ALBERT

Description: During characterization of autochthonic Vyhna travertine source microflora, several bacterial strains were isolated and characterised. Isolate T6, a halotolerant, moderately alkaliphilic and thermophilic bacterial isolate, was further characterised based on physiological, microbiological and biochemical tests and phylogenetic 16S rRNA analysis. On the basis of the results obtained, the T6 isolate should be placed in the genus Oceanobacillus , and it is probably a prototype of a novel bacterial species. Characterization of the T6 isolate broadens our knowledge on variability of halophilic bacteria of Oceanobacillus genus and expands data on travertine-associated bacterial communities.

Description: Corynebacterium striatum is often dismissed as a contaminant when cultivated from blood samples; indeed, it is a skin saprophyte that may therefore be introduced into the clinical specimen accidentally. Nevertheless, the organism can be responsible for true bacteraemias, and multidrug resistance spread among nosocomial strains is of increasing concern. Specific criteria for testing have not been defined yet, but we however suggest to report clear resistances (i.e. absence of any inhibition zones with the disc test), in order to try to understand this species behaviour under antibiotic exposure. In this context, features of a blood isolate (strain DSM 45711) are here depicted.

Description: The yeast Saccharomyces cerevisiae accumulates the high levels of inorganic polyphosphates (polyPs) performing in the cells numerous functions, including phosphate and energy storage. The effects of vacuolar membrane ATPase (V-ATPase) dysfunction were studied on polyP accumulation under short-term cultivation in the P i –excess media after P i starvation. The addition of bafilomycin A1, a specific inhibitor of V-ATPase, to the medium with glucose resulted in strong inhibition of the synthesis of long-chain polyP and in substantial suppression of short-chain polyP. The addition of bafilomycin to the medium with ethanol resulted in decreased accumulation of high-molecular polyP, while the accumulation of low-molecular polyP was not affected. The levels of polyP synthesis in the mutant strain with a deletion in the vma2 gene encoding a V-ATPase subunit were significantly lower than in the parent strain in the media with glucose and with ethanol. The synthesis of the longest chain polyP was not observed in the mutant cells. The synthesis of only the low-polymer acid-soluble polyP fraction occurred in the cells of the mutant strain. However, the level of polyP1 was nearly tenfold lower than compared to the cells of the parent strain. Both bafilomycin A1 and the mutation in vacuolar ATPase subunit vma2 lead to a considerable decrease of cellular polyP accumulation. Thus, the defects in ΔμH + formation on the vacuolar membrane resulted in the decrease of polyP biosynthesis in S. cerevisiae .

Description: The Bacillus cereus sensu lato group includes potentially pathogenic bacteria that are ubiquitous in the environment and, importantly, could also be present in food products. This study focuses on emetic isolates which presumably could cause acute food poisoning and emetic syndrome. Here, we evaluate the ability of psychrotolerant Bacillus weihenstephanensis MC118 (isolated from soil) and mesophilic B. cereus BOD3/9 isolated from milk to germinate and multiply at 7 and 30 °C. Whereas the rates of germination at 30 °C in milk and nutrient broth of MC118 and BOD3/9 were similar, MC118, but not BOD3/9, proliferated to achieve relatively high numbers (∼10 6  colony-forming units/g) within 7 days of incubation at 7 °C. Mesophilic BOD3/9 showed a slight decrease of cell concentration in similar studies at 7 °C. Genotyping with repetitive extragenic palindromic sequence-based PCR and pulsed field gel electrophoresis revealed significant similarities between BOD3/9 and emetic reference B. cereus F4810/72 strain, while the B. weihenstephanensis MC118 isolate was more similar to the B. weihenstephanensis non-emetic reference DSMZ11821 strain. Our data suggest that emetic isolates that are also psychrotolerant, such as MC118, could constitute a hazard in the dairy industry, where milk could be a suitable medium for germination and growth.

Description: The ability of three psychrotrophic Gram-negative bacilli isolated from Chilean Patagonian cold freshwater rivers to produce bioactive metabolites was evaluated. The strains were isolated from cold waters rivers and identified by their biochemical properties and 16S rRNA gene analysis. The metabolites fractions showing antibacterial activity were obtained by solvent extraction and partially characterized by gas–mass chromatography (GC-MS). Antibacterial activity of the fractions was evaluated by an agar-well diffusion test upon 14 bacterial strains, both Gram positive and Gram negative. Thermal and proteolytic resistances of the antibacterial metabolites fractions were also evaluated. Molecular analysis allows the identification of the three Patagonian strains as Pseudomonas sp. RG-6 ( Pseudomonas brenneri 99.6 % identity), Pseudomonas sp. RG-8 ( Pseudomonas trivialis 99.6 % identity) and Yersinia sp. RP-3 ( Yersinia aldovae 99.5 % identity). These extracts were able to inhibit both Gram-positive and Gram-negative bacteria but not Listeria monocytogenes . The antibacterial activity of the filtrated supernatants was lost at temperatures ≥60 °C, and was not affected by proteinase K treatment. The chemical structure of the active molecule remains to be elucidated, although the GC-MS analysis of the filtrates suggests that compounds like sesquiterpenes derivatives from β-maaliene or δ-selinene could be responsible of this antibacterial activity. Pristine cold freshwater streams showed to be interesting sources of metabolites-producing microorganisms with antibacterial activity.

Description: Mucosal immunization with non-living antigens usually requires the use of an adjuvant. The adjuvant activity of Bacillus firmus in the mucosal immunization of mice was described by our laboratory previously. In the present study, subcellular localization of B . firmus activities was followed. After mechanical disintegration, subcellular components of bacterium were fractionated by differential centrifugation and salting out. Bacterial cell walls, cytoplasmic membrane fraction, soluble cytoplasmic proteins, and ribosomal fractions were isolated. Their effect on the mouse immune system was studied. Lymphocyte proliferation and immunoglobulin formation in vitro were stimulated by bacterial cell wall (BCW), cytoplasmic membrane (CMF), and ribosomal fractions. BCW and CMF increased antibody formation after intratracheal immunization of mice with influenza A and B viruses, and increased protection against subsequent infection with influenza virus. The BCW fraction even induced intersubtypic cross-protection: Mice immunized with A/California/7/04 (H3N2) + BCW were resistant to the infection by the highly pathogenic A/PR/8/34 (H1N1) virus.

Description: From 1993 to 2009, there was only one cluster of invasive meningococcal disease (IMD) reported in a community of children in the Czech Republic. This exceptional cluster that occurred in a preschool facility is the focus of this report. In response to the announcement of the disease, anti-epidemic precautions were put in place. Neisseria meningitidis isolates were delivered from local laboratories to the National Reference Laboratory for Meningococcal Infections in Prague. Phenotyping was performed there along with multilocus sequence typing. Related factors and microbiological results were analyzed retrospectively. In October 2009, three girls contracted IMD within a period of 1 week in a 42-member group in a preschool facility attached to the elementary school in Starý Plzenec-Sedlec. In relation to three cases of the disease, another 66 people were registered of which 58 underwent a microbiological examination. N. meningitidis was detected in a total of five (8.6 %) people. The National Reference Laboratory for Meningococcal Infections defined the type of the strain to be C: P1.18-1,34-2,38: F1-7: ST-467 (cc269) and penA 27. Tests showed the precise identity of all strains obtained from the three sick children and of two strains contracted through contact with the preschool facility. Despite the complete recovery of all patients with no permanent damage, the need for rapid cooperation between clinical sites, diagnostic laboratories, and epidemiologists was confirmed.

Description: Effects of different light conditions on development, growth, and secondary metabolism of three marine-derived filamentous fungi were investigated. Darkness irritated sexual development of Aspergillus glaucus HB1-19, while white, red, and blue lights improved its asexual behavior. The red and blue lights improved asexual stroma formation of Xylaria sp. (no. 2508), but the darkness and white light inhibited it. Differently, development of Halorosellinia sp. (no. 1403) turned out to be insensitive to any tested light irradiation. Upon the experimental data, no regularity was observed linking development with secondary metabolism. However, fungal growth showed inversely correlation with productions of major bioactive compounds (aspergiolide A, 1403C, and xyloketal B) from various strains. The results indicated that aspergiolide A biosynthesis favored blue light illumination, while 1403C and xyloketal B preferred red light irradiation. With the favorite light sensing conditions, productions of aspergiolide A, 1403C, and xyloketal B were enhanced by 32.9, 21.9, and 30.8 % compared with those in the dark, respectively. The phylogenetic analysis comparing the light-responding proteins of A. glaucus HB 1-19 with those in other systems indicated that A. glaucus HB 1-19 was closely related to Aspergillus spp. especially A. nidulans in spite of its role of marine-derived fungus. It indicated that marine fungi might conserve its light response system when adapting the marine environment. This work also offers useful information for process optimization involving light regulation on growth and metabolism for drug candidate production from light-sensitive marine fungi.

Description: Medicago truncatula represents a model plant species for understanding legume–bacteria interactions. M. truncatula roots form a specific root–nodule symbiosis with the nitrogen-fixing bacterium Sinorhizobium meliloti . Symbiotic nitrogen fixation generates high iron (Fe) demands for bacterial nitrogenase holoenzyme and plant leghemoglobin proteins. Leguminous plants acquire Fe via “Strategy I,” which includes mechanisms such as rhizosphere acidification and enhanced ferric reductase activity. In the present work, we analyzed the effect of S. meliloti volatile organic compounds (VOCs) on the Fe-uptake mechanisms of M. truncatula seedlings under Fe-deficient and Fe-rich conditions. Axenic cultures showed that both plant and bacterium modified VOC synthesis in the presence of the respective symbiotic partner. Importantly, in both Fe-rich and -deficient experiments, bacterial VOCs increased the generation of plant biomass, rhizosphere acidification, ferric reductase activity, and chlorophyll content in plants. On the basis of our results, we propose that M. truncatula perceives its symbiont through VOC emissions, and in response, increases Fe-uptake mechanisms to facilitate symbiosis.

Description: Helicobacter pylori was examined in 110 patients (82 (74.5) with gastritis, 18 (16.4) with duodenitis, six (5.5) with duodenal ulcer and gastroesophageal reflux, and four (3.6 %) with normal) with gastrointestinal problems living in rural area, no history of macrolide use, and detected by culture (71.8) or direct detection from gastric biopsies by PCR (82.7 %). Also, cagA gene was identified using PCR and was found positive in 68/91 (74.7 %) strains. The prevalence of clarithromycin-resistant H. pylori was investigated by two methods including PCR–RFLP (7.7 (A2142G 1.1 and A2143G 6.6 %)) and twofold agar dilution (8.9 %) to detect phenotypic and genotypic status simultaneously. Among all the H. pylori positive patients, eight (8.8 %) isolates were found to be resistant to clarithromycin by at least one of the AD and/or PCR–RFLP methods . H. pylori positive rates were significantly correlated with patients' sex, age, and endoscopic findings ( p  = 0.040, 〈0.001 and 〈0.001, respectively). There were no differences in gender or endoscopic findings related to cagA + and cagA − patients. The gene of cagA was not significantly helpful in predicting the clinical outcome of H. pylori infection alone. In conclusion, we revealed that there was a low prevalence of primer clarithromycin resistance in patients living in rural area with no history of macrolide use. The prevalence of mutant strains among the macrolide-resistant H. pylori varies even geographically between close provinces.

Description: Metal-related genes ( afe _ 0654 , afe _ 0671 , afe _ 0674 , afe _ 1143 , afe _ 1144 , and afe _ 2126 ) were cloned to identify whether those genes existed in Acidithiobacillus ferrooxidans strain DC ( A . ferrooxidans DC). The deduced amino acid sequences of those genes were analyzed by bioinformatics. The tolerance levels of A . ferrooxidans DC to Mn 2+ , Zn 2+ , and Cd 2+ were determined, which were 0.52, 0.42, and 0.16 mol/L for ferrous iron-grown cells and 0.38, 0.18, and 0.08 mol/L for sulfur-grown cells, respectively. Real-time quantitative PCR was employed to analyze the transcriptional levels of the metal-related genes when ferrous iron- and sulfur-grown cells of A . ferrooxidans DC, respectively, exposed to Mn 2+ , Zn 2+ , and Cd 2+ . The metal-related genes were up-regulated when A . ferrooxidans DC exposed to Mn 2+ . When A . ferrooxidans DC exposed to Zn 2+ , the metal-related genes were up-regulated in sulfur-grown cells; afe _ 0654 and afe _ 0674 were down-regulated, and the others were up-regulated in ferrous iron-grown cells. Afe _ 2126 was down-regulated, and the others were up-regulated when A . ferrooxidans DC exposed to Cd 2+ . According to experimental results and bioinformatics analysis, the proteins encoded by afe _ 0654 and afe _ 0674 may relate with Mn 2+ and Cd 2+ efflux. It needed further study whether they relate with Zn 2+ transport. Proteins encoded by afe _ 0671 , afe _ 1143 , and afe _ 1144 may relate with the efflux of Mn 2+ , Zn 2+ , and Cd 2+ . The protein encoded by afe _ 2126 may relate with Mn 2+ and Zn 2+ efflux and Cd 2+ uptake.

Description: Bacteria-assisted bioremediation is widely recognized as a low-cost method to minimize the consequences of soil pollution with toxic metals originating from industrial sites. Strains used in bioremediation have to deal with high metal load via biosorption, reduction, bioprecipitation, metal sequestration, and/or chelation. Actinobacteria, and streptomycetes in particular, are considered a perspective group for bioremediation as natural soil inhabitants with extensive secondary metabolism. Nevertheless, there is no reference information on survival of the model streptomycetes in the presence of the most abundant metal pollutants. Also, there are no reports describing the selection approaches towards improvement of bioremediation properties. In this work, the resistance of Streptomyces coelicolor M145 and Streptomyces sioyaensis Lv81 to certain transition metals and their growth under different pH values are described for the first time. Spontaneous chromate-resistant S. sioyaensis Lv81-138 strain was selected in the course of this work. Strain Lv81-138 is the most efficient actinobacterial Cr(VI) reducer reported so far, capable of converting 12 mmol/L of Cr(VI) into Cr(III) in a medium supplemented with 50 mmol/L K 2 CrO 4 .

Description: Endophytic fungal communities in leaves of deciduous trees usually undergo pronounced seasonal changes. We hypothesised that such compositional shifts are predominantly caused by annuality of the leaves and therefore less pronounced in fungi colonising the perennial substrates bark and corticolous lichens. To test this hypothesis, thalli of the foliose lichen-forming fungal species Xanthoria parietina and Physconia distorta , along with the adjacent bark, were sampled during spring and autumn at two sides of a single tree in southern Germany. Analysis of clone libraries by restriction fragment length polymorphism (RFLP) revealed 588 singleton and 221 non-singleton RFLP-types of non-lichenised fungi. The communities differed significantly between host lichen species. Season and exposure had only a significant impact when the two factors were combined in the analysis. Accordingly, bark- and/or the lichen-associated fungal communities change throughout the year’s course, a finding that rejects the initial hypothesis. This survey revealed valuable information for future broad-based studies, by indicating that a relatively high diversity of non-lichenised fungi is associated with corticolous lichen thalli and the adjacent bark. Furthermore, the structure of non-lichenised fungal assemblages associated with corticolous lichen communities obviously depends at least on the following factors: ‘lichen species’, ‘exposure’, and ‘season’.

Description: A monoclonal antibody (MAb) was generated against the capsid protein (ORF 72) of koi herpesvirus (KHV) isolated from diseased koi Cyprinus carpio in Taiwan. The clone of MAb-B2 was obtained by immunizing mice with whole virus particles and further identified using indirect enzyme-linked immunosorbent assay and Western blot assay. In addition, it detected KHV in KHV-infected cells but not in those of mock-infected cells as demonstrated by indirect immunofluorescence assay. The neutralization test showed that MAb-B2 neutralized KHV. Furthermore, we uncovered that MAb-B2 recognizes the ORF72 of KHV as revealed by liquid chromatography–tandem mass spectrometry and Western blot assays. Additionally, MAb-B2 has been used as a diagnostic tool for detection of KHV in clinical samples by immunohistochemistry. Collectively, our results indicated that MAb-B2 could be used in the development of a diagnostic kit for diagnosis of KHV infections and ORF72 protein of KHV might be a candidate for future vaccine development.

Description: Murein polysaccharides may contribute to a considerable part of the dry matter of bacterial cells. Their utilization by protozoa inhabiting the rumen is, however, poorly recognized. The objective of this study was to examine the ability of three species of ciliates, i.e., Eudiplodinium maggii , Diploplastron affine , and Entodinium caudatum of digest, and ferment these saccharides. The cultivation experiments showed that the enrichment of growth medium with bacterial cell wall β-glycans increased the ciliate number ( p  〈 0.05). A statistically significant increase ( p  〈 0.01) was followed by a continuous decrease ( p  〈 0.01) in the percentage of individuals containing β-glycans particles after 4- and 24-h incubation of ciliates with this substrate, respectively. The enzymatic experiments confirmed the ability of the examined protozoa to digest murein. E. caudatum exhibited the highest activity (8.2 unit (U)/mg protein per min), and E. maggii , the lowest (3.0 U/mg protein per min). The production rates of volatile fatty acids by starved and fed ciliate species were 0.7 and 1.6 ( E. caudatum ) pmol/ciliate cell per h, 30.5 and 42.5 ( E. maggii ) pmol/ciliate cell per h, and 8.3 and 19.2 ( D. affine ) pmol/ciliate cell per h ( p  〈 0.05).

Description: The aim of this investigation was to determine the persistence of biofilm-associated antibiotic resistance developed by methicillin-sensitive Staphylococcus aureus (MSSA), of different capsular types, during biofilm formation. Because of superiority of the tissue culture plate (TCP) over the Congo Red Agar (CRA) method for measuring biofilm formation, it was used to determine the persistence of the antibiotic resistance developed by the isolates in biofilms. The antibiotic resistance was found to persist for 3–4 wk post-propagation as planktonic subcultures. Interestingly, some strains even developed resistance to vancomycin and/or teicoplanin. However, no association of either biofilm formation or persistent antibiotic resistance with the major capsular phenotype was observed. These observations highlight the potential significance of (a) determining the antibiograms of S . aureus subcultured from biofilms developed in vitro using the TCP method as well as from planktonic cultures for formulation of an optimal therapeutic strategy, and (b) continuing to identify predominant non-capsular antigens contributing to biofilm formation, regardless of the capsular phenotype for the development of an effective potentially broad-spectrum vaccine for prevention of bovine mastitis caused by S . aureus .

Description: In the present study, we expressed the chiA74 gene of Bacillus thuringiensis in Escherichia coli K12 and demonstrated that the active ChiA74 enzyme was produced at a high level in this strain. The ChiA74 enzymatic activity (in units per milliliter) was approximately 500 % greater in E. coli K12 when compared to that produced in E. coli DH5α. Moreover, we showed that, when using our protocol, ChiA74 preparations obtained from recombinant E. coli K12 did not contain live bacteria, although transformable DNA ( erm , bla genes) was detected. Nucleic acids were subsequently easily eliminated when samples were treated with magnesium. Importantly, ChiA74 was secreted by E. coli K12 and the active enzyme was shown to generate chitin-derived oligosaccharides (C-OGS) with degrees of polymerization of 2, 3, 4, 5, and 6. From an applied perspective, the C-OGS showed activity against various pathogenic bacteria. In addition, we demonstrated that ChiA74 was not toxic to Hek 293 and 3T3 L1 cells, i.e., the enzyme did not induce apoptosis or affect normal cellular cycle and also did not produce abnormal changes in cell morphology. The potential biotechnological use of producing endochitinase of B. thuringiensis in a microorganism recognized as safe (i.e., E. coli K12) is discussed.

Description: Herpesvirus infections, such as those induced by human cytomegalovirus (HCMV), induce specific DNA damages. DNA damages can lead to cell mutation, death, apoptosis and immune system activation. Various types of DNA damage are repaired through multiple repair pathways, such as base excision, nucleotide excision, homologous recombination and nonhomologous end joining. Changes in the activity of DNA repair proteins during viral infection can cause disturbances in the DNA repair system and change its mechanisms. This report reviews results from studies, assaying a DNA repair system in HCMV infection.

Description: Candida parapsilosis produces secreted aspartic proteinases (Saps), which contribute to the virulence of this opportunistic pathogen. Gene family containing as many as 14 sequences potentially encoding secreted aspartic proteinases was identified in C. parapsilosis genome. Of them, SAPP1 and SAPP2 genes have been extensively characterized, but only now do we report that two SAPP2 homologs sharing 91.5 % identity occur in C. parapsilosis genome. Existence of SAPP2 homologs points to unexpected complexity of the SAPP gene family.

Description:    The objective of this study was to assess the genotypic diversity associated with antimicrobial susceptibility of Salmonella serovars isolated from patients arriving with diarrhoea to six hospitals of Tehran, Iran. During 2007–2008, a cross-sectional convenience study was performed. Stool samples were screened for the presence of Salmonella , serotyped, tested for antimicrobial susceptibility using disk diffusion and examined for the presence of relevant resistance genes and integrons by PCR. A total of 1,120 patients were screened for the presence of Salmonella . Out of 71 Salmonella isolates recovered, the following serovars were identified: 17 Typhi, 14 Paratyphi C, 13 Enteritidis, 11 Paratyphi B, 10 Paratyphi A and six Infantis. Most resistance was observed towards sulfamethoxazole (30%), tetracyclines (25%), nalidixic acid (22%), spectinomycin (17%), trimethoprim (15%), ampicillin (14%) and kanamycin (14%). The tetracycline resistance genes tet (A), tet (B), and tet (G) were found in 28%, 14% and 6% of the tetracycline resistant isolates, respectively. The genes aadA , aadB , strA , strB and aphA1-Iab were present in 83%, 55%, 34%, 1% and 17% of the aminoglycoside resistant isolates, respectively. Additionally, bla PSE and bla TEM β-lactamase genes were detected in 63% and 18% of the ampicillin-resistant isolates. The 23 sulphonamide resistant isolates harboured sul1 and intI1 genes, typical to class 1 integrons. Nine of these isolates also yielded amplicons for intI2 (class 2 integrons). The presence of multi-drug resistant Salmonella may compromise the successful treatment of enteric infection diseases. The enforcement of strict prescription practices will help to minimise the emergence of resistance. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0099-4 Authors Mercedeh Tajbakhsh, Research Center for Gastroenterology and Liver Disease, Shahid Beheshti University of Medical Science, Tehran, Iran Rene S. Hendriksen, National Food Institute, Technical University of Denmark, Kgs. Lyngby, Denmark Zahra Nochi, Research Center for Gastroenterology and Liver Disease, Shahid Beheshti University of Medical Science, Tehran, Iran Mohammad Reza Zali, Research Center for Gastroenterology and Liver Disease, Shahid Beheshti University of Medical Science, Tehran, Iran Frank M. Aarestrup, National Food Institute, Technical University of Denmark, Kgs. Lyngby, Denmark Lourdes Garcia-Migura, Centre de Recerca en Sanitat Animal (CReSA), Campus UAB, edifici CReSA, 08193 Bellaterra, Barcelona, Spain Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The antifungal activities of 85 lactic acid bacteria strains isolated from fermented wax gourd against the four fungal species, Penicillium oxalicum , Aspergillus flavus , Aspergillus sydowii , and Mucor racemosus , were determined. Inhibitory activity against at least one or more fungal species was observed with 27 Weissella cibaria and 11 Weissella paramesenteroides strains. Among these strains, W. cibaria 861006 and W. paramesenteroides 860509 showed greater inhibitory activities and were therefore selected for further analysis. The results suggested that the antifungal activities were originated from the organic acids produced by W. cibaria 861006 and W. paramesenteroides 860509. The application tests indicated that the growth of P. oxalicum could be effectively inhibited by W. cibaria 861006 for 6 days on grape surfaces. However, W. paramesenteroides 860509 could only remain its inhibition effect for 48 h. The findings obtained in this study suggest the potential use of W. cibaria 861006 as a bio-protective agent against fungi for agricultural purposes or ready-to-eat fresh fruit and vegetable products. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0101-1 Authors Wei-tse Lan, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan Township, Taoyuan County 333, Taiwan Yi-sheng Chen, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan Township, Taoyuan County 333, Taiwan Hui-chung Wu, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan Township, Taoyuan County 333, Taiwan Fujitoshi Yanagida, The Institute of Enology and Viticulture, Yamanashi University, 1-13-1 Kitashin, Kofu, Yamanashi 400-0005, Japan Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Eleven strains of Lactobacillus collected in the Culture Collection of Dairy Microorganisms (CCDM) were evaluated for selected probiotic properties such as survival in gastrointestinal fluids, antimicrobial activity, and competition with non-toxigenic Escherichia coli O157:H7 for adhesion on Caco-2 cells. The viable count of lactobacilli was reduced during 3-h incubation in gastric fluid followed by 3-h incubation in intestinal fluid. All strains showed antimicrobial activity and the three most effective strains inhibited the growth of at least 16 indicator strains. Antimicrobial metabolites of seven strains active against Lactobacillus and Clostridium indicator strains were found to be sensitive to proteinase K and trypsin, which indicates their proteinaceous nature. The degree of competitive inhibition of non-toxigenic E. coli O157:H7 adhesion on the surface of Caco-2 cells was strain-dependent. A significant decrease ( P  〈 0.05) in the number of non-toxigenic E . coli O157:H7 adhering to Caco-2 cells was observed with all lactobacilli. Three strains were selected for additional studies of antimicrobial activity, i.e., Lactobacillus gasseri CCDM 215, Lactobacillus acidophilus CCDM 149, and Lactobacillus helveticus CCDM 82. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0208-4 Authors Kristýna Turková, Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Purkyňova 118, 61200 Brno, Czech Republic Anja Mavrič, Institute of Dairy Science and Probiotics, Biotechnical Faculty, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Mojca Narat, Department of Animal Science, Biotechnical Faculty, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Bohuslav Rittich, Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Purkyňova 118, 61200 Brno, Czech Republic Alena Španová, Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Purkyňova 118, 61200 Brno, Czech Republic Irena Rogelj, Institute of Dairy Science and Probiotics, Biotechnical Faculty, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Bojana Bogovič Matijašić, Institute of Dairy Science and Probiotics, Biotechnical Faculty, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Adhesion of bacteria to epithelial tissue is an essential step in the progression of the urinary tract infections. Reduction of virulence factors responsible for microbial attachment may help to decrease or inhibit colonization of the host organism by pathogens. In the age of increasing bacterial antibiotic resistance, more and more attention is being paid to the use of plants and/or their bioactive components in the prevention and treatment of human infections. Asiatic acid (AA) and ursolic acid (UA), two plant secondary metabolites, were used as potential antibacterial agents. The current study aimed to determine the possible impact of AA and UA on morphology, hydrophobicity, and adhesion of clinical uropathogenic Escherichia coli strains (UPEC) to the uroepithelial cells. Our work describes for the first time the effects exerted by AA and UA on virulence factors of UPECs. The impact of both acids on the cell surface hydrophobicity of the investigated strains was very weak. The results clearly show the influence of AA and UA on the presence of P fimbriae and curli fibers, morphology of the UPECs cells and their adhesion to epithelium; however, some differences between activities of AA and UA were found. Content Type Journal Article Pages 1-8 DOI 10.1007/s12223-012-0205-7 Authors Wojnicz Dorota, Department of Biology and Medical Parasitology, Wroclaw Medical University, Mikulicza-Radeckiego 9, 50-367 Wroclaw, Poland Kicia Marta, Department of Biology and Medical Parasitology, Wroclaw Medical University, Mikulicza-Radeckiego 9, 50-367 Wroclaw, Poland Tichaczek-Goska Dorota, Department of Biology and Medical Parasitology, Wroclaw Medical University, Mikulicza-Radeckiego 9, 50-367 Wroclaw, Poland Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The phylogenetic composition, bacterial biomass, and biovolume of both planktonic and biofilm communities were studied in a low-order Bystřice stream near Olomouc City, in the Czech Republic. The aim of the study was to compare the microbial communities colonizing different biofilm substrata (stream aggregates, stream sediment, underwater tree roots, stream stones, and aquatic macrophytes) to those of free-living bacteria. The phylogenetic composition was analyzed using fluorescence in situ hybridization for main phylogenetic groups. All phylogenetic groups studied were detected in all sample types. The stream stone was the substratum where nearly all phylogenetic groups were the most abundant, while the lowest proportion to the DAPI-stained cells was found for free-living bacteria. The probe specific for the domain Bacteria detected 20.6 to 45.8 % of DAPI-stained cells while the probe specific for the domain Archaea detected 4.3 to 17.9 %. The most abundant group of Proteobacteria was Alphaproteobacteria with a mean of 14.2 %, and the least abundant was Betaproteobacteria with a mean of 11.4 %. The average value of the Cytophaga – Flavobacteria group was 10.5 %. Total cell numbers and bacterial biomass were highest in sediment and root biofilm. The value of cell biovolume was highest in stone biofilm and lowest in sediment. Overall, this study revealed relevant differences in phylogenetic composition, bacterial biomass, and biovolume between different stream biofilms and free-living bacteria. Content Type Journal Article Pages 1-9 DOI 10.1007/s12223-012-0201-y Authors Lenka Brablcová, Laboratory of Aquatic Microbial Ecology, Department of Ecology and Environmental Sciences, Faculty of Science, Palacky University, Šlechtitelů 11, 783 71 Olomouc, Czech Republic Iva Buriánková, Laboratory of Aquatic Microbial Ecology, Department of Ecology and Environmental Sciences, Faculty of Science, Palacky University, Šlechtitelů 11, 783 71 Olomouc, Czech Republic Pavlína Badurová, Laboratory of Aquatic Microbial Ecology, Department of Ecology and Environmental Sciences, Faculty of Science, Palacky University, Šlechtitelů 11, 783 71 Olomouc, Czech Republic Martin Rulík, Laboratory of Aquatic Microbial Ecology, Department of Ecology and Environmental Sciences, Faculty of Science, Palacky University, Šlechtitelů 11, 783 71 Olomouc, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    This work was aimed to develop a multiplex PCR assay to detect infectious agents such as Clavibacter michiganensis subsp. michiganensis , Fusarium sp, Leveillula taurica , and begomoviruses in tomato ( Solanum lycopersicum ) plants. Specific primer sets of each pathogen were designed based on intergenic ribosomal RNA sequences for the first three, whereas for begomoviruses, primers were designed based on conserved regions. The design also considered that the length (200–800 bp) of the PCR products was resolvable by electrophoresis; thus 296, 380, 457, and 731 bp fragments for Clavibacter , Fusarium , Leveillula , and begomoviruses, respectively, were considered. PCR conditions were optimized to amplify all the products in a single tube from genomic DNA and circumvent PCR inhibitors from infected plants. Finally, when the multiplex PCR assay was tested with tomato plants infected with any of the four pathogens, specific PCR products confirmed the presence of the pathogens. Optimized PCR multiplex allowed for the accurate and simultaneous detection of Clavibacter , Fusarium , Leveillula , and begomoviruses in infected plants or seeds from tomato. Content Type Journal Article Pages 1-8 DOI 10.1007/s12223-012-0206-6 Authors Gabriela Alejandra Quintero-Vásquez, Genetics and Molecular Biology Department, Centro de Investigaciones y Estudios Avanzados del IPN, Av IPN 2508. Delegación Gustavo A, Madero, Federal District 07360, México María Luisa Bazán-Tejeda, Genetics and Molecular Biology Department, Centro de Investigaciones y Estudios Avanzados del IPN, Av IPN 2508. Delegación Gustavo A, Madero, Federal District 07360, México Eva Martínez-Peñafiel, Genetics and Molecular Biology Department, Centro de Investigaciones y Estudios Avanzados del IPN, Av IPN 2508. Delegación Gustavo A, Madero, Federal District 07360, México Luis Kameyama-Kawabe, Genetics and Molecular Biology Department, Centro de Investigaciones y Estudios Avanzados del IPN, Av IPN 2508. Delegación Gustavo A, Madero, Federal District 07360, México Rosa María Bermúdez-Cruz, Genetics and Molecular Biology Department, Centro de Investigaciones y Estudios Avanzados del IPN, Av IPN 2508. Delegación Gustavo A, Madero, Federal District 07360, México Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Coevolution between bacteria and bacteriophages can be characterized as an infinitive constant evolutionary battle (phage-host arm race), which starts during phage adsorption and penetration into host cell, continues during phage replication inside the cells, and remains preserved also during prophage lysogeny. Bacteriophage may exist inside the bacterial cells in four forms with different evolutionary strategies: as a replicating virus during the lytic cycle, in an unstable carrier state termed pseudolysogeny, as a prophage with complete genome during the lysogeny, or as a defective cryptic prophage. Some defensive mechanisms of bacteria and virus countermeasures are characterized, and some evolutionary questions concerning phage–host relationship are discussed. Content Type Journal Article Pages 1-10 DOI 10.1007/s12223-012-0195-5 Authors František Golais, Faculty of Natural Sciences, Department of Microbiology and Virology, Comenius University, Mlynská dolina B2, 842 15 Bratislava, Slovakia Jaroslav Hollý, Faculty of Natural Sciences, Department of Microbiology and Virology, Comenius University, Mlynská dolina B2, 842 15 Bratislava, Slovakia Jana Vítkovská, Faculty of Natural Sciences, Department of Microbiology and Virology, Comenius University, Mlynská dolina B2, 842 15 Bratislava, Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The content of phospholipid fatty acids (PLFA) was determined in samples of polyvinyl alcohol lenses (Lentikats Biocatalyst, LB) with encapsulated Paracoccus denitrificans withdrawn during long-term denitrification experiments. The total PLFA content correlated highly with specific denitrification activities of LB as well as biomass estimation based on image analyses of microscopic photos. The results confirmed the applicability of PLFA determination for estimation of the amount of living encapsulated microbial biomass during biotechnological applications. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0189-3 Authors Josef Trögl, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Ivana Jirková, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Petra Zemánková, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Věra Pilařová, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Petra Dáňová, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Jana Pavlorková, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Pavel Kuráň, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Jan Popelka, Faculty of the Environment, Jan Evangelista Purkyně University, Králova Výšina 3132/7, 400 96 Ústí nad Labem, Czech Republic Lucie Křiklavová, Faculty of Mechatronics, Informatics and Interdisciplinary Studies, Technical University of Liberec, Studentská 2, 461 17 Liberec, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Green tea polyphenols (GTP) are widely believed to function as antioxidants and antimicrobial agents. Here we observed that GTP and epigallocatechin gallate, the most abundant catechin in GTP, could also function as prooxidants and produce hydrogen peroxide (H 2 O 2 ) to inhibit the growth of Pseudomonas aeruginosa . pH value of the medium was the key factor that affected prooxidant versus antioxidant property of GTP. Under weakly acidic conditions (pH 5.5–6.5), GTP showed antioxidant activity by eliminating H 2 O 2 ; whereas, under neutral and weakly alkaline conditions (pH 7.0–8.0), GTP showed prooxidant activity and inhibited the growth of P. aeruginosa . Furthermore, we studied the effects of GTP on gene expression profiles of a few oxidative stress-related genes by quantitative real-time PCR analysis. After 10 min to 1 h of exposure under weakly alkaline condition, GTP significantly up-regulated expression levels of katB , sodM , ohr , lexA , and recN gene. These findings highlight that the pH-dependent H 2 O 2 production by GTP contributes to the antibacterial activity and can induce oxidative stress-related responses in P. aeruginosa . Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0198-2 Authors Xiaoxiang Liu, Faculty of Basic Medicine, Zhejiang Medical College, Hangzhou, Zhejiang 310053, China Jianrong Li, Food Safety Key Lab of Liaoning Province, Bohai University, Jinzhou, Liaoning 121013, China Yanbo Wang, College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, Zhejiang 310035, China Tingting Li, College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, Zhejiang 310035, China Jin Zhao, College of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, Zhejiang 310035, China Chaohua Zhang, Huangdong Ocean University, Zhanjiang, Guangdong 524088, China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Inflammatory processes play an important role in the development of nasal polyps (NP), but the etiology and, to a high degree also, the pathogenesis of NP are not fully understood. The role of several cytokines and chemokines such as eotaxins, IL-4, IL-5, IL-6, IL-8, and RANTES has been reported in NP. Herewith, we investigated the expression and pattern of distribution of chemokine receptors CCR1 and CCR3 in nasal polyps. Immunohistochemical detection was carried out in frozen sections of biopsies from 22 NP and 18 nasal mucosa specimens in both the epithelial and stromal compartments. Fluorescence microscopy and computerized image analysis revealed a statistically significant increased number of CCR1 (45.2 ± 2.8 vs. 15.1 ± 1.9, p  〈 0.001)-positive as well as CCR3 (16.4 ± 1.4 vs. 9.7 ± 1.1, p  〈 0.001)-positive cells in the stroma of NP compared to nasal mucosa. In comparison to healthy nasal mucosa, increased positivity of CCR3 was detected in the epithelial compartment of NP. Our data suggest that increased expression of CCR1 and CCR3 chemokine receptors may, in accord with various chemokines, contribute to the pathogenesis of nasal polyposis by facilitating increased migration and prolonged accumulation of inflammatory cells, e.g., eosinophils, in the inflammatory infiltrate of NP. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0194-6 Authors P. Fundová, ENT Department of the 3rd Faculty of Medicine, Charles University and the Central Military Hospital, U vojenské nemocnice 1200, 169 02 Prague 6, Czech Republic D. P. Funda, Department of Immunology and Gnotobiology, Institute of Microbiology, v.v.i, Academy of Sciences of the Czech Republic, Prague, Czech Republic D. Kovář, ENT Department of the 3rd Faculty of Medicine, Charles University and the Central Military Hospital, U vojenské nemocnice 1200, 169 02 Prague 6, Czech Republic R. Holý, ENT Department of the 3rd Faculty of Medicine, Charles University and the Central Military Hospital, U vojenské nemocnice 1200, 169 02 Prague 6, Czech Republic M. Navara, ENT Department of the 3rd Faculty of Medicine, Charles University and the Central Military Hospital, U vojenské nemocnice 1200, 169 02 Prague 6, Czech Republic H. Tlaskalová-Hogenová, Department of Immunology and Gnotobiology, Institute of Microbiology, v.v.i, Academy of Sciences of the Czech Republic, Prague, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Twelve white-rot fungal strains belonging to seven different species were screened on plates under alkaline condition to study the decolourisation of the textile dyes Reactive Black 5 and Poly R-478. Three strains of Trametes versicolor (Micoteca da Universidade do Minho (MUM) 94.04, 04.100 and 04.101) and one strain of Phanerochaete chrysosporium (MUM 94.15) showed better decolourisation results. These four strains were used for decolourisation studies in liquid culture medium. All four selected strains presented more efficient decolourisation rates on Reactive Black 5 than on Poly R-478. For both dyes on solid and liquid culture media, the decolourisation capability exhibited by these strains depended on dye concentration and pH values of the media. Finally, the decolourisation of Reactive Black 5 by T. versicolor strains MUM 94.04 and 04.100 reached 100 %. In addition, the highest white-rot fungi ligninolytic enzyme activities were found for these two strains. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0196-4 Authors Cristiane A. Ottoni, IBB—Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal Cledir Santos, IBB—Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal Zofia Kozakiewicz, IBB—Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal Nelson Lima, IBB—Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Lactobacillus pobuzihii is a novel species which has been previously found in pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan. However, the lactic acid bacteria (LAB) microflora in pobuzihi has not been studied in detail. In this study, LAB from pobuzihi were isolated, identified, and characterized. A total of 196 LAB were isolated; 79 cultures were isolated from the sample collected from a manufacturing factory, 38 from pobuzihi samples collected from 4 different markets, and 79 from 2 fresh cummingcordia samples. These isolates were characterized phenotypically and then divided into eight groups (A to H) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Lactobacillus plantarum was the most abundant LAB found in most samples during the fermentation of pobuzihi . On the other hand, Enterococcus casseliflavus and Weissella cibaria were, respectively, the major species found in the two fresh cummingcordia samples. A potential novel species or subspecies of lactococcal strain was found. In addition, seven L. plantarum and five W. cibaria strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157 T . This is the first report describing the distribution and varieties of LAB existing in the pobuzihi during its fermentation process and the final product on the market. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0188-4 Authors Yi-sheng Chen, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan, Taoyuan, 333 Taiwan Hui-chung Wu, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan, Taoyuan, 333 Taiwan Chiung-mei Wang, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan, Taoyuan, 333 Taiwan Chia-chun Lin, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan, Taoyuan, 333 Taiwan Yi-ting Chen, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan, Taoyuan, 333 Taiwan Yu-jyun Jhong, Department of Biotechnology, Ming Chuan University, No. 5 De-Ming Road, Gui-Shan, Taoyuan, 333 Taiwan Fujitoshi Yanagida, The Institute of Enology and Viticulture, Yamanashi University, 1-13-1 Kitashin, Kofu, Yamanashi, 400-0005 Japan Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Cellulase (CEL) presently constitutes a major group of industrial enzyme based on its diverse ranges of utilization. Apart from such current and well-established applications—as in cotton processing, paper recycling, detergent formulation, juice extraction, and animal feed additives—their uses in agricultural biotechnology and bioenergy have been exploited. Supplementation of CELs to accelerate decomposition of plant residues in soil results in improved soil fertility. So far, applying CELs/antagonistic cellulolytic fungi to crops has shown to promote plant growth performance, including enhanced seed germination and protective effects. Their actions are believed mainly to trigger plant defense mechanisms and/or to act as biocontrol agents that mediate disease suppression. However, the exact interaction between the enzymes/fungi and plants has not been clearly elucidated. Under mild conditions, removal of plant cell wall polysaccharides by CELs for protoplast preparation results in reduced protoplast damage and increased viability and yields. CELs have recently shown great potential in enzyme aid extraction of bioactive compounds from plant materials before selective extraction through enhancing release of target molecules, especially those associated with the wall matrix. To date, attempts have been made to formulate CEL preparation for cellulosic-based bioethanol production. The high cost of CELs has created a bottleneck, resulting in an uneconomic production process. The utilization of low-cost carbohydrates, strain improvement, and gene manipulations has been alternatively aimed at reducing the cost of CEL production. In this review, we focus on and discuss current knowledge of CELs and their applications in agriculture, biotechnology, and bioenergy. Content Type Journal Article Pages 1-14 DOI 10.1007/s12223-012-0184-8 Authors Paripok Phitsuwan, Enzyme Technology Laboratory, Division of Biochemical Technology, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, Bangkuntien, Bangkok, 10150 Thailand Natta Laohakunjit, Flavor Technology Laboratory, Division of Biochemical Technology, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, Bangkuntien, Bangkok, 10150 Thailand Orapin Kerdchoechuen, Flavor Technology Laboratory, Division of Biochemical Technology, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, Bangkuntien, Bangkok, 10150 Thailand Khin Lay Kyu, Enzyme Technology Laboratory, Division of Biochemical Technology, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, Bangkuntien, Bangkok, 10150 Thailand Khanok Ratanakhanokchai, Enzyme Technology Laboratory, Division of Biochemical Technology, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, Bangkuntien, Bangkok, 10150 Thailand Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    A rifampicin-resistant variant of two strains of Lactobacillus plantarum , one strain of Pediococcus acidilactici , and one strain of Enterococcus faecium were used for the experimental production of lucerne silage. Laboratory silage without inoculants served as a control. Counts of total anaerobes, total lactic acid bacteria (LAB), lactobacilli, pediococci, and enterococci were determined on days 14, 21, 30, 49, and 60 of lucerne fermentation. LAB dominated in silage microflora, reaching a percentage between 59 and 95 % of total anaerobes. Lactobacilli were found as a predominant group of LAB during the whole study. Lactobacilli reached numbers 8.74 log CFU/g in treated silage and 8.89 log CFU/g in the control at the first observation. Their counts decreased to 4.23 and 4.92 log CFU/g in treated silage and the control, respectively, on day 63 of fermentation. Similar decreases were observed in all bacterial groups. The treated silage samples possessed lower pH (4.2 vs. 4.5 in control samples) and contained more lactic acid compared to control silage. The identity of re-isolated rifampicin-resistant bacteria with those inoculated to the lucerne was evaluated by fingerprinting techniques. The fingerprint profiles of re-isolated bacteria corresponded to the profiles of strains used for the treatment. It could be concluded that supplemented LAB dominated in laboratory silage and overgrew naturally occurring LAB. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0142-5 Authors Eva Vlková, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, Prague 6, 165 21 Czech Republic Vojtěch Rada, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, Prague 6, 165 21 Czech Republic Věra Bunešová, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, Prague 6, 165 21 Czech Republic Šárka Ročková, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, Prague 6, 165 21 Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The use of municipal solid waste as feedstock for biogas production offers an interesting possibility for waste treatment with the beneficial effect of gaining a green energy source. The involved processes are very complex, and many different organisms connected via a dynamic food web are associated with them. These complex interactions within these microbial communities are still not clearly understood. Therefore, a phospholipid fatty acid (PLFA) profile analysis method, well established in aerobic but still not as common in anaerobic systems, was used to throw some light on this matter. In the present investigation, a 750 m³ biogas reactor (Roppen, Austria) was monitored over a half-year period. During this period, four different phases in terms of gas production could be determined: low (I), increasing (II), high (III), and decreasing (IV) gas production. In combination with the PLFA profiles, we were able to identify changes in the microbial community associated with these phases. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0136-3 Authors Thomas Schwarzenauer, Department of Microbiology, University of Innsbruck, Technikerstraße 25d, 6020 Innsbruck, Austria Paul Illmer, Department of Microbiology, University of Innsbruck, Technikerstraße 25d, 6020 Innsbruck, Austria Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    In this work, capability of Fusarium solani F-552 of producing lignocellulose-degrading enzymes in submerged fermentation was investigated. The enzyme cocktail includes hydrolases (cellulases, xylanases, and proteinases) as well as ligninolytic enzymes: manganese-dependent peroxidase (MnP), lignin peroxidase (LiP), and laccase (Lac). To our knowledge, this is the first report on production of MnP, LiP, and Lac together by one F. solani strain. The enzyme productions were significantly influenced by application of either lignocellulosic material or chemical inducers into the fermentation medium. Among them, corn bran significantly enhanced especially productions of cellulases and xylanases (248 and 170 U/mL, respectively) as compared to control culture (11.7 and 29.2 U/mL, respectively). High MnP activity (9.43 U/mL, control 0.45 U/mL) was observed when (+)-catechin was applied into the medium, the yield of LiP was maximal (33.06 U/mL, control 2.69 U/mL) in gallic acid, and Lac was efficiently induced by, 2,2′-azino-bis-[3-ethyltiazoline-6-sulfonate] (6.74 U/mL, not detected in control). Finally, in order to maximize the ligninolytic enzymes yields, a novel strategy of introduction of mild oxidative stress conditions caused by hydrogen peroxide into the fermentation broth was tested. Hydrogen peroxide significantly increased activities of MnP, LiP, and Lac which may indicate that these enzymes could be partially involved in stress response against H 2 O 2 . The concentration of H 2 O 2 and the time of the stress application were optimized; hence, when 10 mmol/L H 2 O 2 was applied at the second and sixth day of cultivation, the MnP, LiP, and Lac yields reached 21.67, 77.42, and 12.04 U/mL, respectively. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0098-5 Authors Stanislav Obruca, Centre for Materials Research, Faculty of Chemistry, Brno University of Technology, Purkynova 118, 612 00 Brno, Czech Republic Ivana Marova, Centre for Materials Research, Faculty of Chemistry, Brno University of Technology, Purkynova 118, 612 00 Brno, Czech Republic Petra Matouskova, Centre for Materials Research, Faculty of Chemistry, Brno University of Technology, Purkynova 118, 612 00 Brno, Czech Republic Andrea Haronikova, Centre for Materials Research, Faculty of Chemistry, Brno University of Technology, Purkynova 118, 612 00 Brno, Czech Republic Andrea Lichnova, Centre for Materials Research, Faculty of Chemistry, Brno University of Technology, Purkynova 118, 612 00 Brno, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Twenty-four acid- and bile-tolerant lactobacilli isolates from dairy products were identified and further in vitro characterized for the presence of functional traits potentially useful for probiotic applications, which included desirable and undesirable traits, such as biofilm formation, ability to inhibit intestinal pathogens, antibiotic susceptibility, and enzyme activity. The majority of examined strains were susceptible to certain antimicrobial agents (streptomycin, gentamicin, clindamycin, erythromycin, tetracycline, quinupristin–dalfopristin), except for three strains of Lactobacillus rhamnosus with minimal inhibitory concentration levels for streptomycin higher than the microbiological breakpoints (≥32 μg/mL), which are considered as resistant. Undesirable traits such as α-chymotrypsin or N -acetyl-β-glucosaminidase activities were not detected, but low β-glucuronidase, and moderate and high β-glucosidase activities were recorded in nine strains, which were eliminated from further examination together with three isolates showing unsuitable antibiotic resistance. Of the remaining 12 isolates, 4 ( Lactobacillus fermentum 202, Lactobacillus gallinarum 7001, L. rhamnosus 183, and Lactobacillus plantarum L2-1) manifested an outstanding potential to inhibit selected intestinal pathogens in an agar spot test, including Escherichia coli and Salmonella spp., and simultaneously demonstrated strong biofilm-forming capacity. In conclusion, the results of our in vitro experiments showed that the above four strains had a potential probiotic value and met the criteria to be identified as a possible probiotic microorganism, with the necessity of verification through well-designed in vivo experimental, clinical, and technological studies before the strains can be used as probiotics or as starter probiotic cultures. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0121-x Authors Dobroslava Bujňáková, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, Košice, 04001 Slovakia Vladimír Kmeť, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, Košice, 04001 Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Human milk (HM) contains as the third most abundant component around 200 different structures of human milk oligosaccharides (HMOs). HMOs are the first and irreplaceable prebiotics for infants, supporting bifidobacteria as the most important bacterial group in an infant intestine. The aim of our study was to test the growth of bifidobacteria in HM and on HMOs. Bifidobacteria were isolated from two groups of infants. The first one (eight strains) were isolated from infants who had bifidobacteria in their feces but, after a short period of time (4 to 24 days), bifidobacteria were no longer detected in their feces (disappeared bifidobacteria [DB]). The second group of bifidobacteria (eight strains) originated from infants with continual presence of bifidobacteria in their feces (persistent bifidobacteria [PB]). There were significant differences ( p  〈 0.05) between DB and PB groups in the ability of the strains to grow in HM. PB grew in HM, reaching counts higher than 7 log CFU/ml. In contrast, counts of DB decreased from 5 to 4.3 log CFU/ml after cultivation in HM. The final pH after cultivation of bifidobacteria on HMOs was 6.2 and 4.9 in DP and PB groups, respectively. In general, Bifidobacterium bifidum and B. breve species were able to utilize HMOs, while B. adolescentis and B. longum subsp. longum species did not. The ability to grow in HM and to utilize HMOs seem to be important properties of bifidobacteria which are able to colonize infant intestinal tract. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0134-5 Authors Sarka Rockova, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology Food and Natural Resources, Czech University of Life Sciences, Kamycka 129, Prague 6, 165 21 Czech Republic Vojtech Rada, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology Food and Natural Resources, Czech University of Life Sciences, Kamycka 129, Prague 6, 165 21 Czech Republic Jiri Nevoral, Department of Paediatrics, 2nd Faculty of Medicine, Charles University in Prague, V Uvalu 84, Prague, 5 150 06 Czech Republic Petr Marsik, Joint Laboratory of Institute of Experimental Botany Academy of Sciences of the Czech Republic and Research Institute of Crop Production, Rozvojova 263, Prague, 6 165 02 Czech Republic Eva Vlkova, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology Food and Natural Resources, Czech University of Life Sciences, Kamycka 129, Prague 6, 165 21 Czech Republic Vera Bunesova, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology Food and Natural Resources, Czech University of Life Sciences, Kamycka 129, Prague 6, 165 21 Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Strains identified in ovine cheese and bryndza by matrix-assisted laser desorption/ionization time-of-flight analysis belonged to ten species of non-enterococcal lactic acid bacteria and included Lactobacillus casei / Lactobacillus paracasei , Lactobacillus plantarum , Lactobacillus rhamnosus , Lactobacillus helveticus , Lactobacillus delbrueckii , Lactobacillus fermentum , Lactobacillus brevis , Lactococcus lactis , Pediococcus pentosaceus and Pediococcus acidilactici . The susceptibility toward antibiotics was determined in lactobacilli, lactococci and pediococci and also in Escherichia coli for comparison. Analysis of L. fermentum and pediococci revealed the presence of non-wild-type epidemiological cut-offs in streptomycin, clindamycin or gentamicin. E. coli were resistant to ampicillin, tetracycline, enrofloxacin and florfenicol. No extended spectrum β-lactamases were detected. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0128-3 Authors V. Kmeť, Institute of Animal Physiology, Slovak Academy of Sciences, 040 01 Košice, Slovakia Z. Drugdová, Institute of Animal Physiology, Slovak Academy of Sciences, 040 01 Košice, Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The qualitative and quantitative changes in the bacterial community composition in two mesophilic, commercially used biogas plants were monitored by denaturing gradient gel electrophoresis (DGGE) and real-time PCR. The main objective was to evaluate the influence of the co-substrate maize silage on total bacteria and some selected bacterial groups by comparing full-scale reactors fed solely with pig manure or additionally with maize silage. DGGE fingerprints reflected shifts in the bacterial community structure associated with maize silage as co-substrate and the real-time PCR results showed clear changes in the quantitative composition of the bacterial consortia of each fermenter. A clear dominance of Clostridia in all surveyed fermenters and considerably lower abundance of Bacteroidetes in the biogas plant fed with maize silage was shown. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0125-6 Authors K. Fliegerová, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague, Czech Republic J. Mrázek, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague, Czech Republic M. Kajan, ENKI, o.p.s., Dukelská 1, Třeboň, Czech Republic S. M. Podmirseg, Institute of Microbiology, University of Innsbruck, Technikerstr. 25d, 6020 Innsbruck, Austria H. Insam, Institute of Microbiology, University of Innsbruck, Technikerstr. 25d, 6020 Innsbruck, Austria Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    16S rRNA sequences of ruminal uncultured bacterial clones from public databases were phylogenetically examined. The sequences were found to form two unique clusters not affiliated with any known bacterial species: cluster of unidentified sequences of free floating rumen fluid uncultured bacteria (FUB) and cluster of unidentified sequences of bacteria associated with rumen epithelium (AUB). A set of PCR primers targeting 16S rRNA of ruminal free uncultured bacteria and rumen epithelium adhering uncultured bacteria was designed based on these sequences. FUB primers were used for relative quantification of uncultured bacteria in ovine rumen samples. The effort to increase the population size of FUB group has been successful in sulfate reducing broth and culture media supplied with cellulose. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0135-4 Authors Jaime A. Rosero, Institute of Animal Physiology and Genetics, Vídeňská 1083, 142 20 Prague 4, Czech Republic Lenka Štrosová, Institute of Animal Physiology and Genetics, Vídeňská 1083, 142 20 Prague 4, Czech Republic Jakub Mrázek, Institute of Animal Physiology and Genetics, Vídeňská 1083, 142 20 Prague 4, Czech Republic Kateřina Fliegerová, Institute of Animal Physiology and Genetics, Vídeňská 1083, 142 20 Prague 4, Czech Republic Jan Kopečný, Institute of Animal Physiology and Genetics, Vídeňská 1083, 142 20 Prague 4, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Antibacterial effect of chitooligosaccharides (COS) and low molar mass chitosans (LMWC) is considered as one of the most important characteristics of chitosan (CS) hydrolysates. Here, we show the in vitro effect of different COS, LMWC, and CS on representative anaerobic bacteria isolated from human colon as a possibility of targeting modification of colonic microflora composition by supplementation of dietary CS products by humans. Specific growth rate of seven selected nonpathogenic anaerobic bacterial strains ( Clostridium paraputrificum , Clostridium beijerinckii , Roseburia intestinalis , Bacteroides vulgatus , Bacteriodes thetaiotaomicron , Faecalibacterium prausnitzii and Blautia coccoides ) was determined in the presence of 0.25 and 0.5% COS (2, 3, and 6 kDa), 0.025 and 0.05% of LMWC (10 and 16 kDa), and 0.025 and 0.1% of CS in vitro. The growth rate decreased in all strains in the presence of COS and LMWC in higher concentrations in comparison to control incubations. A relatively higher resistance to CS hydrolyzates was detected in R. intestinalis and F. prausnitzii , and more susceptible were bacteria belonging to Bacteoides sp. and Clostridium sp. The antimicrobial activity, minimum inhibitory concentrations (MIC), and minimal bactericidal concentrations (MBC) were determined. The antimicrobial activity increased with the degree of polymerization (DP). MIC ranged from 0.25 to 4.5% in dependence on bacterial strain and DP of CS/LMWC. MBC also decreased with DP. The most effective antimicrobial action was detected in LMWC with 16 kDa and CS. Weak antimicrobial activity was found in COS with small molecules (2 and 3 kDa). Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0138-1 Authors Jiří Šimůnek, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague 4, Czech Republic Věra Brandysová, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague 4, Czech Republic Ingrid Koppová, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague 4, Czech Republic Jiří Šimůnek, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague 4, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The ability of the rumen ciliates to utilize β-glucans other than cellulose and xylan is currently being recognized. The objective of the present study was to characterize the ability of the ciliate Diploplastron affine to digest some pachyman, laminarin, pustulan, curdlan and lichean. The protozoa were isolated from the rumen of sheep and either grown in vitro or inoculated into the rumen of ciliate-free sheep and maintained in natural conditions. In vitro culture studies showed that the enrichment of culture medium with the examined saccharides results in an increase in the number of ciliates in comparison to the control cultures. The increase was over 36 and 15 % when the growth medium was supplemented with pachyman (1,3-β-glucan) and pustulan (1,6-β-glucan), respectively. A positive correlation was also found between the population density of ciliates and the dose of saccharide supplemented to the growth medium. Enzyme studies were performed using the crude enzyme preparation obtained from ciliates treated with antibiotics. The ability of ciliates to digest the examined β-glucans was tested by the quantification of reducing sugars released from the mentioned substrates during the incubation with crude enzyme preparation. The results showed that D. affine ciliates were able to digest both of them. The mean degradation rate varied between 6.7 and 28.2 μmol/L glucose per mg protein per h for pustulan and lichean, respectively, whereas the digestion velocity was the highest at 5.0–5.5 pH and 45–50°C. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0120-y Authors Grzegorz Belzecki, The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences, 05-110 Jabłonna, Poland Renata Miltko, The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences, 05-110 Jabłonna, Poland Tadeusz Michalowski, The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences, 05-110 Jabłonna, Poland Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    This study was aimed to examine the genetic characteristics of 44 Streptococcus suis type 2 (SS2) isolates and the virulence attributes of 23 representative isolates. Multilocus sequence typing revealed five sequence types (ST1, ST7, ST28, ST86, and ST162) with 19 isolates assigned to ST7 (43.2%), 14 to ST1 (31.8%), and 9 to ST28 (20.5%). PCR typing of the pilus gene clusters classified the isolates into three types: A (72.7%), B (22.7%), and N (4.5%). All isolates of pilus types A and N were assigned to the ST1 complex containing ST1, ST7, and ST86, while those of type B belonged to the ST27 complex comprising ST28 and ST162. Only two strains had the putative pathogenicity island 89-kb cluster (89K) and were of type N. The type B strains had a significantly lower adhesion, were more readily killed by macrophage, and had lower virulence to mice than those of types A and N. We conclude that SS2 strains of both ST1 and ST27 complexes, parallel to pilus types A and B, were prevalent in the pig populations in Zhejiang Province, and ST7 and ST1 strains were the predominant genotypes in the diseased pigs with pneumonia. Content Type Journal Article Pages 541-548 DOI 10.1007/s12223-011-0077-2 Authors Yulong Tang, Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029 China Huancan Zhao, Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029 China Wei Wu, Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029 China Di Wu, Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029 China Xiaoliang Li, Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029 China Weihuan Fang, Institute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029 China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632 Journal Volume Volume 56 Journal Issue Volume 56, Number 6

Description:    In the present study, genetic diversity and mycotoxin profiles of Aspergillus flavus isolated from air (indoors and outdoors), levels (surfaces), and soils of five hospitals in Southwest Iran were examined. From a total of 146 Aspergillus colonies, 63 isolates were finally identified as A. flavus by a combination of colony morphology, microscopic criteria, and mycotoxin profiles. No Aspergillus parasiticus was isolated from examined samples. Chromatographic analyses of A. flavus isolates cultured on yeast extract–sucrose broth by tip culture method showed that approximately 10% and 45% of the isolates were able to produce aflatoxin B 1 (AFB 1 ) and cyclopiazonic acid (CPA), respectively. Around 40% of the isolates produced sclerotia on Czapek–Dox agar. The isolates were classified into four chemotypes based on the ability to produce AF and CPA that majority of them (55.5%) belonged to chemotype IV comprising non-mycotoxigenic isolates. Random amplified polymorphic DNA (RAPD) profiles generated by a combination of four selected primers were used to assess genetic relatedness of 16 selected toxigenic and non-toxigenic isolates. The resulting dendrogram demonstrated the formation of two separate clusters for the A. flavus comprised both mycotoxigenic and non-toxigenic isolates in a random distribution. The obtained results in this study showed that RAPD profiling is a promising and efficient tool to determine intra-specific genetic variation among A. flavus populations from hospital environments. A. flavus isolates, either toxigenic or non-toxigenic, should be considered as potential threats for hospitalized patients due to their obvious role in the etiology of nosocomial aspergillosis. Content Type Journal Article Pages 527-534 DOI 10.1007/s12223-011-0078-1 Authors Asghar Sepahvand, Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-331 Iran Masoomeh Shams-Ghahfarokhi, Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-331 Iran Abdolamir Allameh, Department of Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-331 Iran Zahra Jahanshiri, Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, 14115-331 Iran Mojdeh Jamali, Department of Mycology, Pasteur Institute of Iran, Tehran, 13164 Iran Mehdi Razzaghi-Abyaneh, Department of Mycology, Pasteur Institute of Iran, Tehran, 13164 Iran Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632 Journal Volume Volume 56 Journal Issue Volume 56, Number 6

Description:    Moonlighting proteins have two different functions within a single polypeptide chain. Exploring moonlighting enzymes from the environment using the metagenomic approach is interesting. In the present study, a novel β-glucosidase gene, designated as bgl1D , with lipolytic activity (renamed Lip1C) was cloned through function-based screening of a metagenomic library from uncultured soil microorganisms. The deduced amino acid sequence comparison and phylogenetic analysis also indicated that Lip1C and other putative lipases are closely related. Biochemical characterization demonstrated that the maximum activity of the recombinant Lip1C protein occurs at pH 8.0 and 30°C using 4-nitrophenyl butyrate as substrate. The putative lipase had an apparent K m value of 0.88 mmol/L, a k cat value of 212/min, and a k cat / K m value of 241 L/mmol/min. Lip1C exhibited habitat-specific characteristics with 5 mmol/L AlCl 3 , CuCl 2 , and LiCl. The characterization of the biochemical properties of Lip1C enhances our understanding of this novel moonlighting enzyme isolated from a soil metagenome. Content Type Journal Article Pages 563-570 DOI 10.1007/s12223-011-0083-4 Authors Cheng-Jian Jiang, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, and College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi 530004, People’s Republic of China Gao Chen, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, and College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi 530004, People’s Republic of China Jie Huang, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, and College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi 530004, People’s Republic of China Qin Huang, College of Chemistry and Ecology Engineering, Guangxi University for Nationalities, 188 Daxue East Road, Nanning, Guangxi 530006, People’s Republic of China Ke Jin, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, and College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi 530004, People’s Republic of China Pei-Hong Shen, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, and College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi 530004, People’s Republic of China Jun-Fang Li, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, and College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi 530004, People’s Republic of China Bo Wu, College of Chemistry and Ecology Engineering, Guangxi University for Nationalities, 188 Daxue East Road, Nanning, Guangxi 530006, People’s Republic of China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632 Journal Volume Volume 56 Journal Issue Volume 56, Number 6

Description:    There is no doubt about the beneficial effect of breastfeeding on the newborn's immune system. It is not fully elucidated what the differences are between the colostrum/milk of healthy and allergic mothers and how beneficial breastfeeding by an allergic mother is. The gene expression of selected cytokines was tested in cells isolated from colostra of healthy and allergic mothers using quantitative real-time PCR. Allergic phenotype was evident in colostral cells of allergic mothers: gene expressions of IL-4, IL-13 and EGF were increased and those of IFN-gamma decreased in comparison with colostral cells of healthy mothers. The allergic phenotype of the colostral cells of allergic mothers supporting the bias to a Th2 type response was found. It remains a question if a small number of these cells could influence the immature newborn immune system. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0112-y Authors Jiří Hrdý, Institute of Immunology and Microbiology, First Faculty of Medicine, Charles University in Prague, Studnickova 7, 128 00 Prague 2, Czech Republic Olga Novotná, Institute of Immunology and Microbiology, First Faculty of Medicine, Charles University in Prague, Studnickova 7, 128 00 Prague 2, Czech Republic Ingrid Kocourková, Institute for the Care of Mother and Child, Prague, Czech Republic Ludmila Prokešová, Institute of Immunology and Microbiology, First Faculty of Medicine, Charles University in Prague, Studnickova 7, 128 00 Prague 2, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    One of the main mechanisms of nanoparticle toxicity is known to be the generation of reactive oxygen species (ROS) which primarily damage cell membranes. However, very limited data on membrane effects in anaerobic environments (where ROS could not be the cause of membrane damage) are available. In the following study, rumen anaerobe Ruminococcus flavefaciens 007C was used as a bacterial model to assess the potential effects of Al 2 O 3 and TiO 2 nanoparticles on membranes in an anaerobic environment. Fatty acid profiles of cultures after exposure to Al 2 O 3 or TiO 2 nanoparticles were analyzed and compared with the profiles of non-exposed cultures or cultures exposed to bulk materials. Analysis revealed dose–effect changes in membrane composition exclusively when cells were exposed to Al 2 O 3 nanoparticles in a concentration range of 3–5 g/L, but were not present in cultures exposed to bulk material. On the other hand, the tested concentrations of nano-TiO 2 did not significantly affect the membrane profile of the exposed bacterium. The results suggest the possibility that Al 2 O 3 induces changes in bacterial membranes by direct physical interaction, which was supported by TEM image analysis. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0143-4 Authors Maša Vodovnik, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Rok Kostanjšek, Department of Biology, University of Ljubljana, Večna pot 111, 1000 Ljubljana, Slovenia Maša Zorec, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Romana Marinšek Logar, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The expression of Ruminococcus flavefaciens 007S cellulases in different incubation time points (growth stages) and their substrate inducibility were analyzed by comparing the zymogram expression profiles of cultures grown on insoluble cellulose (Avicel) with cellobiose-grown cultures. The molecular weights of the enzymes were compared to (putative) cellulases encoded in the R. flavefaciens FD-1 genome. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0144-3 Authors Maša Vodovnik, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Romana Marinšek Logar, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    We endeavored to develop a method for viability determination of solventogenic clostridia and to apply it for monitoring acetone–butanol–ethanol (ABE) fermentation. Six fluorescent probes (propidium iodide [PI], ethidium bromide, fluorescein diacetate, carboxyfluorescein diacetate [cFDA], rhodamine 123, bis-(1,3-dibutylbarbituric acid)trimethine oxonol [BOX]) were tested in order to distinguish two subpopulations of live and dead clostridial cells in suspension. Three of them were found to be appropriate (PI, BOX and cFDA) for this purpose. Developed fluorescent staining methods were applied to batch fermentation processes of Clostridium pasteurianum and C. beijerinckii carried out in a laboratory bioreactor under anaerobic conditions. Whereas PI was found to be applicable to both strains, BOX was convenient only for viability determination of C. pasteurianum . Although cFDA can distinguish two cell subpopulations in suspension, it was found to be unsuitable for viability determination under tested conditions, since it reflected more variable esterase activity during sporulation cell cycle than viability. Flow cytometry in combination with convenient fluorescent probe has been proved to be a valuable tool for viability determination. We assume this rapid and simple method can help to obtain more complex and precise information about ABE fermentation. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0131-8 Authors Michaela Linhová, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 166 28 Prague, Czech Republic Barbora Branská, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 166 28 Prague, Czech Republic Petra Patáková, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 166 28 Prague, Czech Republic Jakub Lipovský, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 166 28 Prague, Czech Republic Petr Fribert, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 166 28 Prague, Czech Republic Mojmír Rychtera, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 166 28 Prague, Czech Republic Karel Melzoch, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 166 28 Prague, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Fecal bacteria from 33 infants (aged 1 to 6 months) were tested for growth on commercial prebiotics. The children were born vaginally (20) or by caesarean section (13). Bifidobacteria, lactobacilli, gram-negative bacteria, Escherichia coli , and total anaerobes in fecal samples were enumerated by selective agars and fluorescence in situ hybridization. The total fecal bacteria were inoculated into cultivation media containing 2 % Vivinal® (galactooligosaccharides—GOS) or Raftilose® P95 (fructooligosaccharides—FOS) as a single carbon source and bacteria were enumerated again after 24 h of anaerobic cultivation. Bifidobacteria dominated, reaching counts of 9–10 log colony-forming units (CFU)/g in 17 children born vaginally and in seven children delivered by caesarean section. In these infants, lactobacilli were more frequently detected and a lower number of E. coli and gram-negative bacteria were determined compared to bifidobacteria-negative infants. Clostridia dominated in children without bifidobacteria, reaching counts from 7 to 9 log CFU/g. Both prebiotics supported all groups of bacteria tested. In children with naturally high counts of bifidobacteria, bifidobacteria dominated also after cultivation on prebiotics, reaching counts from 8.23 to 8.77 log CFU/mL. In bifidobacteria-negative samples, clostridia were supported by prebiotics, reaching counts from 7.17 to 7.69 log CFU/mL. There were no significant differences between bacterial growth on Vivinal® and Raftilose® P95 and counts determined by cultivation and FISH. Prebiotics should selectively stimulate the growth of desirable bacteria such as bifidobacteria and lactobacilli. However, our results showed that commercially available FOS and GOS may stimulate also other fecal bacteria. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0123-8 Authors Věra Bunešová, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, 165 21 Prague 6, Czech Republic Eva Vlková, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, 165 21 Prague 6, Czech Republic Vojtěch Rada, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, 165 21 Prague 6, Czech Republic Vladimíra Kňazovická, Department of Microbiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, 949 76 Nitra, Slovakia Šárka Ročková, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, 165 21 Prague 6, Czech Republic Martina Geigerová, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, 165 21 Prague 6, Czech Republic Matěj Božik, Department of Microbiology, Nutrition and Dietetics, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, 165 21 Prague 6, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Genetic diversity of the isolates of Fusarium oxysporum f. sp. ciceris causing chickpea wilt collected from 12 states representing different agro-ecological regions of India was determined through randomly amplified polymorphic DNA (RAPD) markers. The UPGMA cluster analysis grouped the isolates into eight categories showing high magnitude of genetic diversity. Each group had the isolates from different states present in various agro-ecological regions of India. Therefore, the groups generated through the RAPD analysis were not corresponding to area of the origin of the isolates. The RAPD primers, namely, OPA 7 and OPA 11 produced Foc specific fragment of ≈1.3 kb and ≈1.4 kb, respectively in all the isolates. These fragments were eluted, purified, cloned in pGEM-T Easy vector and sequenced. Primers were designed with sequence information of these two fragments using primer.3 software. Two sets of sequence characterized amplified region markers (SC-FOC 1 and SC-FOC 2) developed from the sequences of these fragments were found to be specific to Foc and produced an amplicon of 1.3 and 1.4 kb, respectively. These set of markers were validated against the isolates of the pathogen collected from different locations of India representing various races of the pathogen. They are non-specific to the other Fusarium species, Rhizoctonia solani and R. bataticola. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0118-5 Authors M. Durai, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110 012 India Sunil C. Dubey, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110 012 India Aradhika Tripathi, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110 012 India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The ability of rumen ciliates to digest chitin is clearly recognized. We investigated the chitinolytic system of the rumen ciliate Eudiplodinium maggii . The ciliates were grown in a selectively faunated sheep. They were isolated from the rumen and purified by sedimentation. A crude enzyme preparation was prepared following incubation of ciliates with antibiotics. This was done in order to reduce their contamination with intracellular bacteria. The activity of particular enzymes was examined by quantification of the products released from specific substrates. It was stated that the optimum conditions for the detected activities varied between 4.5 and 5.5 pH, and 45 and 55 °C. β- N -Acetylglucosaminidase was found as an enzyme of the highest activity (4.2 μmol/l released product per mg protein per h). The activities of endochitinase and exochitinase were almost two times lower than that of β- N -acetylglucosaminidase. Zymographic studies revealed the presence of two endochitinases, two exochitinases and two β- N -acetylglucosaminidases in the examined preparation. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0133-6 Authors R. Miltko, The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences, Instytucka 3, 05-110 Jabłonna near Warsaw, Poland G. Belzecki, The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences, Instytucka 3, 05-110 Jabłonna near Warsaw, Poland T. Michalowski, The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences, Instytucka 3, 05-110 Jabłonna near Warsaw, Poland Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Recently the ATP-binding cassette (ABC) efflux pumps have been proved to be a major component of drug resistance in Mycobacterium tuberculosis . The objective of this study was to investigate the expression profiles of Rv1456c-Rv1457c-Rv1458c efflux system in clinical isolates of M. tuberculosis and its involvement in drug-resistance mechanisms. Significantly increased mRNA expression of Rv1456c, Rv1457c, and Rv1458c appeared among the clinical isolates ( P  〈 0.05), which are resistant to at least one of the four first-line drugs including rifampin, isoniazid, streptomycin, and ethambutol. In addition, overexpression of this efflux system was more frequently found in multidrug-resistant and extensively drug-resistant M. tuberculosis strains. Therefore, Rv1456c-Rv1457c-Rv1458c efflux pumps may play an important role in drug resistance of treatment of M. tuberculosis . Further investigation of this gene may lead to the development of countermeasures against M. tuberculosis drug resistance. Content Type Journal Article Pages 549-553 DOI 10.1007/s12223-011-0080-7 Authors Pei Hao, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Zhang Shi-Liang, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Liu Ju, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Dai Ya-Xin, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Huang Biao, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Wang Xu, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Hu Min-Tao, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Kuai Shou-Gang, Wuxi Hospital of Infectious Disease, Wuxi, 214005 Jiangsu Province, China Wang Ke, Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063 Jiangsu Province, China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632 Journal Volume Volume 56 Journal Issue Volume 56, Number 6

Description:    Soil microbial populations play crucial role in soil properties and influence below-ground ecosystem processes. Microbial composition and functioning changes the soil quality through decomposition of organic matter, recycling of nutrients, and biological control of parasites of plants. Moreover, the discovery that soil microbes may translate into benefits for biotechnology, management of agricultural, forest, and natural ecosystems, biodegradation of pollutants, and waste treatment systems maximized the need of scientists for the isolation and their characterization. Operations such as the production of antibiotics and enzymic activities from microorganisms of soil constitute objectives of industry in her effort to cope with the increase of population of earth and disturbance of environment and may ameliorate the effects of global climate change. In the past decades, new biochemical and molecular techniques have been developed in our effort to identify and classify soil bacteria. The goal of measuring the soil microbial diversity is difficult because of the limited knowledge about bacteria species and classification through families and orders. Molecular techniques extend our knowledge about microbial diversity and help the taxonomy of species. Measuring and monitoring soil microbial communities can lead us to better understanding of their composition and function in many ecosystem processes. Content Type Journal Article Pages 1-8 DOI 10.1007/s12223-012-0179-5 Authors Christos Stefanis, Faculty of Agricultural Development, Department of Food Science and Technology, Laboratory of Microbiology, Biotechnology and Hygiene, Democritus University of Thrace, 193 Pandazidou Str, 68200 Orestiada, Greece Athanasios Alexopoulos, Faculty of Agricultural Development, Department of Food Science and Technology, Laboratory of Microbiology, Biotechnology and Hygiene, Democritus University of Thrace, 193 Pandazidou Str, 68200 Orestiada, Greece Chrissa Voidarou, Faculty of Agricultural Development, Department of Food Science and Technology, Laboratory of Microbiology, Biotechnology and Hygiene, Democritus University of Thrace, 193 Pandazidou Str, 68200 Orestiada, Greece Stavros Vavias, Faculty of Agricultural Development, Department of Food Science and Technology, Laboratory of Microbiology, Biotechnology and Hygiene, Democritus University of Thrace, 193 Pandazidou Str, 68200 Orestiada, Greece Eugenia Bezirtzoglou, Faculty of Agricultural Development, Department of Food Science and Technology, Laboratory of Microbiology, Biotechnology and Hygiene, Democritus University of Thrace, 193 Pandazidou Str, 68200 Orestiada, Greece Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description: Prevalence of chromosomally integrated HHV-6 in patients with malignant disease and healthy donors in the Czech Republic Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0180-z Authors Petr Hubacek, Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine of Charles University and Motol University Hospital, V Uvalu 84, Prague 5, CZ 150 06 Czech Republic Alena Hrdlickova, Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine of Charles University and Motol University Hospital, V Uvalu 84, Prague 5, CZ 150 06 Czech Republic Martin Spacek, Department of Clinical Haematology, 3rd Faculty of Medicine of Charles University and University Hospital Kralovske Vinohrady, Prague, Czech Republic Miroslav Zajac, Department of Medical Microbiology, Motol University Hospital, Prague, Czech Republic Katerina Muzikova, Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine of Charles University and Motol University Hospital, V Uvalu 84, Prague 5, CZ 150 06 Czech Republic Petr Sedlacek, Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine of Charles University and Motol University Hospital, V Uvalu 84, Prague 5, CZ 150 06 Czech Republic Petr Cetkovsky, Institute of Haematology and Blood Transfusion, U nemocnice 1, Prague, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Candidemia is a major infectious complication in neonatal patients. The isolation of yeasts from blood is still the “gold standard” for its diagnosis, but other laboratory markers (i.e., circulating antigens) have been studied with varying specificities and sensitivities. The aim of this study was to evaluate the role of procalcitonin for the diagnosis of candidemia in neonatal patients at high risk. To verify if the use of different commercial methods can highlight dissimilar results of sensitivity and/or specificity, the determination of procalcitonin serum levels was estimated by two systems. Overall, 90 patients from a Neonatal Intensive Care Units were enrolled, of whom six developed Candida bloodstream infection. Four of six infants with candidemia had slight increase of procalcitonin values (0.5–1 ng/mL). Only one baby showed very high levels but he had fungal and bacterial sepsis at the same time, while no elevation was observed in the sixth patient. No statistically significant difference was observed between two different methods at the time of monitoring ( p  〉 0.643). Both methods showed a sensitivity of 83.3 % at diagnosis, while the specificity was 73.8 and 63.1 % by methods A and B, respectively. In the light of the low sensibility and specificity of this assay, we can assume that the determination of procalcitonin would not seem to play a significant role in the diagnosis of fungal infection in neonatal patients. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0169-7 Authors Maria Teresa Montagna, Department of Biomedical Science and Human Oncology, University of Bari “Aldo Moro”, Piazza G. Cesare 11, 70124 Bari, Italy Caterina Coretti, Department of Biomedical Science and Human Oncology, University of Bari “Aldo Moro”, Piazza G. Cesare 11, 70124 Bari, Italy Antonella Rella, Department of Biomedical Science and Human Oncology, University of Bari “Aldo Moro”, Piazza G. Cesare 11, 70124 Bari, Italy Giovanna Barbuti, Department of Biomedical Science and Human Oncology, University of Bari “Aldo Moro”, Piazza G. Cesare 11, 70124 Bari, Italy Fabio Manca, Department of Historical and Geographic Science, Statistic Unit, University of Bari “Aldo Moro”, Via Quintino Sella, 268-70123 Bari, Italy Osvaldo Montagna, Department of Gynecology, Obstetrics and Neonatology, Neonatology and Neonatology Intensive Care Section, University of Bari “Aldo Moro”, Piazza G. Cesare 11, 70124 Bari, Italy Nicola Laforgia, Department of Gynecology, Obstetrics and Neonatology, Neonatology and Neonatology Intensive Care Section, University of Bari “Aldo Moro”, Piazza G. Cesare 11, 70124 Bari, Italy Giuseppina Caggiano, Department of Biomedical Science and Human Oncology, University of Bari “Aldo Moro”, Piazza G. Cesare 11, 70124 Bari, Italy Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The kinetic study of Arthrospira platensis extracellular polymeric substances (EPS) production under different trophic modes—photoautotrophy (100 μmol photons m −2  s −1 ), heterotrophy (1.5 g/L glucose), and mixotrophy (100 μmol photons m −2  s −1 and 1.5 g/L glucose)—was investigated. Under photoautotrophic and heterotrophic conditions, the maximum EPS production 219.61 ± 4.73 and 30.30 ± 1.97 mg/L, respectively, occurred during the stationary phase. Under a mixotrophic condition, the maximum EPS production (290.50 ± 2.21 mg/L) was observed during the early stationary phase. The highest specific EPS productivity (433.62 mg/g per day) was obtained under a photoautotrophic culture. The lowest specific EPS productivity (38.33 mg/g per day) was observed for the heterotrophic culture. The effects of glucose concentration, light intensity, and their interaction in mixotrophic culture on A. platensis EPS production were evaluated by means of 32 factorial design and response surface methodology. This design was carried out with a glucose concentration of 0.5, 1.5, and 2.5 g/L and at light levels of 50, 100, and 150 μmol photons m −2  s −1 . Statistical analysis of the model demonstrated that EPS concentration and EPS yield were mainly influenced by glucose concentration and that conditions optimizing EPS concentration were dissimilar from those optimizing EPS yield. The highest maximum predicted EPS concentration (369.3 mg/L) was found at 150 μmol photons m −2  s −1 light intensity and 2.4 g/L glucose concentration, while the highest maximum predicted EPS yield (364.3 mg/g) was recorded at 115 μmol photons m −2  s −1 light intensity and 1.8 g/L glucose concentration. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0170-1 Authors Lamia Trabelsi, Laboratory of Marine Biodiversity and Biotechnology, National Institute of Marine Sciences and Technology, BP 59, 5000 Monastir, Tunisia Hatem Ben Ouada, Laboratory of Marine Biodiversity and Biotechnology, National Institute of Marine Sciences and Technology, BP 59, 5000 Monastir, Tunisia Fatma Zili, Laboratory of Marine Biodiversity and Biotechnology, National Institute of Marine Sciences and Technology, BP 59, 5000 Monastir, Tunisia Nahla Mazhoud, Laboratory of Marine Biodiversity and Biotechnology, National Institute of Marine Sciences and Technology, BP 59, 5000 Monastir, Tunisia Jihen Ammar, Laboratory of Marine Biodiversity and Biotechnology, National Institute of Marine Sciences and Technology, BP 59, 5000 Monastir, Tunisia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Five new strains of lactobacilli isolated from goatling’s stomach were identified by molecular–biological approaches. Profiles of fermentable saccharides, Gram staining, and cell morphology were also determined. They were identified as Lactobacillus reuteri (strains KO4b, KO4m, KO5) and as Lactobacillus plantarum (strains KG1z, KG4). In DNA samples of all newly isolated L. reuteri strains as well as in L. reuteri E (Lreu E; originated from lamb), the part of gld C gene, coding large subunit of glycerol dehydratase, that is necessary for 3-hydroxypropionaldehyde (3-HPA; reuterin) production, was amplified using two designed primer sets. However, the 3-HPA production was revealed only in the strain Lreu E. It produced five- or ten-fold lower amount of 3-HPA in comparison with probiotic L. reuteri ATCC 55730 in aerobic or anaerobic conditions, respectively. Moreover, Lreu E completely lost its production ability after ca. five passages in MRS medium. The co-incubation of Lreu E, but not other L. reuteri isolates, with Escherichia coli re-induced 3-HPA production. In the case of L. reuteri ATCC 55730, the 3-HPA production increased more than four times after co-incubation with E. coli . Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0166-x Authors Hana Kiňová Sepová, Department of Cellular and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University in Bratislava, Bratislava, Slovakia Andrea Bilková, Department of Cellular and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University in Bratislava, Bratislava, Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Helicobacter pylori was examined in 110 patients (82 (74.5) with gastritis, 18 (16.4) with duodenitis, six (5.5) with duodenal ulcer and gastroesophageal reflux, and four (3.6 %) with normal) with gastrointestinal problems living in rural area, no history of macrolide use, and detected by culture (71.8) or direct detection from gastric biopsies by PCR (82.7 %). Also, cagA gene was identified using PCR and was found positive in 68/91 (74.7 %) strains. The prevalence of clarithromycin-resistant H. pylori was investigated by two methods including PCR–RFLP (7.7 (A2142G 1.1 and A2143G 6.6 %)) and twofold agar dilution (8.9 %) to detect phenotypic and genotypic status simultaneously. Among all the H. pylori positive patients, eight (8.8 %) isolates were found to be resistant to clarithromycin by at least one of the AD and/or PCR–RFLP methods . H. pylori positive rates were significantly correlated with patients' sex, age, and endoscopic findings ( p  = 0.040, 〈0.001 and 〈0.001, respectively). There were no differences in gender or endoscopic findings related to cagA + and cagA − patients. The gene of cagA was not significantly helpful in predicting the clinical outcome of H. pylori infection alone. In conclusion, we revealed that there was a low prevalence of primer clarithromycin resistance in patients living in rural area with no history of macrolide use. The prevalence of mutant strains among the macrolide-resistant H. pylori varies even geographically between close provinces. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0192-8 Authors Erkan Yula, Department of Medical Microbiology, Faculty of Medicine, University of Mustafa Kemal, Antakya, Hatay 31100, Turkey Toğrul Nagiyev, Department of Medical Microbiology, Faculty of Medicine, University of Cukurova, Adana, Turkey Özlem Aycan Kaya, Department of Medical Parasitology, Faculty of Medicine, University of Mustafa Kemal, Hatay, Turkey Melek İnci, Department of Medical Microbiology, Faculty of Medicine, University of Mustafa Kemal, Antakya, Hatay 31100, Turkey M. Murat Çelik, Department of Internal Medicine, Faculty of Medicine, University of Mustafa Kemal, Hatay, Turkey Fatih Köksal, Department of Medical Microbiology, Faculty of Medicine, University of Cukurova, Adana, Turkey Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Ginkgo biloba has long been used in traditional Chinese medicine. In this study, ginkgoneolic acid, a kind of compound extracted from G. biloba , was investigated for its effects on growth, acid production, adherence, biofilm formation, and biofilm morphology of Streptococcus mutans . The results showed that ginkgoneolic acid inhibited not only the growth of S. mutans planktonic cells at minimum inhibitory concentration (MIC) of 4 μg/mL and minimum bactericidal concentration (MBC) of 8 μg/mL but also the acid production and adherence to saliva-coated hydroxyapatite of S. mutans at sub-MIC concentration. In addition, this agent was effective in inhibiting the biofilm formation of S. mutans (MBIC 50  = 4 μg/mL), and it reduced 1-day-developed biofilm of S. mutans by 50 % or more at low concentration (MBRC 50  = 32 μg/mL) . Furthermore, the present study demonstrated that ginkgoneolic acid disrupted biofilm integrity effectively. These findings suggest that ginkgoneolic acid is a natural anticariogenic agent in that it exhibits antimicrobial activity against S. mutans and suppresses the specific virulence factors associated with its cariogenicity. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0191-9 Authors Jinzhi He, State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China Shida Wang, State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China Tingxi Wu, State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China Yangpei Cao, State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China Xin Xu, State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China Xuedong Zhou, State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Four local Bacillus thuringiensis (Bt) isolates that had been serologically identified as Bt var . kurstaki (Btk2, Btk3, and Btk66) and Bt var. mexicanensis (Btm27), in addition to two reference strains (4D20 and 4AC1), were laboratory assayed as microbial control agents against the Egyptian cotton leafworm Spodoptera littoralis (Boisd.). Polymerase chain reaction (PCR) amplification analysis revealed that each of the six experimental strains carries, at least, a cry1 type gene which expresses a protein toxin active against lepidopterous insects. Additionally, PCR amplification results demonstrated that 4D20 and Btk66 contain the Lepidoptera- and Diptera-active cry2 type gene and that Btk66 contains Coleoptera-active cry7 and cry8 genes. Among the six strains, Btk66 and Btm27 were the most promising microbial control agents against S. littoralis . The present findings were the first to report that Btm27 (classified as B. thuringiensis var. mexicanensis ) is a very potent microbial control agent against S. littoralis -tested larvae. For more characterization of these two isolates, the sspO gene was investigated as a molecular chronometer. The DNA sequencing results proved that Btk66 and Btm27 carry sspO open reading frames with identical nucleotide sequences, suggesting a strong phylogenetic relationship between the two strains. Content Type Journal Article Pages 1-8 DOI 10.1007/s12223-012-0193-7 Authors Ahlam A. Alfazairy, Faculty of Agriculture, Alexandria University, Alexandria, Egypt Amani M. D. El-Ahwany, Botany and Microbiology Department, Faculty of Science, Alexandria University, Alexandria, Egypt Eman A. Mohamed, Faculty of Science, Damanhour University, Damanhour, Egypt Heba A. H. Zaghloul, Botany and Microbiology Department, Faculty of Science, Alexandria University, Alexandria, Egypt Ehab R. El-Helow, Botany and Microbiology Department, Faculty of Science, Alexandria University, Alexandria, Egypt Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Plasmalogens are a group of lipids with potentially important, and not yet fully known, functions in organisms from bacteria to protozoans, invertebrates, and mammals. They can protect cells against the damaging effects of reactive oxygen species, protect other phospholipids or lipoprotein particles against oxidative stress, and have been implicated as signaling molecules and modulators of membrane dynamics. They have been found in many anaerobic bacterial species, and their biosynthetic pathways differ in aerobic and anaerobic organisms. The use of advanced techniques permits the identification of not only plasmalogen classes but also their positional isomers and often also individual molecular species. This paper describes direct analyses of plasmalogens from natural sources, frequently very unusual, using electrospray ionization mass spectrometry in combination with high-performance liquid chromatography and/or shotgun lipidomics. Content Type Journal Article Pages 1-10 DOI 10.1007/s12223-012-0178-6 Authors Tomáš Řezanka, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Prague 4, Czech Republic Zdena Křesinová, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Prague 4, Czech Republic Irena Kolouchová, Department of Biotechnology, Institute of Chemical Technology Prague, Technická 5, 16628 Prague, Czech Republic Karel Sigler, Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Prague 4, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description: Infection by Neisseria meningitidis serogroup W135 belonging to the unusual clone ST-3342 in the Czech Republic Content Type Journal Article Pages 1-2 DOI 10.1007/s12223-012-0177-7 Authors Helena Janouškovcová, Department of Microbiology, Faculty of Medicine and University Hospital, Charles University, Plzen, Czech Republic Jaroslav Raděj, 1st Medical Department, ICU, Faculty of Medicine and University Hospital, Charles University, Plzen, Czech Republic Martin Musílek, National Reference Laboratory for Meningococcal Infections, National Institute of Public Health, Prague, Czech Republic Zuzana Vacková, National Reference Laboratory for Meningococcal Infections, National Institute of Public Health, Prague, Czech Republic Jana Kozáková, National Reference Laboratory for Meningococcal Infections, National Institute of Public Health, Prague, Czech Republic Paula Kriz, National Reference Laboratory for Meningococcal Infections, National Institute of Public Health, Prague, Czech Republic Tamara Bergerová, Department of Microbiology, Faculty of Medicine and University Hospital, Charles University, Plzen, Czech Republic Jaroslav Hrabák, Department of Microbiology, Faculty of Medicine and University Hospital, Charles University, Plzen, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description: 7th International Symposium on Anaerobic Microbiology, ISAM 2011 Content Type Journal Article Pages 1-1 DOI 10.1007/s12223-012-0153-2 Authors Peter Javorský, Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia Peter Pristaš, Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia Jiri Simunek, Institute of Animal Physiology and Genetics v.v.i, Academy of Sciences of the Czech Republic, Prague, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The genus Vibrio is characterized by a large number of species and some of them are human pathogens causing gastrointestinal and wound infections through the ingestion or manipulation of contaminated fishes and shellfish including Vibrio parahaemolyticus and Vibrio alginolyticus . In this study, we reported the phenotypic and molecular characterization of 9 V. parahaemolyticus and 27 V. alginolyticus strains isolated from outbreaks affecting cultured Gilthead sea bream ( Sparus aurata L.) and Sea bass ( Dicentrarchus labrax ) along the Tunisian coast from 2008 to 2009. All isolates were tested for the presence of DNase, caseinase, protease, lipase, amylase, gelatinase, hemolytic activity and antibacterial resistance to different drugs. Randomly amplified polymorphic DNA was used to examine the genetic relatedness among the V. parahaemolyticus and V. alginolyticus strains. Content Type Journal Article Pages 1-10 DOI 10.1007/s12223-012-0174-x Authors Khouadja Sadok, Laboratoire d’Analyse, Traitement et Valorisation des Polluants de l’Environnement et des Produits, Département de Microbiologie, Faculté de Pharmacie, Rue Avicenne, 5000 Monastir, Tunisia Snoussi Mejdi, Laboratoire de Traitement des Eaux Usées. Centre de Recherches et des Technologies des Eaux, Technopole de Borj-Cédria, BP 901, 2050 Hammam-Lif, Tunisia Saidi Nourhen, Laboratoire d’Analyse, Traitement et Valorisation des Polluants de l’Environnement et des Produits, Département de Microbiologie, Faculté de Pharmacie, Rue Avicenne, 5000 Monastir, Tunisia Bakhrouf Amina, Laboratoire d’Analyse, Traitement et Valorisation des Polluants de l’Environnement et des Produits, Département de Microbiologie, Faculté de Pharmacie, Rue Avicenne, 5000 Monastir, Tunisia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The aim of present work was to study chemical structures and biological activities of rhamnolipid biosurfactants produced by Pseudomonas aeruginosa MN1 isolated from oil-contaminated soil. The results of liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed that total rhamnolipids (RLs) contained 16 rhamnolipid homologues. Di-lipid RLs containing C 10 -C 10 moieties were by far the most predominant congeners among mono-rhamnose (53.29 %) and di-rhamnose (23.52 %) homologues. Mono-rhamnolipids form 68.35 % of the total congeners in the RLs. Two major fractions were revealed in the thin layer chromatogram of produced RLs which were then purified by column chromatography. The retardation factors ( R f ) of the two rhamnolipid purple spots were 0.71 for RL1 and 0.46 for RL2. LC-MS/MS analysis proved that RL1 was composed of mono-RLs and RL2 consisted of di-RLs. RL1 was more surface-active with the critical micelle concentration (CMC) value of 15 mg/L and the surface tension of 25 mN/m at CMC. The results of biological assay showed that RL1 is a more potent antibacterial agent than RL2. All methicillin-resistant Staphylococcus aureus (MRSA) strains were inhibited by RLs that were independent of their antibiotic susceptibility patterns. RLs remarkably enhanced the activity of oxacillin against MRSA strains and lowered the minimum inhibitory concentrations of oxacillin to the range of 3.12–6.25 μg/mL. Content Type Journal Article Pages 1-8 DOI 10.1007/s12223-012-0164-z Authors Nasrin Samadi, Department of Drug and Food Control and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 14171 Iran Neda Abadian, Department of Drug and Food Control and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 14171 Iran Reza Ahmadkhaniha, Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences, Tehran, 14171 Iran Farzaneh Amini, Department of Drug and Food Control and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 14171 Iran Dina Dalili, Department of Drug and Food Control and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 14171 Iran Noushin Rastkari, Institute for Environmental Research, Tehran University of Medical Sciences, Tehran, Iran Eliyeh Safaripour, Pharmaceutical Quality Assurance Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 14171 Iran Farzaneh Aziz Mohseni, Biotechnology Department, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    A phosphate solubilizing fungus, Aspergillus awamori S29 was isolated from rhizoshpere of mungbean. The phosphate solubilizing activity of A. awamori S29 in liquid was 1,110 mg/L for tricalcium phosphate (TCP). The organism was able to solubilize various inorganic forms of phosphate at a wide range of temperatures. Among various insoluble phosphate sources tested, di-calcium phosphate was solubilized the most, followed by TCP. A. awamori S29 had significant effect ( p  〈 0.05) on mungbean growth, total P and plant biomass under pot conditions, although no obvious difference in available P in soil and number of leaves was found compared to the control. Content Type Journal Article Pages 1-9 DOI 10.1007/s12223-012-0167-9 Authors Rachana Jain, Department of Bioscience and Biotechnology, Banasthali University, Banasthali, 304022 Rajasthan, India Jyoti Saxena, Biochemical Engineering Department, BT Kumaon Institute of Technology, Dwarahat, 263653 Uttarakhand, India Vinay Sharma, Department of Bioscience and Biotechnology, Banasthali University, Banasthali, 304022 Rajasthan, India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Pseudomonads have been reported for their metabolic, nutritional and ecological versatility, which motivated us to prospect the metabolic profile of a lipolytic strain Pseudomonas aeruginosa SL-72. The strain SL-72 was found to produce high levels of lipase and pectinase (1,555.62 IU/mL and 1,490.33 IU/mL, respectively), esterase and amylase, besides low levels of xylanase, proteinase and cellulase. The strain also tested positive for different plant growth-promoting traits—production of ammonia, hydrogen cyanide, siderophores, phosphate solubilization, nitrate reduction and antifungal activity. The high levels of activity of aryl sulphatase, alkaline phosphatase and fluorescein diacetate hydrolase makes it a useful strain for enhanced nutrient cycling in soil. The strain SL-72 produced rhamnolipids, a biosurfactant and its production was enhanced when starch was used as carbon source (0.256 g/L) and utilized polycyclic hydrocarbon compounds viz. anthracene, phenanthrene, pyrene, fluorene and its mixture. The multifaceted nature of the culture illustrates its promise in bioremediation, industry, besides its use as an inoculant. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0163-0 Authors Shikha Verma, Department of Biosciences and Biotechnology, Banasthali University, Banasthali, 304022 India Radha Prasanna, Division of Microbiology, Indian Agricultural Research Institute, New Delhi, 110012 India Jyoti Saxena, Biochemical Engineering Department, BCT Kumaon Engineering College, Dwarahat, 263653 Uttarakhand, India Vinay Sharma, Department of Biosciences and Biotechnology, Banasthali University, Banasthali, 304022 India Lata Nain, Division of Microbiology, Indian Agricultural Research Institute, New Delhi, 110012 India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Small plasmid pKST23 was isolated from sheep ruminal Escherichia coli population. Plasmid sequence was determined to be 2,779 bp in length and was found to have an overall 42 % of GC pairs. However, its sequence can be divided into two regions based on genetic composition and the GC content. It was found that the high GC region spanning approximately from nucleotide 1,300 to 2,750 was identical to a group of small Escherichia coli plasmids and encoded a putative replication protein identical to plasmid pKL1 Rep protein. The part with lower GC pairs seemed to be more specific as it showed no similarity to the GenBank database. Computational analysis revealed four open reading frames, two of which showed considerable homology to replication proteins. PCR primers targeting parts of the two different regions of plasmid pKST23 were used to assess the occurrence of related plasmids within ruminal E. coli population. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0124-7 Authors L. Fecskeová, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovak Republic M. Kovařík, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovak Republic P. Javorský, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovak Republic P. Pristaš, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovak Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The ibeA gene, one of the important invasion-associated genes in neonatal meningitis Escherichia coli (NMEC), has been recently detected in avian pathogenic E. coli (APEC). Thus, it is necessary to close monitor the possible contamination of the poultry farms and its products to people. Here, a dot blot method for detecting the ibeA gene in E. coli was developed and validated. For the present study, probe sequence was designed and optimized for the specificity of dot blot. A 342-bp conserved fragment of ibeA gene was selected and labeled with digoxigenin (DIG)-dUTP according to the manufacturer’s guidelines, which indicated that this probe hybridizes with ibeA . In our established method, the bacteria culture samples were directly spotted on the membrane, following simple lyses on the membrane. Hence, the extraction of genomic DNA is not required, which reduces the workload and shortens the time. Furthermore, this assay was very sensitive, which could detect as few as 2.5 × 10 3  CFU bacteria. The diagnostic reliability of this dot blot was evaluated on 467 APEC bacteria samples by using PCR analysis. Both methods showed that the result was in complete concordance. The dot blot assay was proved to be a simple, rapid, highly accurate, and cost-effective method to identify invasion-associated genes ibeA , which could be applied for initial screening of a large number of clinical samples or direct detection of bacteria culture. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0171-0 Authors Chunling Niu, Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, 210095 China Shaohui Wang, Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, 210095 China Chengping Lu, Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, 210095 China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Melanins are complex natural pigments that darken the skin and are difficult to degrade. This study evaluated synthetic melanin decolorization by the crude laccase from fungus Lentinus polychrous in the absence and presence of selected redox mediators. The greatest melanin decolorization activity was 87 % at pH 6.5 within 3 h in the presence of 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonate) diammonium salt (ABTS), whereas only about 22 % melanin decolorized at pH 5.0 in case of no mediator. The optimum temperatures for melanin decolorization in the absence and presence of ABTS were 55 and 35°C, respectively. Using a natural redox mediator, 1.0 mmol/L vanillin leads to 45 % melanin decolorization. Our results suggest the possibility of applying vanillin for L. polychrous laccase-catalyzed decolorization of melanin. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0151-4 Authors Saranyu Khammuang, Protein and Enzyme Technology Research Unit and Center of Excellence for Innovation in Chemistry, Department of Chemistry, Faculty of Science, Mahasarakham University, Maha Sarakham, 44150 Thailand Rakrudee Sarnthima, Protein and Enzyme Technology Research Unit and Center of Excellence for Innovation in Chemistry, Department of Chemistry, Faculty of Science, Mahasarakham University, Maha Sarakham, 44150 Thailand Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The cold shock protein family consists of the transfer of the foodborne pathogen Listeria monocytogenes from 37 to 4 and −20 °C and was characterized by the sharp induction of a low molecular mass protein. This major cold shock protein ferritin-like protein (Flp) has an important role in regulation of various microbial physiological processes. Flp have a molecular mass of about 18 kDa, as observed on SDS–PAGE. The purification procedure including ammonium sulfate fractionation was used. Monospecific polyclonal antibodies raised in rabbits against the purified new Flp immunostained a single 18-kDa Flp band in extracts from different cytoplasmic proteins blotted onto nitrocellulose. A 411-bp cDNA fragment that corresponds to an internal region of an flp gene was obtained by RT-PCR. Our result indicated a surexpression of major cold shock protein and an important increase in flp mRNA amount after a downshift temperature especially at −20 °C. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0172-z Authors Hanene Miladi, Laboratoire d’Analyses, Traitement et Valorisation des Polluants de l’Environnement et des Produits, Faculté de Pharmacie, rue Avicenne, 5000 Monastir, Tunisia Abdelaziz Soukri, Laboratoire de Physiologie et de Génétique Moléculaire (PGM), Faculté des Sciences, Ain-Chock,, BP 5366, Maarif Casablanca, Morocco Amina Bakhrouf, Laboratoire d’Analyses, Traitement et Valorisation des Polluants de l’Environnement et des Produits, Faculté de Pharmacie, rue Avicenne, 5000 Monastir, Tunisia Emna Ammar, Unité de Recherche Gestion des Environnements Urbains et Côtiers–LARSEN, Ecole Nationale d’Ingénieurs de Sfax (Tunisie), B.P. «W», 3038 Sfax, Tunisia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Four simple sets for digital microphotography are described that have been tested with the Carl Zeiss Jena, Meopta Prague, Lambda Prague, and LOMO Sankt Petersburg microscopes and with DSLR Nikon D 70 and Nikon D 300 cameras. They permit precise image focusing in the camera using a prism Zeiss. The sets make use of commonly available extensions Zeiss, Praktica and reductions Nikon—Praktica manufactured by ROWI (without a lens) or HAMA (with a lens). An extension has further been designed and manufactured for connecting the DSLR Nikon D 300 camera fitted with the HAMA reduction (only with a lens) and a focusing extensible prism with Zeiss Jena light measurement. It permits a precise image focusing of low light intensity objects (autofluorescence or low-contrast or moving objects when using positive or negative phase contrast). The sets are applicable to all microscopes constructed according to German DIN industrial standards. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0162-1 Authors Zdeněk Žižka, Laboratory of Molecular Structure Characterization, Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i., 142 20 Prague, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Genetic diversity of 89 isolates of Rhizoctonia solani isolated from different pulse crops representing 21 states from 16 agro-ecological regions of India, 49 morphological, and 7 anastomosis groups (AGs) was analyzed using 12 universal rice primers (URPs), 22 random amplified polymorphic DNA (RAPD), and 23 inter-simple sequence repeats (ISSR) markers. Both URPs and RAPD markers provided 100 % polymorphism with the bands ranging from 0.1 to 5 kb in size, whereas ISSR markers gave 99.7 % polymorphism with the bands sizes ranging from 0.1 to 3 kb. The marker URP 38F followed by URP13R, URP25F, and URP30F, RAPD marker R1 followed by OPM6, A3 and OPA12 and ISSR3 followed by ISSR1, ISSR4, and ISSR20 produced the highest number of amplicons. R. solani isolates showed a high level of genetic diversity. Unweighted pair group method with an arithmetic average (UPGMA) analysis grouped the isolates into 7 major clusters at 35 % genetic similarity using the three sets of markers evaluated. In spite of using three different types of markers, about 95 % isolates shared common grouping patterns. The majority of the isolates representing various AGs were grouped together into different sub-clusters using all three types of markers. Molecular groups of the isolates did not correspond to agro-ecological regions or states and crops of the origin. An attempt was made for the first time in the present study to determine the genetic diversity of R. solani populations isolated from different pulse crops representing various AGs and agro-ecological regions. Content Type Journal Article Pages 1-12 DOI 10.1007/s12223-012-0165-y Authors Sunil C. Dubey, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110 012 India Aradhika Tripathi, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110 012 India B. K. Upadhyay, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, 110 012 India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The antibiotic L-155,175, a potent antiparasitic and antifungal compound, has an unusual structure involving 16-membered macrolides that contain a tetrahydropyran ring connected through a three-carbon linker chain. To identify the biosynthetic gene cluster for L-155,175, a genomic DNA library of Streptomyces hygroscopicus ATCC31955 was constructed and screened with a degenerate primer set designed from a conserved region of the ketosynthase (KS) domain. Sequence analysis of a fosmid clone, pEY1D8 (34 kb), revealed multiple open reading frames (ORFs) encoding type I polyketide synthase (PKS). To determine whether the cloned genes are involved in L-155,175 biosynthesis, a deletion mutant (1D8m) was generated by homologous recombination, in which the gene encoding the KS domain was substituted with an apramycin-resistance gene by PCR-targeted Streptomyces gene replacement. LC–MS analysis showed that L-155,175 production was completely abolished in the 1D8m strain, thereby proving that the cloned gene is responsible for L-155,175 biosynthesis. The sequencing of two other fosmid clones (pEY8B10 and pEY1C9) harboring overlapping sequences from pEY1D8 revealed a 60-kb DNA segment encoding six ORFs for type I PKS harboring 12 modules. The domain organization of the PKS modules encoded by PKS exactly matched the structure of L-155,175. This is the first report on the gene cluster involved in the biosynthesis of L-155,175. Content Type Journal Article Pages 1-8 DOI 10.1007/s12223-012-0173-y Authors Eun Young Kim, Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, 5 Anam-dong, Seongbuk-gu, Seoul, 136-713 Republic of Korea Jae Woo Han, Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, 5 Anam-dong, Seongbuk-gu, Seoul, 136-713 Republic of Korea Jee Yeon Lee, Department of Chemistry, The Scripps Research Institute, 130 Scripps Way, Jupiter, FL 33458, USA Beom Seok Kim, Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, 5 Anam-dong, Seongbuk-gu, Seoul, 136-713 Republic of Korea Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Bioinformatic evidence of the presence of a large conjugative transposon in ruminal bacterium Prevotella bryantii B 1 4 T is presented. The described transposon appears to be related to another large conjugative transposon CTnBST, described in Bacteroides uniformis WH207 and to the conjugative transposon CTn3-Bf, which was observed in the genome of Bacteroides fragilis strain YCH46. All three transposons share tra gene regions with high amino acid identity and clearly conserved gene order. Additionally, a second conserved region consisting of hypothetical genes was discovered in all three transposons and named the GG region. This region served as a specific sequence signature and made possible the discovery of several other apparently related hypothetical conjugative transposons in bacteria from the genus Bacteroides . A cluster of genes involved in sugar utilization and metabolism was discovered within the hypothetical CTnB 1 4, to a certain extent resembling the polysaccharide utilization loci which were described recently in some Bacteroides strains. This is the first firm report on the presence of a large mobile genetic element in any strain from the genus Prevotella . Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0126-5 Authors Katja Gorenc, Biotechnical Faculty, Animal Science Department, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Tomaž Accetto, Biotechnical Faculty, Animal Science Department, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Gorazd Avguštin, Biotechnical Faculty, Animal Science Department, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The ciliate Diploplastron affine is known as a common species of the rumen fauna in cattle and sheep. This protozoon is able to digest cellulose, whereas its amylolytic activity is not well known. The objective of the reported studies was to examine the ability of D. affine to digest starch and to use this polysaccharide to cover the requirement for energy. The enzymatic studies showed that the protozoal cell extract degraded starch to reducing products with the rate being equivalent to 2.4 ± 0.47 μmol/L glucose per mg protein per min. Maltose, maltotriose and a small quantity of glucose were the end products of starch degradation. The degradation rate of maltose was only 0.05 μmol/L glucose per mg protein per min. Two peaks in α-amylase and a single peak in maltase activity were found following molecular filtration of ciliate cell extract, whereas three starch-degrading enzymes were identified by a zymographic technique. Incubation of the bacteria-free ciliates with starch in the presence of antibiotics resulted in a release of volatile fatty acids with the net rate of 25 pmol per protozoan per h. Acetic acid followed by butyric acid was the main product of starch fermentation. The results confirmed the ability of D. affine to utilize starch in energy-yielding processes. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0146-1 Authors K. Wereszka, Department of Ruminant Physiology of Nutrition, The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Instytucka 3, 05-110 Jablonna near Warsaw, Poland T. Michałowski, Department of Ruminant Physiology of Nutrition, The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Instytucka 3, 05-110 Jablonna near Warsaw, Poland Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    A strictly anaerobic mesophilic chitinolytic bacterial strain identified as Clostridium paraputrificum J4 was isolated from human feces. In response to various types of growth substrates, the bacterium produced an array of chitinolytic enzymes representing significant components of the J4 strain secretome. The excreted active proteins were characterized by estimating the enzymatic activities of endochitinase, exochitinase, and N- acetylglucosaminidase induced by cultivation in medium M-10 with colloidal chitin. The enzyme activities produced by J4 strain cultivated in medium M-10 with glucose were significantly lower. The spectrum of extracellularly excreted proteins was separated by SDS-PAGE. The chitinase variability was confirmed on zymograms of renatured SDS-PAGE. The enzymes were visualized under ultraviolet light by using 4-methylumbelliferyl derivatives of N -acetyl-β- d -glucosaminide, N , N ´-diacetyl-β- d -chitobiose, or N , N ´, N ˝-triacetyl-β- d -chitotriose for β- N -acetylglucosaminidase, chitobiosidase, or endochitinase activities, respectively. Protein components of the secretome were separated by 2D-PAGE analysis. The distinct protein bands were excised, isolated, and subsequently characterized by using MALDI-TOF/TOF tandem mass spectrometry. The final identification was performed according to sequence homology by database searching. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0137-2 Authors Jiří Šimůnek, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague 4, Czech Republic Ingrid Koppová, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, v.v.i., Vídeňská 1083, 142 20 Prague 4, Czech Republic Galina Tiščenko, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, v.v.i., Heyrovský Sq. 2, 162 06 Prague 6, Czech Republic Jan Dohnálek, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, v.v.i., Heyrovský Sq. 2, 162 06 Prague 6, Czech Republic Jarmila Dušková, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, v.v.i., Heyrovský Sq. 2, 162 06 Prague 6, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    This review surveys on the influence of different environmental factors like light (intensity, quality, photoperiod), temperature, season, nutrients (inorganic, organic), biotic factors (algal extracellular products, bacterial association, animals grazing), osmotic stress, pH of the medium, wave motion and mechanical shock, pollution, and radiations (UV, X-rays, gamma radiation) on the induction (or inhibition) of algal reproduction like cell division in unicellular algae, and formation of zoospores, aplanospores, akinetes, cysts, antheridia, oogonia, zygospores, etc. Content Type Journal Article Pages 1-21 DOI 10.1007/s12223-012-0147-0 Authors S. C. Agrawal, Department of Botany, University of Allahabad, Allahabad, 211002 India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Two hundred eighty-four isolates of enterococci from feces of wild living chamois from alpine environments were tested for sensitivity to three antibiotics. Low frequency of resistance was observed in studied enterococcal populations (about 5 % for tetracycline and erythromycin and 0 % for ampicillin). In six animals, the population of enterococci lacked any detectable resistance. Our data indicated that enterococcal population in feces of the majority of studied animals did not encounter mobile genetic elements encoding antibiotic resistance probably due to spatial separation and/or due to low exposure to the antibiotics. Based on resistance profiles observed, three populations were analyzed for the presence of restriction endonucleases. The restriction enzymes from two isolates—31K and 1K—were further purified and characterized. Restriction endonuclease Efa1KI recognizes CCWGG sequence and is an isoschizomer of BstNI. Endonuclease Efc31KI, a BsmAI isoschizomer, recognizes the sequence GTCTC and it is a first restriction endonuclease identified in Enterococcus faecium . Our data indicate that restriction–modification (R–M) systems do not represent an efficient barrier for antibiotic resistance spreading; enterococcal populations colonized by antibiotics resistance genes were also colonized by the R–M systems. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0141-6 Authors A. Vandžurová, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia I. Hrašková, Faculty of Natural Sciences, Matej Bel University, Tajovského 40, 974 01 Banská Bystrica, Slovakia J. Júdová, Faculty of Natural Sciences, Matej Bel University, Tajovského 40, 974 01 Banská Bystrica, Slovakia P. Javorský, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia P. Pristaš, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Denaturant gradient gel electrophoresis (DGGE) enables insight into the diversity of the studied microbial communities on the basis of separation of PCR amplification products according to their nucleotide sequence composition. However, the success of the method is accompanied by the inherent appearance of various sequence artifacts that bias the impression of community structure by generating additional bands representing no virtual microbes. PCR-DGGE artifacts require optimization of the method when aiming at the phylogenetic identification of the selected DGGE bands. The aim of our study was to develop a procedure which will increase the reliability of the identification. Samples of rumen fluid were used for the optimization since they contain a complex microbial community that supports the generation of artifactual bands. An optimized procedure following band excision and elution of microbial DNA is proposed including nuclease treatment, selection of DNA polymerase with proofreading activity, and cloning prior to sequencing and identification analysis. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0130-9 Authors Darja Kušar, Biotechnical Faculty, Animal Science Department, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Gorazd Avguštin, Biotechnical Faculty, Animal Science Department, Chair for Microbiology and Microbial Biotechnology, University of Ljubljana, Groblje 3, 1230 Domžale, Slovenia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The effect of the administration of chitosan (CS) and chitooligosaccharides (COS) on rat fecal microbiota was analyzed in this study. The profile of total bacterial population was monitored during 3 weeks of CS or COS application using denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene amplicons. Quantitative PCR was used for monitoring possible changes in the levels of total bacteria and the levels of individual bacterial groups: Bifidobacteria , Clostridium leptum , Enterobacteriaceae, Lactobacillus–Streptococcus–Enterobacter , and Bacteroides–Prevotella . The DGGE profiles revealed a high complexity and individuality of each tested subject, and variations in the composition of band pattern were observed. CS or COS per os administration changed the profile and structure of the microbial ecosystem of the gastrointestinal tract of healthy rats. COS have, in most cases, an opposite effect compared with CS; only the Bacteroides–Prevotella bacterial group and Enterobacteriaceae were influenced in the same way. The Bifidobacteria group was not influenced by the administration CS and COS. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0129-2 Authors Ingrid Koppová, Institute of Animal Physiology and Genetics, Academy of Sciences of Czech Republic, Vídeňská 1083, Prague, 142 20 Czech Republic Martin Bureš, Institute of Animal Physiology and Genetics, Academy of Sciences of Czech Republic, Vídeňská 1083, Prague, 142 20 Czech Republic Jiří Šimůnek, Institute of Animal Physiology and Genetics, Academy of Sciences of Czech Republic, Vídeňská 1083, Prague, 142 20 Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Real-time polymerase chain reaction (real-time PCR) is a laboratory technique based on PCR. This technique is able to detect sequence-specific PCR products as they accumulate in “real time” during the PCR amplification, and also to quantify the number of substrates present in the initial PCR mixture before amplification begins. In the present study, real-time PCR assay was employed for rapid and real-time detection of Bacillus anthracis spores spiked in 0.1 g of soil and talcum powder ranging from 5 to 10 7 spores. DNA was isolated from spiked soil and talcum powder, using PBS containing 1 % Triton-X-100, followed by heat treatment. The isolated DNA was used as template for real-time PCR and PCR. Real-time PCR amplification was obtained in 60 min under the annealing condition at 60°C by employing primers targeting the pag gene of B. anthracis . In the present study, the detection limit of real-time PCR assay in soil was 10 3 spores and10 2 spores in talcum powder, respectively, whereas PCR could detect 10 4 spores in soil and 10 3 spores in talcum powder, respectively. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0117-6 Authors Neha Jain, Division of High Containment Facility, Defence Research & Development Establishment (DRDE), Jhansi Road, Gwalior, PIN-474002 MP, India S. Merwyn, Division of High Containment Facility, Defence Research & Development Establishment (DRDE), Jhansi Road, Gwalior, PIN-474002 MP, India G. P. Rai, Division of High Containment Facility, Defence Research & Development Establishment (DRDE), Jhansi Road, Gwalior, PIN-474002 MP, India G. S. Agarwal, Division of High Containment Facility, Defence Research & Development Establishment (DRDE), Jhansi Road, Gwalior, PIN-474002 MP, India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Uncontrolled microbial methane production is playing an important role in global warming. In the present study, we showed that water content and incubation temperature increase the potential for methane formation in the two alpine soils under investigation. Beside these factors, the grazing of cows and thus the amendment of methanogenic microorganisms by cattle dung is the most important factor determining the potential of methane production in those soils. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0145-2 Authors Andreas O. Wagner, Institute of Microbiology, University of Innsbruck, Technikerstr. 25, 6020 Innsbruck, Austria Katrin Hofmann, Institute of Microbiology, University of Innsbruck, Technikerstr. 25, 6020 Innsbruck, Austria Eva Prem, Institute of Microbiology, University of Innsbruck, Technikerstr. 25, 6020 Innsbruck, Austria Paul Illmer, Institute of Microbiology, University of Innsbruck, Technikerstr. 25, 6020 Innsbruck, Austria Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    A quantitative fluorescent in situ hybridization method was employed to evaluate the competitive inhibitory effect of three Lactobacillus strains ( Lactobacillus reuteri , Lactobacillus gasseri , and Lactobacillus plantarum ) against Escherichia coli internalization in a model system of HT 29 cells. Furthermore, aggregation and adhesion abilities of the Lactobacillus strains were examined. All lactobacilli were able to attach to the HT 29 cells and aggregate with pathogens; however, the adhesion and aggregation degree was strain-dependent. L. reuteri possessed a high capacity of adhesion (6.80 ± 0.63; log CFU ± SEM per well), whereas lower capacities were expressed by L. gasseri (4.52 ± 0.55) and L. plantarum (4.90 ± 0.98). Additionally, L. reuteri showed the rapid or normal ability to aggregate with selected E. coli in comparison with remaining two lactobacilli, which showed only slow or negative aggregative reaction. Internalization of E. coli into the cell lines was markedly suppressed by L. reuteri , while L. gasseri and L. plantarum caused only a minimum anti-invasion effect. The fact that L. reuteri in our experiments showed an outstanding potential for adhering to the colon epithelial cell line, compared with the rest strains, suggested that one of the possible mechanisms of preventing pathogen adhesion and invasion is simple competitions at certain receptors and capability to block receptor binding sites, or that an avid interaction between L. reuteri and the host cell might be modulating intracellular events responsible for the E. coli internalization. Moreover, L. reuteri exhibited a strong ability to aggregate with E. coli , which could be another limiting factor of pathogen invasion. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0122-9 Authors Dobroslava Bujňáková, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, Košice, 04001 Slovakia Vladimír Kmeť, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4/6, Košice, 04001 Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Strategies are sought to reduce intestinal colonisation of food-producing animals by Campylobacter jejuni , a leading bacterial cause of human foodborne illness worldwide. Presently, we tested the antimicrobial activity of hydrolysable-rich blackberry, cranberry and chestnut tannin extracts and condensed tannin-rich mimosa, quebracho and sorghum tannins (each at 100 mg/mL) against C. jejuni via disc diffusion assay in the presence of supplemental casamino acids. We found that when compared to non-tannin-treated controls, all tested tannins inhibited the growth of C. jejuni and that inhibition by the condensed tannin-rich mimosa and quebracho extracts was mitigated in nutrient-limited medium supplemented with casamino acids. When tested in broth culture, both chestnut and mimosa extracts inhibited growth of C. jejuni and this inhibition was much greater in nutrient-limited than in full-strength medium. Consistent with observations from the disc diffusion assay, the inhibitory activity of the condensed tannin-rich mimosa extracts but not the hydrolysable tannin-rich chestnut extracts was mitigated by casamino acid supplementation to the nutrient-limited medium, likely because the added amino acids saturated the binding potential of the condensed tannins. These results demonstrate the antimicrobial activity of various hydrolysable and condensed tannin-rich extracts against C. jejuni and reveal that condensed tannins may be less efficient than hydrolysable tannins in controlling C. jejuni in gut environments containing high concentrations of amino acids and soluble proteins. Content Type Journal Article Pages 1-6 DOI 10.1007/s12223-012-0119-4 Authors Robin C. Anderson, United States Department of Agriculture/Agricultural Research Service, Southern Plains Agricultural Research Center, College Station, TX 77845, USA Maša Vodovnik, Microbiology and Microbial Biotechnology, Department of Animal Science, University of Ljubljana, Domžale, Slovenia Byeng R. Min, Texas AgriLife Research, Vernon, TX 76385, USA William E. Pinchak, Texas AgriLife Research, Vernon, TX 76385, USA Nathan A. Krueger, United States Department of Agriculture/Agricultural Research Service, Southern Plains Agricultural Research Center, College Station, TX 77845, USA Roger B. Harvey, United States Department of Agriculture/Agricultural Research Service, Southern Plains Agricultural Research Center, College Station, TX 77845, USA David J. Nisbet, United States Department of Agriculture/Agricultural Research Service, Southern Plains Agricultural Research Center, College Station, TX 77845, USA Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Actinobacteria ( Actinomycetes ) are a significant and interesting group of gram-positive bacteria. They are regular, though infrequent, members of the microbial life in the rumen and represent up to 3 % of total rumen bacteria; there is considerable lack of information about ecology and biology of rumen actinobacteria. During the characterization of variability of rumen treponemas using non-cultivation approach, we also noted the variability of rumen actinobacteria. By using Treponema -specific primers a specific 16S rRNA gene library was prepared from cow and sheep rumen total DNA. About 10 % of recombinant clones contained actinobacteria-like sequences. Phylogenetic analyses of 11 clones obtained showed the high variability of actinobacteria in the ruminant digestive system. While some sequences are nearly identical to known sequences of actinobacteria, we detected completely new clusters of actinobacteria-like sequences, representing probably new, as yet undiscovered, group of rumen Actinobacteria. Further research will be necessary for understanding their nature and functions in the rumen. Content Type Journal Article Pages 1-3 DOI 10.1007/s12223-012-0140-7 Authors M. Šuľák, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia L. Sikorová, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia J. Jankuvová, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia P. Javorský, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia P. Pristaš, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    An agar-degrading bacterium, Rhodococcus sp. Q5, was isolated from printing and dyeing wastewater using a mineral salts agar plate containing agar as the sole carbon source. The bacterium grew from pH 4.0 to 9.0, from 15 to 35°C, and in NaCl concentrations of 0–5 %; optimal values were pH 6.0, 30°C, and 1 % NaCl. Maximal agarase production was observed at pH 6.0 and 30°C. The bacterium did not require NaCl for growth or agarase production. The agarase secreted by Q5 was inducible by agar and was repressed by all simple sugars tested except lactose. Strain Q5 could hydrolyze starch but not cellulose or carboxymethyl cellulose. Agarase activity could also be detected in the medium when lactose or starch was the sole source of carbon and energy. Strain Q5 could grow in nitrogen-free mineral media; an organic nitrogen source was more effective than inorganic carbon sources for growth and agarase production. Addition of more organic nitrogen (peptone) to the medium corresponded with reduced agarase activity. Content Type Journal Article Pages 1-8 DOI 10.1007/s12223-012-0150-5 Authors Zehua Feng, College of Basic Medical and Biological Science, Soochow University, Suzhou, 215123 People’s Republic of China Lin Peng, College of Basic Medical and Biological Science, Soochow University, Suzhou, 215123 People’s Republic of China Mei Chen, College of Basic Medical and Biological Science, Soochow University, Suzhou, 215123 People’s Republic of China Mengying Li, College of Basic Medical and Biological Science, Soochow University, Suzhou, 215123 People’s Republic of China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The emergence of spa types and spa –clonal complexes (CC) among clinical methicillin-resistant Staphylococcus aureus isolates collected from the University Clinical Center in Gdańsk between 2008 and 2009 were investigated. Phage typing was used as the initial screening in the study. The basic set of phages and the additional set of phages were used. Most of the isolates (56 %) belonged to the phage group III. With the additional set of phages, eight types were found, with predominant one MR8 (50 %). Sixteen distinct spa types were observed. The most frequent were t003 (22 %), t151 (16 %), and t008 (12 %). The spa types were clustered into two spa -CC and eight singletons. The predominant CC010 (50 %) consisted of six types, with the most common t003 (36.7 %) and t151(26.7 %), and in 80 % was identified as staphylococcal chromosomal casette mec (SCC mec ) type II. The second cluster has no founder (12 %) with only two spa types: t037 belonging to SCC mec type III and t029. In the most frequent singleton, spa type t008 alone was clustered in 12 % of the isolates. All singletons correspond to SCC mec type IV. The CC010 was distributed in most of the hospital wards, corresponded to Multilocus sequence typing type ST5/ST225 and was constantly present throughout the observed period. The isolates of CC010 generally belonged to the phage group III, and most of them (53.3 %) were resistant to erythromycin, clindamycin, and ciprofloxacin. The concordance between spa -clone and phage type was very high, but the same phage type MR8 was observed within different spa t ypes of the predominant clone. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0148-z Authors Katarzyna Wiśniewska, Department of Medical Microbiology, Medical University of Gdańsk, Do Studzienki 38, 80-227 Gdańsk, Poland Anna Szewczyk, Department of Clinical Microbiology, University Clinical Center in Gdańsk, Smoluchowskiego 18, 80-214 Gdańsk, Poland Lidia Piechowicz, Department of Medical Microbiology, Medical University of Gdańsk, Do Studzienki 38, 80-227 Gdańsk, Poland Marek Bronk, Department of Clinical Microbiology, University Clinical Center in Gdańsk, Smoluchowskiego 18, 80-214 Gdańsk, Poland Alfred Samet, Department of Clinical Microbiology, University Clinical Center in Gdańsk, Smoluchowskiego 18, 80-214 Gdańsk, Poland Krystyna Świeć, Department of Medical Microbiology, Medical University of Gdańsk, Do Studzienki 38, 80-227 Gdańsk, Poland Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Although invasive fungal diseases (IFDs) are relatively rare, they have become an increasingly common life-threatening complication in a variety of critically ill patients. Due to changes in treatment strategies, patterns of IFDs have changed substantially as well. Yeast infections have shifted toward a higher proportion of non- albicans Candida species, but their overall incidence has remained stable. In contrast, IFDs caused by molds, including particularly various species of Aspergillus , Fusarium , and Mucorales, have increased in number. In view of the growing incidence and the high mortality rates of IFDs, accurate diagnostic techniques permitting timely onset of adequate antifungal treatment are of paramount importance. Although conventional approaches such as microscopy, cultivation, histopathological examination, and imaging methods still represent the gold standard, the diagnosis remains difficult because of limited sensitivity and specificity. Noninvasive and culture-independent diagnostic techniques, including fungal antigen detection, and different molecular-based techniques are becoming increasingly important. Of the fungal surrogate markers such as cell wall components, galactomannan and (1,3)-β- d -glucan by commercially available diagnostic kits have become widely used, but the results are still controversial. A plethora of PCR-based diagnostic methods targeting different gene regions and exploiting a variety of amplicon detection tools have been published. Molecular assays have the capacity to overcome the limitations of other diagnostic approaches, but the current lack of methodological standardization and validation, together with not always clear interpretation of the results, has prevented broad application in the clinical setting. Content Type Journal Article Pages 1-10 DOI 10.1007/s12223-012-0152-3 Authors Lenka Bašková, Division of Molecular Microbiology, Children’s Cancer Research Institute (CCRI), Vienna, 1090 Austria Vladimír Buchta, Department of Clinical Microbiology, Faculty of Medicine and University Hospital in Hradec Kralove, Charles University in Prague, Hradec Kralove, 500 05 Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description: Use of the manganese-dependent superoxide dismutase gene sodA for rapid identification of recently described enterococcal species Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0115-8 Authors Petra Frolkova, Department of Biology and Wildlife Diseases, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Palackeho 1-3, 612 42 Brno, Czech Republic Anuradha Ghosh, Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, USA Pavel Svec, Czech Collection of Microorganisms, Department of Experimental Biology, Faculty of Science, Masaryk University, Tvrdeho 14, 602 00 Brno, Czech Republic Ludek Zurek, Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, USA Ivan Literak, Department of Biology and Wildlife Diseases, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, Palackeho 1-3, 612 42 Brno, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The potential for comparing microbial community population structures has been greatly enhanced by developments in next generation sequencing methods that can generate hundreds of thousands to millions of reads in a single run. Conversely, many microbial community comparisons have been published with no more than 1,000 sequences per sample. These studies have presented data on levels of shared operational taxonomic units (OTUs) between communities. Due to lack of coverage, that approach might compromise the conclusions about microbial diversity and the degree of difference between environments. In this study, we present data from recent studies that highlight this problem. Also, we analyzed datasets of 16 rRNA sequences with small and high sequence coverage from different environments to demonstrate that the level of sequencing effort used for analyzing microbial communities biases the results. We randomly sampled pyrosequencing-generated 16S rRNA gene libraries with increasing sequence effort. Sequences were used to calculate Good's coverage, the percentage of shared OTUs, and phylogenetic distance measures. Our data showed that simple counts of presence/absence of taxonomic unities do not reflect the real similarity in membership and structure of the bacterial communities and that community comparisons based on phylogenetic tests provide a way to test statistically significant differences between two or more environments without need an exhaustive sampling effort. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0155-0 Authors Leandro N. Lemos, Universidade Federal do Pampa, Campus São Gabriel, Av. Antônio Trilha, 1847 São Gabriel, Rio Grande do Sul, Brazil Roberta R. Fulthorpe, Department of Physical and Environmental Sciences, University of Toronto, Scarborough, Ontario, Canada Luiz F. W. Roesch, Universidade Federal do Pampa, Campus São Gabriel, Av. Antônio Trilha, 1847 São Gabriel, Rio Grande do Sul, Brazil Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The impact of probiotic supplementation of canine-derived strain Lactobacillus fermentum AD1-CCM7421 in freeze-dried form on quantitative composition of microbiota and short-chain fatty acid profile in feces of dogs was demonstrated by two independent studies (straightforward repeated-measures model; study I: a dose of 2 g per dog for 2 weeks, 10 8 CFU/g, n  = 12; study II: 1 g per dog for 1 week, 10 7 CFU/g, n  = 11. The results revealed a significant increase of lactic acid bacteria population persisting also after the cessation of probiotic application in both studies. A reduction of clostridia (study I, p sum  〈 0.01) and tested Gram-negative bacterial genera (coliforms, Aeromonas sp., Pseudomonas sp., study II, p 〈  0.05) was also detected. The strain AD1-CCM7421 colonized the canine digestive tract in sufficient numbers (10 5 –10 6 CFU/g) and it persisted in the majority of dogs after cessation of probiotic application. An increase of short-chain fatty acid concentrations (study I: butyric, succinic, valeric, formic acid) especially in the early post-treatment phase ( p 〈  0.05) most likely led to a decrease of fecal pH value ( p 〈  0.05) without negative influence on fecal consistency throughout the studies. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0139-0 Authors V. Strompfová, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia A. Lauková, Institute of Animal Physiology, Slovak Academy of Sciences, Šoltésovej 4-6, 040 01 Košice, Slovakia S. Gancarčíková, Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy, Komenského 73, 041 81 Košice, Slovakia Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Biofilms represent mixed communities present in a diverse range of environments; however, their utility as inoculants is less investigated. Our investigation was aimed towards in vitro development of biofilms using fungal mycelia ( Trichoderma viride ) as matrices and nitrogen-fixing and P-solubilizing bacteria as partners, as a prelude to their use as biofertilizers (biofilmed biofertilizers, BBs) and biocontrol agents for different crops. The most suitable media in terms of population counts, fresh mass and dry biomass for Trichoderma and Bacillus subtilis / Pseudomonas fluorescens was found to be Pikovskaya broth ± 1 % CaCO 3, while for Trichoderma and Azotobacter chroococcum , Jensen’s medium was most optimal . The respective media were then used for optimization of the inoculation rate of the partners in terms of sequence of addition of partners, fresh/dry mass of biofilms and population counts of partners for efficient film formation. Microscopic observations revealed significant differences in the progress of growth of biofilms and dual cultures. In the biofilms, the bacteria were observed growing intermingled within the fungal mycelia mat. Further, biofilm formation was compared under static and shaking conditions and the fresh mass of biofilms was higher in the former. Such biofilms are being further characterized under in vitro conditions, before using them as inoculants with crops. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0154-1 Authors Sodimalla Triveni, Division of Microbiology, Indian Agricultural Research Institute (IARI), New Delhi, 110012 India Radha Prasanna, Division of Microbiology, Indian Agricultural Research Institute (IARI), New Delhi, 110012 India Anil Kumar Saxena, Division of Microbiology, Indian Agricultural Research Institute (IARI), New Delhi, 110012 India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Antibodies have different avidities that can be evaluated using modified enzyme-linked immunosorbent assay (ELISA) techniques. We determined levels and avidities of antibodies to light (NFL) and medium (NFM) subunits of neurofilaments and tau protein in serum and cerebrospinal fluid (CSF) from 26 patients and anti-tau antibody levels and their avidities in 20 multiple sclerosis (MS) patients and 20 age- and sex-matched controls. Each sample was analyzed using both standard ELISA and also using a similar ELISA protocol with the addition of urea. The avidities of anti-neurocytoskeletal antibodies were higher in the CSF than those in serum (anti-NFL, p  〈 0.0001; anti-tau, p  〈 0.01; anti-NFM, n.s.). There was no relationship between avidities in serum and CSF for individual anti-neurocytoskeletal antibodies. We did not observe the relationship among the avidities of various anti-neurocytoskeletal antibodies. The avidities of anti-tau antibodies in the CSF were significantly higher in the MS patients than those in the controls ( p  〈 0.0001). The study demonstrates the differences in avidities of CSF or serum neurocytoskeletal antibodies measured as the urea resistance by ELISA method. Avidity determination of anti-neurocytoskeletal antibodies could contribute to the evaluation of the immunological status of patients. Content Type Journal Article Pages 1-5 DOI 10.1007/s12223-012-0105-x Authors L. Fialová, First Faculty of Medicine and General University Hospital, Institute of Medical Biochemistry and Laboratory Diagnostics, Charles University in Prague, Kateřinská 32, 121 08 Prague 2, Czech Republic J. Švarcová, First Faculty of Medicine and General University Hospital, Institute of Medical Biochemistry and Laboratory Diagnostics, Charles University in Prague, Kateřinská 32, 121 08 Prague 2, Czech Republic A. Bartos, Prague Psychiatric Center, Ústavní 91, 181 03, Bohnice, Prague 8, Czech Republic I. Malbohan, First Faculty of Medicine and General University Hospital, Institute of Medical Biochemistry and Laboratory Diagnostics, Charles University in Prague, Kateřinská 32, 121 08 Prague 2, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    A prerequisite for successful identification of anaerobic pathogenic bacteria from samples of clinical material is the method of cultivation. Currently, several methods of cultivation in anaerobic environment are used: cultivation in anaerobic box, anaerobic jar, and in nonrecurring cultivation system. Here, we determined the suitability of the above methods of cultivation using the estimation of the growth (diameters of colony size) of commonly isolated anaerobic pathogens ( Bacteroides fragilis , Clostridium difficile , and Clostridium perfringens ). The tested bacterial strains were exposed to atmospheric oxygen for various time periods and then they were cultivated using different anaerobic cultivation systems. Maximum growth differed, depending on the type of cultivation and the strain used. Thus, largest zone diameters, in the majority of measurements, were achieved in the anaerobic box. However, nonrecurring cultivation system seemed better in several cases; this applied to the cultivation of C . perfringens after 15, 30, and 60 min exposure to atmospheric oxygen as well as the cultivation of B . fragilis after 30 and 60 min of oxygen exposure. The cultivation in anaerobic box was the most convenient method for growth of C . difficile . In almost all cases, higher growth was observed in nonrecurring cultivation system than in the system of anaerobic jar. On the other hand, no significant differences were observed among these anaerobic cultivation systems which confirmed their applicability (taking into account some individual features concerning the optimization of cultivations) for identification of pathogenic anaerobes. Content Type Journal Article Pages 1-4 DOI 10.1007/s12223-012-0149-y Authors Ondřej Holý, Department of Preventive Medicine, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hnětovínská 3, 775 15 Olomouc, Czech Republic Dittmar Chmelař, Czech Anaerobic Bacteria Reference Laboratory, Faculty of Medicine, University of Ostrava, Syllabova 19, 703 00 Ostrava, Czech Republic Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    A halophilic isolate Thalassobacillus sp. LY18 producing extracellular amylase was isolated from the saline soil of Yuncheng Salt Lake, China. Production of the enzyme was synchronized with bacterial growth and reached a maximum level during the early stationary phase. The amylase was purified to homogeneity with a molecular mass of 31 kDa. Major products of soluble starch hydrolysis were maltose and maltotriose, indicating an α-amylase activity. Optimal enzyme activity was found to be at 70°C, pH 9.0, and 10 % NaCl. The α-amylase was highly stable over broad temperature (30–90°C), pH (6.0–12.0), and NaCl concentration (0–20 %) ranges, showing excellent thermostable, alkalistable, and halotolerant nature. The enzyme was stimulated by Ca 2+ , but greatly inhibited by EDTA, indicating it was a metalloenzyme. Complete inhibition by diethyl pyrocarbonate and β-mercaptoethanol revealed that histidine residue and disulfide bond were essential for enzyme catalysis. The surfactants tested had no significant effects on the amylase activity. Furthermore, it showed high activity and stability in the presence of water-insoluble organic solvents with log P ow  ≥ 2.13. Content Type Journal Article Pages 1-7 DOI 10.1007/s12223-012-0160-3 Authors Xin Li, Life Science College, Yuncheng University, Yuncheng, 044000 China Hui-Ying Yu, Life Science College, Yuncheng University, Yuncheng, 044000 China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    The pyrazosulfuron-ethyl-degrading bacterium, designated as CW17, was isolated from contaminated soil near the warehouse of the factory producing pyrazosulfuron-ethyl in Changsha city, China. The strain CW17 was identified as Acinetobacter sp. based on analyses of 94 carbon source utilization or chemical sensitivity in Biolog microplates, conventional phenotypic characteristics, and 16S rRNA gene sequencing. When pyrazosulfuron-ethyl was provided as the sole carbon source, the effects of pyrazosulfuron-ethyl concentration, pH, and temperature on biodegradation were examined. The degradation rates of pyrazosulfuron-ethyl at initial concentrations of 5.0, 20.0, and 50.0 mg/L were 48.0%, 77.0%, and 32.6%, respectively, after inoculation for 7 days. The growth of the strain was inhibited at low pH buffers. The chemical degradation occurs much faster at low pH than at neutral and basic pH conditions. The degradation rate of pyrazosulfuron-ethyl at 30°C was faster than those at 20 and 37°C by CW17 strains. Two metabolites of degradation were analyzed by liquid chromatography–mass spectroscopy (LC/MS). Based on the identified products, strain CW17 seemed to be able to degrade pyrazosulfuron-ethyl by cleavage of the sulfonylurea bridge. Content Type Journal Article Pages 1-9 DOI 10.1007/s12223-012-0107-8 Authors Yanhui Wang, Department of Pesticide Science, Hunan Agricultural University, 410128 Changsha, People’s Republic of China Liangwei Du, College of Chemistry and Chemical Engineering, Guangxi University, 530004 Nanning, Guangxi, People’s Republic of China Yingxi Chen, Department of Pesticide Science, Hunan Agricultural University, 410128 Changsha, People’s Republic of China Xiaoliang Liu, Institute of Pesticide and Environmental Toxicology, Guangxi University, 530004 Nanning, Guangxi, People’s Republic of China Xiaomao Zhou, Department of Pesticide Science, Hunan Agricultural University, 410128 Changsha, People’s Republic of China Huihua Tan, Institute of Pesticide and Environmental Toxicology, Guangxi University, 530004 Nanning, Guangxi, People’s Republic of China Lianyang Bai, Department of Pesticide Science, Hunan Agricultural University, 410128 Changsha, People’s Republic of China Dongqiang Zeng, Institute of Pesticide and Environmental Toxicology, Guangxi University, 530004 Nanning, Guangxi, People’s Republic of China Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632

Description:    Fifty fluorescent pseudomonads were isolated from rhizospheric soil of green gram from nearby area of Kaziranga, Assam, India and assayed for their extracellular proteinase production. Out of these isolates, 20 were found to be prominent in proteinase production. Genetic diversity of the 20 isolates were analyzed through BOX-PCR fingerprinting and 16S rDNA-RFLP along with three reference strains, viz., Pseudomonas fluorescens (NCIM2099 T ), Pseudomonas aureofaciens (NCIM2026 T ), and Pseudomonas aeruginosa (MTCC2582 T ). BOX-PCR produced two distinct clusters at 56% similarity coefficient and seven distinct BOX profiles. 16S rDNA-RFLP with three tetra-cutters restriction enzymes ( Hae III, Alu I, and Msp I) revealed two major clusters A and B; cluster A contained only single isolate FPS9 while the rest of 22 isolates belonged to the cluster B. Based on phenotypic characters and 16S rDNA sequence similarity, all the eight highly proteinase-producing strains were affiliated with P. aeruginosa . The proteinase was extracted from two most prominent strains (KFP1 and KFP2), purified by a three-step process involving (NH 4 ) 2 SO 4 precipitation, gel filtration, and ion exchange chromatography. The enzyme had an optimal pH of 8.0 and exhibit highest activity at 60°C and 37°C by KFP1 and KFP2 respectively. The specific activities were recorded as 75,050 (for KFP1) and 81,320 U/mg (for KFP2). The purified enzyme was migrated as a single band on native and SDS-PAGE with a molecular mass of 32 kDa. Zn 2+ , Cu 2+ , and Ni 2+ ion inhibited the enzyme activity. Enzyme activity was also inhibited by EDTA established as their metallo-proteinase nature. Content Type Journal Article Pages 1-9 DOI 10.1007/s12223-012-0097-6 Authors Rupak K. Sarma, Biotechnology Division, CSIR-North East Institute of Science and Technology, Jorhat, 785006 Assam, India Rajal Debnath, Biotechnology Division, CSIR-North East Institute of Science and Technology, Jorhat, 785006 Assam, India Ratul Saikia, Biotechnology Division, CSIR-North East Institute of Science and Technology, Jorhat, 785006 Assam, India Pratap J. Handique, Department of Biotechnology, Gauhati University, Guwahati, 781014 Assam, India Tarun C. Bora, Biotechnology Division, CSIR-North East Institute of Science and Technology, Jorhat, 785006 Assam, India Journal Folia Microbiologica Online ISSN 1874-9356 Print ISSN 0015-5632