基于ITS基因分析黑斑蛙裂头蚴种系发育关系

为了研究黑斑蛙裂头蚴种系发育关系,本研究利用聚合酶链式反应(PCR)对黑斑蛙裂头蚴湖南株核糖体DNA内转录间隔区(ITS)进行扩增与测序,应用ClustalX 2.0程序对序列进行比对,再利用Mega4.0程序进行NJ法绘制种系发育树,研究黑斑蛙裂头蚴与其他带科绦虫的种群遗传关系。结果显示,湖南省不同地区10个黑斑蛙裂头蚴分离株ITS序列基本一致,为1 362~1 382bp,各分离株之间ITS-1和ITS-2序列的同源性均高于95.4%和94.6%;与基因库中其他带科绦虫ITS-1和ITS-2序列的同源性均低于59.5%和56.9%。通过建立NJ系统发育树,湖南省黑斑蛙裂头蚴分离株与欧猬迭宫绦虫位于同一大分支,得到了很好的鉴定。黑斑蛙ITS序列在种内相对保守,而种间差异较大,因此ITS序列可以作为分子标记研究黑斑蛙裂头蚴种系遗传变异,从而为黑斑蛙裂头蚴的分子流行病学和群体遗传研究奠定基础。     

多头带绦虫湖南分离株核糖体18S基因系统进化分析

为阐明多头带绦虫湖南分离株核糖体小亚基(18S rRNA)基因序列的遗传变异情况,并用18S序列构建多头带绦虫与其他带科绦虫的种群遗传关系。利用PCR技术扩增多头带绦虫的18S基因,应用Clustal X1.81程序对序列进行比对,再利用Mega4.0程序进行NJ法绘制种系发育树。结果显示:所获得的18S基因序列长度一致,均为1 080 bp,基因序列存在一定的遗传差异。种系发育分析结果显示,所有的多头带绦虫分离株与已知多头带绦虫位于同一分支。多头带绦虫18S基因序列种内相对保守,种间差异较大,可以作为种间遗传变异研究的标记,从而为多头带绦虫的群体遗传研究奠定基础。     

基于ITS基因分析蛇源裂头蚴种系进化关系

为了研究蛇源裂头蚴种系发育关系,本研究对大王蛇裂头蚴湖南株的核糖体DNA内转录间隔区(ITS)进行扩增与测序,并对其基因序列进行比对后绘制种系进化树,分析蛇源裂头蚴与其它带科绦虫的种群遗传关系。结果显示,12个蛇源裂头蚴分离株ITS基因序列长度差异较小,为1 386 bp~1 404 bp;各分离株之间ITS-1和ITS-2差异分别为0.2%~1.3%和0.4%~2.2%,与Gen Bank中登录的其它带科绦虫相比,同源性均分别低于66.3%和52.1%。表明蛇源裂头蚴ITS基因序列在种内相对保守,种间差异较大,可以作为蛇源裂头蚴的分子遗传标记。分离株系统进化树显示,所有分离株与欧猬迭宫绦虫位于同一大分支,与其他带科绦虫得到了很好的鉴别。本研究为蛇源裂头蚴的鉴别诊断、分子流行病学调查等方面的研究奠定基础。     

扩展莫尼茨绦虫α-tubulin基因的克隆及序列分析

分离和鉴定扩展莫尼茨绦虫(Moniezia expansa)新基因,为进一步研究该基因的功能奠定基础。利用构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获取其全长cDNA序列;采用生物信息学等分析技术对该cDNA序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较及蛋白质二级结构的初步预测。获得了1个扩展莫尼茨绦虫新基因α-tubulin,全长1522bp,编码225个氨基酸,CDS预测属于Tubu-lin_FstZ超家族。编码蛋白的理论分子质量为25808.6KDa,等电点为9.15。获得了扩展莫尼茨绦虫α-tubulin的全长cDNA序列,为该基因功能的试验性鉴定工作奠定基础。     

豆状带绦虫线粒体rrnL和nad5基因的克隆及种系发育研究

本研究旨在阐明中国豆状带绦虫囊尾蚴河南分离株线粒体核糖体大亚基基因(rrnL)部分序列(prrnL)和烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶亚单位5基因(nad5)部分序列(pnad5)的遗传变异情况,并用prrnL和pnad5序列重构豆状带绦虫与其他带科绦虫的种系发育关系。采用聚合酶链式反应(PCR),扩增豆状带绦虫囊尾蚴分离株的线粒体prrnL和pnad5,将所测的序列用PAUP 4.0 Beta 10程序MP法绘制种系发育树,利用DNAStar 5.0中的MegAlign程序进行同源性分析。所获得的豆状带绦虫线粒体prrnL长度约为847 bp,pnad5为602 bp。所获rrnL序列与GenBank中豆状带绦虫相应序列的相似性为99.1%~100%,nad5为99.2%~100%;河南分离株之间rrnL序列的相似性为99.65%,nad5为99.5%;与GenBank中其他带科绦虫序列的相似度rrnL低于83.4%,nad5低于85.7%。种系发育分析结果表明,所有豆状带绦虫分离株和已知豆状带绦虫位于同一分支。由于豆状带绦虫分离株的线粒体rrnL和nad5基因序列种内很保守,种间差异较大,故可作为带科绦虫种间遗传变异研究的标记。     

扩展莫尼茨绦虫引发酶蛋白基因的克隆和cDNA序列分析

分离和鉴定扩展莫尼茨绦虫(Moniezia expansa)新基因,为进一步研究该基因的功能奠定基础。构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获取其全长cDNA序列;采用生物信息学等分析技术对该cDNA序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较及蛋白质二级结构的初步预测。获得了1个扩展莫尼茨绦虫新基因——引发酶蛋白,全长1269 bp,编码422个氨基酸,属于AE_Prim_S家族。编码蛋白的理论分子质量为47.1598 ku,等电点为4.83。获得了扩展莫尼茨绦虫反应结合蛋白的全长cDNA序列,为该基因功能的试验性鉴定工作奠定基础。     

蛇钩口线虫长沙分离株ITS基因的克隆及序列分析

为研究蛇钩口线虫长沙分离株的核糖体DNA (rDNA)内转录间隔区(ITS)序列的遗传变异情况,并利用ITS序列构建蛇钩口线虫与其它线虫的种群遗传关系,本研究利用PCR扩增蛇钩口线虫rDNA的ITS片段,并克隆至pGEM-T载体中进行序列测定及分析.结果显示,长沙市各分离株的ITS序列长度均为734 bp,与其它线虫的同源性均低于83.9％,而与十二指肠钩口线虫的同源性较高.本研究为蛇钩口线虫的分类鉴定以及进一步的分子流行病学调查和群体遗传研究奠定了基础.     

山羊扩展莫尼茨绦虫病的防治

山羊扩展莫尼茨绦虫病是由裸头科、莫尼茨属的扩展莫尼茨绦虫（Moniezia expansa）寄生于山羊小肠而引起的一种消化道寄生虫病。特别对羔羊危害严重，羔羊不但易感，感染后常会引起腹泻、贫血、消瘦、阻碍生长发育，严重可致死亡，造成很大的经济损失。现对一起山羊扩展莫尼茨绦虫病的防治报告如下。     

贝氏莫尼茨绦虫与扩展莫尼茨绦虫生殖器官形态学观察与比较

目的将染色制片的两种莫尼茨绦虫进行生殖器官形态学观察与比较。方法采用苏木素染色法，将采集的绵羊莫尼茨绦虫染色制片，将其分类为贝氏莫尼茨绦虫和扩展莫尼茨绦虫后，分别各选取10条进行形态结构测量与比较，同时对两种绦虫的节间腺进行了对比观察与测量。结果两种莫尼茨绦虫的卵巢在幼节的前段，距头节25～45mm处开始形成雏形，卵巢的退化在距头节1029-1217mm后的孕节前段消失；而睾丸的形成在距头节285-320mm后的幼节末段处，睾丸的退化则在距头节751mm后的成节后段及距头节1281mm后的孕节前段消失。结论观察结果说明，莫尼茨绦虫雌性生殖器官的发育早于雄性生殖器官的发育。对扩展莫尼茨绦虫节间腺的观察发现，有的绦虫的节间腺呈一排分布，也有的呈两排分布，还有个别绦虫无节间腺分布。     

绵羊莫尼茨绦虫病的诊断与治疗

绵羊莫尼茨绦虫病主要是由带科、莫尼茨属的扩展莫尼茨绦虫和贝氏莫尼茨绦虫寄生于绵羊小肠内而引起的一种绦虫病。本病主要危害羔羊,影响其生长发育,严重感染时,可导致死亡。2005年11月,某个体养羊户饲养的绵羊中,当年羊多数出现以精神不振,食欲下降,贫血消瘦,便秘和腹泻交替发生为主要症状的疾病,通过对羊群发病情况调查,临床症状观察和实验室检查,确诊为莫尼茨绦虫病。通过采用特药物治疗,使病情得了有效的控制。1基本情况明水县某个体养羊户饲养绵羊210只,其中当年羊90只,入冬以来,当年生的绵羊中大多数开始消瘦,贫血,腹泻与便秘交替,并…     

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 -