从日本牛体分离的卵形巴贝斯虫新种(续完)

在双芽巴贝斯虫的病例中,红细胞和白细胞数的减少率分别为７４．６％和５７．６％,在所研究的４个种中,这个种出现了最明显的变化。相反,在大巴贝斯虫的病例中,平均寄生率和最高寄生红细胞数分别为２．１％和１．３×１０５个／ｍｍ３,在所研究的４个种中,这个种最...     

免耕播种紫花苜蓿更新割草地的效果

用草甘膦清除割草地中扁穗雀等杂草后免耕播种与常规耕耙后播各紫花苜蓿的效果及效益比较分析，认为前者比后者成本低，效果好，前者观测结果极其与后者的比例为，播种牧草上对重量８０．８０％，高３．６倍，紫花苜蓿优势度６９．８３％，高３．１倍，植被率９７．１０％，高１．４倍，地面处理及播种所需成本为２４０元／ｈｍ＾２低３．１倍。     

柔嫩艾美球虫（Eimeria tenella）体外细胞培养

试验比较了玻璃珠层析法，ＤＥＡＥ纤维素层析法，Ｇ３漏斗过滤法３种提纯柔嫩艾美球虫（Ｅｉｍｅｒｉａｔｅｎｅｌｌａ）子孢子的方法，柱高８ｃｍ，直径２００μｍ的玻璃株柱用ｐＨ８．０的Ｒｉｎｇｅｒ’ｓ缓冲液作洗脱提纯子孢子，回收率为６４．１％，子孢子洗脱高峰期集中，且易于无菌操作，柱高５ｃｍ的ＤＥＡＥ纤维素柱用ｐＨ８．０的甘氨酸缓冲液作洗脱液，提纯子孢子回收率为６３．２％，但严格无菌操作困难，Ｇ３漏斗法易     

黑龙江省犬肾膨结线虫的调查

以全身蠕虫学地在省内２０个县市共检查２１３只犬。结果在１２个县市的１８只犬体内检出３８条肾膨结线虫。感经８．４５％，感染强度１－６条，平均为２．１条，平均为２．１条，寄生１条的有１１只犬，占６１．１１％；寄生３条的３只犬，占１６．６３％；一５条的有２只犬，占１１．１１％；寄生２条和６条的１只犬分别为５．５６％。虫体的雄雌比例相当地１：２。寄生部位为右肾的２１条，腹腔的１３条，分别为５５．２６％和４     

添加民星超级料精饲喂育肥猪试验

民星超级料精系江西民星企业集团公司新近研制成功的，以提高畜禽对稻谷的利用率为主要目的饲料添加剂强化产品。经多点共２４８头育肥猪为期４５～８２天不等的试验表明，在基础日粮（含６０％以上谷粉）中添加１％民星超级料精，其增重提高１１．７％～１１７．６％，平均３６．６％，单位增重饲料费用节省２．６％～４８．４％，平均１８．４％。料肉比为２．４５～３．３４：１，平均３．２１：１。     

青海柴达木幼年双峰驼血细胞的显微观测

对青海省柴达木地区１－３月龄双峰驼血细胞的显微观察结果：（１）红细胞呈卵圆形或长卵圆形，大小为７．６７×４．１１（μｍ），体积略大于成年驼；（２）白细胞近似为圆形，大小和分类结果：嗜碱性白细胞７．０７×６．５４（μｍ）和０．２６±０．４９％，嗜酸性白细胞１１．４８×１０．３８（μｍ）和０．８４±０．９９％，幼稚核嗜中性白细胞，７．４７×６．８７（μｍ）和０．１６±０．４９％，杆状核嗜中性白细胞１０     

寄生于中国水牛Bubalus Vbubalis的巴贝斯虫一新种(梨形虫目:巴贝斯科)

经作者等深入研究，发现湖北省水牛巴贝斯虫病病原不是牛巴贝斯虫Ｂａｂｅｓｉａｂｏｖｉｓ与双芽巴贝斯虫Ｂ．ｂｉｇｅｍｉｎａ，而是一新种——东方巴贝斯虫Ｂａｂｅｓｉａｏｒｉｅｎｔａｌｉｓｓｐ．ｎｏｖ．。病牛红细胞中的虫体呈梨形（单梨形多于双梨形）、椭圆形、指环形、边虫形及杆状。以梨形虫体居多，病的初期其长度均小于红细胞半径，其平均大小为２．２×１．３μｍ，双梨形虫体尖端相连多排列成钝角或一字形，位于红细胞偏中央处。病的后期（出现虫血症后５～６ｄ）及体外培养７２ｈ，有少部份（１～２％）虫体直径大于红细胞半径。新种的传播者为镰形扇头蜱Ｒｈｉｐｉｃｅｐｈａｌｕｓｈａｅｍａｐｈｙｓａｌｏｉｄｅｓｈａｅｍａｐｈｙｓａｌｏｉｄｅｓ经卵传递（由第二代成蜱传给健康牛）。在成蜱的血淋巴中的虫样体（裂殖子）呈圆锥状、一端钝圆、尾部直而尖，不形成钩。新种对黄牛不能感染致病，只能使水牛致病。以１９９培养基加黄牛血清进行体外培养（微气静相培养技术）不能生长繁殖     

r—人工草地绵羊放牧系统优化生产模式研究：Ⅲ冬,春羔实验系统及优化系统的生

冬、春羔实验系统连续５年监测资料的平均结果表明，春羔系统的生产效率优于冬羔系统，前者每倾草的产出数量为５９９个畜产品单位；后者为５８４个ＡＰＵ；前者每公顷草地的利润为６７２．３元，后者为５６８．４０元；投资利润率分别为３５％和３０％。     

“871—3”超级浓缩料用于育肥猪的试验

“８７１—３”超级浓缩料系江西民星企业集团公司新近推出的猪用核心料，本试验在稻谷型日粮（含稻谷粉６０％以上）中添加４％“８７１—３”超级浓缩料，经多点共２０３头为期４５～８２天不等的试验表明，增重提高１７％～９１．６％，平均４０．０％，单位增重饲料费用节省２．６５％～３９．６％，平均１６．５％，料肉比为２．８６～３．６１：１，平均为３．６１：１。     

人工草地绵羊放牧系统优化生产模式研究Ⅲ冬、春羔实验系统及优化系统的生产效率分

冬、春羔实验系统连续５年（１９９０～１９９５）监测资料的平均结果表明，春羔系统（ＳＬＳ）的生产效率优于冬羔系统（ＷＬＳ），前者每公顷草地的产出数量为５９９个畜产品单位（ＡＰＵ）；后者为５８４个ＡＰＵ；前者每公顷草地的利润为６７２．３元，后者为５６８．４０元；投资利润率分别为３５％和３０％。基于系统监测资料的平均结果和已经达到的指标，对系统进行优化后，再将春羔系统与冬羔系统相比，则春羔系统的边际效益达５９０．９８元／ｈｍ２，利润为１１５９．３８元／ｈｍ２，比冬羔实验系统已获利润提高１．０４倍。     

Generally applicable methods to purify intracellular coccidia from cell cultures and to quantify purification efficacy using quantitative PCR

The objective of this study was to evaluate the utility of a simple, efficient, and rapid method for the isolation of Sarcocystis neurona merozoites and Besnoitia darlingi tachyzoites from cultured cells. The efficacy of this purification method was assessed by microscopy, SDS-PAGE, Western blotting, immuno-fluorescence, and three novel quantitative PCR assays. Culture medium containing host cell debris and parasites was eluted through PD-10 desalting columns. This purification method was compared to alternatives employing filtration through a cellulose filter pad or filter paper. The estimated recovery of S. neurona merozoites purified by the column method was 82% (+/-3.7) of the original merozoites with 97.5% purity. In contrast, estimated recovery of S. neurona merozoites purified by filter pad and filter paper was 40% and 30% with 76% and 83% purity, respectively. The same procedures were applied to purify B. darlingi tachyzoites from cultured cells. Of the original cultured B. darlingi tachyzoites, 94% (+/-2.5) were recovered from the PD-10 column with 96.5%, purity whereas percentage recovery of B. darlingi tachyzoites purified by filter pad and filter paper were 51% and 35% with 84% and 88% purity, respectively. All described methods maintained sterility so that purified parasites could be subsequently cultured in vitro. However, purification using a PD-10 column minimized parasite loss and the loss of viability as determined by the trypan blue dye exclusion assay, the rate of parasite production, and plaque forming efficiency in cell culture. Moreover, column-purified parasites improved the sensitivity of an immuno-fluorescent (IFA) analysis and real-time quantitative PCR assays targeted to parasite 18S ribosomal DNA and hsp70 genes. This technique appears generally applicable for purifying coccidia grown in cell cultures.     

In vitro antiprotozoal effects of artemisinin on Neospora caninum.

Neospora caninum is an intracellular apicomplexan parasite that infects a wide range of mammals and has been associated with abortion in cattle worldwide. Artemisinin is an effective antimalarial compound derived from a traditional Chinese herbal remedy, qinghao or Artemisia annua L. In the study reported, the cultured host cells (vero cells or mouse peritoneal macrophages) infected with N. caninum tachyzoites were incubated with alpha-MEM (minimal essential medium) 10%HS supplemented with various concentration or artemisinin (20, 10, 1, 0.1 and 0.01 microg/ml) to examine the efficacy of artemisinin against N. caninum tachyzoites intracellular multiplication. In long-term studies, at 20 or 10 microg/ml for 11 days, artemisinin reduced N. caninum and completely eliminated all microscopic foci of N. caninum. At 1 microg/ml for 14 days, artemisinin reduced N. caninum and completely achieved elimination of all microscopic foci of N. caninum. There was no apparent toxicity to host cells in long-term studies. In short-term studies, at > or = 0.1microg/ml, artemisinin reduced N. caninum tachyzoites intracellular multiplication, significantly (P < 0.05) and appeared to depend on the artemisinin concentrations. Pretreatment of host cells or N. caninum tachyzoites with artemisinin had no effect on N. caninum tachyzoites intracellular multiplication. These results demonstrate that artemisinin inhibited N. caninum tachyzoites intracellular multiplication.     

提高牦母牛繁殖性能的研究

采用不同剂量生殖激素处理3.5岁的青年牦母牛,结果发情率提高5.84和23.26个百分点,繁殖率分别提高34.69和41.21个百分点。采用不同剂量生殖激素处理和怀孕牦母牛围产期补饲的措施,促进产后成年牦母牛发情配种,结果发情率分别提高42.42,54.67和37.51个百分点,繁殖率提高43.93,48.48和20.10个百分点。生殖激素处理和围产期补饲对提高牦牛繁殖性能有较大的促进作用,高剂量生殖激素的促进效果较明显。     

Alterations of concentrations of calcium and arachidonic acid and agglutinations of microfilaments in host cells during Toxoplasma gondii invasion

Toxoplasma gondii (T. gondii) invasion of host cells is a complicated process of interaction between parasites and host cells. In the present study we investigated the alterations of free Ca(2+) concentration ([Ca(2+)](i)) and cytoskeletons in phagocytic and non-phagocytic host cells and arachidonic acid (AA) concentration in cells supernatant during T. gondii invasion. T. gondii invasion induced significant elevation of intracellular [Ca(2+)](i) in phagocytic cells (J774A.1) but not in non-phagocytic cells (L929). Pre-treatment of J774A.1 cells with Phospholipase C (PLC) inhibitor (U73122), or Ca(2+) chelators (EGTA, BAPTA/AM) did not block elevations of [Ca(2+)](i) but the elevations were lower and of shorter duration than that in untreated cells. Pre-treatment of tachyzoites with Phospholipases A (PLA) inhibitors (4-BPB and AACOCF3) resulted in a similar pattern of increasing of [Ca(2+)](i) as that in Ca(2+) chelators treated cells. Agglutinations of microfilaments were observed in J774A.1 cells but not in L929 cells. No changes of microtubules were observed in either cell. Treatment of cells with cytoskeleton inhibitors (colchicines, cytochalasin-D) resulted in reduced cell infection ratios. AA concentration in J774A.1 cells supernatant reached 8.44-fold of basal concentration after T. gondii infection and those in 4-BPB or AACOCF3 pre-treated cells reached 7.70-fold and 8.09-fold of basal concentration, respectively. However, elevation of AA concentrations induced by 4-BPB or AACOCF3 treated tachyzoites were 3.02-fold and 2.65-fold of basal AA concentration. AA concentration in L929 cells supernatant reached 5.02-fold of basal concentration after T. gondii infection and those in 4-BPB or AACOCF3 pre-treated cells reached 4.75-fold and 4.78-fold of basal concentration, respectively. However, elevation of AA concentrations induced by 4-BPB or AACOCF3 treated tachyzoites were 2.06-fold and 2.43-fold of basal AA concentration. Results indicated that elevations of [Ca(2+)](i) and AA induced by T. gondii invasion were from both host cells and parasites. T. gondii invasion activated host cell PLC and triggered the PLC-PKC signal pathway, which resulted in the flowing of extracellular Ca(2+) and the releasing of intracellular Ca(2+) pool. Elevated [Ca(2+)](i) induced reorganization of host cell microfilaments. The invasion also activated secretory PLA(2) (sPLA(2)) and cytosolic PLA(2) (cPLA(2)) of the parasite to release AA, which increased the permeability of cell membrane.     

淫羊藿－蜂胶合剂对鸡红细胞CR_1增强作用的实验研究

用不同浓度的淫羊藿－蜂胶合剂与鸡红细胞在体外进行孵育，然后测其红细胞补体Ｉ型受体（ＣＲ１）的活性，以判定淫羊霍－蜂胶合剂对红细胞免疫功能的增强作用。结果显示：经２０％、１０％和５％的淫羊藿－蜂胶合剂处理后的鸡红细胞ＣＲ１花环率（ＣＲ１－Ｒ）分别为：３８．９％、２２．０％和９．０％，面对照组鸡红细胞ＣＲ１－Ｒ是９．３％。此结果表明：淫羊藿－蜂胶合剂具有明显的增高红细胞ＣＲ１活性作用，并与剂量有关。     

淫羊藿－蜂胶合剂对鸡红细胞CR_1增强作用的实验研究

用不同浓度的淫羊藿－蜂胶合剂与鸡红细胞在体外进行孵育，然后测其红细胞补体Ｉ型受体（ＣＲ１）的活性，以判定淫羊霍－蜂胶合剂对红细胞免疫功能的增强作用。结果显示：经２０％、１０％和５％的淫羊藿－蜂胶合剂处理后的鸡红细胞ＣＲ１花环率（ＣＲ１－Ｒ）分别为：３８．９％、２２．０％和９．０％，面对照组鸡红细胞ＣＲ１－Ｒ是９．３％。此结果表明：淫羊藿－蜂胶合剂具有明显的增高红细胞ＣＲ１活性作用，并与剂量有关。     

露蕊乌头的种子萌发及幼苗出土对不同家系和亲代处理的响应

对青藏高原东部亚高寒草甸退化草地及河西走廊弃耕地上常见物种露蕊乌头(Aconitum gymnandrum)的种子萌发、幼苗出土及幼苗性状进行了研究,结果表明:1)亲代进行了去雄蕊和去花瓣处理延迟了子代幼苗的出土时间,同时,处理组的种子萌发率、幼苗返青率、叶面积及地上生物量都显著降低,但出苗率显著增加,此外,这些性状受亲代处理与家系间交互作用的影响显著,表明露蕊乌头幼苗生长特性有很强的遗传变异,具有较大的选择潜力;2)即将进入繁殖期的幼苗主要性状不受家系的影响,表明母本对露蕊乌头苗期性状的影响在不同阶段并不相同。     

生物肥部分替代化肥对花椰菜产量、品质、光合特性及肥料利用率的影响

高原夏季蔬菜生产面临着化肥过量使用导致的蔬菜品质下降、肥料利用率低和地下水污染等问题,通过研究生物肥部分替代化肥,探讨生物肥的增效作用、替代量和肥料利用率,为菜田合理施肥与提高蔬菜品质提供科学依据。采用随机区组试验,设不施肥(CK)、100%常规施肥(100CF)、80%常规施肥(80CF)、60%常规施肥(60CF)、100%常规施肥+生物肥(100CFB)、80%常规施肥+生物肥(80CFB)、60%常规施肥+生物肥(60CFB)7个处理,研究了化肥配施生物肥对花椰菜产量、品质、养分分配、肥料利用率及光合特性的影响。结果表明,与单施化肥相比,配施生物肥可显著提高花椰菜花球单重,同时提高了氮、磷、钾肥利用率及其吸收量;与100%常规施肥相比,化肥减施20%配施生物肥提高了花球产量,而化肥减施40%配施生物肥显著降低了花球产量,化肥减量配施生物肥显著降低了花球硝酸盐含量,显著提高了花球维生素C和可溶性糖含量;以80CFB处理氮肥利用率和功能叶吸氮量最高,100CF处理花球吸氮量最高,易造成硝酸盐的积累;配施生物肥增加了叶片净光合速率和气孔导度,减小了叶片胞间CO₂浓度。因此,适当减施化肥配施生物肥(80%常规施肥+生物肥)在不影响花椰菜产量的同时,能够显著改善花椰菜的品质、提高肥料利用率和光合效率。     

Monoclonal antibodies preventing invasion of Neospora caninum tachyzoites into host cells

Twelve monoclonal antibodies (MAbs) against Neospora caninum tachyzoites were produced to specify the antigens related to the invasion of tachyzoites into host cells. In the assay to evaluate the inhibition activity, all these MAbs prevented the cultured Vero cells from the invading by the tachyzoites. These MAbs recognized approximately a 73 kDa antigen in Western blot analysis. Immunofluorescence assay and immune electron microscopy revealed that this 73 kDa antigen is a part of the surface antigens of N. caninum tachyzoite, and that the tachyzoite antigen identified plays an important role for invasion of host cells.     

Repeatability and Reliability of Heart Rate Variability in Healthy,Adult Pony Mares

Heart rate variability (HRV) is an important noninvasive method to quantify stress by measuring sympathetic and parasympathetic activity of the autonomic nervous system. Few studies exist on the repeatability and reliability of HRV in equids. The objectives of this study were to (1) compare the effect of different correction factors (CF) available in HRV analysis software on HRV indicator values and (2) to determine the repeatability and reliability of HRV indicators in an unrestricted (pasture) and a restricted movement (equine examination stocks) environment. Data were recorded on five consecutive days from six adult pony mares. Short-term tachograms were compared with regard to software CF by graphical and statistical (Friedman's and Wilcoxon signed rank test) comparison. The results showed that the specific CF influences the HRV indicator values. The strong CF was able to balance the elimination of artefacts without removing the variability of RR intervals and was subsequently used to determine repeatability and reliability. HRV indicators showed good repeatability over the 5 days using Friedman test (pasture, P = .162–.898; examination stocks, P = .29–.865), indicating that there were no significant differences between HRV indicator values. The reliability, represented by intraclass correlation coefficient (ICC) and coefficient of variation (CV), was poor-to-good for pasture data (ICC = 0.44–0.79; CV = 10–68.10) and examination stocks data (ICC = 0.22–0.83; CV = 16.07–62.29). Measurements obtained from the examination stocks were less reliable than those from pasture. Using normalized low-frequency and normalized high-frequency components improved reliability. Free-movement environment-based HRV recordings could ensure better reliability but may require the use of a stronger CF.