Recombination, Diversity and Allele Sharing of Infectivity Proteins Between Bartonella Species from Rodents

The alpha-Proteobacterium Bartonella is a common parasite of voles and mice, giving rise to short-lived (4 weeks to 2 months) infections. Here, we report high sequence diversity in genes of the VirB/VirD type IV secretion system (T4SS), amongst Bartonella from natural rodent populations in NE Poland. The VirB5 protein is predicted to consist of three conserved alpha helices separated by loops of variable length which include numerous indels. The C-terminal domain includes repeat stretches of KEK residues, reflecting underlying homopolymeric stretches of adenine residues. A total of 16 variants of VirB5, associated with host identity, but not bacterial taxon, were identified from 22 Bartonella isolates. One was clearly a recombinant from two others, another included an insertion of two KEK repeats. The virB5 gene appears to evolve via both mutation and recombination, as well as slippage mediated insertion/deletion events. The recombinational units are thought to be relatively short, as there was no evidence of linkage disequilibrium between virB5 and the bepA locus only 5.5 kb distant. The diversity of virB5 is assumed to be related to immunological role of this protein in Bartonella infections; diversity of virB5 may assist persistence of Bartonella in the rodent population, despite the relatively short (3-4 weeks) duration of individual infections. It is clear from the distribution of virB5 and bepA alleles that recombination within and between clades is widespread, and frequently crosses the boundaries of conventionally recognised Bartonella species.     

The Distribution and Diversity of Bartonella Species in Rodents and Their Ectoparasites across Thailand

Our study highlights the surveillance of Bartonella species among rodents and their associated ectoparasites (ticks, fleas, lice, and mites) in several regions across Thailand. A total of 619 rodents and 554 pooled ectoparasites (287 mite pools, 62 flea pools, 35 louse pools, and 170 tick pools) were collected from 8 provinces within 4 regions of Thailand. Bandicota indica (279), Rattus rattus (163), and R. exulans (96) were the most prevalent species of rats collected in this study. Real-time PCR assay targeting Bartonella-specific ssrA gene was used for screening and each positive sample was confirmed by PCR using nuoG gene. The prevalence of Bartonella DNA in rodent (around 17%) was recorded in all regions. The highest prevalence of Bartonella species was found in B. savilei and R. rattus with the rate of 35.7% (5/14) and 32.5% (53/163), respectively. High prevalence of Bartonella-positive rodent was also found in B. indica (15.1%, 42/279), and R. norvegicus (12.5%, 5/40). In contrast, the prevalence of Bartonella species in ectoparasites collected from the rats varied significantly according to types of ectoparasites. A high prevalence of Bartonella DNA was found in louse pools (Polyplax spp. and Hoplopleura spp., 57.1%) and flea pools (Xenopsylla cheopis, 25.8%), while a low prevalence was found in pools of mites (Leptotrombidium spp. and Ascoschoengastia spp., 1.7%) and ticks (Haemaphysalis spp., 3.5%). Prevalence of Bartonella DNA in ectoparasites collected from Bartonella-positive rodents (19.4%) was significantly higher comparing to ectoparasites from Bartonella-negative rodents (8.7%). The phylogenetic analysis of 41 gltA sequences of 16 Bartonella isolates from rodent blood and 25 Bartonella-positive ectoparasites revealed a wide range of diversity among Bartonella species with a majority of sequences (61.0%) belonging to Bartonella elizabethae complex (11 rodents, 1 mite pool, and 5 louse pools), while the remaining sequences were identical to B. phoceensis (17.1%, 1 mite pool, 5 louse pools, and 1 tick pool), B. coopersplainensis (19.5%, 5 rodents, 1 louse pool, and 2 tick pools), and one previously unidentified Bartonella species (2.4%, 1 louse pool).     

Prevalence and Genetic Diversity of Bartonella spp. in Small Mammals from Southeastern Asia

Among 1,341 blood samples from rodents that were trapped in Southeast Asia between 2008 and 2010, we found a prevalence of Bartonella infection ranging from 9.6 to 11.9%. Bartonella species identified (143 isolates) included B. elizabethae, B. coopersplainsensis, B. phoceensis, B. queenslandensis, B. rattimassiliensis, B. tribocorum, and three new putative Bartonella species.     

Prevalence and Diversity of Small Mammal-Associated Bartonella Species in Rural and Urban Kenya

Several rodent-associated Bartonella species are human pathogens but little is known about their epidemiology. We trapped rodents and shrews around human habitations at two sites in Kenya (rural Asembo and urban Kibera) to determine the prevalence of Bartonella infection. Bartonella were detected by culture in five of seven host species. In Kibera, 60% of Rattus rattus were positive, as compared to 13% in Asembo. Bartonella were also detected in C. olivieri (7%), Lemniscomys striatus (50%), Mastomys natalensis (43%) and R. norvegicus (50%). Partial sequencing of the citrate synthase (gltA) gene of isolates showed that Kibera strains were similar to reference isolates from Rattus trapped in Asia, America, and Europe, but that most strains from Asembo were less similar. Host species and trapping location were associated with differences in infection status but there was no evidence of associations between host age or sex and infection status. Acute febrile illness occurs at high incidence in both Asembo and Kibera but the etiology of many of these illnesses is unknown. Bartonella similar to known human pathogens were detected in small mammals at both sites and investigation of the ecological determinants of host infection status and of the public health significance of Bartonella infections at these locations is warranted.     

醋栗番茄Solanum pimpinellifolium遗传多样性分析

野生种醋栗番茄包含丰富的变异,具有众多优良性状。本研究对从世界不同地区收集的433份醋栗番茄遗传资源进行了表型鉴定及遗传多样性分析。表型鉴定结果表明,在收集的433份醋栗番茄中约有14%为樱桃番茄,370份典型醋栗番茄中约22%的材料存在不同程度的分离。群体变异分析表明,不同性状间变异系数存在较大差异,其中柱头变异系数最大,为56.716%;花瓣数遗传稳定,变异系数最小,为2.082%;相关分析表明,多个性状间存在显著相关性;利用表型和基因型数据聚类均将醋栗番茄群体划分为两大类群;主成分分析表明,坐果率、单果重、可溶性固形物对变异的贡献率较大。研究结果将为利用醋栗番茄进行栽培种番茄遗传改良奠定一定的基础。     

Prevalence of Antibodies to Borrelia Spirochetes among Wild Small Rodents in Central and Western Japan

Field rodents serve as a reservoir for Lyme disease spirochetes. To evaluate the antibody responses of rodents against different Borrelia species in relation to fauna of vector ticks feeding on them, we examined 272 sera of wild rodents, Apodemus speciosus, A. argenteus, and Eothenomys smithii, obtained in 27 locations in central and western Japan from 1981 to 1994. As to prevalences by rodent species using immunoperoxidase test, A. speciosus, A. argenteus and E. smithii showed 29.4%, 11.6% and 30.8% reactivity to Borrelia japonica, 10.7%, 7.2% and 3.8% to B. afzelii, 0.6%, 1.4% and 0% to B. garinii, and 14.7%, 7.2% and 11.5% to an unknown Borrelia species designated as It type, respectively. Each antibody to B. japonica, B. afzelii and B. sp. It type was detected widely both in central and western Japan, but the antibody to B. garinii was scarcely detectable in any area and rodent species examined. Apodemus mice in high mountain altitudes tend to have antibody to B. afzelii or B. japonica, and those in lower altitudes tend to have B. japonica or B. sp. It type. All 13 Apodemus mice from which B. japonica or B. sp. It type were isolated showed higher titers of antibodies to each homologous Borrelia species. The present results indicate that these antibody prevalences among rodents may be associated with dominant Ixodes ovatus and sporadic I. persulcatus on the mainland of Japan, and that Apodemus mice may not be an efficient reservoir for B. garinii. Such a serosurvey is a useful measure to evaluate the natural distribution of the pathogen.     

Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia

Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions—exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.     

甘蔗近缘种滇蔗茅(Erianthus rockii)表型性状遗传多样性研究

为有效评价和利用蔗茅种质资源,挖掘其优良性状,以滇、黔、川考察收集的29份蔗茅为材料,选取5个数量性状为指标,对其多样性指数、变异系数、数量性状之间的相关性、数量性状与经纬度、海拔的相关性进行研究。分析结果表明：(1)共采集到25份高海拔种质资源,其中海拔超过2800米有4份,进一步丰富了甘蔗野生种质资源库;(2)蔗茅种质资源数量性状的Shannon-Wiener多样性指数均较高,株高多样性指数最高(1.441),锤度多样性指数最低(1.291);数量性状遗传变异较丰富,各性状变异系数范围为21%～38%,变异系数最大的为株高(38%),最小的为叶长(21%);(3)蔗茅种质资源性状差异显著,叶长、叶宽、株高、茎径存在较大相关性,而与锤度不相关;叶宽与经度呈正相关,与纬度、海拔呈负相关。(4)聚类分析结果表明,蔗茅种质资源可分为四大类群,其中Ⅱ类群(EF-27)具有良好的数量性状和锤度品质潜力,可推荐作为甘蔗育种杂交利用首选材料。     

云南48种木兰科野生植物资源遗传多样性研究

为了解云南省木兰科（Magnoliaceae）野生植物资源的遗传多样性,利用ISSR分子标记技术对48种木兰科野生植物资源进行研究。结果表明,10对引物共扩增出151条带,均为多态性条带,多态性条带百分率为100%。总的观测等位基因数（N_a）为2.000 0,有效等位基因数（N_e）为1.564 5,Nei’s基因多样性指数（H）0.337 9,Shannon’s信息指数（I）为0.510 1。总的基因多样性指数（H_t）为0.368 0,属间基因多样性指数（D_st）为0.251 9,占68.4%,基因分化系数（G_st）为0.684 0,基因流（N_m）为0.231 0。UPGMA聚类分析将48种木兰科植物划分为7个类群,各类群并非按照属聚在一起,而是不同属植物相间分布,长喙厚朴（Magnolia rostrata）、素黄含笑（Michelia flaviflora）和球花含笑（M.sphaerantha）可能为云南省木兰科植物中的原始种。48种木兰科野生植物总体具有较高的遗传多样性,但属间遗传变异较高,基因流较小,存在遗传漂变的风险,聚类结果与刘玉壶的分类系统存在分歧,这从分子水平为木兰科植物间的起源、进化与分类提供了重要依据。     

甘蔗近缘种滇蔗茅(Erianthus rockii)表型性状遗传多样性研究

选用6个数量性状和23个质量性状对滇蔗茅野生资源的表型变异、多样性及聚类关系进行分析。结果表明,数量性状变异系数在12.72%~22.38%之间,最大的是田间锤度,为22.38%,其次是茎径,为22.27%,最小的是叶长,为12.72%;质量性状的曝光后节间颜色多样性丰富,茎形、芽形、生长裂缝、节间形状、花序形状等5个性状均表现一致;聚类分析表明51份滇蔗茅无性系材料可分为4大类群和5个亚类群。遗传多样性和聚类分析为资源杂交利用和优异基因挖掘提供参考。     

High Prevalence and Species Diversity of Helicobacter spp. Detected in Wild House Mice

PCR diagnostics detected 100% prevalence of Helicobacter in 425 wild house mice (Mus musculus) from across central Europe. Of seven species identified, the five most frequent were Helicobacter rodentium (78%), H. typhlonius (53%), H. hepaticus (41%), H. bilis (30%), and H. muridarum (1%). Double infections were more common (42%) than single (30%) and triple (21%) infections. Wild house mice could be considered potential reservoirs of Helicobacter strains for both humans and other vertebrates.     

云南杜鹃花上的酵母菌物种多样性

利用酵母菌常规分类学方法 ,对采自云南省东部、西部及中部地区杜鹃花上的 119株酵母菌进行了分类研究 ,以期弄清其属种分布特点 ,从而为发掘新种、开发酵母菌资源提供依据。这些酵母菌被归入3大类 10个属 ,显示了云南杜鹃花中酵母菌的种类多样性 ,其中优势属为假丝酵母属 (Candida)。     

Recombination Within and Between Species of the Alpha Proteobacterium Bartonella Infecting Rodents

Bartonella infections from wild mice and voles (Apodemus flavicollis, Mi. oeconomus, Microtus arvalis and Myodes glareolus) were sampled from a forest and old-field habitats of eastern Poland; a complex network of Bartonella isolates, referrable to B. taylorii, B. grahamii, B. birtlesii and B. doshiae, was identified by the sequencing of a gltA fragment, comparable to previous studies of Bartonella diversity in rodents. Nested clade analysis showed that isolates could be assigned to zero- and one-step clades which correlated with host identity and were probably the result of clonal expansion; however, sequencing of other housekeeping genes (rpoB, ribC, ftsZ, groEl) and the 16S RNA gene revealed a more complex situation with clear evidence of numerous recombinant events in which one or both Bartonella parents could be identified. Recombination within gltA was found to have generated two distinct variant clades, one a hybrid between B. taylorii and B. doshiae, the other between B. taylorii and B. grahamii. These recombinant events characterised the differences between the two-step and higher clades within the total nested cladogram, involved all four species of Bartonella identified in this work and appear to have played a dominant role in the evolution of Bartonella diversity. It is clear, therefore, that housekeeping gene phylogenies are not robust indicators of Bartonella diversity, especially when only a single gene (gltA or 16S RNA) is used. Bartonella clades infecting Microtus were most frequently involved in recombination and were most frequently tip clades within the cladogram. The role of Microtus in influencing the frequency of Bartonella recombination remains unknown.     

Genome Sequence of Bartonella birtlesii, a Bacterium Isolated from Small Rodents of the Genus Apodemus

Bartonella birtlesii is a facultative intracellular bacterium isolated from the blood of small mammals of the genus Apodemus. The present study reports the draft genome of Bartonella birtlesii strain IBS 135(T) (CIP 106691(T)).     

湖南野生食用菌物种多样性研究

本次报道湖南野生食用菌资源265种,这是近40年来的考察研究结果.这些种类中包括子囊菌10种,担子菌255种,许多种类具有重要的利用和开发意义.对每个种的生态特征和利用价值均进行了简单介绍.     

广东5个野生大豆居群的遗传多样性分析

采用SSR分子标记对来自广东省5个县野生大豆居群的遗传多样性进行了分析,为广东野生大豆资源保护和利用提供依据。结果显示:(1)5个野生大豆居群在60个SSR位点共检测出263个等位变异,同一位点上等位基因数目最多为10个,最少为2个,平均为4.38个;不同群体中特有等位基因数不同,其中连州(LZ)和南雄(NX)野生大豆的特有等位基因数目较多,分别为19个和16个;Shannon指数(I)变化范围为0.162~2.174,期望杂合度(He)的变化范围为0.073~0.899。(2)广东连州(LZ)野生大豆居群的遗传多样性最高,而仁化(RH)野生大豆的遗传多样性最低,二者的Shannon指数(I)分别为0.811、0.113;群体分子方差(AMOVA)分析结果揭示,居群间变异占51%,群体内变异占49%,且仁化居群与其他居群间基因流较小。(3)依据遗传距离可将连州(LZ)和连南(LN)聚类为一类,乳源(RY)和南雄(NX)为一类,仁化(RH)单独为一类。研究表明,广东不同野生大豆居群间遗传多样性差异较大,而且居群内基因类型多,其中连州(LZ)和乳源(RY)野生大豆居群的遗传多样性较高,证明广东野生大豆群体保存了丰富的基因资源。     

Blastocystis sp. Subtype Diversity in Wild Carnivore Species from Spain

The occurrence and molecular diversity of the stramenopile eukaryote Blastocystis sp. was investigated by PCR and sequencing (Sanger and NGS) methods in 380 faecal specimens of free‐living carnivores in Spain. Blastocystis sp. was confirmed in 1.6% (6/380) of the specimens analysed. Two samples from a common genet and a fox were successfully subtyped as ST7 by Sanger. Using NGS, ST14 was found in a fox and a European polecat, ST7 in a fox, and two additional foxes presented mixed infections of ST1/ST2/ST4 and ST1/ST2/ST7, respectively. Wild carnivore species could act as carriers of zoonotic Blastocystis subtypes.     

Diversity of Fusarium Species Isolated from Weeds and Plant Debris in Croatia

Weeds are alternative hosts of plant pathogens and when colonized may not exhibit disease symptoms. In 2008 and 2009, samples of weeds and plant debris were collected from 12 locations in eastern Croatia, and 300 Fusarium isolates colonizing them were identified. Strains were grouped and identified based on morphology and amplified fragment length polymorphism (AFLP) patterns. Portions of the β‐tubulin and translocation elongation factor 1‐α genes were sequenced from representative strains of each group to confirm the identifications. Fourteen Fusarium species were identified with F. graminearum (20%), F. verticillioides (18%), F. oxysporum (16%), F. subglutinans (13%) and F. proliferatum (11%) all present as more than 10% of the population. Fusarium acuminatum, F. avenaceum, F. concolor, F. crookwellense (F. cerealis), F. equiseti, F. semitectum, F. solani, F. sporotrichioides and F. venenatum, were all present at frequencies < 8%. Our results indicate that economically important Fusarium spp. may be isolated from numerous alternative hosts during the off season and that weeds and plant debris can serve as a reservoir of genetically diverse inoculum.