Human leptospirosis caused by a new, antigenically unique Leptospira associated with a Rattus species reservoir in the Peruvian Amazon

As part of a prospective study of leptospirosis and biodiversity of Leptospira in the Peruvian Amazon, a new Leptospira species was isolated from humans with acute febrile illness. Field trapping identified this leptospire in peridomestic rats (Rattus norvegicus, six isolates; R. rattus, two isolates) obtained in urban, peri-urban, and rural areas of the Iquitos region. Novelty of this species was proven by serological typing, 16S ribosomal RNA gene sequencing, pulsed-field gel electrophoresis, and DNA-DNA hybridization analysis. We have named this species "Leptospira licerasiae" serovar Varillal, and have determined that it is phylogenetically related to, but genetically distinct from, other intermediate Leptospira such as L. fainei and L. inadai. The type strain is serovar Varillal strain VAR 010(T), which has been deposited into internationally accessible culture collections. By microscopic agglutination test, "Leptospira licerasiae" serovar Varillal was antigenically distinct from all known serogroups of Leptospira except for low level cross-reaction with rabbit anti-L. fainei serovar Hurstbridge at a titer of 1:100. LipL32, although not detectable by PCR, was detectable in "Leptospira licerasiae" serovar Varillal by both Southern blot hybridization and Western immunoblot, although on immunoblot, the predicted protein was significantly smaller (27 kDa) than that of L. interrogans and L. kirschneri (32 kDa). Isolation was rare from humans (2/45 Leptospira isolates from 881 febrile patients sampled), but high titers of MAT antibodies against "Leptospira licerasiae" serovar Varillal were common (30%) among patients fulfilling serological criteria for acute leptospirosis in the Iquitos region, and uncommon (7%) elsewhere in Peru. This new leptospiral species reflects Amazonian biodiversity and has evolved to become an important cause of leptospirosis in the Peruvian Amazon.     

Fatty acid composition of Leptospira lipids as a taxonomic criterion

The fatty-acid composition of microbial cells in 17 pathogenic and saprophytic Leptospira strains, comprising 14 serovars and 10 serogroups, has been studied. The strains under investigation have proved to fall into 3 groups differing by this characteristic. The group of saprophytic strains is characterized by a comparatively high level of myristic acid and, consequently, by the ratio of saturated and unsaturated fatty acids with 14 carbon atoms approaching 1:1; besides, it is also characterized by a lower, in comparison with the pathogenic Leptospira strains belonging to the serogroups Icterohaemorrhagiae, Canicola, Ballum has a higher level of unsaturated fatty acids. The second group of pathogenic Leptospira strains including the serogroups Grippotyphosa, Hebdomadis, Pomona, Tarassovi, Pyrogenes, Australia has been found to occupy an intermediate position between the first group of pathogenic Leptospira strains and the group of saprophytic ones. As the difference in the content of myristic acid in pathogenic and saprophytic Leptospira strains is a stable characteristic, it can be used for the differentiation of these strains. The present investigation has revealed that the distribution of the main fatty acids in Leptospira phospholipids is similar to their distribution in Leptospira neutral lipids with the exception of unsaturated fatty acid with 14 carbon atoms, occurring mainly in phospholipids.     

The functioning of a drained natural focus of leptospirosis in a nonchernozem area

The drainage of a natural focus of leptospirosis of the flood plain-swamp type, carried out over the period of 15 years, has led to changes in the species structure of small mammals and to an increase in the number of Leptospira-carrying species. Nevertheless, like before drainage, the prevalent species and the main carrier of leptospires is still the root vole (Microtus oeconomus Pallas). The intensity of the epizootic process among small mammals at the final stage of drainage has proved to be similar to that at the initial stage. At the same time the cultivated fields created on the drained territory of the natural focus abound, by the time of harvesting, with small mammals among whom the intensive epizootic process takes place.     

湖北四湖流域景观结构变化对汛期湖泊水位影响的模拟分析

借助于水文学对不同下垫面状态的产流模式研究的成果,建立了计算四湖流域两个调蓄湖泊汛期洪峰水位的模型。模拟分析的结果表明,在4种同样的降雨条件下,景观结构变化使长湖汛期日水位平均增加0．0831—1．3223m,最大增加0．1579—1．8583m,使洪湖汛期日水位平均增加1.0810一2．3008m,最大增加1．4101—2．4864m。证明景观结构变化是引起洪涝灾害加剧的重要原因且景观结构变化对湖泊洪峰水位的影响存在一种累加效应。景观结构的调整也是控制洪涝灾害的重要途径。     

Testaceans (Testacea, Protozoa) of Taiga Soils in Western Siberia (Surgut Polesye)

We studied population structure of testaceans of automorphic and hydromorphic soils in the middle taiga of Western Siberia. Rich testacean fauna has been revealed. The fauna representation was comparable to the studied soils of Ciscaucasian Russia. The bryophilic group was the most typical for the region, since its representatives inhabited the predominating hydromorphic soils of bogs. The typical species included Amphitrema flavum, Heleopera petricola, Placocista spinosa, Trigonopyxis arcula, Centropyxis laevigata, Nebela tincta, Arcella catinus, as well as other moss forms. Flood plain proved to be the richest by testacean species variability among the studied habitats. The set of species here included both representatives of watershed and typical aquatic forms present due to the flood conditions of a given area. The highest quantitative indices of the testacean complexes were revealed in the forest area soils. Quadrulella quadrigera Deflandre, 1936 found in a flood plain area has been revealed for the first time in Russia.     

Flood plain wetland fisheries of India: with special reference to impact of climate change

Floodplain wetlands of India are biologically rich sensitive ecosystem that support unique aquatic biodiversity and play vital role in providing livelihood and nutritional security to a large section of the population of the country. Besides contributing to the environmental sustainability through Carbon sequestration, flood plain wetlands also serve as source for harvesting flood and rain water. Covering around 5.5 Lakh ha area, the flood plain wetlands of India are one of the major sources for fish production of the country, offering vast potential for capture as well as culture based fisheries. Degradation and shrinkage of the floodplain wetlands have been recorded due to several natural and anthropogenic reasons. In addition to that, the change in the climatic condition may have a far more devastating impact on these natural resources. Substantial change in climate with increasing temperature trend (0.60 °C during last 112 years) as well as changing pattern and intensity of rainfall have been reported in India. It is projected that extreme climate changes may have profound impact on wetlands, mediated through several direct or indirect pathways. Attention is urgently required at different levels for conservation and revamping of these resources along with coping up and mitigation strategies to address the impending challenges. However, there is dearth of scientific information specific to the flood plain wetlands of India. This paper reviews the present status and importance of the flood plain wetlands of India with special reference to impact of climate change along with coping-up and mitigation measures.     

Cultivation of Leptospira in a liquid culture medium with lysed rabbit blood

A new method of the preparation of liquid culture medium for the cultivation of Leptospira has been developed on the basis of multiple freezing and thawing of defibrinated rabbit blood. A higher productivity of the new medium in comparison with the medium used in common practice has been established. The medium having a new composition is suitable for cultivation of Leptospira and the accumulation of biomass. The advantage of the medium made is in addition to high growth properties, the its economic efficiency and availability for practical laboratories.     

挠力河流域平原区湿地景观完整性评价

探讨一种景观完整性评价方法对湿地管理和保护非常重要.利用遥感和GIS技术,选择10个指标构建景观完整性指数,对挠力河流域平原区开展景观完整性评价.结果显示,1950年挠力河流域洪泛平原中的湿地景观完整性远好于2005年;人类活动影响越强的区域湿地景观完整性越差;2005年仅河岸带湿地景观比其它区域具有较好的景观完整性.由于景观完整性指数包涵了必要的景观结构和功能完整性特征信息,对评价景观尺度生态完整性,诊断生态受损区域,保护、恢复和管理湿地具有重要作用.     

Floating vegetation mats on a floodplain billabong in the Northern Territory of Australia

The floristics and physical structure of a floating mat of vegetation in the Finniss River area of the Northern Territory are described. During the dry season floating mats contain green productive vegetation when the adjacent flood plains are dry and many of the plants are withered. At this time insects are more abundant on the mats than on the adjacent flood plain. Some flood plain vertebrates appear to concentrate their activity on floating mats during the dry season. During the period 1963 to 1978 the area of floating mat in the Finniss River study area was greatly reduced, probably as a result of high densities of feral water buffalo (Bubalis bubalus).(Present address: c/o 38 Hardy Terrace, East Ivanhoe, Victoria 3079).(Present address: c/o 38 Hardy Terrace, East Ivanhoe, Victoria 3079).     

Comparative study of the phospholipase activity of pathogenic and saprophytic Leptospira cultured on a serum-lecithin agar

The phospholipase activity of leptospires cultivated on serum-lecithin agar has been studied. Two zones of changes in the medium have been found to appear around the colonies of saprophytic Leptospira strains: transparent (5.25 +/- 2.09 mm wide) and turbid (6.90 +/- +/- 1.46 mm wide), which is linked with the production of phospholipases A and C. Only a single clear zone is formed around the colonies of pathogenic strains due to the production of phospholipase A. At the same time virulent Leptospira strains show greater phospholipase activity (the zones are 6.0 +/- 1.2 mm wide) than avirulent strains (the zones are 1.6 +/- +/- 0.04 mm wide).     

Random insertional mutagenesis of Leptospira interrogans, the agent of leptospirosis, using a mariner transposon

The recent availability of the complete genome sequences of Leptospira interrogans, the agent of leptospirosis, has allowed the identification of several putative virulence factors. However, to our knowledge, attempts to carry out gene transfer in pathogenic Leptospira spp. have failed so far. In this study, we show that the Himar1 mariner transposon permits random mutagenesis in the pathogen L. interrogans. We have identified genes that have been interrupted by Himar1 insertion in 35 L. interrogans mutants. This approach of transposon mutagenesis will be useful for understanding the spirochetal physiology and the pathogenic mechanisms of Leptospira, which remain largely unknown.     

Identification and partial characterization of a novel hemolysin from Leptospira interrogans serovar lai

It has been suggested that leptospiral hemolysins are important in the virulence and pathogenesis of leptospirosis. We have isolated an Escherichia coli clone carrying the 7.8kb DNA insert from a genomic library of Leptospira interrogans serovar lai by plaque hybridization using a sequence derived from the sphingomyelinase C gene (sphA) of L. borgpetersenii. The clone showed a clear beta-hemolytic zone on sheep blood agar and high hemolytic activities on both human and sheep erythrocytes in liquid assays. The clone carried at least two genes responsible for the hemolytic activities, encoded by two open reading frames of 1662 and 816 nucleotides, which are named sphH and hap-1 (hemolysis associated protein-1), respectively. The SphH showed 75% homology to the SphA at the amino acid level, and the Hap-1 showed no significant homology in major databases. Interestingly, however, E. coli cells harboring sphH did not show sphingomyelinase or phospholipase activities. Moreover, SphH-mediated hemolysis was osmotically protected by polyethylene glycol 5000, suggesting that the hemolysis is likely to be caused by pore formation on the membrane. The SphH was successfully expressed in E. coli as a histidine (His)-SphH fusion protein. Both sphH and hap-1 were highly conserved among the Leptospira species, except for the absence of sphH in non-pathogenic L. biflexa serovar patoc. We concluded that the SphH is a novel hemolysin of a pathogenic Leptospira species, which may be a putative pore-forming protein.     

Conservation of the S10-spc-alpha locus within otherwise highly plastic genomes provides phylogenetic insight into the genus Leptospira

S10-spc-alpha is a 17.5 kb cluster of 32 genes encoding ribosomal proteins. This locus has an unusual composition and organization in Leptospira interrogans. We demonstrate the highly conserved nature of this region among diverse Leptospira and show its utility as a phylogenetically informative region. Comparative analyses were performed by PCR using primer sets covering the whole locus. Correctly sized fragments were obtained by PCR from all L. interrogans strains tested for each primer set indicating that this locus is well conserved in this species. Few differences were detected in amplification profiles between different pathogenic species, indicating that the S10-spc-alpha locus is conserved among pathogenic Leptospira. In contrast, PCR analysis of this locus using DNA from saprophytic Leptospira species and species with an intermediate pathogenic capacity generated varied results. Sequence alignment of the S10-spc-alpha locus from two pathogenic species, L. interrogans and L. borgpetersenii, with the corresponding locus from the saprophyte L. biflexa serovar Patoc showed that genetic organization of this locus is well conserved within Leptospira. Multilocus sequence typing (MLST) of four conserved regions resulted in the construction of well-defined phylogenetic trees that help resolve questions about the interrelationships of pathogenic Leptospira. Based on the results of secY sequence analysis, we found that reliable species identification of pathogenic Leptospira is possible by comparative analysis of a 245 bp region commonly used as a target for diagnostic PCR for leptospirosis. Comparative analysis of Leptospira strains revealed that strain H6 previously classified as L. inadai actually belongs to the pathogenic species L. interrogans and that L. meyeri strain ICF phylogenetically co-localized with the pathogenic clusters. These findings demonstrate that the S10-spc-alpha locus is highly conserved throughout the genus and may be more useful in comparing evolution of the genus than loci studied previously.     

Electrophoretic study of the outer membrane proteins of leptospirae

The outer membranes of pathogenic and saprophytic leptospires have been isolated. The spectrum of outer membrane proteins in three saprophytic and one pathogenic Leptospira strains has been studied by means of electrophoresis in polyacrylamide gel. In Leptospira strains VGNKI-6 (pathogenic) and G-80 (saprophytic) identical proteins, as well as proteins similar in their Rf value, have been detected. The possibility of using strain G-80 for the development of leptospiral vaccine against serovars having common surface antigens with this strain has been suggested.     

The impact of flooding regime on the soil seed bank of flood‐meadows

Abstract. We assessed the significance of flooding for the floristic composition of seed banks in flood‐meadows of the northern valley of the Upper Rhine. We compared three hydrological compartments of the alluvial plain, consisting of the regularly flooded land between the river and low summer dykes (functional flood‐plain), the occasionally flooded land between summer dykes and high winter dykes (hybrid floodplain) and the land behind the winter dykes, which is now only submerged by ascending groundwater (fossil flood‐plain). Due to their different flooding regime, the three compartments should differ with respect to the prevailing conditions of diaspore input. The seed density of soil samples increased with the duration of flooding in the three compartments, while species richness and the proportion of species not occurring in the vegetation was constant. The increase in seed density can be largely attributed to an increase of disturbance indicators, which are present in the above‐ground vegetation and capable of forming a long‐term persistent seed bank. No effects of flooding on the composition of seed banks in the three flood‐plain compartments were found. The differences in seed bank composition can be largely explained by corresponding differences in above‐ground vegetation and former and present‐day meadow management. Seeds of species absent from above‐ground vegetation can be attributed to the local species pool present in the immediate vicinity of the study plots. We discuss consequences of the results for the restoration of species‐rich flood‐plain meadows.     

内蒙古高原典型草原区河漫滩湿地植物群落退化表征

草原区河流河漫滩草甸是生物多样性表现最充分和生物生产力最高的地段, 但由于过度放牧利用, 绝大部分草甸处于退化状态。该文以锡林河流域中游的河漫滩草甸为研究对象, 比较分析了围封保育湿地与放牧退化湿地的群落组成、地上生物量, 以及共有植物种的植株高度、节间长、叶长、叶宽, 土壤含水量、容重, 群落地下根量及根的分布, 土壤微生物生物量碳、氮的变化。结果表明: 1)放牧使得湿地植物群落优势种发生变化, 原有湿生植物逐渐向旱生化转变, 同时地上及地下生物量明显降低。2)退化湿地的植物呈现显著小型化现象。3)放牧退化湿地的土壤含水量较围封保育湿地低, 其垂直分布及地下根的垂直分布也发生变化。在低河漫滩, 土壤水分随土层的增加而增加, 根量也趋于深层化。但在高河漫滩湿地, 土壤含水量接近典型草原, 根未出现深层化分布趋势。4)放牧践踏引起土壤容重和土壤紧实度增加。5)放牧使得低河漫滩湿地土壤微生物生物量增加, 而在过渡区及高河漫滩湿地, 放牧使得土壤微生物生物量碳、氮含量显著降低。     

基于SWAT模型的北京平原区森林景观格局对雨洪减缓的影响研究

景观格局及其变化是影响地表径流及空间分布的重要因素,也是管理城市雨洪、治理生态环境的途径。以北京市平原区百万亩造林项目为背景,选取第一轮造林时间（2011、2013、2015、2017年）的遥感数据,以三环、五环、六环路为界,将平原区划分为核心区、中心区、近郊区、远郊区四个区域,分别分析整体及分区林地景观格局的变化。运用SWAT模型模拟不同格局对不同区域暴雨径流的影响,探究影响径流变化的主导林地景观指数。研究发现：1）平原区林地面积呈增长趋势,增幅为12.96％。除核心区和中心区的部分地区外,林地斑块更聚集、形状更复杂。2）百万亩造林工程对平原区雨洪风险的减缓有积极作用,但受城市建设及人类活动的影响,不同区域造林对径流的减缓强度与建设扩张的影响强度大小关系略有不同。核心区、中心区造林对径流的减缓强度大于建设扩张对径流的副作用,径流总量减小,减少比例为1.98％、4.29％。而近郊区、远郊区相反,径流总量变大,增大比例为0.09％与6.82％,其中远郊区建设扩张的强度是影响平原区径流风险的主导原因。3）不同区域雨洪减缓的关键性林地景观指数不同。核心区径流的主导林地因子为斑块数量与形状因子,中心区为面积因子,近郊区为聚合度因子和形状因子,远郊区为面积与聚合度因子。主导林地因子的大小调控将更高效的减缓雨洪风险。本研究通过SWAT模拟分析得到第一轮造林期间平原区森林格局与径流特征的空间变化及各区域径流减缓的关键性林地因子,为第二轮造林提高雨洪减缓功效提供参考。     

Management history determines gene flow in a prominent invader

Invasive plants pose substantial threats to protected areas globally. Although management can limit impacts, spread and reinvasion from neighbouring areas into protected areas are a major and an on‐going problem for land managers. However, identifying the main sources of propagules and the dimensions of invasion pathways is challenging. This study used population genetic markers [inter simple sequence repeats (ISSRs) and amplified fragment length polymorphisms (AFLPs)] to infer the source(s) of re‐colonization and dispersal patterns for a typical invader of riparian and terrestrial habitats (Lantana camara) along the Sabie‐Sand catchment, one of the most important river systems flowing into and across South Africa’s flagship protected area, the Kruger National Park (KNP). Results indicate that populations located along the lower reaches of the Sabie and Sand tributaries harboured substantially higher genetic diversity than those in the upper Sabie catchment. Bayesian assignments indicated that the upper Sabie tributary contributed far fewer propagules than the Sand tributary to the lower Sabie River. Current invasion patterns are due to a combination of a major flood event in 2000 and differences in the degree to which the upstream reaches were managed after the flooding. The major flood of 2000 effectively cleared lantana from the riparian areas. However, whereas on‐going management efforts against riparian species in the KNP have been effective, rendering the upper Sabie relatively clear of lantana, only a small part of the Sand tributary falls under jurisdiction of the KNP and has received consistent management attention. The reinvasion of the lower Sabie in the KNP was therefore almost entirely by propagules from the Sand tributary. The study highlights the important role that molecular tools can play in determining dispersal dynamics and directing invasive species management. For invasive plant species that invade both riparian habitats and landscapes away from rivers in protected areas, such as lantana, management must focus on all major sources of propagules to limit reinvasion.     

Pollination and phenology of flowers in the canopy of two contrasting rain forest types in Amazonia,Colombia

The main objective of this investigation was to study the pollination characteristics of two types of Amazonian rain forest at plant community level. Seasonally inundated forest was compared with upland (tierra firme) forest. The study focused on plant species in the canopy. The pollination spectra show that in both forests most canopy trees and lianas are pollinated by small bees, large bees, butterflies or by small, relatively unspecialized insects. In the upland forest small bees are the most important pollinators (32% of all species of trees and lianas are pollinated by them), whereas large bees are predominant in the floodplain (22%). Other pollinators, like hummingbirds, bats, moths, and beetles are less common (>10%), but always somewhat more important in the flood plain than in the upland forest. Bees are the most common pollinators of epiphytes. In the flood plain forest, flies are also important as epiphyte pollinators (19%), whereas in the upland forest hummingbirds pollinate more epiphytes. The phenological patterns are quite similar in both the upland and the flood plain. We found a peak in flowering in the transition period between the wet and the dry season. Flowering activity was lowest during the wet season. Differentiation in sexual systems was correlated with life form. Dioecy and monoecy were found mostly among tree species. Most species of all life forms though were hermaphroditic. No difference with respect to the relative importance of sexual systems was found between the two forest types.     

Molecular typing of Leptospira spp. based on putative O-antigen polymerase gene (wzy), the benefit over 16S rRNA gene sequence

Molecular typing of leptospiral strains based on variation within putative O-antigen polymerase gene (wzy) was determined among reference strains and those isolated from patients. Using the PCR primers designed from the flanking gene of wzy derived from Leptospira interrogans serovar Copenhageni, all L. interrogans serovars as well as human and rodent leptospiral isolates from Thailand could be amplified. The size of PCR product ranged from 1 to 1.5 kb. The limitation of these primer pairs was the inability to amplify those strains whose sequences differ in the region of the primers, these included Leptospira biflexa (serovar Patoc), Leptospira borgpetersenii (serovar Tarassovi) and Leptospira kirschneri (serovar Bim, Bulgarica, Butembo). Notably, amplification was not limited to L. interrogans as demonstrated by the amplification of some strains from L. kirschneri, Leptospira meyeri, Leptospira noguchii, Leptospira santarosai, L. borgpetersenii and Leptospira weilii. The phylogenetic tree of wzy sequence, inferred by posterior probability of the Bayesian, enabled the categorization of leptospiral serovars into seven genetically related group, of which its differentiation power was better than that of the more highly conserved 16S rRNA gene, which is used extensively for genotyping.