Effect of excess levels of lysine and leucine in wheat‐based,amino acid‐fortified diets on the mRNA expression of two selected cationic amino acid transporters in pigs

An experiment was conducted to evaluate the effect of excess levels of Leu and Lys on the expression of b^0,+ and CAT‐1 mRNA in jejunum, liver and the muscles Longissimus dorsi (LDM) and Semitendinosus (STM). Twenty pigs with an average initial BW of 16.4 ± 1.7 kg were used in a Randomized Complete Block. Dietary treatments (T) were as follows: T1, basal diet; T2, basal plus 3.5 g l ‐Lys/kg diet; T3, basal plus 1.5 g l ‐Leu/kg diet; T4, basal plus 3.5 g l ‐Lys plus 1.5 g l ‐Leu/kg diet. Diets in T1 and T3 met 100% the requirement of Lys for pigs within the 10 to 20 kg body weight range; diets in T2 and T4 contained 35% excess of Lys. Also, diets in T1 and T2 supplied 104%, whereas diets in T3 and T4 supplied 116% the requirement of Leu. The expression of b^0,+ in jejunum was reduced (p = 0.002) because of the supplementation of l ‐Leu, but l ‐Lys supplementation had no effect (p = 0.738). In contrast, the expression of b^0,+ in STM (p = 0.012) and liver (p = 0.095) was reduced by the high level of Lys, but Leu had no effect (p > 0.100). CAT‐1 expression in STM increased by high Lys (p = 0.023) and Leu (p = 0.007) levels. In liver, the expression of CAT‐1 substantially increased (p = 0.001) because of Lys. In conclusion, excess levels of dietary Lys and Leu affect the expression of cationic amino acid transporters, and this effect varies depending on the studied tissue.     

Plasma, tissue, and urine carnitine concentrations in healthy adult cats and kittens

Mean carnitine concentrations [( carnitine]) were higher (P less than 0.05) in adult cats than in kittens for skeletal muscle (total and free carnitine), myocardium (free carnitine), and urine (total and free carnitine). The free/total carnitine ratio was lower (P less than 0.05) in kittens than in adults for liver, myocardium, and urine. Carnitine concentrations were similar between genders in kittens, but in adult cats, [carnitine] in plasma (total, free, and esterified carnitine) and liver (total and free carnitine) were higher (P less than 0.05) in female than in male cats. Total and free plasma [carnitine] were correlated to total and free liver [carnitine], respectively. Skeletal muscle [carnitine] was not correlated to plasma [carnitine]. Correlations in [carnitine] between plasma and myocardium, kidney, or urine were inconsistent.     

l‐carnitine and fat type in the maternal diet during gestation and lactation modify the fatty acid composition and expression of lipid metabolism‐related genes in piglets

This study examined the influence of adding different amounts of maternal dietary l ‐carnitine and two fat types on fatty acid (FA) composition and the expression of lipid metabolism‐related genes in piglets. The experiment was designed as a 2 × 2 factorial with two fat types (3.5% soyabean oil, SO, and 3.5% fish oil, FO) and two levels of l ‐carnitine (0 and 100 mg/kg) added to the sows' diets. A higher proportion of n‐3 polyunsaturated fatty acids (PUFA) and a lower ratio of n‐6/n‐3 PUFA in sow milk and piglet tissues were observed in the FO groups than in the SO groups. Adding l ‐carnitine increased the proportion of C16:1 in sow milk and decreased n‐3 PUFA in piglet subcutaneous fat. Hepatic peroxisome proliferator‐activated receptor α (PPAR‐α) was more abundantly expressed in piglets from the FO groups than from the SO groups (p < 0.05), whereas stearoyl‐CoA‐desaturase (SCD), sterol regulatory element binding protein‐1 (SREBP1) and ?6‐desaturase (D6D) genes were less expressed in the FO groups compared with piglets from the SO groups. The expression of fatty acid synthase (FAS) genes was decreased in the SO groups with l ‐carnitine compared to that of the other dietary treatments. No differences among dietary treatments were observed with regard to the expression of acetyl‐CoA carboxylase (ACC). In conclusion, FO and l ‐carnitine supplementation in sows affect FA composition and hepatic gene expression in piglets.     

Effect of dosage and application mode of l‐carnitine on plasma lipid and egg‐yolk cholesterol of turkeys,hatchability of eggs and post‐hatch growth of their offsprings

The effect of dosage and application mode of l ‐carnitine on plasma lipid and egg‐yolk cholesterol of breeder turkeys, hatchability of eggs and post‐hatch growth response was investigated using 180 breeder hens. The hens were assigned to six dietary treatments in a 2 × 3 factorial arrangements of two application modes of l ‐carnitine (diet and drinking water) supplemented at 0, 50 and 100 ppm (mg/kg or mg/l) levels, respectively. Each treatment was replicated five times with six hens per replicate. Dietary inclusion of 50 ppm l ‐carnitine showed the lowest (p < 0.01) plasma total cholesterol (TC) and low‐density lipoprotein concentration (LDL). Breeder hens offered 50 ppm l ‐carnitine with no regard to application mode recorded the highest (p < 0.01) plasma high‐density lipoprotein (HDL). Hens offered 50 and 100 ppm l ‐carnitine irrespective of application mode also showed reduced (p < 0.01) egg‐yolk TC concentration at 32 weeks of age. Dietary supplementation of 50 ppm l ‐carnitine for breeder turkeys recorded the lowest (p < 0.01) egg‐yolk triglyceride (TG) at 40 weeks of age. Hens offered 50 ppm l ‐carnitine irrespective of application mode recorded the highest (p < 0.05) hen‐day egg production. Incidence of dead‐in‐shell also reduced (p < 0.05) with increasing dosage of l ‐carnitine. Dietary supplementation of 50 ppm and oral application in drinking water of 100 ppm l ‐carnitine for breeder turkeys resulted in highest (p < 0.05) egg fertility. Offsprings from breeder hens fed diets supplemented with l ‐carnitine recorded no post‐hatch mortality. Highest (p < 0.05) post‐hatch final live weight and weight gain was obtained with poults obtained from hens fed diet supplemented with 50 ppm l ‐carnitine. In conclusion, dietary supplementation of 50 ppm l ‐carnitine for turkey hens showed improved serum lipid profile, egg fertility, reduced dead‐in‐shell, egg‐yolk cholesterol and resulted in improved post‐hatch growth performance.     

Impact of dietary manganese concentration on status criteria to determine manganese requirement in piglets

The Mn requirement for pigs is not well established. This study aimed to find criteria for assessing growing piglet supply status for Mn and to determine whether the current Mn recommendations meet the requirements for piglets. Thirty‐six weaned male castrated 27‐day‐old piglets (7.24 ± 0.69 kg) were randomized into six groups of six piglets each and housed individually in stainless steel metabolic cages for 42 days. The piglets were fed a diet based on skimmed milk powder and corn starch with increasing Mn concentrations (0.24; 2; 4; 8; 16; or 32 mg Mn/kg diet as‐fed). In week 6, Mn0.24 led to reduced feed intake (p < 0.05). Manganese concentrations in blood, liver, kidney, lung, heart, phalanx proximalis, pancreas and skeletal muscle were influenced by the dietary Mn supply (p < 0.05). The activity of the Mn‐containing superoxide dismutase in the heart as well as relative arginase activity in the liver were lower in groups Mn0.24, Mn2 and Mn4 compared with the higher supplemented groups (p < 0.05). The relative arginase activity increased clearly with enhanced dietary Mn up to 16 mg/kg and correlated with Mn concentration in the liver. Manganese concentrations in the liver, kidney and phalanx proximalis seem to be suitable biomarkers for Mn status. A 4 mg/kg dietary Mn concentration recommended by NRC (1998, Nutrient Requirements of Swine. National Academy Press, Washington DC.) did not fulfil piglet requirements. Under the conditions investigated, 16 mg Mn/kg diet were necessary to reach a plateau in specific enzyme activity and Mn concentration in organs.     

Development of an ELISA to detect circulating anti‐asparaginase antibodies in dogs with lymphoid neoplasia treated with Escherichia coli l‐asparaginase

Resistance to Escherichia coli l ‐asparaginase in canine lymphoma occurs frequently with repeated administration, a phenomenon often attributed, without substantiation, to the induction of neutralizing antibodies. To test the hypothesis that treated dogs develop antibodies against the drug, we created an enzyme‐linked immunosorbent assay (ELISA) to measure plasma anti‐asparaginase immunoglobulin G responses. Using samples from dogs that had received multiple doses, specific reactivity against l ‐asparaginase was demonstrated, while naïve patients' samples were negative. The optimized ELISA appeared sensitive, with endpoint titers >1 600 000 in positive control dogs. Intra‐ and inter‐assay coefficients of variation were 3.6 and 14.5%. The assay was supported by the observation that ELISA‐positive plasma could immunoprecipitate asparaginase activity. When clinical patients were evaluated, 3/10 dogs developed titers after a single injection; with repeated administration, 4/7 dogs were positive. l ‐asparaginase antibodies showed reduced binding to the PEGylated drug formulation. The ELISA should prove useful in investigating the potential correlation of antibody responses with resistance.     

Effect of L‐theanine on meat quality,muscle amino acid profiles,and antioxidant status of broilers

This study investigated the effect of L ‐theanine on carcass traits, meat quality, muscle antioxidant capacity, and amino acid (AA) profiles of broilers. Three hundred 1‐day‐old Ross 308 male broilers were randomly allotted to five groups with six replicates. Birds were fed the basal diet or basal diet with 300, 600, 900, or 1,500 mg/kg L ‐theanine for 42 consecutive days. The results showed that L ‐theanine quadratically increased dressing percentage, eviscerated percentage, and leg muscle yield (p < .05). Meanwhile, drip loss, cooking loss, shear force, L*_24h, and muscle lactate content decreased quadratically in response to dietary L ‐theanine supplementation (p < .05), while pH_24h and muscle glycogen content were quadratically improved by L ‐theanine (p < .05). Notably, the contents of muscle malondialdehyde and protein carbonyl, and the activities of muscle total antioxidant capacity, catalase, and glutathione peroxidase decreased quadratically in response to dietary L ‐theanine supplementation (p < .05), suggesting that the oxidative stress level of muscle was decreased quadratically. Moreover, L ‐theanine quadratically increased the concentrations of most of muscle essential AA, nonessential AA, and flavor AA (p < .05). In conclusion, L ‐theanine can be used as a valuable feed additive to modulate carcass traits, meat quality, muscle antioxidant status, and AA profiles of boilers, and its optimum addition level is 600 mg/kg based on the present study.     

Tissue and plasma antioxidant status in response to dietary methionine concentration and source in broilers

This study hypothesized that plasma and tissue antioxidant status of broilers is positively influenced when dietary Met concentrations exceed, and negatively when they go below NRC recommendations. In addition, different Met sources are hypothesized to affect the antioxidant defence system differently. Day‐old male Cobb‐500 broilers (n  = 336) were allotted to seven groups and phase‐fed three wheat–soya bean meal‐based basal diets during days 1‐10, 11‐21 and 22‐35. The basal diets (Met? group, Met + Cys concentration 15% below NRC recommendations) were supplemented with 0.10%, 0.25% or 0.40% Met either as DL ‐Met (DLM ) or DL ‐2‐hydroxy‐4‐(methylthio) butanoic acid (DL ‐HMTBA ) (equimolar comparison). Growth performance and carcass weights were lower in the Met? group compared to the groups whose diets met or exceeded Met requirements. The antioxidant defence system was not influenced by the Met source. However, in the liver, concentrations of glutathione increased with increasing dietary Met concentrations. Tocopherol concentrations in the liver at days 10 and 21 were lower in the Met? group than in the groups supplemented with Met. However, liver concentrations of thiobarbituric acid reactive substances (TBA ‐RS ) and protein carbonyls (PC ) were largely not influenced by dietary Met concentration. Plasma tocopherol concentrations at day 35 were lower, and those of TBA ‐RS and PC at day 35 were higher in Met? group than in the groups fed the Met‐supplemented diets. In jejunum, but not in liver, relative mRNA abundances and activities of superoxide dismutase, catalase and glutathione peroxidase were higher in the Met? group than in the groups fed Met‐supplemented diets. These data indicate that suboptimum supply of Met results in decreased antioxidant concentrations in plasma and body tissues, and increases oxidative stress in the jejunum mucosa. However, supplementation of Met in excess of the requirements (based on NRC ) compared to diets adequate in Met + Cys did not influence the antioxidant defence system.     

In ovo injection of branched‐chain amino acids: Embryonic development,hatchability and hatching quality of turkey poults

In this study, the influence of a branched‐chain amino acid blend (BCAA composed of 3 l ‐leucine:1 l ‐valine:2 l ‐isoleucine) injected into the amniotic fluid was evaluated for embryonic growth, yolk‐sac (YS) utilization and development of gastrointestinal tract (GIT) and skeletal muscles of turkey embryos from day 24 of incubation (24E) to hatching, together with hatchability, poult quality and liver L* (lightness), a* (redness) and b* (yellowness) values at hatch. At day 22 of incubation, embryonated eggs (n = 240) were assigned to three treatments, that is, eggs were not injected (control, NC) or injected with 1.5 ml sterile solution with 0.9% salt (SA) or 0.2% BCAA blend (BCAAb). These solutions were injected manually into the amniotic fluid of the embryonated eggs. To determine weights and lengths (where appropriate) of the studied organs and tissues, four embryonated eggs and poults per treatment were selected at 24E and at hatch. While the BCAAb decreased the YS and embryo weight, hatchability and the liver L* value, it increased the weight and quality of poults and the weights of breast and thigh muscles at hatch. In conclusion, the in ovo feeding of the BCAA blend negatively affected hatchability but positively affected hatching weight and poult quality by improving development of skeletal muscles and by regulating energy metabolism.     

Effects of adding liquid dl‐methionine hydroxy analogue‐free acid to drinking water on growth performance and small intestinal morphology of nursery pigs

This study was conducted to evaluate the effect of adding liquid dl ‐methionine hydroxy analogue free acid (LMA) to drinking water on growth performance, small intestinal morphology and volatile fatty acids in the caecum of nursery pigs. Twenty‐four crossbred pigs (Large White × Landrace, BW ～18 kg) were divided into three groups with four replications of two piglets each. The piglets received drinking water without (control), with 0.05 or 0.10% LMA. The results indicated that adding LMA at 0.10% to drinking water significantly increased their weight gain, average daily feed intake (p < 0.05) and tended to improve the feed conversion ratio. Adding LMA to drinking water significantly increased their water intake and significantly reduced the pH of drinking water (p < 0.01), thus total plate count (p < 0.01) and Escherichia coli in drinking water was reduced (p < 0.05), while the total number of bacteria in the caecum was not significantly affected. Liquid dl ‐methionine hydroxy analogue free acid supplementation in drinking water tended to decrease pH in the stomach, duodenum, jejunum, colon and rectum. Furthermore, adding LMA at 0.10% significantly increased villous height in the duodenum, jejunum and ileum (p < 0.05), and the villous height:crypt depth ratio in the jejunum and ileum (p < 0.01) was higher, whereas acetic acid concentration in the caecum was significantly lower than in the control group. It could be concluded that adding LMA to drinking water improved growth performance of the nursery pigs because of high water quality and high nutrient utilization caused by an improvement of small intestinal morphology (not from nutritional effect of methionine source).     

The vitamin E distribution in serum, liver, adipose and muscle tissues in the pig during depletion and repletion

This study was designed to examine the storage capacity for vitamin E in liver, adipose tissue and skeletal muscle of growing pigs during a period of supplementation and of depletion. Therefore, biopsy specimens of these tissues and samples of serum were frequently taken from 7 pigs throughout the experimental period. After a 7-week period on a diet highly supplemented with vitamin E (405 mg/kg), a significant increase (p less than 0.001) in the concentration of this vitamin was observed in all tissues sampled. The highest level (102.9 +/- 26.2 mg/kg) was observed in the liver. After 2 days of depletion the concentration of vitamin E in the liver had fallen by 80%, whereas the concentration in the fat and muscle remained unchanged during 1 week of depletion. The serum vitamin E value rose significantly (p less than 0.001) after 1 week on the supplemented diet and then remained at about 7 mg/l throughout the supplementation period and decreased (p less than 0.001) after 2 days on a nonsupplemented diet. Generally, the serum vitamin E levels among growing pigs are between 2 and 3 mg/l. The results show that the serum and liver values were correlated when the serum level was within this range. Moreover, it is clearly demonstrated that the concentrations of vitamin E in serum and liver reflect the immediate nutritional status of the animal, whereas the vitamin concentrations in adipose and skeletal muscle tissue reflect its long-term nutritional history.     

Application of the slope‐ratio growth assay technique to estimate tryptophan availability in soybean meal fed to young rats

The slope‐ratio assay for rat was used to determine whether tryptophan (Trp) availability in soybean meal (SBM) is affected by the presence of other amino acids (AAs). In a preliminary study, rats were fed graded levels of Trp‐supplemented diets to establish the Trp concentration range over which the weight gain response was linear. This range was found to be from 0.04% to 0.12% Trp. Subsequently, rats were fed basal (0.045% Trp) or Trp‐supplemented diets from three different sources: l ‐Trp alone, SBM, or l ‐Trp mixed with other AAs to reflect AA levels in the test SBM (AA‐mix). Weight gain in rats increased linearly with supplemental Trp intake (p < .05) for all Trp sources. Compared to the slope achieved with l ‐Trp alone, the estimated availability of Trp in SBM was 84.4%, while for the AA‐mix it was 93.4%. It is evident that the 6.6% reduction in l ‐Trp availability in AA‐mix is due to metabolic costs derived from excess levels of other AAs beside Trp, given that the absorption of crystalline l ‐Trp in the small intestine is 100%. In conclusion, the Trp availability of SBM was estimated to be around 90.4% (i.e., 84.4/93.4 × 100) after correcting for the effects of the other AAs in SBM.     

Effects of feeding l-carnitine to gilts through day 70 of gestation on litter traits and the expression of insulin-like growth factor system components and l-carnitine concentration in foetal tissues

We investigated the influence of supplemental l -carnitine on foetal blood metabolites, litter characteristics, l -carnitine concentration in skeletal muscle and insulin-like growth factor (IGF) axis components in foetal hepatic and skeletal muscle tissues at day 40, 55 and 70 of gestating gilts. A total of 59 gilts (body weight = 137.7 kg) received a constant feed allowance of 1.75 kg/day and a top-dress containing either 0 or 50 ppm of l -carnitine starting on the first day of breeding through the allotted gestation length. Foetuses from the gilts fed diets with l -carnitine tended to be heavier (p = 0.06) and the circulating IGF-II tended to be lower (p = 0.09) at day 70, compared with the foetuses from the control gilts. Insulin-like growth factor-I messenger RNA (mRNA) was lower (p = 0.05) in hepatic tissue in the foetuses collected from gilts fed l -carnitine. Free and total carnitine concentration increased (p < 0.05) in the skeletal muscle from the foetuses collected from gilts fed supplemental l -carnitine. This study showed that l -carnitine had beneficial effects on the average foetal weight at day 70 of gestation, associated with changes in the foetal IGF system.     

Stimulatory effect of cold acclimation on skeletal muscle branched-chain α-keto acid dehydrogenase in rats

The effects of cold-acclimation (3 weeks at 4°C) on the actual activities of branched-chain α-keto acid dehydrogenase (BCKDH) and pyruvate dehydrogenase (PDH) complexes in the skeletal muscle and liver of normal 6-week-old rats were studied. The activities of BCKDH and PDH complexes were assayed using [1–¹⁴C] α-ketoisocaproate and [1-¹⁴C]pyruvate, respectively. Cold-acclimation markedly stimulated the activity of BCKDH, and slightly but significantly increased the activities of PDH and citrate synthase (CS) in the skeletal muscle, but did not alter PDH, BCKDH or CS activities in the liver. The increase in muscle BCKDH activity (control rats, 1.45 ± 0.54 mU/g wet wt; cold-acclimated rats, 2.54 ± 0.50 mU/g wet wt; 75% increase: P  = 0.001) was much greater than the increases in PDH activity (84 ± 16 mU/g wet wt and 111 ± 25 mU/g wet wt; 32% increase: P  = 0.020) or CS activities (27 ± 3 μmol/min per g wet wt and 31 ± 2 μmol/min per g wet wt; 14% increase: P  = 0.046). These results suggest that, unlike PDH, BCKDH is not directly associated with the mitochondrial oxidative activity of the skeletal muscle of cold-acclimated rats. This study provides the first evidence that BCKDH in skeletal muscle responds selectively to cold-acclimation.     

Influence of l‐carnitine and l‐arginine on protein synthesis and maturation of the semitendinosus muscle of lightweight piglets

Lightweight (LW ) piglets from large litters display impaired growth performance compared with heavier littermates. This study investigated the growth performance and muscle development of early‐weaned LW piglets (birthweight <1.2 kg) from large litters (17.3 ± 3.0 total born per litter), fed ad libitum a milk replacer supplemented with either l ‐carnitine (CAR ) or l ‐arginine (ARG ) from day 7 to day 28 of age. In total, 36 female and entire male Swiss Large White piglets, weaned on day 7 of age, were artificially reared in pairs in rescue decks. They were allocated to one of three dietary treatments: unsupplemented control (CON ), 0.48 g l ‐carnitine·piglet^?1·day^?1 (CAR ) or 1.20 g l ‐arginine·kg body weight^?1·day^?1 (ARG ). Milk replacer was prepared daily in a 1:4 powder‐to‐water ratio and fed ad libitum. Piglets were weighed at birth and on days 7, 14, 21 and 28. Feed intake was assessed daily. Piglets were euthanized on day 28. The entire semitendinosus muscle (STM ) was collected, and organs were weighed. Subsequently, the STM was divided into the light (STM _l) and dark (STM _d) portion, and contractile and metabolic traits were analysed by ATP histochemistry, enzyme activities and gene expression. No differences in growth performance, organ and STM weight and on contractile traits were found between groups. A tendency (p  <  .10) for an elevated lipid oxidation enzyme activity in the STM _l and STM _d and greater (p  <  .05) phosphorylation of the mammalian target of rapamycin pathway in the STM _l of CAR compared with CON piglets was found. Despite these metabolic responses, the lack of effect of CAR and ARG supplementation on growth performance suggests that providing the milk replacer ad libitum in combination with added CAR and ARG is insufficient for eliciting faster growth of LW piglets.     

Effect of nutrient intake on intramuscular glucose metabolism during the early growth stage in cross‐bred steers (Japanese Black male × Holstein female)

The objective was to investigate the impact of nutrient intake during the early growth period on the expression of glucose metabolism‐related genes in skeletal muscle of cross‐bred cattle. From 1.5 to 5 months of age, group H (n = 7) animals were intensively fed a high‐protein and low‐fat milk replacer [crude protein (CP) 28%; ether extracts (EE) 18%; max: 2.0 kg, 12 l/day], and group R (n = 7) animals were fed a restricted amount of normal milk replacer (CP 25%; EE 23%; max 0.5 kg, 4 l/day). From 6 to 10 months of age, group H cattle were fed a high‐nutrition total mixed ration mainly prepared from grain feed, and group R cattle were fed only roughage. Blood samples were taken from each animal at three biopsy times (1.5, 5 and 10 months of age), and the blood plasma concentration of glucose and insulin was analysed. In glucose concentration, there were no significant differences; however, the concentrations of insulin were higher in group H than in group R at 5 and 10 months of age. Muscle samples were taken by biopsy from longissimus thoracis muscle (LT) at 1.5, 5 and 10 months of age. We analysed mRNA expression levels using the quantitative real‐time polymerase chain reaction (PCR) assay for glucose transporters (GLUT1 and GLUT4), insulin receptor, phosphatidylinositol 3‐kinase (PI‐3K), protein kinase B (PKB, also known as Akt), hexokinase 1 (HK1) and tumour necrosis factor alpha (TNFα). Although no differences were detected at 1.5 and 5 months of age, at 10 months of age, GLUT1, HK1 and TNFα mRNA expression levels were significantly higher in group H than in group R. These results suggested Glut1 that affects insulin‐independently mediated glucose uptake was more responsive to improved nutrition during early growth stage than GLUT4 that insulin‐dependently mediated glucose uptake in LT of cattle.     

Total serum bile acid concentrations in dairy cows with fatty liver and liver failure

In forty-five Holstein Frisian dairy cows (1-6 weeks post partum; mean age: 5.1 +/- 1.2 years) the serum total bile acid concentrations (SBA) were measured enzymatically. In all cows a left sided abomasal displacement was corrected surgically by right side laparotomy and omentopexy three days before investigation. The liver fat content was determined in all cows histologically. Liver failure was assumed if typical clinical signs (ataxia, general depression, recumbency or coma), an increased venous plasma ammonia level (> 35 mumol/l) and a decreased plasma amino acid index (< 4.0) were found. Cows without liver failure (N = 29) were grouped according to the liver fat content as cows with mild (N = 5), moderate (N = 19) or severe hepatosteatosis (N = 5). Histological examination of liver biopsies in cows with liver failure (N = 16) revealed in twelve cases a severe fatty liver and in four cases a hydropic degeneration of the liver tissue. Although in cows without liver failure mean SBA concentrations were higher in the group with moderate (47.3 +/- 30.9 mumol/l) or severe fatty liver (32.9 +/- 21.7 mumol/l) than in that with mild lipidosis (18.0 (16.8 mumol/l), differences were not significant. The mean SBA concentration in cows with liver failure (70.5 +/- 49.5 mumol/l) was only significantly (p < 0.05) increased compared to cows with uncomplicated mild hepatic lipidosis. In conclusion, the determination of SBA concentrations is of little value in the recognition of fatty liver or even liver failure due to the considerable variance of SBA concentrations in dairy cows.     

Coordinated gene expression in adipose tissue and liver differs between cows with high or low NEFA concentrations in early lactation

Dairy cows with high and low plasma non-esterified fatty acid (NEFA) concentrations in early lactation were compared for plasma parameters and mRNA expression of genes in liver and subcutaneous adipose tissue. The study involved 16 multiparous dairy cows with a plasma NEFA concentration of >500 μmol/l [n = 8, high NEFA (HNEFA)] and <140 μmol/l [n = 8, low NEFA (LNEFA)] in the first week post-partum (pp). Blood samples, adipose and liver tissues were collected on day 1 (+1d) and at week 3 pp (+3wk). Blood plasma was assayed for concentrations of metabolites and hormones. Subcutaneous adipose and liver tissues were analysed for mRNA abundance by real-time qRT-PCR encoding parameters related to lipid metabolism. Results showed that mean daily milk yield and milk fat quantity were higher in HNEFA than in LNEFA cows (p < 0.01), and the NEB was more negative in HNEFA than in LNEFA in +3wk too (p < 0.05). HNEFA cows had slightly lower (p < 0.1) insulin concentrations than LNEFA cows across the study period, and the body condition score decreased more from +1d to +3wk in HNEFA than in LNEFA (p = 0.09). The mRNA abundance of genes in the liver related to fatty acid oxidation (carnitine palmitoyltransferase 2 and very long chain acyl-coenzyme A dehydrogenase) and ketogenesis (3-hydroxy-3-methylglutaryl-coenzyme A synthase 2) were lower in HNEFA than in LNEFA cows. No differences between the two groups were observed for mRNA expression of genes in adipose tissue. The number of calculated significant correlation coefficients (moderately strong) between parameters in the liver and in adipose tissue was nearly similar on +1d, and higher for HNEFA compared with LNEFA cows in +3wk. In conclusion, dairy cows with high compared with low plasma NEFA concentrations in early lactation show differentially synchronized mRNA expression of genes in adipose tissue and liver in +3wk that suggests a different orchestrated homeorhetic regulation of lipid metabolism.