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1.
为探究支原体液体培养基和固体培养基质控菌猪鼻支原体生长规律,以活菌浓度,菌液pH值为指标,比较了不同代次与不同传代方式猪鼻支原体生长情况。结果显示:3%与10%浓度传代,0~48 h内,活菌浓度持续上升,菌液pH值持续下降,3%浓度传代的支原体生长较均匀,更利于菌种生长;1~5代支原体活菌浓度无显著差异;通过0.3 mL菌液加9.7 mL培养基传代,20组菌液平均浓度为(24.4±5.7)×10~8CFU/mL,通过0.9 mL菌液加29.1 mL培养基传代,菌液平均浓度为(7.2±2.1)×10~8CFU/mL,两种浓度差异极显著,前种传代方式更有利于支原体生长。本次试验为支原体液体和固体培养基的质控工作标准化提供参考。 相似文献
2.
对发酵黄芪样品中分离的1株产纤维素酶解淀粉芽胞杆菌的生物学特性进行了检测,试验结果表明,菌株在10℃~60℃环境均能生长,最适生长温度为35℃~40℃。pH5.0~8.0培养基均适宜菌株生长,其中最适pH为6.0。在37℃环境200r/min振荡培养的生长曲线在0~4h为延迟期,4h~12h为对数期,12h~24h为稳定期,24h以后为衰亡期。细菌37℃培养32h开始产生芽胞,在培养72h时芽胞形成率达到80.67%。菌株对多数抗菌药物敏感,且安全性良好。该菌株适应性强,生长条件较为宽松,这为其开发应用提供了依据。 相似文献
3.
1976年从猪喘气病的病肺中分离到两株支原体培养物。在液体培养基中生长迅速,菌体以姬姆萨染色呈紫色小点状多形态。固体培养基上生长小圆球状菌落,中间有一小突起。鸡胚卵黄囊内接种,7至10天后,出现心包炎。实验感染三周龄仔猪,腹腔内接种能诱发典型的多发性浆膜炎。作者曾暂定为猪鼻支原体。后经上海畜牧兽医研究所进行生长抑制试验鉴定,N_3和 N_5株与猪鼻支原体国际标准首株 BTS_7栋为同种(10)。所以 N_3和 N_5株应为我国首次分离的猪鼻支原体菌株,并证实我国猪喘气病病猪的病肺中也存在着猪鼻支原体。 相似文献
4.
为筛选出山羊支原体山羊肺炎亚种的适宜培养基,将山羊支原体山羊肺炎亚种分离株M1601分别接种Thiaucourt肉汤、MEM-KM2和TSA 3种不同的培养基,测定其在3种不同培养基中生长滴度、生长速度,并测定了M1601在最适培养基中在不同培养阶段的生长滴度。结果表明,添加2g/L丙酮酸钠和150mL/L马血清的Thiaucourt肉汤培养基最适宜山羊支原体山羊肺炎亚种的生长,生长滴度可达109 CCU/mL,在培养6h后开始进入对数生长期,培养60h后进入稳定期,培养72h时后进入衰亡期。此试验结果为山羊支原体山羊肺炎亚种培养特性研究提供了参考数据。 相似文献
5.
为了比较《中国药典》已收录但相当耗时的液体培养法和未收录但相对快速的荧光定量PCR(qPCR)法对支原体检测的灵敏度，试验分别在猪鼻支原体生长的对数期（48 h）、稳定期（96 h）和衰亡期（144 h）取样，将样品10倍系列稀释后，使用液体培养法和qPCR法对各个稀释度进行检测（两种方法的样品加入体积分别为0.5 mL和5μL），以比较二者的灵敏度。结果显示：在存活支原体占比高的对数期和稳定期，液体培养法比qPCR法的检测灵敏度高约100倍；而在死亡支原体占比大幅增加的衰亡期，结果则相反，qPCR法反而比液体培养法的检测灵敏度高约1 000倍。当把对数期样品高速离心浓缩约100倍后再用qPCR法检测，其检测灵敏度比浓缩之前提高了约100倍。上述结果表明：影响液体培养法和qPCR法相对灵敏度的主要因素是各自体系中不同的样品加入体积和样品中存活支原体的比例，而由于前一因素导致的两种方法灵敏度的差异，可以通过高速离心浓缩的方法缩小，使两种方法的相对灵敏度基本相同。 相似文献
6.
为研究适用于水貂的专用菌制剂,试验采集健康成年母貂的新鲜肠粘膜和粪便,用改良MRS培养基,对肠道与粪便中的微生物进行分离培养,筛选优势菌株,观察其菌落形态,并进行生化试验和16SrRNA分子生物学鉴定,并对其生长情况和安全性等生物特性进行了研究。结果表明,优势菌株(RSJ)为乳酸杆菌;生长6h进入对数期,前18h菌株繁殖较快,pH急速下降;18~36h,菌株维持在稳定期;pH 2.0和3.0时菌株不生长,pH 5.0和6.0时生长良好;对青霉素和环丙沙星的耐药性较强,对其余的抗生素则表现为敏感;菌株RSJ对金黄色葡萄球菌、大肠杆菌均有抑菌作用。安全性试验显示,RSJ对小鼠无毒性。RSJ菌株具备了益生菌的基本条件,为进一步研究与开发微生态制剂奠定了基础。 相似文献
7.
支原体检验用培养基的研究 总被引：2，自引：2，他引：0
水质和血清对比试验结果表明：以水质电导率低于0．5μs／cm、猪血清总蛋白不低于49g／L、胆固醇不高于1．66mmol／L为标准制备的改良Frey培养基和支原体培养基活菌滴度均达10^9CCU／mL。对滑液支原体GX11-T株及猪鼻支原体BTS-7株1—5代对数生长期培养物检测,均达10^9CCU／mL。改良Frey培养基与支原体培养基经反复冻融8次;或改良Frey培养基经37℃保存24h;支原体培养基37℃保存8h均不影响灵敏度。改良Frey培养基在鸡胚成纤维细胞及鸡胚尿囊液中最小检测量分别为4CCU／mL和2CCU／mL;支原体培养基在BHK：,细胞中最小检测量为8CCU／mL。改良Frey培养基和支原体培养基4℃有效期为1个月,-20℃为12个月。 相似文献
8.
为了研究山羊支原体山羊肺炎亚种分离株M1601株在Thiaucourt肉汤培养基中的生长及繁殖特性。利用活菌计数方法对其在不同培养阶段的生长滴度及pH值进行了测定,并绘制了生长曲线。结果表明,传代稳定的M1601在Thiaucourt肉汤培养基中培养6～60h为对数生长期．pH值逐渐下降：培养60～72h进入稳定期．pH值降为6．27．培养72h进入衰亡期。这为山羊支原体山羊肺炎亚种培养特性研究和疫苗生产工艺研究提供了参考数据,以便在生产上更合理地应用其生长规律。 相似文献
9.
建立了仔猪副伤寒活疫苗生产用菌株生长曲线,获取菌株培养参数,从而实现合成培养基初步应用。将猪霍乱沙门氏菌(CVCC79500株)运用试管和三角瓶分别静置和振荡培养30 h,期间取4、8、12、18、24、30 h等6个点进行活菌计数并建立生长曲线,确定37℃静置培养18~24 h,37℃、200 r/min振荡培养8~24 h为培养稳定期。不同接菌量比较结果表明,37℃静置培养24 h,37℃、200 r/min振荡培养12 h均可达到菌体稳定期。在稳定期参数条件下,比较合成培养基与常规普通肉汤培养基对该菌的增殖效果,结果表明,37℃静置培养24 h,试管及三角瓶培养菌数分别为27×108~31×108、33×108~41×108CFU/m L;37℃、200 r/min振荡培养12 h,试管及三角瓶培养菌数分别为58×108~66×108、78×108~88×108CFU/m L。而同条件3批普通肉汤培养菌数均低于合成培养基,且批次间稳定性较合成培养基差。将合成培养基振荡培养12 h的菌液接种小鼠均10/10存活;免疫后攻毒家兔达4/5~5/5保护,与普通肉汤基本一致。合成培养基保存期试验表明,25℃避光保存28 d与当天配制的液体合成培养基,振荡培养菌数基本一致。获取的培养参数适合用于合成培养基的初步评价,研制的合成培养基培养菌数优于普通肉汤,且不影响菌体安全性及免疫原性,室温保存期可达28 d。 相似文献
10.
为探索在疫苗研制过程中牛支原体抗原收获时间及抗原定量替代方法,将牛支原体08M株接种于含10%马血清的Thiaucourt's培养基,在110 h内同时监测其颜色变化单位（color change units,CCU）、菌落形成单位（colony forming units,CFU）、菌体蛋白浓度和核酸含量的变化,绘制相应曲线。活菌计数结果（CCU和CFU）显示,牛支原体生长可分为明显的4期,10 h进入对数期,30 h进入稳定期,活菌数最高可达1.0×108 CCU/mL和7.7×107 CFU/mL,75 h进入衰亡期;蛋白浓度从15 h开始迅速增长,至35 h蛋白浓度最高,为72.06 μg/mL,此后维持在58.38~70.65 μg/mL;核酸含量从15 h开始增长,至25 h后Ct值维持在15.32~17.84。结果表明,牛支原体蛋白含量在稳定期初期与活菌数具有良好的相关性。因此,在牛支原体灭活疫苗生产中,稳定期初期是最佳抗原收获时间,可用蛋白浓度法代替活菌计数法进行抗原定量。 相似文献
11.
12.
This study attempted to determine whether one multiplex polymerase chain reaction (PCR) is an effective adjunct method for diagnosing Mycoplasma hyopneumoniae and Mycoplasma hyorhinis infection, and whether M. hyorhinis should be considered as an enzootic pneumonia or porcine respiratory disease complex pathogen in Taiwan. To our knowledge, this study is the first to isolate and identify M. hyorhinis as a porcine pathogen in Taiwan. A novel isolation method and a multiplex PCR test were applied to detect and isolate M. hyorhinis. The correlation of M. hyorhinis with swine pneumonia was also examined using a challenge test. Based on weight, 18 pigs were assigned to three groups and housed throughout the study in a specific-pathogen-free (SPF) facility and provided with aseptic feed and water. Groups 1 (n=6) and 2 (n=6) were challenged with 5mL M. hyorhinis culture via tracheal intubation on day 1. The M. hyorhinis strains ATIT-1, -3, and-7 were used to infect group 1 and the strain ATCC 27717 was used for group 2. Culture medium was replaced by phosphate-buffered saline in group 3 (n=6). All pigs were slaughtered on day 28, and their lungs were removed for examination of lesions. Of the six pigs in group 1 challenged with wild-type strains, two had typical mycoplasma pneumonia lesions. No gross lung lesions were observed in groups 2 and 3. Although further examination is necessary to confirm that wild-type strains can cause pneumonia, it appears that M. hyopneumoniae is no longer the only mycoplasma pathogen implicated in the diagnosis of swine enzootic pneumonia (SEP). 相似文献
13.
N. F. Friis 《Acta veterinaria Scandinavica》1976,17(3):343
In an examination of conjunctival samples from 40 piglets for mycoplasmas, 17 isolates were obtained. Eight could be identified as Mycoplasma hyorhinis, three as Mycoplasma flocculare, and one as Acholenlasma sp. Five strains were not readily identifiable, but together with two previously recovered strains they were found to represent a distinct serogroup. All seven strains were glucose and phosphatase positive. Incubation in a CO2-enriched atmosphere led to enhancement of the growth on solid medium. The serogroup was serologically related to M. hyorhinis, but not to a number of other glucose fermenting species of mycoplasma, and it may therefore be regarded as a new subspecies of M. hyorhinis. 相似文献
14.
The responses of the field strains and type strains of Mycoplasma hyorhinis are compared in epi-immunofluorescence, in growth-inhibition tests, and growth-precipitation tests. The type strains, together with the field strains, could be definitely identified by direct and indirect epi-immunofluorescence. In indirect epi-immunofluorescence, all rabbit antiserums against the strains of Mycoplasma hyorhinis tested by us always gave positive cross reactions with all the tested strains of Mycoplasma hyorhinis. On the other hand, in the growth-inhibition cross test and growth precipitation cross test, the type strains of Mycoplasma hyorhinis always reacted positively only with the homologous antiserums, whereas the heterologous reactions were mostly negative or dubious. The clearly positive epi-immunofluorescent reactions with serums against any strain of Mycoplasma hyorhinis suggest that this method is more suitable for the identification of Myocplasma species than the other two tests studied. 相似文献
15.
Six field strains of Mycoplasma hyopneumoniae isolated from pneumonic lungs of pigs, reference strains 11 and J of M hyopneumoniae, Ms 42 strain of Mycoplasma flocculare, and BTS 7 strain of Mycoplasma hyorhinis were compared serologically, using hyperimmune antisera produced in rabbits. All strains of M hyopneumoniae were closely related as determined with the disk growth-inhibition test; however, differences in zone sizes indicated that some antigenic heterogeneity existed. Cross-reactions were not detected between M hyopneumoniae, M flocculare, and M hyorhinis with the growth-inhibition test. The metabolic-inhibition test was more useful for detection of intraspecies antigenic difference than was the growth-inhibition test, since antigenic diversity was clearly detected among M hyopneumoniae strains. Slight cross-reactions were observed between M hyopneumoniae and M flocculare. Using 2-dimensional immunoelectrophoresis, antigenic differences were observed among M hyopneumoniae strains, although many common components also were detected in electropherograms. Mycoplasma flocculare possessed a close antigenic relationship to M hyopneumoniae, as determined by two-dimensional immunoelectrophoresis, whereas both organisms were less related to M hyorhinis. Evidence obtained in this study indicated that strains of mycoplasmas tentatively identified as M hyopneumoniae were similar antigenically, but evidence was obtained also of some diversity in antigenic structure among these strains. 相似文献
16.
Ogino S Munakata Y Ohashi S Fukui M Sakamoto H Sekiya Y Noormohammadi AH Morrow CJ 《Avian diseases》2011,55(2):187-194
Mycoplasma synoviae is an important causative agent of avian mycoplasmosis. In the present study the conserved domain of the variable lipoprotein and hemagglutinin (vlhA) gene of M. synoviae was sequenced and analyzed for 19 field strains of M. synoviae isolated from chickens across Japan. This analysis revealed that there were at least nine genotypes of M. synoviae present in Japan. Furthermore, we found a single nucleotide polymorphism (SNP) within this region in all the Japanese isolates, and based on this finding, we established a PCR method with cycling probe technology to differentiate between these field isolates and the live M. synoviae vaccine strain Mycoplasma synoviae-H (MS-H). 相似文献
17.
Rolf Jansson 《Acta veterinaria Scandinavica》1974,15(2):274
Eighteen strains of Mycoplasma hyorhinis and a strain of Mycoplasma suipneumoniae were tested in 4 serological tests, i. e., disc growth inhibition, metabolic inhibition, indirect haemagglutination and indirect epi-immunofluorescence. Only with immunofluorescence could all tested strains of M. hyorhinis be shown; no cross-reactions between M. hyorhinis and M. suipneumoniae could be detected. The other tests failed in many cases to identify strains of the same species, and they gave cross-reactions between M. hyorhinis and M. suipneumoniae. 相似文献
18.
Monitoring of susceptibility to antibiotics in field isolates of pathogenic avian mycoplasmas is important for appropriate choice of treatment. Our study compared in vitro susceptibility to enrofloxacin and difloxacin in recent (2005-2006) isolates of Mycoplasma gallisepticum and Mycoplasma synoviae from meat-type turkey flocks with archived (1997-2003) isolates and reference strains. Comparison of minimal inhibitory concentration (MIC) values determined by microtest, agar dilution and commercial Etest showed good agreement, but underscored the need for standardized methods for testing. Notably, while the commercial Etest was convenient and accurate for determining MICs for enrofloxacin in the range 0.002-0.094mug/ml, the endpoint of inhibition for M. gallisepticum and M. synoviae strains with MIC values >/=1.0mug/ml could not be determined. A decrease in susceptibility to both fluoroquinolones was detected in archived strains but to a greater degree in recent isolates, most of which had MICs above the NCCLS susceptibility breakpoint for these antibiotics (相似文献
19.
The aim of the study was to analyse experimental transmission of Mycoplasma synoviae, an avian pathogen. Three experiments using specific pathogen-free day-old chicks placed in isolators were conducted. In the first experiment, the birds were introduced in an isolator previously contaminated with a M. synoviae broth culture. After 34 days, these birds were eliminated and, for the second trial, the chicks were introduced in the same isolator without disinfecting. In the third assay, the chicks were placed in an isolator containing a mixture of food, feathers and dust collected less than an hour earlier from a M. synoviae infected laying hen flock. In the second and third experiments in order to exacerbate the M. synoviae infection, the birds were inoculated with infectious bronchitis (IB) virus. The presence of M. synoviae in the environment and in tracheal swabs was monitored by culture, a multiplex PCR (mPCR) detecting M. synoviae and Mycoplasma 16S rDNA and a multiplex RT-PCR (mRT-PCR) detecting the M. synoviae mRNA coding for a membrane protein and Mycoplasma 16S rRNA. In in vitro experimental conditions, M. synoviae mRNA and 16S rRNA were detected up to 20 min and 23 h respectively after mycoplasma death. In the first assay, the first infected bird was detected on the 13th day. In the second trial, culturable M. synoviae or viable M. synoviae were detected in the isolator for 3 or 4 to 5 days respectively after depopulation of the birds of the first assay whereas the first culture positive tracheal swabs were detected on the 33rd day, after IB inoculation. In the third experiment, the first infected birds were detected on the 54th day. Thus, the different assays showed that M. synoviae contaminated material (dust, feathers and food) can infect chicks, sometimes after remarkably long silent periods. 相似文献