鸡毒支原体敏感株与耐药株超微结构的比较

对临床分离和实验室药物压力下筛选的鸡毒支原体耐药株与敏感株进行超微结构的观察和比较.结果显示鸡毒支原体敏感株呈多形性,柔软且有较大的变形性,可清晰观察到细胞膜分为外、中、内三层膜,并可观察到裂殖繁殖方式;有的支原体在繁殖时先在极端产生泡状突起,形成不均等分裂.在恩诺沙星药物压力下敏感株产生耐药性后其外膜显著增厚,导致支原体的多形性减弱或消失,呈现出较为一致的圆形;细胞膜内层周围存在排列整齐、结构紧密、类似微管样的结构,胞内电子密度明显升高.临床分离的耐药株超微结构观察结果与实验室条件下筛选的耐药株一致:凡是超微结构发生变化的,均存在耐药表型,而且高水平耐药株的超微结构变化最为突出.研究结果表明耐药性的产生可导致鸡毒支原体超微结构明显的改变,并可能引起抗原性变异.     

鸡毒支原体病及其防控

鸡毒支原体感染,又称鸡败血支原体感染或鸡慢性呼吸道病(CRD),是由鸡毒支原体引起的鸡和火鸡的慢性呼吸道传染病。病鸡主要表现为呼吸道症状,如气管炎、气囊炎等,以咳嗽、气喘、流鼻液和呼吸锣音为特征。该病病程缓慢、病程长,成年鸡多     

抗菌药物联合使用对耐达氟沙星鸡毒支原体的体外抑制作用研究

为探究抗菌药物联合使用对耐达氟沙星鸡毒支原体的体外抑制效果,选择鸡毒支原体标准株S6和对达氟沙星MIC升高程度不同的突变株M1、M2和M4,分别测定泰乐菌素、大观霉素、林可霉素、泰妙菌素、替米考星和多西环素种对4株鸡毒支原体的最低抑菌浓度,并使用达氟沙星分别联合上述6种抗菌药物进行体外抑菌试验.结果显示:除达氟沙星外,...     

壮观霉素对体外鸡毒支原体的抑菌试验和对鸡体内鸡毒支原体病的治疗试验

1　材料与方法1 .1　试验药物　壮观霉素为水溶性粉剂 ,批号940 71 2 ,由中国医学科学院医药生物技术研究所北抗药厂生产。对照药物壮观霉素 ,水溶性粉剂 ,由比利时生产 ,批号 573KS,由中国医学科学院医药生物技术研究所提供。1 .2　鸡毒支原体菌种　 R株、S6株 ,均由中国兽医药品监察所保存。1 .3　试验鸡　 9日龄北京白鸡 ,试验前经血清学检测 ,该鸡为鸡毒支原体血清学阴性反应。1 .4　鸡毒支原体血清平板凝集抗原　由中国兽医药品监察所生产 ,批号为 930 1。1 .5　抑菌试验　试验前将试验药壮观霉素用蒸馏水配成一定浓度的溶液过滤除菌…     

甲磺酸培氟沙星对体外鸡毒支原体的抑菌试验和对鸡毒支原体病的治疗 …

利用甲磺酸培氟沙星水溶性粉剂对鸡毒支原体分别进行体内外的抑菌治疗试验。体外抑菌试验结果表明：该药物能有效地抑制试管中鸡毒支原体的生长繁殖。体内试验结果表明：当饮水中药物浓度达到１００ｍｇ／Ｌ时,能有效地防止由于鸡毒支原体引起的死亡;当饮水中药物浓度达到５０ｍｇ／Ｌ以上时,能明显地减轻以至部分消除鸡毒支原体强毒引起的气囊损伤;和感染不对照组相比,该药物能提高鸡体增重;用甲磺酸培氟沙星治疗,能抑制感染     

甲磺酸培氟沙星对体外鸡毒支原体的抑菌试验和对鸡毒支原体病的治疗试验

利用甲磺酸培氟沙星水溶性粉剂对鸡毒支原体分别进行体内外的抑菌治疗试验。体外抑菌试验结果表明：该药物能有效地抑制试管中鸡毒支原体的生长繁殖。体内试验结果表明：当饮水中药物浓度达到１００ｍｇ／Ｌ时,能有效地防止由于鸡毒支原体引起的死亡;当饮水中药物浓度达到５０ｍｇ／Ｌ以上时,能明显地减轻以至部分消除鸡毒支原体强毒引起的气囊损伤;和感染不治疗对照组相比,该药物能提高鸡体增重;用甲磺酸培氟沙星治疗,能抑制感染鸡体内的鸡毒支原体,从而使部分鸡体内检测不到鸡毒支原体抗体。     

四川地区鸡毒支原体的分离鉴定及血清学定型

在成都、重庆等１０个市、地、县,从患有慢性呼吸道疾病的鸡场采集１０８份样品,分离的２７个菌株,经Ｌ型细菌检验,理化特性鉴定,符合支原体的生物学特性。通过血清学鉴定（生长抑制试验）,证实分离的支原体与鸡毒支原体国际代表株Ｓ６为同一血清型。人工复制试验结果,表明分离的鸡毒支原体ＳＭＧ－１和ＳＭＧ－２具有高致病性,感染鸡出现典型的慢性呼吸道病症状     

白虎定喘口服液的质量检测

为了建立白虎定喘口服液的质量检测标准,采用薄层色谱法(TLC)对处方中金银花、黄芩、知母、板蓝根进行鉴别;采用高效液相色谱法(HPLC)对黄芩中有效成分黄芩苷进行含量测定。结果表明,薄层色谱斑点清晰,分离度好,特异性强;在0.103mg/mL~0.824mg/mL范围内黄芩苷浓度与峰面积呈良好的线性关系,阴性对照无干扰,加样回收率平均为97.98%,精密度试验RSD为0.61%。说明建立的检测方法简便、准确、专属性强、重复性好,可对白虎定喘口服液的质量进行有效检测。     

白虎定喘口服液药效研究

为验证新兽药白虎定喘口服液的药效作用,进行了鸡传染性支气管炎病毒（infectious bronchitis virus,IBV）鸡胚增殖抑制试验、气管纤毛运动试验及SPF雏鸡人工感染IBV疗效试验。将试验药液与等量IBV液混合作用后,接种11日龄SPF鸡胚,37℃孵育168 h,取存活鸡胚肾脏进行IBV的RT-PCR检测,结果表明,药物组168 h鸡胚存活率为80%,阳性对照组为20%,两组间差异显著（P<0.05）;试验药物组RT-PCR检测无目的条带出现,而阳性对照组有目的条带出现。40日龄SPF鸡连续用药3 d后,气管内注入0.02 mL墨汁,测量1 min内墨汁移动的距离,结果表明药物组墨汁移动距离极显著大于生理盐水对照组（P<0.01）。17日龄SPF鸡用IBV-M41滴鼻,0.3 mL/只,24 h后开始灌服药液,1 mL/kg,连用5 d,结果表明,试验药物组与攻毒对照组在临床症状计分、痊愈率上均存在显著差异（P<0.05）;试验组气管黏膜组织切片未见明显病变,而阳性对照组有明显病理变化。以上试验结果表明,白虎定喘口服液对IBV具有较好的体外抑制作用,可显著促进气管纤毛的运动,对鸡传染性支气管炎（IB）具有较好的临床防制效果。     

Study on Efficacy of Baihu Dingchuan Oral Liquid

In order to verify the effect of Baihu Dingchuan oral liquid of new veterinary drugs,the effects of infectious bronchitis virus (IBV) on the proliferation inhibition test, tracheal cilia movement test and SPF chickens infected with IBV were tested.The test solution was mixed with an equal amount of IBV solution,inoculation of 11-day-old SPF chicken embryo,incubation at 37℃ for 168 h.RT-PCR was performed to detect IBV in kidney of live chick embryo.The results showed that the survival rate of 168 h chicken embryo was 80% in the drug group,but the positive control group was 20%, and there was significant difference between two groups (P<0.05). In the test drug group, no band was detected by RT-PCR, but the target band appeared in the positive control group.After 40 days old SPF chickens were administered continuously for 3 days, 0.02 mL of ink was injected into the trachea. The distance of ink movement in 1 min was measured. The results showed that the migration distance of ink was extremely significantly higher than that of saline control group (P<0.01).In the 17-day-old SPF chickens, IBV-M41 was infused into the nasal mucosa with 0.3 mL. After 24 h, 1 mL/kg was administered for 5 days. The results showed that there were significant differences in clinical symptom score and cure rate between the test drug group and the control group (P<0.05). There was no significant pathological changes in the tracheal mucosa of the experimental group, but the pathological changes in the positive control group were significant. The above results showed that Baihu Dingchuan oral liquid had good inhibitory effect on IBV in vitro, which could promote the movement of tracheal cilia, and had good clinical control effect on infectious bronchitis.     

德国牧羊犬精子超微结构及超低温冷冻对精子超微结构的影响

为了研究超低温冷冻前后德国牧羊犬精子超微结构的变化,采用扫描电镜和透射电镜对冷冻前后德国牧羊犬精子超微结构变化进行研究,结果显示在扫描电镜下,鲜精整体呈“蝌蚪状”,头部呈扁卵圆形.在透射电镜下精子纵切面头部呈“楔形”,头部细胞膜由外向内分别由质膜、顶体外膜、项体内膜和核膜组成.精子尾部中段横切面由外向内可见线粒体鞘膜、9束外周致密纤维,9对轴丝和2根中央微管.精子头长约为5.40 μm,头宽3.26 μm,颈长1.25 μm,颈宽0.55 μm,尾部中段长11.34 μm,中段直径0.87 μm,尾部长55.70 μm,线粒体螺旋数为44旋.鲜精和冷冻前精子头部、颈部和尾部形态变化差异均不显著(P＞0.05).超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数极显著高于鲜精和冻前精子数(P＜0.01).冻后顶体发生明显形态变化的精子占54.21％,高于发生颈部形态变化(10.61％)和尾部形态变化(28.83％)的精子数.结果表明鲜精和冻前精子形态变化不显著,而超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数显著高于鲜精和冻前精子数.     

Thrombocyte morphology and morphometric observations in two vulture species

Background: The important role of thrombocytes in hemostasis is well documented, but little information is available on the thrombocyte morphology of avian species, including vultures. Objective: The objective of this study was to describe and compare the morphology and morphometric parameters of thrombocytes in 2 vulture species. Methods: Blood samples were collected into tubes containing acid‐citrate‐dextrose from 5 Cape vultures (Gyps coprotheres) and 6 white‐backed vultures (Gyps africanus) at the De Wildt Breeding Center, Northwest Province, South Africa. Wright's‐stained blood smears were examined by light microscopy. Samples were processed and examined by scanning and transmission electron microscopy (TEM) using standard techniques. Morphometric parameters (perimeter, area, minimum and maximum diameter, and aspect ratio) were measured on 140 thrombocytes using imaging software. Results: Thrombocytes were predominantly oval to elliptical, with few pseudopodia. The nucleus was the most prominent feature of the cells. Large vacuoles were visible in the cytoplasm by both light and TEM. Ultrastructurally, microtubules and dense bodies were seen in most cells. Other cytoplasmic organelles seen by electron microscopy included mitochondria, endoplasmic reticulum, a surface connecting canalicular system, Golgi complex, lipid droplets, and glycogen. Thrombocytes of Cape vultures had a significantly (P=.005) higher aspect ratio compared with white‐backed vultures. Thrombocyte estimates in blood smears were similar in both species, with a combined mean of 31.6 × 10⁹/L. Conclusion: The morphologic features of thrombocytes in southern African vultures are similar in most ways to those of other avian species. Although thrombocytes in white‐backed vultures were slightly more spherical than those of Cape vultures, no other significant differences were found between the 2 species.     

马度米星铵对成肌细胞转录组和超微结构的影响

探究马度米星铵对成肌细胞基因表达和超微结构的影响。马度米星铵处理C2C12细胞后,于细胞实时分析仪中记录细胞生长情况;利用Illumina HiSeq平台进行转录组测序,筛选差异表达基因并进行基因本体富集分析;显微镜下观察细胞空泡情况并拍照;透射电镜下观察拍照细胞超微结构。结果表明,马度米星铵抑制C2C12细胞生长,呈浓度和时间相关性;1 μM马度米星铵处理细胞8 h后,差异表达基因共2450个,其中下调基因为1411个,上调基因1039个;基因本体分析发现,差异表达基因显著富集到空泡、DNA修复、转移酶活性正调控等;细胞内空泡显著增加;1 μM马度米星铵处理细胞24 h后,细胞器明显肿胀、线粒体嵴结构髓样变、粗面内质网表面附着核糖体脱颗粒。由此可知,马度米星铵体外可诱导C2C12细胞空泡化,破坏细胞器结构,抑制细胞生长,从而导致其骨骼肌成肌细胞毒性作用,干预细胞空泡化是预防和治疗马度米星铵中毒的重要途径。     

胰岛素对雏鸡心肌发育及其超微结构的影响

选取16只体重相近的健康雏鸡,随机分为两组。试验组每天定时腹腔注射生理剂量胰岛素,对照组腹腔注射等剂量的生理盐水。连续注射10d后称重,剖检取样并制片,通过分析心体比和透射电镜观察雏鸡心肌的超微结构变化,探讨胰岛素对雏鸡心肌发育及其超微结构的影响。结果表明,与对照组相比,试验组雏鸡的体重、心脏重和心体比没有显著变化,但其心肌的线粒体数量和横截面积明显增加。提示生理剂量胰岛素有促进心肌细胞代谢的作用。     

Hematology,morphology, cytochemical staining,and ultrastuctural characteristics of blood cells in king cobras (Ophiophagus hannah)

Background — King cobras (Ophiophagus hannah) have been captive‐bred at Queen Saovabha Memorial Institute since 1996 to supply venom for antivenom production. Hematologic tests would be useful for evaluating the health of the snakes, however, basic hematologic data and morphology have not been described for this species. Objective — The purpose of this study was to determine basic hematologic values and evaluate light microscopic, cytochemical, and electron microscopic characteristics of king cobra blood cells. Methods — Blood samples from 13 wild‐caught and 15 captive‐bred king cobras were collected into EDTA from the ventral caudal vein. A CBC was done using standard methods. Significant differences between groups were determined using t‐tests. Cytochemical stains (periodic acid‐Schiff [PAS], Sudan black B [SBB], a‐naphthyl acetate esterase [ANAE], acid phosphatase [AcP], and p‐glucuronidase [p‐glu]), and scanning and transmission electron microscopy were done using standard techniques. Results — Eighteen snakes (64.3%) were positive for Hepatozoon infection. Hepatozoon organisms were detected nearly twice as frequently in wild‐caught (11/13) as in captive‐bred (7/15) snakes.Total WBC, azurophil, and lymphocyte counts were higher and fib‐rinogen concentration was lower in Hepatozoon‐positive snakes. Captive‐bred snakes had higher RBC values, lower azurophil, het‐erophil and punctate reticulocyte percentages, and higher lymphocyte numbers compared with wild‐caught snakes. Lymphocytes were the most commonly observed WBCs, and stained positive with PAS, ANAE, AcP, and (3–glu. Azurophil granules stained positive with SBB, PAS, and ANAE. Heterophils were the largest WBCs; their granules stained with SBB, ANAE, and (3–glu. Basophil granules stained with PAS, SBB, ANAE, and (3–glu. Thrombocytes were strongly positive with PAS. Transmission electron microscopic examination revealed organelles within all WBCs except eosinophils and revealed the gamonts of Hepatozoon sp in RBCs and azurophils. Conclusions — These results provide comparative hematologic data and a guide for identification of blood cells in wild‐caught and captive‐bred king cobra snakes. Hepatozoon infection was relatively common, but was not associated with severe hematologic abnormalities.     

比格犬甲状腺显微与超微结构的观察

应用光镜和透射电镜技术，观察了3～6月龄比格犬甲状腺的显微和超微结构。结果表明，比格犬甲状腺实质由滤泡和滤泡旁细胞构成。滤泡呈圆形或椭圆形，直径20．22～220．00μm，平均90.80μm；由单层立方上皮细胞围成，细胞高度3．04～7.11μm，平均5.18μm，电镜下可见功能状态不同的两型滤泡上皮细胞；滤泡旁细胞很多，直径4．40～8．82μm，平均6．23μm，位于滤泡之间或镶嵌于滤泡上皮细胞之间，也可聚集在一起形成滤泡样结构，胞质内含有大量的分泌颗粒，电镜下也可见两种类型的滤泡旁细胞。     

不同诱变处理对苜蓿叶片细胞显微和超微结构的影响

研究⁶⁰Co-γ射线、紫外线、零磁空间和EMS诱变处理下苜蓿SP1代叶片细胞显微、超微结构的变化,筛选最优诱变处理,选育适合高寒地区种植的苜蓿新品系或品种。以公农1号、Wega7F、WL319HQ、敖汉4个苜蓿品种试验材料,设置⁶⁰Co-γ射线150、300、450 Gy射线强度,紫外线30、60、90 min照射时间,甲基磺酸乙酯(EMS) 0.1%、0.2%、0.4% (v/v) 溶液浓度,零磁空间处理180 d;测定苜蓿叶片细胞显微和超微结构。不同诱变处理后4个苜蓿品种叶片厚度均不同程度增加,其中公农1号EMS 0.4% (v/v)浓度下,叶片厚度最大,为446.16 μm,增厚幅度达到25.86%,同时诱变降低了叶脉突起程度;公农1号⁶⁰CO-γ辐射和紫外线处理细胞结构疏松度均高于对照,诱变后Wega7F细胞结构紧密度升高;EMS处理下海绵组织厚度均降低;低剂量诱变处理下[150 Gy ⁶⁰Co-γ,30 min紫外线,0.1% (v/v) EMS],细胞叶绿体出现变形、基粒片层松散、类囊体解体、脂质球增多等现象,高剂量处理[450 Gy ⁶⁰Co-γ,90 min紫外线,0.4% (v/v) EMS]和零磁空间处理下,叶绿体膜会有部分模糊或解体,基粒片层膨胀或模糊、基粒垛叠程度变化随诱变处理种类增加或降低。4种苜蓿品种的叶片厚度诱变后叶片厚度均增加。在一定梯度范围内,⁶⁰CO-γ射线和EMS诱变效率较高,突变体材料丰富,操作简单,适宜诱变育种的实际操作;150 Gy, ⁶⁰Co-γ射线、60 min紫外线、0.4%浓度EMS处理为苜蓿正向变异处理,有利于筛选有益诱变突变体。