Kinetic characterization of a new phenol degrading Acinetobacter towneri strain isolated from landfill leachate treating bioreactor.

The objective of this study was to establish and to mathematically describe the phenol degrading properties of a new Acinetobacter towneri CFII-87 strain, isolated from a bioreactor treating landfill leachate. For this purpose, the biokinetic parameters of phenol biodegradation at various initial phenol concentrations of the A. towneri CFII-87 strain have been experimentally measured, and four different mathematical inhibition models (Haldane, Yano, Aiba and Edwards models) have been used to simulate the substrate-inhibited phenol degradation process. The results of the batch biodegradation experiments show that the new A. towneri CFII-87 strain grows on and metabolizes phenol up to 1000 mg/L concentration, manifests significant substrate inhibition and lag time only at concentrations above 800 mg/L phenol, and has a maximum growth rate at 300 mg/L initial phenol concentration. The comparison of the model predictions with the experimental phenol and biomass data revealed that the Haldane, Aiba and Edwards models can be used with success to describe the phenol biodegradation process by A. towneri CFII-87, while the Yano model, especially at higher initial phenol concentrations, fails to describe the process. The best performing inhibition model was the Edwards model, presenting correlation coefficients of R2 > 0.98 and modelling efficiency of ME > 0.94 for the prediction of biomass and phenol concentrations on the validation datasets. The calculated biokinetic model parameters place this new strain among the bacteria with the highest tolerance towards phenol. The results suggest that the A. towneri CFII-87 strain can potentially be used in the treatment of phenolic wastewaters.

High Coverage Mitogenomes and Y-Chromosomal Typing Reveal Ancient Lineages in the Modern-Day Székely Population in Romania.

Here we present 115 whole mitogenomes and 92 Y-chromosomal Short Tandem Repeat (STR) and Single Nucleotide Polymorphism (SNP) profiles from a Hungarian ethnic group, the Székelys (in Romanian: Secuii, in German: Sekler), living in southeast Transylvania (Romania). The Székelys can be traced back to the 12th century in the region, and numerous scientific theories exist as to their origin. We carefully selected sample providers that had local ancestors inhabiting small villages in the area of Odorheiu Secuiesc/Székelyudvarhely in Romania. The results of our research and the reported data signify a qualitative leap compared to previous studies since it presents the first complete mitochondrial DNA sequences and Y-chromosomal profiles of 23 STRs from the region. We evaluated the results with population genetic and phylogenetic methods in the context of the modern and ancient populations that are either geographically or historically related to the Székelys. Our results demonstrate a predominantly local uniparental make-up of the population that also indicates limited admixture with neighboring populations. Phylogenetic analyses confirmed the presumed eastern origin of certain maternal (A, C, D) and paternal (Q, R1a) lineages, and, in some cases, they could also be linked to ancient DNA data from the Migration Period (5th-9th centuries AD) and Hungarian Conquest Period (10th century AD) populations.

Endosymbiotic Bacterial Diversity of Corn Leaf Aphid, Rhopalosiphum maidis Fitch (Hemiptera: Aphididae) Associated with Maize Management Systems.

In this study, different maize fields cultivated under different management systems were sampled to test corn leaf aphid, Rhopalosiphum maidis, populations in terms of total and endosymbiotic bacterial diversity. Corn leaf aphid natural populations were collected from traditionally managed maize fields grown under high agricultural and natural landscape diversity as well as conventionally treated high-input agricultural fields grown in monoculture and with fertilizers use, hence with low natural landscape diversity. Total bacterial community assessment by DNA sequencing was performed using the Illumina MiSeq platform. In total, 365 bacterial genera were identified and 6 endosymbiont taxa. A high abundance of the primary endosymbiont Buchnera and secondary symbionts Serratia and Wolbachia were detected in all maize crops. Their frequency was found to be correlated with the maize management system used, probably with fertilizer input. Three other facultative endosymbionts ("Candidatus Hamiltonella", an uncultured Rickettsiales genus, and Spiroplasma) were also recorded at different frequencies under the two management regimes. Principal components analyses revealed that the relative contribution of the obligate and dominant symbiont Buchnera to the aphid endosymbiotic bacterial community was 72%, whereas for the managed system this was only 16.3%. When facultative symbionts alone were considered, the effect of management system revealed a DNA diversity of 23.3%.

Anion-type modulates the effect of salt stress on saline lake bacteria.

Beside sodium chloride, inland saline aquatic systems often contain other anions than chloride such as hydrogen carbonate and sulfate. Our understanding of the biological effects of salt composition diversity is limited; therefore, the aim of this study was to examine the effect of different anions on the growth of halophilic bacteria. Accordingly, the salt composition and concentration preference of 172 strains isolated from saline and soda lakes that differed in ionic composition was tested using media containing either carbonate, chloride or sulfate as anion in concentration values ranging from 0 to 0.40 mol/L. Differences in salt-type preference among bacterial strains were observed in relationship to the salt composition of the natural habitat they were isolated from indicating specific salt-type adaptation. Sodium carbonate represented the strongest selective force, while majority of strains was well-adapted to growth even at high concentrations of sodium sulfate. Salt preference was to some extent associated with taxonomy, although variations even within the same bacterial species were also identified. Our results suggest that the extent of the effect of dissolved salts in saline lakes is not limited to their concentration but the type of anion also substantially impacts the growth and survival of individual microorganisms.

In Vitro and In Vivo Effect of Poplar Bud Extracts on Phytophthora infestans: A New Effective Biological Method in Potato Late Blight Control.

The effect of populin extract from black poplar (Populus nigra) on seven different late blight strains was tested under laboratory and field conditions. The growth rate of hyphae was found to be significantly lower in vitro after 3 and 4 v/v% populin applications. Stain M16 was resistant to populin treatment under lab conditions, however. Both 5% and 10% concentration populin reduced the M16 strain's severity on potato leaves under field conditions and proved to be even more effective than conventionally used fungicides Infinito 687 Sc and Valis M. Higher infection intensity at the 1% level was observed after 24 h using Valis M, and the same trend toward 10% infection remained after 48 and 72 h as well. Low, almost-no-infection intensity was detected after populin 5% and 10% treatment under an open field condition. Altogether, it can be concluded that populin extract can be a low-cost option for growers and an environmentally friendly approach in late blight control.

Sapientia aquatica gen. nov., sp. nov., isolated from a crater lake.

A new aerobic betaproteobacterium, strain SA-152T, was isolated from the water of a crater lake. 16S rRNA gene sequence analysis revealed that strain SA-152T belonged to the family Oxalobacteraceae (order Burkholderiales) and was phylogenetically related to Solimicrobium silvestre S20-91T with 97.09 % and to Herminiimonas arsenicoxydans ULPAs1T with 96.00 % 16S rRNA gene pairwise sequence similarity. Cells of strain SA-152T were rod-shaped, non-motile, oxidase-negative and catalase-positive. Its fatty acid profile was dominated by two fatty acids, C16 : 1 ω7c and C16 : 0, the major respiratory quinones were Q-8 and Q-7, and the main polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The G+C content of the genomic DNA of strain SA-152T was 48.3 mol%. The new bacterium can be distinguished from closely related genera Solimicrobium, Herminiimonas, Rugamonas and Undibacterium based on its non-motile and oxidase-negative cells. On the basis of the phenotypic, chemotaxonomic and genomic data, strain SA-152T is considered to represent a novel species of a new genus, for which the name Sapientia aquatica gen. nov., sp. nov. is proposed. The type strain of Sapientia aquatica is SA-152T (=DSM 29805T=NCAIM B.02613T).

Siculibacillus lacustris gen. nov., sp. nov., a new rosette-forming bacterium isolated from a freshwater crater lake (Lake St. Ana, Romania).

A new aerobic alphaproteobacterium, strain SA-279T, was isolated from a water sample of a crater lake. The 16S rRNA gene sequence analysis revealed that strain SA-279T formed a distinct lineage within the family Ancalomicrobiaceae and shared the highest pairwise similarity values with Pinisolibacterravus E9T (96.4 %) and Ancalomicrobiumadetum NBRC 102456T (94.2 %). Cells of strain SA-279T were rod-shaped, motile, oxidase and catalase positive, and capable of forming rosettes. Its predominant fatty acids were C18 : 1ω7c (69.0 %) and C16 : 1ω7c (22.7 %), the major respiratory quinone was Q-10, and the main polar lipids were phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine, phosphatidylglycerol, an unidentified aminophospholipid and an unidentified lipid. The G+C content of the genomic DNA of strain SA-279T was 69.2 mol%. On the basis of the phenotypic, chemotaxonomic and molecular data, strain SA-279T is considered to represent a new genus and species within the family Ancalomicrobiaceae, for which the name Siculibacillus lacustris gen. nov., sp. nov. is proposed. The type strain is SA-279T (=DSM 29840T=JCM 31761T).

A new Rhizobium species isolated from the water of a crater lake, description of Rhizobium aquaticum sp. nov.

A novel isolate, strain SA-276T, was isolated from the water of Lake St. Ana, a crater lake which is located in Romania. Phylogenetic analysis based on the 16S rRNA gene revealed that the new strain is a member of the family Rhizobiaceae, showing a high pairwise similarity value (97.65%) to Rhizobium tubonense CCBAU 85046T (= DSM 25379T), Rhizobium leguminosarum USDA 2370T (= LMG 14904T), Rhizobium anhuiense CCBAU 23252T and Rhizobium laguerreae FB206T. Cells of strain SA-276T were rod-shaped, motile, oxidase negative and weakly catalase positive. The predominant fatty acids were C18:1ω7c and cyclo C19:0ω8c, the major respiratory quinones were Q-10 and Q-9, and the main polar lipids were phosphatidylmonomethylethanolamine, phosphatidylglycerol and phosphatidylcholine. The G + C content of the genomic DNA of strain SA-276T was 60.8 mol%. The novel isolate can be distinguished from the closest related type strain R. tubonense DSM 25379T based on its broader substrate specificity and positive trypsin enzyme activity. On the basis of the phenotypic, chemotaxonomic and molecular data, strain SA-276T is considered to represent a new species, for which the name Rhizobium aquaticum sp. nov. is proposed. The type strain is SA-276T (= DSM 29780T = JCM 31760T).

Quisquiliibacterium transsilvanicum gen. nov., sp. nov., a novel betaproteobacterium isolated from a waste-treating bioreactor.

A new betaproteobacterium, CGI-09T, was isolated from an activated sludge bioreactor which treated landfill leachate. Based on 16S rRNA gene sequence analysis, the new strain shared the highest pairwise similarity values with members of the order Burkholderiales: Derxia gummosa IAM 13946T (family Alcaligenaceae), 93.7 % and Lautropia mirabilis DSM 11362T (family Burkholderiaceae), 93.6 %. Cells of strain CGI-09T were rod-shaped and non-motile. The new strain was oxidase and catalase positive and capable of reducing nitrate to nitrite. The predominant fatty acids were C16 : 1 ω7c, C16 : 0, cycloC17 : 0 and C18 : 1 ω7c, the major respiratory quinone was Q-8, and the detected polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unknown phospholipid. The G+C content of the genomic DNA of strain CGI-09T was 70.2 mol%. The new bacterium can be distinguished from the members of genera Derxia and Lautropia based on its non-motile cells, arginine dihydrolase activity, its high cyclo C17 : 0 fatty acid content and the lack of hydroxy fatty acids. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CGI-09T is considered to represent a new genus and species within the family Burkholderiaceae, for which the name Quisquiliibacterium transsilvanicum gen. nov., sp. nov. is proposed. The type strain is CGI-09T (=DSM 29781T=JCM 31785T).

Caenimicrobium hargitense gen. nov., sp. nov., a new member of the family Alcaligenaceae (Betaproteobacteria) isolated from activated sludge.

A new betaproteobacterium, CGII-59m2T, was isolated from an activated sludge bioreactor which treated landfill leachate. The 16S rRNA gene sequence analysis revealed that strain CGII-59m2T belonged to the family Alcaligenaceae and shared the highest pairwise similarity values with Parapusillimonas granuli LMG 24012T (97.7 %), various species of the genus Bordetella (97.3-97.0 %) and Candidimonas nitroreducens LMG 24812T (97.0 %). Cells of strain CGII-59m2T were rod-shaped, non-motile, and oxidase- and catalase-positive. The predominant fatty acids were C16 : 1ω7c, C16 : 0, cyclo C17 : 0 and C18 : 1ω7c, the major respiratory quinone was Q-8, and the main polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown phospholipid. The G+C content of the genomic DNA of strain CGII-59m2T was 62.3 mol%. The new bacterium can be distinguished from the closely related type strains based on its non-motile cells and its high C16 : 1ω7c fatty acid content. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CGII-59m2T is considered to represent a novel species of a new genus, for which the name Caenimicrobium hargitense gen. nov., sp. nov. is proposed. The type strain is CGII-59m2T (=DSM 29806T=NCAIM B.02615T).

Rhodococcus sovatensis sp. nov., an actinomycete isolated from the hypersaline and heliothermal Lake Ursu.

A Gram-stain-positive, strictly aerobic, mesophilic bacterium, designated H004T, was isolated from a water sample of the hypersaline and heliothermal Lake Ursu, Sovata, Romania. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain H004T formed a distinct phyletic lineage within the genus Rhodococcus. It shared the highest 16S rRNA gene sequence similarity with Rhodococcus yunnanensis YIM 70056T (98.80 %), followed by Rhodococcus fascians LMG 3623T (98.73 %), Rhodococcus cercidiphylli YIM 65003T (98.73 %), Rhodococcus cerastii C5T (98.58 %) and Rhodococcus kyotonensis DS472T (98.53 %). The alkB-based phylogenetic analysis further confirmed that this strain constitutes a highly unique lineage within the genus. Chemotaxonomic characteristics, including the predominant fatty acids acids C15 : 0, C18 : 1ω9c, C19 : 1ω11c/C19 : 1ω9c and C16 : 1ω7c/iso-C15 : 0 2-OH, the major quinone MK-8(H2), the presence of mycolic acids and cell-wall chemotype IV were also consistent with the properties of members of the genus Rhodococcus. The DNA G+C content of strain H004T was 65.4 mol%. The results of DNA-DNA hybridization analyses with the closest relatives, in combination with the alkB-based phylogenetic analysis, as well as the chemotaxonomic and physiological data, demonstrated that isolate H004T represents a novel species of the genus Rhodococcus, for which the name Rhodococcus sovatensissp. nov. is proposed. The type strain is H004T (=DSM 102881T=NCAIM B.02632T).

Rufibacter quisquiliarum sp. nov., a new member of the phylum Bacteroidetes isolated from a bioreactor treating landfill leachate.

A novel bacterium, CAI-18bT, was isolated from a bioreactor that treated landfill leachate using an oligotrophic growth medium. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CAI-18bT is a member of the genus Rufibacter, showing 97.1 % pairwise similarity to Rufibacter roseus H359T, 96.4 % to Rufibacter tibetensis 1351T, 96.4 % to Rufibacter glacialis MDT1-10-3T and 96.0 % to Rufibacter immobilis MCC P1T. Strain CAI-18bT was rod-shaped, motile, oxidase- and catalase-positive. The predominant fatty acids were iso-C15 : 0 (24.1 %) and iso-C17 : 1 I (22.3 %), the major respiratory quinone was MK-7, and the predominant polar lipids were phosphatidylethanolamine and an unknown aminophospholipid. The G+C content of the genomic DNA of strain CAI-18bT was 50.7 mol%. The novel bacterium can be distinguished from related type strains based on its ability to assimilate N-acetylglucosamine and gentiobiose. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CAI-18bT represents a novel species of the genus Rufibacter, for which the name Rufibacter quisquiliarum sp. nov. is proposed. The type strain is CAI-18bT (=DSM 29854T=NCAIM B.02614T).

The detection and phylogenetic analysis of the alkane 1-monooxygenase gene of members of the genus Rhodococcus.

Naturally occurring and anthropogenic petroleum hydrocarbons are potential carbon sources for many bacteria. The AlkB-related alkane hydroxylases, which are integral membrane non-heme iron enzymes, play a key role in the microbial degradation of many of these hydrocarbons. Several members of the genus Rhodococcus are well-known alkane degraders and are known to harbor multiple alkB genes encoding for different alkane 1-monooxygenases. In the present study, 48 Rhodococcus strains, representing 35 species of the genus, were investigated to find out whether there was a dominant type of alkB gene widespread among species of the genus that could be used as a phylogenetic marker. Phylogenetic analysis of rhodococcal alkB gene sequences indicated that a certain type of alkB gene was present in almost every member of the genus Rhodococcus. These alkB genes were common in a unique nucleotide sequence stretch absent from other types of rhodococcal alkB genes that encoded a conserved amino acid motif: WLG(I/V/L)D(G/D)GL. The sequence identity of the targeted alkB gene in Rhodococcus ranged from 78.5 to 99.2% and showed higher nucleotide sequence variation at the inter-species level compared to the 16S rRNA gene (93.9-99.8%). The results indicated that the alkB gene type investigated might be applicable for: (i) differentiating closely related Rhodococcus species, (ii) properly assigning environmental isolates to existing Rhodococcus species, and finally (iii) assessing whether a new Rhodococcus isolate represents a novel species of the genus.

Vertical physico-chemical gradients with distinct microbial communities in the hypersaline and heliothermal Lake Ursu (Sovata, Romania).

The effect of vertical physico-chemical stratification on the planktonic microbial community composition of the deep, hypersaline and heliothermal Lake Ursu (Sovata, Romania) was examined in this study. On site and laboratory measurements were performed to determine the physical and chemical variables of the lake water, and culture-based and cultivation-independent techniques were applied to identify the members of microbial communities. The surface of the lake was characterized by a low salinity water layer while the deepest region was extremely saline (up to 300 g/L salinity). Many parameters (e.g. photosynthetically active radiation, dissolved oxygen concentration, pH, redox potential) changed dramatically from 2 to 4 m below the water surface in conjunction with the increasing salinity values. The water temperature reached a maximum at this depth. At around 3 m depth, there was a water layer with high (bacterio) chlorophyll content dominated by Prosthecochloris vibrioformis, a phototrophic green sulfur bacterium. Characteristic microbial communities with various prokaryotic taxa were identified along the different environmental parameters present in the different water layers. Some of these bacteria were known to be heterotrophic and therefore may be involved in the decomposition of lake organic material (e.g. Halomonas, Idiomarina and Pseudoalteromonas) while others in the transformation of sulfur compounds (e.g. Prosthecochloris). Eukaryotic microorganisms identified by molecular methods in the lake water belonged to genera of green algae (Mantionella and Picochlorum), and were restricted mainly to the upper layers.

Remarkable impact of PAHs and TPHs on the richness and diversity of bacterial species in surface soils exposed to long-term hydrocarbon pollution.

Nowadays, because of substantial use of petroleum-derived fuels the number and extension of hydrocarbon polluted terrestrial ecosystems is in growth worldwide. In remediation of aforementioned sites bioremediation still tends to be an innovative, environmentally attractive technology. Although huge amount of information is available concerning the hydrocarbon degradation potential of cultivable hydrocarbonoclastic bacteria little is known about the in situ long-term effects of petroleum derived compounds on the structure of soil microbiota. Therefore, in this study our aim was to determine the long-term impact of total petroleum hydrocarbons (TPHs), volatile petroleum hydrocarbons (VPHs), total alkyl benzenes (TABs) as well as of polycyclic aromatic hydrocarbons (PAHs) on the structure of bacterial communities of four different contaminated soil samples. Our results indicated that a very high amount of TPH affected positively the diversity of hydrocarbonoclastic bacteria. This finding was supported by the occurrence of representatives of the α-, ß-, γ-Proteobacteria, Actinobacteria, Flavobacteriia and Bacilli classes. High concentration of VPHs and TABs contributed to the predominance of actinobacterial isolates. In PAH impacted samples the concentration of PAHs negatively correlated with the diversity of bacterial species. Heavily PAH polluted soil samples were mainly inhabited by the representatives of the ß-, γ-Proteobacteria (overwhelming dominance of Pseudomonas sp.) and Actinobacteria.

Phylogenetic diversity of bacterial and archaeal communities inhabiting the saline Lake Red located in Sovata, Romania.

Lake Red is one of the saline lakes which were formed as a consequence of salt massif dissolution at the foot of the Gurghiu Mountains (Central Romania) at the end of the nineteenth century. The lake water had approximately 15 % w/v salt content. Phylogenetic diversity of prokaryotes inhabiting the water and sediment of the lake was studied using cultivation and cultivation-independent methods following a sampling in spring 2009. According to the results of 16S rRNA gene-based denaturing gradient gel electrophoresis (DGGE), the richness of Bacteria was higher than Archaea on the basis of the number and position of dominant bands in the gel. Sequences from DGGE bands were affiliated with Gammaproteobacteria (Halomonas and Alkalilimnicola) and Bacteroidetes (Psychroflexus) as well as Euryarchaeota. Cultivation from five different saline media resulted in 101 bacterial strains of which Gammaproteobacteria (Halomonas, Marinobacter and Salinivibrio) were the most abundant. Firmicutes (Bacillus) and Alphaproteobacteria (Aurantimonas and Roseovarius) were also identified among the isolated strains. The 16S rRNA genes from 82 bacterial and 95 archaeal clones were also phylogenetically analyzed. Bacterial clones were related to various genera of Gammaproteobacteria (Alkalilimnicola, Alkalispirillum, Arhodomonas, Halomonas, Saccharospirillum), Bacteroidetes (Gracilimonas, Psychroflexus) and Alphaproteobacteria (Oceanicola, Roseinatronobacter, Roseovarius). All of the archaeal clones sequenced corresponded to a homologous cluster affiliated with Halopelagius.