The change of sleeping and lying posture of Japanese black cows after moving into new environment.

OBJECTIVE: Environmental change is one of the stressful events in livestock production. Change in environment disturbs cow behavior and cows require several days to regain a stable behavioral pattern. Sleeping posture (SP) and lying posture (LP) have been used as indicators for animal that are relaxed and well-acclimated to their environment. The aim of this study was to examine the time required by Japanese black cows for stabilization of SP and LP after moving into new environment. METHODS: Seven pregnant Japanese black cows were used. Cows were moved into new tie-stall shed and their sleeping and lying posture measured 17 times during 35 experimental days. Both SP and LP were detected by accelerometer fixed on middle occipital and hip-cross, respectively. Daily total time, frequency, and average bout of both SP and LP were calculated. RESULTS: Daily SP time was the shortest on day 1 and increased to the highest on day 3. It then decreased until day 9, after that stabilized about 65 min/d till the end of experiment. Daily LP time changed in same manner as daily SP time. The average SP bout was the longest on day 1, and then decreased to stable level on day 7. On the other hand, the average LP bout was the shortest on day 1, and it increased to stable level on day 7. CONCLUSION: These results showed that pregnant Japanese black cows needed 1 week to stabilize their SP. However, there were different change patterns between the average SP and LP bout, even though the change pattern of daily SP and LP time were similar.

Possible actions of corticotropin-releasing hormone in regulating porcine corpus luteum function.

The objective of the present study was to determine whether corticotropin-releasing hormone (CRH) influences porcine corpus luteum function. The gene expressions of CRH receptors (CRH-R) were determined in the CLs of Chinese Meishan pigs during the estrous cycle. The effects of CRH on progesterone (P), estradiol-17ß (E) and prostaglandin (PG) F(2α) secretion by cultured luteal cells were also investigated. Messenger RNAs of the CRH-R were clearly expressed in the CL throughout the estrous cycle, and the mRNA level was higher at the regressed stage than at the other stages (P<0.05). When the cultured luteal cells obtained from the mid-luteal stage CL (days 8-11) were exposed to CRH (50-5000 ng/ml), P secretion by the cells was significantly reduced at the highest dose (P<0.05), whereas CRH had no effect on E and PGF(2α) secretion by the cells. The overall results suggest that CRH inhibits local P production from luteal cells via its specific CRH-R, implying that CRH plays some roles in regulating porcine CL function during the estrous cycle, especially in the period of luteolysis.

The influence of concentrate-rich diet on daily sleep and lying postures of Japanese black cows.

Sleep is one of the essential behaviors in mammalian health and welfare. In this study, we focused on how concentrate-rich diets (Conc) can influence the sleep of cattle. We hypothesized that Conc would suppress the daily sleep of cows because of behavioral frustration according to shorter eating and rumination. We subjected six Japanese black cows according to crossover experimental design. There were two diet treatments. In the Conc group, 70% of required TDN was supplied by concentrate and 30% by grass silage. On the other hand, in roughage fed group (Rough), all required TDN was supplied by grass silage. Maintenance behaviors, postures of cows, and sleep posture were measured by electronic devices. Cows fed with Conc spent less time in eating and rumination and spent more time in rest than Rough. However, contrary to our hypothesis, cows fed with Conc showed longer daily sleep posture duration (96.2 min/day vs. 69.1 min/day) and more frequent sleep posture bouts (15.4 times/day vs. 11.1 times/day) than Rough. We discussed about following possible explanations why cows fed with Conc slept longer as satisfaction, illness, displacement behavior, and shifting sleep posture.

Production of 3-hydroxypropionaldehyde in silage inoculated with Lactobacillus coryniformis plus glycerol.

We examined the production of an antimicrobial component, 3-hydroxypropionaldehyde (3-HPA), in laboratory-scale silage inoculated with Lactobacillus coryniformis strain 394, which ferments glycerol to 3-HPA. A modified colorimetric method that used an NaOH-treated blank and determined the absorption spectrum of the samples was employed to detect a 3-HPA-like component (HLC) that was assumed to be 3-HPA. Inoculation with Lb. coryniformis 394 plus glycerol in ensiling produced HLC at 10-460 ppm and contributed to inhibition of butyric fermentation and retardation of aerobic spoilage. HLC was considered to be 3-HPA from its absorption spectrum. These results suggest that the production of 3-HPA by Lb. coryniformis 394 is useful in ensiling and that the modified colorimetric method is effective to detect 3-HPA in silage.

Sleep and lying behavior of milking Holstein cows at commercial tie-stall dairy farms.

The aim of this study was to clarify cow's sleep behavior on farm and to examine the influence of some environment and management factors on cow's sleep. Total 85 behavior data (summer 41, autumn 44) were corrected from 12 commercial tie-stall dairy farm in northern Tohoku area. Daily duration, bout frequency, and averaged bout duration of sleep and lying were measured. Even though there was no significant difference in any lying measurements, daily duration, bout frequency, and averaged bout duration of sleep was significantly different among farms (p < 0.01, p < 0.01, and p < 0.05, respectively). However, we could not find that any managements influence on sleep. There was significant seasonal difference in daily sleep duration (p < 0.01) and sleep bout frequency (p < 0.05). On the other hand, daily lying duration and averaged lying bout duration were significantly different between seasons (p < 0.001 and p < 0.05, respectively). Primiparous cows slept longer and more frequent than multiparous (both p < 0.001). Milk solid-non-fat content was positively correlated with daily sleep duration (p < 0.05) and bout frequency (p < 0.01). These results might imply the productive importance of sleep related with anabolic hormones release.

The use of accelerometer to measure sleeping posture of beef cows.

Sleep is one of the essential behaviors for mammals. The aims of this study were to validate the use of accelerometer for measuring sleeping posture of cattle. Duration of sleeping posture of seven Japanese Black cows from 19.00 to 07.00 hours was measured by both accelerometer and video, and a total of 67 accelerometer and video measurement sets were collected. We calculated Cohen's κ coefficient between accelerometer and video measurements and 91.5% of the κ-values were >0.80. Intra- and inter-observer coefficient of variance showed that specific acceleration waveform patterns of sleeping posture could be easily and accurately detected by independent observers. There were no significant differences in the frequency of sleeping posture occurrences between accelerometer and video measurements. We compared averaged sleeping posture bout, and the total sleeping posture time between accelerometer and video measurements using regression. In each trait, the slope was close to 1 and the intercept was not different from 0, which showed a strong agreement between accelerometer and video measurements. This shows that an accelerometer could accurately detect sleeping postures of cattle. We conclude that adequate measurements of sleeping postures can be made using an accelerometer.

Relationship between the stockperson's attitudes and dairy productivity in Japan.

The aim of this study is to identify the factors that comprise farmer attitudes toward dairy cows and jobs in Japan, and examine the relationship between these attitudes and dairy productivity. At first, we executed a questionnaire survey to determine factors that comprise attitudes of farmers toward their jobs and dairy cows, and three factors were extracted. These were named as 'Positive beliefs to cows', 'Negative beliefs to cows' and 'Job satisfaction', respectively. Second, we examined the relationships between attitude and dairy productivity in 35 dairy farms. The positive beliefs scores correlated positively both with milk yield and milk urea nitrogen concentration. We found there to be three farm groups by cluster analysis using three attitude score. The group B farms showed significantly higher positive beliefs scores and job satisfaction scores; on the other hand, the group C farms showed significantly lower positive beliefs scores and higher negative belief scores. The milk yield in group B was significantly higher than that in group C. This study showed that Japanese farmers' attitudes toward cows considerably resemble those seen in previous studies in Western cultures. Positive attitudes toward cows could enhance stockmanship, and could improving animal welfare and productivity.

Growth hormone and lactogenic hormones can reduce the leptin mRNA expression in bovine mammary epithelial cells.

Leptin mRNA is expressed in not only adipocytes but also mammary epithelial cells and leptin protein is present in milk. Although milk leptin is thought to influence metabolism or the immune system in neonates, there is little information about the regulation of leptin expression in mammary epithelial cells. We examined the effect of growth hormone (GH) and/or lactogenic hormone complex (DIP; dexamethasone, insulin and prolactin) on leptin mRNA expression in mammary epithelial cells. We used a bovine mammary epithelial cell (BMEC) clonal line, which was established from a 26-day pregnant Holstein heifer. We confirmed that the mRNA was expressed in BMECs and the expression was significantly reduced by GH and/or DIP, when the cells were cultured on both plastic plates and cell culture inserts at days 2 and 7 after stimulation with lactogenic hormones. GH and/or DIP significantly increased level of alpha-casein mRNA in BMECs after 7 days on the cell culture inserts, but no mRNA expression was detected at day 2. GH and DIP significantly stimulated the secretion of alpha-casein from BMEC on cell culture inserts at 3.5 and 7 days. However, neither alpha-casein mRNA expression nor secretion was observed in the BMECs cultured on plastic dishes, even in the presence of GH or/and DIP. These results indicate that GH and DIP can directly reduce leptin mRNA expression in both undifferentiated and functionally differentiated bovine mammary epithelial cell.

Expression of mRNAs for interleukin-4, interleukin-6 and their receptors in porcine corpus luteum during the estrous cycle.

There is increasing evidence that inflammatory cytokines regulate corpus luteum (CL) function in many species. The purpose of the present study was to determine whether interleukin (IL)-4 and IL-6 are expressed in the porcine CL, and whether these cytokines influence porcine luteal steroidogenesis. The gene expressions of IL-4, IL-6 and their specific receptors were determined in the CL of Chinese Meishan pigs during the estrous cycle. Moreover, the effects of these cytokines on progesterone (P(4)), estradiol-17beta (E(2)) and prostaglandin (PG) F2alpha secretion by cultured luteal cells were investigated. IL-4 and IL-6 mRNAs were detected in the CL at all luteal stages. Furthermore, mRNAs of the receptors for IL-4 and IL-6 were clearly expressed in the CL throughout the estrous cycle. Real-time PCR analysis revealed that IL-6 receptor (IL-6R) mRNA expression was higher in the regressed CL (days 19-21 after ovulation) than in the CL at other stages (P<0.01). Exposure of cultured luteal cells obtained from mid-stage CL (days 8-11) to IL-6 (1-100 ng/ml), it inhibited P(4) and E(2) secretion by the cells (P<0.05). Although IL-4 (1-100 ng/ml) did not significantly alter P(4) secretion, it inhibited E(2) secretion by the cells (P<0.05). Neither IL-4 nor IL-6 had any effect on PGF2alpha secretion by the cells. These results suggest that IL-4 and IL-6 are locally produced in the porcine CL, and that they inhibit steroid production from luteal cells via their specific receptors. Collectively, both IL-4 and IL-6 may play roles in regulating porcine CL function throughout the estrous cycle.

Effects of fasting and refeeding on expression of proteolytic-related genes in skeletal muscle of chicks.

This experiment was conducted to study the effects of fasting and refeeding on proteolytic-related gene expression in skeletal muscles of chicks. Chicks were fasted for 24 h, and refed for 2 h. Plasma Ntau-methylhistidine concentration, as an index of myofibrillar protein degradation, was increased by fasting, and that increment was reduced by refeeding. We also examined the expression of the protease mRNAs (calpain, proteasome, cathepsin and caspase-3) by real-time PCR of cDNA in skeletal muscles of fasting and refeeding chicks. Calpain (m-, mu-, and p94/calpain-3) mRNA expressions were also increased by fasting, and their increment was reduced by refeeding. Ubiquitin and 20S proteasome alpha subunit (alpha6 and alpha7) mRNA expressions as well as cathepsin B, and caspase-3 mRNA expression were likewise increased by fasting, with their increment also reduced by refeeding. These results indicate that fasting stimulates proteolytic-related gene expression, resulting in an increase in myofibrillar protein degradation, and that refeeding suppresses proteolytic-related gene expression, resulting in a decrease in myofibrillar protein degradation in chicks.

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.

Physiological changes during feeding and rumination in cows.

In this study, we investigated the physiological changes in cattle during feeding and rumination. We collected blood samples every 5 min by using an automated blood sampling system and simultaneously recorded feeding, ruminating, and other behaviors using a video camera. Plasma non-esterified fatty acid (NEFA) concentrations continuously decreased during feeding and decreased temporarily during rumination. Plasma glucose concentration continuously decreased during feeding and remained stable during rumination. During feeding and rumination, there were no characteristic increases and subsequent decreases in plasma insulin and growth hormone (GH) concentrations, although insulin concentrations were positively correlated with glucose concentration. NEFA concentrations were not correlated with GH and insulin concentrations. In terms of chewing behavior, feeding and rumination are similar; therefore, the changes in metabolites such as NEFA might have been the same. Combination of behavioral observations and application of an automated blood sampling system could contribute to new findings on behavioral and physiological changes regarding the temporary decrease in plasma NEFA concentration during rumination in ruminants.

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.

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.

Growth hormone acts on the synthesis and secretion of alpha-casein in bovine mammary epithelial cells.

To study the effect of growth hormone (GH) on the functions of mammary epithelia, we examined the effect of GH on the synthesis and secretion of alpha-casein in a bovine mammary epithelial cell (BMEC) clonal line, which was established from a 26-d-pregnant Holstein heifer. GH receptors (GHR) were observed in the BMEC on the membrane as well as in the cytoplasm. After BMEC were plated onto cell culture inserts, GH stimulated alpha-casein release in both the presence and absence of the lactogenic hormone complex, which included dexamethasone, insulin and prolactin (DIP). DIP enhanced the effect of GH on alpha-casein release. Although alpha(s1-) casein mRNA expression was not detected in untreated control cells, its expression was observed in BMEC in response to the GH, DIP and GH + DIP treatments. Expression was greater for GH and GH + DIP than for just DIP. Expression of GHR mRNA was increased by DIP treatment, while the mRNA expression was little changed by GH treatment. We conclude that GH acts on BMEC and induces the expression of both the alpha-casein gene and protein. GHR gene expression was shown to be regulated by DIP and GHR. GHR may be involved in a synergic effect between GH and DIP on casein secretion. These results suggest that GH, in addition to its widely accepted homeorhetic role in vivo, also can act on the mammary parenchyma, and that the effects of GH on mammary epithelial cells could partly account for the clear galactopoietic effect of recombinant bovine GH seen in lactating dairy cows.

In vitro differentiation of a cloned bovine mammary epithelial cell.

The aim of the study was to establish in vitro a bovine mammary epithelial cell (MEC) clone, able to respond to mitogenic growth factors and to lactogenic hormones. Mammary tissue from a 200-d pregnant Holstein cow was used as a source of MEC, from which a clone was established through a process of limiting dilution. When plated on plastic, the cells assumed a monolayer, cobblestone, epithelial-like morphology, with close contact between cells. Inclusion of IGF-1 and EGF in the media significantly increased the number of cells 5 d after plating. All cells stained strongly for cytokeratin and moderately for vimentin at young and old passage stages, indicating the epithelial nature of this cell clone. When the cells were plated at a high density on a thin layer of a commercial extracellular matrix preparation (Matrigel), lobular, alveoli-like structures developed within approximately 5 d, with a clearly visible lumen. When cells were plated onto Matrigel in differentiation media (containing lactogenic hormones), detectable quantities of alpha-casein were present in the media and particularly on the lumen side of the structures. Omission of one of the lactogenic hormones (insulin, prolactin or hydrocortisone) reduced alpha-casein release to the limit of detection of the assay used. Lactoferrin was also produced when the cells were plated on Matrigel, again principally on the lumen side of the lobules, though this was independent of the lactogenic hormones. By passage 40, the cells had senesced, and it was not possible to induce alpha-casein or lactoferrin production. This study notes the establishment of a functional bovine mammary epithelial cell clone, which is responsive to mitogenic and lactogenic hormones and an extracellular matrix.