Intraruminal infusion of n-butyric acid induces an increase of ruminal papillae size independent of IGF-1 system in castrated bulls

Abstract

The objective of this study was to explore morphological alterations of rumen papillae induced by n-butyric acid in relation to the insulin-like growth factor (IGF) system in adult castrated bulls. Three animals fitted with rumen cannula were fed twice daily at a low and high nutritional level (LL and HL), i.e., at 1.1 × maintenance (M) and 1.6 × M, respectively. Diets contained artificial dried grass and concentrate (74:26 and 52:48). Bulls received no (B0) or daily intraruminal infusions of 500 g n-butyric acid (B500) over 14 d. The infusion started 1 h after the morning feeding (9:00) and lasted for 3.5 h. Thus, four treatments (B0LL, B500LL, B0HL, and B500HL) were compared. Blood and rumen mucosa samples from the atrium ruminis were taken at the last day of each period. Length, width and surface of rumen papillae were greater (p < 0.001) in B0HL than in B0LL. Treatment with n-butyric acid resulted in an increase of the papillae surface of 20 – 40% (p = 0.047) for both nutritional levels as compared to periods without n-butyric acid treatments. The higher nutritional level and intraruminal n-butyric acid infusion induced epithelial cell death. The percentage of proliferative cells was doubled by n-butyric acid treatment. The mRNA of IGF-1 and IGF type 1 receptor (IGF-1R), as well as IGF-1R binding capacity were unaffected by butyric acid treatments. The abundance of IGF-1 mRNA tended to be lower (p = 0.1) and IGF-1R abundance was lower (p = 0.03) in response to the HL. The plasma IGF-1 concentration was lower with butyric acid treatment (p < 0.01), but was unaffected by the nutritional level. In conclusion, under described experimental preconditions of daily short-time intraruminal n-butyric acid infusion alterations of rumen papillae morphology is not mediated by ruminal IGF type 1 receptor and by local IGF-1 expression in papillae in castrated bulls.     

Plasma LH and testosterone in Zebu crossbred bulls after exposure to an estrous cow and injection of synthetic GnRH

Plasma hormone levels were examined in 4 mature Zebu bulls of normal libido (HL) and 4 which were sexually inactive (LL). When used in an artificial insemination programme the 8 bulls had similar fertility. Basal levels of LH and testosterone (T) estimated from 8 sequential blood samples at 30 minute intervals were not different in HL and LL bulls. Exposure of the animals to an estrous cow did not stimulate LH release. Following sexual stimulation plasma T levels actually decreased by an average (±S.E) of 2.9 (±1.9) ng/ml in the HL group and increased by 3.9 (±1.6) ng/ml in the LL group. An injection of 1 mg GnRH (Hoechst) caused LH release of similar magnitude in HL and LL bulls. The elevation of plasma T which followed GnRH injection was significantly larger in HL bulls.Low libido was not associated with a deficiency of basal LH or T, nor with the ability of the pituitary to respond to GnRH.     

Influence of fumaric acid on ruminal parameters and organ weights of growing bulls fed with grass or maize silage

The influence of the potential methane reducer, fumaric acid (FA), on ruminal parameters, the rumen wall and organ weights was investigated in a long-term study with growing bulls. In all, 20 bulls were fed with maize or grass silage as roughage, and with concentrate with or without 300 g FA per animal and day during the whole fattening period. After slaughtering, the organs were weighed and blood serum was analysed for glucose, β-hydroxybutyric acid (BHB) and non-esterified fatty acid concentration. The ruminal fluid was analysed for short-chain fatty acids, ammonia-N and the microbial community via single strand conformation polymorphism analysis. The rumen wall was examined histopathologically and results were graded as ‘no visible lesions’, ‘few inflammatory infiltrates’, ‘some inflammatory infiltrates’ or ‘several inflammatory infiltrates’. In addition, the dimensions of the rumen villi were measured. The FA supplementation decreased the serum BHB concentration and the butyric acid concentration in the ruminal fluid. The microbial community in the ruminal fluid was not influenced by FA. An interaction between FA and silage type was observed for the inflammation centres counted in the villous area of rumen papillae. This interaction was also observed in the length and surface of the rumen villi. Rumen villi results show that the influence of FA depends on the roughage used in the diet.     

Growth hormone and insulin-like growth factor I concentrations in bulls of various growth hormone genotypes

A leucine/valine substitution at amino acid position 127 was identified by the polymerase chain reaction and restriction fragment length polymorphism in the bovine growth hormone gene. Genotyping was performed in 84 AI bulls of three different breeds, in which plasma concentrations of growth hormone (GH) and insulin-like growth factor I (IGF-1) were also measured. Gene frequencies of variants L (leucine) und V (valine) were 0.80/0.20 (Black and White), 0.90/0.10 (Brown), 0.71/0.29 (Simmental). Hormone concentrations were measured during different physiological conditions (normal feeding, fasting, realimentation) in the majority of animals. Generally, genotype LL was associated with higher concentrations of GH than LV. This difference was significant in Black and White bulls (P < 0.05). In contrast, IGF-1 concentrations were higher in LV than in LL animals. This was most pronounced in mature, realimented Simmental bulls. We conclude that the various GH alleles influence the circulating concentrations of GH and IGF-1.     

Surface enlargement in the rumen of free-ranging muskoxen (<Emphasis Type="Italic">Ovibos moschatus</Emphasis>)

The intraruminal papillation pattern indicates the degree of rumen contents stratification and is related to the feeding niche of a ruminant. Muskoxen (Ovibos moschatus) display a variety of morphophysiological adaptations typical for grazers. We investigated the intraruminal papillation of 22 free-ranging muskoxen from five different months by comparing the surface enlargement factor both between seasons and between individual rumen regions. The seasonal pattern of rumen papillation indicated a distinct seasonality in food quality. The intraruminal papillation indicated a moderate degree of rumen contents stratification typical for intermediate feeders. The nutritional ecology of muskoxen is characterised by specific morphophysiological adaptations to a grass-dominated diet that nevertheless allow extensive seasonal use of browse forage.     

Photosynthesis but not CAM responded flexibly to changes in irradiance in Plectranthus marrubioides (Lamiaceae)

Well-watered plants of Plectranthus marrubioides Benth., a crassulacean acid metabolism (CAM) species naturally inhabiting sun exposed succulent places, were grown at photosynthetically active photon flux densities (PPFD) of either 150 (LL) or 300 (HL) μmol m-2 s-1 in a controlled environment. Photosynthesis of LL plants was saturated by irradiance of ca. 500 μmol m-2 s-1 while in HL plants saturation was not reached up to 1200 μmol m-2 s-1 and photosynthetic capacity was nearly 50 % higher than in the LL plants. However, maximum photon yield was 55 % lower and compensation irradiance was 25 % higher in LL plants. The former also had larger, more succulent leaves, i.e., they were morphologically more sun adapted. On the other hand, nocturnal accumulation of malic and citric acid, nighttime CO2 gain, and the low relative carbon recycling were independent of the prevailing PPFD. Furthermore, photosynthetic performance was flexibly and reversibly adjusted in HL plants after transfer to 600 or 150 μmol m-2 s-1 while nocturnal CO2 uptake was not influenced. Photosynthesis showed a high acclimation potential to high PPFD and patterns of gas exchange became more C3-like the higher the irradiance was, without a direct effect on CAM in P. marrubioides.     

Elevated urinary excretion of orotic acid in sheep caused by intraruminal infusion of sodium propionate.

1. The effect of sodium propionate on urinary excretion of orotic acid was investigated. 2. Solutions containing sodium propionate or NaCl, 750 mM/day each, were continuously infused into the rumen for 10 days. 3. During NaCl infusion, an urinary orotic acid excretion of 290 +/- 80 micrograms/day was noted. The intraruminal infusion of sodium propionate raised the concentration of propionic acid in the rumen fluid from 14.0 +/- 0.9 to 26.9 +/- 1.9 mM. 4. During this experimental period the excretion of orotic acid via urine significantly increased to 492 +/- 30 micrograms/day. Parameters of nitrogen balance were not altered by propionate. 5. It is suggested that the site of propionate action in intact sheep is in the pyrimidine synthesis pathway.     

Effect in the cow of intraruminal infusions of volatile fatty acids and of lactic acid on the secretion of the component fatty acids of the milk fat and on the composition of blood

1. The effects in the cow of intraruminal infusions of acetic acid, propionic acid or butyric acid on the secretion of the component fatty acids of the milk fat, and of these acids and of lactic acid on the composition of the blood plasma of the jugular vein, have been studied. 2. The infusion of acetic acid or butyric acid increased the yield of the C₄–C₁₆ acids of milk fat but decreased the yield of C₁₈ acids. The infusion of propionic acid decreased the yields of all major component acids except palmitic acid and possibly lauric acid. 3. The changes in the concentrations in blood plasma of glucose and of ketone bodies were consistent with the glucogenic effect of propionic acid and the ketogenic effects of butyric acid and acetic acid. The effects of lactic acid were not consistent from cow to cow. Only with the infusion of acetic acid was a significant increase in the concentration of total volatile fatty acids in blood plasma found. Infusions of butyric acid and of propionic acid tended to depress the concentration of citric acid in the blood plasma and infusion of acetic acid increased it. No consistent effects of the infused acids on the concentration in blood plasma of esterified cholesterol, free cholesterol, triglyceride or phospholipid were observed. 4. The possibility is discussed that the effects of the infused acids on milk-fat secretion are caused through an alteration of the concentrations of precursors of milk fat in mammary arterial blood.     

Inactivation of enterohemorrhagic Escherichia coli in rumen content- or feces-contaminated drinking water for cattle

Cattle drinking water is a source of on-farm Escherichia coli O157:H7 transmission. The antimicrobial activities of disinfectants to control E. coli O157:H7 in on-farm drinking water are frequently neutralized by the presence of rumen content and manure that generally contaminate the drinking water. Different chemical treatments, including lactic acid, acidic calcium sulfate, chlorine, chlorine dioxide, hydrogen peroxide, caprylic acid, ozone, butyric acid, sodium benzoate, and competing E. coli, were tested individually or in combination for inactivation of E. coli O157:H7 in the presence of rumen content. Chlorine (5 ppm), ozone (22 to 24 ppm at 5 degrees C), and competing E. coli treatment of water had minimal effects (<1 log CFU/ml reduction) on killing E. coli O157:H7 in the presence of rumen content at water-to-rumen content ratios of 50:1 (vol/wt) and lower. Four chemical-treatment combinations, including (i) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.05% caprylic acid (treatment A); (ii) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.1% sodium benzoate (treatment B); (iii) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.5% butyric acid (treatment C); and (iv) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 100 ppm chlorine dioxide (treatment D); were highly effective (>3 log CFU/ml reduction) at 21 degrees C in killing E. coli O157:H7, O26:H11, and O111:NM in water heavily contaminated with rumen content (10:1 water/rumen content ratio [vol/wt]) or feces (20:1 water/feces ratio [vol/wt]). Among them, treatments A, B, and C killed >5 log CFU E. coli O157:H7, O26:H11, and O111:NM/ml within 30 min in water containing rumen content or feces, whereas treatment D inactivated approximately 3 to 4 log CFU/ml under the same conditions. Cattle given water containing treatment A or C or untreated water (control) ad libitum for two 7-day periods drank 15.2, 13.8, and 30.3 liters/day, respectively, and cattle given water containing 0.1% lactic acid plus 0.9% acidic calcium sulfate (pH 2.1) drank 18.6 liters/day. The amounts of water consumed for all water treatments were significantly different from that for the control, but there were no significant differences among the water treatments. Such treatments may best be applied periodically to drinking water troughs and then flushed, rather than being added continuously, to avoid reduced water consumption by cattle.     

Sulfur and methionine metabolism in sheep. I. First approximations of sulfur pools in and sulfur flows from the reticulo-rumen.

Sulfur pools in the rumen and sulfur flows from the rumen were investigated in two experiments with sheep on a diet containing equal parts of oaten and lucerne chaffs. The diet was fed at two levels, either chopped or pelleted, and with intraruminal DL-methionine supplements. Ruminal fluid volumes and fluid flows to the omasum were measured. None of the treatments influenced ruminal fluid volume. Fluid flow to the omasum, however, was increased by increasing dry matter intake (DMI), and was further enhanced by feeding chaffed hay rather than the same materials ground and pelleted; the DL-methionine supplement had no effect. First approximation of the ruminal sulfur pools and of sulfur flows to the omasum were derived from the concentration of sulfur in true digesta and the ruminal fluid volume or fluid flow. Increasing DMI from 500 to 1000 g/day resulted in larger ruminal pools of total (1096 v. 792 mg), neutral (1016 v. 731 mg) and protein (479 v. 419 mg) sulfur, but the reducible sulfur pools were not affected by the level of DMI. Infusions of DL-methionine increased the ruminal sulfide sulfur pool irrespective of level of DMI. The first approximation of total sulfur flow was increased by 1660 mg/day at the higher level of DMI, due mainly to increases of 710 mg S/day as protein sulfur and 859 mg S/day as non-protein neutral sulfur. Flows of inorganic sulfate and ester sulfate sulfur, although small in comparison with organic sulfur flows, increased with level of DMI. Sulfide sulfur flows were also increased at the higher level of DMI, and were almost doubled by intraruminal infusions of DL-methionine.     

Influence of ruminal and plasma osmotic pressure on salivary secretion in sheep

The osmotic pressure of the rumen contents and also of the blood was altered by intraruminal administration of water or hypertonic solutions. It was found that alterations in osmotic pressure were accompanied by inverse changes in the flow rate of mixed saliva. Intravenous infusion of hypertonic solutions, causing elevation of the osmotic pressure of the blood without affecting that of the rumen, also caused a reduction of salivary secretion rate. The flow rates of both parotid and residual saliva were affected. When strongly hypertonic solutions of sodium or potassium salts were infused into the rumen, or sodium salts or urea were infused into the blood, the concentration of those substances increased in the saliva. Other treatments had little effect on salivary composition.     

Kinetics of ethanol metabolism in sheep.

Kinetic aspects of ethanol metabolism were studied in sheep after intravenous or intraruminal infusion of ethanol. Vmax and Km in fed animals were respectively 295 +/- 10 mg.h-1.l-1 (l = litre of body water) and 32.1 +/- 2.4 mg.l-1. Elimination half-life was 1.47 +/- 0.26 h. The corresponding values in the fasted animal were not significantly different. During venous infusion an increase in plasma acetate, inversely correlated to plasma ethanol, was observed. No modification in glycemia occurred. Intraruminal infusion of ethanol increased the concentration of all SCFA in the rumen juice, the largest part of this modification being relative to acetate. Repetition of the infusion over a period of 11 consecutive days increased the number of SCFA in the rumen, indicating microflora adaptation to ethanol utilization. Taking into account the range of ethanol concentrations found in silage (10-50 g.kg-1 BW) we can consider that ethanol is readily metabolized simultaneously by the rumen microflora and the enzymatic system of the host. With a corresponding daily intake of ethanol (0.2-1 g.kg-1 BW) both systems are not saturated and plasma ethanol level always remains below 0.25 g.l-1.     

Effect of starter diet supplementation on rumen epithelial morphology and expression of genes involved in cell proliferation and metabolism in pre-weaned lambs

Starter feeding is usually used in lamb production to improve rumen development and to facilitate the weaning process, but molecular mechanism of which is not well understood. Therefore, the objective of this study is to investigate the effect of starter feeding on the expression of ruminal epithelial genes involved in cell proliferation, apoptosis and metabolism in pre-weaned lambs. We selected eight pairs of 10-day-old lamb twins. One twin was fed ewe milk (M, n=8), while the other was fed ewe milk plus starter (M+S, n=8). The lambs were sacrificed at 56 days age. Results showed that the lambs fed M+S had lower pH in the rumen and a higher concentration of acetate, propionate, butyrate and total volatile fatty acid (VFA). Compared with the M group, the concentration of β-hydroxybutyric acid in plasma had an increased trend, and the concentration of IGF-1 in plasma had an decreased trend in the M+S group. The length, width and surface of rumen papillae increased in the M+S group compared with the M group; this was associated with increased cell layers in the stratum corneum, stratum granulosum and total epithelia. Messenger RNA (mRNA) expression of proliferative genes of cyclin A, cyclin D1 and cyclin-dependent kinase 2 in the ruminal epithelia of M+S lambs was increased compared with M only lambs. The mRNA expression of apoptosis genes of caspase-3, caspase-8, B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in the M+S group was decreased compared with M group, but the ratio of Bcl-2 to Bax were not changed between the two groups. Expression of IGF-1 mRNA was decreased, but the mRNA expression of IGF-1 receptor was higher in ruminal epithelia in the M+S group. Furthermore, the mRNA expression of VFA absorption and metabolism genes of β-hydroxybutyrate dehydrogenase isoforms 1 and 3-hydroxy-3-methylglutaryl-CoA lyase had an increased trend in the M+S group than in the M group, but the mRNA expression of 3-hydroxy-3-methylglutaryl-CoA synthase isoform 1, monocarboxylate transporter isoform 1 and putative anion transporter isoform 1 had a decreased trend in the M+S group than in the M group. These results suggest that starter feeding increased proliferation and inhibited apoptosis of ruminal epithelial cells, and may promote the VFA metabolism in ruminal epithelium in pre-weaned lambs. These findings provide new insights into improving rumen development by nutritional intervention strategies in pre-weaned lambs.     

Inactivation of Enterohemorrhagic Escherichia coli in Rumen Content- or Feces-Contaminated Drinking Water for Cattle

Cattle drinking water is a source of on-farm Escherichia coli O157:H7 transmission. The antimicrobial activities of disinfectants to control E. coli O157:H7 in on-farm drinking water are frequently neutralized by the presence of rumen content and manure that generally contaminate the drinking water. Different chemical treatments, including lactic acid, acidic calcium sulfate, chlorine, chlorine dioxide, hydrogen peroxide, caprylic acid, ozone, butyric acid, sodium benzoate, and competing E. coli, were tested individually or in combination for inactivation of E. coli O157:H7 in the presence of rumen content. Chlorine (5 ppm), ozone (22 to 24 ppm at 5°C), and competing E. coli treatment of water had minimal effects (<1 log CFU/ml reduction) on killing E. coli O157:H7 in the presence of rumen content at water-to-rumen content ratios of 50:1 (vol/wt) and lower. Four chemical-treatment combinations, including (i) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.05% caprylic acid (treatment A); (ii) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.1% sodium benzoate (treatment B); (iii) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.5% butyric acid (treatment C); and (iv) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 100 ppm chlorine dioxide (treatment D); were highly effective (>3 log CFU/ml reduction) at 21°C in killing E. coli O157:H7, O26:H11, and O111:NM in water heavily contaminated with rumen content (10:1 water/rumen content ratio [vol/wt]) or feces (20:1 water/feces ratio [vol/wt]). Among them, treatments A, B, and C killed >5 log CFU E. coli O157:H7, O26:H11, and O111:NM/ml within 30 min in water containing rumen content or feces, whereas treatment D inactivated approximately 3 to 4 log CFU/ml under the same conditions. Cattle given water containing treatment A or C or untreated water (control) ad libitum for two 7-day periods drank 15.2, 13.8, and 30.3 liters/day, respectively, and cattle given water containing 0.1% lactic acid plus 0.9% acidic calcium sulfate (pH 2.1) drank 18.6 liters/day. The amounts of water consumed for all water treatments were significantly different from that for the control, but there were no significant differences among the water treatments. Such treatments may best be applied periodically to drinking water troughs and then flushed, rather than being added continuously, to avoid reduced water consumption by cattle.     

Effects of dietary inclusion of Moringa oleifera leaf meal on nutrient digestibility,rumen fermentation,ruminal enzyme activities and growth performance of buffalo calves

This study was conducted to investigate the impact of dietary inclusion of Moringa oleifera leaf meal (MLM) as a substitution for soybean meal on nutrient digestibility, rumen fermentation, rumen enzyme activity, blood metabolites, growth-related hormones, and growth performance of buffalo calves. Thirty buffalo calves eight to nine months of age with an average body weight of approximately 153.7 ± 0.97 kg were randomly distributed through three dietary treatments (ten calves/treatment). MLM inclusion rates were 15% (M15) and 20% (M20), replacing soybean meal by 50 and 75% in the concentrate mixture, respectively. The results indicated that, digestibility of dry matter, organic matter (OM), and crude fiber (CF) increased significantly (p < 0.05) with MLM inclusion, while the digestibility of crude protein (CP) and ether extract (EE) reduced significantly (p < 0.05) with MLM addition. Dietary supplementation with MLM significantly affected (p < 0.001) rumen fermentation by reducing ruminal enzymes, ruminal ammonia-N, total protozoa, and acetate/propionate ratio and increasing acetic, propionic, and butyric acids and total volatile fatty acid concentrations (p < 0.001). Furthermore, dietary inclusion of 15% MLM significantly improved (p < 0.001) final body weight, dry matter intake of feed, daily weight gain, feed conversion efficiency, blood metabolites, and plasma insulin growth factor-I (IGF-I). It can be concluded that MLM is a multi-purpose protein supplement that provides some nutritional and therapeutic advantages when replacing 50% of soybean meal. Dietary supplementation of 15% MLM improved rumen fermentation, growth performance, blood metabolites, plasma IGF-I and mitigated ammonia and methane without any adverse effects in growing buffalo calves.     

Classification of the insulin-like growth factor binding proteins into three distinct categories according to their binding specificities

Competitive binding experiments with insulin-like growth factor (IGF)-1, IGF-2 and des-(1-3)-IGF-1 have confirmed the interpretation based on limited amino-terminal sequence analysis that at least three types of IGF binding protein occur. In addition to the acid stable subunit of the large serum binding protein which exhibits des-(1-3)-IGF-1 binding only slightly less than IGF-1, the small IGF binding proteins can be separated into two classes based on differences in des-(1-3)-IGF-1 and IGF-2 binding potencies.     

Photosynthesis but not CAM responded flexibly to changes in irradiance in Plectranthus marrubioides (Lamiaceae)

Well-watered plants of Plectranthus marrubioides Benth., a crassulacean acid metabolism (CAM) species naturally inhabiting sun exposed succulent places, were grown at photosynthetically active photon flux densities (PPFD) of either 150 (LL) or 300 (HL) μmol m-2 s-1 in a controlled environment. Photosynthesis of LL plants was saturated by irradiance of ca. 500 μmol m-2 s-1 while in HL plants saturation was not reached up to 1200 μmol m-2 s-1 and photosynthetic capacity was nearly 50 % higher than in the LL plants. However, maximum photon yield was 55 % lower and compensation irradiance was 25 % higher in LL plants. The former also had larger, more succulent leaves, i.e., they were morphologically more sun adapted. On the other hand, nocturnal accumulation of malic and citric acid, nighttime CO2 gain, and the low relative carbon recycling were independent of the prevailing PPFD. Furthermore, photosynthetic performance was flexibly and reversibly adjusted in HL plants after transfer to 600 or 150 μmol m-2 s-1 while nocturnal CO2 uptake was not influenced. Photosynthesis showed a high acclimation potential to high PPFD and patterns of gas exchange became more C3-like the higher the irradiance was, without a direct effect on CAM in P. marrubioides. This revised version was published online in June 2006 with corrections to the Cover Date.     

Insulin and Insulin-like Growth Factor-1 Receptors Act as Ligand-specific Amplitude Modulators of a Common Pathway Regulating Gene Transcription

Insulin and insulin-like growth factor-1 (IGF-1) act on highly homologous receptors, yet in vivo elicit distinct effects on metabolism and growth. To investigate how the insulin and IGF-1 receptors exert specificity in their biological responses, we assessed their role in the regulation of gene expression using three experimental paradigms: 1) preadipocytes before and after differentiation into adipocytes that express both receptors, but at different ratios; 2) insulin receptor (IR) or IGF1R knock-out preadipocytes that only express the complimentary receptor; and 3) IR/IGF1R double knock-out (DKO) cells reconstituted with the IR, IGF1R, or both. In wild-type preadipocytes, which express predominantly IGF1R, microarray analysis revealed ∼500 IGF-1 regulated genes (p < 0.05). The largest of these were confirmed by quantitative PCR, which also revealed that insulin produced a similar effect, but with a smaller magnitude of response. After differentiation, when IR levels increase and IGF1R decrease, insulin became the dominant regulator of each of these genes. Measurement of the 50 most highly regulated genes by quantitative PCR did not reveal a single gene regulated uniquely via the IR or IGF1R using cells expressing exclusively IGF-1 or insulin receptors. Insulin and IGF-1 dose responses from 1 to 100 nm in WT, IRKO, IGFRKO, and DKO cells re-expressing IR, IGF1R, or both showed that insulin and IGF-1 produced effects in proportion to the concentration of ligand and the specific receptor on which they act. Thus, IR and IGF1R act as identical portals to the regulation of gene expression, with differences between insulin and IGF-1 effects due to a modulation of the amplitude of the signal created by the specific ligand-receptor interaction.     

Effects of isovalerate supplements on morphology and functional gene expression of rumen mucosa in pre- and post-weaning dairy calves

Isovalerate supplements could stimulate rumen development by improving morphology and function of rumen mucosa, and then promote the growth of calves. This study was done to evaluate the effects of isovalerate supplements on morphology and functional gene expression of rumen mucosa in dairy calves. In total, 48 Chinese Holstein male calves with 15 days of age and 45.1±0.36 kg of BW were randomly assigned to four groups. The treatments were: control, low-isovalerate, moderate-isovalerate and high-isovalerate with 0, 3, 6 and 9 g isovalerate per calf per day, respectively. Supplementary isovalerate was hand-mixed into milk in pre-weaning calves and into concentrate portion in post-weaning calves. The study consisted of a 15-day-adaptation period and a 60-day-sampling period. Calves were weaned at 60 days of age. Three calves were slaughtered from each of the four treatments at 30, 60 and 90 days of age. The weight of body and stomach were measured, samples of ruminal tissues and blood were analyzed. Total stomach weight, total stomach to BW ratio, rumen wall and keratinized layer thickness, serum growth hormone and IGF-1 for both pre- and post-weaning calves increased linearly with increasing isovalerate supplements. Rumen to total stomach weight ratio, the length and width of rumen papillae, and serum β-hydroxybutyrate increased linearly for post-weaning calves. However, abomasum weight to total stomach weight ratio decreased linearly for both pre- and post-weaning calves. The relative messenger RNA expression for growth hormone receptor, IGF-1 receptor and 3-hydroxy-3-methylglutaryl-CoA synthase 1 in rumen mucosa increased linearly for post-weaning calves. Our results suggested that isovalerate supplements promoted rumen development in a dose-dependent manner. The optimum dose was 6.0 g isovalerate per calf per day.