Diversity among isolates of Actinobacillus equuli and related organisms as revealed by ribotyping

Objective The objective of this work was to examine the diversity within Australian isolates of Actinobacillus equuli and related organisms by the genotypic method of ribotyping.
Design Ribotyping, performed using the enzyme Hae III, was used to examine the diversity in 12 field isolates of A equuli (five being capable of fermenting L-arabinose), one field isolate of Pasteurella caballi and two unclassifiable field isolates. Isolates were obtained from Australian horses, except for three isolates of A equuli (one L-arabinose positive and two L-arabinose negative) which were obtained from horses and a pig in Africa. In addition, the type strains for A equuli and P caballi and a reference strain for Bisgaard Taxon 9 were included in the study.
Results The ribotype patterns were analysed by computerised cluster analysis, yielding five clusters (A to E). All five of the L-arabinose positive A equuli were assigned to cluster A, with all the other seven A equuli isolates (all L-arabinose negative) and the type strain being assigned to cluster B. One of the two unclassified isolates formed cluster C along with the reference strain for Bisgaard Taxon 9. The remaining unclassified isolate formed cluster D. Cluster E consisted of the field isolate and reference strain of P caballi .
Conclusion The results of this study indicate that A equuli is a diverse species, with L-arabinose positive isolates of A equuli being quite distinct from typical L-arabinose negative isolates. Ribotyping appears to be a useful tool in confirming the identity of A equuli -like organisms from horses.     

柱花草种间亲缘关系SSR分析

为研究柱花草(Stylosanthes spp.)种间的亲缘关系,本研究利用23个SSR标记对柱花草属14个种的33份材料的遗传关系进行了研究。结果表明,23个SSR标记在不同柱花草种间均具有多态性,共检测到等位基因138个,每个标记可检测到4～10个等位基因,平均6个;每个标记的多态性信息含量在0.501～0.830,平均为0.700。聚类分析和主成分分析结果表明, 33份供试柱花草种质可分为5类,根据包含的基因组类型分别命名为AB类、AC类、J类、DE类、G类。毛叶柱花草(S. subsericea)和有钩柱花草(S. hamata)(I)的亲缘关系最近,有钩柱花草(I)与头状柱花草(S. capitata)的遗传关系较远。     

结缕草属植物种间关系和遗传多样性的SRAP标记分析

 通过ＳＲＡＰ标记技术对结缕草属植物５种１变种共９６份种源进行遗传多样性分析。结果表明,１）结缕草属种质间存在丰富的遗传多样性,５４对ＳＲＡＰ引物共扩增出３６２条清晰的用于多样性分析的谱带,其中多态性条带３３７条,多态性比率（ＰＰＢ）为９３．０９％,Ｎｅｉ’ｓ基因多样性指数（犎）为０．２４０９,Ｓｈａｎｎｏｎ信息指数（犐）为０．３７４６。２）应用Ｎｅｉ－Ｌｉ相似系数法估算了９６份材料间的遗传相似系数（ＧＳ）,其ＧＳ为０．５９３９～０．９８３４,平均为０．７５８８。不同种之间,结缕草与中华结缕草、沟叶结缕草与细叶结缕草中的部分材料遗传相似系数较高,遗传距离较近。大穗结缕草Ｚ０１０和长花结缕草Ｚ１２２与其他结缕草的遗传相似系数都相对较低。同一个种内,结缕草、中华结缕草和沟叶结缕草种内遗传变异均较大,ＧＳ分别为０．５９９４～０．９８３４,０．６２４３～０．９８０７和０．６６３０～０．９４７５。３）通过ＵＰＧＭＡ 分子系统聚类法,将９６份种源分为７个大的类群,其中第１大类和第２大类均包含结缕草、中华结缕草和少量的沟叶结缕草种源;第３大类群主要包括４份美国引进的材料;第４大类主要包括沟叶结缕草和细叶结缕草;大穗结缕草Ｚ０１０、长花结缕草Ｚ１２２和结缕草Ｚ１１５都单独聚为一类。从聚类结果可以看出,结缕草、中华结缕草和沟叶结缕草均有交叉聚类现象,并不是同一个种的材料完全相聚,个别种源自行一类,遗传基础与其他材料差异较大,从遗传聚类图可以很明确地看出９６份种源间的遗传距离及亲缘关系。     

The C-terminal variable domain of LigB from Leptospira mediates binding to fibronectin

Adhesion through microbial surface components that recognize adhesive matrix molecules is an essential step in infection for most pathogenic bacteria. In this study, we report that LigB interacts with fibronectin (Fn) through its variable region. A possible role for LigB in bacterial attachment to host cells during the course of infection is supported by the following observations: (i) binding of the variable region of LigB to Madin-Darby canine kidney (MDCK) cells in a dose-dependent manner reduces the adhesion of Leptospira, (ii) inhibition of leptospiral attachment to Fn by the variable region of LigB, and (iii) decrease in binding of the variable region of LigB to the MDCK cells in the presence of Fn. Furthermore, we found a significant reduction in binding of the variable region of LigB to Fn using small interfering RNA (siRNA). Finally, the isothermal titration calorimetric results confirmed the interaction between the variable region of LigB and Fn. This is the first report to demonstrate that LigB binds to MDCK cells. In addition, the reduction of Fn expression in the MDCK cells, by siRNA, reduced the binding of LigB. Taken together, the data from the present study showed that LigB is a Fn-binding protein of pathogenic Leptospira spp. and may play a pivotal role in Leptospira-host interaction during the initial stage of infection.     

四川常羽乌骨鸡群体遗传多样性分析

利用选自家禽基因组的10个微卫星标记,对四川常羽乌骨鸡5个群体(四川山地乌骨鸡白羽系、黑羽系;黄忠山地乌骨鸡;黄忠山地乌骨鸡绿壳蛋系;草科乌骨鸡)的遗传多样性进行了检测,计算了各群体的群体杂合度、群体间遗传距离,并根据遗传距离进行了聚类分析。结果表明所选择的微卫星标记在各群体中表现出较高的多态性;四川5个乌骨鸡群体遗传多样性比较丰富,群体平均杂合度为0.5383到0.6659;各群体间的遗传距离有一定的差异;聚类分析将5个群体聚为三类。     

Genetic components in contemporary Faroe Islands Cattle as revealed by microsatellite analysis

The gene pool of indigenous Faroe Islands Cattle is strongly affected by crossing with Norwegian Red. In this study, the genetic structure in 191 animals representing five North European cattle breeds (Faroe Islands, Icelandic, Blacksided Troender, Western Fjord and Norwegian Red) and the genetic admixture in the contemporary Faroe Islands Cattle at the population and individual level were evaluated using 20 polymorphic microsatellite loci. Only 6.7% of the total genetic variation could be attributed to the differences amongst the breeds. The factorial correspondence analysis based on all allele frequencies could hardly reveal a divergence between Faroe Islands Cattle and Norwegian Red. In addition, a Neighbor‐Net tree constructed to examine the allocation of individuals of Faroe Islands Cattle and Norwegian Red provided a detailed interrelationship network for all the 72 animals. An estimation of the population admixture proportion showed a strong genetic contribution by Norwegian Red (47.3%) in the contemporary Faroe Islands Cattle. On the other hand, individual admixture analysis demonstrated that only seven of the individual Faroe Islands Cattle analysed, which also showed more traditional colour patterns, could be assigned to the Faroe Islands Cattle cluster, probably representing the breed's remaining purebred animals. Strategies for preserving the original native genes in Faroe Islands Cattle should be considered in order to prevent the breed from becoming extinct and to strengthen the breed's capability in future breeding programmes.     

Measurement of canine plasma fibronectin by rocket immunoelectrophoresis

Plasma fibronectin concentrations of 148 normal canine samples were measured by rocket immunoelectrophoresis. Electrophoresis was accomplished, using 2.0% rabbit anticanine fibronectin by volume in 0.7% agarose in buffer. Films were electrophoresed 18 hours in barbital buffer, 7.5 mA/film. The mean fibronectin concentration for normal citrated dog plasma was 290 micrograms/ml +/- 50 micrograms/ml.     

基于叶绿体atpB-rbcL序列探讨猬草属和赖草属植物的系统发育和母系起源

对小麦族猬草属、赖草属(NsXm)、披碱草属(StH)及含不同染色体组(St、H、Ns、E^b、E^e、P、F 、V、W)的近缘二倍体属共46个类群的叶绿体基因间隔区atpB-rbcL序列进行系统发育分析,探讨猬草属和赖草属植物系统发育关系及母系起源。结果显示,1)猬草属模式种H. patula与拟鹅观草属和披碱草属物种聚为一支,表明H. patula与披碱草属植物亲缘关系近,其母本来源为含St染色体组的拟鹅观草属物种;2)猬草、长芒猬草、4个新麦草属物种及滨麦和所有欧亚分布的赖草属物种聚在一支,说明猬草、长芒猬草与欧亚赖草亲缘关系较近,其母本供体来自含Ns染色体组的新麦草属物种;3)高丽猬草和东北猬草与北美赖草及冰草属、旱麦草属、大麦属物种处于同一分支,表明高丽猬草和东北猬草与北美赖草具有较近亲缘关系,其母本供体为未知来源的Xm染色体组。研究结果支持将H. patula组合到披碱草属中,猬草、长芒猬草、高丽猬草和东北猬草应组合到赖草属中。     

Heterogeneity of Mycoplasma iowae determined by restriction enzyme analysis

Strains of Mycoplasma iowae were homogeneous in some characteristics and heterogeneous in others. Thus, the biochemical tests, immunofluorescence, and protein profiling by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were group-specific tests. However, some minor differences in protein patterns were seen among strains. The growth inhibition test tended to be strain-specific. Hemagglutination titers were very low and unstable for the majority of strains. One strain (RY-65) with a stable high-titer hemagglutinin failed to react in the hemagglutination-inhibition test against immune sera to the reference strains. Restriction endonuclease DNA analyses was the most useful method to differentiate 1 strain from another.     

The morphology of the normal end-plate in lambs as revealed by silver impregnation and light microscopy.

A technique employing silver impregnation can be used on frozen formalin-fixed material for demonstrating ovine motor end-plates. The morphology of ovine motor end-plates is variable but broadly similar to that described in other species. The end-plates are oval with approximate dimensions of 33 X 17 micrometer. The distal ramifications of the nerve fibre show terminal and sub-terminal vesicles. The endings are flimsy or robust and the latter have single, dual or multiple terminations.     

低氮胁迫对高羊茅IAA生长素代谢组影响的研究

氮是植物生长发育必须的大量元素之一,增施氮肥虽然有利于植物产量的提高,但是不利于土壤的可持续利用。牧草类作物相对于其他农作物受贫瘠土壤胁迫的危害更加严重,然而相应的理论研究还比较滞后。本文以禾本科牧草资源高羊茅为研究对象,利用LC-ESI-MS(液相色谱电离串联质谱)分析了正常和低氮条件下高羊茅叶片中的代谢组分,两种材料中分别稳定检测到1424和1251种代谢产物,利用OPLS-DA(正交偏最小二乘法判别分析)分析方法,鉴别到低氮处理后主要有13种代谢物质的含量发生变化,其中包括了生长素IAA。利用荧光定量PCR的方法检测8个IAA信号途径相关基因的表达,发现其表达在低氮处理后均被诱导,另外通过测定IAA激素的含量,也证实了IAA在高羊茅缺氮处理后升高这一事实。本研究为高羊茅氮缺乏的分子机理提供了重要的线索,也为在生产中通过喷施IAA来缓解植物应对氮缺乏的措施提供了理论依据。     

High genetic relatedness among Mycobacterium avium strains isolated from pigs and humans revealed by comparative IS1245 RFLP analysis

Members of the Mycobacterium avium complex cause pig mycobacteriosis and opportunistic human infections. Infections due to environmental mycobacteria are increasing in both industrial and developing countries. Mycobacterium-infected pig carcasses can pass for human consumption due to the poor specificity of meat control by visual detection at the slaughter houses. The genetic relatedness of porcine and human MAC isolates in Finland has been unknown. M. avium isolates isolated from pig organs (n=16) and clinical samples (n=13) were compared by IS1245 RFLP analysis to evaluate the similarity of the isolates obtained from human and porcine samples. Nearly identical multicopy M. avium subsp. hominissuis IS1245 RFLP fingerprints were obtained for isolates of porcine and human origin. IS1245 RFLP patterns of 38% of the porcine and human M. a. hominissuis isolates were >90% similar. The RFLP patterns of two porcine and two human isolates showed >95% similarity. The high similarity of the IS1245 RFLP patterns of the human and porcine M. a. hominissuis isolates indicates close genetic relatedness, suggesting that M. a. hominissuis is transmitted between pigs and humans, or that pigs and humans share common environmental sources of infection. Porcine and human isolates with RFLP patterns differing by only one or two bands were found, which shows that the same M. a. hominissuis strains may infect both humans and pigs.