypothesis that differences in regulation of TG release from fat stores rather than clearance of TG from plasma is causative for the development of hyperlipaemia in ponies. Abbreviations used: ADA, adenosine deaminase; BW, body weight; FFA, free fatty acid; HSL, hormone-sensitive lipase; HTGL, hepatic triglyceride lipase; LPL, lipoprotein lipase; NE, norepinephrine; SDS, sodium dodecyl sulfate; TG, triglyceride; VLDL, very low density lipoprotein.

Effects of conditioning horses with lactate-guided exercise on muscle glycogen content.

The effects of 3 different conditioning programmes on muscle glycogen concentration in horses were examined. Speed of exercise was selected according to the blood lactate values for each horse derived from a standardised exercise test before beginning a conditioning programme. Six 2-year-old Haflinger stallions were assigned randomly to one of 3 conditioning programmes according to a 6 x 3 latin square design: 45 min at their individual v1.5 or v2.5 and 25 min at v4. Each conditioning programme lasted 6 weeks (21 exercise sessions), followed by 5 weeks without conditioning (resting period). All exercise was carried out on a treadmill inclined at 17%. Muscle biopsies were taken 5 times from the gluteus medius muscle at 2 cm and 6 cm depth: before the start and in the middle of the conditioning period, then at Days 2, 9 and 35 after the last exercise session. It was found that glycogen concentration was not affected by conditioning until 9 days after finishing conditioning at v1.5 and v2.5 for 45 min (P < 0.05). By this time glycogen concentration in the muscle samples taken at 6 cm depth increased by 47 and 48%, respectively, and remained elevated until the end of the resting period. It was concluded that conditioning at lower intensity and for longer duration seemed to increase glycogen stores in the muscle while faster intensity but shorter duration exercise did not. To increase the likelihood of measuring effects of conditioning programmes on muscle variables, sampling should be done at different depths of a muscle and at several days after finishing a conditioning programme.