Characterization of Borrelia Species Isolated from Ixodid Ticks,Ixodes ovatus

Thirty-two Borrelia isolates were obtained from the adult stage of ixodid ticks, Ixodes ovatus, collected in various localities in Japan. Borrelial isolates were cultivated and analyzed by polyacrylamide gel electrophoresis, with monoclonal antibodies, by pulsed field gel electrophoresis, and by genomic Southern hybridization. All borrelial isolates showed similar protein profiles and monoclonal antibody reactivities, while plasmid profiles were rather diverse. Genomic hybridization using rRNA gene probes demonstrated the genetic similarities of those isolates. We found no significant differences among the borrelial isolates tested, and the restriction fragment length polymorphism patterns of I. ovatus isolates were quite distinct from those of borrelial strains associated with Lyme disease. Therefore, the isolates of Borrelia obtained from I. ovatus were thought to fall into different genospecies.     

Genomic Analysis of Borrelia japonica Sp. Nov. Isolated from Ixodes ovatus in Japan

Genetic characteristics of 12 Borrelia isolates from the tick, Ixodes ovatus, I. persulcatus, and the rodent, Apodemus speciosus ainu, in Japan were compared to members of the three genospecies of Borrelia burgdorferi sensu lato; B. burgdorferi sensu stricto, B. garinii and group VS461. The methods used in this study were the quantitative microplate DNA hybridization assay and restriction fragment length polymorphism (RFLP) analyses of the flagellin structural genes and the 16S rRNA genes. The six isolates from I. persulcatus and A. speciosus ainu were identified as genospecies B. garinii using RFLP analysis of the flagellin and 16S rRNA genes. In contrast, RFLP analysis of the six isolates from I. ovatus indicated that they were different from the three reported genospecies. DNA homology studies confirmed the RFLP results. The six isolates from I. ovatus had DNA homologies ranging from 85 to 99%, whereas DNA relatedness of the I. ovatus isolate with strains belonging to the three genospecies was 50 to 64%. These results suggest that the strains isolated from I. ovatus in Japan differ from the three genospecies and should be classified as a new genospecies of B. burgdorferi sensu lato. We propose that strains isolated from I. ovatus should be classified as B. japonica sp. nov.     

Patterns and origin of intraspecific functional variability in a tropical alpine species along an altitudinal gradient

Background : Intraspecific functional variability (IFV) along altitudinal gradients is a powerful proxy to infer the responses of plants to abrupt environmental changes. We envisage that IFV shows distinctive patterns in tropical and extratropical alpine regions.

Aims : To characterise the patterns and explore the origin of IFV in a tropical alpine species in a context of upward range extension.

Methods : We examined variations in a series of plant functional traits in Lasiocephalus ovatus, inside and outside a nurse plant along a 600 m altitudinal gradient in the Ecuadorian Andes, and we studied its genetic variability.

Results : More conservative traits were developed at higher elevation, in contrast to extratropical alpine plants, which commonly develop opportunistic traits in response to late snowmelt close to their upper altitudinal limit. The presence of nurse cushions did not alter this trend. Increasing genetic distance along the gradient suggested that IFV might be partly genetically induced.

Conclusions : Our data combined with existing literature in tropical alpine environments lead the way to a stimulating scientific challenge: determining if patterns of plant altitudinal distribution in tropical alpine areas in response to climate change are predictable from patterns described in extratropical alpine areas.     

Establishment and characterization of a cell line from the head kidney of golden pompano Trachinotus ovatus and its application in toxicology and virus susceptibility

A cell line derived from the head kidney of golden pompano Trachinotus ovatus (TOHK) was established and characterized in this study. The TOHK cells grew most rapidly at 28° C and the optimum foetal bovine serum concentration in L‐15 medium was 10%. The TOHK cells have a diploid chromosome number of 2N = 54. The transfection efficiency of TOHK cells was 7·5% at the 15th passage and 72% at the 40th passage. The transfection efficiency in TOHK cells was high, so these cells are suitable for foreign gene expression. The cytotoxic effects of heavy metals and extracellular products from Vibrio anguillarum and Vibrio alginolyticus were demonstrated in TOHK cells, so this TOHK cell line could also be applied in environmental monitoring of heavy metals and pathogenic bacteria. TOHK cell line showed high virus susceptibility, such as grouper nervous necrosis virus (GNNV) and Singapore grouper iridovirus (SGIV). Then, TOHK cell line could be used for the study of viral pathogenesis and the development of antiviral strategies.     

腥黑粉菌属一新种

本文报道了从意大利进口兔尾草(Lagurus ovatus L.)上截获的腥黑粉菌属一新种——兔尾草腥黑粉菌(Tilletia laguri G.M.Zhang,G.X.Lin & J.R.Deng),同时讨论了该种与近似种的区别,文中对其形态特征有拉丁文和汉文描述,并附有图片。主模式标本保存在深圳动植物检疫局真菌标本室,等模式标本分别保存在德国蒂宾根大学范基黑粉菌标本馆(HUV)、中国科学院微生物研究所真菌标本室和华南农业大学植保系真菌标本室。     

椭圆食粉螨线粒体DNA CO Ⅰ基因片段序列分析

采用苯酚-氯仿-异戊醇抽提法提取了采自江西南昌和广州潮州2个地理种群的椭圆食粉螨Aleuro-glyphus ovatus Troupeau,1878基因组DNA。以相应引物对椭圆食粉螨线粒体DNA细胞色素氧化酶I亚基基因(mtDNA COI)进行PCR扩增,直接测序,得到379bp的碱基片断(国际基因库索引号EF527825),其碱基序列A、C、G、T含量分别为71bp(18.73%)、53bp(13.98%)、89bp(23.48%)、166bp(43.80%);江西南昌和广州潮州2个地理种群间的椭圆食粉螨mtDNACOI基因片段完全一致,未发现地理差异。     

普通肉食螨对椭圆食粉螨的捕食功能

在6个恒温下研究普通肉食螨Cheyletus eruditus(Schrank)不同螨态对椭圆食粉螨Aleuroglyphus ovatus(Troupeau)的功能反应。结果表明,普通肉食螨不同螨态对椭圆食粉螨的功能反应均属于HollingⅡ型,其中雌成螨的捕食能力最强,其次是雄螨、若螨、幼螨;在各温度处理中,雌成螨在28℃时具有较高的捕食功能;普通肉食螨在16℃的低温状态下捕食功能很低,仅有雌螨对猎物有捕食行为;在猎物密度不变的情况下,普通肉食螨捕食猎物的数量随自身密度的增加而下降。     

Japanese Babesia microti cytologically detected in salivary glands of naturally infected tick Ixodes ovatus

Babesia microti protozoa were detected by light and electron microscopy in the salivary glands of field-collected Ixodes ovatus ticks; 6 of 85 adult ticks were demonstrated to be positive for B. microti DNA by polymerase chain reaction assays. In the salivary glands of unfed ticks, B. microti existed in the sporoblast stage in the granular acinus cells, and developed into the sporozoite stage during feeding on the host for 2 days. The present results indicated for the first time that I. ovatus can indeed carry B. microti and is not infected mechanically with the parasites by blood-sucking. This frequent infection of I. ovatus with B. microti demonstrates the significance of such a vector-pathogen relationship in Japan, and strongly suggests that I. ovatus is involved in the maintenance of B. microti in the fauna of Japanese rodents.     

卵形鲳鯵对刺激隐核虫的免疫应答和免疫保护研究

用刺激隐核虫(Cryptocaryon irritans)的幼虫对卵形鲳鯵(Trachinotus ovatus)进行腹腔注射和体表感染,然后每隔一周用阻动试验(Immobilization assay)检测免疫鱼的抗血清和皮肤培养液对刺激隐核虫幼虫的阻动效价,在第14周中,分别用亚致死剂量和致死剂量的刺激隐核虫幼虫对免疫鱼攻毒以检测所产生的免疫保护力。实验结果显示:两种免疫方法都能让卵形鲳鯵的血清和皮肤生成阻动刺激隐核虫幼虫的特异性抗体,并能使被免疫鱼获得明显的免疫保护,但是体表感染免疫组的血清和皮肤培养液的阻动效价都要比腹腔注射免疫组高,所获得的免疫保护力也更强。同时还发现,免疫鱼血清和皮肤培养液中的抗体存在明显的差异:两者的最初生成时间、达到峰值的时间、变化规律以及阻动效价等都不一致。因此,我们推测鱼类的系统免疫应答和皮肤黏膜免疫应答有可能是相互独立的,或者是不同步的。鱼类的体液免疫应答,特别是黏膜免疫应答对抵御刺激隐核虫的感染起了重要的作用,采用刺激隐核虫虫体疫苗可能成为预防海水鱼类白点病的一种选择。       

卵形鲳鲹胚胎及早期仔鱼耗氧量的研究

运用SKW-3微量呼吸仪对卵形鲳鲹(Trachinotus ovatus)胚胎和早期仔鱼的耗氧量进行测定,研究温度、盐度及pH的变化对胚胎耗氧量和Cu²⁺、Cd²⁺等重金属离子对早期仔鱼的耗氧量的影响。结果表明:在(25±0.5)℃条件下,卵形鲳鲹胚胎和早期仔鱼的耗氧量均随着发育时间的延长总体上呈上升的趋势,胚胎耗氧量在出膜前达到最大值,其中以原肠期和出膜期耗氧量变化最为显著。胚胎耗氧量随着温度、盐度、pH的增大而逐渐升高,在水温25℃、盐度35和pH为8时耗氧量到达最大值,随后逐渐减小。随着Cu²⁺浓度的升高,早期仔鱼耗氧量呈先增大后减小,在0.01mg·L^-1时耗氧率最大:胚胎耗氧量随着Cd²⁺浓度的升高而逐渐降低。