THE EXPERIMENTAL TRANSMISSION OF PINTA,MAL DEL PINTO OR CARATE TO THE RABBIT

In this preliminary note we describe the results obtained by inoculation of the skin of the scrotum of four black rabbits with serous fluid expressed from an initial pinta lesion. The inoculation was made intradermally, as superficially as possible, with serous fluid rich in treponemata and diluted with normal salt solution. A papule appeared at the point of inoculation in one of the four rabbits on the 105th day. One hundred and fifteen days after inoculation the papule presented a circular erosion on an indurated base. The serous fluid expressed from this lesion contained numerous treponemata on dark-field examination, and these were easily impregnated by the Fontana-Tribondeau method. A volunteer inoculated with serous fluid expressed from the rabbit's lesion developed a typical initial pinta lesion at the point of inoculation, thus proving that the scrotal lesion of the rabbit was produced by Treponema carateum or Treponema herrejoni, the causal agent of pinta, mal del pinto or carate. Four rabbits inoculated with serous fluid from the scrotal lesion of the rabbit have not yet developed visible lesions.     

一种快速鉴定柑桔体细胞杂种对脚腐病抗性的方法

通过用菌丝块接种柑桔叶片法进行柑桔体细胞杂种抗病性鉴定,并与接种柑桔枝条法,菌丝块接种嫁接苗法比较,证明柑桔不同部位对脚腐病的抗性存在着相关性,可采用这种相关性接种柑桔叶片进行体细胞杂种的抗病性鉴定。     

兔出血症病毒试验感染及临床病理学观察

以不同日龄及生理状态下的中国白兔为试验动物,经静脉接种、皮下接种、滴鼻和同居感染兔出血症病毒,进行临床检查和病理学观察。结果表明,静脉接种感染的兔潜伏期比其他途径感染的兔短;2月龄以下幼兔有明显的抵抗力,其他月龄的兔均易感,怀孕兔全部发生流产;临床病理学观察以各实质器官的退行性病变及弥散性血管内凝血为主要特征,表现为各组织器官严重出血、水肿,实质细胞广泛性变性、坏死,微血管内大量血栓形成、炎性细胞浸润等。     

兔出血症病毒衣壳蛋白重组腺病毒的构建、表达及其免疫原性

通过RT-PCR方法扩增上海地区兔出血症病毒(RHDV)衣壳蛋白基因(VP60).将其克隆入腺病毒表达载体系统中,构建成重组腺病毒质粒,用脂质体法转染HEK293细胞,得到重组病毒pAd-VP60,经RT-PCR、IFA、HA、SDS-PAGE和Western-blotting鉴定,结果表明重组VP60蛋白在HEK293细胞中获得表达.表达产物与铝胶佐剂混合后注射3月龄非免疫健康家兔,结果表明,免疫后21 d兔体内产生抗RHDV抗体,其HI效价可达2~6～2~7,能抵御HA≥2~(10)致死剂量RHDV强毒的攻击,保护率为100%.     

rhPA/GH双转基因兔的制备及rhPA表达检测

【目的】双基因共整合可协同促进转基因生物的目的基因表达水平提高,研究获得rhPA/GH双转基因兔,并比较分析目的基因rhPA的表达水平及兔个体生长发育情况,为制备高表达rhPA转基因动物和遗传育种提供新思路。 【方法】利用NotⅠ/SalⅠ双酶切PCL25/GH质粒和QIAGEN DNA胶纯化试剂盒回收显微注射用基因片段。以3只rhPA单转基因兔（标号K06、K10、K17）作为供体兔,通过FSH/hCG超数排卵、受精卵原核显微注射、胚胎移植、苯酚/氯仿抽提新生仔兔耳尖组织基因组、PCR整合检测等方法获得rhPA/GH双转基因兔。经ELISA和Western blotting对转基因兔乳清进行表达检测,比较分析单、双基因表达rhPA水平。测量不同月龄的转基因兔体重,通过监测双转基因兔不同生长阶段的体重来分析GH对兔生长发育的影响。【结果】成功获得了约16 700 bp 大小的显微注射用基因片段,共超排3只供体兔获得122枚卵,其中103枚受精卵,受精率为84.4%（103/122）,显微注射后挑取形态较好的81枚移植6只同步发情受体兔,5只兔怀孕,妊娠率为83.3%（5/6）,妊娠到期共出生32只仔兔。通过PCR检测鉴定有19只携带rhPA基因的转基因兔,其中有11只rhPA/GH双转基因兔（7♂,4♀）,双基因整合率为34.4%（11/32）,且4只rhPA/GH双转基因母兔来源亲代分别为K06供体兔2只（标号K06-1、K06-2）、K10供体兔1只（标号K10-1）、K17供体兔1只（标号K17-1）。K06号rhPA单转基因兔乳清中rhPA表达量为42.2μg·mL^-1,K06-1和K06-2号rhPA/GH双转基因兔乳清中rhPA表达量分别为432、444μg·mL^-1;K10号rhPA单转基因兔乳清中rhPA表达量为42.8μg·mL^-1,K10-1号rhPA/GH双转基因兔乳清中rhPA表达量为636μg·mL^-1;K17号rhPA单转基因兔乳清中rhPA表达量为15.2 μg·mL ^-1,K17-1号rhPA/GH双转基因兔乳清中rhPA表达量为248 μg·mL ^-1。4只rhPA/GH双转基因母兔（K06-1、K06-2、K10-1、K17-1）乳腺表达rhPA含量在248-636μg·mL^-1之间,远远高于rhPA单转基因母兔（K06、K10、K17）乳腺的表达含量（15.2-42.8μg·mL^-1）,表达水平显著提高了约10.2-16.3倍,说明GH能够协同促进目的基因rhPA在转基因兔乳腺中的表达水平。Western blotting结果显示出现一约39.0 kD大小的条带,与目的蛋白rhPA大小相同,进一步证明转基因兔乳腺中表达的这种蛋白为目标产物rhPA。对rhPA/GH双转基因兔从出生到6个月的体重进行连续测量,发现与正常非转基因兔的体重没有明显的差异,绘制生长曲线进一步表明4只整合GH的转基因兔与2只未整合GH的正常兔在生长发育的不同阶段体重上没有显著性差异,成长至6个月的体重均在4.0-5.0 kg之间,这证明GH的导入并不影响转基因兔的存活和正常生长发育至成年。【结论】成功地制备了rhPA/GH双转基因兔,并证明了GH基因的导入能够显著地提高目的基因rhPA的表达量,且不会对转基因兔的生长发育产生影响,这为将来制备高表达转基因兔及其它动物奠定了基础,也为转基因动物乳腺生物反应器和转基因育种建立提供了新思路、新方法。     

家兔A型魏氏梭菌病病因及防治报告

本文通过对河北省A、B两兔场暴发家兔A型魏氏梭菌病病因的调查分析和治疗试验,得出如下结论:(1)该病诱因是饲料营养失调,高能量、高蛋白、低纤维,速成后肠碳水化合物过度负荷所致。长途运输、饲料更换、冷潮刺激等也是该病的诱因。(2)该病死因主要是由于魏氏梭菌外毒素导致神经系统中毒、肠道粘膜严重损伤和极度脱水所致。(3)该病主要症状是急性水样腹泻,体温正常或偏低。病变主要是肓肠出血和胃粘膜溃疡。(4)采取大剂量口服青、链霉素、补液、解毒和收敛等综合措施,可控制病情发展,并使轻症群治愈率76.67%,重症群52.63%,病危群9.38%。     

家兔病原性大肠艾希氏杆菌的致病力检验

应用分离于自然家兔大肠杆菌病例的O_(20)、O_7、O_(70)、O_(21)和_(132)等5个不同血清型代表株进行了对不同日龄兔的发病试验,表明均具有一定的致病力,不同日龄兔均可被感染发病或死亡,不同接种途径感染试验表明以静脉接种、肌肉注射、胃内接种和皮下注射均可引起感染,尤以静脉接种最易导致发病及死亡,不同型别株间对比以O_(20)毒力最强,被感染发病死亡兔剖检病变与自然病例基本一致,且均能回收到原攻毒菌,耐过兔多数出现不同滴度的相应血清抗体。     

辣椒炭疽病抗性鉴定方法研究

比较了离体果实接种和苗期喷雾接种两种方法在辣椒炭疽病抗性鉴定中的效果。苗期喷雾接种,供试8个品种发病率在3.0%~28.4%、病情指数在0.8~12.5之间,品种之间差异不显著,无法区分抗、感病品种间的抗病性差异。而离体果实接种发病率在0%~92.5%,抗病品种发病轻,果实上无病斑或只形成一些小病斑,没有孢子产生;感病品种发病重,果实上形成典型的凹陷病斑,并产生大量的橙色孢子;抗、感病品种之间的发病程度差异显著。结果表明,离体果实接种法更适合用于辣椒炭疽病抗病性鉴定。     

貉源李氏杆菌的分离与鉴定

采用常规细菌分离方法从病死貉的脑、肝脏、肾脏分离培养得到6株菌株,对分离菌株进行形态特征、培养特性观察及生化特性鉴定,结果确诊分离菌株为李氏杆菌。以该菌株接种家兔,接种后第7d开始出现死亡,并从家兔的肝脏、脾脏检测出与接种细菌一致的菌株,证实了李氏杆菌对家兔亦有一定的致病性。药敏试验结果表明,该分离菌对卡那霉素、头孢噻肟钠、万古霉素、庆大霉素、丁胺卡那霉素、氨苄青霉素、链霉素敏感;必要时应筛选出具有协同作用和累加作用的抗生素,以便临床上联合应用抗生素,提高治疗李氏杆菌病的疗效。     

应用协同凝集试验诊断家兔和牛霉形体感染

以兔抗牛肺疫 C88004株血清制备5%协同凝集试验诊断液并用之于检测兔和牛霉形体感染,结果表明:4只人工感染 C88004株兔肺与气管浸出液阳性检出率分别为75%(3/4)和100%(4/4),20只教学用兔分别为20%(4/20)和0%,6只兔病毒性出血症病毒感染兔皆为阴性.采自牛肺疫疫区14例牛肺浸出液和胸水的阳性检出率分别为78.6%(11/14)和21.4%(3/14)。协同凝集试验用于检测牛肺疫病原,特异性强,重复性高,简便易行,值得推广。     

兔出血症病毒主要结构多肽的纯化及免疫保护性试验

人工感染ＲＨＤＶ－ＮＪ株病死兔肝经－５０℃反复冻融,氯仿处理,ＰＥＧ沉淀、差速离心,Ｓｅｐｈａｒｏｓｅ－４Ｂ柱层析得纯化病毒,纯化ＲＨＤＶ经ＳＤＳ－ＰＡＧＥ、ＣｕＣｌ２负染色后,切取主要结构多肽ＶＰ１（６４．５ＫＤ）凝胶带,除去ＣｕＣｌ２和ＳＤＳ获得ＶＰ１,高效液相色谱鉴定只有一个蛋白峰,Ｗｅｓｔｅｒｎ－Ｂｌｏｔｔｉｎｇ,ＨＡ和ＨＩ结果显示纯化ＶＰ１具有良好的抗原表位稳定性。无母源抗体的３月龄兔１     

Nature of "sex-ratio" agent in Drosophila

Several lines of evidence implicate small spirochetes, presumably treponemata, as etiologic agents in the production of the maternally transmitted "sex ratio" condition (SR) in Drosophila nebulosa, in D. willistoni, and in strains of D. melanogaster into which the SR condition has been artificially transferred. The presence of treponemata in the hemolymph of adult females of these species is completely correlated with the production of unisexual progenies and like this condition is dependent on the genotype of the host and of the infectious agent.     

Detection of a gonadotropin in rabbit blastocyst before implantation

A gonadotropin similar to human chorionic gonadotropin or luteinizing hormone has been demonstrated in rabbit blastocyst prior to implantation. The gonadotropin has been detected by a radioreceptor assay for human chorionic gonadotropin with the use of the plasma membranes of bovine corpora lutea obtained during the first trimester of pregnancy. The concentrations of the human chorionic gonadotropin or luteinizing hormone per milliliter of blastocyst fluid were tenfold higher than those in the blood of pregnant rabbits on days 5 and 6 after mating.     

幼兔饲养中影响成活和增重主要因素的探讨

 养兔具有投资少、效益高、见效快等优点。兔养殖中存在的主要问题是幼兔死亡率高。山西省幼兔平均成活率为57%，全国兔的死亡率为30%-50%，严重影响养兔业的发展。通过日喂量测定和兔出血症感染试验，探讨出精料、青草日喂量和兔出血症感染是幼兔饲养中影响成活和增重的主要因素。一日内的精料喂量以占体重的2.5%,消化能0.322兆焦/公斤体重、食入粗蛋白质3.90克/公斤体重,青草(含干物质30%)25%,消化能0.912兆焦/公斤体重、食入粗蛋白质3.10克/公斤体重为适宜。这样，幼兔成活率可达到90%以上，增重相对提高23.3%,差异显著(P<0.05),料重比最低。日喂量如果过大，会引起消化不良，造成营养物在盲肠等大肠内的积聚，使腹泻和腹胀等消化道病增多，成活率降到60%-75%。60日龄的幼兔即可感染兔出血症（RHD），感染死亡率（18.5%-66.7%）与日龄呈正相关,r=0.9698,P<0.01,相关关系极显著。在幼兔饲养中做到料、草日喂量适宜，断奶与防疫同步，对提高成活率和增重 具有重要意义。     

醉马草内生真菌对家兔的影响

通过饲喂试验,研究了带内生真菌(E+)和不带内生真菌(E-)的醉马草对家兔健康的影响。内生真菌的侵 染,使醉马草粗脂肪含量显著提高,粗蛋白和粗纤维含量无显著改变。按10 mL/kg体重的剂量灌服E+醉 马草水提液,家兔表现出明显的中毒症状;但累计灌服20 mL/kg体重的剂量,亦未引致死亡。平均按每 kg体重累计饲喂E+醉马草131.6 g,可引起家兔中毒;累计饲喂350.9 g可使家兔致死,死亡率达66.7%。 而灌服和饲喂E-醉马草的家兔则无异常。初步认为,醉马草对家兔的致毒机理是由内生真菌与宿主互作 产生的生物碱所致。     

兔轮状病毒性腹泻的流行病学和病原学研究

 1990～1991年,作者对云南一些兔场的幼兔腹泻进行了流行病学调查、病原学和血清学检验,成功地从腹泻死亡幼兔肠内容物中分离出2株兔轮状病毒,经形态学、生物学、血清学、核酸分析以及本动物回归等方面的鉴定,确定为A群血清3型轮状病毒.据此确立了幼兔轮状病毒性腹泻在我国的存在.     

对兔疥螨病的临床防治对策

1984～1988年武威市13群1488只家兔中发生疥螨病的446只,发病率29.97％。药物彻底治疗256只,治愈率100％;未彻底治疗190只全部死亡。采取药物治疗和环境灭虫的综合措施,才能获得良好的防治效果。     

Enhancement of the Immune Response to Rabbit Hemorrhagic Disease Vaccine in Young Rabbits by Advanced Vaccination and Chinese Herbal Adjuvants

Experiments were conducted to determine the effects of advanced vaccination and Chinese herbal adjuvants （CHA）, containing astragalus polysaccharides （APS） and ginsenosides （GS） on the immune response to rabbit hemorrhagic disease （RHD） vaccine in young rabbits. In experiment 1, 5 New Zealand rabbits of each group at 30, 35, 40, or 45 days of age were injected with 2 mL of inactivated RHD vaccine, respectively. The dynamic changes of antibody titers were tested by the hemagglutination inhibition （HI） method. In experiment 2, 30 New Zealand rabbits at 35 days of age were randomly assigned to 5 treatment groups, representing inoculation with 3 mL of non-adjuvant RHD vaccine, CHA-RHD vaccine, CHA-HA vaccine （half dose antigen）, aluminium adjuvant-RHD vaccine, and PBS, respectively. The dynamic changes of peripheral lymphocyte proliferation and serum antibody titers were tested by the MTT method and the HI method. The results showed that the titer of maternal HI antibody in the 35-day-old rabbits was lower than the protective level of 3 log2, while on days 7 to 49 after the vaccination, the antibody titers were higher than 3 log2. The CHA promoted the lymphocyte proliferation and enhanced the serum antibody titer （P〈 0.05）. These findings from the two experiments suggested that advanced vaccination and Chinese herbal adjuvants significantly enhanced the immune response to vaccine against RHD, and effectively protected the young rabbits against RHD challenge.     

PATHOGENIC PLEUROPNEUMONIA-LIKE MICROORGANISMS FROM ACUTE RHEUMATIC EXUDATES AND TISSUES

In suitable cell-free media it has been possible to cultivate pleuropneumonia-like microorganisms from the following materials, first, from chorioallantoic membranes in which lesions were apparently induced by exudates from patients with rheumatic fever; second, from pneumonic lungs of mice inoculated with similar exudates or with suspensions of the abovementioned abnormal membranes; and third, directly from the arthritic exudate of a patient with rheumatic fever, and also from an erythema nodosum nodule excised from a patient with this same disease. With three different subcultures from joint fluid, iritis has been induced in rabbits; and following intranasal inoculation with the same cultures there has developed in mice a pneumonia similar to that found in mice inoculated with rheumatic exudates and with suspensions of chorioallantoic membranes infected with rheumatic exudates. Therefore it seems probable that in all instances the pathogenic agent was derived from similar sources, viz., patients with rheumatic fever. Further work will be required to demonstrate the etiologic significance of these pathogenic agents in rheumatic fever.     

饲粮添加油橄榄叶对育肥期獭兔生长、 血液生化指标及抗氧化能力的影响

【目的】研究日粮中添加不同水平油橄榄叶对育肥期獭兔生长、血液生化指标及抗氧化能力的影响,为獭兔生产中合理添加油橄榄叶提供理论依据。【方法】将2月龄体重相似的128只生长獭兔随机分成4组,每组8个重复,在基础日粮中添加不同水平的油橄榄鲜叶（A0=0%、A1=5%、A2=10%和A3=15%）,预试期7 d,正试期38 d。【结果】①饲喂不同比例的油橄榄叶对獭兔的采食无显著影响;②添加10%和15%油橄榄叶獭兔平均日增重显著高于对照组和5%水平组（P＜0.05）,平均日增重提高7.60%;③添加油橄榄叶对葡萄糖、总蛋白、白蛋白、球蛋白和血红蛋白含量及红细胞数、中性粒细胞数等无显著影响,而对尿素氮、肌酐、白细胞数和淋巴细胞数影响显著（P＜0.05）,但在正常生理值范围内;④添加10%—15%油橄榄叶对血液的抗氧化酶活性的影响显著（P<0.05）。【结论】在饲粮中适宜添加油橄榄叶不影响育肥獭兔的生产性能,不会对血液生化指标产生负面影响,添加10%—15%的油橄榄叶能增强獭兔血液的抗氧化力。