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The experiment was carried out to assess the effects of feeding apple pomace silage (APS) on blood parameters in Suffolk ewes. Three ewes were used in 3×3 Latin square design over three periods with three dietary treatments as hay diet, low ethanol APS (L-APS, ethanol 48.7 g kg-1 DM) diet and high ethanol APS (H-APS, ethanol 87.2 g kg-1 DM) diet. Alfalfa hay cube and APS provided half of 110% TDN requirement for APS diets. The area upper or under the curve (AUC) of plasma ethanol was greater (P

We measured the growth performance and meat quality of 10 crossbred (Yorkshire × Duroc × Landrace) neutered male pigs to evaluate the effects of apple pomace-mixed silage (APMS). The pigs were divided into two groups and were respectively fed the control feed and the AMPS ad libitum during the experiment. No difference was found in the finished body weight, average daily gain, carcass weight, back fat thickness or dressing ratio between the control and the AMPS treatments, but average dairy feed intake (dry matter) was significantly lower and feed efficiency was significantly higher using the APMS treatment (P

Four Japanese black beef cows were used in a 4 × 4 Latin square to evaluate the fermentation quality, digestibility, ruminal fermentation and preference of total mixed ration (TMR) silages prepared with differing proportions of apple pomace (AP). Experimental treatments were the control (no AP added, CAP), 5% (low, LAP), 10% (medium, MAP) and 20% (high, HAP) of TMR dry matter (DM) as AP. All TMR silages were well preserved. Ethanol was produced in silages containing AP and the amount increased with the proportion of AP (P

Two experiments were undertaken to investigate the feeding effect of alcoholic apple pomace silages (APSs) on performance traits, ruminal pH and blood variables in sheep. In experiment 1, four ewes were fed alfalfa hay cube and either APS (ethanol content was 33.4 g /kg DM) or concentrate (control) at a ratio of 30:70 of TDN requirement for maintenance and daily 100g gain in a 2×2 crossover design over two 35-day periods. Total body weight gain was higher in control treatment. No significant treatment effect was observed for TDN intake and ruminal pH. Plasma concentrations of ethanol, lactate and ?-hydroxybutyrate (BHBA) were increased after feeding APS. Plasma insulin, glucose, glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) were not significantly differ between the treatments. The area upper or under curves (AUCs)of plasma ethanol and BHBA were greater and lactate was tended to greater in APS treatment. In experiment 2, low and high ethanol contained APSs (L-APS, 44.1g and H-APS, 66.9 g of ethanol/kg DM) were compared in the same design (Experiment 1). No significant treatment effect was observed for body weight gain and TDN intake. Pre-feeding ruminal pH was lower in L-APS treatment. Insulin concentration was slightly higher in L-APS treatment. However, no significant treatment effect was detected for other variables. According to the consistent increase in plasma ethanol, lactate and BHBA, and the plasma AUCs response of ethanol, lactate and BHBA after feeding of alcoholic-fermented APSs, it would be concluded that APS ethanol consumption affect the post prandial blood metabolism as well as influenced the plasma lactate and BHBA concentrations in the blood.DOI: http://dx.doi.org/10.3329/bjas.v43i3.21655 Bang. J. Anim. Sci. 2014. 43 (3): 224-231

Contents Mitochondria–cytoskeleton interactions were studied in the hamster embryos during interphase and M phase of the cell cycle. Two-cell embryos were cultured for 1 h with nocodazole, cytochalasin D or in a combination of both inhibitors and then centrifuged at 10 000×g for 2 min. The control embryos were only centrifuged with no inhibitor treatment. Centrifuged embryos were fluorescently stained to examine the distribution of active mitochondria and nuclear configuration. In the control 2-cell embryos, most mitochondria were accumulated at the perinuclear region with some at the cell cortex. Neither each inhibitor nor centrifugation did affect the distribution of mitochondria in interphase blastomeres. However, mitochondria were spun down towards the centrifugal pole in 71% (n = 41) of the interphase blastomeres treated with centrifugation following a combination of nocodazole plus cytochalasin D, suggesting that both microtubules and microfilaments may involve in mitochondrial redistribution during interphase of the cell cycle. In contrast, when M-phase blastomeres were treated with all drug treatments applied, including cytochalasin D, mitochondria had been usually dislocated in a unipolar cluster, suggesting that microfilaments, not microtubules, may involve in the mitochondrial redistribution during M phase of the cell cycle. The data indicate that microfilaments function in mitochondrial redistribution regardless of the stages of the cell cycle and that microtubules may strongly associate with mitochondria during the interphase but dissociate from them during the M phase.

The digestibility, nitrogen retention and ruminal fermentation of dry rice straw (DRS), untreated rice straw silage (NRS), lactic acid bacteria (LAB) treated rice straw silage (LRS) and urea treated rice straw silage (URS) were compared in a feeding trial with wethers supplemented with concentrate. Both NRS and LRS provided good fermentation quality although the lactic acid content in LRS was higher than in NRS (P 

In the present study, we investigated the effects of a specific Rho kinase (ROCK) inhibitor, Y-27632, on germinal vesicle breakdown (GVBD), changes of cytoskeletal organization, and the resumption of porcine oocyte maturation after its removal. In Experiment 1, cumulus-oocyte complexes (COCs) cultured in medium containing Y-27632 for 24 h showed GVBD of 98, 88, 84, and 4% (P

The objective of this study was to determine the fermentation quality and nutritive value of whole crop rice silage (WCRS) treated with a monosodium glutamate fermented by-product (MGB), and its effects on the digestion of WCRS using four Japanese Black cows. The whole crop rice was chopped into 3–5 cm lengths after being harvested at the yellow-ripeness stage and ensiled for 30 days after being treated with 0 (CMG), 2.5 (LMG), 5 (MMG) and 10% (HMG) levels of MGB. Samples of whole crop rice and that of WCRS were collected and analyzed for fermentation characteristics. This was followed by a nutrient digestion study using four Japanese Black cows in a 4 × 4 Latin square design. The MGB treatment improved (P 0.05) affect N retention. The digestibility of acid detergent fiber (ADF) and NDF increased with MGB treatment, and the degree was also affected by the level of MGB. The ruminal ammonia-N at 4 h after feeding was higher in MGB treatments than in the CMG treatment. This study demonstrated that although improvement on fermentation quality and digestibility of MGB treatment were not notable; however, MGB-treated WCRS have beneficial effects if fed for a sufficiently long term.

Four Japanese Black cows were studied in a 4 × 4 Latin square to examine digestibility, unhulled rice excretion and ruminal fermentation when fed forage paddy rice silage (FPRS) prepared using combine-type (CBS), flail-type (FLS), chopper-type (CPS) and multipurpose-type (MS) harvesters. FPRS samples were prepared at the yellow-ripeness stage and were treated with Lactobacillus plantarum except MS. These diets were fed to the cows for a digestion trial after they had been ensiled for 270 days. FLS and CPS exhibited lower pH and higher lactic acid content than CBS (P < 0.05). No treatment differences were found in digestibility, total digestible nutrients, digestible crude protein and nitrogen retention. The excretion of unhulled rice in feces was highest in the FLS treatment and lowest in the CBS treatment (P < 0.05). No differences were found in pH or volatile fatty acid concentrations in ruminal fluid. The ruminal ammonia-N at 0 and 4 h after feeding tended to be lower in the MS treatment than in the other treatments.

The objective was to evaluate effects of feeding level of milk replacer on body growth, plasma metabolite and insulin concentrations, and allometric growth of visceral organs in suckling calves. Holstein bull calves (n = 8; 3-4 days of age) were fed either a low amount (average 0.63 kgDM/day, LM) or high amount (average 1.15 kgDM/day, HM) of high protein milk replacer until they were slaughtered at 6 weeks of age. Body weight (BW) at 4, 5, and 6 weeks of age, feed intake, average daily gain, and feed efficiency were higher in the HM than LM calves. The HM group had higher plasma glucose at 3 and 4 weeks of age and insulin levels after the age of 4 weeks compared with LM calves whereas no effect was detected on plasma nonesterified fatty acid or urea nitrogen concentrations. The HM calves had greater empty body weight (EBW), viscera-free BW and most of the organs dissected than LM calves. Relative weights (% of EBW) of liver, spleen, kidneys, and internal fat were higher, whereas head and large intestine was lower in HM than LM calves. The results suggest that increased milk feeding levels would accelerate the growth of the body and specific organs.

To clarify the cytological characteristics of the porcine oocytes having meiotic competence, the oocytes retrieved from small (1-2 mm in diameter) and large follicles (3-6 mm) were compared. Cumulus- lacking oocytes were also examined to distinguish the characteristics between growing and atretic follicular oocytes. Distributions of cytoskeleton and mitochondria were observed by fluorescence staining in relation to the germinal vesicle (GV) stages, classified as GV 0 and GV I - GV IV. Oocyte size (107.2 ± 0.6 µm vs. 116.9 ± 0.7 µm, p

The present study was carried out to detail the cellular localization of leptin (Lep) and the leptin receptor (LepR) in the bovine adenohypophysis. Lep immunoreactivity (Lep-ir) was found in about 30% of adenohypophysial cells in the gland. Immunochemistry of Lep and specific hormones using serial sections revealed that Lep-ir was present in 60.4% of somatotrophs, 15.9% of gonadotrophs, 6.5% of mammotrophs, 6.5% of thyrotrophs and 2.4% of corticotrophs. Both the common short isoform (OBRa) and the long isoform (OBRb) of LepR mRNA were expressed in the bovine adenohypophysis. LepR immunoreactivity (LepR-ir) was found in only 2.8% of the adenohypophysial cells and over 50% of LepR-ir cells were gonadotrophs, in which most of the cells were distributed in the zona tuberalis. The findings on Lep and LepR in the adenohypophysial cells indicate that Lep may regulate gonadotroph function through autocrine/paracrine pathway in the bovine adenohypophysis.

The roles of microtubules and microfilaments on distribution of mitochondria were evaluated by using fluorescent staining in 2-cell embryos of golden hamsters with or without cytoskeletal assembly inhibitors. In 2-cell embryos without treatment (control), most mitochondria were accumulated at the perinuclear region, while some mitochondria were noted at the cell cortex. Microtubules were found around the nuclei, correlating with distribution of the mitochondria. In contrast, microfilaments were stained intensely beneath the cell membrane and especially at the cell-to-cell contact region. In most (82%) of embryos treated with nocodazole (an inhibitor of microtubule polymerization), mitochondria had extended into the subcortical (intermediate) region with varying degree, where they were aggregated in patches. After a treatment of cytochalasin D (an inhibitor of actin polymerization), distributional density of mitochondria decreased at the cell cortex, suggesting that mitochondria moved back around the nucleus. After a treatment of both inhibitors, the distribution pattern of mitochondria was almost similar to that observed after cytochalasin D treatment. Our results suggest that the translocation of mitochondria to the perinuclear region is mediated by microtubules, while the movement of mitochondria to the cell cortex is regulated by microfilaments. Microtubules and microfilaments may function as bidirectional anchors of mitochondria to the perinuclear region and to the peripheral region, respectively.

The purpose of this study was to evaluate effect of cooling on the cytoskeletal organization affecting the distribution and structure of the meiotic spindle in the porcine metaphase II (MII) oocyte. Fluorescence staining was done for visualization of microtubules, microfilaments and chromosomes. In vitro-matured oocytes were either maintained at 37°C (for controls) or cooled abruptly to 5 or 18 ° C. Microtubules were preferentially labeled at the meiotic spindle and microfilaments were mainly detected at the cortex of the MII oocyte. The majority of controls (87%, n=97) had a barrel-shaped spindle, whereas the remaining 13% showed a broad, box-shaped spindle. After exposure to 18°C for 30 min, 94% of oocytes (n=48) possessed either a box-shaped spindle (63%) with a shortened interpolar distance (7.4 ± 0.4 µm vs. 9.8 ± 0.2 µm for controls P

The objective of this study was to monitor changes in the pyridinoline concentration of collagen of the fast-twitch gastrocnemius and slow-twitch soleus muscles of Japanese Saanen male goat kids during the period from 2 weeks before birth to 24 weeks of age. The moisture concentrations of both muscles decreased and the crude protein concentration increased steadily throughout the experimental period. The percentage of total collagen in the muscular protein showed a marked decrease (80.6–41.8% in the gastrocnemius and 77.9–40.5% in the soleus muscle) during the 2 week prenatal period. Similarly, there was a decrease in soluble collagen concentration (27.8–11.6% in the gastrocnemius and 32.6–18.1% in the soleus muscle) during the prenatal period, but the decrease in total and soluble collagen concentration was slight thereafter. There was no clear tendency for change in collagen heat solubility of both muscles, and no strong relationship was identified between collagen heat solubility and pyridinoline concentration. Pyridinoline concentration in total collagen increased during the prepubertal period in both muscles, but the soleus muscle increased faster and had a higher concentration of pyridinoline (0.22 mol/mol collagen at 20 weeks of age) than the gastrocnemius muscle (0.11 mol/mol collagen at 20 weeks of age). It was found that the increase in pyridinoline concentration began during the prenatal period, and the development of a cross-linking with age was faster in the soleus than in the gastrocnemius muscle, without collagen concentration between the muscles. It is suggests that meat toughness would be improved when the proportion of fast-twitch muscular fibrils could be increased.

Limiting maternal nutrient intake during ovine adolescent pregnancy progressively depleted maternal body reserves, impaired fetal nutrient supply, and slowed fetal soft tissue growth. The present study examined placental growth, angiogenic gene expression, and vascular development in this undernourished adolescent model at Days 90 and 130 of gestation. Singleton pregnancies were established, and ewes were offered an optimal control (C; n = 14) or low (L [0.7 3 C]; n = 21) dietary intake. Seven ewes receiving L intakes were switched to C intakes on Day 90 of gestation (L-C). Fetal body weight (P < 0.01) and glucose concentrations (P < 0.03) were reduced in L versus C pregnancies by Day 130, whereas L-C group values were intermediate. Placental cellular proliferation, gross morphology, and mass were independent of maternal nutrition at both Day 90 and 130. In contrast, capillary area density in the maternal caruncular portion of the placentome was reduced by 20% (P < 0.001) at both stages of gestation in L compared with C groups. Caruncular capillary area density was equivalent in the L and L-C groups at Day 130. Placental mRNA expression of five key angiogenic ligands or receptors increased (P < 0.001) between Days 90 and 130 of gestation. VEGFA mRNA expression was higher (P < 0.04) in L compared with C and L-C pregnancies at Day 130, but otherwise gene expression of the remaining angiogenic factors and receptors analyzed was unaffected by maternal intake. Undernourishing the pregnant adolescent dam restricts fetal growth independently of changes in placental mass. Alterations in maternal placental vascular development may, however, play a role in mediating the previously reported reduction in maternal and hence fetal nutrient supply. adolescent pregnancy, angiogenesis, female reproductive tract, fetus, placenta, placental vascular development, pregnancy, progesterone, undernutrition

The influence of relative maternal undernutrition on growth, endocrinology, and metabolic status in the adolescent ewe and her fetus were investigated at Days 90 and 130 of gestation. Singleton pregnancies to a single sire were established, and thereafter ewes were offered an optimal control (C; n = 14) or low (L [0.7 x C]; n = 21) dietary intake. Seven ewes receiving the L intake were switched to the C intake on Day 90 of gestation (L-C). At Day 90, live weight and adiposity score were reduced (P < 0.001) in L versus C dams. Plasma insulin and IGF1 concentrations were decreased (P < 0.02), whereas glucose concentrations were preserved in L relative to C intake dams. Fetal and placental mass was independent of maternal nutrition at this stage. By Day 130 of gestation, when compared to C and L-C dams, maternal adiposity was further depleted in L intake dams; concentrations of insulin, IGF1, and glucose were reduced; and nonesterified fatty acids increased. At Day 130, placental mass remained independent of maternal nutrition, but body weight was reduced (P < 0.01) in L compared with C fetuses (3555 g vs. 4273 g). Body weight was intermediate (3836 g) in L-C fetuses. Plasma glucose (P < 0.03), insulin (P < 0.07), and total liver glycogen content (P < 0.04) were attenuated in L fetuses. Fetal carcass analyses revealed absolute reductions (P < 0.05) in dry matter, crude protein, and fat, and a relative (g/kg) increase in carcass ash (P < 0.01) in L compared with C fetuses. Thus, limiting maternal intake during adolescent pregnancy gradually depleted maternal body reserves, impaired fetal nutrient supply, and slowed fetal soft tissue growth.

We measured the growth performance and meat quality of 10 crossbred (Yorkshire × Duroc × Landrace) neutered male pigs to evaluate the effects of apple pomace-mixed silage (APMS). The pigs were divided into two groups and were respectively fed the control feed and the AMPS ad libitum during the experiment. No difference was found in the finished body weight, average daily gain, carcass weight, back fat thickness or dressing ratio between the control and the AMPS treatments, but average dairy feed intake (dry matter) was significantly lower and feed efficiency was significantly higher using the APMS treatment (P 0.05). With regard to meat quality, the APMS increased the moisture content but decreased the water holding capacity (P 0.05) compared with the control treatment. Furthermore, the APMS affected the fatty acid composition of the back fat by increasing linoleic acid (C18:2n6), linolenic acid (C18:3) and arachidic acid (C20:0) levels, while decreasing palmitic acid (C16:0), palmitoleic acid (C16:1) and heptadecenoic acid (C17:1) levels, compared with the control treatment. These results indicate that feeding fermented apple pomace to finishing pigs increases the feed efficiency and affects the meat quality and fatty acid composition of back fat.

Collagen content and architecture of the longissimus thoracis (LT) muscle were compared between the concentrate- and forage-fed male goats. During the experimental period of 9 to 26 wks old, the former gained 15.7kg in body weight by feeding on commercial concentrate food and Italian ryegrass hay, while the latter gaincd only 2.4 kg by feeding on hay only. Dry matter, crude protein and crude fat were contained at significantly higher percentages in the LT muscle of the concentrate-fed goats than the forage-fed goats, and conversely, collagen content was significantly higher in the muscle of the latter. However, the collagen pyridinoline content did not differ between them. The myofibers in the concentrate-fed goats (54.0~am) were twlce the size of those in the forage-fed goats (25.5llm). The higher content of crude fat (2.890/0) in the LT muscle of the concentrate-fed g6ats was not enough for adipose tissues to develop wlthin the perimysium as marbling. Immunohistochemical methods revealed types I and HI collagens over both perimysium and endomysium. Thin collagen bundles were loosely distributed in the perimysium of concentrate-fed goats muscles, and conversely, the perimysial architecture in the forage-fed goats muscles appeared as dense stacks of thick collagen fibers. From these results, it was indicated that the development of loose tissues of slender collagen fibers had an intimate relationship with the rapid growih rate of muscle rather than the development of adipose tissue.

We investigated testicular and circulating levels of dimeric inhibins in Holstein bulls from the infantile to postpubertal periods (5 to 50 weeks of age) and examined the relationship between the profiles of circulating dimeric inhibins and FSH. Concentrations of total inhibin and inhibin B in the testis were highest at 4 to 5 weeks of age but decreased gradually as the bulls aged. Testicular inhibin A levels showed a gradual decline to a nadir at 15 to 26 weeks of age, but by 39 weeks, they were high again. The contents of total inhibin, inhibin A, and inhibin B per testis generally increased with age. Fractionation of testicular homogenates obtained from 15-week-old bulls by a combination of immunoaffinity chromatography and SDS-PAGE confirmed the presence of two major molecular weight forms (32 and 45 kDa) of dimeric inhibins in the testes. Circulating levels of total inhibin and inhibin A showed a significant increase in bulls at around 10 to 14 weeks of age compared to the levels between 5 and 7 weeks of age but decreased thereafter. However, immunoreactivity for inhibin B was not detected in the peripheral circulation, probably because of low sensitivity of the inhibin B assays. The concentrations of plasma FSH were high at 5 weeks of age but declined to lower levels between 11 and 40 weeks, and then increased from 41 weeks onward. There was no significant correlation between the plasma levels of FSH and inhibin A or total inhibin. The results clearly indicate that the bull testis produces inhibin A and B and secretes at least inhibin A into the circulation during postnatal development. However, the profile of circulating FSH in bulls shows no reciprocal relationship with the inhibin A or total inhibin profile during the postnatal period.

Effects of preslaughter nutritional condition on intramuscular collagen characteristics were studied, in order to clarify the potential of intensive feeding to aged animals in improving meat tenderness. Ten castrated male goats were assigned into one of two groups: one group was allowed ad libitum access to a concentrate diet (total digestible nutrients 70%, crude protein 15%) and Italian ryegrass hay (ADLIB-group), and the other group was fed a restricted amount of their diet (concentrate diet 0.5% of bodyweight/day; hay 1.5% of bodyweight/day) to maintain their bodyweight (MAIN-group). After 3 months of the experimental period, goats were slaughtered and meat samples were obtained immediately. Goats in ADLIB-group had lower total and insoluble collagen concentrations, higher fat concentrations and collagen solubility than those in MAIN-group, but soluble collagen concentrations of muscles were similar for both groups. Goats in ADLIB-group had lower pyridinoline concentrations than those in MAIN-group, in all muscles, but the differences of pyridinoline concentration between the groups were not statistically significant. Warner-Bratzler shear force values of Longissimus and Biceps femoris muscles were lower in ADLIB-group than in MAIN-group. The increase of meat tenderness by preslaughter intensive feeding seemed to be associated with the increase in intramuscular fat deposition and high collagen solubility, and pyridinoline cross-link appeared to be one of the factors related to collagen solubility.

Pituitary transcription factor-1 (Pit-1) is a tissue-specific transcription factor responsible for somatotroph (GH cell) development and growth hormone (GH) gene activation. To investigate the expression of Pit-1 protein in bovine adenohypophysis, antigen retrieval conditions for detection of Pit-1 were optimized in paraffin sections of bovine pituitary. We then localized Pit-1 and GH employing a double-immunohistochemical staining method and compared the distribution and proportion of adenohypophyseal cells containing Pit-1 and GH between Japanese Black (Blk) and Holstein (Hol) steers. Autoclave heating pretreatment of sections in Dako Target Retrieval Solution for 20‐30 min enabled reliable detection of the distinct nuclear immunostaining of Pit-1. However, heat treatment in phosphate buffered saline or citrate buffer gave only a weak signal with increased background staining and false positive immunoreactions in the cytoplasm. Pituitary transcription factor-1 was abundantly expressed in the bovine anterior pituitary and the number of Pit-1 immunoreactive cells did not differ between the breeds. The proportion of GH cells relative to Pit-1 immunoreactive cells was smaller in Blk than Hol steers, suggesting an intrinsic breed difference in cellular composition of bovine adenohypophysis. The immunohistochemical technique described will facilitate further investigation of the role played by Pit-1 in the regulation of GH cell development and function in cattle.

褐毛和種去勢牛12頭を，アマニ油脂肪酸カルシウムを4%添加した濃厚飼料を給与する油脂区（6頭）と無添加の濃厚飼料を給与する対照区（6頭）に分け，14ヵ月齢から10ヵ月間肥育を行い，この間のメタン発生と肥育成績を比較検討した．乾物摂取量（DMI）あたりのメタン発生量は，有意な差ではなったが，試験期間を通じて対照区よりも油脂区で小さな値を示した．また，アマニ油脂肪酸カルシウム添加によるメタン発生の減少量は不飽和脂肪酸摂取量から計算される減少量の7割程度であったことから，アマニ油脂肪酸カルシウムはメタン発生抑制に効果があったと考えられた．油脂区は対照区よりも増体成績が優れ，日増体量あたりのメタン発生量は，有意な差ではないものの13%低い値を示した．以上の結果からアマニ油脂肪酸カルシウムの添加は，DMIあたりのメタン発生を抑制し，増体成績を改善することにより，増体あたりのメタン発生量を削減する可能性が示唆された．

To examine the possible link between endocrine status and perinatal problems related to cattle cloning, plasma concentrations of cortisol, adrenocorticotropic hormone (ACTH) and components of the insulin-like growth factor (IGF) system were compared between 13 somatic cell cloned and seven control Japanese Black calves (five produced by artificial insemination [AI] and two produced from in vitro fertilized embryos [IVP]) immediately after birth. Five cloned calves required delivery by cesarean section (C-section), while all of control calves were delivered by spontaneous vaginal delivery. The C-section delivered clones were heavier at birth, followed by vaginally delivered clones and IVP controls, and AI controls were the lightest. The neonatal mortality (death within the 1st week) of C-section delivered clones was also high (4/5) compared to that of vaginally delivered clones (1/8) or controls (0/7). Plasma concentrations of cortisol and IGF-I were lower in the clones than control calves although the plasma ACTH level was not different between the groups. A striking difference was observed in plasma IGF binding protein (IGFBP) profile in which cloned calves had a greater relative abundance of IGFBP-2 compared with controls. Observed differences suggest that insufficient prepartum rise in plasma cortisol of cloned calves failed to initiate the switch to an adult mode of the IGF system during late gestation and therefore parturition was not spontaneous. Inappropriate developmental changes in endocrine system may be partly responsible for the fetal overgrowth and perinatal complications associated with the cloning technology.

1. Histological and histochemical profiles of Musculus pectoralis (PT, type IIB fibres), M. iliotibialis lateralis (ITL, types IIA + IIB fibres) and M. puboischiofemoralis pars medialis (PIF, type I fibres) were compared in carbon dioxide (37%, 70 s) and electrically (14 V, 5 s) stunned male chickens. 2. Muscle materials were taken at 0, 4 and 24 h from carcases dressed and cooled with ice-water mixture for 30 min. Glycogen and fat contents, and adenosine triphosphatase and reduced nicotinamide adenine dinucleotide dehydrogenase activities of fibres were measured. 3. In PT muscle at 0 h, gas stunned chickens showed many fibres with high glycogen content but those electrically stunned contained few such fibres. Fibres from gas stunned birds had lost almost all their glycogen after 24 h of cold storage. 4. In the ITL muscle of gas stunned chickens at 0 h residual glycogen was observed in type IIB fibres. In contrast, in the electrically stunned birds it was in type IIA, showing the different effects of the stunning methods. During cold storage, glycogen disappeared earlier in type IIB than IIA fibres. 5. In PIF muscle with fibres of low glycogen content, the gas stunned chickens maintained a good fibre structure for 4 h or more, but the electrically stunned had already lost intact fibre structure at 4 h. 6. These results indicated that the carbon dioxide stunning was a better method for chicken welfare and meat quality than electrical stunning.

Japanese Black cattle (Wagyu), deposit much higher amounts of intramuscular fat, known as marbling, than other breeds of cattle. To determine whether this unique fat deposition is attributable to the somatotropic axis, we compared pulsatile growth hormone (GH) secretion, plasma levels of insulin-like growth factor-1 (IGF-1) and cellular density of somatotrophs (GH-expressing cells) in the anterior pituitary glands of Japanese Black and Holstein steers. Blood samples were withdrawn every 15 min for 6 h from 14 Japanese Black and 12 Holstein steers at about 17 months of age, and GH and IGF-1 concentrations were determined. The distribution and proportion of GH-expressing cells were analysed by immunohistochemistry combined with point-count morphometry in pituitaries from six steers from each breed aged about 18 to 21 months. Overall mean and baseline plasma GH concentrations were lower (P < 0·001) in Japanese Black than Holstein steers. In addition, Japanese Black had smaller (P < 0·05) amplitudes of GH secretory pulses than Holstein steers, whereas the GH pulse frequency did not differ between the breeds. Japanese Black steers also had lower (P < 0·001) plasma levels of IGF-1 than Holstein steers. The marbling score of Japanese Black steers was higher (P < 0·001) than that of Holsteins at the same carcass weight. The proportion of GH-expressing cells was smaller (P < 0·05) in Japanese Black than Holstein steers at the hind dorsal and hind ventral regions of the adenohypophysis. Thus, in Japanese Black and Holstein steers, the breed difference in the relative density of GH-expressing cells corresponded to that in profiles of pulsatile GH secretion. These results suggest that the features of the somatotropic axis intrinsically differ between Japanese Black and Holstein cattle and that these features may be partly responsible for the genetic ability of the former to deposit greater amounts of marbling fat and for the smaller frame of Wagyu cattle.

The relationship between desaturation of fatty acid and conjugated linoleic acid (CLA) accumulation in ruminants body fat was examined. Ten castrated goats were assigned to one of two treatments, maintenance group and full-feed group. After a 90 day feeding trial, subcutaneous fat, perinephric fat, Musculus longissimus thoracis, digesta in rumen and abomasum, Medulla ossium flava in left Ossa metatarsalia, Ossa cruris and Femur were obtained. The unsaturated fatty acid and CLA concentration in subcutaneous fat were significantly higher compared to the perinephric fat, and the values of the full-feed group were higher than those of the maintenance group. The CLA and unsaturated fatty acid concentrations in Medulla ossium flava were the highest in Ossa metatarsalia located on body surface followed by Ossa cruris and Femur. CLA concentration in digesta was low, thus the absorption of CLA from digestive tract was not the only important route for CLA supply to ruminant body fat. A high positive correlation between C17: 1/C17: 0 and CLA/t11-C18:1 was recognized in adipose tissues and Medulla ossium flava. The results agreed with the hypothesis that a greater part of CLA in ruminant adipose tissues was biosynthesized. First, the poly-unsaturated fatty acid in feedstuff is changed into t11-C18:1 by rumen biohydrogenation and then t11-C18:1 is absorbed from digestive tract and accumulated in the adipose tissues before converting into CLA by desaturation.

The distribution and proportion of somatotrophs (GH cells) and mammotrophs (PRL cells) were immunohistochemically detailed in the adenohypophysis of pure bred of Japanese Black steers. The animals were divided into four groups of six each of 8, 14, 21 and 28 months-old. The morphometrical analysis was evaluated in four regions of the midsagittal plane - zona tuberalis (ZT), and hind dorsal (HD), fore ventral (FV) and hind ventral (HV) regions. GH cells mainly existed solitarily and densely populated in the HD, and PRL cells were numerously distributed in the FV and HV as various sizes of cell clusters, whereas, in the ZT, the proportion of both GH cells and PRL cells was very low. The proportion of GH cells gradually diminished until 21 months-old and that of PRL cells notably decreased until 28 months-old. However, the distribution pattern of GH cells and PRL cells did not change during 8 to 28 months-old. These results initially showed that GH cells and PRL cells were differentially distributed in the whole adenohypophysis of Japanese Black steers and the proportion declined age-dependently.

The existence and distribution of mammosomatotrophs (MS cells) containing growth hormone (GH) and prolactin (PRL) were detailed in swine adenohypophysis. MS cells were positively identified and were first recognized in the adenohypophysis of fetuses with 33.0 cm in body length, when GH cells existed earlier and PRL cells occurred. The proportion of MS cells transiently increased in the glands of three month-old females and were densely populated in the fore ventral (FV) and the hind ventral (HV) region except for the zona tuberalis (ZT). The distribution was similar to GH cells and PRL cells. MS cells were divided into three subtypes by the intracellular intensity of both GH and PRL stainings. Type I MS cells which were strongly stained with anti-GH antiserum, constituted 100% of MS cells in fetuses and the proportion decreased with age in females. Type II MS cells which were evenly stained with both anti-GH and PRL antisera were detected in all animals except for fetuses, and were proportioned moderately. Type III MS cells which contained more PRL than GH existed predominately in three and six month-old females, castrated males and aged males. The findings indicated that an amount of GH and PRL within MS cells was changeable in response to physiological conditions. The results suggest that GH cells may transform into PRL cells, with the MS cell acting as an intermediate cell in swine adenohypophysis, and that the GH and PRL in MS cells are synthesized synchronously or asynchronously according to the secretory activities.

In the present study, we investigated conjugated linoleic acid(CLA) concentrations in the adipose tissues from carcasses of Japanese Black fattening steers and was correlated with the roughage intake. Subcutaneous, intermuscular and peninephric fats were collected from 8 Japanese Black fattening steers with mean body weight of 670±79kg. The animals were fed with concentrate and chopped rice straw ad libitum for 11 months, from 15 to 26 months of age. Conjugated linoleic acid concentrations in subcutaneous, intermuscular and perinephric fats were 0.44, 0.34 and 0.25%, respectively. The difference in mean CLA concentrations among the adipose tissues was significant (P

A specific monoclonal antibody, M3A, was produced to rapidly purifyLactococcus lactisphosphoglucose isomerase (PGI) for amino acid sequence analysis. M3A recognized theLac. lactisPGI specifically and sensitively with both enzyme-linked immunosorbent assay and Western blot analysis. The enzyme was rapidly purified to a specific activity of 21.8 U/mg with a yield of 20% by a three-step procedure, including M3A-bound Sepharose chromatography. The specific activity of PGI was increased about 64.1-fold from the cell lysate. The molecular mass ofLac. lactisPGI was estimated to be about 50 kDa by SDS–PAGE. The N-terminal amino acid sequence ofLac. lactisPGI exhibited no significant similarity to other PGIs, except for a 52.6% identity toBacillus stearothermophilusPGI A and PGI B. These results suggest that there might be some molecular types of PGI.

The objective was to evaluate effects of feeding level of milk replacer on body growth, plasma metabolite and insulin concentrations, and allometric growth of visceral organs in suckling calves. Holstein bull calves (n = 8; 3-4 days of age) were fed either a low amount (average 0.63 kgDM/day, LM) or high amount (average 1.15 kgDM/day, HM) of high protein milk replacer until they were slaughtered at 6 weeks of age. Body weight (BW) at 4, 5, and 6 weeks of age, feed intake, average daily gain, and feed efficiency were higher in the HM than LM calves. The HM group had higher plasma glucose at 3 and 4 weeks of age and insulin levels after the age of 4 weeks compared with LM calves whereas no effect was detected on plasma nonesterified fatty acid or urea nitrogen concentrations. The HM calves had greater empty body weight (EBW), viscera-free BW and most of the organs dissected than LM calves. Relative weights (% of EBW) of liver, spleen, kidneys, and internal fat were higher, whereas head and large intestine was lower in HM than LM calves. The results suggest that increased milk feeding levels would accelerate the growth of the body and specific organs.

To improve culture system for in vitro maturation (IVM) of porcine oocytes, ghrelin, leptin or growth hormone (GH), at concentration of 0, 0.5, 5, 50 and 500 ng/ml were added to the porcine follicular fluid (pFF)-supplemented medium NCSU23, and their effects on the maturation and cytoskeletal distribution of the oocytes with or without cumulus cells were compared. In the cumulus-denuded oocytes, no significant changes were noted in the maturation rate by different hormone treatments due to a marked decline in the controls. Maturation of the cumulus intact oocytes was moderately interfered by ghrelin (0.5-50 ng/ml, p < 0.01), but not significantly affected by leptin and GH. Distribution density of the cytoplasmic microtubules was decreased significantly by addition of ghrelin (by approximately 30% in 50-500 ng/ml, p < 0.01), whereas no remarkable effect was noted by leptin supplementation. High concentration (500 ng/ml) of ghrelin or leptin decreased significantly the cytoplasmic microfilaments in density (by 43% and 38%, p < 0.01, respectively). GH did not affect cytoskeletal distribution. The results suggest, in the culture system using pFF-supplemented medium that (i) ghrelin may have some inhibitory effect on the organization of microtubules and microfilaments, probably being a factor in lowered maturation rate and (ii) the addition of higher concentration of leptin may decrease microfilaments in density with no effect on meiotic maturation of the porcine oocytes.

The influence of relative maternal undernutrition on growth, endocrinology, and metabolic status in the adolescent ewe and her fetus were investigated at Days 90 and 130 of gestation. Singleton pregnancies to a single sire were established, and thereafter ewes were offered an optimal control (C; n = 14) or low (L [0.7 x C]; n = 21) dietary intake. Seven ewes receiving the L intake were switched to the C intake on Day 90 of gestation (L-C). At Day 90, live weight and adiposity score were reduced (P0.001) in L versus C dams. Plasma insulin and IGF1 concentrations were decreased (P0.02), whereas glucose concentrations were preserved in L relative to C intake dams. Fetal and placental mass was independent of maternal nutrition at this stage. By Day 130 of gestation, when compared to C and L-C dams, maternal adiposity was further depleted in L intake dams; concentrations of insulin, IGF1, and glucose were reduced; and nonesterified fatty acids increased. At Day 130, placental mass remained independent of maternal nutrition, but body weight was reduced (P0.01) in L compared with C fetuses (3555 g vs. 4273 g). Body weight was intermediate (3836 g) in L-C fetuses. Plasma glucose (P0.03), insulin (P0.07), and total liver glycogen content (P0.04) were attenuated in L fetuses. Fetal carcass analyses revealed absolute reductions (P0.05) in dry matter, crude protein, and fat, and a relative (g/kg) increase in carcass ash (P0.01) in L compared with C fetuses. Thus, limiting maternal intake during adolescent pregnancy gradually depleted maternal body reserves, impaired fetal nutrient supply, and slowed fetal soft tissue growth.

Limiting maternal nutrient intake during ovine adolescent pregnancy progressively depleted maternal body reserves, impaired fetal nutrient supply, and slowed fetal soft tissue growth. The present study examined placental growth, angiogenic gene expression, and vascular development in this undernourished adolescent model at Days 90 and 130 of gestation. Singleton pregnancies were established, and ewes were offered an optimal control (C; n = 14) or low (L [0.7 x C]; n = 21) dietary intake. Seven ewes receiving L intakes were switched to C intakes on Day 90 of gestation (L-C). Fetal body weight (P0.01) and glucose concentrations (P0.03) were reduced in L versus C pregnancies by Day 130, whereas L-C group values were intermediate. Placental cellular proliferation, gross morphology, and mass were independent of maternal nutrition at both Day 90 and 130. In contrast, capillary area density in the maternal caruncular portion of the placentome was reduced by 20% (P0.001) at both stages of gestation in L compared with C groups. Caruncular capillary area density was equivalent in the L and L-C groups at Day 130. Placental mRNA expression of five key angiogenic ligands or receptors increased (P0.001) between Days 90 and 130 of gestation. VEGFA mRNA expression was higher (P0.04) in L compared with C and L-C pregnancies at Day 130, but otherwise gene expression of the remaining angiogenic factors and receptors analyzed was unaffected by maternal intake. Undernourishing the pregnant adolescent dam restricts fetal growth independently of changes in placental mass. Alterations in maternal placental vascular development may, however, play a role in mediating the previously reported reduction in maternal and hence fetal nutrient supply.

In the overnourished adolescent sheep, maternal tissue synthesis is promoted at the expense of placental growth and leads to a major decrease in lamb birth weight at term. Maternal growth hormone (GH) concentrations are attenuated in these pregnancies, and it was recently demonstrated that exogenous GH administration throughout the period of placental proliferation stimulates uteroplacental and fetal development by Day 81 of gestation. The present study aimed to determine whether these effects persist to term and to establish whether GH affects fetal growth and body composition by increasing placental size or by altering maternal metabolism. Adolescent recipient ewes were implanted with singleton embryos on Day 4 postestrus. Three groups of ewes offered a high dietary intake were injected twice daily with recombinant bovine GH from Days 35 to 65 of gestation (high intake plus early GH) or from Days 95 to 125 of gestation (high intake plus late GH) or remained untreated (high intake only). A fourth moderate-intake group acted as optimally nourished controls. Pregnancies were terminated at Day 130 of gestation (6 per group) or were allowed to progress to term (8-10 per group). GH administration elevated maternal plasma concentrations of GH, insulin, glucose, and nonesterified fatty acids during the defined treatment windows, while urea concentrations were decreased. At Day 130, GH treatment had reduced the maternal adiposity score, percentage of fat in the carcass, and internal fat depots and leptin concentrations, predominantly in the high-intake plus late GH group. Placental weight was lower in high-intake vs. control dams but independent of GH treatment. In contrast, fetal weight was elevated by late GH treatment, and these fetuses had higher relative carcass fat content, perirenal fat mass, and liver glycogen concentrations than all other groups. Expression of leptin mRNA in fetal perirenal fat and fetal plasma leptin concentrations were not significantly altered by maternal nutritional intake or GH. In pregnancies proceeding to term, the duration of gestation, fetal placental mass, and lamb birth weight were reduced in high-intake compared with control dams but were not significantly affected by GH treatment. In conclusion, exogenous GH has profound effects on maternal endocrinology, metabolism, and body composition when administered during early and late pregnancy. Treatment during late pregnancy has a modest effect on fetal growth independent of placental size and a profound effect on fetal adiposity, which may have implications beyond the fetal period.

A pro-inflammatory cytokine, interleukin 18 (IL-18), induces intracellular expression of IL-1 and the release of IL-6. IL-1 and IL-6 has been detected in anterior pituitary cells, suggesting that IL-18 is produced in anterior pituitary cells and may serve to aid immuno-endocrine regulation. In the present study, we addressed this hypothesis by investigating the intracellular localization of IL-18 and its receptor in bovine anterior pituitary gland. IL-18 mRNA and its protein were detected in the anterior pituitary gland by RT-PCR and Western blotting. In situ hybridization showed that IL-18 mRNA was expressed in the anterior pituitary cells. Immunohistochemistry of IL-18 and specific hormones revealed the presence of IL-18 in somatotrophs. Furthermore, the expression of GH mRNA in IL-18 immunoreactive cells was confirmed by immuno-laser microdissection. These results first demonstrated that somatotrophs produced IL-18. Subsequently, the distribution of the IL-18 receptor alpha (IL-18Ralpha) was investigated in order to understand IL-18 signaling among the anterior pituitary cells. Bovine IL-18Ralpha cDNA was partially sequenced and detected in the anterior pituitary gland by RT-PCR. Immunohistochemistry of IL-18Ralpha, IL-18 and GH showed that IL-18Ralpha was co-localized in IL-18 immunoreactive cells or somatotrophs. These data suggest that IL-18 acts on somatotrophs as an immuno-endocrine mediator through the autocrine pathway.

The existence and distribution of mammosomatotrophs (MS cells) containing growth hormone (GH) and prolactin (PRL) in bovine adenohypophysis were detailed by a combined method of mirror sections and immunohistochemical staining. MS cells always occurred in bovine adenohypophysis but their number was low. In the midsagittal plane, the cells were observed in the hind dorsal, hind ventral and fore ventral region abundant in GH and PRL cells. Whereas, in the zona tuberalis where GH and PRL cells were less frequent, MS cells were not detected. MS cells were invariably solitarily distributed within mammotroph (PRL cell) clusters but not within somatotroph (GH cell) clusters. The proportion of MS cells declined as the ages proceeded and the appearance was spatially related to the arrangement of PRL cells. These findings indicated that, in bovine adenohypophysis, MS cells were differentially distributed and occurred in PRL cell clusters. The results strongly suggest that MS cells originate in GH cells pre-existed within PRL cell clusters with special reference to the functional activation of PRL cells.

Overnourishing the adolescent sheep promotes rapid maternal growth at the expense of the gravid uterus. The growth of the placenta is impaired and results in the premature delivery of low-birthweight lambs. The present study details fetal adipose tissue development in these growth-restricted pregnancies. Singleton pregnancies were established by embryo transfer and, thereafter, adolescent ewes were offered a high (H; n = 12) or moderate (M; n = 14) level of a complete diet until necropsy on Day 131 of gestation. Fetal weight was lower (P < 0.001) in H compared with M groups. High maternal intake preserved brain and perirenal fat weight (P < 0.003), whereas relative weights of the heart, lungs, spleen and liver were unaltered. High nutrient intake resulted in significantly elevated maternal plasma concentrations of insulin, leptin, prolactin and glucose, no significant changes in fetal insulin, leptin or non-esterified fatty acids and attenuated fetal prolactin concentrations. Irrespective of nutritional intake, maternal plasma leptin, prolactin and glucose concentrations were negatively correlated with fetal weight and were positively correlated with fetal perirenal fat proportion (all P < 0.01). The mRNA expression for leptin, prolactin receptor and uncoupling protein (UCP) 1 in fetal perirenal fat was equivalent between groups, but, irrespective of maternal nutrition, UCP1 mRNA levels were negatively correlated with fetal weight (P < 0.01). Thus, overnourishing pregnant adolescent sheep preserves fat deposition in their growth-restricted fetuses, which may have implications for neonatal thermogenesis and for programming of postnatal adiposity.

Growth hormone (GH) is a vital component for the regulation of animal growth. The circulating levels of GH decrease with age in bull calves (McAndrews et al., 1993) and in steers (Verde and Trenkle, 1987), and are higher in large Simmental steers than in small Angus steers (Grigsby and Trenkle, 1986). Japanese Black cattle, known as Wagyu, is early-matured and small frame size, and has an ability to deposit the high amount of intramuscular fat. The present study was conducted to clarify the age-related changes of GH secretion and somatotroph function in Japanese Black steers, since the information about endocrine profiles of Japanese Black cattle has been scantly.

The LPE Growth of In1-x Ga x P1-z As z (z ≦0.01) on (1 0 0) GaAs substrates is described with stress on the relation between the compositions of source melts and the quality of grown crystals. The melt in equilibrium with In1-x Ga x P0.99As0.01 lattice-matched to GaAs at 785°C is found to consist of 0.87 at.% of Ga, 2.66 at.% of P, 0.05 at.% of As and the rest In. Lattice constants strained in the direction of growth of the layers grown from melts supersaturated by 4°C were pinned near to a value 0.3% larger than that of GaAs substrates. However, the pinning was not observed for wafers grown from melts supersaturated by more than 8.6°C. The photoluminescence of the wafers is also reported.

An LPE growth method of In1-x Ga x P y As1-y layers on (1, 0, 0) GaAs is described over a wide range of band-gap energy. Melt compositions for lattice-matched growth of these layers are given. Photoluminescence spectra of the layers are reported, and their spectral shape and luminescent-peak energies are discussed in comparison with In1-x Ga x P y As1-y grown on InP.