Characterization of yeasts isolated from red wine surface film

We have isolated 6 morphologically different axenic yeast cultures from the film surface of red wine. Based on morphological, physiological and biochemical characteristics we have identified the strains as follows: isolates1–4 are morphologically different strains of the anamorph basidiomycetous film-forming yeastCandida humicola (Daszewska) Diddens etLodder, syn.Apiotrichum humicola (Daszewska) von Arx Weijman. Isolates5 and6 belong to the genusSaccharomyces of the associated speciesS. cerevisiœ (isolate5 originallyS. bayanus, isolate6 S. capensis). These do not participate in the surface film formation.     

Molecular identification of yeasts from the order Trichosporonales causing superficial infections

BackgroundThe molecular reclassification of the order Trichosporonales placed the medically relevant Trichosporon species into three genera of the family Trichosporonaceae: Cutaneotrichosporon, Trichosporon, and Apiotrichum. From the clinical and epidemiological standpoint, it is important to identify any species of the family Trichosporonaceae because they present different antifungal susceptibility profiles. In Mexico, little is known about trichosporonosis etiology because the fungi are identified through phenotypic methods.AimsTo identify at a molecular level 12 yeast isolates morfologically compatible with Trichosporon, obtained from patients with superficial infections.MethodsThe yeast isolates were obtained from patients with white piedra, onychomycosis, and hand and foot dermatomycosis, and were identified morphologically and genotypically (sequencing of the IGS1 region and phylogenetic analysis using the Maximum Likelihood Method). The phylogenetic analysis included 40 yeast sequences from the order Trichosporonales and one from Cryptococcus neoformans as outgroup.ResultsBased on the molecular analysis, we identified three (25%) Trichosporon inkin isolates, two (16.7%) Trichosporon asteroides, two (16.7%) Cutaneotrichosporon mucoides, and one each (8.3%) of Trichosporon aquatile, Trichosporon asahii, Apiotrichum montevideense, Cutaneotrichosporon cutaneum, and Cutaneotrichosporon jirovecii.ConclusionsThe molecular characterization of the isolates showed a broad diversity of species within the order Trichosporonales, particularly among onychomycosis. It is essential to identify these yeasts at the species level to delve into their epidemiology.     

Yeasts from Tierra Del Fuego Province (Argentina): Biodiversity,Characterization and Bioprospection of Hydrolytic Enzymes

Abstract

Antarctic and sub-Antarctic regions are – with Polar Regions, mountains and the deep sea – the most extreme environments on Earth because of its low temperatures, dryness, high incidence of solar radiation and low nutrient availability. Nevertheless, microorganisms have successfully colonized these regions. In this study, culturable yeasts from soil samples collected from two different locations, a human-impacted area (Encerrada Bay) and a largely pristine and naturally vegetated area near Lago Escondido city (54°39′0″S, 67°46′48″W) from Tierra del Fuego province, Argentina were identified and characterized at different levels. They were characterized and classified as psychrotolerant and were considered as moderately halotolerant because of their ability to grow in the presence of 1.5?M of NaCl. Yeasts from phylum Ascomycota were affiliated to five genera: Candida, Yarrowia, Debaryomyces, Nadsonia, and Wickerhamiella, whereas from phylum Basidiomycota yeasts were affiliated to six genera: Naganishia, Rhodotorula, Leucosporidum, Tausonia, Cystofilobasidium, and Apiotrichum. Most of the yeasts demonstrated at least one extracellular enzymatic activity (mainly β-glucosidase, esterase, and protease activities). One isolate identified as Tausonia pullulans showed significant activity across the eight enzyme types tested. In light of these findings, Tierra del Fuego province could be considered as a cold environment with a potential source of cold-adapted yeasts producing industrially relevant cold-active enzymes.     

Distribution of Streptococcus troglodytae and Streptococcus dentirousetti in chimpanzee oral cavities

The aim of this study was to analyze the distribution and phenotypic properties of the indigenous streptococci in chimpanzee (Pan troglodytes) oral cavities. Eleven chimpanzees (aged from 9 to 44 years, mean ± SD, 26.9 ± 12.6 years) in the Primate Research Institute of Kyoto University were enrolled in this research and brushing bacterial samples collected from them. Streptococci were isolated from the oral cavities of all chimpanzees. The isolates (n = 46) were identified as thirteen species by 16S rRNA genes analysis. The predominant species was Streptococcus sanguinis of mitis streptococci from five chimpanzees (45%). Mutans streptococci were isolated from six chimpanzees (55%). The predominant species in the mutans streptococci were Streptococcus troglodytae from four chimpanzees (36%), this species having been proposed as a novel species by us, and Streptococcus dentirousetti from three chimpanzees (27%). Streptococcus mutans was isolated from one chimpanzee (9%). However, Streptococcus sobrinus, Streptococcus macacae and Streptococcus downei, which are indigenous to human and monkey (Macaca fasciclaris) oral habitats, were not isolated. Of the mutans streptococci, S. troglodytae, S. dentirousetti, and S. mutans possessed strong adherence activity to glass surface.     

Penicillium Mycobiota in Arctic Subglacial Ice

Fungi have been only rarely isolated from glacial ice in extremely cold polar regions and were in these cases considered as random, long-term preserved Aeolian deposits. Fungal presence has so far not been investigated in polar subglacial ice, a recently discovered extreme habitat reported to be inhabited exclusively by heterotrophic bacteria. In this study we report on the very high occurrence (up to 9000 CFU L⁻¹) and diversity of filamentous Penicillium spp. in the sediment-rich subglacial ice of three different polythermal Arctic glaciers (Svalbard, Norway). The dominant species was P. crustosum, representing on the average half of all isolated strains from all three glaciers. The other most frequently isolated species were P. bialowiezense, P. chrysogenum, P. thomii, P. solitum, P. palitans, P. echinulatum, P. polonicum, P. commune, P. discolor, P. expansum, and new Penicillium species (sp. 1). Twelve more Penicillium species were occasionally isolated. The fungi isolated produced consistent profiles of secondary metabolites, not different from the same Penicillium species from other habitats. This is the first report on the presence of large populations of Penicillium spp. in subglacial sediment-rich ice.     

Isolation,identification and characterization of Staphylococcus sp. from Indian ethnic fermented fish product

Staphylococci from Sheedal of Northeast India was isolated, identified and characterized. All the isolated staphylococci were found to be coagulase negative. Based on the rpoB gene sequences followed by analysis using NCBI-BLAST software, seven species of Staphylococcus namely, S. piscifermentans, S. condimenti, S. arlettae, S. sciuri, S. warneri, S. nepalensis and S. hominis were recognized. Phylogenetic analyses revealed three major cluster groups. All the seven Staphylococcus showed their NaCl tolerance from 2 to 8%. No species was able to grow at 55°C. Except S. arlettae and S. sciuri, all the isolated staphylococcal species exhibited growth at pH 4–8. No isolated species was able to ferment mannitol, sucrose and arabinose. All the species exhibited moderate to maximum proteolytic and lipolytic activities. All the seven species were found to be sensitive to the antibiotics, namely, erythromycin, norfloxacin, ampicillin, streptomycin and vancomycin, whereas all were resistant to co-trimoxazole. Only S. piscifermentans was found antagonist to Salmonella enterica, Escherichia coli and Bacillus subtilis, although the clear zone was minimal. All the staphylococcal species except S. arlettae and S. sciuri exhibited hydrophobicity ranging from 25 to 66%. The observed characteristics of isolated Staphylococci from Sheedal revealed their role in fish fermentation.     

Constant association of ophiostomatoid fungi with the bark beetle <Emphasis Type="Italic">Ips subelongatus</Emphasis> invading Japanese larch logs

Ceratocystiopsis, Ceratocystis, Grosmannia, and Ophiostoma species were isolated from Ips subelongatus and beetle-infested Japanese larch logs collected at several areas in central and northern Honshu Island, Japan, to determine constant associates of I. subelongatus. Ceratocystiopsis minuta, two species of Ceratocystis, three species of Grosmannia, and four species of Ophiostoma were isolated. Of the fungi isolated in the present study, G. laricis, O. brunneociliatum, and O. piceae were constant associates of the beetles. Ceratocystis fujiensis, Ceratocystiopsis minuta, and Ophiostoma sp. F were occasionally isolated with high frequencies of occurrence but were not consistent associates. Ceratocystis fujiensis was most often isolated as the leading fungal invasion in the sapwood of Japanese larch logs invaded by I. subelongatus, confirming that the fungus acts as a primary invader of sapwood in beetle-attacked logs. Contribution No. 221, Laboratory of Plant Parasitic Mycology, Graduate School of Life and Environmental Sciences, University of Tsukuba     

浙江雁荡山山脉叶栖酵母菌资源与物种多样性

对北雁荡山、中雁荡山、南雁荡山、西雁荡山和东雁荡山等广阔区域内的叶栖酵母菌资源进行考察,旨在弄清雁荡山山脉叶栖酵母菌资源和多样性。采用PDA平板对雁荡山地区的叶栖酵母菌资源进行分离和纯化,根据酵母菌培养物26S rDNA D1/D2序列和系统发育关系明确酵母菌的分类地位,分析该地区叶栖酵母菌群落的多样性、组成和分布情况。结果发现,分离自雁荡山山脉的1118株叶栖酵母菌可以被鉴定为56个已知类群和19个疑似新种,其中38种酵母菌在该地区比较常见。常见的酵母菌类群中,包括Derxomyces属5种,Sporobolomyces属3种,Bannoa、Bulleribasidium、Rhodosporidiobolus、Symmetrospora、Taphrina、Tilletiopsi和Udeniomyces属各2种,Bullera、Coniochaeta、Coniosporium、Cryptococcus、Elsinoe、Erythrobasidium、Fellozyma、Golubevia、Kockovaella、Kondoa、Leucosporidium、Moesziomyces、Oberwinklerozyma、Phyllozyma、Ruinenia和Saitozyma属各1种。根据常见酵母菌类群的相对频率,在该地区分离到的叶栖酵母菌中Derxomyces mrakii具有明显分布优势,并探讨了Derxomyces mrakii株间26S rDNA D1/D2序列的稳定性和变异幅度。根据雁荡山地区常见叶栖酵母菌组成和相似程度,可以推测地理上相近地区的酵母菌物种分布具有更大相似性。基于雁荡山山脉常见叶栖酵母菌种类和数量而绘制出的物种累积曲线,证明雁荡山地区绝大多数的叶栖酵母菌种类已经被分离到,研究结果可以真实反映该地区叶栖酵母菌资源和物种多样性的存在状况。     

Isolation and identification of endophytic and mycorrhizal fungi from seeds and roots of <Emphasis Type="Italic">Dendrobium</Emphasis> (Orchidaceae)

The seed germination of orchids under natural conditions requires association with mycorrhizal fungi. Dendrobium nobile and Dendrobium chrysanthum are threatened orchid species in China where they are considered medicinal plants. For conservation and application of Dendrobium using symbiosis technology, we isolated culturable endophytic and mycorrhizal fungi colonized in the protocorms and adult roots of two species plants and identified them by morphological and molecular analyses (5.8S and nrLSU). Of the 127 endophytic fungi isolated, 11 Rhizoctonia-like strains were identified as Tulasnellales (three strains from protocorms of D. nobile), Sebacinales (three strains from roots of D. nobile and two strains from protocorms of D. chrysanthum) and Cantharellales (three strains from roots of D. nobile), respectively. In addition, species of Xylaria, Fusarium, Trichoderma, Colletotrichum, Pestalotiopsis, and Phomopsis were the predominant non-mycorrhizal fungi isolated, and their probable ecological roles in the Dendrobium plants are discussed. These fungal resources will be of great importance for the large-scale cultivation of Dendrobium plants using symbiotic germination technology and for the screening of bioactive metabolites from them in the future.     

Soil Microfungi in Two Post-Mining Chronosequences with Different Vegetation Types

Soil microfungi were studied in the Sokolov (Czech Republic) post‐mining dumps afforested with Alnus glutinosa and in the Lusatian (Germany) post‐mining dumps afforested with Pinus sylvestris or P. nigra. Microfungi were isolated using the soil dilution plate method. Soil microfungi communities of two chronosequences were compared by species composition, frequency of species occurrence, and colony forming units of fungi (CFU‐counts). Differences in species occurrence were determined. More species of entomopathogenic microfungi were found from the Sokolov post‐mining area in comparison with the Cottbus post‐mining area. Absidia glauca, A. cylindrospora, Penicillium glabrum, and P. janthinellum were the most frequently isolated species from the Cottbus post‐mining area, while A. glauca, Geomyces pannorum, and Trichoderma koningii predominated at the Sokolov post‐mining area.     

Characterization of clinical isolates of pathogenic <Emphasis Type="Italic">Nocardia</Emphasis> strains and related actinomycetes in Thailand from 1996 to 2003

In Thailand from 1996 to 2003, 171 strains of pathogenic aerobic actinomycetes from clinical specimens were isolated. Of those strains, 134 were mycolic acid containing actinomycetes, including 96 strains of Nocardia species. Others included 10 strains of Gordonia, 14 strains of Rhodococcus, and 22 strains of Mycobacterium. One strain each of the genera Tsukamurella and Corynebacterium were also isolated. Also identified were 27 strains of non-mycolic acid containing actinomycetes. Our identification studies of 96 strains of Nocardia species showed that significant pathogens in Thailand were N. beijingensis (18 strains), N. cyriacigeorgica (13 strains), and N. farcinica (34 strains); the most prevalent species was N. farcinica (35.4%). We also isolated four strains of N. asiatica, five strains of N. asteroides sensu stricto, four strains of N. nova, seven strains of N. otitidiscaviarum, eight strains of N. transvalensis, and two strains of N. pseudobrasiliensis.     

Isolation for bacteria and fungi for the hydrolysis of phthalate and terephthalate esters

Bacterial and fungal strains were isolated from enrichment cultures using diethylphthalate, diethylterephthalate, or ethylene glycol dibenzoate as sole carbon sources.Aureobacterium, Flavobacterium, andMicrococcus species were isolated from diethylphthalate enrichments;Rhodococcus andXanthomonas species were isolated from diethylterephthalate enrichments;Rhodococcus andFusarium species were isolated from ethylene glycol dibenzoate enrichments.     

Isolation and characterization of arsenite-oxidizing bacteria from arsenic-contaminated soils in Thailand

Two hundred and eighty-eight arsenic-resistant bacteria were isolated by an enrichment culture method from a total of 69 arsenic-contaminated soil-samples collected from Dantchaeng district in Suphanburi province (47 samples), and from Ron Phiboon district in Nakhon Sri Thammarat province (22 samples), in Central and Southern Thailand, respectively. Twenty-four of the 288 isolated arsenic-resistant bacteria were found to be arsenite-oxidizing bacteria. On the basis of their morphological, cultural, physiological, biochemical and chemotaxonomic characteristics, and supported by phylogenetic analysis based upon their 16S rRNA gene sequences, they were divided into five groups, within the genera Acinetobacter, Flavobacterium, Pseudomonas, Sinorhizobium and Sphingomonas, respectively. Within genera, phylogenetic analysis using the 16S rRNA gene sequences suggested that they were comprised of at least ten species, five isolates being closely related to known bacteria (Acinetobacter calcoaceticus NCCB 22016^T, Pseudomonas plecoglossicida FPC951^T, Ps. knackmussii B13^T, Sinorhizobium morelense Lc04^T, and Sphingomonas subterranea IFO16086^T). The other five proposed species are likely to be new species closely related to Flavobacterium johnsoniae, Sinorhizobium morelense, Acinetobacter calcoaceticus and Pseudomonas plecoglossicida, but this awaits further characterization for confirmation of the taxonomic status. No overlap in isolated species or strains was observed between the two sites. The strain distribution and characterization are described.     

Population dynamics of bacteria associated with different strains of the pine wood nematode Bursaphelenchus xylophilus after inoculation in maritime pine (Pinus pinaster)

For a long time it was thought that Bursaphelenchus xylophilus was the only agent of the pine wilt disease. Recently, it was discovered that there are bacteria associated with the nematodes that contribute to the pathogenesis of this disease, mainly through the release of toxins that promote the death of the pines. Among the species most commonly found, are bacteria belonging to the Bacillus, Pantoea, Pseudomonas and Xanthomonas genera.The main objective of this work was to study the effect of inoculation of maritime pine (Pinus pinaster) with four different nematode isolates, in the bacterial population of nematodes and trees, at different stages of disease progression. The monitoring of progression of disease symptoms was also recorded. Also, the identification of bacteria isolated from the xylem of trees and the surface of nematodes was performed by classical identification methods, by the API20E identification system and by sequencing of bacterial DNA.The results showed that for the symptoms progression, the most striking difference was observed for the pines inoculated with the avirulent isolate, C14-5, which led to a slower and less severe aggravation of symptoms than in pines inoculated with the virulent isolates. In general, it was found that bacterial population, inside the tree, increased with disease progression. A superior bacterial quantity was isolated from pines inoculated with the nematode isolates HF and 20, and, comparatively, few bacteria were isolated from pines inoculated with the avirulent isolate. The identification system API20E was insufficient in the identification of bacterial species; Enterobacter cloacae species was identified in 79% of the isolated bacterial colonies and seven of these colonies could not be identified by this method. Molecular identification methods, through bacterial DNA sequencing, allowed a more reliable identification: eleven different bacterial species within the Bacillus, Citrobacter, Enterobacter, Escherichia, Klebsiella, Paenibacillus, Pantoea and Terribacillus genera were identified. General bacterial diversity increased with the progression of the disease. Bacillus spp. were predominant at the earlier stage of disease progression and Klebsiella oxytoca at the later stages. Furthermore, bacterial species isolated from the surface of nematodes were similar to those isolated from the xylem of pines.In the present work new bacterial species were identified which have never been reported before in this type of study and may be associated with their geographical origin (Portugal). P. pinaster, the pine species used in this study, was different from those commonly grown in Japan and China. Furthermore, it was the first time that bacteria were isolated and identified from an avirulent pine wood nematode isolate.     

Conidiation and carbohydrate composition in some Candida and Torulopsis species

The genus Candida Berkhout (including Torulopsis auct.) is restricted to the anamorphs of Saccharomycetales (asci-forming yeasts). Species with heterobasidiomycetous relationships are either transferred to Apiotrichum when the cells are at least partly formed sympodially and when the hydrolyzates of intact cells contain xylose, or to Rhodotorula when the cells are formed basipetally (enteroblastic, phialidic) and the cells do not contain xylose.     

A study of yeasts during the delignification and fungal transformation of wood into cattle feed in Chilean rain forest

In Chilean rain forest the presence and distribution of yeasts were studied during the fungal transformation of wood, producing a partially delignified material used as cattle feed (huempe). A total of 68 yeast species was reported, including a noticeable proportions of basidiomycetous and xylose-assimilating strains, the latter related to the increased availability of this sugar in the decayed wood. The results of the mathematic comparison of taxonomic data of 327 strains showed a delimitation of sharp clusters corresponding to different species, including several new species described from this substrate, and genera, as Rhodotorula and Cryptococcus.At the initial stages of wood decay, characterized by the presence of Candida species, yeast diversity was low. The highest values were found in the medium stage of decay, used as feed, with an important presence of the genera Apiotrichum, Rhodotorula, Cryptococcus and Schizoblastosporion. After establishing a non-linear ordination from Euclidean distances between samples, based on yeast abundances, a relationship was observed between the populations developing at the different stages of wood decay by Ganoderma applanatum. A succession of yeast species was demonstrated during wood degradation by this white-rot fungus.     

Antimicrobial Activity and Biodiversity of Endophytic Fungi in <Emphasis Type="Italic">Dendrobium</Emphasis><Emphasis Type="Italic">devonianum</Emphasis> and <Emphasis Type="Italic">Dendrobium thyrsiflorum</Emphasis> from Vietman

Endophytic fungi are rich in orchids and have great impacts on their host plants. 53 endophytes (30 isolates from Dendrobium devonianum and 23 endophytic fungi from D. thyrsiflorum) were isolated, respectively, from roots and stems of Dendrobium species. All the fungi were identified by way of morphological and/or molecular biological methods. 30 endophytic fungi in D. devonianum were categorized into 11 taxa and 23 fungal endophytes in D. thyrsiflorum were grouped into 11 genera, respectively. Fusarium was the dominant species of the two Dendrobium species in common. Antimicrobial activity of ethanol extract of fermentation broth of these fungi was explored using agar diffusion test. 10 endophytic fungi in D. devonianum and 11 in D. thyrsiflorum exhibited antimicrobial activity against at least one pathogenic bacterium or fungus among 6 pathogenic microbes (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus). Out of the fungal endophytes isolated from D. devonianum and D. thyrsiflorum, Phoma displayed strong inhibitory activity (inhibition zones in diameter >20 mm) against pathogens. Epicoccum nigrum from D. thyrsiflorum exhibited antibacterial activity even stronger than ampicillin sodium. Fusarium isolated from the two Dendrobium species was effective against the pathogenic bacterial as well as fungal pathogens. The study reinforced the assumption that endophytic fungi isolated from different Dendrobium species could be of potential antibacterial or antifungal resource.     

Mycoflora and mycotoxins in Brazilian black pepper,white pepper and Brazil nuts

A wide range of field and storage fungi were isolated from black pepper, white pepper and Brazil nut kernels from Amazonia. A total of 42 species were isolated from both peppers. Aspergillus flavus and A. niger were isolated more frequently from black than from white pepper. Other potential mycotoxigenic species isolated included: A. ochraceus, A. tamarii, A. versicolor, Emericella nidulans and Chaetomium globosum, Penicillium brevicompactum, P. citrinum, P. islandicum and P. glabrum. Species isolated from pepper for the first time were Acrogenospora sphaerocephala, Cylindrocarpon lichenicola, Lacellinopsis sacchari, Microascus cinereus, Petriella setifera and Sporormiella minima. Seventeen species were isolated from Brazil nut kernels. A. flavus was the dominant species followed by A. niger. P. citrinum and P. glabrum were the only penicillia isolated. Species isolated for the first time included Acremonium curvulum, Cunninghamella elegans, Exophiala sp., Fusarium oxysporum, Pseudoallescheria boydii, Rhizopusoryzae, Scopulariopsis sp., Thielavia terricola and Trichoderma citrinoviride. Considerably more metabolites were detected from black than white pepper in qualitative analyses. Chaetocin, penitrem A, and xanthocillin were identified only from black pepper, and tenuazonic acid was identified from both black and white pepper. Aflatoxin G₂, chaetoglobosin C, and spinulosin were identified from poor quality brazil nuts. Aflatoxin B₁ and B₂ were also only detected in poor quality brazil nuts at concentrations of 27.1 g kg^–1 and 2.1 g kg^–1 respectively (total 29.2 g kg^–1).This revised version was published online in October 2005 with corrections to the Cover Date.     

Evaluation of the enteric microflora of captive whooping cranes (Grus americana) and sandhill cranes (Grus canadensis)

The enteric flora of captive whooping cranes (Grus americana) and sandhill cranes (Grus canadensis) has not been well described, despite its potential importance in the understanding of both the normal condition of the intestinal physiology of these animals and the altered colonization within disease states in these birds. Nineteen whooping cranes and 23 sandhill cranes housed currently at the Calgary Zoo or its affiliated Devonian Wildlife Conservation Centre (DWCC) in Calgary, Alberta were sampled from October 2004–February 2005 by collecting aerobic and anaerobic cloacal swabs from each bird. There were seven major groupings of bacteria isolated from both species of crane. Gram‐positive cocci, coliforms, and gram‐negative bacilli were the most prevalent types of bacteria isolated for both crane species, with Escherichia coli, Enterococcus faecalis, and Streptococcus Group D, not Enterococcus the bacterial species isolated most commonly. There was a significant difference in the average number of isolates per individual between the two crane species but no differences between age or gender categories within crane species. Campylobacter sp. were isolated from five whooping cranes. The potential zoonotic pathogen Campylobacter jejuni was isolated from one whooping crane and C. upsaliensis was isolated from a second. Three other isolates were unspeciated members of the Campylobacter genus and likely belong to a species undescribed previously. The evaluation of the enteric cloacal flora of whooping cranes and sandhill cranes illustrates that differences exist between these two closely related crane species, and highlights the potential implications these differences may have for current practices involving captive wildlife. Zoo Biol 0:1–13, 2007. © 2007 Wiley‐Liss, Inc.     

Sequence and Phylogenetic Analysis of SSU rRNA Gene of Five Microsporidia

The complete small subunit rRNA (SSU rRNA) gene sequences of five microsporidia including Nosema heliothidis, and four novel microsporidia isolated from Pieris rapae, Phyllobrotica armta, Hemerophila atrilineata, and Bombyx mori, respectively, were obtained by PCR amplification, cloning, and sequencing. Two phylogenetic trees based on SSU rRNA sequences had been constructed by using Neighbor-Joining of Phylip software and UPGMA of MEGA4.0 software. The taxonomic status of four novel microsporidia was determined by analysis of phylogenetic relationship, length, G+C content, identity, and divergence of the SSU rRNA sequences. The results showed that the microsporidia isolated from Pieris rapae, Phyllobrotica armta, and Hemerophila atrilineata have close phylogenetic relationship with the Nosema, while another microsporidium isolated from Bombyx mori is closely related to the Endoreticulatus. So, we temporarily classify three novel species of microsporidia to genus Nosema, as Nosema sp. PR, Nosema sp. PA, Nosema sp. HA. Another is temporarily classified into genus Endoreticulatus, as Endoreticulatus sp. Zhenjiang. The result indicated as well that it is feasible and valuable to elucidate phylogenetic relationships and taxonomic status of microsporidian species by analyzing information from SSU rRNA sequences of microsporidia.