Investigation of single‐nucleotide polymorphisms in the NR3C1a glucocorticoid receptor gene in Cocker Spaniels with primary immune thrombocytopenia

BackgroundIn dogs, 6 single‐nucleotide polymorphisms (SNPs) have been described in the glucocorticoid receptor gene NR3C1a, 2 of which were nonsynonymous SNPs in exons 2 and 8. The clinical importance of these SNPs is unknown.ObjectivesTo investigate whether SNPs in NR3C1a are associated with clinical outcome in Cocker Spaniels with primary immune thrombocytopenia (pITP).AnimalsTwenty‐four Cocker Spaniels with pITP presented to a referral center. Dogs were classified as slow (n = 11) or fast responders (n = 12) based on time required after initiating glucocorticoid treatment to achieve a platelet count >70 000/μL.MethodsDeoxyribonucleic acid was extracted from stored blood samples before amplification by PCR and sequencing of exons 2 and 8 of NR3C1a. Associations between genotype and clinical response variables were investigated.ResultsNeither previously identified nonsynonymous SNPs were identified. The synonymous SNP NR3C1a:c.798C>T in exon 2 was found at an increased prevalence compared to a previous report. No difference was found in prevalence of any genotype at NR3C1a:c.798C>T between fast and slow responders (P = .70).Conclusions and Clinical ImportanceNone of the previously reported nonsynonymous SNPs in exons 2 and 8 of the NR3C1a gene were detected in our cohort of Cocker Spaniels with pITP. The synonymous SNP NR3C1a:c.798C>T in exon 2 was reported at a higher frequency than previously, but was not associated with outcome measures that estimated responsiveness to glucocorticoids.     

Serum α‐1‐acid glycoprotein concentration in clinically healthy puppies and adult dogs and in dogs with various diseases

Background: α‐1‐acid glycoprotein (AGP) is an acute‐phase protein and a serum marker of inflammation and neoplasia in humans. AGP concentrations in diseased dogs and the potential effects of age, breed, and sex have not been elucidated. Objective: The purpose of this study was to examine differences in AGP concentration based on age, sex, and breed in a large population of clinically healthy dogs and to compare AGP concentrations in dogs with various diseases. Methods: Serum was obtained from clinically healthy puppies (n=74) and adults (n=172) of both sexes, and included mongrels (n=205) and Beagles (n=41). Serum also was obtained from 192 dogs with various diseases, including 8 with pyometra that were sampled before, and 1, 2, 3, and 10 days after surgery. AGP concentration was measured by single radial immunodiffusion. Statistical comparisons were made among age, sex, breed, and disease groups. Results: Serum AGP in healthy adult mongrels was 364±106 mg/L (reference interval, 152–576 mg/L). AGP was lowest in newborns (n=11, 122±54 mg/L) and gradually increased to adult levels by 3 months of age. Median AGP concentration was highest in dogs with parvovirus (n=17, 2100 mg/L), distemper (n=7, 1250 mg/L), and pyometra (n=18, 2480 mg/L) and was also significantly higher in dogs with acute filariasis, renal failure, urolithiasis, pancreatitis, hepatitis, trauma, hyperadrenocorticism, and immune‐mediated hemolytic anemia. Dogs with acute filariasis and acute hepatopathy had significantly higher AGP concentrations than dogs with chronic filariasis and chronic hepatopathy. Serum AGP concentration decreased gradually following surgery for pyometra but remained increased after 10 days (896±175 mg/L). Conclusions: Because of significantly lower AGP in puppies, the age of dogs should be considered when using AGP as a marker of disease. Serum AGP may be a useful marker of inflammatory disease in dogs and may help differentiate acute and chronic stages of disease.     

Sexual polymorphisms of vomeronasal 1 receptor family gene expression in bulls,steers, and estrous and early luteal-phase heifers

Vomeronasal 1 receptors (V1R) are a family of receptors for intraspecies chemosignals, including pheromones,and are expressed in the olfactory epithelium (OE) and vomeronasal organ (VO). Even in the well-studiedrodents, it is unclear which members of the V1R family cause sexual polymorphisms, as there are numerous genesand it is difficult to quantify their expressions individually. Bovine species carry only 34 V1R homologs, andthe OE and VOs are large enough to sample. Here, V1R expression was quantified in the OE and VOs of individualbovines. Based on the 34 gene sequences, we obtained a molecular dendrogram consisting of four clusters andsix independent branches. Semi-quantitative RT-PCR was used to obtain gene expression profiles in the VOs andOE of 5 Japanese Black bulls, 5 steers, 7 estrous heifers and 6 early luteal-phase heifers. Ten genes showedsignificant between-group differences, and 22 showed high expression in VOs than in OE. The bulls showedhigher expression of one gene more in OE and another in VOs (both P<0.05) than did steers;both genes belonged to the first cluster. No genes were expressed more abundantly in steers than in bulls. Theestrous heifers showed higher expression of a gene of the second cluster in OE, and a gene of the thirdcluster in VOs (both P<0.05) than did early luteal-phase heifers. These results suggestV1R expression exhibits sexual polymorphisms in cattle.     

The influence of sex and gonadectomy on hepatic and brain fatty acid composition,lipogenesis and β‐oxidation

The aim of this study was to investigate the influence of sex and castration of rats on liver and brain fatty acid profile and liver mRNA expression of genes involved in lipogenesis and β‐oxidation. Castration significantly increased body weight and liver index and decreased serum triglyceride content in the female rats. The fatty acid composition of the liver tissue was influenced by sex and castration. Male rats had higher content of C16:0, C18:1n7, C18:2n6 and C22:5n3, while female rats had higher content of C18:0, C20:4n6 and C22:6n3. Castration of male rats decreased differences caused by sex for C18:2n6, C20:4n6 and C22:6n3. Values for C16:1n7 were higher in the castrated male rats in comparison with all other groups. Liver phospholipids showed a distribution of fatty acids similar to the total lipids. Brain total lipids and phospholipids were not influenced by sex or castration. Castration increased ?6D gene expression in both the sexes, while ?5D and ?9D increased in females and males respectively. Gonadectomy increased expression of the FASN gene in the females and decreased CPT1 and ACOX1 gene expression in the liver tissue of male rats. The observed results of lipid peroxidation, measured by TBARS, were the lowest in the intact females in comparison with all other groups. In conclusion, sex strongly influences both SFA and PUFA in liver tissue, and castration decreases these differences only for PUFA. Castration also influences the expression of the genes involved in lipid metabolism differently in male and female rats, with an increase in lipogenic genes in female rats and a decrease in key genes for mitochondrial and peroxisomal β‐oxidation in male rats.     

Epistatic effects between pairs of the growth hormone secretagogue receptor 1a,growth hormone,growth hormone receptor,non‐SMC condensin I complex,subunit G and stearoyl‐CoA desaturase genes on carcass,price‐related and fatty acid composition traits in Japanese Black cattle

Growth hormone secretagogue receptor 1a (GHSR1a), growth hormone (GH), growth hormone receptor (GHR), non‐SMC condensin I complex, subunit G (NCAPG) and stearoyl‐CoA desaturase (SCD), are known to play important roles in growth and lipid metabolisms. Single and epistatic effects of the five genes on carcass, price‐related and fatty acid (FA) composition traits were analyzed in a commercial Japanese Black cattle population of Ibaraki Prefecture. A total of 650 steers and 116 heifers for carcass and price‐related traits, and 158 steers for FA composition traits were used in this study. Epistatic effects between pairs of the five genes were found in several traits. Alleles showing strain‐specific differences in the five genes had significant single and epistatic effects in some traits. The data suggest that a TG‐repeat polymorphism of the GHSR1a.5′UTR‐(TG)_n locus plays a central role in gene–gene epistatic interaction of FA composition traits in the adipose tissue of Japanese Black cattle.     

Identification of single-nucleotide polymorphisms in 5' end and exons of the PRKAG3 gene in Hubbard White broiler, Leghorn layer, and three Chinese indigenous chicken breeds

5′‐AMP‐activated protein kinase plays an important role in regulating the level of ATP in the presence of metabolic stress. Previous studies revealed that polymorphisms in 5′‐AMP‐activated protein kinase gamma 3 subunit (PRKAG3) gene are associated with meat quality in pigs. In the present study, single‐nucleotide polymorphisms (SNPs) in the 5′‐end and exons of chicken PRKAG3 gene were identified with the method of single‐strand conformation polymorphism in Hubbard ISA White broiler, Leghorn layer, and three Chinese indigenous chicken breeds, Tibet Chicken, Shouguang Chicken and Beijing Yellow Chicken. Two SNPs in the 5′‐end of the gene and 10 SNPs in exons 3, 4, 9 and 11, of which three caused amino acid substitutions, were identified in the PRKAG3 gene of the five chicken breeds. The results will facilitate further study on the association between the mutations of PRKAG3 and chicken meat quality.     

Sidr honey abrogates the oxidative stress and downregulates the hyaluronic acid concentration and gene expression of TGF‐β1 and COL1a1 in rat model of thioacetamide‐induced hepatic fibrosis

Liver fibrosis is a major health concern, which might progress to cirrhosis. To date, treatment trials rely mainly on the removal of the causative factor. The current study investigated the potential ameliorative role of sidr honey on thioacetamide (TAA)‐induced liver fibrosis in rats. Forty‐eight Wistar albino rats were equally allocated into four groups: control; sidr honey (5g/kg body weight (BW), orally); TAA (200 mg/kg BW, IP three times weekly/15 weeks); and sidr honey plus TAA at the same dose and administration rout. Rats co‐treated with sidr honey plus TAA revealed significant reduction in hepatic malondialdehyde, hyaluronic acid (HA), alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, gamma glutamyl transferase, direct bilirubin, and hepatic mRNA expression of transforming growth factor (TGF)‐β1 and collagen type I alpha 1 chain (COL1a1) compared to TAA‐exposed rats. In addition, the hepatoprotective potential of sidr honey was indicated via improvement of histopathologic picture of hepatocytes and upregulation of total antioxidant capacity, reduced glutathione, catalase, glutathione peroxidase, superoxide dismutase, total protein, and albumin compared to TAA‐treated rats. In conclusion, daily administration of sidr honey (5 g/kg BW) is a promising natural antioxidant and fibrosuppressive agent that could ameliorate liver fibrosis via downregulation of fibrosis genes including TGF‐β1 and COL1a1 and HA and via enhancement of antioxidant system.     

Intron 1 of the interferon γ gene: Its role in nematode resistance in Suffolk and Texel sheep breeds

Genetic variation at intron 1 of the interferon gamma gene has recently been associated with variation in faecal egg count in Romney, Merino and Soay breeds of sheep. The Texel breed is more resistant to gastrointestinal nematode infection than the Suffolk breed, based on faecal egg count. Hence, the objective of this experiment was to characterise the polymorphisms of intron 1 of the interferon gammagene in Suffolk and Texel sheep, and to determine if the characterised haplotype variants were associated with faecal egg count variation in these breeds. Intron 1 haplotypes and faecal egg measurements were determined for Suffolks (n=113) and Texels (n=135). Four haplotypes were identified in Suffolks (A, B, C and D), two haplotypes as previously described, and two previously unknown haplotypes. However, there was no association between interferon gamma intron 1 haplotype and faecal egg count in Suffolks. In contrast, two previously identified haplotypes (A and B) were present in the Texel breed, and the B haplotype was associated with resistance to nematode infection (P=0.02). These results suggest that intron 1 of the interferon gamma gene has an important role in resistance to nematode infection in the Texel breed.     

Nitrogen and amino acid resorption in the small intestine of growing swine. 1. Animal study method, nitrogen content and amino acid composition of the small intestine chyme by administration of various proteins

The amount of endogenous N in the chyme at the end of the small intestine and the amino acid composition of the ileum chyme were ascertained with growing pigs with ileorectostomy after feeding qualitatively differentiated protein sources. In dependence on the protein used, distinct differences turned out for some amino acids with regard to their content in the ileum chyme. Bacteriologic and histologic investigations subsequent to the dissection of the test animals showed that - in comparison to the control animals-the functions of the small intestine remained the same during the time of the experiment. The experiment method described appears to be suitable for absorption investigations up to the end of the small intestine of growing pigs.     

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.     

Exploring polymorphisms and their effects on reproductive traits of the INHA and INHβA genes in three goat breeds

In this study, we report the analysis of INHA and INHβA gene polymorphisms in 786 goats of three breeds: Xinong Saanen (SN), Guanzhong (GZ) and Boer (BG). We identified three new allelic variants: P1–C80G and/126G (GenBank accession no. HQ202573) in the three goat breeds and P2–C936T (GenBank accession no. HQ202572) in SN and GZ goat breeds. At P1 locus, AA, AB and BB genotypes were found in the three goat breeds. At P2 locus, CC and CT genotypes were found in SN and GZ goat breeds. After comparing genotype distribution within the three goat breeds, BG had conspicuous differences from SN and GZ (P < 0.001) at P2 locus. The SNP locus was in Hardy–Weinberg disequilibrium at P1 locus in the three goat breeds (P < 0.05). At P2 locus, the SNP locus was in Hardy–Weinberg disequilibrium in SN and GZ goat breeds (P < 0.05). Association of polymorphisms with litter size was done at P1 locus in the three goat breeds. The result showed that AA genotype had remarkable litter size at P1 locus in the three goat breeds (P < 0.05). Therefore, these results suggest that INHA gene is a strong candidate gene that affects litter size in goats.     

Effect of the diets with pumpkin silage and synthetic β‐carotene on the carotenoid,immunoglobulin and bioactive protein content and fatty acid composition of colostrum

Forty Simmental cows were divided into four groups with 10 per group to investigate the effect of a transition diet supplemented with synthetic β‐carotene and natural carotenoid‐rich feed (pumpkin silage) on chemical composition of the colostrum. The control group (I) was fed grass and maize silage and supplemental concentrates, group II additionally received 400 mg/day/cow of synthetic β‐carotene, in group, III 40% DM of maize silage was replaced with pumpkin silage to increase β‐carotene intake by 400 mg in relation to group I, and in group IV 60% DM of maize silage was replaced with pumpkin silage without adjusting for β‐carotene. Colostrum was collected from the cows within 5 hr of calving to determine colostrum composition, the content of immunoglobulins and carotenoids, total antioxidant status (TAS), the content of some bioactive proteins and the composition of fatty acids. The study showed that the experimental diets had no effect on the gross composition of colostrum. The content of α‐carotene, β‐carotene and violaxanthin was significantly higher in group IV than in group I and that of lutein higher in group IV than in groups I and II. Total antioxidant status of colostrum in group IV was significantly higher than in group I. Groups III and IV were characterized by a higher concentration of immunoglobulin IgG in relation to group I. IgM level in groups II, III and IV was significantly higher than in group I and that in groups III and IV was also higher than in group II. The highest IgA concentration was observed in group IV and the lowest in group I. Lysozyme concentration was higher in group IV compared to groups I and II. The diets had no effect on the total content of SFA, MUFA and PUFA in colostrum.     

Influence of conjugated linoleic acids and vitamin E on milk fatty acid composition and concentrations of vitamin A and α‐tocopherol in blood and milk of dairy cows

The objective of this trial was to investigate the influences of conjugated linoleic acid (CLA ) and vitamin E (Vit. E) and their interactions on fatty acid composition and vitamins in milk (α‐tocopherol, retinol and β‐carotene) as well as on α‐tocopherol in blood of pluriparous cows from week 6 ante partum until week 10 post‐partum (p.p.). We assigned 59 pluriparous German Holstein cows to four treatment groups with the treatment factors CLA and Vit. E at two levels in a 2 × 2 factorial design. Milk fatty acid composition and milk vitamins were analysed on lactation days 7 and 28. α‐tocopherol in blood serum was analysed on days ?42, ?7, 1, 7, 14, 28 and 70 relative to parturition. Milk concentration of α‐tocopherol was influenced by Vit. E (p  < .001) and CLA (p  = .034). Percentage of cis ‐9, trans ‐11 CLA in total milk fat was influenced by treatment with CLA (p  < .001), while for percentage of trans‐ 10, cis ‐12 CLA an interaction between treatment and day (p  = .019), driven by an increase in both CLA groups from day 7 to day 28, was found. Serum ratios of α‐tocopherol to cholesterol were influenced by Vit. E (p  < .001). Results suggest that treatment with CLA during late pregnancy and early lactation is suitable to enhance the proportion of trans‐ 10, cis ‐12 CLA in milk and thereby influencing nutritional properties. As treatment with Vit. E did not have an impact on milk fatty acid composition, it might be possible to increase the antioxidative capacity of the dairy cow without affecting milk properties. Consequently, combined treatment with CLA and Vit. E might elicit synergistic effects on the cow and milk quality by increasing the proportion of CLA in milk fat as well as the excretion of Vit. E and the Vit. E levels in serum.     

β-defensins gene expression in tissues of the crossbred and Tibetan pigs

Delayed resumption of postpartum oestrous cycles in dairy cows is linked to low plasma insulin status and associated changes in ovarian function. Previous studies showed that plasma insulin can be raised by increasing dietary starch concentration, but it is not known if rumen digestible starch and bypass starch would be equally effective. The objective of the two experiments reported here was to investigate whether site of dietary starch digestion influences metabolic hormones and ovarian function. Cows were fed on a standard diet from calving until 40 days postpartum and then fed on treatment diets until 70 days postpartum. In Experiment 1, six cows were transferred to each of five diets with a starch content of 190 g/kg DM, of which 0.40, 0.45, 0.50, 0.55 or 0.60 was bypass starch. In Experiment 2, eight cows were transferred to each of four diets containing concentrates with low (L) or high (H) starch content and predominantly grass (G) or maize (M) silages in a factorial design. Between 60 and 70 days postpartum, plasma insulin was not influenced by site of starch digestion in Experiment 1, but insulin was increased by H (P = 0.020) and M (P = 0.048) in Experiment 2. Dietary bypass starch content negatively influenced plasma glucagon concentration (P = 0.046) and the insulin to glucagon ratio (P = 0.005) in Experiment 1. Growth hormone, insulin-like growth factor-I and leptin did not vary among diets in either experiment. The number of small (< 5 mm) ovarian follicles before ovulation tended (P = 0.056) to decrease with increasing dietary bypass starch content and was negatively related (P = 0.046) to plasma glucagon in Experiment 1, but there was no other dietary effect on ovarian function in either experiment. The main conclusion from these experiments is that rumen digestible starch and rumen bypass starch can be equally effective for maintaining plasma insulin and ovarian function of high-yielding dairy cows in early lactation.