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1.
基于蓝舌病（bluetongue,BT）的危害及RNA阳性质控品研制的重要性,试验将pTrcHis-MS2质粒借助点突变技术在噬菌体外壳蛋白15和16氨基酸之间插入组氨酸（His）蛋白标签,构建了pTrcMS载体。将pMD19-T-BTV质粒与pTrcMS质粒分别进行KpnⅠ和Hind Ⅲ双酶切,然后连接构建pTrcMS-BTV重组质粒。将pTrcMS-BTV重组菌进行原核表达,产物借助偶联His蛋白标签的磁珠捕获技术获得纯化的蓝舌病假病毒颗粒,进行特性鉴定并定量。结果显示,研制的pTrcMS-BTV物质经PCR方法评价纯度高,无基因DNA污染;透射电镜观察形态为不规则的直径约为26 nm的多边形;RT-PCR方法检测稳定性表明该物质耐RNase A,易保存;应用荧光RT-PCR检测表明该物质溶液最低检测限可达到2.5×102拷贝/mL。蓝舌病假病毒物质将为蓝舌病分子生物学检测提供阳性质控物质。 相似文献
2.
Site of protein digestion in steers fed sorghum grain diets. I. Effect of level of feed intake 总被引：1，自引：0，他引：1
S H Rahnema B Theurer J A Garcia M C Young R S Swingle 《Journal of animal science》1987,64(5):1533-1540
Two digestion trials were conducted with seven crossbred, abomasally cannulated yearling steers (400 kg) to study the effect of level of feed intake on the site and extent of feed and microbial protein digestion. Steers, in a crossover design experiment, were fed an 81.5% steam-processed flaked (SPF) sorghum grain diet at either 95% (four steers) or 75% (three steers) of their ad libitum intakes. At the end of the first trial, steers were switched to the opposite treatment. Dysprosium (31 to 32 micrograms/g) was used as an external marker. Feed, abomasal contents and fecal grab samples were taken at 12-h intervals advancing by 2 h each day over a 6-d total fecal collection period. Organic matter (OM) intakes were 6,102 and 4,570 g for the two treatments. Higher level of intake increased (P less than .05) quantities of OM, crude protein and trichloroacetic acid precipitable protein entering the small intestine, digested post-ruminally and digested in the total tract. The higher level of intake decreased (P less than .05) the percentage of bacterial protein (BP) present in the abomasum and percent post-ruminal BP digestion; however, the amounts of BP and non-BP entering the small intestine and digested post-ruminally were greater (P less than .05) in steers fed 95% ad libitum. Most of the feed protein was degraded in the rumen with both treatments. Predicted true feed protein digestibilities were 91.1 and 91.7% for 95 and 75% of ad libitum intakes.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
3.
M S Kerley A R Kinser J L Firkins G C Fahey L L Berger 《Journal of animal science》1985,61(2):504-513
An experiment examining nutritional effects of dietary corncob particle size was conducted using ruminal-, duodenal- and ileal-cannulated sheep in a 4 X 4 Latin square design. Site of nutrient digestion and digesta flow were the principal criteria evaluated. Analyses of dry matter (DM), N, starch and neutral detergent fiber (NDF) were performed on feed, feces and digesta samples. Chromic oxide-impregnated paper was used as an external marker to estimate digestibilities at different sites along the gastrointestinal tract. Ruminal pH and volatile fatty acid molar proportions were also determined. All diets (74.9% concentrates: 25.1% corncobs) were pelleted and were similar in ingredient composition but varied in corncob particle size (corncob mean particle sizes were 6.5, 5.4, 1.4 or .8 mm). Dietary crude protein levels differed little among treatments. Starch concentration was higher in diets containing the larger corncob particles while NDF, acid detergent fiber (ADF), cellulose and hemicellulose concentrations were lower in diets containing larger particles, suggesting a reaction between starch and fiber moieties during the pelleting process. Starch flow past the duodenum decreased (P less than .05) as dietary corncob particle size decreased. Apparent NDF digestion before the duodenum was highest for sheep fed diets containing 1.4-mm corncobs (P less than .05). Apparent starch digestion in the small intestine decreased (P less than .05) as dietary corncob particle size decreased. A considerable amount of NDF was apparently digested in the small intestine of sheep consuming diets containing 5.4- and .8-mm corncobs. Likewise, a substantial amount of NDF was apparently digested in the large intestine. Few differences in apparent total tract digestibilities were noted.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
4.
L Rice E A Ott D K Beede C J Wilcox E L Johnson S Lieb P Borum 《Journal of animal science》1992,70(4):1175-1181
Oral tolerance tests were performed on 13 neonatal foals to determine their ability to digest disaccharides on d 1, 3 and 5 postpartum. Foals were assigned randomly to treatments consisting of 20% (wt/vol) solutions of either maltose, lactose, or sucrose, dosed at 1 g/kg of BW, or glucose, dosed at .5 g/kg of BW. After a 2-h fast, an initial blood sample was collected via jugular catheter. Foals were administered the appropriate solution orally, and blood was collected every 15 min for 1 h and then every 30 min for 3 h. Plasma glucose increased after dosing with lactose or glucose but not with sucrose. Plasma glucose concentrations increased slightly on d 3 and 5 in foals dosed with maltose. These findings suggest that although lactose is well digested by neonatal foals, maltose is digested only slightly, and sucrose is not digested by d 5. Results of this experiment indicate that maltose and sucrose would not be suitable for inclusion in artificial diets for foals less than 1 wk old. Oral tolerance tests could be useful for determining the ability of premature or sick foals with lactose intolerance to digest alternate carbohydrate sources. 相似文献
5.
伪狂犬病毒EP0基因的克隆及其在大肠杆菌中的表达研究 总被引：1，自引：1，他引：0
为了研究EP0蛋白在伪狂犬病毒粒子中的定位及其功能,试验利用PCR从伪狂犬病毒基因组中扩增PRV EP0基因的编码区,克隆至pMD18-T载体,构建重组质粒pMD-EP0。经测序鉴定正确后,用EcoRⅠ和Hind Ⅲ双酶切pMD-EP0,回收EP0基因与经同样双酶切处理的pET-32a(+)进行连接,构建重组质粒pET-EP0。将pET-EP0转化大肠杆菌BL21(DE3),经IPTG诱导后,通过SDS-PAGE和Western blotting检测融合蛋白的表达情况及反应原性。结果表明,成功扩增得到EP0基因编码区序列,大小为1227 bp;重组表达质粒 pET-EP0经诱导后,EP0融合蛋白获得了表达,大小约为75 ku,主要以包涵体形式存在,且能与伪狂犬病毒阳性血清反应。 相似文献
6.
Deoxyribonucleic acid (DNA) was extracted from bovine herpesvirus type 1 (BHV-1) isolated from a stillborn porcine fetus, from the Cooper reference strain of BHV-1, and from an Ontario bovine respiratory isolate. Each DNA was digested with the restriction endonucleases HindIII, EcoRI, HpaI and BamHI. Except for very minor differences in the patterns produced after digestion with EcoRI and HindIII, the DNA of the porcine isolate reacted in a similar manner to the bovine viruses, and it was concluded that the porcine virus is genetically similar to bovine isolates of BHV-1. 相似文献
7.
Forty-two samples were taken from the contents of the proximal small intestine of two lactating dairy cows fitted with re-entrant duodenal cannula. Most samples were free of detectable amylase activity. The (chymo)trypsinogen present was only partially activated to (chymo)trypsin. The activation was continued in vitro: slowly at the original pH of the samples (between pH 2.8 and 4.2), and faster after neutralization or a slight alkalinization. The effect of Ca, EDTA and soybean inhibitor on the activation of trypsinogen was also studied. The pancreatic enzymes were inactive in the acid pH range of the samples, but pepsin was markedly active. At pH 3.8 casein was digested rapidly by purified pepsin and slowly by the samples (agar-plate experiments). In model experiments performed with purified enzymes, pepsin digested (chymo)trypsin rapidly at pH 1-2 and slowly at pH 3.8. In the intestinal juice (chymo)trypsin and their zymogens seemed to be unaffected by pepsin under the conditions of the samples. It is concluded that the conditions prevailing in the duodenum/upper jejunum of the experimental cows account for a gastric-type digestion, despite the presence of pancreatic enzymes. In vivo the intestinal contents pass in distal direction. Meanwhile the pH of the chyme gradually increases and gives rise to first an increase of enterokinase activity accounting for a faster activation of the zymogens; second a start of function of activated pancreatic proteases and third a gradual decrease of pepsin activity and finally to its irreversible denaturation. Thus the development of intestinal type digestion is delayed in ruminants. 相似文献
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A micromethod was used in order to quantitate intestinal D-xylose absorption in young and extremely small birds. This test was performed in broilers collected from two farms from which birds were extremely uneven in body size and were passing poorly digested or undigested feed. A similar syndrome had been seen on all grow-outs during the 6 months before this investigation. Broilers were also collected and tested from three farms where no clinical signs of disease were seen. D-Xylose absorption peaks and curves for normal broilers closely resembled those observed in normal humans. Mean plasma D-xylose concentrations for virus-infected broilers and for broilers with pale-bird syndrome were consistently lower than concentrations for normal broilers (P = 0.009). Reoviruses, small coronavirus-like particles, small round virus particles, and abundant bacterial flagellar fragments were seen in fecal samples from broilers with pale-bird syndrome. Production performance was lowest on farms showing clinical signs of this syndrome. 相似文献
10.
S Tola G Idini A M Rocchigiani D Manunta P P Angioi S Rocca M Cocco G Leori 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1999,46(3):199-206
Mycoplasma agalactiae and Mycoplasma bovis are closely related species in phylogenetic terms. Pulsed Field Gel Electrophoresis (PFGE) of SmaI, EclXI, Bsi WI, MluI, BssHII, SalI, XhoI, NruI and ApaI digested DNAs were used to analyse and to compare restriction fragment length polymorphism between M. agalactiae and M. bovis and to estimate their genome sizes. SmaI, EclXI and Bsi WI enzymes cleaved DNAs of both microrganisms. MluI, BssHII, SalI, XhoI and NruI digested only M. agalactiae DNA whereas ApaI cut only M. bovis DNA. The total DNA length was established to be 945 +/- 8.4 Kb for M. agalactiae and 961 +/- 18.9 Kb for M. bovis. 相似文献
11.
以已克隆测序的家蚕微孢子虫 (镇江株 ,Nosemabombycis)的核糖体小亚单位RNA(SSUrRNA)编码基因(12 0 5bp)为模板 ,用随机引物合成法标记的探针 ,在与 9种微孢子虫及家蚕基因组DNA的 6种限制性内切酶的酶切产物的Southern杂交图谱中 ,9种微孢子虫基因组DNA酶切产物的杂交图谱极为相似 ,而与家蚕基因组DNA的任何一种酶的酶切产物均无杂交信号 ,表明微孢子虫和家蚕的SSUrRNA基因同源性很低或没有同源性。同时还证实了已克隆的SSUrRNA基因来源于微孢子虫基因组DNA ,不是从家蚕基因组DNA扩增而来。在酶解较为完全的酶切产物的杂交图谱中 ,微孢子虫基因组DNA中SSUrRNA基因的拷贝数至少在 15以上 相似文献
12.
The digestion of starch and fibre by adult cockerels fed peas which were whole, ground, autoclaved, heated, dehulled, cooked or supplemented with a cellulose degrading enzyme was studied. The starch in ground peas that had been autoclaved, heated or dehulled was slightly, though not significantly, better digested than the starch in peas that had been ground only. Feeding peas whole greatly reduced starch digestibility. Cooking failed to improve starch digestibility as retrograded starch produced in the cooking process was not digested. The more finely ground the foodstuff, the better the in vitro enzymatic hydrolysis of starch. Starch digestibility correlated well (r2 = 0.80) with the true metabolisable energy values obtained for the different pea treatments. In experiment 1 cockerels digested on average 0.22 of the pea fibre from the different forms of peas. A significant decrease in xylose digestion was observed when birds were fed dehulled peas, birds excreting more xylose than they ingested. In experiment 2, cockerels digested on average 0.38 of the pea fibre from different forms of peas. A reduction in fibre digestion was observed when birds were fed cooked peas as a consequence of a decreased digestibility of all monosaccharide residues. A slight increase in fibre digestion was observed when peas were augmented with 'cellulase', entirely because of an increase in xylose digestion. 相似文献
13.
不同组合饲粮对绵羊消化代谢的影响 总被引：5，自引：2，他引：3
选用3只安装永久瘤胃瘘管的甘肃高山细毛羯羊(1.5岁,平均体重22.5kg),采用3×3拉丁方设计,研究3种组合饲粮,即对照饲粮(Ⅰ)、含甜菜渣饲粮(Ⅱ)、含苹果渣饲粮(Ⅲ)对绵羊消化代谢的影响。试验分3期,每期预试期10d,正试期9d。结果表明,饲粮Ⅰ、Ⅱ的干物质(DM)和有机物质(OM)消化量与消化率均显著高于Ⅲ(P＜0.05);饲粮Ⅱ的中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)消化量及消化率呈现出高于Ⅰ、Ⅲ的趋势(P＞0.05),饲粮Ⅲ的NDF和ADF消化率也呈现出低于Ⅰ的趋势;饲粮Ⅱ的氮消化量显著高于Ⅰ(P＜0.05),其消化率显著高于饲粮Ⅰ、Ⅲ(P＜0.05),呈现出其氮存留量高于Ⅰ及氮存留率高于Ⅰ、Ⅲ的趋势;饲粮Ⅱ钙的表观消化率显著高于Ⅰ(P＜0.01)、Ⅲ(P＜0.05),Ⅲ也显著高于Ⅰ(P＜0.05);饲粮Ⅲ的磷表观消化量显著高于Ⅰ、Ⅱ(P＜0.05),且呈现饲粮Ⅲ磷消化率高于Ⅱ,Ⅱ也高于Ⅰ的趋势。 相似文献
14.
J. P. Lynch P. O'Kiely R. Murphy E. M. Doyle 《Journal of animal physiology and animal nutrition》2014,98(4):731-738
Maize stover (total stem and leaves) is not considered a ruminant feed of high nutritive value. Therefore, an improvement in its digestibility may increase the viability of total forage maize production systems in marginal growth regions. The objective of this study was to describe the changes in chemical composition during the storage of contrasting components of maize stover (leaf, upper stem and lower stem) treated with either of two lignin degrading white‐rot fungi (WRF; Pleurotus ostreatus, Trametes versicolor). Three components of maize stover (leaf, upper stem and lower stem), harvested at a conventional maturity for silage production, were digested with either of two WRF for one of four digestion durations (1–4 months). Samples taken prior to fungal inoculation were used to benchmark the changes that occurred. The degradation of acid detergent lignin was observed in all sample types digested with P. ostreatus; however, the loss of digestible substrate in all samples inoculated with P. ostreatus was high, and therefore, P. ostreatus‐digested samples had a lower dry matter digestibility than samples prior to inoculation. Similarly, T. veriscolor‐digested leaf underwent a non‐selective degradation of the rumen‐digestible components of fibre. The changes in chemical composition of leaf, upper stem and lower stem digested with either P. ostreatus or T. veriscolor were not beneficial to the feed value of the forage, and incurred high DM losses. 相似文献
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The objective was to evaluate the nutritive value of short-cutting cycle, high temperature-dried (SCCHTD) alfalfa compared to timothy hay. This was achieved by carrying out 4 x 4 Latin Square digestibility trial using 4 Thoroughbred (one three-quarter Thoroughbred) horses (mean liveweight, 531 kg). The four dietary treatments were 0AA (timothy hay only), 33AA (0.33 alfalfa: 0.67 timothy hay), 67AA (0.67 alfalfa: 0.33 timothy hay) and 100AA (alfalfa only). Digestibility data were obtained by using acid-insoluble ash to estimate apparent digestibility coefficients of nutrients. Rate of passage of the feedstuff was determined using chromium-mordanted hay. Plasma triglyceride and cholesterol concentrations were estimated. The digestibilities of organic matter (0.63), energy (0.57) and crude protein (0.74) of the alfalfa were significantly (P less than 0.001) higher than those for the hay (0.45, 0.43 and 0.36 respectively). The fibre components of alfalfa and hay were digested to the same extent but the ether extract of alfalfa was less well digested. Alfalfa saponins had no consistently significant effects on plasma cholesterol and triglyceride values but may contribute to the negative digestibility of alfalfa ether extract. We conclude that SCCHTD alfalfa is of much higher nutritive value than timothy hay when fed to Thoroughbred horses. 相似文献
17.
T. M. Storlien H. Volden T. Almøy K. A. Beauchemin T. A. McAllister O. M. Harstad 《Acta Agriculturae Scandinavica, Section A - Animal Sciences》2013,63(2):98-109
AbstractThe main objective of this study was to elucidate the potential for prediction of enteric methane (CH4) emissions from dairy cows by using predicted rumen plus hindgut digested (fermented nutrients) and total tract digested nutrients (by using NorFor) as input variables. Twenty-one experiments (78 dietary treatments) were collected. The data-set was used to develop prediction models and to test their and extant models ability to predict enteric CH4 emissions. Models were compared based on mean squared prediction error and concordance correlation coefficient (CCC) analysis. Fermented nutrients did not predict enteric CH4 emissions adequately (CCC < 0.420). Including total digested (td) nutrients in the model [CH4 (MJ/d) = ?2.13 + 1.64 tdOM (kg/d) ?9.74 tdFat (kg/d) + 1.64 tdNDF (kg/d)] predicted enteric CH4 emissions more precisely (CCC = 0.733), and showed an improvement in the prediction of enteric CH4 emissions over the extant models tested. 相似文献
18.
The effect of supplementation with different levels of cracked corn on the sites of OM, total dietary fiber (TDF), ADF, and starch digestion in steers fed fresh alfalfa indoors was determined. Six Angus steers (338 +/- 19 kg) fitted with cannulas in the rumen, duodenum, and ileum consumed 1) alfalfa (20.4% CP, 41.6% NDF) ad libitum (AALF); 2), 3), and 4) AALF supplemented (S) with .4, .8, or 1.2%, respectively, of BW of corn; or 5) alfalfa restricted at the average level of forage intake of S steers (RALF), in a 5 x 5 Latin square design. Total OM intake was lower (P < .01) in steers fed RALF than in those fed AALF but level of forage intake did not affect sites of OM, TDF, or starch digestion (P > .05). Forage OM intake decreased (P < .01) linearly (8,496 to 5,840 g/d) but total OM intake increased (P = .03) linearly (8,496 to 9,344 g/d) as corn increased from .4 to 1.2% BW. Ruminal apparent and true OM disappearance was not affected, but OM disappearing in the small intestine increased (P < .01) linearly with increasing levels of corn. Total tract OM digestibility (71.2 to 76.2%) and the proportion of OM intake that was digested in the small intestine (15.4 to 24.5%) increased (P < .01) linearly as corn increased. The TDF and ADF intakes decreased (P < .01) linearly as level of corn increased. Total tract TDF and ADF digestibilities were not different among treatments (average 62.9 and 57.8%, respectively). Starch intake and starch digested in the rumen and small and large intestine increased (P < .01) linearly with increasing corn level. Ruminal pH and VFA concentrations decreased and increased (P < .01), respectively, with increasing corn. Supplementation with corn increased OM intake, decreased forage OM intake, and increased the proportion of OM that was digested in the small intestine, but fiber digestion was not affected. 相似文献
19.
Kinetics of solutes and particles of different size in the digestive tract of cattle of 0.5–10 years of age,and relationships with methane production 
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下载免费PDF全文 F. Grandl A. Schwarm S. Ortmann M. Furger M. Kreuzer M. Clauss 《Journal of animal physiology and animal nutrition》2018,102(3):639-651
The digestive physiology of cattle is characterised by comparatively long digesta mean retention times (MRTs), a particle sorting mechanism (difference in MRTs of large vs. small particles) and a distinct digesta washing (difference in MRTs between particles and fluids) in the reticulorumen (RR). How these processes mature during ontogeny, and how they link to other digestion characteristics and methane production, is largely unknown. We used a set of passage markers (Co‐EDTA for fluids and hay particles of 2, 5 and 8 mm length mordanted with Cr, La and Ce, respectively) to measure MRTs in 12 heifers (0.5–2.1 years; hay only) and two groups of 15 lactating cows (2.4–10.0 years; forage‐only vs. forage‐concentrate diet). The MRTs differed between markers (Co < Cr < La < Ce) and were longer in heifers than cows, consistent with the lower feed intake in heifers. MRTs were mostly similar between cow groups and increased with age. Digesta washing was not affected by group, age, feed intake and number of chews per unit of feed. The degree of digesta washing was not related to CH4 measures. Particle sorting was more prominent in cows than heifers but did not differ between cow groups or change with age in cows. This could be the consequence of the abrupt increase in intake from heifers to cows at a time when gut capacity is not yet fully developed; particle sorting might then clear smaller particles from the RR sooner allowing a higher intake. Surprisingly, CH4 yield per ingested feed did not correlate with MRTs, and CH4 yield per unit of digested fibre decreased with increasing MRTs and with increasing fibre digestibility. As this pattern occurred in heifers and both cow groups, it appeared independent of age, indicating a mechanism that has not been described in the literature so far and requires further investigation. 相似文献
20.
Abnormal structure of the canine oncogene, related to the human c-yes-1 oncogene, in canine mammary tumor tissue. 总被引：1，自引：0，他引：1
N Miyoshi S Tateyama K Ogawa R Yamaguchi H Kuroda N Yasuda T Shimizu 《American journal of veterinary research》1991,52(12):2046-2049
Cellular oncogenes of genomic DNA in 6 canine primary mammary tumors were screened by Southern blot analysis, using 7 oncogene probes. A canine genomic oncogene related to the human c-yes-1 oncogene was detected as abnormal bands in solid carcinoma genomic DNA digested with EcoRI, HindIII, HindIII-EcoRI, or HindIII-BamHI. Comparison was made between other tumor specimens and control specimens obtained from 4 clinically normal dogs--1 mixed breed and 3 Shiba Inu dogs (the same breed as the dog from which the solid carcinoma was obtained). These abnormal bands were 0.1 to 1 kilobase shorter than the normal gene. However, digestion of genomic DNA obtained from normal WBC of this dog also produced all of the abnormal bands as observed in digested DNA from the solid carcinoma tissue. Therefore, in this dog, the genomic DNA of all somatic cells from the ontogenic stage still had the abnormal sequences related to the human c-yes-1 oncogene, and it is possible that this abnormal structure may have some role (eg, as an initiator) in tumorigenesis or the progression of this tumor. 相似文献