Maximum permissible load for Kiso horses trotting over a short, straight course.

This study aimed to determine the load-bearing capacity of trotting Kiso horses using gait analysis. Ten Kiso horses with a height at withers of 128 cm were included. Their riders were fitted with a marker (70 mm in diameter placed on their chest) recorded by two digital DVD cameras while trotting along a short, straight course. In total, eight tests were performed for each horse: the first with a 70 kg load; six with randomly loaded weights ranging 80-130 kg; and then a final test again with a 70 kg load. Three-dimensional movement of the marker was analyzed using a motion capture system. The time series of vertical displacement of the marker underwent spectrum analysis by the maximum entropy method, and the autocorrelation coefficient was calculated. The first two peaks of the autocorrelation were defined as symmetry and regularity, and their sum was defined as stability. Regularity in the 120 kg test (0.54) was lower than that in the first 70 kg test (0.61), and stability in the 120 kg test (1.31) was lower than that in the first 70 kg test (1.42). We concluded that the maximum permissible load for a trotting Kiso horse is < 120 kg, which represents 31% of its bodyweight.

The beneficial effects of horse trekking on autonomic nervous activity in experienced rider with no disability.

The aim of this study was to determine whether autonomic nervous activity of a rider with no disability was altered by one practical and applicable horse trekking (HT) exercise. Changes in autonomic nervous activity were analyzed by heart rate variability (HRV). Twenty-three participants with no disability rode horses along a predetermined HT course at trotting and walking for 60 min. HRV was sampled at 60 min before and immediately, following 60 min, and 120 min after HT. As a control, the same measurements were performed for 22 age-matched participants during their rest. Only in the HT group, the value of normalized unit in high frequency component (HF nu), an index of parasympathetic nervous activity, was higher at 120 min after treatment than before HT (P < 0.05). The low / high frequency ratio (LF / HF), believed to reflect sympathetic nervous activity, was lower in the HT group than those in the control group at 60 min (P < 0.05) and 120 min after treatment (P < 0.01). These findings suggest that a single HT shifted the autonomic nervous balance of a rider toward parasympathetic dominance. The results obtained by the present study could accelerate the use of horses for human health.

Influence of riders' skill on plasma cortisol levels of horses walking on forest and field trekking courses.

The aim of this study was to evaluate the influence of rider's skill on the plasma cortisol levels of trekking horses on two courses, walking on field and forest courses (about 4.5 to 5.1 km each). Three riders of different skills did horse trekking (HT) in a tandem line under a fixed order: advanced-leading, beginner-second and intermediate-last. A total of six horses were used and they experienced all positions in both courses; a total of 12 experiments were done. Blood samples were obtained before HT, immediately after and 2 h after HT. As a control, additional blood samples were obtained from the same horses on non-riding days. Irrespective of the course and the rider's skill, the cortisol level before HT was higher than that of control (P < 0.05). In both courses, the cortisol levels immediately after HT ridden by the advanced rider were higher than that of control (P < 0.05). However, in every case, the cortisol level 2 h after HT was closely similar to the level of the control. Thus, we concluded the stress of trekking horse was not sufficient to disturb the circadian rhythm of the cortisol level, irrespective of the course and the rider's skill.

Effect of melatonin injected into the third ventricle on growth hormone secretion in Holstein steers.

The effects of melatonin (MEL) injection into the third ventricle (3V) on growth hormone (GH) secretion were investigated in conscious Holstein steers. A stainless steel cannula was stereotaxically implanted in the 3V based on the ventriculogram. In Exp. 1, three doses of MEL (100, 300 or 600 microg) were injected into the 3V through the cannula and the GH concentration after the injection was determined. In Exp. 2, intracerebroventricular (icv) and intravenous (iv) injections of MEL (100 microg) and GH-releasing hormone (GHRH; 0.25 microg/kg body weight), respectively, were performed simultaneously to examine the effect of MEL on GHRH-induced GH release. The icv injection of MEL significantly stimulated GH release at 100 microg. The increase in GH concentrations by 100 microg of MEL was persistent. Intravenous injection of GHRH dramatically increased GH release. The injection of MEL did not alter GHRH-induced GH release. These results suggest that MEL stimulates GH secretion possibly through the hypothalamus in cattle.

A novel stereotaxic approach to the hypothalamus for the use of push-pull perfusion cannula in Holstein calves.

To determine secretory patterns of growth hormone-releasing hormone (GHRH) and somatostatin (SS) and their roles in the regulation of growth hormone (GH) secretion, a method for collecting hypothalamic perfusates, a push-pull perfusion method was developed in calves. With the use of the stereotaxic apparatus for cattle, a cannula was implanted into the hypothalamus of four male calves based upon cerebral ventriculography. Push-pull perfusates were collected at 10 min intervals for 6h and GHRH and SS concentrations in perfusates and plasma GH concentration were determined by EIAs and RIA, respectively. A cannula was implanted into the hypothalamus based on the image of the third ventricle and maintained for 1 month. GHRH and SS showed pulsatile secretion and the pulses for GHRH and SS were irregular in conscious animals. Neither GHRH nor SS secretion had a clear relationship with GH secretion. In the present study, we thus (1) established a stereotaxic technique for approaching the hypothalamus using cerebral ventriculography for calves, and (2) demonstrated that GHRH and SS secretion were pulsatile but not closely related to GH profile in conscious calves. The technique is useful for the study of the functions of the hypothalamus in the control of pituitary hormones in cattle.

Gene expression of resistin in adipose tissue and mammary gland of lactating and non-lactating cows.

Resistin has been suggested to induce insulin resistance in obesity and to inhibit adipocyte differentiation. In lactating cows, glucose uptake in the mammary gland is a rate-limiting step in milk synthesis, and to supply glucose to the mammary gland, insulin resistance increases. We examined the expression of the resistin gene by real-time PCR of cDNA in the adipose tIssue and mammary gland of lactating and non-lactating cows. Lactation induced a significant increase of resistin expression in adipose tIssue compared with that in the dry period, and decreased resistin expression in the mammary gland. There were no significant differences in the expression of insulin responsive glucose transporter (GLUT4) mRNA between the adipose tIssue of lactating and non-lactating cows, and GLUT4 mRNA was not detected in the mammary gland. The plasma insulin concentration was lower in lactating cows than in non-lactating cows. These results indicate that the pattern of resistin expression in peripheral tIssues is changed in association with milk production. The increase of resistin expression and maintenance of a lower level of plasma insulin concentration may decrease glucose availability by increasing insulin resistance in adipose tIssue. Additionally, our results suggest that the decrease of resistin expression in the mammary gland may influence on the insulin-dependent glucose uptake in mammary epithelial cells during lactation.

Plasma cortisol and prolactin secretion rhythms in cattle under varying external environments and management techniques.

The secretion rhythms of plasma cortisol (CORT) and prolactin (PRL), hormones related to stress responsiveness and biological rhythm and controlled by light and temperature, were investigated under varying external environments and different management techniques. Serial blood samples were collected from female cattle reared in free-stall and freely fed (FF) conditions (n = 4) or in tie-stall and restricted feeding (RF) conditions (hay and concentrate twice daily, n = 4). Plasma CORT and PRL concentrations, eating behavior, and environmental parameters were analyzed. Cyclic patterns for each parameter were examined using spectral analysis, and correlations between CORT, PRL and other parameters were investigated using cross-spectral analysis. Under FF conditions, CORT secretion was not related to the lighting intensity and eating behavior. However, under RF conditions, the CORT secretion rhythm showed a distinct correlation with lighting intensity and eating behavior. Under FF conditions, the PRL secretion rhythm was similar in all seasons. However, under RF conditions, the PRL rhythm oscillated with high frequency in summer and low frequency in winter, indicating a seasonal change in rhythm. The present study demonstrates that hormone secretion rhythms change under different environments and management techniques.

Method for estimating maximum permissible load weight for Japanese native horses using accelerometer-based gait analysis.

The aim of this study was to establish a method for estimating loading capacity for Japanese native horses by gait analysis using an accelerometer. Six mares of Japanese native horses were used. The acceleration of each horse was recorded during walking and trotting along a straight course at a sampling frequency of 200 Hz. Each horse performed 12 tests: one test with a loaded weight of 80 kg (First 80 kg) followed by 10 tests with random loaded weights between 85 kg and 130 kg and a final test with a loaded weight of 80 kg again. The time series of acceleration was subjected to fast Fourier transformation, and the autocorrelation coefficient was calculated. The first two peaks of the autocorrelation were defined as symmetry and regularity of the gait. At trot, symmetries in the 100, 110, and 125 kg tests were significantly lower than that in First 80 kg (P < 0.05, by analysis of covariance and Sidak's test). These results imply that the maximum permissible load weight is less than 100 kg, which is 29% of the body weight of Japanese native horses. Our method is a widely applicable and welfare-friendly method for estimating maximum permissible load weights of horses.

Biological rhythms related to metabolism in Japanese Shorthorn cattle under varying environments and management techniques.

Plasma insulin (INS), thyroxin (T4 ), glucose (GLU), non-esterified fatty acid (NEFA), blood urea nitrogen (BUN), rectal temperature (RT) and eating behavior were evaluated in Japanese Shorthorn cattle under varying external environments and management techniques. Serial blood collection and assessments of RT and eating behavior were performed over 48 h in the spring, summer, autumn and winter in four female cattle reared under either free-stall and ad libitum feeding (FA) conditions or tie-stall and restricted feeding (TR) conditions. Cycle patterns for each parameter were examined using spectral analysis, and correlations between parameters were investigated using cross-spectral analysis. Rhythms for all parameters, except eating behavior and T4 , did not differ significantly among the varied external environments and between management techniques, although seasonal differences in the concentration or value of parameters were observed. An approximate 3- or 4-h rhythm cycle detected in T4 , GLU, NEFA, BUN, and RT might be the common metabolic rhythm. Under both conditions, the metabolite levels showed strong correlations with eating behavior. Moreover, GLU positively correlated with INS at lag time of 0 h, as did eating behavior and RT.

Plasma melatonin secretion rhythms in cattle under varying seasonal conditions.

The purpose of the present study was to investigate the secretion cycles of melatonin (MEL) in cattle over the course of four seasons. Four female Japanese Shorthorn cattle under free-stall and ad libitum feeding conditions were used, and plasma MEL concentrations were measured over a 48 h period at 1 h intervals. The time-series data were analyzed by spectral analysis, and the cycle hour was determined. Data indicated that the secretion cycle for MEL was approximately 23.5 h for all four seasons. The area under the curve of MEL from start to end of experiment for 48 h did not differ significantly among the four seasons. However, the duration of high MEL secretion which defined the duration time of the values were more than 10 pg/mL and differed significantly among the four seasons. In conclusion, this study, which was the first to use spectral analysis to evaluate the cyclic rhythm of MEL in cattle, revealed that MEL secretion cycles did not differ among the seasons. These findings are inconsistent with previous study results in that previous reports suggested that the MEL secretion cycle differed under different lighting conditions.

The effect of lighting conditions on the rhythmicity of growth hormone secretion in Holstein steers.

Growth hormone (GH) secretion regularity and the effects of lighting condition and GH-releasing hormone (GHRH) on GH release were determined in steers. First, steers were kept under 12:12 L : D conditions (light: 06.00-18.00 hours). The animals were then subjected to a 1-h advancement in lighting on/off conditions (05.00 and 17.00 hours, respectively). Blood was sampled for 24 h at 1-h interval on the seventh day of each condition. Second, GHRH was injected intravenously (IV) at 12.00 and 00.00 hours under 12:12 L : D and blood was sampled at 15-min interval for 4-h (1 h before and 3 h after the injection). Plasma GH concentrations were measured by a radioimmunoassay. Periodicity of GH secretory profile was calculated by power spectrum analysis using the maximum entropy method. Plasma GH concentrations showed a characteristic pattern consisting of four distinct peaks. Mean periodicity of GH secretory profile was 5.7 h, and it was not altered by any change in lighting conditions. IV injection of GHRH increased GH secretion during the day and night. The increase in GH secretory volume after GHRH injection during the night was equal to that during the day. The present results suggest that GH secreted from the anterior pituitary have regularity in steers.

Heart rate variability after horse trekking in leading and following horses.

Horse trekking (HT) is having a stroll on a horse along a walking trail in a forest, field, and/or sandy beach. Generally in HT, horses exercise in tandem line outside the riding facilities. Because the leading horse will be confronted with stressors in the forefront, we hypothesized that the leading horse shows higher stress responses than the following one. In order to verify the hypothesis, we compared short-term stress responses between each position in six horses. Exercise consisted of 15 min of ground riding and 45 min of HT with walking and trotting. Heart rate variability was analyzed for 5 min at 30, 60, and 90 min after the exercising period. There was no significant difference in heart rate during exercise between leading and following positions. The high frequency / low frequency power band of heart rate variability, an index of sympathetic nervous activity, after exercise, tended to be higher in the leading position than following one (P < 0.1). The result in this study can suggest that the leading horse was in a higher stressed state than the following horse after HT.

Influence of orally administered bovine lactoferrin on lipid metabolism in lipopolysaccharide-injected preruminant calves.

ABSTRACT The aim of this study was to investigate the influence of oral lactoferrin (LF) administration on lipid metabolism changes in calves given lipopolysaccharide (LPS). Twenty-one 4-day-old Holstein calves were divided into three groups, with each group receiving one of three oral doses of LF (0, 1, 3 g/day) for 10 consecutive days (day -10 to day -1). All calves were intravenously injected with LPS (50 ng/kg BW) on day 0, the day after LF treatment ended. Plasma triglyceride concentrations were lower (P < 0.05) in the LF-treated calves than in the control calves given 0 g/day of LF at 12 and 24 h after LPS injection. Plasma NEFA concentrations were elevated between 6 and 24 h after LPS treatment. At 12 h, the concentration of plasma NEFA was lower (P < 0.05) in the calves given LF 3 g/day than in the control calves. On day 0, plasma total cholesterol and phospholipid concentrations tended to be lower in the LF groups administered 1 and 3 g of LF/day than in the control group, but did not differ significantly among the groups. The plasma very-low-density and low-density lipoprotein concentrations were lower (P < 0.05) at 12, 24, and 72 h in the LF groups than in the control calves. The concentrations of plasma high-density lipoprotein tended to be lower in the LF groups than in the control group between day 0 and 96 h, though there were no significant group differences. The concentration of plasma interleukin-1beta was lower (P < 0.05) in the calves fed LF 3 g/day than in the control calves at 2 and 12-48 h after LPS injection. These data suggest that LF inhibits LPS-induced alterations in lipid metabolism in preruminant calves.

Effects of ghrelin injection on plasma concentrations of glucose, pancreatic hormones and cortisol in Holstein dairy cattle.

Ghrelin affects not only growth hormone secretion but also nutrient utilization and metabolic hormone secretion in humans and experimental animals. The effects of ghrelin on plasma metabolic hormone and metabolite levels in domestic herbivores remain unclear despite the fact that the physiological characteristics of nutrient digestion and absorption imply specific responses to ghrelin. Therefore, the effects of ghrelin on plasma glucose, pancreatic hormones and cortisol concentrations were investigated in Holstein dairy cattle in various physiological states. Ghrelin (0.3 nmol/kg) or placebo (2% bovine serum albumin in saline) was intravenously injected in pre-ruminant calves (pre-rumen function), adult non-lactating (functional rumen) and lactating cows (functional rumen and lactation), and plasma glucose, insulin, glucagon and cortisol concentrations were then determined. Ghrelin injection increased plasma glucose concentrations in adult cows, especially in lactating cows. No hyperglycemic response was observed in pre-ruminant calves. A transient rise of insulin and glucagon levels was distinctively found in lactating cows in response to the ghrelin administration. Ghrelin injection decreased the insulin level in pre-ruminant calves. Ghrelin increased cortisol secretion independently of the physiological state. The results of the present study suggest that the effects of ghrelin on plasma glucose and pancreatic hormone levels may reflect differences in the physiological states of dairy cattle.

Breed differences in growth hormone and insulin secretion between lactating Japanese Black cows (beef type) and Holstein cows (dairy type).

This study was performed to clarify the levels of growth hormone (GH) and insulin (INS) secretions and the glucose response to INS during lactation in a representative beef breed in Japan, Japanese Black cows, and to compare them with their counterparts in a dairy breed, Holstein cows. Six Japanese Black and seven Holstein primiparous cows received a single intravenous injection of GH-releasing factor (GRF; 0.25 microg/kg), glucose (112.5 mg/kg), or INS (0.2 U/kg) from late pregnancy (2 weeks antepartum) to mid-lactation (6 months postpartum). Japanese Black cows had one-tenth of the total milk yield of Holstein cows during lactation, and significantly lower GRF-induced GH and higher glucose-induced INS secretions than Holstein cows at all stages. In Japanese Black cows, even with lactation, these secretions remained essentially unchanged, whilst Holstein cows showed higher GH and lower INS secretions after the onset of lactation as compared with cows in late pregnancy. Both breeds had similar glucose response to INS at the respective stages. These results suggest that, during lactation, Japanese Black cows may minimize the catabolic effects of GH and sustain the anabolic effects of INS, in contrast with Holstein cows, but have similar ability to inhibit INS-mediated glucose utilization in peripheral tissues to Holstein cows.