理囊散治疗卵泡囊肿不孕奶牛血清白介素与激素的相关性分析

探讨理囊散治疗卵泡囊肿不孕奶牛血液生殖激素和白介素含量关系,分析其相关性与回归方程.用ELISA法测定理囊散治疗卵泡囊肿不孕奶牛前后血清中促卵泡激素（FSH）、促黄体激素（LH）、孕激素（P）和雌二醇（E2）的浓度以及血清IL-1、IL-6含量,统计分析了4种激素和两种白介素间的相关关系与回归方程.结果表明,IL-1、IL-6与P呈正相关,与FSH、LH、E2为负相关.提示卵泡囊肿不孕奶牛血清中白介素与激素的分泌存在一定的相关性.     

Milk protein polymorphisms in cattle (Bos indicus), mithun (Bos frontalis) and yak (Bos grunniens) breeds and their hybrids indigenous to Bhutan

In the current study, milk protein variation was examined in cattle (Bos indicus), mithun (Bos frontalis), yak (Bos grunniens) and their hybrid populations in Bhutan to estimate genetic variability, conduct genetic characterization and assess the possibility of gene flow between mithun and cattle. Isoelectric focusing of 372 milk samples from 11 populations detected four molecular types of β‐lactoglobulin (A, B, E and M), five molecular types of α_S1‐casein (A, B, C, E and X) and three molecular types of k‐casein (A, B and X). Mithun and yak shared alleles but were found to exhibit different allele frequencies for the proteins studied. The degree of genetic variability within populations was measured by average heterozygosity and ranged from 24–40% in cattle, 26% for yak and 33% for mithun. We also resolved the traditional mithun and cattle hybridization system via principal component analysis. Our results suggested secondary introgression of mithun genes to the village Thrabum population, and a close genetic relationship between Bhutanese indigenous cattle and Indian cattle.     

Expression and regulation of Enpp2 in rat uterus during the estrous cycle

Ectonucleotide pyrophosphatase/phosphodiestrase 2 (Enpp2) isolated from the supernatant of human melanoma cells is a lysophospholipase D that transforms lysophosphatidylcholine into lysophospatidic acid. Although multiple analyses have investigated the function of Enpp2 in the hypothalamus, its role in the uterus during the estrous cycle is not well understood. In the present study, rat uterine Enpp2 was analyzed by RT-PCR, Western blotting, and immunohistochemistry. Quantitative PCR analysis demonstrated that uterine Enpp2 mRNA was decreased during estrus compared to proestrus and diestrus. To determine whether uterine Enpp2 expression is affected by sex steroid hormones, immature rats were treated with 17β-estradiol (E2), progesterone, or both on postnatal days 14 to 16. Interestingly, the expression of Enpp2 mRNA and protein were down-regulated by E2 in the uterus during estrus but not during proestrus or diestrus, suggesting that Enpp2 may play a role in uterine function during estrus. Enpp2 is primarily localized in the stromal cells of the endometrium during proestrus and estrus. During diestrus, Enpp2 was highly expressed in the epithelial cells of the endometrium. Taken together, these results suggest that uterine Enpp2 may be regulated by E2 and plays a role in reproductive functions during female rat development.     

The localization and expression of epidermal growth factor and epidermal growth factor receptor in bovine ovary during oestrous cycle

Epidermal growth factor (EGF) is one of the important regulatory factors of EGF family. EGF has been indicated to effectively inhibit the apoptosis of follicular cells, to promote the proliferation of granulosa cells and the maturation of oocytes, and to induce ovulation process via binding to epidermal growth factor receptor (EGFR). However, little is known about the distribution and expression of EGF and EGFR in cattle ovary especially during oestrous cycle. In this study, the localization and expression rule of EGF and EGFR in cattle ovaries of follicular phase and luteal phase at different time points in oestrous cycle were investigated by using IHC and real-time qPCR. The results showed that EGF and EGFR in cattle ovary were mainly expressed in granulosa cells, cumulus cells, oocytes, zona pellucida, follicular fluid and theca folliculi externa of follicles. The protein and mRNA expression of EGF/EGFR in follicles changed regularly with the follicular growth wave both in follicular and in luteal phase ovaries. In follicular phase ovaries, the protein expression of EGF and EGFR was higher in antral follicles than that of those in other follicles during follicular growth stage, and the mRNA expression of EGFR was also increased in stage of dominant follicle selection. However, in luteal phase ovaries, the growth of follicles was impeded during corpus luteum development under the action of progesterone secreted by granular lutein cell. The mRNA and protein expressions of EGF and EGFR in ovarian follicles during oestrous cycle indicate that they play a role in promoting follicular development in follicular growth waves and mediating the selection process of dominant follicles.     

Rumen parameters of yaks (Bos grunniens) and indigenous cattle (Bos taurus) grazing on the Qinghai‐Tibetan Plateau

Yaks and indigenous Qaidam cattle and cattle‐yak crosses (C × Y) graze on the Qinghai‐Tibetan Plateau (QTP) throughout the year, but yaks are raised at higher elevations than cattle. Yaks do not receive supplementary feed whereas cattle require supplementary feed during harsh winter. We hypothesized that yaks would cope with the severe conditions of the QTP better than cattle and utilize the pasture more efficiently. We also hypothesized that differences between species would be pronounced in winter, when conditions are particularly harsh. To test these hypotheses, seasonal rumen fluid parameters of yaks, C × Y and cattle (n = 3 for each) were examined. Rumen fluid was collected in summer and winter from each genotype 2, 5 and 12 hr after a day of grazing. Concentrations of total volatile fatty acid (VFA), acetate, propionate, isobutyrate and isovalerate were greater in yaks than in cattle in summer (p < 0.05), while propionate concentration was lower in yaks than in cattle in winter (p < 0.05). Concentrations of ammonia and urea were greater (p < 0.001) in yaks than in cattle (p < 0.001) in summer, whereas, concentrations of free amino acids (AA) were greater in cattle than in yaks in summer and winter (p < 0.001). Concentrations of total VFA, acetate, propionate and butyrate decreased linearly (p < 0.05), whereas concentrations of isobutyrate and isovalerate increased linearly for yak and C × Y with sampling time (p < 0.05) in summer. In summer, concentrations of isobutyrate and isovalerate were greater in yaks than in cattle (p < 0.05). In conclusion, rumen fermentation characteristics of yaks showed that they coped better than cattle or C × Y in the harsh climate as we hypothesized. However, in contrast to our hypothesis, this emerged only in summer, when pasture was plentiful and not in winter, when pasture was scarce.     

Effects of Feeding Maize Stover and Cottonseed Cake on Onset of Puberty in Bunaji (Bos indicus) Heifers

A study was conducted to determine the influence of maize stover and cottonseed cake supplementation on age and live weight at onset of puberty in Bunaji heifers. A total of 49 pre-pubertal Bunaji heifers approximately 12 months of age were allotted randomly to one of two treatments of feed supplementation: (i) supplementation with maize stover and (ii) supplementation with cottonseed cake. The heifers were monitored for a period of 18 months for first observable oestrus or onset of puberty. The maize stover group were given maize stover (crop residue) supplementation ad libitum during the dry season; in the cottonseed cake group each heifer received 500 g of cottonseed cake per day during the dry season. The heifers supplemented with cottonseed cake attained puberty at an earlier age (23.9+/-1.2 months) than the heifers supplemented with maize stover (28.2+/-1.3 months; p<0.05). Similarly, heifers on the cottonseed cake supplementation attained puberty at a live weight of 270.5+/-3.5 kg; while the heifers on maize stover attained puberty at 237.6+/-5.8 kg (p <0.05). The cottonseed cake-supplemented heifers had higher daily gains than the maize stover-supplemented heifers. It is concluded that provision of cottonseed cake as a supplement for heifers hastened the onset of puberty when compared to the maize stover supplemented heifers. Puberty can be achieved at an early age with improved nutrition and this can in turn reduce the generation interval and thus increase the rate of genetic gain of indigenous animals. Provision of protein supplements in cattle is recommended, especially in the tropics where marked seasonality of rainfall results in extremely poor pastures and herbage and low available protein.     

Comparison of fattening performance in Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis) fed on high roughage diet

The objective of this study was to compare the fattening performance in Brahman grade cattle (crossbred cattle) and crossbred water buffalo at the same young age and fed with high roughage based fattening rations in the Philippines. Ten crossbred cattle and 10 crossbred water buffalo, aged between 18 and 24 months old were used in this experiment. The animals were fed diets consisting of 85% Napier or Para grass and 15% concentrate mixture (CM) on a dry matter (DM) basis. The grass, total DM intake and bodyweight gain were significantly (P < 0.01) higher for the crossbred water buffalo than for the cattle. There was no species significant difference in the digestion coefficient and feed conversion rate between the crossbred cattle and water buffalo. The return over feed cost for fattening was significantly (P < 0.05) higher in the crossbred water buffalo than in the cattle. These results clearly indicate that under high roughage‐based fattening rations, young crossbred water buffalo are better able to utilize the roughage and they perform better in terms of feed intake and live weight gains than the crossbred cattle in the Philippines.     

Melatonin and CIDR improved the follicular and luteal haemodynamics,uterine and ovarian arteries vascular perfusion,ovarian hormones and nitric oxide in cyclic cows

This study hypothesizes that melatonin with exogenous progesterone (CIDR) can improve follicular, luteal, ovarian and uterine haemodynamic of heat-stressed cows. Holstein cows (N = 12) studied for two spontaneous oestrous cycles during winter then divided equally during summer into the CIDR group received CIDR for 7 days and the melatonin group (Mel) received three injections of melatonin (75 mg/head) at the CIDR insertion, removal and ovulation days. Blood samples were collected to assay oestradiol (E2), progesterone (P4) and nitric oxide (NO). On day 0 (Ovulation), Mel had more small follicles (p < .05), higher ipsilateral and contralateral ovarian arteries (Ov.A.) peak systolic velocity (PSV), higher ipsilateral uterine artery (Ut.A.) PSV (p = .031) and blood flow volume (BFV), also Mel elevated contralateral Ut.A. PSV and BFV (p < .0001) but lowered contra Ut.A. pulsatility index (PI, p < .0001), E2 (p < .01) and NO (p < .0001). Mel increased the corpus luteum diameter (CL, p < .001), coloured area (p < .007) and P4 (p < .0001) on day 5 and reduced them (p < .05; p < .01) on Day 14. On day 10, Mel obtained CL diameter (p < .03) and coloured area (p < .002) of spontaneous that was higher than CIDR and decreased P4 (p < .003). Mel increased CL diameter, area and coloured area and decreased them thereafter. Mel increased the ipsilateral ovarian and uterine arteries PSV and BFV before ovulation and until day 8. Mel increased P4 and decreased NO until days 6 and 14. In conclusion, the improvement in follicular, luteal, ovarian and uterine haemodynamic and the decrease of NO production proved our hypothesis Melatonin doses higher than 75 mg/head is recommended to improve the heat-stressed cow's fertility.     

Effects of Calf Removal at Parturition on Postpartum Ovarian Activity in Zebu (Bos indicus) Cows in the Humid Tropics

To assess endocrine and morphological responses of ovaries to total weaning at parturition, 6 Zebu (Bos indicus) cows 5 years or older were investigated. Following parturition, blood samples were collected daily during the first month and twice weekly thereafter until day 60 to determine concentrations of progesterone (P₄) and prostaglandin F_2α metabolite.It took between 25 to 32 days to complete uterine involution.The prostaglandin metabolite remained elevated for a mean period of 14.2 days (range, 4-21) postpartum. Five of the animals resumed cyclicity with a short estrous cycle starting between days 7 to 34 and lasting between 7 and 14 days. No estrous behavior was recorded prior to the short estrous cycles, but subsequent normal-length estrous cycles were all preceded by signs of estrus. In the 1 animal that resumed cyclicity with an estrous cycle of normal length on day 37 (length 20 days), the cycle was preceded by estrous behavior.Progesterone concentrations reached a mean maximum of 4.8 nmol liter⁻¹ during the short estrous cycles, and prostaglandin metabolite concentrations peaked while P₄ concentrations were decreasing. P₄ concentrations reached a mean maximum of 12.2 nmol liter⁻¹ during the estrous cycles of normal length. The interval from parturition to the first estrous cycle of normal length varied between 16 and 48 days, and the length of the cycle was 18 to 22 days.Starting 2 days postpartum, ovaries from 5 of the cows were scanned by ultrasonography every second day until day 30 postpartum. Medium-sized follicles were detected between days 4 to 7 postpartum in 4 of the scanned cows that later had short estrous cycles. The time between parturition and the appearance of the first dominant follicle was 7.6 days (range 6-10 days). The interval between parturition and the appearance of the first ovulatory-sized follicle was 10.2 days (range 8-13 days). In 3 of the scanned cows this ovulatory-sized follicle ovulated.We conclude that cyclic ovarian activity in Zebu cows can start early in the postpartum period in the absence of offspring, and that short luteal phases, not preceded by estrous behavior, may play an important role in establishing normal postpartum ovarian activity.     

Plasma hormones and metabolites in cattle in relation to breed (Belgian Blue vs Holstein) and conformation (double-muscled vs dual-purpose type)

Four Belgian Blue double-muscled type (BBDM) bulls, four Belgian Blue dual-purpose type (BBDP) bulls and four Holstein bulls were used in a fattening trial in order to relate differences in the extent of muscle development and adiposity to differences in digestibility, endocrine status, protein and lipid metabolism. The larger muscle development in BBDM animals was associated with a trend to higher nitrogen retention, higher food conversion efficiency (p<0.05) and lower apparent digestibility (p<0.05). No difference was found between the groups for plasma glucose concentration. Higher creatinine, lower -amino nitrogen, lower triglycerides and higher non-esterified fatty acid plasma levels were observed in BBDM as compared to Holstein bulls (p<0.05), the BBDP group being intermediate. A trend to a higher cholesterol plasma level was found in BBDM animals. There was no difference between the three groups in plasma fatty acid composition, except for the C_14:0 content. Some of the differences in plasma metabolites were related to carcass composition and endocrine regulation, a decrease in muscle development and an increase in adiposity being associated with lower growth hormone production (p<0.05) and higher insulin (p<0.05) and IGF₁ secretions. The insulin/growth hormone ratio at the end of the fattening period was 0.0011, 0.0018 and 0.0069 in BBDM, BBDP and Holstein bulls, respectively, and was directly associated with fat deposition.Abbreviations BBDM Belgian Blue breed double-muscled type - BBDP Belgian Blue breed dual-purpose type - IGF₁ insulin-like growth factor 1 - NEFA non-esterified fatty acids - T₃ triiodothyronin - T₄ tetraiodothyronin     

牦牛CRH基因克隆及其在生殖轴中的表达研究

为了探讨促肾上腺皮质激素释放激素（corticotropin-releasing hormone,CRH）基因在牦牛繁殖中的调控作用,试验分别采集5头成年母牦牛和5头成年母黄牛的下丘脑、脑垂体前叶、输卵管、卵巢和子宫等组织,通过RT-PCR技术及生物信息学软件对牦牛CRH基因编码区进行扩增、克隆及序列分析,并构建系统进化树;采用实时荧光定量PCR法检测CRH基因的组织表达。结果表明,牦牛CRH基因编码区全长573 bp,编码190个氨基酸。牦牛与黄牛、猫、鸡、人、小鼠、大鼠和猪的CRH基因核苷酸同源性分别为99.8%、43.3%、36.1%、46.2%、39.0%、38.5%和56.4%。系统进化树表明,牦牛与黄牛亲缘关系最近,并与其他物种类聚,符合物种进化规律。CRH蛋白分子式为C₉₂₀H₁₅₀₃N₂₇₉O₂₆₃S₃,分子质量为20776.95 u,理论等电点为10.95,其氨基酸组成中亮氨酸（L）、脯氨酸（P）、丙氨酸（A）和精氨酸（R）所占比例较高,分别为15.8%、12.6%、12.1%和10.5%。CRH蛋白为不稳定亲水蛋白,存在信号肽和跨膜结构。CRH蛋白二级结构中α-螺旋、β-转角、延伸链和无规则卷曲分别占41.05%、1.05%、8.42%和49.48%,三级结构分析结果与其相一致。CRH基因mRNA在牦牛下丘脑中表达量最高,显著高于其他组织（P<0.05）,在子宫中表达量最低。牦牛脑垂体前叶CRH基因mRNA表达量极显著高于黄牛（P<0.01）,在卵巢、输卵管中表达量显著高于黄牛（P<0.05）,而两个品种在下丘脑、子宫中的表达量差异不显著（P>0.05）。提示CRH基因在牦牛繁殖调控中具有重要作用。     

Gene and haplotype polymorphisms of the Prion gene (PRNP) in Japanese Brown, Japanese native and Holstein cattle

Polymorphisms in the prion protein gene ( PRNP ) are known to be associated with transmissible spongiform encephalopathies in human, sheep and goats. There is tentative association between PRNP promoter polymorphism and bovine spongiform encephalopathy (BSE) susceptibility in cattle. In this study, we genotyped for six bovine PRNP polymorphic sites including a 23-bp indel in the promoter, a 12-bp indel in the intron 1, two nonsynonymous single nucleotide polymorphisms (SNPs), octapeptide repeats in the coding region and a 14-bp indel in the 3'-untranslated region in 178 animals representing Japanese Brown, Kuchinoshima feral, Mishima, Japanese Shorthorn and Holstein. In 64 Japanese Brown cattle, three indel sites were polymorphic. All of the six sites were monomorphic in Kuchinoshima. The 23-bp and 12-bp indel sites were polymorphic in Mishima cattle. The 23-bp and 14-bp indel sites were polymorphic in Japanese Shorthorn cattle. Both SNP sites were monomorphic in all cattle examined in this study. At the 23-bp indel site, the genotype frequencies of Japanese Brown and Holstein breeds were similar to that of BSE affected cattle. We estimated 12 different haplotypes from these genotypic data. A '23-12-K6S14+' haplotype was the major haplotype in all populations, whose frequencies ranged from 0.50 to 1.00.     

Quantitative analyses of insulin-like peptide 3 and sex steroid hormones in dominant follicles and corpora lutea during the estrus cycle and in follicular cysts in beef heifers

Insulin-like peptide 3 (INSL3) and sex steroids were measured in bovine dominant follicles and corpora lutea during the estrus cycle and in follicular cysts. Paired ovaries from beef heifers (n = 47) were classified, by their morphological features, either into four stages of the estrus cycle (Day 1 = day of ovulation, Day 20 = day of estrus) as Stage I (Days 1–4; n = 8), Stage II (Days 5–10; n = 10), Stage III (Days 11–17; n = 8), and Stage IV (Days 18–20; n = 11) or follicular cystic (n = 10). Cysts (n = 15) were subdivided into estrogen-active (n = 7) and estrogen-inactive (n = 8) cysts. INSL3, testosterone, and estradiol-17β concentrations in the dominant follicular fluid of Stage IV were higher than those in Stages II and III (P < 0.05). INSL3 concentrations in the cystic fluid were similar to those in dominant follicles at Stage IV, whereas testosterone and estradiol-17β concentrations were lower in cysts (P < 0.05). INSL3 content per estrogen-inactive cyst was higher than that of Stage IV (P < 0.05). INSL3 and progesterone concentrations in luteal tissue and contents per corpus luteum were higher in Stages II and III (P < 0.05). In conclusion, INSL3 secretion in bovine dominant follicles increased with maturation. Follicular cysts may retain the production of INSL3 during their formation but tend to lose the capacity for testosterone secretion. Estrogen-inactive cysts subjected to advanced atresia may accumulate more INSL3. INSL3 production in bovine corpora lutea is enhanced during maturation.     

Relationship between peripheral plasma inhibin and FSH concentrations in Sahiwal cows (<Emphasis Type="Italic">Bos indicus</Emphasis>) and Murrah buffaloes (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>) during estrous cycle

The present investigation was undertaken to study the relationship between circulating inhibin and FSH concentrations during the estrous cycle in buffaloes and Sahiwal cattle. The pattern of inhibin concentrations was similar, with peak concentrations on Day -2 (Day 0 = day of estrus) and minimum concentrations on Days 12 and 11 in buffaloes and cattle, respectively. Circulating FSH concentrations were the highest on Day 0 and lowest on Days 8 and 13 in buffaloes and cattle, respectively. Peripheral plasma inhibin concentrations were negatively correlated to FSH concentrations in buffaloes (r = -0.27, P < 0.01) and cattle (r = -0.35, P < 0.01) indicating that inhibin is involved in negative feedback regulation of FSH in both these species.     

Questionnaire investigation to clarify the occurrence rate and characteristics of maternal rejection behavior in Japanese black cattle (Bos taurus)

We conducted a questionnaire investigation among breeding farmers to clarify the actual conditions of maternal rejection in Japanese Black cattle. We asked keeping experience of maternal rejective cows and compared occurrence patterns, rejective behavior manners, birth assistance methods, colostrum feeding method for calves, parity and rearing conditions of the cows. We found that 24% of the farms had kept rejective cows and 6% of the cows in these farms indicated maternal rejections. The most common occurrence pattern was ‘Occurred from the first birth (65.6%)’ and behavior manner was performing no maternal grooming with aggressive behavior (75%). Almost all the farmers assisted in each parturition (P < 0.05). There was not a significant difference among the colostrum feeding methods, but a significant difference in the rearing conditions of the cows, except for the cows with no data (P < 0.005). The parity of the rejective cows did not significantly differ in association with rejective behavior manners, birth assistance methods, colostrum feeding methods or rearing conditions. These results suggest that the occurrence rate of maternal rejection in Japanese Black cattle was approximately 6% and many of the rejective cows continuously performed no maternal grooming with aggressive behavior.