高原鼢鼠微卫星多态位点筛选及其通用性检测

跨种扩增是微卫星DNA标记获取的快捷有效途径之一。为了筛选出高原鼢鼠微卫星多态性位点,从已开发微卫星信息的鼹形鼠科和甘肃鼢鼠物种中进行了高原鼢鼠微卫星多态性位点及其通用性研究,结果表明:在87对微卫星位点中,只有54对进行了成功扩增,多态位点仅有9个,27个鼹形鼠科物种的微卫星在高原鼢鼠中并没有多态性,9个多态位点全来自于同属的甘肃鼢鼠。基于获得的9个多态位点,检测到高原鼢鼠的等位基因数在4~12,其平均观测杂合度(Ho)为0.544,期望杂合度(He)为0.671,多态信息含量为0.551,研究为高原鼢鼠的微卫星标记应用奠定了基础。     

利用混合模板筛选多态性微卫星的北极狐群体遗传学分析

在群体遗传学研究中,微卫星是重要的分子标记之一,由于其在近缘物种间具有一定的通用性,因此常被跨物种使用,但测试其多态性的过程非常繁琐。本研究采用混合模板扩增的方法,快速简便地评估微卫星位点的多态性,为开展群体遗传学分析获得有用的微卫星标记,以10只北极狐(Alopex lagopus)的DNA为材料,分别随机抽取2,3,5,8个和全部10个样品进行等量混合作为模板,用10个微卫星位点的引物进行扩增,以全自动测序仪进行电泳分型。结果表明:所有等位基因的电泳信号峰都清晰可辨。与单一模板扩增相比,混合模板扩增时,所有位点的复等位基因都能稳定地显示出来,不同批次试验之间的重现性达到100%。信号峰面积与等位基因频率呈线性正相关,可用以推断待测位点的基因频率分布情况。最高信号峰与最低信号峰的面积之比与混合样品数呈正相关,与多态信息含量呈负相关,提示混合样品的数量存在一定限度,到一定限度时低频等位基因无法检出。同时,通过这一关系,也可以推测位点的多态信息含量。本研究从10个位点中获得5个具有多态性的位点,3个可用于北极狐遗传分析。     

绵羊(ATAG)n四碱基微卫星位点的筛选及其在亲子鉴定中的应用

本试验旨在筛选绵羊高度多态性四碱基微卫星遗传标记,建立适用于绵羊亲子关系鉴定的实验体系。试验以小尾寒羊为主要研究对象,从已有的绵羊参考基因组序列出发,基于全基因组序列筛选法共筛选出53个（ATAG）_n四碱基重复微卫星位点,然后通过基因分型数据共筛选出30个扩增效果好、多态信息含量（PIC）丰富的四碱基重复微卫星位点;30个位点的基因分型结果表明,共扩增出253个等位基因,平均等位基因数为8.433,等位基因数均>5,多态信息含量在0.566~0.898,观测杂合度（Ho）范围在0.548~0.903,期望杂合度（He）范围在0.631~0.921,平均期望杂合度为0.776;哈代-温伯格平衡检验30个位点均处于遗传平衡状态。随后根据PCR的扩增效率从获得的30个多态性位点中筛选出22个微卫星位点用于亲权排除概率的计算,根据多态信息含量的大小由高到低依次增加位点数进行组合。结果表明,在两个亲本的基因型均未知的情况下,标记位点数为15个时,累积排除概率可达到99.99%,其中单个位点的第一非亲排除率（non-exclusion probability of the first parent,NE-1P）介于0.321~0.663之间。利用建立的亲子鉴定体系对16只具有系谱记录的小尾寒羊样本进行检测,结果共鉴定出4个具有高置信度的绵羊家系,鉴定结果与系谱记录完全一致。本试验为绵羊分子系谱的构建、亲子鉴定以及保障绵羊育种工作的正常开展奠定重要基础。     

黑龙江省完达山东部林区马鹿微卫星位点的筛选

利用黑尾鹿、白尾鹿等近缘物种的18对微卫星引物,对2个东北马鹿肌肉DNA样品和176个粪便DNA样品进行扩增,通过毛细管电泳法筛选适合东北马鹿的微卫星位点。结果表明,其中3个位点为高度多态性位点（PIC〉0.5）,1个为中度多态性位点（0.5〉PIC〉0.25）。共检测到26个等位基因片段,平均每个位点6.5个等位基因。多态信息含量0.393～0.827,个体识别率0.643～0.958,累积个体识别率为0.9999,累积非父排除率为0.9976。这4个多态性微卫星位点为东北马鹿的遗传研究提供了理论依据。     

应用高分辨溶解曲线(HRM)技术分析猪微卫星多态性

为了验证利用高分辨熔解曲线(high-resolution melting curve analysis,HRM)技术对猪微卫星标记进行基因分型的可行性和可靠性,使用非变性聚丙烯酰胺凝胶电泳(PAGE)方法对5个微卫星(SSRs)位点多态性进行检测,再使用HRM技术对这5个微卫星(SSRs)位点进行基因分型测定,最后对1个微卫星的所有基因型进行单克隆测序。结果表明:在样本测试中,使用HRM技术分辨出S0107和SW1953位点均有8种基因型,SW72位点有6种基因型,SW29位点有5种基因型,SW2155位点有4种基因型,其结果与利用非变性聚丙烯酰胺凝胶电泳方法检测的结果完全吻合,并且SW72位点的所有基因型克隆测序结果与HRM检测一致。这些结果提示,HRM分析技术可作为筛选猪微卫星标记多态性和对其进行基因分型的有效手段。     

南方大口鲶微卫星标记在怀头鲶群体中适用性研究

本研究分析了31对南方大口鲶（Silurus meriaionalis）微卫星标记引物在怀头鲶（Silurus soldatovi）群体中的适用性。结果表明,南方大口鲶微卫星标记引物在怀头群体中具有较高的适用性,共筛选出可以稳定扩增的微卫星标记15对,每个位点等位基因数为1～5个不等;不同重复拷贝类别的位点以富含AC重复单元的等位基因数较多,多态性比较丰富。借用已有的遗传标记资源,增加同目鱼类遗传连锁图谱上遗传标记的密度,将对今后开展怀头鲶种质资源保护及分子育种奠定坚实基础。     

中国地鼠微卫星DNA引物的设计及筛选

本研究旨在筛选中国地鼠微卫星位点,为中国地鼠遗传分析提供遗传标记。根据建立的中国地鼠微卫星富集文库,对筛选的含微卫星DNA序列的克隆应用引物设计软件Primer 3.0设计引物135对,选择合成11对理论上易出现影子带的引物,用20个中国地鼠个体对这些引物进行了评估。结果显示,11对引物均能扩增出谱带,这11条带的平均多态信息含量(PIC)为0.4195,平均观察杂合度(Ho)为0.3895,平均期望杂合度(He)为0.4565,每个位点的平均等位基因数为5.818。筛选出的微卫星位点均可用于中国地鼠种群遗传结构分析,这将为中国地鼠品种选育、种系评估提供更多的微卫星遗传标记信息。     

微卫星标记及其在家畜遗传多样性中的应用

<正>微卫星DNA标记是近年继RFLP之后发展起来的一种新的分子遗传标记技术。微卫星DNA标记具有数量大、分布广且均匀、多态信息含量高、检测快速方便等优点。在畜禽品种资源分类及遗传多态性评估和保存研究中,微卫星也是被广泛应用的遗传标记之一,通过对微卫星位点等位基因的突变模式研究来计算遗传距离,进行相关物种的系统进化研究以及利用微卫星位点重构系统发生树等。1微卫星标记的基本特征1.1微卫星DNA的结构、功能及类型     

山西瘦肉型猪SD-Ⅲ系微卫星标记SW489和SW2049的遗传多态性分析

微卫星标记(microsatellite)又称为短串联重复序列(si m-ple tandemrepeats,STRs),是均匀分布于真核生物基因组中的简单重复序列,由2~6个核苷酸的串联重复片段构成,由于重复单位的重复次数在个体间呈高度变异性并且数量丰富,因此微卫星标记的应用非常广泛〔1〕。微卫星位点通常通过PCR扩增,扩增产物通过电泳分析并根据大小分离等位基因进行检测。本试验研究的目的就是分析山西瘦肉型猪SD-Ⅲ系卫星标记SW489和SW2049的遗传多态性,从分子水平上了解山西瘦肉型猪SD-Ⅲ系,为猪的动物育种提供科学依据。1材料和方法1.1试验材料1.1.1试验猪…     

小麦族十一个物种的叶绿体psaC基因序列比较分析(简报)

植物叶绿体基因组的psaC基因有非常重要的功能,是植物光化学反应和光系统Ⅰ的组成成分。本研究对小麦族11个物种的psaC基因进行了扩增、测序和对比分析。结果表明,1)psaC基因编码区非常保守,存在3个SNP位点,但这3个突变位点都为同义突变,并没有引起氨基酸的改变。2)在psaC基因的间隔区,栽培大麦多了一个6 bp的‘CAAAAA’多核苷酸插入。在已经研究的植物中,该‘CAAAAA’多核苷酸插入是栽培大麦所特有的,表明该片段是在大麦物种分化以后插入的。该片段不但可以作为栽培大麦特异的标记序列,而且在大麦属物种的系统进化关系研究中有特殊的用途。3)发现的SNPs位点可用于相关物种的细胞质分子标记开发。     

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.     

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.     

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.     

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 -