Phylogenetic and Functional Diversity Within Toluene-Degrading,Sulphate-Reducing Consortia Enriched from a Contaminated Aquifer

Three toluene-degrading microbial consortia were enriched under sulphate-reducing conditions from different zones of a benzene, toluene, ethylbenzene and xylenes (BTEX) plume of two connected contaminated aquifers. Two cultures were obtained from a weakly contaminated zone of the lower aquifer, while one culture originated from the highly contaminated upper aquifer. We hypothesised that the different habitat characteristics are reflected by distinct degrader populations. Degradation of toluene with concomitant production of sulphide was demonstrated in laboratory microcosms and the enrichment cultures were phylogenetically characterised. The benzylsuccinate synthase alpha-subunit (bssA) marker gene, encoding the enzyme initiating anaerobic toluene degradation, was targeted to characterise the catabolic diversity within the enrichment cultures. It was shown that the hydrogeochemical parameters in the different zones of the plume determined the microbial composition of the enrichment cultures. Both enrichment cultures from the weakly contaminated zone were of a very similar composition, dominated by Deltaproteobacteria with the Desulfobulbaceae (a Desulfopila-related phylotype) as key players. Two different bssA sequence types were found, which were both affiliated to genes from sulphate-reducing Deltaproteobacteria. In contrast, the enrichment culture from the highly contaminated zone was dominated by Clostridia with a Desulfosporosinus-related phylotype as presumed key player. A distinct bssA sequence type with high similarity to other recently detected sequences from clostridial toluene degraders was dominant in this culture. This work contributes to our understanding of the niche partitioning between degrader populations in distinct compartments of BTEX-contaminated aquifers.     

Geographical trends in the yolk carotenoid composition of the pied flycatcher (Ficedula hypoleuca)

Carotenoids in the egg yolks of birds are considered to be important antioxidants and immune stimulants during the rapid growth of embryos. Yolk carotenoid composition is strongly affected by the carotenoid composition of the female??s diet at the time of egg formation. Spatial and temporal differences in carotenoid availability may thus be reflected in yolk concentrations. To assess whether yolk carotenoid concentrations or carotenoid profiles show any large-scale geographical trends or differences among habitats, we collected yolk samples from 16 European populations of the pied flycatcher, Ficedula hypoleuca. We found that the concentrations and proportions of lutein and some other xanthophylls in the egg yolks decreased from Central Europe northwards. The most southern population (which is also the one found at the highest altitude) also showed relatively low carotenoid levels. Concentrations of ??-carotene and zeaxanthin did not show any obvious geographical gradients. Egg yolks also contained proportionally more lutein and other xanthophylls in deciduous than in mixed or coniferous habitats. We suggest that latitudinal gradients in lutein and xanthophylls reflect the lower availability of lutein-rich food items in the northern F. hypoleuca populations and in montane southern populations, which start egg-laying earlier relative to tree phenology than the Central European populations. Similarly, among-habitat variation is likely to reflect the better availability of lutein-rich food in deciduous forests. Our study is the first to indicate that the concentration and profile of yolk carotenoids may show large-scale spatial variation among populations in different parts of the species?? geographical range. Further studies are needed to test the fitness effects of this geographical variation.     

Lead Content in Pot Marigold (<Emphasis Type="Italic">Calendula officinalis</Emphasis> L.) Inflorescences and Leaves: Impact of Precipitations and Vicinity of Motorway

Trace metal contamination is a major environmental and health problem virtually in all countries. The present study was aimed to estimate the lead content of pot marigold (Calendula officinalis L.) inflorescences and leaves collected from a nonpolluted test field. The lead content in dry pot marigold inflorescences was 9.34 ± 0.79 μg/g, in dry leaves 11.57 ± 0.47 μg/g, and in soil 0.649 ± 0.012 μg/g. The distance of pot marigold collection beds (30–220 m from the motorway) had no effect on lead content. There was a strong positive correlation between the amount of precipitations and lead content of pot marigold leaves but not inflorescences indicating the soil as primarily the source of increased lead content. In conclusion, no effect of motorway vicinity was found for pot marigold inflorescences or leaves lead content; however, as a precaution, it is not recommended to collect the plants during or just after showers.     

Nitric oxide and nitrosative stress tolerance in yeast

The opportunistic human fungal pathogen Candida albicans encounters diverse environmental stresses when it is in contact with its host. When colonizing and invading human tissues, C. albicans is exposed to ROS (reactive oxygen species) and RNIs (reactive nitrogen intermediates). ROS and RNIs are generated in the first line of host defence by phagocytic cells such as macrophages and neutrophils. In order to escape these host-induced oxidative and nitrosative stresses, C. albicans has developed various detoxification mechanisms. One such mechanism is the detoxification of NO (nitric oxide) to nitrate by the flavohaemoglobin enzyme CaYhb1. Members of the haemoglobin superfamily are highly conserved and are found in archaea, eukaryotes and bacteria. Flavohaemoglobins have a dioxygenase activity [NOD (NO dioxygenase domain)] and contain three domains: a globin domain, an FAD-binding domain and an NAD(P)-binding domain. In the present paper, we examine the nitrosative stress response in three fungal models: the pathogenic yeast C. albicans, the benign budding yeast Saccharomyces cerevisiae and the benign fission yeast Schizosaccharomyces pombe. We compare their enzymatic and non-enzymatic NO and RNI detoxification mechanisms and summarize fungal responses to nitrosative stress.     

Archaeal Population Dynamics during Sequential Reduction Processes in Rice Field Soil

The population dynamics of Archaea after flooding of an Italian rice field soil were studied over 17 days. Anoxically incubated rice field soil slurries exhibited a typical sequence of reduction processes characterized by reduction of nitrate, Fe³⁺, and sulfate prior to the initiation of methane production. Archaeal population dynamics were followed using a dual approach involving molecular sequence retrieval and fingerprinting of small-subunit (SSU) rRNA genes. We retrieved archaeal sequences from four clone libraries (30 each) constructed for different time points (days 0, 1, 8, and 17) after flooding of the soil. The clones could be assigned to known methanogens (i.e., Methanosarcinaceae, Methanosaetaceae, Methanomicrobiaceae, and Methanobacteriaceae) and to novel euryarchaeotal (rice clusters I, II, and III) and crenarchaeotal (rice clusters IV and VI) lineages previously detected in anoxic rice field soil and on rice roots (R. Grosskopf, S. Stubner, and W. Liesack, Appl. Environ. Microbiol. 64:4983–4989, 1998). During the initiation of methanogenesis (days 0 to 17), we detected significant changes in the frequency of individual clones, especially of those affiliated with the Methanosaetaceae and Methanobacteriaceae. However, these findings could not be confirmed by terminal restriction fragment length polymorphism (T-RFLP) analysis of SSU rDNA amplicons. Most likely, the fluctuations in sequence composition of clone libraries resulted from cloning bias. Clonal SSU rRNA gene sequences were used to define operational taxonomic units (OTUs) for T-RFLP analysis, which were distinguished by group-specific TaqI restriction sites. Sequence analysis showed a high degree of conservation of TaqI restriction sites within the different archaeal lineages present in Italian rice field soil. Direct T-RFLP analysis of archaeal populations in rice field soil slurries revealed the presence of all archaeal lineages detected by cloning with a predominance of terminal restriction fragments characteristic of rice cluster I (389 bp), Methanosaetaceae (280 bp), and Methanosarcinaceae/rice cluster VI (182 bp). In general, the relative gene frequency of most detected OTUs remained rather constant over time during the first 17 days after flooding of the soil. Most minor OTUs (e.g., Methanomicrobiaceae and rice cluster III) and Methanosaetaceae did not change in relative frequency. Rice cluster I (37 to 30%) and to a lesser extent rice cluster IV as well as Methanobacteriaceae decreased over time. Only the relative abundance of Methanosarcinaceae (182 bp) increased, roughly doubling from 15 to 29% of total archaeal gene frequency within the first 11 days, which was positively correlated to the dynamics of acetate and formate concentrations. Our results indicate that a functionally dynamic ecosystem, a rice field soil after flooding, was linked to a relatively stable archaeal community structure.     

Revisiting the 1991 algal bloom in shelf waters off Argentina: Azadinium luciferelloides sp. nov. (Amphidomataceae,Dinophyceae) as the causative species in a diverse community of other amphidomataceans

Species of the marine dinophycean genera Azadinium and Amphidoma (Amphidomataceae) mostly attract attention because of their production potential of the lipophilic polyether phycotoxin azaspiracid (AZA). The genus Azadinium probably has a very wide geographical distribution. Blooms of Azadinium from the continental shelf off Argentina have been observed back in the early 1990, but were just recently published, and the causative species, identified at that time as Azadinium cf. spinosum, could not unequivocally be determined. Here we retrospectively analyzed old archived samples of one of the South Atlantic Azadinium bloom from 1991 with electron microscopy. It turned out that the dominant nanoplanktonic dinophycean species in fact represent a new species which we describe here based on the morphology. Azadinium luciferelloides sp. nov. is a small (approximately 9–14 μm cell length) thecate dinoflagellate with the dominant plate pattern of the genus (Po, X, 4′, 3a, 6″, 6C, 5S, 6″′, 2″″), and with a small antapical spine. Azadinium luciferelloides differed from all other described species of Azadinium by the position of the ventral pore, which was located on the right ventral side in a notch of an otherwise symmetric pore plate. In addition, we recorded and documented the presence of other similar sized species of the Amphidomataceae in the samples. Our finding of Az. spinosum, Az. dalianense, Az. dexteroporum, and Amphidoma languida are the first record for the South Atlantic and thus describe an important range extension of these species. The diversity and importance of the Amphidomataceae for South Atlantic spring bloom plankton is now known and taxonomically documented, but cultures and/or analysis of AZA in field samples of the area are needed to clarify the AZA production potential of the local species and populations in order to finally evaluate the risk potential of AZA for AZA shellfish contamination in the Southwestern Atlantic region.     

Development and application of visual traps for monitoring and control of populations ofRhagoletis cerasi L.

The authors — members of the OILB working group on genetic control of the Europan cherry fruit fly,Rhagoletis cerasi L. — describe in this second report the development of a standard trap for the target species and its applications in the framework of the genetic control procedures in progress. The trap has increased the attractiveness for the flies about 100 times compared with the traditional bait traps and has proven its effectiveness in recent suppression programs in the field.

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Résumé Les auteurs, membres du groupe de travail OILB sur la lutte génétique contre la mouche européenne des cerises,R. cerasi, décrivent la mise au point d'un piège standard pour l'espèce considérée et ses applications dans le cadre des investigations en cours sur la lutte génétique contre ce ravageur. Ce piège est cent fois plus attractif que les pièges traditionnels et a prouvé son efficacité dans les récentes études conduites sur le terrain.

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Second report of the OILB working group on genetic control ofR. cerasi.