Short-Term In Vitro Culture and Molecular Analysis of the Microsporidian, Enterocytozoon bieneusi

ABSTRACT. The microsporidium, Enterocytozoon bieneusi , causes a severe, debilitating, chronic diarrhea in patients with the acquired immunodeficiency syndrome. Specific diagnosis of intestinal microsporidiosis, especially due to Enterocytozoon , is difficult and there is no known therapy that can completely eradicate this parasite. Preliminary studies indicate that a short term (about 6 months) in vitro culture of this parasite yielding low numbers of spores, may be established by inoculating human lung fibroblasts and/or monkey kidney cell cultures with duodenal aspirates and or biopsy from infected patients. The cultures may subsequently be used for the isolation and molecular analysis of parasite DNA.     

Antigenic Differences Among Epimastigotes,Amastigotes and Trypomastigotes of Trypanosoma cruzi*

Antigenic differences were demonstrated among trypomastigotes, amastigotes, and epimastigotes of Trypanosoma cruzi by the indirect fluorescent antibody method. Tests using cross-absorbed sera were included in the study.     

Crystals in woody stems

Scanning electron microscopy has been used to examine the location and form of crystals in the stems of woody perennials. The results are discussed in relation to the effects of physical conditions and "impurities" upon crystal form. Infrared spectroscopy and X-ray diffraction have been used to establish the chemical identity of the crystals in a number of species which were chosen so as to provide a wide variation in form. Those from Populus deltoides and Polyalthia sp. consisted of calcium carbonate; those from ten other species consisted of hydrated calcium oxalate. The infrared spectra of the crystals from Dracaena fragrans, Terminalia balerica, Goniothalamus sp. and Combretum verticillatum were strongly indicative of calcium oxalate monohydrate, but in the other cases the degree of hydration of the oxalate was uncertain.     

The Comparative Wood-destroying Ability and Preservative Tolerance of Monocaryotic and Dicaryotic Mycelia of Lenzites trabea (Pers.) Fr. and Poria vaillantii (DC ex Fr.) Cke.

Comparison by soil-block tests on several timbers of the wood-destroyingabilities of dicaryotic cultures of both Poria vaillantii andLenzites trabea with those of monocaryotic cultures either derivedfrom, or contributing to, the formation of the dicaryon, indicatedthat monocaryotic cultures of P. vaillantii were generally moredestructive than related dicaryotic cultures, whereas dicaryonsof L. trabea tended to be slightly more destructive than relatedmonocaryons. The tolerance of L. trabea monocaryons to coppersulphate and to sodium arsenate in nutrient agar showed someindication of being higher than that of related dicaryons, whilstthe tolerance of P. vaillantii monocaryons to a copperchrome-arsenatepreservative in soil-block tests also appeared to be higherthan that of related dicaryons. It is concluded that, althoughneither wood-destroying ability nor preservative tolerance wasgreatly affected by nuclear condition, the differences shownmay be of importance in conjunction with the monocaryotizationof dicaryons by toxic agents during laboratory decay tests.     

Genetic uniformity characterizes the invasive spread of water hyacinth (Eichhornia crassipes), a clonal aquatic plant

Aquatic plant invasions are often associated with long‐distance dispersal of vegetative propagules and prolific clonal reproduction. These reproductive features combined with genetic bottlenecks have the potential to severely limit genetic diversity in invasive populations. To investigate this question we conducted a global scale population genetic survey using amplified fragment length polymorphism markers of the world’s most successful aquatic plant invader –Eichhornia crassipes (water hyacinth). We sampled 1140 ramets from 54 populations from the native (South America) and introduced range (Asia, Africa, Europe, North America, Central America and the Caribbean). Although we detected 49 clones, introduced populations exhibited very low genetic diversity and little differentiation compared with those from the native range, and ～80% of introduced populations were composed of a single clone. A widespread clone (‘W’) detected in two Peruvian populations accounted for 70.9% of the individuals sampled and dominated in 74.5% of the introduced populations. However, samples from Bangladesh and Indonesia were composed of different genotypes, implicating multiple introductions to the introduced range. Nine of 47 introduced populations contained clonal diversity suggesting that sexual recruitment occurs in some invasive sites where environmental conditions favour seedling establishment. The global patterns of genetic diversity in E. crassipes likely result from severe genetic bottlenecks during colonization and prolific clonal propagation. The prevalence of the ‘W’ genotype throughout the invasive range may be explained by stochastic sampling, or possibly because of pre‐adaptation of the ‘W’ genotype to tolerate low temperatures.     

上海乡土水生植物资源及其在水生态恢复与水景观建设中的应用潜力

水生植物作为水生生态系统的主体, 对发挥水生生态系统的自维持、自循环功能有重要作用。研究通过相关资料的查阅, 建立上海地区乡土水生植物名录, 并对其科属组成、区系特征、生活型、生长型等进行统计分析。结果表明上海地区乡土水生植物共计35 科83 属160 种(含变种), 单属科、单种属的比例较高, 均达65%以上; 植物区系组成丰富、成分复杂, 以热带成分占优势, 达64.6%; 生活型以挺水植物为主, 沉水植物次之, 浮水植物最少; 生长型类型丰富, 以草本型、禾草型居多, 20 种生长型可进一步归为表征相似生态学特征和功能地位的6 个生长型组。在水生态恢复与水景观建设中, 仅有68.8%的景观水体有水生植物应用,且应用种类在2 种以下的占79.2%。乡土水生植物应用不足, 一半以上为观赏性强的外来物种, 应用频率较高的为挺水植物, 对具有良好净化效果的沉水植物重视不够。因此, 在水生态恢复与水景观建设中, 建议加强乡土水生植物资源的繁育栽培, 在充分利用乡土水生植物资源配置群落的基础上, 根据水质的富营养及基底状况, 通过不同生长型组水生植物的应用, 构建沉水-浮水-挺水植物群落复合体, 并通过近自然型护岸的营造, 形成水生-湿生复合生态系统。运用植被工程学的原理和方法, 采用生态浮岛、生态沉岛等技术营造水生植被, 将强人工化的水景观建成具生命的水生生态系统。       

Phylogenetic relationships among marsupial frog genera (Anura: Hylidae: Hemiphractinae) based on evidence from morphology and natural history

We reassess the phylogenetic relationships of genera of hemiphractine hylid frogs (Marsupial Treefrogs) and discuss the evolution of several distinctive characters within this group using parsimony analysis. Fifty-one morphological and life-history characters were sampled from two species of Cryptobatrachus , three species of Flectonotus , 17 species of Gastrotheca , all five species of Hemiphractus , and one species of Stefania as the ingroup and three hyline, one phyllomedusine, and one pelodryadine species as outgroups. Our results support the mon-ophyly of Flectonotus, Cryptobatrachus , and Hemiphractus. Gastrotheca is paraphyletic with respect to Hemiphractus , dorsal pouches were lost in the ancestor of Hemiphractus. Direct development is a synapomorphy for Hemiphractinae and tadpoles were regained independently several times. These results stand in stark contrast to the prevailing paradigm regarding marsupial frog relationships.     

Host range expansion of an introduced insect pest through multiple colonizations of specialized clones

Asexuality confers demographic advantages to invasive taxa, but generally limits adaptive potential for colonizing of new habitats. Therefore, pre-existing adaptations and habitat tolerance are essential in the success of asexual invaders. We investigated these key factors of invasiveness by assessing reproductive modes and host-plant adaptations in the pea aphid, Acyrthosiphon pisum, a pest recently introduced into Chile. The pea aphid encompasses lineages differing in their reproductive mode, ranging from obligatory cyclical parthenogenesis to fully asexual reproduction. This species also shows variation in host use, with distinct biotypes specialized on different species of legumes as well as more polyphagous populations. In central Chile, microsatellite genotyping of pea aphids sampled on five crops and wild legumes revealed three main clonal genotypes, which showed striking associations with particular host plants rather than sampling locations. Phenotypic analyses confirmed their strong host specialization and demonstrated parthenogenesis as their sole reproductive mode. The genetic relatedness of these clonal genotypes with corresponding host-specialized populations from the Old World indicated that each clone descended from a particular Eurasian biotype, which involved at least three successful introduction events followed by spread on different crops. This study illustrates that multiple introductions of highly specialized clones, rather than local evolution in resource use and/or selection of generalist genotypes, can explain the demographic success of a strictly asexual invader.     

Structural and functional differentiation of two clinally distributed glucosephosphate isomerase allelic isozymes from the teleost Fundulus heteroclitus

The teleost Fundulus heteroclitus (L.) possesses two loci, Gpi-A and Gpi-B, for the glycolytic enzyme, glucose-phosphate isomerase (GPI; D- glucose-6-phosphate ketol-isomerase; E.C. 5.3.1.9). The Gpi-B locus is polymorphic in Fundulus, with two common alleles, Gpi-Bb and Gpi-Bc, distributed in a clinal manner in populations along the east coast of North America. Since this clinal distribution is strongly correlated with a temperature gradient, we asked whether the GPI-B2 allozymes were functionally adapted to the thermal environment in which a given phenotype predominated. The two major GPI-B2 allozymes were purified to homogeneity and were characterized as to molecular weight, isoelectric pH, thermal denaturation, and kinetic parameters. Both GPI-Bb2 and GPI- Bc2 allozymes have molecular masses of 110 kD, and they have isoelectric pHs of 6.4 and 6.6, respectively. The GPI-Bb2 allozyme was more stable to thermal denaturation than was the GPI-Bc2 enzyme. Kinetic properties of the allelic isozymes were investigated both as a function of pH and as a function of temperature. At 25 degrees C, over the pH range considered, there were no significant differences between allozymes, either in Km for fructose-6-phosphate or in Ki for 6- phosphogluconate, but apparent Vmax values differed between pH 7.5 and pH 8.5. All steady-state kinetic parameters showed strong temperature dependence, but the allozymes differed only in the Ki for 6- phosphogluconate at temperatures greater than 30 degrees C. On the basis of the observed structural and functional differences alluded to above, the hypothesis that the major allelic isozymes of the Gpi-B locus were functionally equivalent was rejected. However, it is not yet known whether these structural and functional differences have any significance at higher levels of biological organization.