Whole-cell Fatty Acid Methyl Ester Research Articles

Apibacter muscae sp. nov., a novel bacterial species isolated from house flies.

We describe the isolation and characterization of three bacterial isolates from the common house fly, Musca domestica, caught in Londerzeel, Belgium and Huye District, Rwanda. Although isolated from distinct geographical locations, the strains show >99 % identical 16S rRNA gene sequences and are <95 % identical to type strains of Apibacter species. Whole-genome sequences were obtained for all three strains. The genomes are 2.4-2.5 Mb with a G+C content of ~30.3 mol%. Bacteriological and biochemical analysis of the strains demonstrate distinctly different characteristics compared to known Apibacter species. Particularly, the three strains investigated in this study can be distinguished from the known Apibacter species (Apibacter mensalisand Apibacter adventoris) through urease and β-glucosidase activities. Whole-cell fatty acid methyl ester analysis shows that the fatty acid composition of the novel strains is also unique. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify these isolates as representatives of a novel species of the genus Apibacter, Apibacter muscae sp. nov., in reference to its prevalence in house flies, with strain G8T (=LMG 30898T=DSM 107922T) as the type strain.

Molecular-based analysis of the bacterial community structure of spoilt latex paints in Nigeria

Biodeteriorated paints showing discolouration, loss of viscosity and foul odour have resulted in substantial economic losses in Nigeria. The present study focused on enumeration and identification of bacteria isolated from spoilt paints made in Nigeria using a non-selective nutrient agar (NA) and a semi-selective Pseudomonas Isolation Agar (PIA). The bacterial population density in a fresh paint sample was 3.0 x 103 CFU/ml while that of five samples of spoilt paint ranged from 1.0 x 103 to 1.2 x 106 CFU/ml on NA. The fresh paint had no growth on PIA while spoilt paint bacterial counts ranged from 2.0 x 103 to 8.0 x 105 CFU/ml. The most diverse colony types from the spoilt paint samples were randomly selected for purification. We subjected 9 isolates to sequencing of a portion of the 16S rRNA gene and 14 isolates to whole-cell fatty acid methyl ester profiling using the MIDI-FAME system. The most prevalently identified were Klebsiela and Providencia species. A subset of isolates were examined by transmission electron microscopy. Other detected genera included Bacillus, Brevibacillus, Lysinibacillus, and the Nocardioforms. Our results show that spoilt latex paints made in Nigeria can harbour diverse bacterial species which can cause biodeterioration.

Apibacter mensalis sp. nov.: a rare member of the bumblebee gut microbiota.

Isolates LMG 28357T (=R-53146T) and LMG 28623 were obtained from gut samples of Bombus lapidarius bumblebees caught in Ghent, Belgium. They had identical 16S rRNA gene sequences which were 95.7 % identical to that of Apibacter adventoris wkB301T, a member of the family Flavobacteriaceae. Both isolates had highly similar matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS and randomly amplified polymorphic DNA (RAPD) profiles. A draft genome sequence was obtained for strain LMG 28357T (Gold ID Gp0108260); its DNA G+C content was 30.4%, which is within the range reported for members of the family Flavobacteriaceae (27 to 56 mol%) and which is similar to that of the type strain of A. adventoris (29.0 mol%). Whole-cell fatty acid methyl ester analysis of strain LMG 28357T revealed many branched-chain fatty acids, a typical characteristic of bacteria of the family Flavobacteriaceae and a profile that was similar to that reported for A. adventoris wkB301T. MK6 was the major respiratory quinone, again conforming to bacteria of the family Flavobacteriaceae. The isolates LMG 28357T and LMG 28623 could be distinguished from A. adventoris strains through their oxidase activity. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify both isolates as representatives of a novel species of the genus Apibacter, Apibacter mensalis sp. nov., with LMG 28357T (=DSM 100903T=R-53146T) as the type strain.

Molecular Cloning and Sequencing of AlkalophilicCellulosimicrobium cellulans CKMX1 Xylanase Gene Isolated from Mushroom Compost and Characterization of the Gene Product

A xylanolytic bacterium was isolated from mushroom compost by using enrichment technique. Results from the metabolic fingerprinting, whole-cell fatty acids methyl ester analysis and 16S rDNA sequencing suggested the bacterium to be Cellulosimicrobium cellulans CKMX1. Due to the xylanolytic activity of this bacterium, isolation and characterization of the xylanase gene were attempted. A distinct fragment of about 1671 bp was successfully amplified using PCR and cloned into Escherichia coli DH5α. A BLAST search confirmed that the DNA sequence from the amplified fragment was endo-1, 4-beta-xylanase, which was a member of glycoside hydrolase family 11. It showed 98% homology withCellulosimicrobium sp. xylanase gene (Accession no. FJ859907.1) reported from the gut of Eisenia fetida in Korea. In silicophysico-chemical characterization of amino acid sequence of xylanase showed an open reading frame encoding a 556 amino acid sequence with a molecular weight of 58 kDa and theoretical isolectric point (pI) of 4.46 was computed using Expasy's ProtParam server. Secondary and homology based 3D structure of xylanase was analysed using SOPMA and Swiss-Prot software.

Modification in the properties of paper by using cellulase-free xylanase produced from alkalophilic Cellulosimicrobium cellulans CKMX1 in biobleaching of wheat straw pulp.

Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is an actinomycete that produces industrially important and environmentally safer thermostable cellulase-free xylanase, which is used in the pulp and paper industry as an alternative to the use of toxic chlorinated compounds. Strain CKMX1 was previously characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis, and 16S rDNA and was found to be C. cellulans CKMX1. Crude enzyme (1027.65 U/g DBP) produced by C. cellulans CKMX1, having pH and temperature optima of 8.0 and 60 °C, respectively, in solid state fermentation of apple pomace, was used in the production of bleached wheat straw pulp. Pretreatment with xylanase at a dose of 5 U/g after pulping decreased pulp kappa points by 1.4 as compared with the control. Prebleaching with a xylanase dose of 5 U/g pulp reduced the chlorine charge by 12.5%, increased the final brightness points by approximately 1.42% ISO, and improved the pulp strength properties. Xylanase could be substituted for alkali extraction in C-Ep-D sequence and used for treating chemically bleached pulp, resulting in bleached pulp with higher strength properties. Modification of bleached pulp with 5 U of enzyme/g increased pulp whiteness and breaking length by 1.03% and 60 m, respectively; decreased tear factor of pulp by 7.29%; increased bulk weight by 3.99%, as compared with the original pulp. Reducing sugars and UV-absorbing lignin-derived compound values were considerably higher in xylanase-treated samples. Cellulosimicrobium cellulans CKMX1 has a potential application in the pulp and paper industries.

Purification and characterization of cellulase-free low molecular weight endo β-1,4 xylanase from an alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost.

Alkalophilic Cellulosimicrobium cellulans CKMX1 isolated from mushroom compost is first report on actinomycetes that has the ability to produce thermostable cellulase-free xylanase, which is an important industrial enzyme used in the pulp and paper industry. Strain CKMX1 was characterized by metabolic fingerprinting, whole-cell fatty acids methyl ester analysis and 16Sr DNA and found to be C. cellulans CKMX1.The enzyme was purified by gel permeation and anion exchange chromatography and had a molecular mass of 29 kDa. Xylanase activity was optimum at pH 8.0 and 55 °C. The enzyme was somewhat thermostable, retaining 50 % of the original activity after incubation at 50 °C for 30 min. The xylanase had K m and V max values of 2.64 mg/ml and 2,000 µmol/min/mg protein in oat spelt xylan, respectively. All metal ions except HgCl2, CoCl2 as well as CdCl2 were well tolerated and did not adversely affect xylanase activity. The deduced internal amino acid sequence of C. cellulans CKMX1 xylanase by matrix assisted laser desorption ionization-time of flight mass spectrometry resembled the sequence of β-1,4-endoxylanase, which is a member of glycoside hydrolase family 11. Some of the novel characteristics that make this enzyme potentially effective in xylan biodegradation could be useful for pulp and paper biobleaching are discussed in this manuscript.

First Report of Sporobolomyces symmetricus Induced Red Spot Disease of Pleurotus eryngii in China.

Pleurotus eryngii is cultivated on a large scale in China and other Asian countries due to its exceptional flavor and its popularity in cuisine. In January of 2009, red spot disease on cultivated P. eryngii was observed in several workshops of mushroom producing factories located at Fangshan District, Beijing, China. Symptoms of this disease began as orange red spots 2 to 4 cm in diam. on the surface of stipes near pilei. The spots did not arise from wounding, and growth of the affected mushrooms halted. Commercial losses for these factories were estimated at 10 to 15%. Samples of diseased tissue were plated on potato dextrose agar (PDA), and dominant colonies were selected for characterization. After 3 days in YM broth (Difco) at 25°C, yeast cells developed that were ellipsoid, botuliform, 4.8 to 7.2 × 2.4 to 3.6 μm, and sediment-formed. After incubation for 1 month on YM agar at 25°C, streak cultures were nacarat, cheese-like, glistening and had a smooth surface and complete margin. On Dalmau plate cultures on cornmeal agar (Difco), no pseudohyphae were formed. The internal transcribed spacer (ITS) and D1/D2 domain of the 26S rRNA were sequenced (GenBank Accession Nos. KF314801 and KF314803). A neighbor-joining (NJ) tree for the D1/D2 domain showed that the yeast belonged to the Urediniomycetes with the highest similarity (99.6%) to Sporobolomyces symmetricus CBS 9727T. BLAST results for the ITS sequence showed that it was 100% identical to S. symmetricus CBS 9727T. Furthermore, the metabolic profile of the fungal strain also closely resembled the database profile of S. symmetricus CBS 9727T with the exception of two phenotypic characteristics (positive assimilation of D-Galactose and D-Xylose). The DNA G+C content of the strain was measured by HPLC and was found to be 55.40 mol%. The whole-cell fatty acid methyl ester major composition determined using the cells harvested from cultures grown on PDA for 3 days at 25°C was as follows: 1.23% C16:1 w7c/16:1 w6c, 21.87% C16:0, 29.40% C18:0 antei/C18:2 w6, 9c, 40.41% C18:1 w9c, 3.36% C18:1 w7c, 1.93% C18:0. After comparing the morphological, phenotypic, and molecular analyses with the type strain of S. symmetricus CBS 9727T, the strain isolated in the present study was identified as S. symmetricus (2). Pathogenicity tests for the isolate were carried out with yeast suspensions (~1 × 106 CFU/ml) grown for 72 h in PDB (Difco) cultures. Mycelia of P. eryngii were cultivated for 35 days in a plastic bottle to serve as a host source. The prepared yeast cell suspensions were directly inoculated on the surface of the young fruiting body. After 5 to 7 days, red spot symptoms identical to those described above developed on the stipes and pilei and development of the mushrooms was inhibited. Fruiting bodies in negative control bottles inoculated with PDB did not develop symptoms. Koch's postulates were thus fulfilled by re-isolating S. symmetricus that was identical to the inoculated strain from lesions on the inoculated fruiting bodies. Sporobolomyces spp. are closely phylogenetically related to the genus Rhodotorula. Pathogenic potential of Rhodotorula species has been increasingly recognized in recent years (1). To our knowledge, this is the first report of S. symmetricus-induced red spot disease of P. eryngii in China. Information on this pathogen will be useful in the development of management practices for this emerging disease in the near future.

Novel phosphate solubilizing bacteria ‘Pantoea cypripedii PS1’ along with Enterobacter aerogenes PS16 and Rhizobium ciceri enhance the growth of chickpea (Cicer arietinum L.)

Phosphate solubilizing bacteria (PSB) are known to convert the insoluble forms of phosphate to soluble one and make them available for plant uptake. The present study aimed to isolate PSB from the rhizosphere of chickpea (Cicer arietinum L. cv. GPF2) and examine their effect on the growth and seed number. The isolated PSB were analyzed for phosphate solubilization, indole acetic acid and siderophore production. PSB were characterized for phenotypic and biochemical properties, BIOLOG and whole-cell fatty acid methyl ester profile and found to be closely related to Pantoea cypripedii and Enterobacter aerogenes based on 16s rRNA gene sequencing. A high increase in growth of C. arietinum was observed when innoculated with PSB in tricalcium phosphate amended soils. A higher uptake in total P (53 %) of plants was observed when inoculated with mixture of P. cypripedii and E. aerogenes along with Rhizobium ciceri as compared to respective control plants which significantly increased the seed number (98.3 %) and seed weight (46.1 %). This study demonstrated the ability of novel PSB P. cypripedii along with E. aerogenes and R. ciceri to promote chickpea growth.

A genetic discontinuity in root-nodulating bacteria of cultivated pea in the Indian trans-Himalayas.

Evolutionary relationships of 120 root-nodulating bacteria isolated from the nodules of Pisum sativum cultivated at 22 different locations of the trans-Himalayan valleys of Lahaul and Spiti in the state of Himachal Pradesh of India were studied using 16S rRNA gene PCR-RFLP, ERIC-PCR, sequencing of 16S rRNA, atpD, recA, nodC and nifH genes, carbon-source utilization pattern (BIOLOG™), and whole-cell fatty acid profiling. The results demonstrated that all isolates belonged to Rhizobium leguminosarum symbiovar viciae (Rlv). Isolates from the two valleys were clearly separated on the basis of ERIC fingerprints, carbon-source utilization pattern, and whole-cell fatty acid methyl esters. Phylogenetic analysis of atpD, recA, nodC and nifH genes revealed a common Rlv sublineage in Spiti valley. Lahaul valley isolates were represented by three sequence types of atpD and recA genes, and four sequence types of nodC and nifH genes. Genotypes from the two valleys were completely distinct, except for two Lahaul isolates that shared nodC and nifH sequences with Spiti isolates but were otherwise more similar to other Lahaul isolates. Isolates from the two highest Spiti valley sites (above 4000 m) had a distinctive whole-cell fatty acid profile. Spiti valley isolates are closely related to Rlv sublineages from Xinjiang and Shanxi provinces in China, while Lahaul valley isolates resemble cosmopolitan strains of the western world. The high mountain pass between these valleys represents a boundary between two distinct microbial populations.

Phaeobacter caeruleus sp. nov., a blue-coloured, colony-forming bacterium isolated from a marine electroactive biofilm

Three isolates (LMG 24369(T), LMG 24370 and R-26156) obtained from a marine electroactive biofilm that was grown on a cathodically polarized electrode were investigated by using a polyphasic taxonomic approach. Whole-cell fatty acid methyl ester and 16S rRNA gene sequence analyses indicated that the isolates were members of the genus Phaeobacter, class Alphaproteobacteria. Genotypic and phenotypic analyses demonstrated that the three isolates represent a novel species of the genus Phaeobacter, for which the name Phaeobacter caeruleus sp. nov. is proposed. The type strain is LMG 24369(T) (=CCUG 55859(T)). The DNA G+C content of strain LMG 24369(T) is 63.6 mol%.

Nautella italica gen. nov., sp. nov., isolated from a marine electroactive biofilm

Five isolates obtained from a marine electroactive biofilm grown on a stainless steel cathode were investigated by using a polyphasic taxonomic approach. Analyses of whole-cell fatty acid methyl esters and 16S rRNA gene sequences showed that the isolates belonged to the Roseobacter lineage of the class Alphaproteobacteria. Both phenotypic and genotypic analyses demonstrated that the five new isolates constituted a single species that did not represent a recognized member of the Roseobacter lineage. Therefore the five isolates represent a novel genus and species, for which the name Nautella italica gen. nov., sp. nov. is proposed. The type strain is LMG 24365(T) (=CCUG 55857(T)). The DNA G+C content of the type strain is 61 mol%.

Ruegeria scottomollicae sp. nov., isolated from a marine electroactive biofilm

Seventy isolates were obtained from a marine electroactive biofilm that was generated on a cathodically polarized stainless steel electrode (Genoa, Italy). The genetic diversity was investigated by means of BOX-PCR fingerprinting and two clusters of isolates with similar BOX-PCR profiles were delineated. Whole-cell fatty acid methyl ester analysis and 16S rRNA gene sequence analysis showed that the isolates belonged to the Roseobacter lineage of the class Alphaproteobacteria. DNA-DNA hybridization experiments and a biochemical analysis demonstrated that four isolates belonged to the species Ruegeria mobilis. However, 66 isolates from the second BOX-PCR cluster constituted a novel species within the genus Ruegeria, for which the name Ruegeria scottomollicae sp. nov. is proposed. The DNA G+C content was 61.0+/-0.4 %. The type strain is LMG 24367(T) (=CCUG 55858(T)).

Alteromonas genovensis sp. nov., isolated from a marine electroactive biofilm and emended description of Alteromonas macleodii Baumann et al. 1972 (Approved Lists 1980)

Thirty-five isolates obtained from a marine electroactive biofilm grown on a stainless steel cathode (Genoa, Italy) were investigated by a polyphasic taxonomic approach. Whole-cell fatty acid methyl ester analysis indicated that the isolates belonged to the class Gammaproteobacteria and were closely related or belonging to the genus Alteromonas. Genomic fingerprinting using the BOX-PCR primer delineated five clusters of isolates with similar BOX-PCR fingerprints. This study demonstrated that isolates from four of the BOX-PCR clusters belonged to Alteromonas macleodii and that the 14 isolates representing BOX-PCR cluster 1 constituted a novel species, which shared 98.4 % 16S rRNA gene sequence similarity with its nearest phylogenetic neighbour, Alteromonas hispanica. Both phenotypic and genotypic analyses enabled this novel species, for which the name Alteromonas genovensis sp. nov. is proposed, to be differentiated from established species of the genus Alteromonas. The DNA G+C content of Alteromonas genovensis sp. nov. is 44.5 mol% and the type strain is LMG 24078(T) (=CCUG 55340(T)).

Microbial community in the rhizosphere of young maize seedlings is susceptible to the impact of introduced pseudomonads as indicated by FAME analysis

Two species of Pseudomonas (i.e. P. chlororaphis or P. putida) derived from a maize rhizosphere were studied for their impact on the structure of the microbial community in the rhizosphere of young maize seedlings after inoculation. The culturable bacteria and total microbial communities were analyzed based on profiles of whole-cell fatty acid methyl esters (MIDI-FAME). The introduction of Pseudomonas species resulted in the shift from the Gram-positive dominated culturable community in the rhizosphere of uninoculated maize to more Gram-negative populations in the rhizospheres of the inoculated plants. For the total rhizosphere communities, 43, 47 and 42 FAMEs were detected in the uninoculated maize and the samples inoculated with P. chlororaphis or P. putida, respectively. In contrast to the culturable communities, low concentrations of marker FAMEs for Gram-positives (i15:0, a15:0, i16:0) were found in the profiles of the total rhizosphere communities. The maize inoculations resulted in an enrichment of some Gram-negative isolates; however, Gram-positive bacteria, Cytophaga/Flavobacterium and saprophytic fungi were found in the uninoculated rhizosphere.

Characterization of Phosphate-Solubilizing Fluorescent Pseudomonads from the Rhizosphere of Seabuckthorn Growing in the Cold Deserts of Himalayas

Isolation and characterization of fluorescent pseudomonads with high phosphate-solubilizing ability is reported from the alkaline and calcium-rich soils with low P availability in the cold desert region of Lahaul and Spiti in the trans-Himalayas of India. Of 216 phosphate-solubilizing isolates, 12 exhibiting high solubilization of tricalcium phosphate (TCP) in NBRIP liquid culture were identified as Pseudomonas trivialis, P. poae, P. fluorescens, and Pseudomonas spp. on the basis of phenotypic features, whole-cell fatty acids methyl ester (FAME) profiles, and 16S rDNA sequencing. These isolates also showed relatively high solubilization of North Carolina rock phosphate (NCRP) in comparison to the solubilization of Mussoorie rock phosphate (MRP) and Udaipur rock phosphate (URP). The solubilization of phosphate substrates by P. trivialis and P. poae is reported for the first time.

Phenotypic characterization of Escherichia coli through whole-cell fatty acid profiling to investigate host specificity

The objective of the study was to investigate whole-cell fatty acid methyl ester (FAME) profiles of 605 Escherichia coli isolates to determine their host specificity. The isolates were cultured from six possible sources of fecal pollution; 180 isolates from sewage, 85 from dairy cow, 98 from chicken, 76 from swine, 94 from deer, and 72 from waterfowl, mostly geese and ducks. The FAME profiles were presented as the relative masses of 12 FAMEs identified in the isolates and it was found that none of the six hosts carried a “signature” FAME, a FAME that is uniquely associated with a particular host category. However, two-sample t-test analyses indicated that the mean relative masses of seven FAMEs out of the 12 identified showed statistically significant differences (95% confidence interval) between isolates of human and non-human origins. In addition, a linear discriminant function based on mean relative mass variations in individual FAMEs classified the known-source isolates into their respective host categories with a 47.6% average rate of correct classification (ARCC) in a six-way discriminant analysis. The ARCC increased to 61.3% when the individual hosts were pooled into larger categories of human, livestock, and wildlife. The accuracy was 75.5% when isolates of human origin were discriminated against those of non-human origins. Random cluster formation analysis indicated that the library size was sufficient to prevent random grouping among the isolates.

Whole-Cell Fatty Acid Composition of Total Coliforms to Predict Sources of Fecal Contamination

The objective of this study was to test the hypothesis that total coliforms isolated from human and nonhuman sources have distinctly different whole-cell fatty acid methyl ester (FAME) profiles. The FAME profiles studied included total coliforms isolated from sewage; feces of livestock, including bovine (dairy cattle), poultry, and swine; and feces of wildlife, including waterfowl and deer. Multiple samples within each category were collected and 303 total coliform isolates were cultured. It was found that the FAMEs 12:0 2OH and 14:0 2OH were exclusively associated with sewage samples, whereas the FAMEs 18:0 and 19:0 ISO were identified only in isolates from the livestock samples. In addition to the presence of signature FAMEs, the average relative masses of 16:1 ω7c , 18:1 ω7c , and 19:0 CYCLO ω8c were significantly different between human and nonhuman sources of total coliforms. A linear discriminant function based on these differences discriminated total coliform isolates of human origin against the ot...

Isolation and characterization of phosphate solubilizing bacteria from the rhizosphere of crop plants of Korea

Whole-cell fatty acids methyl ester (FAME) profile and 16S rDNA sequence analysis were employed to isolate and identify the bacterial groups that actively solubilized phosphates in vitro from rhizosphere soil of various crops of Korea. Out of several hundred colonies that grew on Pikovskaya's medium 13 best isolates were selected based on the solubilization of insoluble phosphates in liquid culture and further characterized and identified. They were clustered under the genera Enterobacter, Pantoea and Klebsiella and the sequences of three representative strains were deposited in the GenBank nucleotide sequence data library under the accession numbers AY335552, AY335553, AY335554.

Characterization of esculin-positivePseudomonas fluorescens strains isolated from an underground brook

A group of sixteen esculin-positive fluorescent pseudomonads isolated from an underground brook flowing through a cave complex was characterized by biotyping, multiple enzyme restriction fragment length polymorphism analysis of 16S rDNA (MERFLP), ribotyping and whole-cell fatty-acid methyl-esters analysis (FAME). All strains were phenotypically close to Pseudomonas fluorescens, but they revealed high biochemical variability as well as some reactions atypical for P. fluorescens species. Because identification of pseudomonads by of biochemical testing is often unclear, further techniques were employed. Fingerprints obtained by MERFLP clearly showed that all strains represent P. fluorescens species. Ribotyping separated the strains analyzed into four groups corresponding almost completely (with the exception of one strain) to the clustering based on biochemical profiles. FAME analysis grouped all the strains into one cluster together with the P. putida (biotype A, B), P. chlororaphis and P. fluorescens biotype F representatives, but differentiated them from other FAME profiles of all pseudomonads included in the standard library TSBA 40 provided by MIDI, Inc.

Application of temperature-gradient gel electrophoresis in taxonomy of coryneform bacteria.

Strains belonging to the Gram-positive coryneform soil bacteria were screened genotypically by temperature-gradient gel electrophoresis (TGGE). This method allows the sequence-specific separation of amplified fragments of 16S rRNA genes. A total of 115 reference strains representing the majority of the species of the genera Aeromicrobium, Agromyces, Arthrobacter, Aureobacterium, Cellulomonas, Curtobacterium, Nocardioides and Terrabacter were characterized. Depending on the genus investigated, the resolution limit of the technique appeared to be at the species or genus level or intermediate between the two. Aberrant TGGE profiles of strains within particular taxa revealed genomic heterogeneity and generic misclassification of nine strains studied. Beyond that, indications of 16S rRNA gene heterogeneity were found within the genomes of three Curtobacterium strains. The misclassifications revealed by TGGE were confirmed using whole-cell fatty acid methyl ester analysis and subsequent comparison with a database. TGGE has been demonstrated to be a useful tool in bacterial taxonomy.