籽鹅卵巢组织中铁蛋白重链基因和8个新ESTs的定量研究

采用实时定量PCR技术定量检测产蛋前期和产蛋期籽鹅卵巢组织中铁蛋白重链基因(FTH)和8个新ESTs(ODEUG01~ODEUG08)的mRNA表达水平。结果表明:产蛋期籽鹅卵巢组织中FTH和8个新ESTs的相对表达量分别比产蛋前期卵巢组织高13.25、7.68、11.82、14.23、8.25、15.14、9.78、6.48、14.21倍。本研究进一步证实,FTH和8个新ESTs在产蛋期籽鹅卵巢组织中高效表达,FTH和8个新ESTs可能参与籽鹅卵巢功能的调节,并影响籽鹅的产蛋性能。     

黑羽番鸭PRL、PRLR基因序列分析及表达规律研究

催乳素(Prolactin,PRL)由动物垂体前叶分泌,与其受体(Prolactin receptor,PRLR)相结合才能作用于靶细胞。为了解PRL与PRLR基因在黑羽番鸭不同繁殖阶段各组织内的时空表达规律,利用RACE扩增方法获得了PRL与PRLR基因的CDS区序列,同时应用荧光定量与免疫组化相结合的方法,对产蛋前期、产蛋期、就巢期的黑羽番鸭垂体、卵巢、子宫中PRL与PRLR基因的表达量进行了比较分析。结果显示:PRL与PRLR基因序列在多种禽类中高度同源;PRL基因在垂体、卵巢、子宫中m RNA的表达具有明显的空间特异性,PRL基因在垂体中表达量最高,极显著高于卵巢和子宫中的表达量(P0.01),这种趋势贯穿产蛋前期、产蛋期和就巢期。PRLR基因在不同时期各组织内m RNA的表达具有明显的时空特异性。PRLR基因的表达量在垂体、卵巢中的表达量普遍较低,子宫中的表达量最高。PRLR基因在不同时期垂体和卵巢中的表达量没有明显的差异(P0.05),而子宫中的表达量在就巢期显著高于产蛋前期与产蛋期(P0.05)。     

鹅不同繁殖时期GnRH和GnIH基因表达和激素浓度变化分析

旨在研究GnRH和GnIH基因和激素在鹅繁殖过程中的重要作用,本研究以产蛋性能差异显著的四川白鹅和溆浦鹅为试验材料,利用Real-time RCR方法检测产蛋前期、产蛋期和休产期两种鹅的下丘脑、垂体和卵巢组织中GnRH和GnIH mRNA的表达量;利用放射性免疫和酶联免疫吸附测定法测定血清中的GnRH和GnIH激素变化情况。结果表明:GnRH和GnIH基因在四川白鹅和溆浦鹅的下丘脑、垂体和卵巢组织中均有表达;产蛋前期和产蛋高峰期的下丘脑和卵巢组织中,四川白鹅的GnRH mRNA表达量显著(P0.05)或极显著(P0.01)高于溆浦鹅,在产蛋高峰期和休产期四川白鹅GnRH激素浓度也极显著(P0.01)或显著高于(P0.05)溆浦鹅,在高峰期和休产期四川白鹅GnRH血清浓度分别为51.13和51.10pg·mL-1,溆浦鹅分别为49.52和49.94pg·mL-1;GnIH在两种鹅的变化比较一致,仅在产蛋高峰期,四川白鹅下丘脑组织中GnIH mRNA表达水平极显著低于(P0.01)溆浦鹅,而两种鹅之间的GnIH激素在各个时期均无显著差异。这提示随着繁殖阶段的改变,在调控两种鹅产蛋性能的作用过程中GnRH可能较GnIH发挥着更为重要的作用,为进一步研究鹅繁殖分子机制奠定基础。     

鹅产蛋相关基因筛选获得成功

日前许多学者针对鹅产蛋性能普遍较低的现状,利用抑制性消减杂交技术分别构建了产蛋前期和产蛋期鹅垂体、卵巢、肝脏等组织消减cDNA文库,采用反向斑点杂交技术对文库进行了差异表达基因的筛选和鉴定,经测序获得了多个可在产蛋期鹅垂体、卵巢、肝脏组织中高效表达的基因或     

籽鹅卵巢组织差异表达基因的研究

本研究旨在筛选产蛋前期和产蛋期籽鹅卵巢组织差异表达的基因,并进行功能分析,从而探讨鹅产蛋的分子调控机理。利用抑制性消减杂交技术筛选籽鹅卵巢组织中的差异表达基因,用GOTM软件将所得ESTs从分子功能水平进行GO分类。结果表明,所构建的籽鹅卵巢组织消减cDNA文库的削减效率在20倍以上,片段大小主要在300～750bp;挑选86个阳性克隆测序,获得15个ESTs,其中有11个为已知的ESTs,4个未知的ESTs;11个已知的ESTs被分成结合功能、催化活性、酶调节活性、转运蛋白活性和其它功能。本研究成功构建了籽鹅卵巢组织消减cDNA文库,并筛选得到了15个可能在产蛋前期和产蛋期籽鹅卵巢组织中差异表达的ESTs,它们可能在鹅产蛋过程中起着重要的调控作用。     

产蛋前期和产蛋期鹅HPG轴褪黑素受体mRNA表达规律的研究

为阐明不同繁殖阶段鹅HPG轴褪黑素受体mRNA表达的变化规律,应用实时荧光定量PCR技术检测产蛋前期和产蛋期四川白鹅下丘脑、垂体、卵巢组织中3种褪黑素受体亚型(Mel-1a、Mel-1b和Mel-1c)mRNA相对表达量。结果表明:产蛋前期鹅下丘脑Mel-1a、Mel-1b和Mel-1c的相对表达量分别是产蛋期的5.25倍(P<0.01)、0.43倍(P<0.01)和1.20倍(P<0.01);产蛋前期鹅垂体Mel-1a、Mel-1b和Mel-1c的相对表达量分别是产蛋期的3.01倍(P<0.01)、7.51倍(P<0.01)、7.25倍(P<0.01);产蛋前期鹅卵巢组织中Mel-1a、Mel-1b和Mel-1c的相对表达量分别是产蛋期的1.55倍(P<0.01)、0.55倍(P<0.01)、2.86倍(P<0.01)。以上结果提示,褪黑素可通过其特异性受体的介导在HPG轴的各级水平发挥其调节作用,并且下丘脑褪黑素主要通过Mel-1a介导、垂体褪黑素主要通过Mel-1b和Mel-1c,卵巢褪黑素主要通过Mel-1a和Mel-1c介导来发挥其调控作用。     

鹅TTGGFFBBRRAAPP1基因克隆与表达分析

为探讨TGFBRAP1在不同等级卵泡和不同就巢时期卵泡中的表达规律,通过RT-PCR、RACE技术克隆TGFBRAP1基因mRNA序列,并用RT-qPCR检测浙东白鹅不同等级卵泡以及不同就巢时期卵泡的TGFBRAP1表达。结果显示:TGFBRAP1 mRNA序列长为3 870 bp,其中包括45 bp的5′UTR和1 236 bp的3′UTR和2 589 bp的开放阅读框(Open reading frame,ORF),共编码862个氨基酸;RT-qPCR检测结果显示,等级卵泡中TGFBRAP1表达量均显著高于等级前卵泡(P0.05),但均低于卵巢组织表达量,同时还发现产蛋期TGFBRAP1表达量显著高于就巢期(P0.05)。研究表明,TGFBRAP1基因与鹅生殖调控密切相关,不仅参与了鹅卵泡发育,而且参与鹅产蛋/就巢发生。     

皖西白鹅 拨出一片新天地——系列连载(五)

(上接第22期) 管理篇——休闲种鹅和公鹅的饲养管理 (一)休闲种鹅的管理 鹅的产蛋期(包括就巢期)在1年之中不足2/3，有7～8个月，还有4～5个月是休产期。当母鹅产蛋逐渐减少，每天产蛋时间推迟，小蛋、畸形蛋增多，大部分母鹅的羽毛干枯，公鹅配种能力差，种蛋受精率低，种鹅便进入持续时间较长的休产期．     

SLCO1C1和SLCO4C1基因与绿壳蛋鸡不同产蛋期壳色相关性研究

为探究SLCO家族基因与绿壳蛋壳色的关系,试验以产绿壳蛋母鸡为试验样本,分别收集产蛋前期(150日龄)和产蛋后期(500日龄)的鸡蛋、心脏、肝脏、脾脏、肌肉、卵巢、壳腺组织和血样样品,采用色差仪测定壳色数值;ELISA方法检测血清中血红素加氧酶1(HO-1)的含量;利用荧光定量PCR测定HO-1和SLCO家族基因的表达。结果显示：产蛋前期蛋壳L^*值极显著低于产蛋后期(P<0.01),而b^*值极显著高于产蛋后期(P<0.01);产蛋前期血清中HO-1含量和壳腺中HO-1表达量极显著高于产蛋后期(P<0.01);SLCO1C1和SLCO4C1在壳腺中特异性表达,SLCO1A2在肝脏中特异性表达;产蛋前期SLCO1C1和SLCO4C1 mRNA表达量显著高于产蛋后期(P<0.05)。结果表明HO-1基因表达与酶活性下降造成绿壳蛋鸡壳色变浅,SLCO1C1和SLCO4C1可能是胆绿素合成和转运的重要基因。     

种鹅产蛋高峰期与休产期繁殖生理指标的比较

为比较分析产蛋高峰期与休产期种鹅繁殖生理的差异,本试验在明确种鹅产蛋规律的基础上,分别于2月和8月随机取种母鹅15只和种公鹅10只,进行翅静脉采血,测定血液生化指标和生殖激素指标,再从中随机取种母鹅6只和种公鹅3只屠宰,取下丘脑、垂体,运用实时荧光定量PCR测定相关基因的相对表达量.结果表明,产蛋高峰期种鹅血液总蛋白、白蛋白、促黄体素和催乳素浓度以及促性腺激素释放激素基因和卵泡刺激素基因相对表达量均极显著高于休产期种鹅(P＜0.01),而尿素氮含量极显著低于休产期种鹅(P＜0.01);种母鹅碱性磷酸酶含量极显著高于休产期种母鹅(P＜0.01),血液卵泡刺激素和雌二醇含量均显著高于休产期种母鹅(P＜0.05).     

白术、微米白术和白术多糖对断奶仔猪生长性能和肠道形态及微生态区系的影响

试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

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.     

Y‐chromosomal variation of local goat breeds of Turkey close to the domestication centre

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.     

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.     

Feline intracranial neoplasia: retrospective review of 160 cases (1985-2001)

The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.