Expression levels of FSHR, IGF1R, CYP11al and HSD3ß in cumulus cells can predict in vitro developmental competence of bovine oocytes.

The efficiency of in vitro embryo production technologies would be improved by the development of suitable non-invasive biomarkers that allow the selection of good quality cumulus-oocyte complexes (COCs). The present study used whole, single oocyte culture to investigate whether the expression levels of follicle-stimulating hormone receptor (FSHR), insulin-like factor 1 receptor (IGF1R) and three steroidogenesis-related enzymes (CYP11al, CYP19al and HSD3ß) in cumulus cells reflected the developmental competence of COCs. Cumulus cells were collected from single COCs before maturation culture and relative mRNA levels were assessed using real-time PCR. The analysis indicated that mRNAs for FSHR, IGF1R, CYP11al and HSD3ß were present at higher levels in cumulus cells from COCs that failed to form blastocysts compared with cumulus cells from COCs that formed blastocysts. Moreover, FSHR and IGF1R mRNA levels were positively correlated with those of genes for steroidogenesis-related enzymes. In conclusion, poor developmental competence of COCs was related to higher expression of FSHR, IGF1R, CYP11al and HSD3ß in cumulus cells, which may indicate the advanced differentiation of cumulus cells into granulosa cells.

Effects of a tunnel ventilation system within the tie-stall barn environment upon the productivity of dairy cattle during the winter season.

OBJECTIVE: The objective of this study was to examine the effect of using a tunnel ventilation system within the dairy barn environment upon the productivity of dairy cows during the winter season. METHODS: The study was performed at the University Farm, Faculty of Agriculture, Utsunomiya University. Twenty-one Holstein dairy cows (5 heifers and 16 multiparous) were enclosed in a stall barn. Unventilated (UV) and tunnel-ventilated (TV) was operated by turns every other week, and a number of key parameters were measured in the barn, including tunnel ventilation output, temperature, relative humidity, gas concentrations (oxygen [O2], carbon dioxide [CO2], and ammonia [NH3]). Also, skin and rectal temperature, respiratory rate, blood gas concentrations, and bacterial count were measured from nipple attachments on ten cows. The amount of fodder left uneaten, and general components and somatic cell count of the milk were measured. RESULTS: As for our dairy barn environment, air temperature dropped significantly with the passage of time with TV. Humidity was significantly higher with TV at 0600 h compared to UV, while CO2 and NH3 concentrations with UV were significantly higher than with TV at 0000 h and 0600 h. Skin temperature was significantly lower with TV compared to UV at 0000 h and 0600 h. Respiratory rate was also significantly lower at 0600 h with TV than with UV. Bacterial count for the nipple attachments was significantly lower with TV than with UV at 0600 h. The amount of leftover fodder was significantly less with TV in comparison with UV. CONCLUSION: Our results suggest that a TV system in the winter barn results in environmental improvements, such as reductions in unfavorable gas concentrations and bacterial growth. Consequently, it is expected that barns utilizing a TV system will be beneficial for both animal health and production.

Effects of insulin-like growth factor-1 on the in vitro maturation of canine oocytes.

The maturation rate of canine oocytes during in vitro maturation (IVM) needs to be improved. The present study was designed to evaluate the effects of insulin-like growth factor-1 (IGF-1) on the IVM of canine oocytes. Ovaries were obtained by ovariohysterectomy and were sliced to release cumulus-oocyte complexes (COCs). In Experiment 1, the effects of different concentrations of IGF-1 on the nuclear maturation of oocytes was investigated. The COCs were cultured in a modified medium (mTCM199) with IGF-1 (0, 0.5, 5, 10, and 50 µg/ml). At the end of the 48 h culture, oocytes were fixed and stained to evaluate their nuclear stage. Supplementation with 50 µg/ml IGF-1 induced a significantly higher metaphase II (MII) rate (P < 0.05) compared to the 0 and 0.5 µg/ml IGF-1 groups. In Experiment 2, the expression levels of insulin receptor (INSR), IGF-1 receptor (IGF-1R), and IGF-2 receptor (IGF-2R) genes, localized to canine oocytes and cumulus cells, were investigated before and after IVM. The expression level of IGF-1R in cumulus cells after IVM was higher than that before IVM (P < 0.05). In Experiment 3, it was investigated whether an inhibitor of PTEN (phosphatase and tensin homolog), bpV, affects the nuclear maturation of oocytes. Regardless of bpV supplementation at a concentration of 0.2 to 200 µmol/l, there was no significant difference in the proportion of oocytes that reached the MII stage. These results indicated that IGF-1 has a favorable effect on the IVM of canine oocytes, possibly through the stimulation of the Ras/MAPK pathway via IGF-1R expressed in cumulus cells.

Influence of maternal remifentanil concentration on fetal-to-maternal ratio in pregnant ewes.

PURPOSE: Maternal remifentanil infusion is used for minimally invasive fetal surgery or ex-utero intrapartum treatment. The fetal-to-maternal (F/M) ratio of remifentanil concentration at various dosing regimens is useful to manage remifentanil effects. The aim of this study was to investigate the F/M ratio of remifentanil at various concentrations. METHODS: Five pregnant ewes received continuous remifentanil infusion under propofol anesthesia. The remifentanil infusion rate was increased by 0.4 µg/kg/min every 15 min. The response to tail clamping in fetuses was assessed immediately before the change of infusion rate. Arterial remifentanil concentrations in the mother and fetus were determined at each tail clamp. After observing a loss of response to tail clamping, remifentanil infusion was terminated and the concentrations were assessed. RESULTS: The median remifentanil maximum infusion rate and maternal concentration were 3.0 µg/kg/min (range 2.4-3.6) and 21.6 (range 18.0-29.9) ng/mL, respectively. During continuous infusion, the F/M ratio was 0.15 (0.07-0.17), and the slope of the linear regression for the F/M ratio versus infusion rate in each individual was -0.001 ± 0.012/µg kg min (P = 0.876 vs hypothetical value of 0). The F/M ratio at the first sampling point in the elimination phase [0.33 (0.07-0.65)] was higher (P = 0.033) than at the last sampling point during continuous infusion [0.15 (0.06-0.17)]. CONCLUSION: The F/M ratio was constant at a steady state regardless of the remifentanil concentration up to 29.9 ng/mL, and increased in the elimination phase in pregnant ewes.

Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination.

The present study was conducted to evaluate whether supplementation of semen extender with glutathione (GSH) can maintain the quality of frozen-thawed canine spermatozoa. Eighteen ejaculates were obtained from 5 dogs and placed in extender (20% egg yolk, Tris, citric acid, lactose, raffinose, antibiotics and 6.5% glycerol) containing 0 (control), 2.5, 5, 7.5 or 10 mM GSH. The samples were cooled to 4 C and then frozen in liquid nitrogen vapor. Motility parameters of the sperm were evaluated at 0, 1, 2, 3, 4, 12 and 24 h after thawing. Sperm motility was higher in the 5 mM GSH group than in the control or 2.5 and 10 mM GSH groups; this effect was observed at 1 to 24 h after thawing (P < 0.05). The 5 mM GSH group had a higher sperm viability index at 12 and 24 h after thawing compared with the other groups (P < 0.05). Acrosome integrity, evaluated at 4 h after thawing, was greater in two of the GSH-treated groups (5 and 10 mM) compared with the control. Lipid peroxidation (LP) levels immediately after thawing were lower in the 5 and 10 mM GSH groups compared with the control, while those at 12 h after thawing did not differ significantly. Frozen-thawed semen in the 5 mM GSH group was used for transcervical insemination of 4 bitches, resulting in delivery of 5 puppies from 2 bitches. These results indicate that supplementation of semen extender with 5 mM GSH was effective in improving motility, longevity and acrosomal integrity and inhibiting LP levels in post-thaw canine spermatozoa, without any adverse impacts on full-term development after transcervical insemination.

Comparison of serum very low-density lipoprotein concentrations during transition in primiparous and multiparous cows.

This study was carried out as an observational study in order to examine the difference of change in serum very low-density lipoprotein (VLDL) between primiparous and multiparous cows. Twenty-one clinically healthy cows (10 primiparous and 11 multiparous) were selected at 21 days prior to expected calving. Blood samples were collected in the morning (before feeding) on days -21, -7, 7, 21 and 56 days in milk (DIM). At 7 and 21 DIM, the serum non-esterified fatty acid concentration of multiparous cows was significantly higher than that of primiparous cows. The serum ß-hydroxybutyrate concentration was also markedly higher in multiparous cows than in primiparous cows at 21 DIM. These results suggested that the degree of negative energy balance was greater in multiparous cows than in primiparous cows during this period. In both, serum VLDL concentrations decreased at over 7 DIM, increased at 21 DIM, and then decreased at 56 DIM. On the other hand, triglyceride and total protein concentrations of VLDL in multiparous cows were significantly lower than in primiparous cows at 21 DIM. This suggests that multiparous cows have poor triglyceride secretion from the liver and that they become more susceptible to hepatic lipidosis.

Characteristics of failure of passive transfer at the herd level using the serum immunoglobulin G concentration as an indicator on dairy farms in eastern Hokkaido, Japan.

The objectives of this study were to conduct a survey of failure-of-passive-transfer (FPT) in eastern Hokkaido Japan, to evaluate the association between herd-level FPT and death and culling or treatment, and to test the effectiveness of monitoring using herd-level FPT. A total of 4,411 Holstein and Holstein-Wagyu crossbreds calves born from Holstein dams during the year beginning April 2, 2019 on 39 dairy farms were included in the study to investigate death-and-culling and the treatment rate during the first month of life, as well as rearing management up to 3 weeks of age. A subset of Holsteins (n=381) was included in the study for passive transfer and farms were diagnosed as having FPT if more than 20% of newborn calves had serum IgG levels below 10 g/L at the herd level. The prevalence of FPT (

Effects of starch content of calf starter on rumen properties and blood concentrations of metabolites and hormones in dairy calves under a high plane of milk feeding.

The objective of this study was to investigate the effects of calf starters with different starch content on rumen and blood properties in calves fed under a high plane of nutrition program. Twenty-seven Holstein calves (14 female, 13 male) were assigned to one of three starter treatment groups in a randomized block design: high-starch (41.8%) (n = 9), H; medium-starch (31.9%) (n = 9), M; low-starch (22.0%) (n = 9), L. Milk replacer on powder basis was fed up to 1.2 kg/day as fed, gradually reduced from 6 weeks of age; calves were weaned at 8 weeks of age. Each starter (up to 3.5 kg/day as fed) and chopped hay were provided ad libitum from 5 days of age. Blood was collected from 4 to 13 weeks of age. Rumen fluid was collected at 6, 8, 10, and 13 weeks of age. Serum urea nitrogen concentration after weaning was the lowest in H. Plasma IGF-1 concentrations were higher in H than in L. Ruminal pH was not different, but the molar proportion of ruminal propionate was higher in H than in L throughout the study period. In summary, a high-starch starter may increase propionate production and microbial protein synthesis in the rumen, inducing higher plasma IGF-1 concentrations.

Death-and-culling rates of calves and associated economic losses in the first month of life on dairy farms in eastern Hokkaido, Japan.

The objective of this study was to analyze death and culling (DC) of calves during the first month of life and associated economic losses on dairy farms in eastern Hokkaido, Japan. The DC in the first month of life of 4411 Holstein and Wagyu crossbred calves born in the year 2019-2020 on 39 dairy farms milking Holsteins was investigated. Based on a target DC rate of 6.75%, farms were classified into two groups, those with high DC rates (HDC, 11.68%, n = 18) and those with low DC rates (LDC, 2.67%, n = 21), and analyzed for DC factors (breed, sex, parity of dams, and housing type of dams) and diseases causing DC, their loss estimates, and replenishment of DC calves (birth rate, purchase of heifers, and housing type of dams). Comparisons between groups were made using the Kaplan-Meier method, the Mann-Whitney U test, and chi-square test. The DC rate of Holsteins was significantly higher (P < 0.001) for HDC farms than for LDC ones. But, the DC rate of Wagyu crossbreds was not different between the groups, which suggested hybrid vigor. The DC rates for digestive diseases were significantly higher (P < 0.01) in the HDC farms, except for congenital diseases and deaths of unknown cause. The overall loss estimates of DC per calf-month for all farms was 8892, JPY/calf-months, 14,726 for HDC farms, and 4065 for LDC farms. The loss estimates of items with significant differences in DC rates were significantly higher (P < 0.05) in the HDC farms, with higher loss estimates for Holsteins and digestive diseases being the most common characteristics of HDC farms. In binomial logistic regression analysis with "HDC farms or not" as the response variable and replenishment of DC calves as the explanatory variable, HDC was significantly more likely (OR: 1.10, P < 0.05) on farms with a higher birth proportion of Holsteins, and HDC farms supplemented the DC calves by increasing the birth proportion of Holsteins.

Effects of starch content of calf starter on feed intake, growth performance, and fecal properties in dairy calves under a high plane of milk replacer feeding.

The objective of this study was to investigate the effects of different starch contents in calf starter on the growth, feed intake, and fecal characteristics in dairy calves fed under a high plane of nutrition. Twenty-seven Holstein calves were assigned equally to one of three calf starter treatment groups in a randomized block design: high (H) starch (41.8%); medium (M) starch (31.9%); and low (L) starch (22.0%). Milk replacer was fed up to 1.2 kg/day as fed, gradually reduced from 6 weeks of age; calves were weaned at 8 weeks of age. Each starter (up to 3.5 kg/day as fed) and chopped hay were provided ad libitum from 5 days of age. Fecal samples, along with body measurements, were collected from 4 to 13 weeks of age. The average daily gain after weaning was greater in the H group than in the L group. The average starter intake for 3 days before weaning was not different among treatments but was greater after weaning in the H group than in the L group. Starter starch content did not affect the number of days with diarrhea or fecal starch concentration. The high-starch starter had no adverse effects during the weaning transition.

Epidemiological Features of Postpartum Subclinical Ketosis in Dairy Herds in Hokkaido, Japan.

This study was carried out as an observational study in order to determine the prevalence of postpartum subclinical ketosis (SCK) in dairy herds in Hokkaido, Japan. From April 2012 to March 2014, blood ß-hydroxybutyrate (BHBA) concentration was measured once within 3-88 days in milk (DIM) in 1394 apparently healthy cows from 108 farms to diagnose SCK (≥1.2 mM). In cows within 14 DIM, this was classified as SCK II, and from 15 DIM, this was classified as SCK I. Herds with a combined percentage of SCK I and SCK II of less than 10% were classified as SCK-negative herds, those with percentages of 10-25%, were classified as alert herds, and those with one of 25% or more, we classified as positive herds. The prevalence of SCK in the entire DIM was 17.6%. The prevalence of SCK II (20.2%) tended to occur more frequently than SCK I (16.5%, p = 0.094). The frequency of SCK I was higher at the fourth parity. The number of milking cows in SCK-positive herds was significantly smaller than those of the other two types of herds (p = 0.004). The frequency of SCK-positive herds in tie stalls and with component feeding was higher than for free stall or free barn and with total mixed ration (p = 0.054 and p = 0.002). This study reveals the prevalence of SCK in Hokkaido, Japan, and shows that SCK is associated with parity and the management system.

Sodium butyrate administration modulates the ruminal villus height, inflammation-related gene expression, and plasma hormones concentration in dry cows fed a high-fiber diet.

The objectives of this study were to evaluate the effects of sodium butyrate on the ruminal villus morphology, mRNA expression associated with nutrient metabolism and inflammation in the ruminal epithelium, and plasma concentrations of metabolites and hormones in non-lactating cows fed a high-fiber diet. Four Holstein cows with a rumen cannula were assigned to two treatments in a crossover design. The treatments were ruminal administration of sodium butyrate premix or control premix before feeding to cows fed the same total mixed ration mainly composed of glass silage once a day. Sodium butyrate was provided at a butyrate dose of 0.04% per kg body weight. The control premix was made by replacing sodium-butyrate with wheat bran. The plasma ß-hydroxybutyrate concentration increased 3 to 6 h after the butyrate premix administration but returned to a concentration similar to that of the control before feeding. After continuous administration, increases in the ruminal villus height and plasma concentration of glucagon-like peptide-2, and lower gene expression of TNF-α, IL-1ß, and TLR-2 in the rumen epithelium were observed in cows supplied with the butyrate premix. These results showed that sodium butyrate affects rumen epithelial morphology and plasma concentrations of hormones even under a low fermentable diet.

Performance evaluation of a newly designed on-farm blood testing system for determining blood non-esterified fatty acid and ß-hydroxybutyrate concentrations in dairy cows.

The objective of this study was to evaluate a newly designed on-farm blood testing system (OFBTS) for monitoring blood concentrations of non-esterified fatty acids (NEFA) and ß-hydroxybutyrate (BHBA) in dairy cows. Blood samples from 230 Holstein dairy cows between -86 and 343 days in milk were collected. A drop of whole blood was used to determine NEFA and BHBA using the OFBTS. Plasma from the remaining blood was used to determine both analytes using a commercial kit (gold standard). In the repeatability of the OFBTS, the intra-assay CV for NEFA and BHBA were 1.3% and 4.5%, and the inter-assay CV were 1.8% and 2.9%, respectively. The slope and coefficient of determination of OFBTS analysis of NEFA compared to the gold standard were 0.92 and 0.94. Those for BHBA were 0.94 and 0.98. Mean of the difference between the gold standard laboratory assays and OFBTS of NEFA and BHBA were 0.021 and 0.019, respectively. However, the bias became substantial for NEFA in the higher concentration ranges (>1.2 mEq/L). The sensitivity and specificity of NEFA were 93.2% and 99.4% at a cutpoint of 0.4 mEq/L, and 87.9% and 100% at 0.6 mEq/L. Those of BHBA were 86.2% and 99.0% at a cutpoint of 1.0 mM, and 94.7% and 99.5% at 1.2 mM. The reaction time for the NEFA to reach 0.6 mEq/L was 7 min. The BHBA reaction reached 1.2 mM within 2 min. In conclusion, the OFBTS has excellent performance for evaluating blood NEFA or BHBA concentrations.

The effects of glutathione ethyl ester in in vitro maturation on the developmental ability of oocytes derived from cattle with liver abnormalities.

The main objectives of this study was to identify the effects of a relationship of hyper-concentration of Gamma-glutamyltransferase (γ-GTP) in follicle fluid (FF) on the levels of glutathione (GSH)/reactive oxygen species (ROS) in oocytes and subsequent embryo development in cattle with abnormal livers. Furthermore, we investigated the effect of supplementing in vitro maturation medium with glutathione ethyl ester (GSH-OEt) on the subsequent developmental potential of oocytes from such cattle. We used a control group of cattle (with normal livers) and a liver disorder (LD) group, in which the liver was diagnosed as being abnormal. In experiment 1, the LD group was divided to two subgroups according to the concentration of γ-GTP in FF: a low group (≤50 IU/L; the low LD group), and a high group (>50 IU/L: the high LD group). Cumulus oocyte-complexes (COCs) were matured and fertilized in vitro and then cultured to the blastocyst stage. The levels of GSH and ROS in the matured oocytes after IVM were then assessed in each group. On day 7 after fertilization, embryo cleavage and development were assessed. We found that the rate of development to the blastocyst stage was significantly lower in the high LD group than in the control group and the low LD group. The levels of GSH in matured oocytes were significantly lower in the high LD group than in the control group and low LD group. The levels of ROS in matured oocytes was significantly higher in the high LD group than in the control group and the low LD group. In experiment 2, COCs from cattle in the high LD group were matured in m-199 supplemented with 5 mM GSH-OEt, then IVF and IVC was performed for 7 days. The GSH levels were determined in some COCs after IVM. The supplementation of media with GSH-OEt during IVM increased the levels of GSH in mature oocytes and improved the rate of blastocyst development compared with the control group. In conclusion, GSH-OEt supplementation to media during IVM can improve the developmental potential of oocytes in liver-diseased cattle with high γ-GTP concentrations in the FF by increasing intracellular GSH synthesis and scavenging ROS.

The effect of sodium butyrate supplementation on ruminal and fecal pH and serum lipopolysaccharide-binding protein after ruminal acidosis challenge in nonlactating cows.

The objective of this study was to evaluate effects of sodium-butyrate supplementation on gastrointestinal function and the inflammatory response to ruminal acidosis (RA) challenge in cows. Four nonlactating cows with a rumen cannula were assigned to two treatments in a crossover design. Treatments were ruminal administration of sodium-butyrate (BUT) or control (CON). Sodium-butyrate was provided as Gustor BP70 and administered at a butyrate dose of 0.04% per kg body weight. The CON premix was made by replacing sodium-butyrate with wheat bran. Experimental periods were 28 days long with 21-day washout period separating the treatments. On Day 25 of each period, corn starch was ruminally administered at 0.7% per kg body weight as RA challenge. After RA challenge, ruminal pH was lower, and endotoxin concentration was higher for cows provided with BUT than those with CON, but the increase in fecal starch and the decrease in fecal pH were attenuated by BUT. The effect of butyrate supplementation on serum lipopolysaccharide-binding protein after RA challenge was not found. From these findings, butyrate supplementation mitigated rectal acidosis by reducing the flux of fermentable carbohydrate into the large intestine. An anti-inflammatory effect of butyrate was not observed, possibly due to lower pH and higher endotoxin concentration in the rumen.

Serum paraoxonase-1 activity in tail and mammary veins of ketotic dairy cows.

The objective of this study was to evaluate the association between ketonemia and serum paraoxonase-1 (PON1), malondialdehyde (MDA), and other blood components in tail and mammary veins of dairy cows. Forty-two Holstein dairy cows with decreased feed intake were divided into HIGH (≥ 1.2 mM; n = 31) and LOW (< 1.2 mM; n = 11) groups based on the ß-hydroxybutyrate concentration in plasma collected from the tail vein. The HIGH group had a significantly greater plasma non-esterified fatty acid (NEFA) concentration, but significantly lower serum PON1 activity and phospholipid concentration, and a tendency to have a lower cholesterol ester concentration than the LOW group. Serum PON1 activity was not correlated with the MDA concentration but was positively correlated with serum concentrations of cholesterol esters and phospholipids, and negatively correlated with the plasma NEFA concentration. These results suggest that serum PON1 activity is reduced by hyperketonemia and the relevance of PON1 to MDA seems to not be direct, though it is involved.

L'objectif de la présente étude était d'évaluer l'association entre l'acétonémie et la paraoxonase-1 (PON1), le malondialdéhyde (MDA), et d'autres composés du sang dans les veines caudale et mammaire de vaches laitières. Quarante-deux vaches laitières de race Holstein présentant une diminution de l'ingestion d'aliments furent divisées en groupes ÉLEVÉ (≥ 1,2 mM; n = 31) et BAS (< 1,2 mM; n = 11) basés sur la concentration de ß-hydroxybutyrate de plasma prélevé de la veine caudale. Le groupe ÉLEVÉ avait une concentration plasmatique significativement plus grande d'acides gras non-estérifiés (NEFA), mais le sérum présentait une activité PON1 et une concentration de phospholipides significativement réduite, et une tendance à avoir une concentration d'esters de cholestérol plus faible que le groupe BAS. L'activité de PON1 sérique n'était pas corrélée avec la concentration de MDA mais était corrélée positivement avec les concentrations sériques d'esters de cholestérol et de phospholipides, et corrélée négativement avec la concentration plasmatique de NEFA. Ces résultats suggèrent que l'activité de PON1 sérique est réduite par l'hypercétonémie et la pertinence de PON1 envers MDA ne semble pas être directe, bien qu'elle semble impliquée.(Traduit par Docteur Serge Messier).

Duodenal infusion of fatty acids differentially affects plasma glucagon-like peptide-1 and ghrelin concentrations in sheep.

The aim of this study was to investigate how intraduodenal infusions of fatty acids (FA) affect appetite-related gut peptides such as glucagon-like peptide-1 (GLP-1) and ghrelin in sheep. We hypothesized that these peptides can be highly reactive to unsaturated long-chain FA, because they are well known to decrease dry matter intake (DMI). Four ewes were fitted with a duodenal cannula and a jugular vein catheter for a 6-h duodenal infusion of the 9 FA (C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C18:1, C18:2, and C18:3) and water (control). The concentration of each FA was 1.6 g per metabolic body weight (BW), approximately corresponding to the amount of supplemented fat in a standard dairy cow diet. Each infusion was separated by at least 2 d. During the infusion period, blood samples were collected periodically to determine changes in plasma GLP-1, ghrelin, and metabolite concentrations. Duodenal infusions of C18:1, C18:2, and C18:3 led to higher plasma GLP-1 (P < 0.05) and lower glucose (P < 0.05) than control. Plasma ghrelin concentrations were greater in C18:1 and C18:3 infusions than control (P < 0.05). Plasma ketone bodies were higher in C8:0 and C10:0 infusions (P < 0.05), but plasma triglyceride concentrations were lower in C8:0, C10:0, C12:0, and C16:0 infusions (P < 0.05) than control. Fatty acid infusions except for C18:3 led to higher plasma NEFA concentrations than control (P < 0.05). These results confirmed that the hypophagic effect of dietary unsaturated long-chain FA is mediated by GLP-1 (an anorexigenic effect) secretion. However, we also observed higher plasma ghrelin (an orexigenic effect) partially by unsaturated long-chain FA. Thus, the gut peptide secretions when ruminant animals ingest FA supplements would complexly affect satiety and further studies are needed to determine their each impact on DMI.

Histochemical properties of bovine and ovine mammary glands during fetal development.

In order to obtain more information on the development of bovine and ovine fetal mammary glands, a series of mammary glands from fetuses of different ages were analyzed. A total of 16 bovine fetuses with curved crown rump lengths ranging from 12 cm (80 days) to 75 cm (240 days) and 15 ovine fetuses ranging from 55 days to 131 days were examined. We used hematoxylin and eosin stain and Oil-Red-O stain to analyze the developmental and morphogenetic processes of mammary glands. In addition, we used immunohistochemical staining to determine the pattern of expression of cytokeratin 18 (CK18) during luminal epithelial differentiation, α-smooth-muscle actin (α-SMA) for myoepithelial differentiation, Ki-67 for cell proliferation, and estrogen receptor α (ERα). Our analyzes showed: (a) The primary mammary duct begin to proliferate in a lengthwise within the teat at 90 days in bovine fetuses and 63 days in ovine fetus; (b) luminal epithelial cells and myoepithelial cells appeared from 90 days in bovine fetuses and 63 days in ovine fetus; (c) proliferation of epithelial cells appeared to coincide with the development of the primary and secondary ducts; and (d) ERα was not found in the fetal mammary gland, but adipocytes showed the presence of ERα. Overall, these results indicate that the sequence of events in the prenatal development of the mammary gland of sheep is similar to that of cattle.

Effects of intravenous ghrelin injection on plasma growth hormone, insulin and glucose concentrations in calves at weaning.

Ghrelin action, which stimulates growth hormone (GH) secretion, may alter during the weaning period in calves. Our objective was to compare the effects of intravenous ghrelin injection on plasma GH, insulin and glucose concentrations in calves around the weaning period. Four Holstein bull calves were fed whole milk and allowed free access to solid feeds, and weaned at 7 weeks of age. Measurements were performed at weeks 1, 2, 4, 6, 7, 9, 11 and 13, when calves were intravenously injected with ghrelin (1.0 µg/kg body weight (BW)) through a catheter, and jugular blood samples were obtained temporally relative to the injection time. Estimated digestible energy intake per metabolic BW transiently decreased at week 7 because of low solid intake immediately after weaning, and thereafter gradually increased. Plasma insulin and glucose concentrations were not affected by ghrelin injection at all ages. In contrast, plasma GH concentrations increased with ghrelin injection at all ages. The incremental area of GH at week 7 was greatest and significantly higher compared with weeks 2, 4, 6 and 9. This result suggests that nutrient insufficiency immediately after weaning enhances GH responsiveness to ghrelin.

Comparisons of insulin related parameters in commercial-type chicks: Evidence for insulin resistance in broiler chicks.

The aim of this study is to elucidate whether insulin acts differentially within the central nervous system (CNS) of two types of commercial chicks to control ingestive behavior. Male layer and broiler chicks (4-day-old) were intracerebroventricularly (ICV) injected with saline or insulin under satiated and starved conditions. Feed intake was measured at 30, 60 and 120 min after treatment. Secondly, blood and hypothalamus were collected from both chick types under ad libitum feeding and fasting for 24 h. Plasma insulin concentration was measured by time-resolved fluoro-immunoassay. Hypothalamic insulin receptor mRNA expression levels were measured by quantitative RT-PCR. The ICV injection of insulin significantly inhibited feed consumption in layer chicks when compared with saline (P<0.05), but not broiler chicks (P>0.1). Plasma insulin concentration of both chick types significantly decreased following 24 h of fasting, while insulin concentrations in the broiler chicks were significantly higher compared to the layers fed under ad libitum conditions. Hypothalamic insulin receptor mRNA expression levels were significantly lower (P<0.05) in broiler chicks than in layer ones under ad libitum feeding. Feed deprivation significantly decreased insulin receptor mRNA levels in layer chicks (P<0.01), but not in broiler chicks (P>0.1). Moreover, plasma insulin concentrations correlated negatively with hypothalamic insulin receptor protein expression in the two types of chicks fed ad libitum (P<0.05). These results suggest that insulin resistance exists in the CNS of broiler chicks, possibly due to persistent hyperinsulinemia, which results in a down-regulation of CNS insulin receptor expression compared to that in layer chicks.