从野外昆虫体内分离的5株微孢子虫的生物学特性与分子系统发育研究

从广西蚕区的野外昆虫菜粉蝶(Pieris rapae)、桑尺蠖(Hemerophila atrilineata)、斜纹夜蛾(Spodoptera litura)的体内收集到5株对家蚕有一定食下感染力的微孢子虫,通过生物学性状、分子系统发育研究,为其分类鉴定提供依据。来自野外昆虫的5株微孢子虫均具有Nosema属的生物学特性:显微镜下观察形状均呈长卵圆形,与家蚕微孢子虫(Nosema bombycis,Nb)有较明显差异;在生活史的各发育阶段均为双核,以二分裂方式增殖,最后形成双核成熟孢子;与Nb抗血清均产生阳性凝聚反应;超微结构观察均具双核,极丝圈数在12～14圈之间,极丝倾斜角与Nb有差异。根据5株野外昆虫微孢子虫与其它昆虫微孢子虫基于SSU rRNA基因序列构建的系统发育树,以及基于SSU rRNA和rRNA ITS基因序列进行的相似性和遗传距离分析,初步认为5株野外昆虫微孢子虫与家蚕微孢子虫同属异种。     

一株家蚕病原性微孢子虫的生物学特性与分子系统发育分析

从家蚕体内分离得到一株新的病原性微孢子虫,编号为GXM1。光学显微镜下观察GXM1微孢子虫为长卵圆形,大小(1.85±0.15)μm×(4.19±0.18)μm,在生活史的各发育阶段均为双核,以二分裂方式增殖,发育速度缓慢,发育周期约8～10 d。GXM1微孢子虫与家蚕微孢子虫(Nb)的抗血清产生阳性凝聚反应。利用透射电子显微镜观察到GXM1微孢子虫的超微结构具双核,极丝11～12圈,极丝倾斜角约45°。以上生物学性状显示GXM1微孢子虫具有微孢子虫属(Nosema)的基本分类特征。依据GXM1微孢子虫与其它昆虫微孢子虫的SSU rRNA基因序列构建的系统发育树,以及序列相似性和遗传距离分析,进一步证实GXM1微孢子虫属于Nosema属。GXM1微孢子虫对蚁蚕的半数感染浓度(IC50)为6.06×105mL-1,对家蚕的胚种传染率可达23.28%,是一株具有较强致病性和危害性的家蚕病原性微孢子虫。     

家蚕分离的微孢子虫Vairimorphasp.MG_4的研究

从家蚕分离到一种小型微孢子虫 (MG4 )。MG4 孢子大小为 2 72± 0 1 2 μm× 1 98±0 2 2 μm ;孢子具单核、双核 2种类型 ,极丝在 6～ 9圈之间 ;在家蚕体内以 2种不同的生活史发育 ,发育过程符合变态孢虫属Vairimorpha微孢子虫发育特征 ;对家蚕有弱的食下传染能力 ,主要寄生在后部中肠的气管丛 ;感染家蚕后未见胚种传染     

从东方粉蝶分离的一种变态微孢子虫的研究

从东方粉蝶成虫体中分离到一种微孢子虫 (M Pc1) ,其孢子大小为 3 2 7± 0 36 μm× 1 6 8± 0 16 μm ,孢子具单、双核两种类型 ,在家蚕体内可见到Nosema和Thelohania两种不同类型的发育过程 ,认为M Pc1应归入Vairi morpha属。这种微孢子虫对家蚕具有很强的食下感染能力 ,并有弱的胚种感染率     

2种野外昆虫来源微孢子虫的超微结构及生活史观察

将从湖南蚕区野外昆虫菜粉蝶(Pieris rapae)、桑尺蠖(Phthonandria atrilineata)分离到的2种微孢子虫感染家蚕,观察微孢子虫的寄生组织、生活史以及原代微孢子虫与在蚕体侵染繁殖的第1代微孢子虫的超微结构,作为探讨家蚕微粒子病发生与野外昆虫微孢子虫的关系的病原生物学依据之一。用Giemsa染色镜检观察的结果显示,2种来源的微孢子虫可在家蚕幼虫体内全面寄生,在中肠、丝腺、马氏管、脂肪体、生殖腺、肌肉组织、气管丛等组织中均能检出。电子显微镜观察2种来源的微孢子虫的超微结构,并与家蚕微孢子虫(Nosema bombycis,Nb)比较,结果显示在家蚕体内继代的源自菜粉蝶和桑尺蠖的微孢子虫的超微结构特征与Nb基本一致,孢子壁由3层组成,极体分为前后2部分,极丝同型,核成对出现,核形不规则,核膜双层,粗面内质网附含大量的核糖体,孢子后部有后极泡。2种来源的微孢子虫在家蚕体内的发育过程较为相似,所有时期的核都成对出现,以二分裂方式增殖,发育周期分别为72、96 h。     

山东省病原性家蚕微孢子虫的初步研究

从山东省境内家蚕成虫体内分离到 4种病原性家蚕微孢子虫 ,与N .b比较研究结果表明 ,4种微孢子虫的大小、形态、极丝圈数及倾斜角与N .b相比均存在差异 ,但血清学反应及在蚕体内的发育过程与N .b相同。初步认为该 4种病原性微孢子虫为N .b的同属微孢子虫。这 4种微孢子虫对家蚕具有极强的食下感染率和胚种传染力。     

家蚕幼虫体内微孢子虫(Nosema bombycis)两种类型孢子的形成

著者等报告了从一种夜蛾(Spodoptera depravate Butler)的微孢子虫(Nosema sp.)在昆虫组织培养细胞内发育的过种中,当所谓成熟孢子形成之前形成极丝卷曲数少的异型孢子,此类孢子能自行发芽并在细胞内建立感染且能持续进行(Iwano & Ishihara,1989)。其后又报告了家蚕微粒子病病原(Nosema bombycis)在同一培养细胞内与Nosema sp.相同的也在形态(极丝卷曲数少)和形成期(感染后到出现的时间)有差异的两种孢子(Iwano & Ishihara,1991)。本研究确认感染家蚕幼虫的N.bombycis在寄主体内也和昆虫培养细胞内一样,形成异型的两种孢子,特别对短极丝型孢子的生物学及在养蚕方面的意义进行了考察。     

朱砂蛾Tyria jacobaeae中的两种寄生性微孢子虫：Nosema tyriae n．sp．和Nosema sp．

在朱砂蛾幼虫(Tyria jacobaeae)63%的群体中发现了Nosema tyriae.sp.感染发生于肠壁、丝腺、脂肪体中,可以说感染面相当广但致病性很低。裂殖体增殖(merogony)和孢子生殖(sporogony)在偶核期(diplokaryotic stages)进行双分裂(bina-ry fission)方式繁殖。新鲜孢子延伸到前末端,大小为4.7×2.0um。将产孢体(sporont)表面和寄主细胞的细胞质联系在一起的20nm长的微管的超微结构引起了人们的兴趣,在孢子中,一种圆形极囊,包含了纵向紧密排列的细胞膜和横向松散排列的细胞膜,以及单列上有10.5-14圈极丝。N.tyriae和来源于家蚕(Bombyx mori)的桑蚕微孢子虫(Nosema bombycis)的16SrRNA基因只有六个核苷酸不同。N.tyriae对家蚕微孢子虫的单克隆抗体表现出中度的阳性反应。N.tyriae对家蚕具有感染性但毒性比N.bombycis要低。但是用N.bombycis的特异性引物扩增N.tyr-iae DNA却未得到扩增产物,两者的孢子形状也各不相同。另一种仅对其中一条家蚕幼虫具有轻微感染性的双核微孢子虫Nosema sp.具有更短的发育期,孢子大小为3.8×2.0um(固定)。超微结构上也极为不同,它的裂殖体(meront)和产孢体(sporont)中的细胞质囊泡均有大量致密的膜。从该微孢子虫感染的幼虫组织中可以观察到两种微管排列:一种具有约15圈极丝成不规则的成簇排列,或是以12圈极丝排成一列。由于该幼虫是和被N.tyriae感染的幼虫一起保存的,所以这两种孢子很可能是来自双重感染。     

家蚕微孢子虫保存株DNA的特异性扩增

尝试研制成了一种微孢子虫检测的新系统。以家要微孢子虫小亚单位rRNA基因的推导假基因及保守序列为引物,对家蚕微孢子虫(Nosemabombycis.Nb)保存株的DNA进行了特异的PCR扩增。检测了六株微孢子虫:三株为家蚕微孢子虫株系,即保存株SES—NU、NIS—001和分离自草地贪夜蛾(Spodoptera depravata)的株系Sd—NU—IW8401;及三个种:Nosema Sp.株NIS—Mll,变形孢子虫属的分离株Vainmorpha Sp.NIS—M12和具褶孢子虫属分离株Pleistoophora Sp.PI—NU,以此六株微孢子虫的DNA为模板进行PCR扩增,仅家蚕微孢子虫(Nb)的保存株SES—NU、NIS—001获得扩增产物。     

家蚕病原性微孢子虫SCM_7（Endoreticulatus．sp）的分离和研究

从四川省种茧育家蚕分离出的一种小型微孢子虫ＳＣＭ７，仅感染寄生蚕的中肠上皮组织；孢子卵圆形，大小２．２６±０．２１×１．１９±０．１８μｍ，孢子具单核，极丝排列７─９圈，各阶段的发育均发生在由寄主细胞内质网膜所形成的寄生囊内。其它特点也与Ｅｎｄｏｒｅｔｉｃｕｌａｔｕｓ属相符，应分类于Ｅｎｄｏｒｅｔｉｃｕｌａｔｕｓ属。该属微孢子虫在家蚕体内尚属首次发现。     

高寒地区禾本科牧草生产力适应性评价

在玉树县巴塘乡铁力角村冬春退化草地上,开展多年生禾本科牧草引种试验,共引种7属15个品种。2年的试验结果表明,15个品种在巴塘地区均能正常越冬并完成生育期。牧草产量和种籽产量比较表明,其中同德老芒麦(E. sibiricus L.'Tongde')第2年干草产量最高为10030.8 kg·hm^-2,同德短芒披碱草(E. Breviaristatus Keng f.'Tongde')草籽产量最高,达到2986.7 kg·hm^-2。所有牧草品种均能完成生育期,并且第2年株高、盖度和产草量均高于第1年,适应玉树地区的生态环境,生产力均较好,适宜在玉树地区推广。     

草地早熟禾转CMO-BADH双基因和转CMO基因耐盐性分析

通过对非转基因草地早熟禾(Poa pratensis L.)、转CMO-BADH双基因草地早熟禾、转CMO基因草地早熟禾进行不同浓度的NaCl胁迫试验,测定其细胞膜的透性,丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性、氧化氢酶(CAT)活性,评定各株系耐盐能力的强弱,同时还验证并比较了CMO-BADH双基因和CMO基因的耐盐性功能,为耐盐新品种的选育提供了理论依据。结果表明:所有株系的相对电导率、MDA含量均随盐浓度增加而增大,在NaCl胁迫下相对电导率和MDA含量的大小顺序为:非转基因株系>转CMO基因株系>转CMO-BADH双基因株系;在NaCl的胁迫下,各个株系的SOD活性、POD活性、CAT活性都有明显的提高,此3种指标的大小顺序为:转CMO-BADH双基因株系>转CMO基因株系>非转基因株系;综合考虑各个指标,各株系耐盐性强弱顺序为:转CMO-BADH双基因株系>转CMO基因株系>非转基因株系。     

不同饲料对中华倒刺鲤仔稚鱼消化酶活力的影响

采用生物化学的方法研究中华倒刺鲃仔稚鱼消化酶的个体发生和相互之间的异同.得到以下结果从胰蛋白酶活力来看.3个试验组之间具有显著性差异(P＜0.05),在试验后期,活饵组的胰蛋白酶活力大部分时间都大于饲料组.从淀粉酶活力来看,3个试验组都具有稳定上升的共同趋势,不过3个试验组之间仍旧具有显著性差异(P＜0.05).从脂肪酶活力来看,试验期间的变化规律都十分剧烈,其活力交替上升,3个试验组之间具有极显著性差异(P＜0.01).由此可见,不同饲料对于中华倒刺鳃仔鱼消化酶活力的影响十分巨大.     

Degenerative mucinotic mural folliculitis in cats

A novel form of mural folliculitis is described in seven cats. Clinically, all cats exhibited generalized alopecia with scaling or crusting that was more pronounced over the head, neck, and shoulders. The face and muzzle of all cats was unusually thickened. Six of seven cats were progressively lethargic but did not demonstrate any other consistent systemic abnormalities. Histologically, there was severe mixed inflammation of the wall of the follicular isthmus in all cats, accompanied by some follicular destruction in five cats. Sebaceous glands were not affected. All cats had variable, but often striking, follicular mucin deposition, as well as epidermal hyperkeratosis and crusting. The cause of the severe mural folliculitis was not identified, and all cats responded poorly to immunomodulating therapy. Follicular mucinosis may be a nonspecific finding, likely reflective of the follicular lymphocytic milieu, and does not always herald follicular lymphoma.     

西藏不同饲草全混合日粮发酵品质和有氧稳定性的研究

为了充分利用西藏的农作物秸秆和饲草资源,本试验在当地奶牛场采用的以全株玉米和青稞秸秆为主要粗饲料的TMR配方基础上,用苜蓿、全株燕麦和小麦秸秆按不同比例替代全株玉米。青贮发酵45 d后评定各组发酵品质、营养特性和有氧稳定性。结果表明,发酵45 d后,TMR-2、TMR-3和TMR-4组pH值显著高于TMR-1(P<0.05),乳酸含量均低于TMR-1,但各替代组仅检测到微量丙酸和丁酸含量,且氨态氮/总氮较低,表明各组发酵品质均良好。发酵后各组水溶性碳水化合物含量均有大幅度下降,除TMR-1组外,其他3组干物质含量基本不变,粗蛋白、中性洗涤纤维、酸性洗涤纤维和粗脂肪等营养物质含量不同程度增加,其中TMR-2和 TMR-3组粗蛋白含量高于15%,能够满足改良奶牛的营养需要。在有氧暴露过程中,各组乳酸、乙酸和水溶性碳水化合物含量均出现不同程度的下降,丙酸和丁酸含量在有氧暴露第3天均有升高,之后逐渐下降。在有氧暴露9 d中,pH值均呈上升趋势,其中TMR-3组pH值上升幅度最小,其有氧稳定性最好。综上所述,虽然TMR-3发酵品质略低于TMR-1组,但其有氧稳定性有所提高,因此,TMR-3组可应用于生产实际中。     

鸡EX-FABP基因PCR-SSCP分析

以AA白鸡、广西鸡、E1矮小鸡品系为试验材料,采用PCR-SSCP技术对鸡细胞外脂肪酸结合蛋白基因(EX-FABP)的2个片段进行PCR扩增,分析了EX-FABP基因在3个鸡品系的多态性。结果表明:EX-FABP基因在2对引物扩增片段中均存在PCR-SSCP多态性。对于引物1扩增片段,3个鸡品系均检测到BB基因型,AA基因型只出现在AA白鸡中;而且B等位基因频率在3个鸡品系中均明显高于A等位基因频率。对于引物2扩增片段,3个鸡品系均检测到HH和Hh基因型,hh基因型只出现在E1矮小鸡中;H等位基因频率在3个品系中均明显高于h等位基因频率。引物1和引物2的多态性片段测序分析表明,EX-FABP基因第641位点和645位点分别发生了单碱基的转换(G-A和T-C),第3264位点发生了转换(T-C)。     

Immunolocalization of epidermal growth factor (EGF) and EGF receptors in the porcine upper gastrointestinal tract.

The factors controlling growth and maturation in the porcine gastrointestinal tract are not well understood. Epidermal growth factor (EGF) is a polypeptide that has been implicated in the control of gastrointestinal tract growth, maturation, and protection in other species. Immunoreactive EGF (IR-EGF) and EGF receptors (EGF-R) were histochemically identified in formalin-fixed tissues of the upper digestive tract of 1-, 3-, 7-, 14-, 21-, and 28-day-old pigs. The ductal epithelium consistently contained IR-EGF in the parotid salivary gland of pigs of all ages and in the mandibular salivary gland in pigs greater than or equal to 7 days old. Immunoreactive EGF was detected in the mucosal epithelium of the esophagus and nonglandular portion of the stomach, and in the pancreas and liver in all pigs. Gastric gland IR-EGF was inconsistently detected in pigs less than 14 days old and was consistently observed in all older pigs. Enterocyte EGF immunoreactivity was usually weak and was variably detected in the duodenum of pigs less than or equal to 7 days old and in the jejunum of pigs less than or equal to 14 days old, but was consistently observed in older pigs. Ileal immunoreactivity was erratic. Immunoreactive EGF-R were identified in the esophageal epithelium of all pigs, and in the nonglandular gastric and glandular gastric mucosa of all pigs, except for two 7-day-old pigs and one 7-day-old pig, respectively. Immunoreactive EGF-R were detected in the duodenal, jejunal, and ileal enterocytes of pigs of all ages examined.     

不同秋眠类型苜蓿品种生产性能与持久性分析

通过对58份不同秋眠类型苜蓿品种再生性、持久性和产草量在呼和浩特的测定分析,结果表明:1)在全部参试材料与秋眠类型组中,苜蓿秋眠级和再生性正相关并达到极显著、显著水平;而在半秋眠类型组、非秋眠类型组中,苜蓿的再生性和秋眠级相关性很小。2)在全部参试材料、半秋眠类型组、非秋眠类型组中,首蓿的持久性与秋眠级负相关,但是在秋眠类型组中,首蓿秋眠级和持久性相关性很小。3)在全部参试材料、半秋眠类型组、非秋眠类型组中,苜蓿的产草量和秋眠级负相关;在秋眠类型组中,苜蓿的产草量和秋眠级正相关但不显著。     

EVALUATION OF PROCEDURES FOR THE DIAGNOSIS OF BRUCELLA OVIS INFECTION IN RAMS

Procedures for the diagnosis of Br. ovis infection in rams were evaluated by examining 10 rams artificially infected by preputial inoculation. Observations were undertaken at weekly intervals for 1 year to follow changes in clinical, bacteriological and serological findings. Clinical lesions were detected in 1 ram 3 weeks after inoculation and in all rams by 8 weeks; lesions were undetectable in 3 rams at the completion of the trial. The presence of inflammatory cells in semen samples was the earliest indication of infection being demonstrated in 2 rams at 2 weeks and in all rams by 8 weeks; subsequently 86% of samples were positive. Br. ovis was detected in semen smears from 3 rams at 4 weeks but only once in all rams (at 27 weeks); overall 52% of semen smears were positive from 4 weeks onwards. Br. ovis was cultured from semen of 5 rams after 4 weeks and from all rams at 5 weeks; thereafter 97% of samples were positive. All rams developed significant titres to the CFT between 2 and 9 weeks; thereafter the CFT was a reliable indication of infection in 6 rams, highly suggestive in 3 and unreliable in one. By 8–10 weeks all rams developed significant titres to the IHA which were then maintained in all rams for the remainder of the trial.     

Response of chickens with genetically different hemoglobin types to infection with Eimeria tenella and mechanical bleeding.

The hematological and body-weight responses of chickens with mutant and normal hemoglobin types were compared under conditions of hematopoietic stress from coccidiosis infection as well as blood loss from mechanical bleeding. A dose of 100,000 Eimeria tenella oocysts caused a highly significant decrease (about 30%) in the packed cell volume, resulting in a severe anemia in all hemoglobin types. Hematological response to the anemia produced by the coccidiosis was similar for all three hemoglobin types. Although body weights and gain of all hemoglobin types infected with coccidiosis were significantly depressed, there was no differential response between the mutant and normal hemoglobin types. Mechanical bleeding resulted in a hypochromic normocytic anemia in all hemoglobin types. Ability to compenstate for blood loss was similar for the mutant and normal hemoglobin types.