Effects of long‐term microalgae supplementation on muscle microstructure,meat quality and fatty acid composition in growing pigs

We investigated the effects of long‐term microalgae supplementation (7% in a piglet diet and 5% in a fattening diet) on muscle microstructure and meat quality, including fatty acid composition in female Landrace pigs (n = 31). The major effects were muscle‐specific increases in n‐3 and n‐6 polyunsaturated fatty acids (PUFA) concentrations, resulting in increased accumulation of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Carcass traits and meat quality of longissimus thoracis muscle were not affected by the microalgae diet with the exception of reduced drip loss (p = 0.01) and increased protein proportion (p = 0.04). In addition, the microalgae diet resulted in a shift to a more oxidative myofibre type composition in semitendinosus but not longissimus thoracis muscle. In conclusion, microalgae supplementation offers a unique opportunity to enhance essential n‐3 PUFA contents in pig meat. The results support small but coordinated changes in skeletal muscle phenotypic appearance and functionality.     

Effects of low and high levels of maternal nutrition consumed for the entirety of gestation on the development of muscle,adipose tissue,bone, and the organs of Wagyu cattle fetuses

This study aimed to investigate the effects of high and low levels of energy intake during the entire gestation period on the skeletal muscle development, organ development, and adipose tissue accumulation in fetuses of Wagyu (Japanese Black) cows, a breed with highly marbled beef. Cows were allocated to a high-nutrition (n = 6) group (fed 120% of the nutritional requirement) or low-nutrition (n = 6) group (fed 60% of the nutritional requirement). The cows were artificially inseminated with semen from the same sire, and the fetuses were removed by cesarean section at 260 ± 8.3 days of fetal age and slaughtered. The whole-body, total muscle, adipose, and bone masses of the fetal half-carcasses were significantly higher in the high-nutrition group than the low-nutrition group (p = 0.0018, 0.009, 0.0004, and 0.0362, respectively). Fifteen of 20 individual muscles, five of six fat depots, nine of 17 organs, and seven of 12 bones that were investigated had significantly higher masses in the high-nutrition group than the low-nutrition group. The crude components and amino acid composition of the longissimus muscle significantly differed between the low- and high-nutrition groups. These data indicate that maternal nutrition during gestation has a marked effect on the muscle, bone, and adipose tissue development of Wagyu cattle fetuses.     

Muscle type‐specific effect of myostatin deficiency on myogenic regulatory factor expression in adult double‐muscled Japanese Shorthorn cattle

To clarify muscle type‐specific effect of myostatin on myogenic regulatory factors (MRFs), we examined mRNA expression of MRFs in five skeletal muscles of normal (NM) and myostatin‐deficient double‐muscled (DM) adult Japanese Shorthorn cattle by quantitative reverse‐transcribed PCR. Among the four MRFs, namely, Myf5, MyoD, myogenin, and MRF4, MyoD expression was different among the muscles of the DM cattle (P < 0.01) but not of the NM cattle. Meanwhile, MyoD expression was significantly elevated only in masseter (MS) muscle in the DM cattle due to the myostatin deficiency (P < 0.05). Myf5 and MRF4 expression in semitendinosus (ST) was higher in the DM than in the NM cattle (P < 0.05). According to analysis of myosin heavy chain (MyHC) isoform expression, more MyHC‐2x and ‐2a and less ‐slow isoforms were expressed in the longissimus and ST muscles compared to the MS muscle in both cattle (P < 0.05), but no significant difference in MyHC expression was observed between the NM and DM cattle. Taken together, myostatin has influences on Myf5 and MRF4 expression in faster‐type muscles and on MyoD expression in slower‐type muscles, suggesting a possible muscle type‐specific effect of myostatin in skeletal muscle growth and maintenance.     

Relationships between thyroid status, tissue oxidative metabolism, and muscle differentiation in bovine fetuses

The temporal relationships between thyroid status and differentiation of liver, heart and different skeletal muscles were examined in 42 bovine fetuses from day 110 to day 260 of development using principal component analysis of the data. Plasma concentrations of reverse-triiodothyronine (rT(3)) and thyroxine (T(4)) increased during development from day 110 to day 210 or 260, respectively, whereas concentration of triiodothyronine (T(3)) and hepatic type-1 5'-deiodinase activity (5'D1) increased from day 180 onwards. On day 260, high T(4) and rT(3) and low T(3) concentrations were observed together with a mature 5'D1 activity. Cytochrome-c oxidase (COX) activity expressed per mg protein increased at day 180 in masseter and near birth in masseter, rectus abdominis and cutaneus trunci muscles (P<0.05). Significant changes in citrate synthase (CS) activity per mg protein were observed between day 110 and day 180 in the liver and between day 210 and day 260 in the liver, the heart and the longissimus thoracis muscle (P<0.05). Muscle contractile differentiation was shown by the disappearance of the fetal myosin heavy chain from day 180 onwards. A positive correlation (r>0.47, P<0.01) was shown between thyroid status parameters (5'D1, concentrations of T(4) and T(3)) and COX activity in muscles known to be oxidative after birth (masseter, rectus abdominis) but not in liver and heart, nor in muscles known to be glycolytic after birth (cutaneus trunci, longissimus thoracis). A similar correlation was found between thyroid parameters and CS activity in liver and masseter. Results indicate that elevation of plasma T(3) concentrations in the last gestational trimester could be involved in the differentiation of oxidative skeletal muscles.     

Identification of polymorphisms and association analysis with meat quality traits in the porcine KIAA1717 and HUMMLC2B genes

Skeletal muscle genes are potential candidates for production and meat quality. Screening a subtracted cDNA library constructed with mRNA obtained from longissimus dorsi muscles of F1 hybrids Landrace × Yorkshire and their female parents Yorkshire, we isolated two partial sequences coding for the H3-K4-specific methyltransferase (KIAA1717) and skeletal muscle myosin regulatory light chain (HUMMLC2B) genes. Database search revealed KIAA1717 and HUMMLC2B encoded proteins with SET domain and EF-hand calcium binding motif, respectively. In the present work we identified their partial polymorphisms and two SNPs, one (C1354T) at the 3′ untranslated region (UTR) of KIAA1717 and one (A345G) at the SINE (PRE-1) element of HUMMLC2B, both created/disrupted a restriction site for endonuclease Msp I. The selected pigs were genotyped at the KIAA1717 C1354T and HUMMLC2B A345G sites by means of a PCR-RFLP protocol. Significant associations were observed for the KIAA1717 C1354T polymorphic site with meat marbling (longissimus doris (p < 0.05), biceps femoris (p < 0.01)) and intramuscular fat (p < 0.01). HUMMLC2B A345G were significantly associated with meat pH (longissimus doris (p < 0.05), biceps femoris (p < 0.01)), drip loss (p < 0.01), water holding capacity (p < 0.01) and meat color value (longissimus doris (p < 0.01), biceps femoris (p < 0.05)). Further studies are needed to confirm these preliminary results.     

The influence of the PRKAG3 mutation on glycogen,enzyme activities and fibre types in different skeletal muscles of exercise trained pigs

Background

AMP-activated protein kinase (AMPK) plays an important role in the regulation of glucose and lipid metabolism in skeletal muscle. Many pigs of Hampshire origin have a naturally occurring dominant mutation in the AMPK γ3 subunit. Pigs carrying this PRKAG3 (R225Q) mutation have, compared to non-carriers, higher muscle glycogen levels and increased oxidative capacity in m. longissimus dorsi, containing mainly type II glycolytic fibres. These metabolic changes resemble those seen when muscles adapt to an increased physical activity level. The aim was to stimulate AMPK by exercise training and study the influence of the PRKAG3 mutation on metabolic and fibre characteristics not only in m. longissimus dorsi, but also in other muscles with different functions.

Methods

Eight pigs, with the PRKAG3 mutation, and eight pigs without the mutation were exercise trained on a treadmill. One week after the training period muscle samples were obtained after euthanisation from m. biceps femoris, m. longissimus dorsi, m. masseter and m. semitendinosus. Glycogen content was analysed in all these muscles. Enzyme activities were analysed on m. biceps femoris, m. longissimus dorsi, and m. semitendinosus to evaluate the capacity for phosphorylation of glucose and the oxidative and glycolytic capacity. Fibre types were identified with the myosin ATPase method and in m. biceps femoris and m. longissimus dorsi, immunohistochemical methods were also used.

Results

The carriers of the PRKAG3 mutation had compared to the non-carriers higher muscle glycogen content, increased capacity for phosphorylation of glucose, increased oxidative and decreased glycolytic capacity in m. longissimus dorsi and increased phosphorylase activity in m. biceps femoris and m. longissimus dorsi. No differences between genotypes were seen when fibre type composition was evaluated with the myosin ATPase method. Immunohistochemical methods showed that the carriers compared to the non-carriers had a higher percentage of type II fibres stained with the antibody identifying type IIA and IIX fibres in m. longissimus dorsi and a lower percentage of type IIB fibres in both m. biceps femoris and m. longissimus dorsi. In these muscles the relative area of type IIB fibres was lower in carriers than in non-carriers.

Conclusions

In exercise-trained pigs, the PRKAG3 mutation influences muscle characteristics and promotes an oxidative phenotype to a varying degree among muscles with different functions.     

Duration of feeding linseed diet influences peroxisome proliferator-activated receptor γ and tumor necrosis factor gene expression, and muscle mass of growing–finishing barrows

The aim of the study was to investigate the effect of duration of feeding linseed (rich in n-3 PUFA) on peroxisome proliferator-activated receptor γ (PPARγ) and tumor necrosis factor (TNF-α) gene expression, and muscle mass of growing–finishing barrows. Two isoenergetic and isonitrogenous diets were formulated, and one of which was the basal diet and another one was the linseed diet including linseed at the level of 10%. Twenty-four Landrace × Yorkshire barrows weighing 35 ± 3.7 kg were randomly assigned to four treatments with six individuals per treatment. Pigs in treatment 1 (T1) fed the control diet throughout the experimental period, while pigs in T2, T3 and T4 fed the control diet except for 30, 60, and 90 d prior to slaughter when the linseed diet were fed. The experiment was conducted for 90 days. The longissimus muscle mass and each muscle mass in the hind leg were weighted. PPARγ and TNF-α mRNA expression levels in muscle, spleen and adipose tissue, and plasma concentrations of TNF-α data were measured and analyzed. The results showed that the longissimus muscle mass, quadriceps femoris muscle mass and semitendinosus muscle mass increased linearly (P < 0.01) as prolonged the time of feeding linseed diet. The expression of PPARγ in longissimus muscle and spleen increased (P < 0.01) linearly as prolonged the time of feeding linseed diet, while the expression of PPARγ in adipose tissue were not affected (P = 0.095). Duration of linseed addition linearly decreased (P < 0.01) TNF-α gene expression levels in the longissimus dorsi muscle, adipose and spleen, and serum concentration of TNF-α as well. The expression levels of PPARγ negatively correlated with the expression of TNF-α in muscle (R² = 0.70, P < 0.001) and spleen (R²R² = 0.77, P < 0.001) respectively. Likewise, PPARγ expression level in spleen (R²R² = 0.59, P < 0.01) or muscle (R²R² = 0.52, P < 0.05) negative correlated with serum TNF-α concentration, while there were significant quadratic relation between muscular PPARγ (R²R² = 0.80, P < 0.01) or muscular TNF-α (R²R² = 0.87, P < 0.01) expression and the longissimus dorsi muscle mass. These data suggested that duration of feeding linseed diet lead to a linear decrease of TNF-α gene expression, which may increase the muscle mass in growing–finishing barrows, at least in part, through a PPARγ-dependent mechanism.     

Occurrence of Blood Cells and Serum Proteins in Bovine Fetuses and Calves

Hematological values of peripheral blood were determined for bovine fetuses and calves of various ages. Erythrocyte values increased through gestation. Fetuses 100 days or older had total values within the ranges of those reported for normal adult cattle. Mature erythrocytes were not observed in embryos and only a few were observed in fetuses 40 days of age. Fetuses 250 days or older had only a few rubricytes (<10/100 WBC). Leukocytes were first identified in the peripheral blood of a 45-day old fetus. Absolute leukocyte values increased through gestation and reached maximum values shortly before parturition. Granulocytes were first observed at 130 days of gestation and reached maximum values near parturition.

Total serum protein and gamma-globulin concentrations of colostrum-deprived calves were similar to serum protein and gammaglobulin concentrations of fetuses older than 265 days and were lower than values for the colostrum-fed calves. The immunoelectrophoretic pattern of 59-day old fetuses, the earliest age at which serum samples were obtained, demonstrated albumin, an α1 globulin and a β globulin, possibly transferrin. Additional α and β globulins appeared in the older fetuses and by 175 days of gestation serum electophoretic patterns of the fetuses were similar to patterns normally found with adult bovine serum except for the absence of the gammaglobulins in fetal serum. Immunoglobulin M was detected in 39 of 95 fetal serum samples by radial diffusion and in 13 of 95 samples by immunoelectrophoresis. Immunoglobulin G was detected in ten of 95 fetal serum samples by radial diffusion and in six of 95 samples by immunoelectrophoresis.

     

Canine thyrotropin β-subunit gene: cloning and expression in Escherichia coli, generation of monoclonal antibodies, and transient expression in the Chinese hamster ovary cells

The gene encoding the mature β subunit of canine thyroid stimulating hormone (cTSHβ) was cloned, sequenced and expressed in Escherichia coli and in Chinese hamster ovary (CHO) cells, and monoclonal antibodies against the recombinant cTSHβ purified from E. coli were generated. The gene fragment that encodes mature TSHβ was cloned from the canine genomic DNA by direct polymerase chain reaction (PCR) using primers that were designed based on the consensus sequences from other species. The resulting 891 basepairs (bp) of genomic DNA consisted of two coding exons of the canine TSHβ gene and an intron of 450 bp. The two exons, which encode the mature cTSHβ subunit, was joined together by an overlap PCR and was expressed in E. coli as 6×His-tagged protein. The purified recombinant cTSHβ with a molecular weight of about 15 kDa was recognized by the polyclonal antibodies prepared against the native canine TSH in Western blot. Monoclonal antibodies were raised against the purified cTSHβ and subsequently characterized. For transient expression in CHO cells that are permanently transfected with the bovine common α gene, a 60-oligonucleotide signal peptide coding sequence was added to the 5′ end of the cTSHβ gene before it was cloned into the mammalian expression vector pRSV and used to transfect CHO cells. The medium from these transfected cells, presumably containing the bovine α and canine TSHβ in heterodimeric confirmation, exhibited TSH bioactivity as indicated by the stimulation of cAMP production in the cultured FRTL-5 thyrocytes.     

Levels of mRNA for three selenoproteins in skeletal muscle of fetal and newborn pigs

Selenoproteins have wide-ranging functions throughout the body, including important roles in protection against oxidative stress, as well as muscle development and integrity. In this study, prepubertal gilts were randomly assigned to either a Se-sufficient or deficient diet (n = 12/treatment). Levels of mRNA coding for the selenoproteins glutathione peroxidase-1 (GPx-1), selenoprotein W (SelW) and selenoprotein N (SelN) and the concentration of Se in fetal and neonatal swine skeletal muscle (gluteus maximus) were measured at time intervals throughout gestation and immediately after birth. Total RNA was isolated from skeletal muscle, and cDNA was synthesized for real-time RT-PCR analysis of the selected selenoproteins. Selenium concentration in skeletal muscle of fetal and newborn pigs decreased throughout gestation (P < 0.0001) and was lower (P < 0.05) when dams were fed the low Se diet. Levels of GPx-1 mRNA in fetal skeletal muscle were unaffected by maternal Se intake nor did GPx-1 mRNA levels change during late gestation. Selenoprotein W mRNA levels increased (P = 0.01) in fetal skeletal muscle during late gestation but were reduced (P = 0.05) by low maternal intake of Se. Levels of SelN mRNA showed a gestational time by treatment interaction (P < 0.05). The increased levels of SelW mRNA in skeletal muscle during late gestation, despite a decrease in Se concentration, indicate that SelW may be an important antioxidant protein to the late term fetus and the newborn.     

Comparison of enzyme activities and gene expression profiling between yak and bovine skeletal muscles

The objective of the present study was to reveal the differences between yak (Bos grunniens) and cattle in energy metabolic characteristics and gene expressions in skeletal muscles. Activities of lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and β-hydroxyacyl-CoA dehydrogenase (HOAD), which are involved in metabolism of carbohydrate and fatty acid respectively, were measured in longissimus dorsi and biceps femoris. Yak contained higher total LDH activity and higher proportion of LDH5 in longissimus dorsi than cattle (P < 0.05), indicating its more anaerobic potential characteristics in carbohydrate metabolism. However, yak contained higher activities of MDH and HOAD (P < 0.05) in biceps femoris than cattle, exhibiting high oxidative capacity under hypoxic environment. Semi-quantitative RT-PCR analysis of mRNA abundance showed that yak and cattle contained a similar level of adipocyte fatty acid-binding protein (A-FABP) in both longissimus dorsi and biceps femoris, while myosin heavy chain I (MyHC I) in longissimus dorsi and biceps femoris, and peroxisome proliferator-activated receptor coactivator 1α (PGC-1α) level in biceps femoris were significant lower in yak compared with cattle (P < 0.01 and P < 0.05 respectively), indicating the anaerobic characteristic of yak skeletal muscles. In conclusion, our experiment showed that yak skeletal muscles exhibit different energy metabolic property and gene expressions compared to cattle.     

Niacin supplementation induces type II to type I muscle fiber transition in skeletal muscle of sheep

Background

It was recently shown that niacin supplementation counteracts the obesity-induced muscle fiber transition from oxidative type I to glycolytic type II and increases the number of type I fibers in skeletal muscle of obese Zucker rats. These effects were likely mediated by the induction of key regulators of fiber transition, PPARδ (encoded by PPARD), PGC-1α (encoded by PPARGC1A) and PGC-1β (encoded by PPARGC1B), leading to type II to type I fiber transition and upregulation of genes involved in oxidative metabolism. The aim of the present study was to investigate whether niacin administration also influences fiber distribution and the metabolic phenotype of different muscles [M. longissimus dorsi (LD), M. semimembranosus (SM), M. semitendinosus (ST)] in sheep as a model for ruminants. For this purpose, 16 male, 11 wk old Rhoen sheep were randomly allocated to two groups of 8 sheep each administered either no (control group) or 1 g niacin per day (niacin group) for 4 wk.

Results

After 4 wk, the percentage number of type I fibers in LD, SM and ST muscles was greater in the niacin group, whereas the percentage number of type II fibers was less in niacin group than in the control group (P < 0.05). The mRNA levels of PPARGC1A, PPARGC1B, and PPARD and the relative mRNA levels of genes involved in mitochondrial fatty acid uptake (CPT1B, SLC25A20), tricarboxylic acid cycle (SDHA), mitochondrial respiratory chain (COX5A, COX6A1), and angiogenesis (VEGFA) in LD, SM and ST muscles were greater (P < 0.05) or tended to be greater (P < 0.15) in the niacin group than in the control group.

Conclusions

The study shows that niacin supplementation induces muscle fiber transition from type II to type I, and thereby an oxidative metabolic phenotype of skeletal muscle in sheep as a model for ruminants. The enhanced capacity of skeletal muscle to utilize fatty acids in ruminants might be particularly useful during metabolic states in which fatty acids are excessively mobilized from adipose tissue, such as during the early lactating period in high producing cows.     

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.     

Inflammatory infiltration into placentas of Neospora caninum challenged cattle correlates with clinical outcome of pregnancy

Infection with Neospora caninum stimulates host cell-mediated immune responses, which may be responsible for placental damage leading to bovine abortion. The aim of this study was to compare immune responses in the bovine placenta, following experimental infection in different stages of pregnancy. Placentomes were examined by immunohistochemistry and inflammation in early gestation was generally moderate to severe, particularly in the placentas carrying non-viable foetuses, whereas it was milder in later stages, mainly characterised by the presence of CD3⁺, CD4⁺ and γδ T-cells. This distinctive cellular immune response may explain the milder clinical outcome observed when animals are infected in later gestation.     

Effects of Dietary Supplementation of β‐hydroxy‐β‐methylbutyrate on Sow Performance and mRNA Expression of Myogenic Markers in Skeletal Muscle of Neonatal Piglets

The effects of dietary β‐hydroxy‐β‐methylbutyrate (HMB) supplementation during gestation on reproductive performance of sows and the mRNA expression of myogenic markers in skeletal muscle of neonatal pigs were determined. At day 35 of gestation, a total of 20 sows (Landrace × Yorkshire, at third parity) were randomly assigned to two groups, with each group receiving either a basal diet or the same diet supplemented with 4 g/day β‐hydroxy‐β‐methylbutyrate calcium (HMB‐Ca) until parturition. At parturition, the total and live litter size were not markedly different between treatments, however, the sows fed HMB diet had a decreased rate of stillborn piglets compared with the sows fed the control (CON) diets (p < 0.05). In addition, piglets from the sows fed HMB diet tended to have an increased birth weight (p = 0.08), and a reduced rate of low birth weight piglets (p = 0.05) compared with piglets from the CON sows. Nevertheless, lower feed intake during lactation was observed in the sows fed the HMB diet compared with those on the CON diet (p < 0.01). The relative weights of the longissimus dorsi (LD) and semitendinosus (ST) muscle were higher (p < 0.05) in neonatal pigs from the HMB than the CON sows. Furthermore, maternal HMB treatment increased the mRNA levels of the myogenic genes, including muscle regulatory factor‐4 (MRF4, p < 0.05), myogenic differentiation factor (MyoD) and insulin‐like growth factor‐1 (IGF‐1, p < 0.01). In conclusion, dietary HMB supplementation to sows at 4 g/day from day 35 of gestation to term significantly improves pregnancy outcomes and increases the expression of myogenic genes in skeletal muscle of neonatal piglets, but reduces feed intake of sows during lactation.     

The expression pattern,polymorphisms and association analyses of the porcine NREP gene

NREP (neuronal regeneration related protein homolog) plays a role in the transformation of neural, muscle, and fibroblast cells and in smooth muscle myogenesis. The NREP gene was selected for detailed study as an expressional and functional candidate gene on the basis of data from the expression microarray, which detected the differences in gene expression between Czech Large White pigs and wild boars in the longissimus lumborum et thoracis and biceps femoris muscle tissues. Quantitative real-time PCR results confirmed that porcine NREP was expressed in both skeletal muscles and significantly overexpressed in Czech Large White pigs compared with wild boars (14.5- and 11.6-fold; p < .05). We identified 9 polymorphic sites in the genomic DNA of NREP. Six of these polymorphisms were in complete linkage disequilibrium, and therefore, only 4 loci were informative. The associations of the HF571253:g.103G>A, HF571253:g.134G>A, HF571253:g.179T>C and HF571253:g.402_409delT polymorphisms with backfat thickness, lean meat content and average daily gain were assessed in Czech Large White pigs. The GG genotypes HF571253:g.103G>A and HF571253:g.134G>A, the TT genotypes HF571253:g.179T>C and 67 HF571253:g.402_409delT genotypes had favourable effects on the studied traits. Our results indicate the possibility of utilizing the variability of the NREP gene in marker-assisted selection in order to improve meat production in pigs.     

Interrelationship of growth hormone AluI polymorphism and hyperketonemia with plasma hormones and metabolites in the beginning of lactation in dairy cows

A polymorphic site of the growth hormone gene (AluI polymorphism) that results in an amino acid change at position 127 of the protein chain (leucine, L to valine, V) has been linked to differences in circulating metabolites and metabolic hormones of calves and bulls and to milk yield traits of lactating cows. Our objective was to investigate the interrelationship of this polymorphism with plasma concentrations of β-hydroxybutyrate, insulin, IGF-I and leptin in postpartum dairy cows. Blood samples were taken from clinically healthy, spring-calving, group-fed Holstein-Friesian cows (n = 257; 7 large-scale farms) 4–13 days after calving. Of all herds, 100 cows had plasma β-hydroxybutyrate levels above 1.2 mmol/l and 157 cows were normoketonemic. The proportion of valine carriers and LL cows was not different within groups of normo- and hyperketonemic animals. Genotype was not associated with plasma β-hydroxybutyrate, insulin, IGF-I and leptin levels either in all of the herds or in the two with the highest proportion of the valine allele carriers (n = 28, 72%). We found significantly lower insulin, IGF-I and leptin concentrations in the presence of hyperketonemia compared to normoketonemic cows. There were strong negative correlations between BHB and the other blood parameters, while insulin, IGF-I and leptin were positively related to each other. In conclusion, in the first two weeks after calving we could not demonstrate any effect of AluI polymorphism on plasma concentrations of β-hydroxybutyrate and metabolic hormones studied. Hyperketonemia was associated with a significant decrease in insulin, IGF-I and leptin blood levels. We infer that cows homozygous for the leucine allele or carrying the valine allele may have a similar endocrine and metabolic response to the challenge of increased nutrient demand early postpartum and that the presence of hyperketonemia is mainly linked to the hormonal and metabolic changes occurring at the onset of lactation.     

Insulin resistance: Relationship between indices during late gestation in dairy cows and effects on newborn metabolism

To investigate the relationship between insulin resistance indices [“Revised quantitative insulin sensitivity check index” (RQUICKI; RQ), “Revised quantitative insulin sensitivity check index ‐ β‐hydroxybutyrate” (RQUICKI_BHB; RQ_BHB), and “Homeostasis model assessment‐insulin resistance” (HOMA‐IR; HR)], and metabolic parameters in dams during late gestation, and their newborn calves. Blood was sampled twice weekly during the experimental period in 30 dry Holstein cows. In calves, blood sampling and body weight measurements were performed immediately after birth, and in 1‐week‐old male calves, liver and muscle biopsy samples were obtained for determining metabolic factor mRNA levels. RQ and RQ_BHB were negatively correlated with insulin, nonesterified fatty acid, BHB, and albumin and were positively correlated with leptin levels in blood during late gestation (p < .05). RQ, rather than RQ_BHB, reflected metabolism of dams, while stronger positive correlations were present between HR and blood insulin concentrations than other parameters, and calves of dams with high HR had low body weight, and high liver and muscle expression of growth hormone and insulin receptor mRNA (p < .05). RQ and HR of dams during late gestation could serve as indicators of dam metabolism and predictors of metabolism in newborn calves respectively.     

Phenotypic characterisation of the cellular immune infiltrate in placentas of cattle following experimental inoculation with Neospora caninum in late gestation

Despite Neospora caninum being a major cause of bovine abortion worldwide, its pathogenesis is not completely understood. Neospora infection stimulates host cell-mediated immune responses, which may be responsible for the placental damage leading to abortion. The aim of the current study was to characterize the placental immune response following an experimental inoculation of pregnant cattle with N. caninum tachyzoites at day 210 of gestation. Cows were culled at 14, 28, 42 and 56 days post inoculation (dpi). Placentomes were examined by immunohistochemistry using antibodies against macrophages, T-cell subsets (CD4, CD8 and γδ), NK cells and B cells. Macrophages were detected mainly at 14 days post inoculation. Inflammation was generally mild and mainly characterized by CD3⁺, CD4⁺ and γδ T-cells; whereas CD8⁺ and NK cells were less numerous. The immune cell repertoire observed in this study was similar to those seen in pregnant cattle challenged with N. caninum at early gestation. However, cellular infiltrates were less severe than those seen during first trimester Neospora infections. This may explain the milder clinical outcome observed when animals are infected late in gestation.     

The influence of different doses of α-tocopherol and ascorbic acid on selected oxidative stress parameters in in vitro culture of leukocytes isolated from transported calves

The objective of the study was to assess in vitro the influence of various doses of two different antioxidants, α-tocopherol and ascorbic acid, on protective mechanisms against ROS in white blood cells obtained from calves exposed to transport, and to compare these results with those obtained from non-transported animals.The concentrations of lipid peroxidation products in leukocytes and in the retained medium were assessed by determining the level of ThioBarbituric Acid Reactive Substances (TBARS). Total antioxidant status in the leukocytes and the medium were estimated using a ferric-reducing ability of plasma (FRAP) assay. Leukocyte viability was determined using the trypan blue reduction test.The study demonstrated that after bovine leukocytes (WBC) were incubated in vitro with α-tocopherol and ascorbic acid, peroxidation intensity decreased and total antioxidant capacity increased. The results of the study reveal that these antioxidants in concentrations over 0.1 mg/ml have a major impact on free radical activity on bovine white blood cells and on cell viability during transport of animals.Based on this study, we suggest that incubation of the leukocytes with antioxidants decreases the oxidative stress development, which can be helpful in protection of the immunological cells during bovine respiratory disease.