抗草甘膦转基因大豆及其加工产品的环介导等温扩增检测技术

本研究旨在建立抗草甘膦转基因大豆及其加工产品中外源基因的环介导等温扩增(loop-mediated isothermal amplification,LAMP)检测技术.根据转基因大豆外源CP4-EPSPS基因的6个区域设计4条特异性引物,利用LAMP法对选取的大豆、豆粕及含有大豆成分的饲料样品进行检测.结果提示,LA...     

转基因豆粕中内源基因和外源基因检测方法的建立

抗草甘膦(Glyphosate)大豆(商品名:Roundup Ready Soybean)是在传统大豆中转入外源基因CaMV-35S启动子、抗草甘膦基因CP4-EPSPS和NOS终止子而得到的[1].由大豆加工而成的豆粕经过高温浸提、脱溶等过程,其中的DNA成分遭到破坏,容易发生降解等现象.因而如何从豆粕中有效提取DNA成分,并进行可靠的检测就成为检测部门的重要问题.     

转基因成分在肉鸡体内的代谢残留及对生化指标、器官发育的影响

试验选择1日龄AA肉仔鸡720只,随机分成4个处理组,用含有27%(25%)3、6%(34%)的抗草甘膦转基因豆粕日粮和常规豆粕日粮饲喂肉鸡,通过检测其肝脏、肾脏、脾脏、胸肌、腿肌、十二指肠、空肠和回肠的转基因成分,探讨饲料中转基因成分在家禽体内的代谢残留状况,同时研究了抗草甘膦转基因豆粕对肉鸡血清生化指标和器官发育的影响,结果表明,以定性PCR检测的转基因豆粕日粮肉鸡的八种样品(肝脏、肾脏、脾脏、胸肌、腿肌、十二指肠、空肠和回肠)中没有CP4-EPSPS外源基因成分,同时综合分析表明,转基因豆粕对肉鸡的血清生化指标和器官发育没有显著差异,在试验中转基因豆粕没有在肉鸡体内残留,对生化指标和器官发育没有显著差异。     

生长猪对转基因豆粕外源DNA的代谢研究

研究了抗草甘膦转基因豆粕中的外源DNA在生长猪体内的代谢和沉积。选用去势公猪，于回肠末端安装简单T型瘘管，饲喂常规和转基因豆粕日粮，收集回肠食糜和粪便；另外进行生长猪饲喂试验42d，添加约30％常规或转基因豆粕，屠宰后采取组织样品，检测外源DNA残留。结果表明，在回肠食糜和粪便中检出源自抗草甘膦豆粕的外源cp4 epsps基因，但重现率均仅有6．7％，外源DNA的完整性在小肠后段很大程度上受到破坏；PCR检测下限为5Pg时，在生长猪肝脏、肌肉、脾脏和胸腺组织中无cp4 epsps基因残留，外源DNA已被代谢降解。     

大豆中转基因成分的定性PCR检测

实验利用CTAB法对大豆DNA进行提取,设计并合成引物对转基因大豆的内源基因Lectin、外源基因CaMV35S 启动子、NOS终止子和目的基因Cp4 EPSPS基因进行PCR检测,成功建立了大豆中转基因成分的检测方法。     

豆粕饲料中转基因成分的多重PCR快速检测

以大豆内源基因、35 S启动子基因、NOS终止子和35S-CTP4为检测对象,研究了对内源基因与外源基因的引物终浓度配比及退火温度对转基因豆粕多重PCR检测的影响.结果表明,建立的多重PCR方法能够准确的同时检测出豆粕中的内源基因和转基因成分.     

饲料中转基因成分的实时荧光PCR检测研究

本试验采用实时荧光PCR方法对饲料中的转基因成分进行了检测研究。对国产的14份猪饲料、3份水产饲料和3份荷兰进口代乳粉共20份样品进行了大豆、玉米、棉花、油菜和大米物种特异性基因和CaMV35S、NOS、FMV35S、NPTⅡ、PAT、BAR、GOX、CryⅠA(b)、CryⅠA(b)-CryⅠA(c)、rrsoy外源基因检测。在11份样品中检出转基因成分,其中8份样品中含有转基因大豆成分,2份样品中含有转基因大豆和转基因棉花成分,1份样品中含有转基因大豆、转基因油菜和转基因棉花成分。     

进口饲料中牛源性与转基因成分的检测

本文介绍了进口饲料中牛源性与转基因成分的PCR检测方法。该方法用异硫氰酸胍提取进口饲料样品DNA,以牛特异性引物扩增模板DNA,扩增产物为271bp,PCR产物经限制性酶切片段分析,与预期设想一致;方法灵敏度实验结果显示,饲料样品中含有0.1%的牛源性成分时仍能检出。同时,根据转基因农作物最常使用的外源基因设计引物,在进口饲料中检测出35S启动子和NOS中止子等转基因成分,具有快速、简便、准确等特点。     

饲用转基因豆粕对牛奶营养成分的影响及奶中外源基因转移检测

随机选择青岛市周边7个奶牛场,取精料和牛奶样品,检测精料的水分、粗蛋白、粗纤维、粗灰分、钙和总磷含量及牛奶干物质、粗蛋白和钙含量,采取荧光定量PCR法检测精料及牛奶中的豆粕外源基因。结果表明：7个牛场奶牛精料的常规营养成分水分、粗灰分、钙、总磷含量差异显著（P＆lt;0.05）,精料粗蛋白质、粗纤维含量、牛奶中的干物质、粗蛋白质和钙含量7个牛场之间差异不显著（P＆gt;0.05）。4个牛场精料中含转基因豆粕GTS40-3-2和2704品系外源基因,3个牛场精料中含转基因豆粕GTS40-3-2品系外源基因。牛奶中均未检测到相应外源基因。可见,饲用不同种类转基因豆粕精料对牛奶主要营养成分含量无显著影响,豆粕外源基因未向牛奶中转移。     

转基因稻谷外源蛋白在肉仔鸡体内的安全评价

为了研究肉仔鸡体内器官中是否存在转基因稻谷所表达的外源蛋白,为转基因水稻品系进一步的饲用或食用安全性研究奠定基础,试验将200只1日龄的AA肉仔鸡随机分成2组,试验组、对照组,分别饲喂转基因稻谷及其亲本配制的基础日粮,采用Western blotting方法对饲喂转基因稻谷及其亲本稻谷种子、饲料及其对应粪便,各组肉仔鸡的胸肌、腿肌、肝脏、肾脏、血液、腺胃、肌胃、十二指肠肠壁、空肠肠壁、回肠肠壁的转基因外源蛋白在肉仔鸡体内的表达情况进行了检测。结果表明:采用Western blotting技术未在肉仔鸡体内各器官中检测到转基因稻谷所转录表达的外源蛋白,转基因蛋白成分能够被机体完全消化,肉仔鸡粪便中未检测到所转录的外源蛋白,转基因蛋白不会通过粪便在自然界传播。     

青贮玉米籽实对蛋鸡生产性能、蛋品质和肠道内环境的影响

本试验旨在探讨青贮玉米籽实对蛋鸡生产性能、蛋品质和肠道内环境的影响.试验选用55周龄海兰褐蛋鸡200只,随机分成4组,每组5个重复,每个重复10只鸡.4组蛋鸡分别饲喂含0(对照组)、1％、2％和4％青贮玉米籽实的饲粮,试验期为7周.结果表明:与对照组相比,2％和4％青贮玉米籽实组产蛋率、平均日产蛋量、蛋壳颜色、哈夫单位均显著提高(P＜0.05),料蛋比显著降低(P＜0.05),盲肠和空肠中大肠杆菌数量显著降低(P＜0.05)、乳酸杆菌数量显著提高(P＜0.05);1％青贮玉米籽实组平均日产蛋量、哈夫单位、空肠与盲肠中乳酸杆菌数量显著提高(P＜0.05),空肠中大肠杆菌数量显著降低(P＜0.05);4％玉米籽实组空肠和盲肠pH显著降低(P＜0.05).以上结果提示,饲粮中添加适量青贮玉米籽实能够提高蛋鸡的生产性能和蛋品质,改善肠道内环境,综合各项指标可知,添加2％青贮玉米籽实效果最好.     

生物发酵饲料对蛋鸡生产性能和鸡蛋品质的影响

［目的］旨在研究生物发酵饲料对蛋鸡生产性能和蛋品质的影响。［方法］选取8 000只367日龄健康海兰褐蛋鸡,随机分成2组,每组4 000羽蛋鸡。第1组为对照组,饲喂基础日粮;第2组为试验组,饲喂在基础日粮中添加4%发酵棕榈粕的混合饲料,试验期为45 d。［结果］试验第45天,试验组较对照组平均日产蛋重提高6.62% （P〈0.05）,料蛋比降低4.56%（P〈0.05）;相比对照组,试验组蛋白高度、哈氏单位分别增加了9.56%（P〈0.05）和6.31%（P〈0.05）,差异显著;试验组蛋黄颜色等级高于对照组,差异极显著（P〈0.01）。［结论］在蛋鸡日粮中添加发酵棕榈粕可提高其平均日产蛋重、哈氏单位,增加蛋白高度,提升蛋黄颜色等级、蛋鸡产蛋性能和蛋品质量。     

Effect of dietary nonphytate phosphorus on laying performance and small intestinal epithelial phosphate transporter expression in Dwarf pink-shell laying hens

This study examined the effects of various levels of dietary nonphytate phosphorus on laying performance and the expression patterns of phosphorus metabolism related genes in Dwarf pink-shell Jaying hens. A total of 405 28-week-old Dwarf pink-shell laying hens were fed the same corn-soybean basal meals but containing 0.20%, 0.25%, 0.30%, 0.35% or 0.40% nonphytate phosphorus. The results showed that feed intake, egg production, and average egg weights were quadratically correlated with dietary nonphytate phosphorus content （P 〈 0.05）, and the highest egg production, feed intake and average egg weights were achieved when dietary nonphytate phosphorus was at 0.3% （P 〈 0.05）. mRNA expression of intestinal sodium phosphorus co-transporter linearly decreased when dietary nonphytate phosphorus increased, mRNA and protein expression of intestinal calbindin and vitamin D receptor correlated quadratically with dietary nonphytate phosphorus, and the highest expression was found when dietary available phosphorus was at 0.2,5% to 0.3%. In conclusion, the ideal phosphorus requirement for Dwarf pink-shell layer hens is estimated to be 0.3% in a corn-soybean diet. With this level of phosphorus supplementation, calbindin and vitamin D receptor reached their highest expression.     

Effect of enzyme preparation containing xylanase and beta-glucanase on performance of laying hens fed wheat/barley- or maize/soybean meal-based diets

1. A commercial enzyme preparation (Quatrazyme HP) containing xylanase and beta-glucanase was examined in two laying hen experiments with wheat/barley- or maize-based diets. The activities of other enzymes were measured also. Starch, cell wall contents and effects of Quatrazyme HP on in vitro viscosity of wheat, barley, maize and soybean meal were determined. 2. In the first experiment, 90 ISA Brown laying hens at 28 weeks of age were given a wheat/barley basal diet with or without 20mg of Quatrazyme HP, which provided 560 and 2,800 IU of xylanase and beta-glucanase/kg diet. In the second experiment, 66 ISA Brown laying hens at 45 weeks of age were given a maize/soybean meal basal diet with or without 20 mg of Quatrazyme HP/kg diet for 9 weeks. Egg production, egg weight, egg mass, feed conversion ratio and change in body weight were recorded as response criteria. 3. There was a significant improvement in feed conversion ratio with enzyme supplementation. Birds given an enzyme-supplemented diet gained 86 g while those fed on the unsupplemented diet lost 103 g of their body weight by the end of the experiment. 4. The enzyme preparation did not affect either egg production, egg weight or egg mass of birds fed on the maize/soybean meal diet. However, a significant improvement in feed conversion ratio was detected. Birds on either the supplemented or unsupplemented diet exhibited an increase in their body weight at the end of the experiment. 5. Addition of xylanase and beta-glucanase decreased in vitro viscosity of wheat, barley, maize and soybean meal. This effect was greater for wheat and barley than for maize and soybean meal. 6. It was concluded that the beneficial effect of using an enzyme preparation containing xylanase and beta-glucanase is not limited to wheat/barley-based diets but also occurs with maize/soybean meal-based diets.     

Effect of dietary defatted diatom biomass on egg production and quality of laying hens

Background： This study was to determine if feeding laying hens with defatted diatom microalgal biomass （DFA） from biofuel production affected their egg production and health status. Methods： Five replicates of 5 individually caged ISA Babcock White leghorn hens were fed 4 diets, including a corn-soybean meal control diet, a diet containing 7.5% DFA substituting for soybean meal, and diets containing 7.5% or 15% DFA substituting for corn and soybean meal. Body weights, feed intake, feed conversion ratio （FCR）, rate of egg production, egg size, egg mass, and several characteristics of eggs were determined at 4 and 8 wk. Venous blood was sampled at 4 and 8 wk for measurement of 5 biomarkers of health. Results： The15% DFA diet decreased （P 〈 0.05） feed intake, egg production, and plasma uric acid concentrations as compared with the control diet, but increased （P 〈 0.05） egg albumen weight and height compared with the 7.5% DFA diets. The two levels of DFA produced dose-dependent （P 〈 0.05） changes in three color measures of egg yolk, without affecting four hen plasma biochemical indicators of health. Conclusions： Feeding laying hens with 7.5% DFA in the corn-soybean meal diet for 8 wk had no adverse effect on their health, egg production, or egg quality, but 15% inclusion reduced feed intake, egg production, and efficiency of feed utilization.     

酶制剂、益生素对杂粕饲粮品质改善效果的研究

采用单因素完全随机设计,研究杂粕日粮中添加0.015%的酶制剂和0.15%的益生素与豆粕日粮饲喂蛋鸡的效果差异。结果表明,与豆粕相比,加酶或加益生素的杂粕日粮降低蛋鸡产蛋率、产蛋量、蛋重和蛋鸡采食量;通过添加益生素或酶制剂,杂粕日粮在饲料转化率上与豆粕日粮效果相当;与豆粕日粮相比,加酶或加益生素的杂粕日粮不影响蛋壳厚度和软破蛋率,也不影响能量和蛋白质代谢率,但显著降低排泄物中蛋白质含量。可见,一定程度上加酶或加益生素能可提高杂粕的利用率,但其实用效果与豆粕日粮还有一定差距。     

Effect of dietary defatted diatom biomass on egg production and quality of laying hens

Background

This study was to determine if feeding laying hens with defatted diatom microalgal biomass (DFA) from biofuel production affected their egg production and health status.

Methods

Five replicates of 5 individually caged ISA Babcock White leghorn hens were fed 4 diets, including a corn-soybean meal control diet, a diet containing 7.5% DFA substituting for soybean meal, and diets containing 7.5% or 15% DFA substituting for corn and soybean meal. Body weights, feed intake, feed conversion ratio (FCR), rate of egg production, egg size, egg mass, and several characteristics of eggs were determined at 4 and 8 wk. Venous blood was sampled at 4 and 8 wk for measurement of 5 biomarkers of health.

Results

The 15% DFA diet decreased (P <?0.05) feed intake, egg production, and plasma uric acid concentrations as compared with the control diet, but increased (P <?0.05) egg albumen weight and height compared with the 7.5% DFA diets. The two levels of DFA produced dose-dependent (P <?0.05) changes in three color measures of egg yolk, without affecting four hen plasma biochemical indicators of health.

Conclusions

Feeding laying hens with 7.5% DFA in the corn-soybean meal diet for 8 wk had no adverse effect on their health, egg production, or egg quality, but 15% inclusion reduced feed intake, egg production, and efficiency of feed utilization.     

大豆肽和乳酸菌素对蛋鸡生产性能、蛋品质及血液生化指标的影响

本试验旨在研究大豆肽和乳酸菌素对蛋鸡生产性能、蛋品质及血液生化指标的影响.选取336日龄健康罗曼商品蛋鸡576只,随机分成8组,每组3个重复,每个重复24只鸡.对照组饲喂基础饲粮,试验组分别饲喂在基础饲粮中添加0.075％、0.15％、0.3％大豆肽和0.1％、0.2％、0.4％乳酸菌素及0.15％大豆肽+0.2％乳酸菌素的试验饲粮.预试期1周,正试期4周.结果表明:1)饲粮中添加0.15％大豆肽或0.2％乳酸菌素均可显著提高蛋鸡产蛋率(P＜0.01),降低料蛋比(P＜0.05),显著提高哈夫单位(P＜0.05),对平均日采食量、蛋壳厚度、蛋壳强度无显著影响(P＞0.05).2)饲粮中添加0.15％大豆肽或0.2％乳酸菌素可以极显著或显著提高血浆谷草转氨酶(GOT) (P ＜0.01,P＜0.05)、谷丙转氨酶(GPT)活性(P＜0.01),降低血浆尿酸含量(P＜0.01),对血浆总蛋白、清蛋白、球蛋白含量无显著影响(P＞0.05).由此可见,饲粮中添加0.15％的大豆肽或0.2％的乳酸菌素均能提高蛋鸡生产性能和蛋品质,改善蛋白质代谢,并且0.15％大豆肽+0.2％乳酸菌素的联合应用优于单独添加大豆肽或乳酸菌素.     

产蛋鸡日粮内低芥酸、低硫甙菜籽粕营养价值的研究

本文分析了国产低芥酸、低硫甙菜籽粕营养及抗营养成分。选21周龄罗曼蛋鸡500只随机均分为5组 ,分别饲喂全豆粕和10 %、15 %、17 %、20 %双低菜籽粕日粮 ,试期35周。结果表明 :双低菜籽粕的蛋白质品质不及豆粕 ,硫甙及其分解产物仅为普通菜籽的1/3左右 ,植酸和单宁含量较高。在日粮中添加10 %、15 %双低菜粕对蛋鸡的生产性能、甲状腺、肝、肾及血清中TSH、T3、T4 和血浆中GPT、GOT的活性等均无异常影响(P>0.05)。添加17 %、20 %双低菜籽粕 ,蛋鸡产蛋量及料蛋比不佳(P<0.05) ,甲状腺、肝等器官有一定异常变化 ,但未见明显肿大(P>0.05)。血清T3、T4、TSH及血浆GOT、GPT无异常变化(P>0.05)。