猪体细胞核移植胚体外发育初步研究

采用胞质内注射法进行猪体细胞核移植，对去核、激活和培养等关键技术过程进行研究。结果表明：（1）点压法、挤压法对卵母细胞的去核率明显高于盲吸法（三者分别为62．5％、64．6％、50．7％，P〈0．05）。对早成熟的卵母细胞（36-44h）进行去核可明显提高去核效率（P〈0．05），在36-38h、39-41h、42-44h去核率分别为60．9％、67．8％、64．3％，而45-48h为48．4％。（2）体细胞预激活有助于提高核移胚卵裂率（28．0％、20．1％，P〈0．05）。钙离子载体A23187单独或与6-二甲基氨基嘌呤（6-DMAP）联合作用能使猪体细胞核移胚激活继续发育。（3）核移胚以胚胎培养液NCSU23及卵丘单层共培养体系进行分别培养，核移胚卵裂率无明显差异（30．06％、31．5％，P〉0．05）。但NCSU23培养4细胞后发育能力更高（13．5％、3．9％，P〈0.05）。     

水牛卵母细胞去核方法的比较

水牛卵母细胞体外成熟22h后，分别用盲吸法、点击法和纺锤体成像系统（Spindle—View System）去核。结果，三者的去核率分别为65．1％、81．8％和95．0％，差异极显著（P〈0．01）。以水牛胎儿成纤维细胞为供体，通过核移植构建的重构胚的融合率、分裂率和囊胚发育率，在上述3种方法之间均没有显著差异（P〉0．05）。认为点击法是一种较为简单、实用和有效的去核方法。     

影响绒山羊母羔卵母细胞体外成熟因素的研究

研究了母羔超排方法、成熟培养时间及成熟液添加EGF对绒山羊母羔卵母细胞体外成熟的影响。结果表明,直径〉130μm组PbI率（73．27％）显著高于直径120～130μm组（P〈0．05）,直径〈120μm组PbI率（32．22％）极显著低于其他组（P〈0．01）;卵母细胞成熟培养24～27h组PbI率（68．06％）显著高于21～24h组（P〈0．05）,27～30h组PhI率（73．47％）高于24～27h组,但差异不显著（P〉0．05）;10ng／mL组和20ng／mLEGF组PhI率极显著高于对照组（P〈0．01）,20ng／mLEGF组pbI率（71．43％）显著高于10ng／mL（P〈0．05）。     

饲料中添加微生态制剂对奥尼罗非鱼生长、饲料利用和抗病力的影响

本试验采用初始体重（5．13±0．15g）的雄奥尼罗非鱼作为饲养对象．在饲料中分别添加不同水平微生态制剂（0、0．05％、0．1％、0．2％和0．3％）,共设5个处理,采用开放式流水养殖．饲养8周．研究饲料中添加不同水平微生态制剂对奥尼罗非鱼生长、饲料利用、体组成成分的影响。8周后注射嗜水气单胞菌进行感染试验．观察7d,计算存活率,研究添加微生态制剂对奥尼罗非鱼抗病力的影响。结果表明：饲料中添加微生态制剂对奥尼罗非鱼的增重率、饲料系数、特定生长率、肥满度及蛋白效率均有显著影响（P〈0．05）,但对成活率影响不显著（P〉0．05）。增重率和特定生长率．对照组均显著低于添加量在0．1％～0.3％组（P〈0．05）．与添加量为0．05％组差异不显著（P〉0．05）。饲料系数,对照组与0．05％组差异不显著（P〉O．05）,均显著高于0．1％-0．3％组（P〈0．05）。蛋白效率和肥满度,添加0．2％和0.3％组显著高于对照组和0．05％组（P〈0．05）。罗非鱼全鱼水分和蛋白含量．各组间有显著差异（P〈0．05）,而全鱼体脂肪和灰分含量,各组间差异不显著（P〉0．05）。罗非鱼攻毒24h、48h、96h和168h后,对照组存活率显著低于其他各组（P〈0．05）．但微生态制剂添加量为0．2％和0．3％时．存活率显著高于对照组和0．05％添加量组（P〈0．05）。综合增重率、特定生长率、饲料系数、蛋白质效率等生长性能指标以及攻毒后的成活率,建议在罗非鱼配合饲料中微生态制剂的适宜添加量为0．1％～0．2％。     

牛体细胞核移植技术

对牛体细胞核移植技术中核供体细胞周期同期化处理，电融合条件及去核方法等进行了研究。结果表明，血清－饥饿组（细胞周期同期化处理组）的移核重构胚细胞融合率（83．3％），卵裂率（70％）和囊胚发育率（33．8％）与血清－选择组对应指标间无显著性差异（82．3％，69．2％，32．4％，P＞0．05）；但上述2组分别极显著高于血清－随机组（64．8％，33．8％，4．35％，P＜0．01），试验确立了场强1.025kV/cm，脉冲宽度50μs，连续2次脉冲为最佳电融合条件。摸索出的点击去核法，对卵母细胞进行去核，去核成功率达90％，极显著高地吸引法的68．3％（P＜0．01）；其囊胚发育率为34．1％，显著高于吸引法的18．0％（P＜0．05），与挤压法（20．9％）相比差异不显著（P＞0．05），移植重构胚胎移植后，产2头成活的体细胞克隆牛。     

牛、羊卵母细胞孤雌激活的研究

本试验主要比较了离子霉素、电脉冲两种方法激活牛、羊体外成熟卵母细胞的效率。两种激活方法中。牛胚胎卵裂率无显著差异（90．61％对94．40％，P〉0．05），而离子霉素激活胚胎的囊胚发育率极显著高于电激活方法（12．3％对2．4％，P〈0．01）。两种方法对羊胚胎的研究中，羊胚胎卵裂率无显著差异（72．4％对77．4％，P〉0．05）。但是离子霉素激活胚胎的囊胚发育率显著高于电激活方法（3．67％对10．40％，P〈0．05）。本试验中还比较了用化学激活法（离子霉素）激活牛体外成熟卵母细胞后，用SOFaa体系培养，换液与不换液对孤雌激活胚胎体外发育的影响。结果表明：在第4天不换液的胚胎卵裂率和囊胚率极显著高于换液的胚胎（11．64％对3．49％。P〈0．01）。     

三种同期发情方案对奶牛胚胎移植效果的影响

同期发情是胚胎移植的关键技术环节，本试验通过三种不同的同期发情方案比较其对奶牛胚胎移植效果的影响。结果显示：CIDR＋PG法同期发情率高于直检后一次PG法和2次PG法，但差异不显著（P〉0．05）。可是其48h内的同期化程度显著高于另外两组方案（P〈0．05）。三种方案的黄体合格率、妊娠率均无显著差异（P〉0．05），2次PG法高于另外两组。受体牛的初选合格率育成牛（87．13％）显著高于成母牛（68．5％）（P〈0．05）。育成牛同期发情率为89．04％，高于成母牛的77．01％，黄体合格率、妊娠率育成牛均略高于成母牛，但统计学上差异均不显著（P〉0．05）。     

猪卵巢卵母细胞采集方法的比较研究

本试验利用抽吸法和剖切法两种采集方法采集猪卵巢卵母细胞,对获得可用卵丘卵母细胞数和卵丘完整程度的影响进行比较研究,并分析在卵巢收集时不同的保存温度、运输时间对卵母细胞体外成熟的影响。结果表明：剖切法采集细胞数目显著高于抽吸法（P〈0．01）;A、B级卵母细胞的比例在剖切法中显著高于抽吸法（P〈0．05）;抽吸法所采集的c级卵母细胞显著高于剖切法（P〈0．05）;剖切法中卵母细胞卵丘完整率高于抽吸法（P〈O．05）;离体卵巢保存的最佳温度为36℃-38℃;其保存时间为1～1．5h时成熟率57．6％与2—2．5h时成熟率52．8％相比无显著差异（P〉O．05）;保存时间超过6h时卵母细胞的成熟率显著下降。     

多山梨醇酯20对棉花秸秆体外发酵的影响

采用体外培养技术，研究不同添加水平的多山梨醇酯20对棉花秸秆体外发酵的影响。结果表明：多山梨醇酯20能显著提高棉花秸秆体外发酵24h的累积产气量和发酵速度。与对照组相比，0．1％、0．2％、0．4％和0．8％的多山梨醇酯20使24h累积产气量分别提高27．95％（P〈0．01）、32．30％（P〈0．01）、43．48％（P〈0．01）和19．88％（P〈0．01）。多山梨醇酯20使棉花秸秆中的干物质、有机物、纤维素和半纤维素24h降解率提高，其中0．2％组的干物质、有机物和半纤维素的24h降解率为各组最高，分别比对照组提高12．71％（P〈0．05）、11．81％（P〉0．05）和115．96％（P〈0．01）；纤维素的24h降解率以0．4％最高，较0％提高33．67％（P〈0．01）。添加多山梨醇酯20后，使pH和还原糖显著下降。除0．2％组外，总挥发性脂肪酸显著高于对照组。羧甲基纤维素酶（CMCase）活性有增加的趋势，0．8％组的CMCase活性较0％组高114．05％（P〈0．01）。低浓度多山梨醇酯20有提高微生物（MCP）的趋势，但随着多山梨醇酯20添加浓度的增加，MCP逐渐下降，培养上清中的氨态氮浓度呈现线性增加。在体外厌氧培养条件下，多山梨醇酯20明显促进棉花秸秆在混合绵羊瘤胃液中的降解。     

不同促性腺激素添加方式对猪卵母细胞的影响

在卵母细胞体外成熟培养过程中，培养基中添加激素与否及其激素添加的先后顺序是影响猪卵母细胞核成熟和质成熟的一个重要因素。本试验将猪卵母细胞分别在FSH→不含激素、FSH→LH、FSH＋LH→不含激素中培养48h（培养第20～22h后换液），并于成熟培养的第24h（未换液）、48h将卵母细胞进行荧光染色，观察其生发泡内染色质构型及卵母细胞核成熟情况。实验表明：1）在IVM的前24h，添加FSH＋LH组的GV Ⅳ期卵母细胞比例低于只添加FSH组，但差异不显著（8．99％vs17．19％，P〉0．05）；2）在FSH存在的情况下，IVM的前期和后期添加LH能促进卵母细胞发生GVBD；3）FSH＋LH培养24h后转入不含激素培养基组，卵母细胞的核成熟比率显著高于添加FSH组和先添加FSH培养24h后转入添加LH组（64．36％vs20．45％、36．62％、41．98％，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。     

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.     

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.     

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.     

The Prevalence and Intensity of Helminth and Coccidial Infections in Dairy Cattle in Central Kenya

A survey of gastrointestinal parasite infections of young (<6 months old), immature (6–12 months old) and adult (>12 months old) dairy cattle on 16 farms in Kiambu District, Kenya was conducted during a dry season (September 1991 to January 1992) and during a wet season (March to July 1992). The survey was based on monthly coproparasitological examination of cohorts and worm counts in tracer calves. The effects of age, sex, farm and season on the prevalence and intensity of helminth and coccidial infections were determined. Faecal egg and oocyst counts revealed that the overall prevalences were: strongyles (including trichostrongyles) (85.5%), liver flukes (Fasciola gigantica) (34.0%), coccidia (30.9%) and tapeworms (9.6%). Eight species of the protozoan Eimeria were identified, the most prevalent species being E. bovis and E. zuernii. The most prevalent nematode genera were Haemonchus, Cooperia, Oesophagostomum and Trichostrongylus. Season, farm and age of the animals had a significant (p<0.05) influence on the intensity of infection with strongyles, liver flukes and coccidia, whereas the sex of the animals had no significant (p>0.05) effect on the prevalence or intensity of infections. A higher intensity of infection with strongyles and coccidia was found in the wet season than in the dry season (p<0.05). The age-specific intensity was in the following order: for strongyles, immature animals of 6–12 months of age had the highest egg counts, followed by young calves and adults. Calves had significantly (p<0.05) higher oocyst counts than immatures or adults. Liver fluke egg counts did not differ significant (p>0.05) between immatures and adult cattle.     

A brief history of the prevention of infectious diseases by immunisations

Infectious diseases have always been a terrible scourge for humans. The appearance of these plagues, as they were called without distinction, was generally connected to various conditions: asters, climatic changes or religious reasons. The concept of contagious, and then infectious, diseases came slowly. Variolation, i.e. transmission of ‘virulent’ matter to induce a natural disease and the immunity against it, was brought from Constantinople to England by Lady Montague, in 1721. This ‘variolation’ technique was also often performed in veterinary medicine against diseases like sheep-pox or pleuropneumonia. As ‘vaccination’ is the term generally accepted for ‘immunisation’, variolation can be the word designating such a technique. The second period of the history of immunisation began, in 1880, with the studies of Pasteur and his collaborators. A great number of bacterial vaccines were developed: dead, live but attenuated or only parts of pathogens. The viruses were produced in animals, then in eggs and at last, in tissue cultures. Second generation vaccines appeared with genetic engineering: recombinant vaccines, vector vaccines, nucleic acids vaccines, and markers vaccines, among others. These novel technologies can permit the development of new ones and improve the quality of the vaccines already existing.     

Micronutrient Content of Certain Tropical Conventional and Unconventional Feed Resources of Southern India

The ash, silica and certain important micronutrients were estimated in conventional and unconventional feed and fodder resources available in Southern India. Commonly used dry roughages, such as paddy straw, ragi straw, maize kadbi/stalk, jowar kadbi/stalk, bajra stalk and wheat straw, were high in ash (9.9% +/- 0.77%) and silica (6.4% +/- 0.65%) and low in most of the other micronutrients, except iron, with paddy straw containing most silica (>9%). Cultivated non-leguminous (maize, jowar) and improved green crops (hybrid napier, guinea, green panic, NB-21, CO-1) were also high in ash (10.5% +/- 0.60% and 12.5% +/- 0.51%) but were moderate sources of P, Mg and Cu and good sources of Zn (98 +/- 13.8 ppm and 55 +/- 6.7 ppm). Leguminous green fodders (stylosanthus, lucerne, cow pea, soyabean) were excellent sources of Ca (1.9% +/- 0.16%), Mg (0.40% +/- 0.05%), Cu (30 +/- 5.2 ppm), Zn (121 +/- 14.7 ppm) and Fe (1234 +/- 166 ppm) and moderate sources of P. Mixed local grasses and weeds were high in silica (6.9% +/- 1.00%) but were good sources of Cu, Zn and Fe. Cereal grains (maize, wheat, rice, ragi) were low in ash (2.9% +/- 0.33%) and were relatively poor sources of Ca (0.22% +/- 0.03%), Mg (0.19% +/- 0.03%) and Cu (13 +/- 3.1 ppm). Pulses were low to medium sources of most minerals and good sources of Fe (1230 +/- 293 ppm). Oil seed cake/extractions (groundnut cake, cotton seed cake, soyabean meal, sunflower cake, safflower cake) and cereal by-products (rice polish, rice bran, wheat bran) were excellent sources of P (1.1% +/- 0.47% and 2.3% +/- 0.19%) and good sources of Zn (65 +/- 3.9 ppm and 66 +/- 10.7 ppm) and Fe (938 +/- 130 ppm and 662 +/- 126 ppm). Among the unconventional feeds screened, orange peel, sunflower heads, meat meal, rubber seed cake, spirulina algae and sea weeds contained plentiful Ca, Zn and Fe: tree leaves/top feeds (mulberry, erythrina, glyricidia, banana, subabul, groundnut haulms) were excellent sources of Ca (1.5% +/- 0.13%), Zn (120 +/- 22.9 ppm) and Fe (1033 +/- 133 ppm) but relatively poor sources of P. Soyabean husk, cocoa seed husk, rubber seed cake and meat meal were moderate to good sources of P (1.0% and 0.33%). The high Zn and Fe values of most feeds/fodders were probably due to soil contamination. This account of the micronutrient content of feed/fodder resources should help in strategic supplementation intended to alleviate local deficiencies.     

Bovine Mastitis in Selected Areas of Southern Ethiopia

A study on bovine mastitis, designed to determine the causal agents, prevalence of infection and impact of risk factors in three cattle breeds, was conducted in selected areas of southern Ethiopia. A total of 307 lactating and non-lactating cows, of which 162 were indigenous Zebu, 85 Jersey and 60 Holstein-Friesian, were examined by clinical examination and the California mastitis (CMT) test. Of these, 40.4% were positive by CMT and bacteriology for clinical or subclinical mastitis, with prevalence rates of 37.1% and 62.9%, respectively. Out of 1133 quarters examined, 212 (18.7%) were found to be infected, 83 (39.2%) clinically and 129 (60.8%) subclinically. The prevalence of mastitis was significantly higher in Holstein-Friesian than in indigenous Zebu, in non-lactating cows than in lactating cows, in the early lactation stage than in the mid-lactation stage, in cows with lesions and/or tick infestation on skin of udder and/or teats than in cows without this factor, and in the wet season than in the dry season. Mastitis increased with parity number (R = 0.9). Of 248 CMT and clinically positive udder quarter samples analysed microbiologically, 212 were culturally positive for known mastitis pathogens and 36 were negative. Of the 199 positive samples, Staphylococcus accounted for 39.2%, Streptococcus for 23.6%, coliforms for 14.1%, Micrococcus and Bacillus species for 8.0% each and Actinomyces or Arcanobacterium (Corynebacterium) for 7.0%. It was concluded that there was a high prevalence of clinical and subclinical mastitis, mainly caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli, in this study area.     

不同日粮氮水平对山羊氮代谢和微生物蛋白质合成的影响

本试验旨在探讨不同日粮氮水平对内蒙古白绒山羊氮代谢和微生物蛋白质合成(MCP)的影响,以达到提高氮利用率,减少环境污染和饲料资源的浪费。试验选用9只体况良好、体重为(43.83±2.95)kg,装有瘤胃瘘管的内蒙古白绒山羊,按体重随机分为3组,每组3只。日粮分为低氮7.5%、中氮10.5%、高氮13.5%3个氮水平。试验使用常规试验方法分析测定粪尿血液等生化指标,通过全收尿法和尿嘌呤衍生物法估测MCP。结果表明:随着氮水平增加,氨氮(NH3-N)浓度、氮摄入量、尿氮排出、尿中尿素氮(UUN)和MCP显著增加(P0.05),干物质采食量(DMI)显著降低(P0.01);沉积氮占总摄入氮的比例在中氮组最高,MCP在高氮组最高。综上所述,适当降低日粮氮水平可减少粪尿氮排放,提高反刍动物氮利用率。