Despite genetic isolation in sympatry, post-copulatory reproductive barriers have not evolved between bat- and human-associated common bedbugs (Cimex lectularius L.).

BACKGROUND: The common bedbug Cimex lectularius is a widespread ectoparasite on humans and bats. Two genetically isolated lineages, parasitizing either human (HL) or bat (BL) hosts, have been suggested to differentiate because of their distinct ecology. The distribution range of BL is within that of HL and bedbugs live mostly on synanthropic bat hosts. This sympatric co-occurrence predicts strong reproductive isolation at the post-copulatory level. RESULTS: We tested the post-copulatory barrier in three BL and three HL populations in reciprocal crosses, using a common-garden blood diet that was novel to both lineages. We excluded pre-copulation isolation mechanisms and studied egg-laying rates after a single mating until the depletion of sperm, and the fitness of the resulting offspring. We found a higher sperm storage capability in BL, likely reflecting the different seasonal availability of HL and BL hosts. We also observed a notable variation in sperm function at the population level within lineages and significant differences in fecundity and offspring fitness between lineages. However, no difference in egg numbers or offspring fitness was observed between within- and between-lineage crosses. CONCLUSIONS: Differences in sperm storage or egg-laying rates between HL and BL that we found did not affect reproductive isolation. Neither did the population-specific variation in sperm function. Overall, our results show no post-copulatory reproductive isolation between the lineages. How genetic differentiation in sympatry is maintained in the absence of a post-copulatory barrier between BL and HL remains to be investigated.

Complete genome sequencing and annotation of Rhodomicrobium vannielii strain DSM166 suggest affiliation to Rhodomicrobium lacus.

Rhodomicrobium vannielii is a multicellular and differentiating member of the order Hyphomicrobiales in the class Alphaproteobacteria. Here, we report the complete genome of strain DSM166 obtained by PacBio SMRT sequencing. The results suggest that this strain is closely related to Rhodomicrobium lacus.

Biodegradable polymers boost reproduction in the earthworm Eisenia fetida.

Microplastic contamination in soil has become a global environmental threat as it adversely affects terrestrial organisms like earthworms as well as soil properties. Especially biodegradable polymers have recently been used as an alternative to conventional polymer types, although their impact remains poorly understood. Thus, we studied the effect of conventional (polystyrene: PS, polyethylene terephthalate: PET, polypropylene: PP) versus aliphatic polyesters classified as biodegradable polymers (poly-(l-lactide): PLLA, polycaprolactone: PCL) on the earthworm Eisenia fetida and soil properties (pH and cation exchange capacity). We addressed direct effects on the weight gain and reproductive success of E. fetida, and indirect effects, like changes in the gut microbial composition as well as the production of short-chain fatty acids by the gut microbiota. Earthworms were exposed for eight weeks in an artificial soil amended with two environmentally relevant concentrations (1 % and 2.5 % (w/w)) of the different microplastic types. PLLA and PCL boosted the number of cocoons produced by 135 % and 54 %, respectively. Additionally, exposure to these two polymers increased number of hatched juveniles, changed gut microbial beta-diversity, and increased the production of the short chain fatty acid lactate compared to the control treatments. Interestingly, we also found a positive effect of PP on the earthworm's bodyweight and reproductive success. The interaction of microplastic and earthworms decreased soil pH by about 1.5 units in the presence of PLLA and PCL. No polymer effect on the cation exchange capacity of soil was found. In general, neither the presence of conventional nor biodegradable polymers had any adverse effects on any of the studied endpoints. Our results suggest that the effects of microplastic highly depend on the polymer type, and that the degradation of biodegradable polymers might be enhanced in the gut of earthworms, which implies that they may use biodegradable polymers as a potential carbon source.

Diesel exhaust particles alter gut microbiome and gene expression in the bumblebee Bombus terrestris.

Insect decline is a major threat to ecosystems around the world as they provide many important functions, such as pollination or pest control. Pollution is one of the main reasons for the decline, alongside changes in land use, global warming, and invasive species. While negative impacts of pesticides are well-studied, there is still a lack of knowledge about the effects of other anthropogenic pollutants, such as airborne particulate matter, on insects. To address this, we exposed workers of the bumblebee Bombus terrestris to sublethal doses of diesel exhaust particles (DEPs) and brake dust, orally or via air. After 7 days, we looked at the composition of the gut microbiome and tracked changes in gene expression. While there were no changes in the other treatments, oral DEP exposure significantly altered the structure of the gut microbiome. In particular, the core bacterium Snodgrassella had a decreased abundance in the DEP treatment. Similarly, transcriptome analysis revealed changes in gene expression after oral DEP exposure, but not in the other treatments. The changes are related to metabolism and signal transduction, which indicates a general stress response. Taken together, our results suggest potential health effects of DEP exposure on insects, here shown in bumblebees, as gut dysbiosis may increase the susceptibility of bumblebees to pathogens, while a general stress response may lower available energy resources. Those effects may exacerbate under natural conditions where insects face a multiple-stressor environment.

Microplastic ingestion affects hydrogen production and microbiomes in the gut of the terrestrial isopod Porcellio scaber.

Microplastic (MP) is an environmental burden and enters food webs via ingestion by macrofauna, including isopods (Porcellio scaber) in terrestrial ecosystems. Isopods represent ubiquitously abundant, ecologically important detritivores. However, MP-polymer specific effects on the host and its gut microbiota are unknown. We tested the hypothesis that biodegradable (polylactic acid [PLA]) and non-biodegradable (polyethylene terephthalate [PET]; polystyrene [PS]) MPs have contrasting effects on P. scaber mediated by changes of the gut microbiota. The isopod fitness after an 8-week MP-exposure was generally unaffected, although the isopods showed avoidance behaviour to PS-food. MP-polymer specific effects on gut microbes were detected, including a stimulation of microbial activity by PLA compared with MP-free controls. PLA stimulated hydrogen emission from isopod guts, while PET and PS were inhibitory. We roughly estimated 107 kg year-1 hydrogen emitted from the isopods globally and identified their guts as anoxic, significant mobile sources of reductant for soil microbes despite the absence of classical obligate anaerobes, likely due to Enterobacteriaceae-related fermentation activities that were stimulated by lactate generated during PLA-degradation. The findings suggest negative effects of PET and PS on gut fermentation, modulation of important isopod hydrogen emissions by MP pollution and the potential of MP to affect terrestrial food webs.

Optimization of soil microbial fuel cell for sustainable bio-electricity production: combined effects of electrode material, electrode spacing, and substrate feeding frequency on power generation and microbial community diversity.

BACKGROUND: Microbial fuel cells (MFCs) are among the leading research topics in the field of alternative energy sources due to their multifunctional potential. However, their low bio-energy production rate and unstable performance limit their application in the real world. Therefore, optimization is needed to deploy MFCs beyond laboratory-scale experiments. In this study, we investigated the combined influence of electrode material (EM), electrode spacing (ES), and substrate feeding interval (SFI) on microbial community diversity and the electrochemical behavior of a soil MFC (S-MFC) for sustainable bio-electricity generation. RESULTS: Two EMs (carbon felt (CF) and stainless steel/epoxy/carbon black composite (SEC)) were tested in an S-MFC under three levels of ES (2, 4, and 8 cm) and SFI (4, 6, and 8 days). After 30 days of operation, all MFCs achieved open-circuit voltage in the range of 782 + 12.2 mV regardless of the treatment. However, the maximum power of the SEC-MFC was 3.6 times higher than that of the CF-MFC under the same experimental conditions. The best solution, based on the interactive influence of the two discrete variables, was obtained with SEC at an ES of 4.31 cm and an SFI of 7.4 days during an operating period of 66 days. Analysis of the experimental treatment effects of the variables revealed the order SFI < ES < EM, indicating that EM is the most influential factor affecting the performance of S-MFC. The performance of S-MFC at a given ES value was found to be dependent on the levels of SFI with the SEC electrode, but this interactive influence was found to be insignificant with the CF electrode. The microbial bioinformatic analysis of the samples from the S-MFCs revealed that both electrodes (SEC and CF) supported the robust metabolism of electroactive microbes with similar morphological and compositional characteristics, independent of ES and SFI. The complex microbial community showed significant compositional changes at the anode and cathode over time. CONCLUSION: This study has demonstrated that the performance of S-MFC depends mainly on the electrode materials and not on the diversity of the constituent microbial communities. The performance of S-MFCs can be improved using electrode materials with pseudocapacitive properties and a larger surface area, instead of using unmodified CF electrodes commonly used in S-MFC systems.

Microplastic polymer properties as deterministic factors driving terrestrial plastisphere microbiome assembly and succession in the field.

Environmental microplastic (MP) is ubiquitous in aquatic and terrestrial ecosystems providing artificial habitats for microbes. Mechanisms of MP colonization, MP polymer impacts, and effects on soil microbiomes are largely unknown in terrestrial systems. Therefore, we experimentally tested the hypothesis that MP polymer type is an important deterministic factor affecting MP community assembly by incubating common MP polymer types in situ in landfill soil for 14 months. 16S rRNA gene amplicon sequencing indicated that MP polymers have specific impacts on plastisphere microbiomes, which are subsets of the soil microbiome. Chloroflexota, Gammaproteobacteria, certain Nitrososphaerota, and Nanoarchaeota explained differences among MP polymers and time points. Plastisphere microbial community composition derived from different MP diverged over time and was enriched in potential pathogens. PICRUSt predictions of pathway abundances and quantitative PCR of functional marker genes indicated that MP polymers exerted an ambivalent effect on genetic potentials of biogeochemical cycles. Overall, the data indicate that (i) polymer type as deterministic factor rather than stochastic factors drives plastisphere community assembly, (ii) MP impacts greenhouse gas metabolism, xenobiotic degradation and pathogen distribution, and (iii) MP serves as an ideal model system for studying fundamental questions in microbial ecology such as community assembly mechanisms in terrestrial environments.

Influence of NH3 and NH4+ on anaerobic digestion and microbial population structure at increasing total ammonia nitrogen concentrations.

Despite the extensive research dedicated to ammonia inhibition, the effect of NH3 and NH4+ on each anaerobic digestion stage and the associated microorganisms is still not completely understood. In the past, the focus was mainly on methanogenesis and either on NH3 or total ammonia nitrogen (TAN). Here, anaerobic digestion of two defined substrates, namely starch/NH4Cl and casein, was investigated particularly regarding the effects of different NH3/NH4+ ratios on the involved microorganisms. TAN affected bacteria, primarily gram-positive ones, whereas archaea responded largely to the NH3 concentration. These sensitivity differences are attributed to differences in the corresponding cell-membrane structures. A TAN decrease via stripping performed in two full-scale agricultural biogas plants resulted in increased bacterial diversity, with a pronounced increase in the propionate acetogens' abundance. Based on these data, it is suggested that inhibition can be avoided and processes stabilized in biogas plants by adjusting the NH3/NH4+ ratio, when feeding nitrogen-rich substrates.

Pristine and artificially-aged polystyrene microplastic particles differ in regard to cellular response.

Microplastic particles (MP), arising from the gradual decomposition of plastics in the environment, have been identified as a global problem. Most investigations of MP cytotoxicity use pristine spherical particles available from commercial sources when evaluating their impact on mammalian cells, while only limited data is available for the more relevant "weathered microplastic". In this study, we exposed murine macrophages to polystyrene MP either after up to 130 days of accelerated ageing or in pristine condition. Weathered and pristine MP were physicochemically characterized, and their cytotoxicity was investigated using biological assays, transcriptome analysis, and metabolic pathways prediction. Whereas the response to pristine MP is mainly dominated by a TNF-α release, sharp-edged weathered MP induce broader adverse cellular reactions. This study stresses the importance of including more realistic test particles (e.g., weathered particles) in combination with a broad range of biological assays when evaluating the potential risk of microplastic exposure.

In situ Prokaryotic and Eukaryotic Communities on Microplastic Particles in a Small Headwater Stream in Germany.

The ubiquitous use of plastic products in our daily life is often accompanied by improper disposal. The first interactions of plastics with organisms in the environment occur by overgrowth or biofilm formation on the particle surface, which can facilitate the ingestion by animals. In order to elucidate the colonization of plastic particles by prokaryotic and eukaryotic microorganisms in situ, we investigated microbial communities in biofilms on four different polymer types and on mineral particles in a small headwater stream 500 m downstream of a wastewater treatment plant in Germany. Microplastic and mineral particles were exposed to the free-flowing water for 4 weeks in spring and in summer. The microbial composition of the developing biofilm was analyzed by 16S and 18S amplicon sequencing. Despite the expected seasonal differences in the microbial composition of pro- and eukaryotic communities, we repeatedly observed polymer type-specific differentiation in both seasons. The order of polymer type-specific prokaryotic and eukaryotic community distances calculated by Robust Aitchison principal component analysis (PCA) was the same in spring and summer samples. However, the magnitude of the distance differed considerably between polymer types. Prokaryotic communities on polyethylene particles exhibited the most considerable difference to other particles in summer, while eukaryotic communities on polypropylene particles showed the most considerable difference to other spring samples. The most contributing bacterial taxa to the polyethylene-specific differentiation belong to the Planctomycetales, Saccharimonadales, Bryobacterales, uncultured Acidiomicrobia, and Gemmatimonadales. The most remarkable differences in eukaryotic microorganism abundances could be observed in several distinct groups of Ciliophora (ciliates) and Chlorophytes (green algae). Prediction of community functions from taxonomic abundances revealed differences between spring and summer, and - to a lesser extent - also between polymer types and mineral surfaces. Our results show that different microplastic particles were colonized by different biofilm communities. These findings may be used for advanced experimental designs to investigate the role of microorganisms on the fate of microplastic particles in freshwater ecosystems.

Male diet affects female fitness and sperm competition in human- and bat-associated lineages of the common bedbug, Cimex lectularius.

Sperm performance can vary in ecologically divergent populations, but it is often not clear whether the environment per se or genomic differences arising from divergent selection cause the difference. One powerful and easily manipulated environmental effect is diet. Populations of bedbugs (Cimex lectularius) naturally feed either on bat or human blood. These are diverging genetically into a bat-associated and a human-associated lineage. To measure how male diet affects sperm performance, we kept males of two HL and BL populations each on either their own or the foreign diet. Then we investigated male reproductive success in a single mating and sperm competition context. We found that male diet affected female fecundity and changed the outcome of sperm competition, at least in the human lineage. However, this influence of diet on sperm performance was moulded by an interaction. Bat blood generally had a beneficial effect on sperm competitiveness and seemed to be a better food source in both lineages. Few studies have examined the effects of male diet on sperm performance generally, and sperm competition specifically. Our results reinforce the importance to consider the environment in which sperm are produced. In the absence of gene flow, such differences may increase reproductive isolation. In the presence of gene flow, however, the generally better sperm performance after consuming bat blood suggests that the diet is likely to homogenise rather than isolate populations.

Keeping the shoot above water - submergence triggers antithetical growth responses in stems and petioles of watercress (Nasturtium officinale).

The molecular mechanisms controlling underwater elongation are based extensively on studies on internode elongation in the monocot rice (Oryza sativa) and petiole elongation in Rumex rosette species. Here, we characterize underwater growth in the dicot Nasturtium officinale (watercress), a wild species of the Brassicaceae family, in which submergence enhances stem elongation and suppresses petiole growth. We used a genome-wide transcriptome analysis to identify the molecular mechanisms underlying the observed antithetical growth responses. Though submergence caused a substantial reconfiguration of the petiole and stem transcriptome, only little qualitative differences were observed between both tissues. A core submergence response included hormonal regulation and metabolic readjustment for energy conservation, whereas tissue-specific responses were associated with defense, photosynthesis, and cell wall polysaccharides. Transcriptomic and physiological characterization suggested that the established ethylene, abscisic acid (ABA), and GA growth regulatory module for underwater elongation could not fully explain underwater growth in watercress. Petiole growth suppression is likely attributed to a cell cycle arrest. Underwater stem elongation is driven by an early decline in ABA and is not primarily mediated by ethylene or GA. An enhanced stem elongation observed in the night period was not linked to hypoxia and suggests an involvement of circadian regulation.

Flexible feeding in anaerobic digestion - Impact on process stability, performance and microbial community structures.

Biogas has the potential to contribute to some of the most urgent issues of the energy transition, including mobility, energy storage, and grid stability. Flexibilization has been discussed as a means to improve the economics of biogas production, ideally restricting the production of electricity to times of strong need. Here the possibility of demand-driven, flexible biogas production is investigated, which saves substrates and storage capacity, while still enabling control over the production of electricity. Effects of different flexible feeding regimes were tested in a continuously operated 200 L reactor. After a period of 300 days under steady conditions (6.4 kg feed m-3d-1), varying flexible feeding patterns were applied over the next 700 days. Biogas production, volatile organic acid concentrations, and microbial dynamics were documented. Reduction of feeding resulted in reducing the gas production by up to 80% within a day. By increasing the feed, gas production could rapidly be reinitiated at similar levels as before even after fasting periods of up to 22 days. CH4-contents of the produced biogas were nearly constant over the investigation period. As a response to the flexible feeding, a reorganization of the microbial community was observed, which came to an end after 800 days and then was no longer affected by further changes in the feeding patterns or the substrate composition. Dominating archaea were of the order Methanosarcinales. During the experiment, representatives from the class Methanosaetaceae replaced representatives from the class Methanosarcinaceae.

Genome-Wide Identification of Essential and Auxiliary Gene Sets for Magnetosome Biosynthesis in Magnetospirillum gryphiswaldense.

Magnetotactic bacteria (MTB) stand out by their ability to manufacture membrane-enclosed magnetic organelles, so-called magnetosomes. Previously, it has been assumed that a genomic region of approximately 100 kbp, the magnetosome island (MAI), harbors all genetic determinants required for this intricate biosynthesis process. Recent evidence, however, argues for the involvement of additional auxiliary genes that have not been identified yet. In the present study, we set out to delineate the full gene complement required for magnetosome production in the alphaproteobacterium Magnetospirillum gryphiswaldense using a systematic genome-wide transposon mutagenesis approach. By an optimized procedure, a Tn5 insertion library of 80,000 clones was generated and screened, yielding close to 200 insertants with mild to severe impairment of magnetosome biosynthesis. Approximately 50% of all Tn5 insertion sites mapped within the MAI, mostly leading to a nonmagnetic phenotype. In contrast, in the majority of weakly magnetic Tn5 insertion mutants, genes outside the MAI were affected, which typically caused lower numbers of magnetite crystals with partly aberrant morphology, occasionally combined with deviant intracellular localization. While some of the Tn5-struck genes outside the MAI belong to pathways that have been linked to magnetosome formation before (e.g., aerobic and anaerobic respiration), the majority of affected genes are involved in so far unsuspected cellular processes, such as sulfate assimilation, oxidative protein folding, and cytochrome c maturation, or are altogether of unknown function. We also found that signal transduction and redox functions are enriched in the set of Tn5 hits outside the MAI, suggesting that such processes are particularly important in support of magnetosome biosynthesis.IMPORTANCE Magnetospirillum gryphiswaldense is one of the few tractable model magnetotactic bacteria (MTB) for studying magnetosome biomineralization. So far, knowledge on the genetic determinants of this complex process has been mainly gathered using reverse genetics and candidate approaches. In contrast, nontargeted forward genetics studies are lacking, since application of such techniques in MTB has been complicated for a number of technical reasons. Here, we report on the first comprehensive transposon mutagenesis study in MTB, aiming at systematic identification of auxiliary genes necessary to support magnetosome formation in addition to key genes harbored in the magnetosome island (MAI). Our work considerably extends the candidate set of novel subsidiary determinants and shows that the full gene complement underlying magnetosome biosynthesis is larger than assumed. In particular, we were able to define certain cellular pathways as specifically important for magnetosome formation that have not been implicated in this process so far.

Assessment of prokaryote to eukaryote ratios in environmental samples by SSU rDNA length polymorphism.

Microbial communities are important regulators of many processes in all ecosystems. Understanding of ecosystem processes requires at least an overview of the involved microorganisms. While in-depth identification of microbial species in environmental samples can be achieved by next generation sequencing, profiling of whole microbial communities can be accomplished via less labour-intensive approaches. Especially automated ribosomal intergenic spacer analysis (ARISA) are of interest as they are highly specific even at fine scales and widely applicable for environmental samples. Yet, established protocols lack the possibility to compare prokaryotic and eukaryotic communities as different primer sets are necessary. However, shifts in the eukaryote to prokaryote ratio can be a useful indicator for ecosystem processes like decomposition or nutrient cycling. We propose a protocol to analyse prokaryotic and eukaryotic communities using a single primer pair based reaction based on a region with variable length (V4, which is about 180 bp shorter in prokaryotes compared to eukaryotes) in the small ribosomal subunit flanked by two highly conservative regions. Shifts in the prokaryotic and eukaryotic ratio between samples can be reliably detected by fragment length polymorphism analysis as well as sequencing of this region. Together with established approaches such as ARISA or 16S and ITS rDNA sequencing, this can provide a more complex insight into microbial community shifts and ecosystem processes.

Arsenic metabolism in technical biogas plants: possible consequences for resident microbiota and downstream units.

The metal(loid) and in particular the Arsenic (As) burden of thirteen agricultural biogas plants and two sewage sludge digesters were investigated together with the corresponding microbial consortia. The latter were characterized by ARISA (automated ribosomal intergenetic spacer analysis) and next generation sequencing. The consortia were found to cluster according to digester type rather than substrate or metal(loid) composition. For selected plants, individual As species in the liquid and gaseous phases were quantified, showing that the microorganisms actively metabolize and thereby remove the As from their environment via the formation of (methylated) volatile species. The As metabolites showed some dependency on the microbial consortia, while there was no statistical correlation with the substrate mix. Finally, slurry from one agricultural biogas plant and one sewage sludge digester was transferred into laboratory scale reactors ("satellite reactors") and the response to a defined addition of As (30 and 60 µM sodium arsenite) was studied. The results corroborate the hypothesis of a rapid conversion of dissolved As species into volatile ones. Methanogenesis was reduced during that time, while there was no discernable toxic effect on the microbial population. However, the utilization of the produced biogas as replacement for natural gas, e.g. as fuel, may be problematic, as catalysts and machinery are known to suffer from prolonged exposure even to low As concentrations.

AtERF#111/ABR1 is a transcriptional activator involved in the wounding response.

AtERF#111/ABR1 belongs to the group X of the ERF/AP2 transcription factor family (GXERFs) and is shoot specifically induced under submergence and hypoxia. It was described to be an ABA-response repressor, but our data reveal a completely different function. Surprisingly, AtERF#111 expression is strongly responsive to wounding stress. Expression profiling of ERF#111-overexpressing (OE) plants, which show morphological phenotypes like increased root hair length and number, strengthens the hypothesis of AtERF#111 being involved in the wounding response, thereby acting as a transcriptional activator of gene expression. Consistent with a potential function outside of oxygen signalling, we could not assign AtERF#111 as a target of the PRT6 N-degron pathway, even though it starts with a highly conserved N-terminal Met-Cys (MC) motif. However, the protein is unstable as it is degraded in an ubiquitin-dependent manner. Finally, direct target genes of AtERF#111 were identified by microarray analyses and subsequently confirmed by protoplast transactivation assays. The special roles of diverse members of the plant-specific GXERFs in coordinating stress signalling and wound repair mechanisms have been recently hypothesized, and our data suggest that AtERF#111 is indeed involved in these processes.

Vitamin B2 (riboflavin) increases drought tolerance of Agaricus bisporus.

Drought is a stressor for many soil-inhabiting organisms. Although plants have been extensively investigated for drought-adaptive mechanisms, little information is available for fungi. Antioxidants are especially relevant, since desiccation is accompanied by an excessive intracellular production of reactive oxygen species. Riboflavin (vitamin B2) is one antioxidant regulating drought tolerance in plants. A similar function may exist in fungi. Here, we examined the respiratory and transcriptional responses of Agaricus bisporus to drought and the impact of riboflavin. Mesocosm experiments with four groups were established: hyphae were treated with or without 50 µM riboflavin under drought or no drought conditions. Drought increased riboflavin content in hyphae about 5 times with, but also without, addition of riboflavin. Without addition of riboflavin, fungal respiration decreased by more than 50% at water potentials of about -20 MPa. With addition of riboflavin, respiration remained about 2-3 times higher. The transcriptional responses to only drought or only riboflavin strongly overlapped and were mainly based on factors regulating transcription and translation. This was even stronger in combined treatments. Riboflavin induced protective mechanisms in drought-stressed hyphae. Most pronounced was the methylglyoxal (cytotoxic by-product of glycolysis) detoxifying of lactoylglutathione lyase. Thus, our data suggest a stress-priming function and a role of riboflavin in drought responses of A. bisporus.

Process parameters and changes in the microbial community patterns during the first 240 days of an agricultural energy crop digester.

Commercial biogas production takes place by complex microbial communities enclosed in controlled "technical ecosystems". Once established, the communities tend to be resilient towards disturbances, although the relative abundance of their members may vary. The start-up phase, during which the community establishes itself, is therefore decisive for the later performance of the reactor. In this study, we followed the first 240 days of a standard agricultural energy crop digester consisting of a 400 m(3) plug flow fermenter and a 1000 m(3) agitated post digester, operated at 40-45 °C. The feed consisted of corn and later grass silage augmented by ground wheat. Changes in both the eubacterial and methanogenic archaeal communities were followed by automated ribosomal intergenic spacer analysis (ARISA). In addition the copy number of the methyl-coenzyme reductase A (mcrA)-genes found in all known methanogens were followed by quantitative PCR, while selected samples from two phases-one early, one late-of the community structure development were subjected to high throughput sequencing. Biogas volume and composition (CH4, CO2, H2, H2S, O2), pH, ammonia-N, and volatile fatty acids (VFA), were measured as part of the routine process control. VFA/TIC values were calculated on this basis. Whereas the total gas production of the plant established itself at about 2500 m(3) biogas per day within the first months, the composition of the microbial communities showed distinct spatial and temporal differences over the investigated time period. Absolute values for DNA isolation procedures are difficult to certify, hence comparative results on community structures obtained using standardized ARISA with identical primers are of value. Moreover, ARISA patterns can be statistically analyzed to identify distinct subgroups and transitions between them as well as serial correlations. Thereby the microbial community and its structural development can be correlated with statistical relevance to changes in operational (feed) and process parameters (pH-value, biogas composition). In particular when augmented by deep sequencing data of judiciously chosen samples, this allows a hitherto unknown level of insight into the performance of technical biogas plants.

Spatial and ecological population genetic structures within two island-endemic Aeonium species of different niche width.

The Crassulacean genus Aeonium is a well-known example for plant species radiation on oceanic archipelagos. However, while allopatric speciation among islands is documented for this genus, the role of intra-island speciation due to population divergence by topographical isolation or ecological heterogeneity has not yet been addressed. The aim of this study was to investigate intraspecific genetic structures and to identify spatial and ecological drivers of genetic population differentiation on the island scale. We analyzed inter simple sequence repeat variation within two island-endemic Aeonium species of La Palma: one widespread generalist that covers a large variety of different habitat types (Ae. davidbramwellii) and one narrow ecological specialist (Ae. nobile), in order to assess evolutionary potentials on this island. Gene pool differentiation and genetic diversity patterns were associated with major landscape structures in both species, with phylogeographic implications. However, overall levels of genetic differentiation were low. For the generalist species, outlier loci detection and loci-environment correlation approaches indicated moderate signatures of divergent selection pressures linked to temperature and precipitation variables, while the specialist species missed such patterns. Our data point to incipient differentiation among populations, emphasizing that ecological heterogeneity and topographical structuring within the small scales of an island can foster evolutionary processes. Very likely, such processes have contributed to the radiation of Aeonium on the Canary Islands. There is also support for different evolutionary mechanisms between generalist and specialist species.