根癌农杆菌介导的苜蓿遗传转化体系的建立

苜蓿基因型是限制遗传转化的关键因素之一。从13份苜蓿材料中筛选出一份具有高频再生潜力的基因型一公农1号,并以该品种为受体转化Na^＋／H^＋逆向转运蛋白基因,优化了遗传转化条件,建立了农杆菌介导的苜蓿遗传转化系统,获得了大量的转基因植株,分子检测证实目的基因已经整合到苜蓿基因组中。     

根蘖型苜蓿高频再生体系的建立

根蘖型苜蓿具有较强的耐牧性,是苜蓿育种重要的种质资源。建立高效的苜蓿再生体系有助于其遗传转化的研究。试验以公农4号苜蓿的子叶为材料,通过比较不同植物生长调节剂浓度配比对苜蓿愈伤组织诱导和分化的影响,建立了根蘖型苜蓿的高频再生体系。结果表明:最佳愈伤组织诱导培养基为:MS培养基+2 mg/L 2,4-二氯苯氧乙酸(2,4-D)+0.4 mg/L激动素(KT),子叶愈伤诱导率达100%;最佳分化培养基为:UM培养基+1.6 mg/L KT,愈伤组织分化率达50%;移栽小苗的成活率达90%以上。     

牛皮蝇HA抗原基因转化紫花苜蓿的研究

以“甘农1号”紫花苜蓿(Medicago sativa)7 d苗龄子叶和下胚轴为受体材料,建立了高效的苜蓿再生体系和遗传转化体系,筛选出MS+2,4 D 2.0 mg/L+6 BA 0.5 mg/L和MS+6 BA 0.5 mg/L+NAA 0.03 mg/L+GA3 2.0 mg/L为苜蓿子叶愈伤组织诱导和分化的适宜培养基;探讨了农杆菌浓度OD600约0.5、感染时间8~10 min、共培养时间2 d为适宜的转化条件。采用农杆菌介导法将Hyperdomin A（HA）基因导入紫花苜蓿,经PCR检测和Southern分子杂交分析,结果表明HA基因已经整合到苜蓿基因组中,可为牛皮蝇蛆病可食性疫苗的研究提供理论基础。     

九个苜蓿品种叶绿素荧光参数与叶水势日变化研究

在田间旱作条件下,对9个苜蓿品种的叶绿素荧光参数、叶片水势及其日变化进行了观测。结果表明:不同苜蓿品种的初始荧光(Fo)、可变荧光(Fv)、PSⅡ最大光能转化效率(Fv/Fm)、光合性能指数(PI)波动趋势呈单峰曲线,均在中午出现峰值,方差分析得出Fm、Fv、Fv/Fm、Fv/Fo以及PSⅡ活性中心数(RC/ABS)均表现出极显著差异(P<0.01),基于叶绿素荧光参数参试品种聚为两大类,第一类包括准格尔苜蓿、草原2号、敖汉苜蓿、中苜1号和龙牧801,叶绿素荧光特性较优,第二类包括肇东苜蓿、甘农3号、公农1号和龙牧803,叶绿素荧光特性较差;9个苜蓿品种水势日变化较大,其中准格尔苜蓿和敖汉苜蓿叶片水势较高且较为稳定,而公农1号和肇东苜蓿叶水势较低。准格尔苜蓿、草原2号、敖汉苜蓿、中苜1号和龙牧801对环境耐受性较好,肇东苜蓿、甘农3号、公农1号和龙牧803对环境耐受性较差。     

中国苜蓿繁育概述

回顾了建国以来国内苜蓿的育种发展状况,进行了苜蓿的种质评价与筛选,地方品种的系统化,育成了抗寒新品种草原1号、草原2号,抗病品种中兰一号,耐盐品种中苜一号,耐牧根蘖品种公农3号、中图3号、中图4号,高产的公农1号等.提出了在我国苜蓿育种方法上,杂交育种与基本育种方法在当代育种中将继续存在,生物技术在育种中的应用将加快育种进程;指出了我国苜蓿育种存在的问题是育种体系不健全,育种手段方法落后,抗性育种缓慢,对引入品种缺少理论与实践的鉴定、评价,国内育成品种少,高层知识力量薄弱等.     

口蹄疫病毒抗原决定簇融合基因转化苜蓿

摘要：通过农杆菌的介导,以携带O型和A型口蹄疫抗原决定簇融合基因O21 O14 A21 HBcAg的植物表达载体转化苜蓿(Medicago sativa)愈伤组织,并再生得到抗性植株。试验建立了苜蓿甘农1号愈伤组织再生系统;优化了农杆菌介导的苜蓿遗传转化系统;获得抗性再生植株4棵;GUS报告基因在转化的愈伤组织中得到瞬时表达;抗性植株PCR检测结果证实目的基因已经成功整合到苜蓿基因组中。     

公农1号等苜蓿品种抗寒高产性能比较--筛选东北寒冷地区适宜品种

本文就公农一号等苜蓿品种的抗寒和高产性能进行了比较,认为在东北适宜的抗寒高产苜蓿品种有:公农一号、公农二号、肇东、龙牧801和龙牧803等5个品种,可以在生产上大面积推广应用。     

中国苜蓿繁育概述

回顾了建国以来国内苜蓿的育种发展状况,进行了苜蓿的种质评价与筛选,地方品种的系统化,育成了抗寒新品种草原1号、草原2号,抗病品种中兰一号,耐盐品种中苜一号,耐牧根蘖品种公农3号、中图3号、中图4号,高产的公农1号等。提出了在我国苜蓿育种方法上,杂交育种与基本育种方法在当代育种中将继续存在,生物技术在育种中的应用将加快育种进程;指出了我国苜蓿育种存在的问题是育种体系不健全,育种手段方法落后,抗性育种缓慢,对引入品种缺少理论与实践的鉴定、评价,国内育成品种少,高层知识力量薄弱等。     

赤霉素浓度对苜蓿种子萌发的影响

以公农1号紫花苜蓿和呼伦贝尔黄花苜蓿2种苜蓿品种为材料,采用培养皿纸上发芽法,研究不同浓度赤霉素处理（以蒸馏水处理作为对照）对苜蓿种子萌发和幼苗生长的影响。结果表明：公农1号紫花苜蓿种子发芽势、发芽率、发芽指数、发芽速度以及根长随着赤霉素浓度的增加表现出先增加后降低的趋势,赤霉素浓度为50 mg/L时效果最好;呼伦贝尔黄花苜蓿种子发芽势、发芽率、发芽指数、发芽速度以及根长随着赤霉素浓度的增加表现出逐渐增加的趋势,赤霉素浓度为200 mg/L时效果最好。     

根癌农杆菌介导的紫花苜蓿遗传转化体系的建立与优化

以“中苜1号”紫花苜蓿7日龄无菌苗子叶和再生无菌苗的叶片为材料，建立了适用于农杆菌介导的转基因组织培养体系，并对MsNHX1基因进行转化。转化优化条件为：“中苜1号”紫花苜蓿7日龄无菌苗子叶、再生无菌苗叶片，用农杆菌菌液（A600=0.6）侵染6min，然后在培养基上铺一层灭菌滤纸培养7d后清洗，建立了苜蓿快速有效的遗传转化体系。     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

Prevalence of gastric lesions in racing Alaskan sled dogs

Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

Evaluation of biohydrogenation rate of canola vs. soya bean seeds as unsaturated fatty acids sources for ruminants in situ

An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.     

Serum biochemical values and mineral element contents of tissues in yaks

Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein     

Avian brood parasitism — a growing research area in behavioral ecology (part Ⅱ)

正We are pleased to publish the second special issue on avian brood parasitism and to be responsible guest editors for the two special issues of Chinese Birds (Vol. 3, No. 4, 2012 and Vol. 4, No. 1, 2013), entitled "Avian Brood Parasitism - A Growing Research Area in Behavioral Ecology". The first issue was published in December 2012. The goal of the two special issues is to publish accumulated knowledge and some of the recent developments in     

Internal Parasites and Haematological Values in Cattle Slaughtered in Buea Subdivision of Cameroon

Tropical Animal Health and Production -