基于高光谱信息融合和相关向量机的种蛋无损检测

为了尽可能早的检测出无精蛋和受精蛋,该文提出采用透射高光谱成像技术,融合图像和光谱信息,对其受精信息进行检测。利用高光谱图像系统采集孵化前种蛋在400~1 000 nm的高光谱图像,提取图像特征(长短轴之比、伸长度、圆度、蛋黄面积与整蛋面积之比);筛选出400~760 nm的波段,通过Normalize预处理结合相关系数法提取155个光谱特征变量;运用主成分分析法对图像和光谱的融合信息进行降维,采用相关向量机(relevance vector machine,RVM)分别建立基于图像、光谱和图像-光谱融合信息的受精蛋和无精蛋分类判别模型,并与支持向量机(support vector machine,SVM)模型进行比较,RVM模型检测正确率分别为90%,91%,96%;测试集检测时间分别为0.6619,1.0821,0.5016 s。SVM模型检测正确率分别为84%,90%,93%;测试集检测时间分别为5.9386,5.9886,5.6672 s。结果表明,基于图像-光谱融合所建立的模型优于单一信息的模型,在分类精度上,采用RVM分类精度高于SVM的分类精度;在分类时间上,RVM的分类时间比SVM短,因此,利用高光谱融合信息和相关向量机可以提高种蛋检测精度,研究结果为孵前无精蛋和受精蛋的在线实时检测提供参考。     

基于非损伤微测技术监测贮期鸡蛋氧呼吸规律

鸡蛋作为一个生命体,呼吸作用是其产后重要的生理活动,也是影响鸡蛋贮运效果的重要因素,研究鸡蛋氧呼吸规律对其贮藏保鲜意义重大。该文应用非损伤微测技术,搭建特有的微测试验平台,实时监测鸡蛋与外界微环境的O2交换速率以及交换强度。通过预试验选定探针与鸡蛋表面的最佳距离为50μm,选定鸡蛋最佳测试位置为钝端。试验选取一天中的4个时间点(6:00,12:00,18:00,24:00)来测试鸡蛋O2呼吸,借以研究鸡蛋在一天不同时间点的氧呼吸强弱;另外在每天同一时间点测试鸡蛋O2流速,连续测一个月,来研究鸡蛋贮藏期间的氧呼吸变化规律。试验发现:一天中鸡蛋的氧呼吸存在凌晨高下午低的规律性,无论是受精蛋还是非受精蛋,在每天的不同时间点呼吸强度有差异,受精蛋比非受精蛋的生命活动及新陈代谢变化更强烈,昼夜呼吸差异显著;一个月中,鸡蛋在贮期会产生2次呼吸峰值,分别出现在质变的转折点,并且第2次的峰值明显高于第1次的峰值。试验结果表明鸡蛋存在自身特有呼吸规律,生命代谢活动强时其对应氧呼吸活动也强,反之则弱;刚产下的鸡蛋的呼吸较为活跃。该研究可为如何优化贮藏条件以及提高种蛋的孵化率提供参考。     

低剂量γ辐射对鸡产蛋性能的影响

1985—1990年间,用~(60)Coγ源辐照滨白鸡种蛋,结果表明,辐照促进受精蛋胚胎发育,孵化率提高6.86％,健雏率提高5.52％,雏鸡增重5—6％,提高辐照当代母鸡产蛋率8.03％,辐照二代(商品代)母鸡开产日龄提前6—7周,拉长产蛋期,72周龄产蛋量提高10.48％。     

不同孵化温度对雏番鸭第二性比及孵化率的影响研究

试验研究孵化前期(1~10d)7个不同温度(36.3~39.3℃)对雏鸭出壳时第二性比(♂∶♀)和受精蛋孵化率的影响结果表明,不同孵化温度出壳的雏番鸭公母性别存在明显差异(P<0.01),且公雏率随温度升高而升高,母雏率随温度升高而下降;孵化前期温度为36.3~37.3℃时出壳雏番鸭公少于母,第二性比介于0.24~0.95;而孵化前期温度为37.8~39.3℃时出壳雏番鸭公多于母,第二性比为1.39~5.38;且只有在温度37.3℃和37.8℃时第二性比与遗传性比(1∶1)无差异(P>0.05),而其他温度时第二性比均表现明显差异(P<0.05或P<0.01)。雏番鸭第二性比与孵化前期温度间相关指数R2为0.99,预测误差率为-34.88%~ 14.58%间,说明调控孵化温度可在一定程度上控制雏番鸭第二性比,孵化温度对受精蛋孵化率有明显影响,36.3~37.8℃内孵化率随温度的升高而升高,在37.8~39.3℃内则随温度的升高而降低。     

水牛和黄牛种间体外受精的研究

将从屠宰母牛卵巢上采集的水牛和黄牛的卵母细胞经 2 0～ 2 4 h体外成熟培养后 ,分别用么拉水牛或奶牛精液进行种间和种内体外受精。受精卵在体外条件下继续培养到囊胚和孵化囊胚阶段。结果表明 ,用黄牛精子受精水牛卵母细胞 (黄×水 )或用水牛精子受精黄牛卵母细胞 (水×黄 )的种间体外受精均有较高的卵裂率 (分别为 4 6.5%和 68.4 % )。但其体外囊胚发育率分别只有 3.5%和 8.7% ,显著低于种内受精组(水×水组为 1 9.9% ,黄×黄组为 2 8.0 % ,P<0 .0 5)。在受精后 2 0 h对受精卵进行固定、染色 ,发现种间受精的黄×水和水×黄组 ,分别有 36.8%和 75.0 %的受精卵形成有 1个以上原核 ,且这些受精激活卵中大部分含 2个原核以上 (分别为 80 .9%和 80 .0 % ) ,说明种间受精的精子已进入到卵母细胞内受精并形成了雄性原核。     

体细胞因子对牛卵母细胞体外受精的影响

本研究系统探讨了体细胞因子对牛卵母细胞体外受精的影响。在受精液中添加５％或１０％的牛卵泡液（ＢＦＦ）明显降低牛卵母细胞受精后的分裂率（３８．３％和４０．４％比对照８７．５％，Ｐ＜０．００１）和囊胚发育率（１７．４％和８．３％比对照４０．４％，Ｐ＜０．００１）。然而，在受精液中加入５０％的内膜细胞受精条件液明显提高牛卵母细胞受精后的囊胚发育率（４３．１％比３４．８％，Ｐ＜０．０５），虽然该处理对牛卵母细胞受精后的分裂率无明显影响。在体外受精系统中加入颗粒细胞的受精条件液，新鲜的输卵管细胞，输卵管细胞的单层细胞或输卵管细胞的受精条件液对牛卵母细胞的分裂率及囊胚发育率均无明显影响。相反，新鲜输卵管细胞组获得较低的分裂率（８１．７％）和囊胚发育率（３２．２％）。这些结果表明：ＢＦＦ对牛卵母细胞的体外受精具抑制作用，而内膜细胞的条件液则具促进效应。ＢＦＦ的这种受精抑制因子可能来自血清而不是卵泡细胞。     

60Coγ射线对受精鸡蛋及当代鸡生长变化的影响

利用＾６０Ｃｏγ射线对受精鸡蛋进行了不同剂量的刺激，结果表明，不同品种受精鸡蛋的出雏率，孵化率在３００ｒａｄ达到最大值，之后随剂量的加大而降低，受精鸡蛋经低剂量刺激有利于雄性鸡的生长，对雌性鸡生长有抑制作用，较高剂量能促使当代鸡提早开产下蛋，这些变异能否遗传后代，有待进一步研究。     

巷道式孵化的工艺技术探讨

巷道式孵化以其独特的气流方式区别于厢式孵化。巷道式孵化器的气流把含氧量最多的新鲜空气带给老的胚胎，把各阶段老胚胎释放的热量又供给需热的各阶段胚胎的发育，并始终保持着理想的设计温度。因而，所需功率相对较小，且孵化容量大，可以节约大量的能源，具有良好的经济性。讨论了进行巷道式孵化的孵化厅布局及孵化操作技术。中国农业大学正大肉鸡发展中心采用巷道式孵化器对７批艾维茵父母代种鸡的３２０万枚种蛋进行孵化，获得了优异的孵化成绩，受精蛋孵化率平均为９４．７５％，入孵蛋孵化率平均为８５．６７％，健雏率平均为９８．２８％。     

巷道式孵化的工艺技术探讨

巷道式孵化以其独特的气流方式区别于厢式孵化。巷道式孵化器的气流把含氧量最多的新鲜空气带给老的胚胎，把各阶段老胚胎释放的热量又供给需热的各阶段胚胎的发育，并始终保持着理想的设计温度。因而，所需功率相对较小，且孵化容量大，可以节约大量的能源，具有良好的经济性。讨论了进行巷道式孵化的孵化厅布局及孵化操作技术。中国农业大学正大肉鸡发展中心采用巷道式孵化器对７批艾维茵父母代种鸡的３２０万枚种蛋进行孵化，获得了优异的孵化成绩，受精蛋孵化率平均为９４．７５％，入孵蛋孵化率平均为８５．６７％，健雏率平均为９８．２８％。     

抗精子sp18单克隆抗体对小鼠体外受精胚胎发育的抑制作用

哺乳类受精时，精子的多种膜蛋白、顶体蛋白和胞质蛋白参与了受精过程，因此，研究人员的目光都集中在这些功能蛋白在睾丸中的生成、附睾中的水解和扩散、顶体反应中的作用和扩散以及它们在受精过程中的主要功能等。然而，对于精子蛋白在受精后的命运这个基本问题的研究，几乎是个空白领域。可以肯定的是，精子的顶体区膜蛋白和顶体蛋白在受精过程中被释放；胞质蛋白和顶体内膜蛋白通过胞吞作用进入卵子内部，其中，胞质中的钙振荡蛋白可以激活卵子，是新生命的发动者；顶体后区和尾部膜蛋白伴随着受精过程掺入卵子质膜，演化成受精卵的质膜蛋白，对于这类蛋白的最终结局和潜在的功能，我们的认识还很有限，英国生物学家发现，大鼠精子尾部的2D6膜蛋白（23 kD）掺入受精卵质膜后，随着胚胎发育过程而逐渐弱化，至2-细胞期时，胚胎表面的2D6完全消失，因此无法研究该抗原在胚胎发育中是否具有生理功能，这方面的研究工作处于停滞状态。     

Antimicrobial potential of egg yolk ovoinhibitor, a multidomain Kazal-like inhibitor of chicken egg

Chicken egg ovoinhibitor is a multidomain Kazal-type serine protease inhibitor with unknown function. Comparison of expression between different tissues indicated that ovoinhibitor is highly expressed in the magnum and liver followed by the uterus, which secrete egg white, egg yolk, and eggshell precursors, respectively. The results also revealed that ovoinhibitor expression is increased in the liver during sexual maturation followed by a subsequent decrease in mature hens. Ovoinhibitor was purified from the egg yolk plasma from nonfertilized eggs using two consecutive affinity chromatographies and gel filtration. Purified egg yolk ovoinhibitor was shown to inhibit trypsin and subtilisin. It was shown that purified egg yolk ovoinhibitor exhibited antimicrobial activities against Bacillus thuringiensis . The results suggest that this anti-protease plays a significant role in antibacterial egg defense against Bacillus spp., preventing contamination of table eggs (nonfertilized eggs) and protecting the chick embryo (fertilized eggs).     

模拟微重力对鹌鹑胚胎原始生殖细胞及内脏器官发育的影响

本实验利用回转器旋转模拟微重力的生物效应，以鹌鹑胚胎作为实验材料，研究了鹌鹑胚胎生殖腺内原始生殖细胞的数目及内脏器官的组织学变化。结果表明，回转器旋转可使早期胚胎血液和生殖腺内的原始生殖细胞数减少;生殖腺、心脏、肝脏的组织学结构发生不同程度的损伤，但对肺脏，肾脏没有太大影响。     

利用shRNA真核表达载体干涉雌性鸡胚性腺中芳香化酶基因(CYP19A1)的表达

本研究通过把构建的短发夹结构RNA(shRNA)真核表达载体导入鸡胚,检测鸡胚性腺分化期质粒载体在鸡胚体内代谢情况及雌性鸡胚性腺中芳香化酶基因(CYP19A1)mRNA表达效率,进而探讨利用该方法在鸡胚体内进行特定基因干涉的可行性.实验针对CYP19A1基因构建了4条特异性表达载体,一条非特异性表达载体.每个实验组选取45个新鲜种蛋作为实验材料进行胚盘下腔注射,并设立空白对照组.鸡胚发育12d时,检测各处理组鸡胚肝脏组织中质粒存在情况,并取其左侧性腺组织进行目标基因的荧光定量分析.研究结果表明,导入组在12日胚龄时所有鸡胚基因组中均可检测到绿色荧光蛋白基因(EGFP);荧光定量结果显示,导入特异性表达载体cyp-580、cyp-1083和cyp-1295后,对应雌性鸡胚性腺CYP19A1 mRNA表达效率显著低于空白对照组,干涉效率分别为:73％、52％和85％;特异性表达载体cyp-1403组CYP19A1mRNA表达效率与空白对照组相比有所降低但无显著性差异.本实验为诱导鸡胚性反转提供了新方法并建立了鸡胚发育期特定基因体内干涉新平台.     

Residue analysis of tetracyclines and their metabolites in eggs and in the environment by HPLC coupled with a microbiological assay and tandem mass spectrometry

Tetracyclines are widely used in farm animals. This can cause drug residues in products of animal origin and, after excretion of these substances, in animal slurry and in soil fertilized with that slurry. In this paper, we present a method based on a microbiological assay coupled with HPLC for the detection of oxytetracycline, tetracycline, and chlortetracycline in eggs. After a simple liquid extraction of the samples and HPLC separation, fractions were collected on microtiter plates, and the tetracyclines were analyzed using the Staphylococcus aureus assay. This method was able to identify residues of tetracyclines in eggs at a level set by regulatory agencies (i.e., 200 microg/kg). In addition, it was shown that the described microbiological method can be used as a screening assay for the detection of tetracyclines and possible biologically active metabolites in animal slurry and soil samples. Employing the same extraction procedure, it was demonstrated that LC-MS-MS allowed the quantification of 20-400 microg/kg in eggs with recoveries ranging from 71 to 109% and RSDs of 3-15%.     

用冻干精子生产ICSI小鼠

前期研究中，我们采用小鼠卵母细胞胞浆内精子注射（ICSI）技术，成功地用新鲜精子获得了生理健康的ICSI小鼠。在此基础上，本文结合细胞冻干技术，进行了冻干精子ICSI生产试管小鼠的尝试。B6D2F1成年公鼠的精子在Tris-HCl-EDTA（pH8.2）溶液中冻干4 h后解冻，将其精子头注入到KM小鼠成熟卵母细胞的胞质中。6 h后，83.0%的卵子受精。用CZB溶液体外培养发现，冻干精子生产的胚胎，体外发育至2-C (92.0% vs 99.5%)、4-C（52.7% vs 97.2%）、桑椹胚（36.6% vs 86.3%）和囊胚的比例（21.4% vs 68.7%）极显著地（p<0.01）低于新鲜精子生产的ICSI胚胎。移植2-C期冻干精子ICSI胚胎检测体内发育，结果发现，D10的着床率为63.0%（17/27）；胎鼠的出生比例为21.7%（13/60），这些ICSI小鼠成年后的生理和生殖能力正常。试验结果说明，精子冻干后，仍能生产ICSI动物，冻干技术可以用于哺乳动物精子的保存。     

Identification and characterization of the precursor of chicken matrix metalloprotease 2 (pro-MMP-2) in hen egg

Using zymography and mass spectrometry, we identified for the first time the precursor of chicken matrix metalloprotease 2 (pro-MMP-2) as a complex with TIMP-2 (tissue inhibitor of metalloproteinases) in egg white and yolk. Real-time polymerase chain reaction confirmed that MMP-2 and its inhibitors TIMP-2 and TIMP-3 were expressed all along the oviduct and in the liver of laying hens. We also demonstrated that the processing of pro-MMP-2 into mature MMP-2 by serine proteases does not occur in vivo, although purified pro-MMP-2 undergoes proteolytic maturation by these proteases in vitro. Moreover, the relative pro-MMP-2 activity assessed by gelatin zymography was shown to decrease in egg white during the storage of unfertilized or fertilized eggs. However, the mature form of 62 kDa MMP-2 could not be detected. The fact that MMP-2 is found as a proform in fresh eggs suggests that the activity of this metalloprotease is regulated under specific conditions during embryonic development.     

Interaction of iron chelate and nitrogen fixation in peanuts grown on calcareous soil

The interaction of iron (Fe) nutrition and nitrogen (N₂) fixation was examined in peanuts grown in the field for two growing seasons. The treatments were: a) inoculated with Rhizobium, not fertilized with N, b) uninoculated, fertilized with N, and c) uninoculated not fertilized with N. These treatments were tested with or without Fe chelate application. Inoculated peanuts produced up to 42% higher N yield than the uninoculated, non‐fertilized plants. Moderately chlorotic peanuts fertilized with Fe did not increase significantly their yield but had bigger nodules than peanuts not fertilized with Fe. There were no interactions between Fe and N treatments, indicating that both nutrients were important for growth and for N₂ fixation. Remedy of Fe chlorosis on calcareous soils with FeEDDHA will not reduce N₂ fixation.     

Melamine/urea and oxamide fertilization of Kentucky bluegrass

Abstract

The commercial lawn care industry represents a large market for N sources. A formulated melamine (2,4,6‐triamino‐s‐triazine) plus urea combination (MLU) (45% melamine by weight) and oxamide were evaluated for use by the lawn care industry by comparing turfgrass response from these fertilizers to that from urea, sulfur coated urea (SCU), ureaformaldehyde (Nitroform), and a non‐fertilized check. Fertilizers were applied four times per year to field plots of Kentucky bluegrass (Poa pratensis L.) growing on a Flanagan silt loam (fine, montmorillonitic, mesic Aquic Argiudoll) at a rate of 49 kg N ha^‐1 per application. Color ratings and clipping weights were determined weekly during the growing season. Trends in color ratings paralleled those in clipping weights. During the first year of the study, MLU treated turf received significantly higher color ratings than nonfertilized turf on only 38% of the rating dates; this increased to 76% in the third year of the study. Color ratings for MLU fertilized turf compared more closely to ratings for Nitroform fertilized turf than for ratings for urea or SCU fertilized turf. The turf fertilized with oxamide received higher color ratings than Nitroform or urea fertilized turf and compared favorably with SCU fertilized turf. Programs utilizing MLU would require either high initial application rates or supplemental applications of another N source to provide acceptable results. Oxamide appeared suitable for use by the lawn care industry.