Gergely Krett

Biofilm Bacterial Communities Inhabiting the Cave Walls of the Buda Thermal Karst System, Hungary

The diversity of biofilm bacterial communities associated with cave walls of the Buda Thermal Karst System (BTKS) located in Hungary was studied by scanning electron microscopy and molecular cloning based on 16S rRNA genes. Samples from two sites, the Molnár János cave (MJB) and the Rudas-Török spring cave (RTB), respectively, were analyzed and compared. The presence of iron precipitates was typical at both study sites, despite the fact that the cell morphological structure of the biofilms observed by SEM was characteristically different. Clones analyzed from BTKS were found to belong to 10 common phyla (Thermodesulfobacteria, Chloroflexi, Nitrospirae, Chlorobi, Proteobacteria, Firmicutes, Actinobacteria, Planctomycetes, Bacteroidetes, Verrucomicrobia) within the domain Bacteria. Moreover, sequences related to Aquificeae, Acidobacteria and Gemmatimonadetes were exclusive to MJB, while Cyanobacteria were found in RTB only. The phylogenetic distribution of the dominant bacterial clones was quite dissimilar between the two sites. In the biofilm from MJB clones affiliated with Firmicutes, whereas in the RTB clones related to Deltaproteobacteria were found in the highest number. In addition, substantially larger numbers of clone sequences related to thermophilic bacteria were recovered from MJB. On the basis of sequences of known microorganisms corresponding to our clone sequences, it is assumed that aerobic as well as anaerobic iron and sulfur transformation performed by different bacterial communities might be important biogenic processes in both caves.

Diversity and morphological structure of bacterial communities inhabiting the Diana-Hygieia Thermal Spring (Budapest, Hungary)

The Buda Thermal Karst System is an active hypogenic karst area that offers possibility for the analysis of biogenic cave formation. The aim of the present study was to gain information about morphological structure and genetic diversity of bacterial communities inhabiting the Diana-Hygieia Thermal Spring (DHTS). Using scanning electron microscopy, metal accumulating and unusual reticulated filaments were detected in large numbers in the DHTS biofilm samples. The phyla Actinobacteria, Firmicutes and Proteobacteria were represented by both bacterial strains and molecular clones but phyla Acidobacteria, Chlorobi, Chlorofexi, Gemmatimonadetes, Nitrospirae and Thermotogae only by molecular clones which showed the highest similarity to uncultured clone sequences originating from different environmental sources. The biofilm bacterial community proved to be somewhat more diverse than that of the water sample and the distribution of the dominant bacterial clones was different between biofilm and water samples. The majority of biofilm clones was affiliated with Deltaproteobacteria and Nitrospirae while the largest group of water clones was related to Betaproteobacteria. Considering the metabolic properties of known species related to the strains and molecular clones from DHTS, it can be assumed that these bacterial communities may participate in the local sulphur and iron cycles, and contribute to biogenic cave formation.

Winter Planktonic Microbial Communities in Highland Aquatic Habitats

ABSTRACT Winter conditions in aquatic habitats of the temperate zone markedly differ from those present in warmer seasons, nevertheless, relatively scarce information is available on planktonic microbial composition, as sites are not easily accessible and it was supposed traditionally that microbial activity is low during this cold period. Since microorganisms could have great impact on the ecosystem even during winter, we explored various sites in the Eastern Carpathians regarding the abundance and taxonomic composition of planktonic microorganisms. Although many of the studied environments were extreme habitats, planktonic microbial communities were abundant and mostly diverse with the presence of previously unidentified taxa.

A POLYPHASIC STUDY ON THE SPECIES DIVERSITY OF THE SEDIMENT MICROBIOTA OF LAKE HÉVÍZ

Lake Hévíz is the largest natural thermal lake of Europe, harboring special bacterial communities. The aim of the present study was to gain information about the distribution and species diversity of the sediment microbiota, with special focus on Actinobacteria, by using cultivation-based and -independent molecular methods. Samples from two depths were taken in two different locations in October 2007. 245 strains were isolated, grouped to 85 OTUs by ARDRA, and identified by 16S rDNA sequencing. Most of the strains showed highest sequence similarity with Bacillus and related genera. Strains belonging to the phylum Actinobacteria were identified as members of Arthrobacter, Brachybacterium, Brevibacterium, Curtobacterium, Friedmanniella, Gordonia, Kocuria, Microbacterium, Micrococcus, Micromonospora, Mycobacterium, Rhodococcus, Streptomyces and Williamsia . Two clone libraries were constructed from H3M and H4M samples, providing 288 and 192 clones which were grouped to 150 and 125 OTUs, respectively, by ARDRA. The two most abundant group of the H4M library were OP8-related. The phylum Proteobacteria was represented mostly by delta-Proteobacteria, other relevant groups were Cyanobacteria, Bacteroidetes, Acidobacteria and beta-Proteobacteria. The H3M library was dominated by Cyanobacteria, Verrucomicrobia, beta-Proteobacteria, gamma-Proteobacteria and delta-Proteobacteria. Chloroflexi, Bacteroidetes, Spirochetes and Firmicutes were scarce. Results from the clone libraries were compared to the length-heterogeneity-PCR fingerprints of the communities.

Gut content microbiota of introduced bigheaded carps (Hypophthalmichthys spp.) inhabiting the largest shallow lake in Central Europe

Studying the microbiota in the alimentary tract of bigheaded carps (Hypophthalmichthys spp.) gained special interest recently, as these types of investigations on non-native fish species may lead to a better understanding of their ecological role and feeding habits in an invaded habitat. For microbiological examinations, bigheaded carp gut contents and water column samples from Lake Balaton (Hungary) were collected from spring to autumn in 2013. Denaturing Gradient Gel Electrophoresis (DGGE) and pyrosequencing of the 16S rRNA gene were performed to reveal the composition. According to the DGGE patterns, bacterial communities of water samples separated clearly from that of the intestines. Moreover, the bacterial communities in the foreguts and hindguts were also strikingly dissimilar. Based on pyrosequencing, both foregut and hindgut samples were predominated by the fermentative genus Cetobacterium (Fusobacteria). The presence of some phytoplankton taxa and the high relative abundance of cellulose-degrading bacteria in the guts suggest that intestinal microbes may have an important role in digesting algae and making them utilizable for bigheaded carps that lack cellulase enzyme. In turn, the complete absence of typical heterotrophic freshwater bacteria in all studied sections of the intestines indicated that bacterioplankton probably has a negligible role in the nutrition of bigheaded carps.

Diversity and ecological tolerance of bacteria isolated from the rhizosphere of halophyton plants living nearby Kiskunság soda ponds, Hungary

Many halophytes and halophilic microorganisms are capable to adapt to the extremities of saline habitats. This study reveals the taxonomic diversity and ecological tolerance of bacteria isolated from the rhizosphere of three different halophytes (Bolboschoenus maritimus, Puccinellia limosa and Aster tripolium) living in the vicinity of Kiskunság soda ponds. Following a sampling in September 2013, altogether 76 bacterial strains were isolated using two different media. The strains were identified on the basis of 16S rRNA gene sequencing following ARDRA grouping. Salt and pH tolerance of the strains were examined by measuring their growth in broths containing 0-15% NaCl (w/V) and characterized with pH 7-12 values. Among the strains genera of Anaerobacillus, Bacillus and Exiguobacterium (Firmicutes), Agromyces, Isoptericola, Microbacterium, Micrococcus, Nocardiopsis, Nesterenkonia and Streptomyces (Actinobacteria), Halomonas and Idiomarina (Proteobacteria) and Anditalea (Bacteroidetes) were identified. The Bolboschoenus and Puccinellia samples characterized with the highest pH and electric conductivity values were dominated by Bacillus, Halomonas and Nesterenkonia, respectively. The salt tolerance of the bacterial strains was strongly dependent on the sampling location and plant species. In contrast, growth of bacterial strains in broths with alkaline pH values was more balanced. The strains from the Puccinellia sample showed the widest salt and pH tolerance.

Changes in the water quality and bacterial community composition of an alkaline and saline oxbow lake used for temporary reservoir of geothermal waters

Geothermal waters exploited in the southeastern region of Hungary are alkali-hydrogen-carbonate type, and beside the high amount of dissolved salt, they contain a variety of aromatic, heteroaromatic, and polyaromatic hydrocarbons. The majority of these geothermal waters used for heating are directed into surface waters following a temporary storage in reservoir lakes. The aim of this study was to gain information about the temporal and spatial changes of the water quality as well as the bacterial community composition of an alkaline and saline oxbow lake operated as reservoir of used geothermal water. On the basis of the water physical and chemical measurements as well as the denaturing gradient gel electrophoresis (DGGE) patterns of the bacterial communities, temporal changes were more pronounced than spatial differences. During the storage periods, the inflow, reservoir water, and sediment samples were characterized with different bacterial community structures in both studied years. The 16S ribosomal RNA (rRNA) gene sequences of the bacterial strains and molecular clones confirmed the differences among the studied habitats. Thermophilic bacteria were most abundant in the geothermal inflow, whereas the water of the reservoir was dominated by cyanobacteria and various anoxygenic phototrophic prokaryotes. In addition, members of several facultative anaerobic denitrifying, obligate anaerobic sulfate-reducing and syntrophic bacterial species capable of decomposition of different organic compounds including phenols were revealed from the water and sediment of the reservoir. Most of these alkaliphilic and/or halophilic species may participate in the local nitrogen and sulfur cycles and contribute to the bloom of phototrophs manifesting in a characteristic pink-reddish discoloration of the water of the reservoir.

Seasonal and spatial changes of planktonic bacterial communities inhabiting the natural thermal Lake Hévíz, Hungary

Lake Hévíz is a unique thermal spa located in Hungary. Owing to the thermal springs nourishing the lake, it has a relatively rapid water turnover. In spring 2011 a comprehensive embankment reconstruction was performed to preserve the water supply of the surrounding wetland habitats. The physical and chemical parameters as well as the planktonic microbial communities were studied with special respect to the effect of the disturbance of the water of Lake Hévíz. According to the abiotic components, both temporal and spatial differences were revealed with the exception of autumn samples. The reconstruction resulted in a short term but dramatic alteration of the total planktonic bacterial and cyanobacterial community structures as revealed by denaturing gradient gel electrophoresis. In addition, greater seasonal than spatial differences of bacterial communities were also observed. Planktonic bacterial community composition of Lake Hévíz included mainly typical freshwater species within phylum Actinobacteria, Chloroflexi, Cyanobacteria and class Alpha-, Beta- and Gamma-proteobacteria. Most of them were aerobic or facultative anaerobic heterotrophic but chemolitotrophic (e.g. Thiobacillus) or photolithotrophic (e.g. Cyanobacteria and Chloroflexi) autotrophic microbes were also identified.

Prokaryotic phylogenetic diversity of Hungarian deep subsurface geothermal well waters

Geothermal wells characterized by thermal waters warmer than 30°C can be found in more than 65% of the area of Hungary. The examined thermal wells located nearby Szarvas are used for heating industrial and agricultural facilities because of their relatively high hydrocarbon content. The aim of this study was to reveal the prokaryotic community structure of the water of SZR18, K87 and SZR21 geothermal wells using molecular cloning methods and Denaturing Gradient Gel Electrophoresis (DGGE). Water samples from the outflow pipes were collected in 2012 and 2013. The phylogenetic distribution of archaeal molecular clones was very similar in each sample, the most abundant groups belonged to the genera Methanosaeta, Methanothermobacter and Thermofilum. In contrast, the distribution of bacterial molecular clones was very diverse. Many of them showed the closest sequence similarities to uncultured clone sequences from similar thermal environments. From the water of the SZR18 well, phylotypes closely related to genera Fictibacillus and Alicyclobacillus (Firmicutes) were only revealed, while the bacterial diversity of the K87 well water was much higher. Here, the members of the phyla Thermodesulfobacteria, Proteobacteria, Nitrospira, Chlorobi, OP1 and OPB7 were also detected besides Firmicutes.

Phylogenetic diversity of bacterial communities inhabiting the sediment of Lake Hévíz — A comparison of cultivation and cloning

Lake Hévíz is the largest natural warm water lake of Europe. The curative mud of the lake comprises volcanic and marsh components although their species composition is hardly known yet. The aim of the present study was to gain information about the distribution and species diversity of bacterial communities inhabiting the sediment of Lake Hévíz using cultivation-based and molecular cloning methods. Samples from two depths and locations were taken in 2004 and 2007. Representatives of the altogether 255 bacterial isolates were affiliated with the phyla Firmicutes, Actinobacteria, Proteobacteria and Bacteroidetes. The most abundant groups belonged to the genus Bacillus (Firmicutes). Many of Lake Hévíz isolates showed the highest sequence similarity to bacteria known to be plant associated or members of normal human microbiota as well as participating in decomposition of highly resistant organic materials. In the three clone libraries, phylotypes belonging to altogether different phyla (Firmicutes, Actinobacteria, Proteobacteria, Bacteroidetes, Cyanobacteria, Chlorobi, Chloroflexi, Deferribacteres, Nitrospirae, Spirochaetes and Verrucomicrobia) were revealed from which members of Gammaproteobacteria and Cyanobacteria proved to be the most abundant. Regardless of the sampling times and methodology used, high spatial heterogeneities of bacterial community structures were characteristic of the sediment of Lake Hévíz.