Studies on the genus Paecilomyces in China V. Taifanglania gen. nov for some monophialidic species

A rare monophialidic fungus, Taifanglania hechuanensis gen. & sp. nov., was isolated from soil oil the banks of Jialin River, Hechuan, Chongqing City during a survey of soil-borne filamentous fungi from different phytogeographical areas in China. It is described and illustrated in this paper. A further eight monophialidic species of Paecilomyces are transferred to the genus. Diagnosis features of the new genus are white, grey, straw yellow or brown to black colonies on Czapek agar. Conidiophores are always absent or simple. Phialides are solitary, consisting of a cylindrical or ellipsoidal swollen basal portion, tapering into a thin neck, directly arising on vegetative hyphae or prophialides, sometimes consisting of a whorl of 2 to 3 phialides oil simple conidiophores. Conidia arc one-celled, hyaline, smooth-walled, subglobose, ellipsoidal or fusiform, having or no the connective between conidia and being thermotolerant. The new species is characterized by pale yellow to grey-yellow colonies, solitary phialides with ail ellipsoidal or fusiform basal portion that arise directly from the vegetative hyphae, big conidia (3.1-)3.9-8.7 x ( 1.7-)2.1-4.7(-5.1) mu m with the connective, and thermotolerant growth. A molecular study based oil the nucleotidic sequences of the SSU rDNA and ITS regions support the status of T. hechuanensis as a new species and Taifanglania as a new genus.

Isolation of culturable yeasts from market wines and evaluation of the 5 center dot 8S-ITS rDNA sequence analysis for identification purposes

Aims: To test the possibility that wines available in the marketplace may contain culturable yeasts and to evaluate the 5.8S-ITS rDNA sequence analysis as adequate means for the identification of isolates. Methods and Results: As a case study, typical Greek wines were surveyed. Sequence analysis of the 5.8S-ITS rDNA was tested for its robustness in species or strain identification. Sixteen isolates could be assigned into the species Brettanomyces bruxellensis, Saccharomyces cerevisiae and Rhodotorula pinicola, whereas four isolates could not be safely identified. B. bruxellensis was the dominant species present in house wines, while non-Saccharomyces sp. were viable in aged wines of high alcohol content. Conclusions: Yeast population depends on postfermentation procedures or storage conditions. Although 5.8S-ITS rDNA sequence analysis is generally a rapid method to identify wine yeast isolates at the species level, or even below that, it may not be sufficient for some genera. Significance and Impact of the Study: This is the first report to show that commercial wines may possess diverse and potentially harmful yeast populations. The knowledge of yeasts able to reside in this niche environment is essential towards integrated quality assurance programmes. For selected species, the 5.8S-ITS rDNA sequence analysis is a rapid and accurate means.

Analysis of 16S rDNA sequences from pathogenic Leptospira serovars and use of single nucleotide polymorphisms for rapid speciation by D-HPLC

Leptospira have a worldwide distribution and include important zoonotic pathogens yet diagnosis and differentiation still tend to rely on traditional bacteriological and serological approaches. In this study a 1.3 kb fragment of the rrs gene (16S rDNA) was sequenced from a panel of 22 control strains, representing serovars within the pathogenic species Leptospira interrogans, Leptospira borgpetersenii, and Leptospira kirschneri, to identify single nucleotide polymorphisms (SNPs). These were identified in the 5' variable region of the 16S sequence and a 181 bp PCR fragment encompassing this region was used for speciation by Denaturing High Performance Liquid Chromatography (D-HPLC). This method was applied to eleven additional species, representing pathogenic, non-pathogenic and intermediate species and was demonstrated to rapidly differentiate all but 2 of the non-pathogenic Leptospira species. The method was applied successfully to infected tissues from field samples proving its value for diagnosing leptospiral infections found in animals in the UK. Crown Copyright (C) 2010 Published by Elsevier Ltd. All rights reserved.

Distribution of mosquitoes in the South East of Argentina and first report on the analysis based on 18S rDNA and COI sequences

Although Mar del Plata is the most important city on the Atlantic coast of Argentina, mosquitoes inhabiting such area are almost uncharacterized. To increase our knowledge in their distribution, we sampled specimens of natural populations. After the morphological identification based on taxonomic keys, sequences of DNA from small ribosomal subunit (18S rDNA) and cytochrome c oxidase I (COI) genes were obtained from native species and the phylogenetic analysis of these sequences were done. Fourteen species from the genera Uranotaenia, Culex, Ochlerotatus and Psorophora were found and identified. Our 18S rDNA and COI-based analysis indicates the relationships among groups at the supra-species level in concordance with mosquito taxonomy. The introduction and spread of vectors and diseases carried by them are not known in Mar del Plata, but some of the species found in this study were reported as pathogen vectors.

A method for merging nadir-sounding climate records, with an application to the global-mean stratospheric temperature data sets from SSU and AMSU

A method is proposed for merging different nadir-sounding climate data records using measurements from high-resolution limb sounders to provide a transfer function between the different nadir measurements. The two nadir-sounding records need not be overlapping so long as the limb-sounding record bridges between them. The method is applied to global-mean stratospheric temperatures from the NOAA Climate Data Records based on the Stratospheric Sounding Unit (SSU) and the Advanced Microwave Sounding Unit-A (AMSU), extending the SSU record forward in time to yield a continuous data set from 1979 to present, and providing a simple framework for extending the SSU record into the future using AMSU. SSU and AMSU are bridged using temperature measurements from the Michelson Interferometer for Passive Atmospheric Sounding (MIPAS), which is of high enough vertical resolution to accurately represent the weighting functions of both SSU and AMSU. For this application, a purely statistical approach is not viable since the different nadir channels are not sufficiently linearly independent, statistically speaking. The near-global-mean linear temperature trends for extended SSU for 1980–2012 are −0.63 ± 0.13, −0.71 ± 0.15 and −0.80 ± 0.17 K decade−1 (95 % confidence) for channels 1, 2 and 3, respectively. The extended SSU temperature changes are in good agreement with those from the Microwave Limb Sounder (MLS) on the Aura satellite, with both exhibiting a cooling trend of ~ 0.6 ± 0.3 K decade−1 in the upper stratosphere from 2004 to 2012. The extended SSU record is found to be in agreement with high-top coupled atmosphere–ocean models over the 1980–2012 period, including the continued cooling over the first decade of the 21st century.

Temperature trends derived from Stratospheric Sounding Unit radiances: the effect of increasing CO2 on the weighting function

Stratospheric Sounding Units (SSU) on the NOAA operational satellites have been the main source of near global temperature trend data above the lower stratosphere. They have been used extensively for comparison with model-derived trends. The SSU senses in the 15 micron band of CO2 and hence the weighting function is sensitive to changes in CO2 concentrations. The impact of this change in weighting function has been ignored in all recent trend analyses. We show that the apparent trends in global mean brightness temperature due to the change in weighting function vary from about -0.4 K/decade to 0.4 K/decade depending on the altitude sensed by the different SSU channels. For some channels, this apparent trend is of a similar size to the trend deduced from SSU data but ignoring the change in weighting function. In the mid-stratosphere, the revised trends are now significantly more negative and in better agreement with model-calculated trends.

Use of an artificial root to examine the influence of 8-hydroxyquinoline on soil microbial activity and bacterial community structure

Invasive plant species have been shown to alter the microbial community composition of the soils they invade and it is suggested that this below-ground perturbation of potential pathogens, decomposers or symbionts may feedback positively to allow invasive success. Whether these perturbations are mediated through specific components of root exudation are not understood. We focussed on 8-hydroxyquinoline, a putative allelochemical of Centaurea diffusa (diffuse knapweed) and used an artificial root system to differentiate the effects of 8-hydroxyquinoline against a background of total rhizodeposition as mimicked through supply of a synthetic exudate solution. In soil proximal (0-10 cm) to the artificial root, synthetic exudates had a highly significant (P < 0.001) influence on dehydrogenase, fluorescein diacetate hydrolysis and urease activity. in addition, 8-hydroxyquinoline was significant (p = 0.003) as a main effect on dehydrogenase activity and interacted with synthetic exudates to affect urease activity (p = 0.09). Hierarchical cluster analysis of 16S rDNA-based DGGE band patterns also identified a primary affect of synthetic exudates and a secondary affect of 8-hydroxyquinoline on bacterial community structure. Thus, we show that the artificial rhizosphere produced by the synthetic exudates was the predominant effect, but, that the influence of the 8-hydroxyquinoline signal on the activity and structure of soil microbial communities could also be detected. (C) 2009 Elsevier Ltd. All rights reserved.

Stratospheric temperature and radiative forcing response to 11-year solar cycle changes in irradiance and ozone

The 11-yr solar cycle temperature response to spectrally resolved solar irradiance changes and associated ozone changes is calculated using a fixed dynamical heating (FDH) model. Imposed ozone changes are from satellite observations, in contrast to some earlier studies. A maximum of 1.6 K is found in the equatorial upper stratosphere and a secondary maximum of 0.4 K in the equatorial lower stratosphere, forming a double peak in the vertical. The upper maximum is primarily due to the irradiance changes while the lower maximum is due to the imposed ozone changes. The results compare well with analyses using the 40-yr ECMWF Re-Analysis (ERA-40) and NCEP/NCAR datasets. The equatorial lower stratospheric structure is reproduced even though, by definition, the FDH calculations exclude dynamically driven temperature changes, suggesting an important role for an indirect dynamical effect through ozone redistribution. The results also suggest that differences between the Stratospheric Sounding Unit (SSU)/Microwave Sounding Unit (MSU) and ERA-40 estimates of the solar cycle signal can be explained by the poor vertical resolution of the SSU/MSU measurements. The adjusted radiative forcing of climate change is also investigated. The forcing due to irradiance changes was 0.14 W m−2, which is only 78% of the value obtained by employing the standard method of simple scaling of the total solar irradiance (TSI) change. The difference arises because much of the change in TSI is at wavelengths where ozone absorbs strongly. The forcing due to the ozone change was only 0.004 W m−2 owing to strong compensation between negative shortwave and positive longwave forcings.

Ultrastructure of Buddenbrockia allmani n sp (Myxozoa, Malacosporea), a parasite of Lophopus crystallinus (Bryozoa, Phylactolaemata)

Development of a new species of malacosporean myxozoan (Buddenbrockia allmani n. sp.) in the bryozoan Lophopus crystallinus is described. Early stages, represented by isolated cells or small groups, were observed in the host's body wall or body cavity. Multiplication and rearrangement of cells gave an outer cell layer around a central mass. The outer cells made contact by filopodia and established adherens junctions. Sporoplasmosomes were a notable feature of early stages, but these were lost in subsequent development. Typical malacosporean sacs were formed from these groups by attachment of the inner (luminal) cells by a basal lamina to the outer layer (mural cells). Division of luminal cells gave rise to a population of cells that was liberated into the lumen of the sac. Mitotic spindles in open mitosis and prophase stages of meiosis were observed in luminal cells. Centrioles were absent. Detached luminal cells assembled to form spores with four polar capsules and several valve cells surrounding two sporoplasms with secondary cells. Restoration of sporoplasmosomes occurred in primary sporoplasms. A second type of sac was observed with highly irregular mural cells and stellate luminal cells. A radially striated layer and dense granules in the polar capsule wall, and previous data on 18 rDNA sequences enabled assignment of the species to the genus Buddenbrockia, while specific diagnosis relied on the rDNA data and on sac shape and size.

Are Cape floral clades the same age? Contemporaneous origins of two lineages in the genistoids s.l. (Fabaceae)

The hypothesis that the elements of the modern species-rich flora of the Cape Floristic Region (CFR), South Africa, originated more or less simultaneously at the Miocene/Pliocene boundary, in response to the development of a mediterranean climate, has been challenged by numerous molecular dating estimates of Cape floral clades. These studies reveal a more gradual emergence, with the oldest clades originating in the Eocene, but others appearing later, some as recently as the Pliocene. That there are factors which might affect the dates recovered, such as choice of calibration point, analysis method, sampling density and the delimitation of Cape floral clades, suggests a need for further critical evaluation of the age estimates presented to date. In this study, the dates of origin of two Cape floral clades (the legume Crotalarieae p.p. and Podalyrieae) are estimated, constrained by a shared calibration point in a single analysis using an rDNA ITS phylogeny in which 633 taxa are sampled. The results indicate that these two clades arose contemporaneously 44-46 mya, not at the Miocene/Pliocene boundary as had been previously supposed. The contemporaneous origin of these Cape floral clades suggests that additional more inclusive analyses are needed before rejecting the hypothesis that a. single environmental trigger explains the establishment of Cape floral clades. (c) 2007 Elsevier Inc. All rights reserved.

Diversity, phylogeny and distribution of bean rhizobia in salt-affected soils of North-West Morocco

Different molecular methods: BOX-PCR fingerprinting, R-FLP-PCR and sequencing of the 16S rDNA as well as the symbiotic genes nodC and nifH, were used to study the genetic diversity within a collection of nodulating bean rhizobia isolated from five soils of North-West Morocco. BOX fingerprints analysis of 241 isolates revealed 19 different BOX profiles. According to the PFLP-PCR and sequencing of 16S rDNA carried out on 45 representative isolates, 5 genotypes were obtained corresponding to the species Rhizobium etli, R. tropici, R. gallicum, R. leguminosarum and Sinorhizobium meliloti. The most abundant species were R. etli and R. tropici (61% and 24%, respectively). A high intraspecific diversity was observed among the R. etli isolates, while the R. tropici group was homogeneous. Most of the rhizobia studied belong to species known to nodulate common bean, while 2 species were unconventional microsymbionts: R. leguminosarum biovar viciae and S. meliloti. Our results, especially the nodulation promiscuity of common bean and the relation between the predominance of some species of rhizobia in particular soils and the salt content of these soils, indicate that there is a real need for a better understanding of the distribution of common bean rhizobia species in the soils of Morocco before any inoculation attempt.

Diversity and systematics of the Malacosporea (Myxozoa)

Myxozoans belonging to the recently described class Malacosporea parasitize freshwater bryozoans during at least part of their life cycle. There are at present only two species described in this class: Buddenbrockia plumatellae and Tetracapsuloides bryosalmonae. The former can exist as vermiform and sac-like stages in bryozoan hosts. The latter, in addition to forming sac-like stages in bryozoans, is the causative agent of salmonid proliferative kidney disease (PKD). We undertook molecular and ultrastructural investigations of new malacosporean material to further resolve malacosporean diversity and systematics. Phylogenetic analyses of 18S rDNA sequences provided evidence for two new putative species belonging to the genus Buddenbrockia, revealing a two-fold increase in the diversity of malacosporeans known to date. One new malacosporean is a vermiform parasite infecting the bryozoan Fredericella sultana and the other occurs as sac-like stages in the rare bryozoan, Lophopus crystallinus. Both bryozoans represent new hosts for the genus Buddenbrockia. Our results have established that the malacosporean which infected F. sultana was not a vermiform stage of T. bryosalmonae, although it was collected from a site endemic for PKD. Ultrastructural investigation of new material of B. plumatellae revealed the presence of numerous external tubes associated with developing polar capsules, confirming that the absence of external tubes should no longer be considered as a character of the class Malacosporea.

Hespellia stercorisuis gen. nov., sp nov and Hespellia porcina sp nov., isolated from swine manure storage pits

Four Gram-positive-staining, strictly anaerobic, non-spore-forming, rod-shaped organisms were isolated from a pig manure storage pit. Comparative 16S rRNA gene sequence analysis revealed that the isolates belonged to two related but distinct groups. Sequence analysis showed that the two groups of isolates were highly related to each other (approx. 97% 16S rRNA gene sequence similarity), forming a distinct cluster within the Clostridium coccoides suprageneric rDNA grouping. Biochemical and physiological studies confirmed the division of the isolates into two related, albeit distinct, groups. Based on both phenotypic and phylogenetic evidence, it is proposed that the unidentified rod-shaped isolates from pig manure should be classified in a novel genus, Hespellia gen. nov., as Hespellia stercorisuis sp. nov. and Hespellia porcina sp. nov. The type species of the novel genus is H. stercorisuis (type strain, PC18(T) = NRRL B-23456(T) = CCUG 46279(T) = ATCC BAA-677(T)) and the type strain of H. porcina is PC80(T) (= NRRL B-23458(T) = ATCC BAA-674(T)).

Molecular identification and anti-pathogenic activities of putative probiotic bacteria isolated from faeces of healthy elderly individuals

One hundred and nine lactic acid bacterial strains (56 bifidobacteria-like and 53 lactobacilli-like) were isolated from faecal samples donated by healthy elderly individuals (>65 years old). Isolates were identified to species level by phenotypic analysis (by API) and by 16S rDNA sequencing. Eleven species of Lactobacillus and six species of Bifidobacterium were identified. The most frequently isolated lactobacillus was L. fermentum and the most frequently isolated bifidobacterium was closely related to B. infantis by 16S rDNA sequence alignment. The isolates were characterized for their antimicrobial activity against Clostridium difficile, enteropathogenic Escherichia coli (EPEC), verocytotoxigenic E. coli (VTEC) and Campylobacter jejuni. The lactobacilli displayed variations in their antimicrobial activity with few strains showing inhibitory activity against all pathogens. The bifidobacteria displayed higher levels of inhibitory activity against C. jejuni and Cl. difficile than against the E. coli strains. Keywords: Lactobacillus, Bifidobacterium, elderly, gastrointestinal microbiota, inhibition, Clostridium difficile, enteropathogenic Escherichia coli (EPEC), verocytotoxigenic E. coli (VTEC), Campylobacter jejuni.