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Vol 85, No 1 (2016)

Reviews

Role of neuromediators in the functioning of the human microbiota: “Business talks” among microorganisms and the microbiota-host dialogue

Oleskin A.V., El’-Registan G.I., Shenderov B.A.

Abstract

Current concepts concerning the social behavior of microorganisms inhabiting the human gastrointestinal (GI) tract and their role in the formation of integrated supracellular structures and in intercellular communication in the host–microbiota system are reviewed. The analysis of the literature data and of the results obtained by the authors indicates an important role of neuromediators (biogenic amines, amino acids, peptides, and nitric oxide) in intra- and interspecies microbial communication, as well as in the microbiota–host dialogue. The role of this dialogue for human health, its effect on the human psyche and social behavior, and the possibility of construction of probiotic preparations with a target-oriented neurochemical effect are discussed.

Microbiology. 2016;85(1):1-22
pages 1-22 views

Experimental Articles

Role of thiol redox systems in Escherichia coli response to thermal and antibiotic stresses

Smirnova G.V., Lepekhina E.V., Muzyka N.G., Oktyabrsky O.N.

Abstract

Isogenic knockout mutants of Escherichia coli deficient in components of the glutathione and thioredoxin redox systems and growing at various temperatures (20–46°C) exhibited considerable differences in growth rate and survival, as well as in expression of the antioxidant genes. In the parental strain E. coli BW25113 (wt) treated with ciprofloxacin, ampicillin, or streptomycin, dependence of survival on growth temperature was a V-shaped curve with the maximum sensitivity within the range corresponding to high growth rates (40–44°C). Significant inverse correlation was observed between log CFU at different temperatures and specific growth rate prior to antibiotic addition. This applied to most of the mutants, which exhibited higher resistance to the three antibiotics tested at nonoptimal temperatures (20 and 46°C) than at 37 and 40°C. No correlation was found between resistance to stress and antibiotics and expression of the antioxidant genes. The role of global regulators ppGpp and σs in E. coli resistance to antibiotics and nonoptimal temperatures was shown.

Microbiology. 2016;85(1):23-32
pages 23-32 views

Survey on proteolytic activity and diversity of proteinase genes in mesophilic lactobacilli

Vukotić G., Strahinić I., Begović J., Lukić J., Kojić M., Fira D.

Abstract

Lactocepins or CEPs are large cell wall bound extracellular proteinases of lactic acid bacteria, involved in protein breakdown and utilization. They are responsible for many health-promoting traits of food products fermented with these organisms, but also essential for probiotic effects of certain strains. Different mesophilic strains selected within the species Lactobacillus zeae, Lb. casei, Lb. rhamnosus, and Lb. plantarum were analyzed for their proteolytic activity towards main fractions of milk proteins—caseins and whey proteins. The strains showing excellent proteolytic features were further examined for presence of corresponding proteinase gene(s). It was found that Lb. zeae LMG17315 possessed catalytic domains of three distinct proteinase genes, unique feature in Lb. casei group, which are similar but not identical to previously characterized prtP and prtR genes. Lb. casei neotype strain ATCC393 was also analysed and based on obtained results its reclassification in taxon Lb. zeae is supported. In addition, we report catalytic domain of prtR-type gene in Lb. plantarum LMG9208, which is first such report in this species, and it is first time that this gene is reported outside Lb. casei group.

Microbiology. 2016;85(1):33-41
pages 33-41 views

Effect of mutations in extracellular nuclease on the characteristics of the pigmented and nonpigmented Serratia marcescens strains

Nizamutdinova E.K., Shirshikova T.V., Mardanova A.M., Sharipova M.R., Bogomol’naya L.M.

Abstract

Comparative characterization of the pigmented and nonpigmented Serratia marcescens strains and their extracellular nuclease mutants was carried out. Biomass accumulation by the mutant strains decreased on average by 20%, while proteolytic activity of the culture liquid was 4–5 times lower than in the case of the wild type strains. The mutants with impaired extracellular nuclease genes exhibited higher sensitivity to reactive oxygen species. Comparative analysis of motility of the strains revealed the highest flagellar activity in the wild type nonpigmented strain, while the cells of its mutant completely lost this feature.

Microbiology. 2016;85(1):42-46
pages 42-46 views

Determination of endoglucanase activity of paper decaying fungi from an old library at the ancient Medina of Fez

El Bergadi F., Laachari F., Sadiki M., Elabed S., Iraqui M.H., Ibnsouda S.K.

Abstract

Paper from an ancient library of the cultural city of Fez (Morocco) is exposed to rapid deterioration by variety of microorganisms, especially cellulolytic fungi. For this, ten isolates fungi previously isolated from historical biodeteriorated paper were screened for their ability to produce endoglucanase (CMCase), amylase, polygalacturonase and ligninase enzymes. The CMCase activity of cellulolytic strains was essayed in liquid media at 25°C for 10 days. Influence of temperature and pH were assessed for the production of CMCase by all the fungus isolated from decaying paper. The research findings from the present study demonstrate that all the tested isolates had cellulase, amylase, pectinase and ligninase activities. It was found that Mucor racemosus PF15, Aspergillus niger, and Aspergillus oryzae exhibited the maximum endoglucanase activity in liquid medium (0.256, 0.236, and 0.216 UI/mL in descending order) for six days. Temperature profiling revealed optimum endoglucanase activity at 25 and 30°C. Maximum activity was observed at pH 5 and pH 6.

Microbiology. 2016;85(1):47-55
pages 47-55 views

Isolation and characterization of biosurfactant producing Bacillus sp. from diesel fuel-contaminated site

Yadav A.K., Manna S., Pandiyan K., Singh A., Kumar M., Chakdar H., Kashyap P.L., Srivastava A.K.

Abstract

Among hydrocarbon pollutants, diesel oil is a complex mixture of alkanes and aromatic compounds which are often encountered as soil contaminants leaking from storage tanks and pipelines or as result of accidental spillage. One of the best ecofriendly approaches is to restore contaminated soil by using microorganisms able to degrade those toxic compounds in a bioremediation process. In the present study, nineteen bacteria were isolated by enrichment culture technique from diesel spilled soil collected from electric generator shed of NBAIM, Mau. All the isolates were subjected to screening for lipase production and twelve isolates were found to be positive for lipase. When the isolates were screened for biosurfactant production using CTAB-methylene blue agar plates, only one isolate viz. 2NBDSH3 was found positive which was found to be phylogenetically closely related with Bacillus flexus. Despite having low emulsification index, the bacterium could degrade 88.6% of diesel oil in soil. Biosurfactant from the isolate was extracted and characterized through infra-red spectroscopy which indicated its possible lipopeptide nature which was further supported by strong absorption in UV range in the UV-Vis spectrum. The results of the present study indicated that the isolate either does not produce any bioemulsifier or produces very low amount of emulsifier rather it produces a lipopeptide biosurfactant which helps in degradation of diesel oil by lowering the surface tension. The bacterium thus isolated and characterized can serve as a promising solution for ecofriendly remediation of bacterium diesel contaminated soils.

Microbiology. 2016;85(1):56-62
pages 56-62 views

Production of gold nanoparticles by biogenesis using bacteria

Radtsig M.A., Koksharova O.A., Nadtochenko V.A., Khmel’ I.A.

Abstract

Diazotrophic cyanobacteria Anabaena sp. PCC 7120, four Nostoc strains, and two Azotobacter species (A. vinelandii and A. chroococcum) were found to produce gold nanoparticles (GNP) under nitrogen fixation conditions. GNP biogenesis occurred at AuHCl4 concentrations from 0.1 to 1 mM. In the cultures of unicellular cyanobacteria Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803 incapable of nitrogen fixation, no GNP were formed at the same concentrations of gold salts. The plasmon resonance band peak was located at 552 nm. This position is characteristic of spherical GNP 10 to 30 nm in size. Small amounts of GNP were also formed in the culture liquid supernatants of the tested nitrogen-fixing bacteria at AuHCl4 concentrations from 0.25 to 0.5 mM.

Microbiology. 2016;85(1):63-70
pages 63-70 views

An efficient method for genetic certification of Bacillus subtilis strains, prospective producers of biopreparations

Terletsky V.P., Tyshchenko V.I., Novikova I.I., Boikova I.V., Tyulebaev S.D., Shakhtamirov I.Y.

Abstract

Genetic certification of commercial strains of bacteria antagonistic to phytopathogenic microorganisms guarantees their unequivocal identification and confirmation of safety. In Russia, unlike EU countries, genetic certification of Bacillus subtilis strains is not used. Based on the previously proposed double digestion selective label (DDSL) fingerprinting, a method for genetic identification and certification of B. subtilis strains was proposed. The method was tested on several strains differing in their physiological and biochemical properties and in the composition of secondary metabolites responsible for the spectrum of antibiotic activity. High resolving power of this approach was shown. Optimal restriction endonucleases (SgsI and Eco32I) were determined and validated. A detailed protocol for genetic certification of this bacterial species was developed. DDSL is a universal method, which may be adapted for genetic identification and certification of other bacterial species. Keywords: genotyping, restriction

Microbiology. 2016;85(1):71-76
pages 71-76 views

New strains of an aerobic anoxygenic phototrophic bacterium Porphyrobacter donghaensis isolated from a Siberian thermal spring and a weakly mineralized lake

Nuyanzina-Boldareva E.N., Akimov V.N., Takaichi S., Gorlenko V.M.

Abstract

A strain of aerobic anoxygenic phototrophic bacteria (AAPB) isolated from the surface of a cyanobacterial mat of an Eastern Siberian thermal spring (40°C) and designated Se-4 was identified as Porphyrobacter donghaensis according to its 16S rRNA gene sequence. A DNA–DNA hybridization level of 95% was determined between strain Se-4 and the type strain of this species, SW-132T. The isolate was an obligate aerobe, forming orange round colonies on solid media, which turn red in the course of growth. The cells were motile rods capable of branching. The cells divided by uniform fission by constriction. Optimal growth was observed at pH 7.5 and NaCl concentrations from 0 to 1 g/L. The pigments present were carotenoids and bacteriochlorophyll a. Another Porphyrobacter donghaensis strain, Noj-1, isolated from a purple mat developing on the surface of a coastal set-up in a steppe weakly mineralized (1.5 g/L) soda lake Nozhii (Eastern Siberia) possessed similar characteristics. Thus, the AAPB species Porphyrobacter donghaensis was shown to occur, apart from marine environments (its known habitat), also in weakly mineralized soda lakes and freshwater thermal springs. Description of the species Porphyrobacter donghaensis was amended.

Microbiology. 2016;85(1):77-86
pages 77-86 views

Analysis of proliferation and survival of agrobacteria after inoculation of maize pistil filaments

Volokhina I.V., Velikov V.A., Moiseeva E.M., Chumakov M.I.

Abstract

While the authors have previously developed a method of pistil filament treatment with Agrobacterium cells during blossoming for the transformation of maize generative cells, the mechanism for bacterial T-DNA penetration into the embryo sac remained unknown. This article analyzes the possibility of agrobacterial penetration into the maize embryo via pollen tubes. Microbiological, PCR, and GUS techniques were used to confirm that agrobacteria could spread for up to 20 cm from the site of inoculation and were detected in maize embryo tissues as early as 24 h after inoculation, while they were not revealed after 5–13 days.

Microbiology. 2016;85(1):87-92
pages 87-92 views

Dynamics of purple sulfur bacteria in a meromictic saline Lake Shunet (Khakassia, Siberia) in 2007–2013

Rogozin D.Y., Zykov V.V., Tarnovskii M.O.

Abstract

According to the results of seasonal monitoring, in 2007–2013 purple sulfur bacteria morphologically similar to Thiocapsa sp. Shira_1 (AJ633676 in EMBL/GenBank) predominated in the anoxygenic phototrophic community of the water column of the meromictic Lake Shira (Khakassia, Siberia). No pronounced seasonal periodicity in the total cell number in the water column was revealed during the period of observation. In some years cell number during the period when the lake was covered with ice was reliably higher than in summer. The absence of seasonal periodicity was probably due to the low amplitude of seasonal variations in temperature and illumination in the redox zone, resulting from its relatively deep location (12–16 m). The year-to-year dynamics was characterized by a reliable decrease of the total cell number in 2009–2010 and maxima in 2007 and 2011–2012. Canonical correlation analysis revealed that water temperature in the redox zone was the best predictor of the PSB abundance in Lake Shira. Water temperature, in turn, depended on the depth of mixing of the water column. Intense mixing in 2009–2011 was probably responsible for decreased PSB abundance in the lake. On the other hand, the absence of deep winter mixing, resulting in stable conditions in the chemocline, favored the preservation of relatively high PSB biomass. Prediction of circulation depth, which depends mainly on the weather conditions and dynamics of the water level, is required for prediction of PSB abundance in Lake Shira. These results may be useful for paleolimnological reconstructions of the history of the lake based on the remnants of purple sulfur bacteria in bottom sediments.

Microbiology. 2016;85(1):93-101
pages 93-101 views

Characterization of the structure of the prokaryotic complex of Antarctic permafrost by molecular genetic techniques

Manucharova N.A., Trosheva E.V., Kol’tsova E.M., Demkina E.V., Karaevskaya E.V., Rivkina E.M., Mardanov A.V., El’-Registan G.I.

Abstract

A prokaryotic mesophilic organotrophic community responsible for 10% of the total microbial number determined by epifluorescence microscopy was reactivated in the samples of Antarctic permafrost retrieved from the environment favoring long-term preservation of microbial communities (7500 years). No culturable forms were obtained without resuscitation procedures (CFU = 0). Proteobacteria, Actinobacteria, and Firmicutes were the dominant microbial groups in the complex. Initiation of the reactivated microbial complex by addition of chitin (0.1% wt/vol) resulted in an increased share of metabolically active biomass (up to 50%) due to the functional domination of chitinolytics caused by the target resource. Thus, sequential application of resuscitation procedures and initiation of a specific physiological group (in this case, chitinolytics) to a permafrost-preserved microbial community made it possible to reveal a prokaryotic complex capable of reversion of metabolic activity (FISH data), to determine its phylogenetic structure by metagenomic analysis, and to isolate a pure culture of the dominant microorganism with high chitinolytic activity.

Microbiology. 2016;85(1):102-108
pages 102-108 views

Gut bacterial diversity of farmed sea cucumbers Apostichopus japonicus with different growth rates

Sha Y., Liu M., Wang B., Jiang K., Sun G., Wang L.

Abstract

Farmed sea cucumbers (Apostichopus japonicus) of the same age displayed significantly different body sizes and weights under the same farming conditions. To examine the gut bacterial diversity of sea cucumbers and whether the growth differences between them were related to intestinal microbiota, the bacterial species from intestinal samples of 30 farmed A. japonicus from the same tank (15 large sea cucumbers vs. 15 small sea cucumbers) were characterized based on 16S rRNA gene analysis by means of high throughout sequencing. The results showed that bacterial phylotypes in both sizes of sea cucumbers were closely related to Proteobacteria, Verrucomicrobia, Bacteroidetes, Actinobacteria, Firmicutes, Cyanobacteria, Planctomycete, and Spirochaetes, of which Proteobacteria were predominant (>50%). There were no significant differences in the relative abundances of each bacterial phylotype between the two groups, except for Actinobacteria (P < 0.05). In addition, different species uniquely belonging to all three tested samples in the large group and the small group were found. It was interestingly that Vibrio were absent from both groups. It is likely that the differences in the abundances of Actinobacteria and different species in the two groups may be related to their remarkable disparities in body sizes.

Microbiology. 2016;85(1):109-115
pages 109-115 views

Characterization of the gut bacterial community of the Japanese sea cucumber Apostichopus japonicus

Bogatyrenko E.A., Buzoleva L.S.

Abstract

Comparative analysis of the composition and abundance of the gut bacterial community of the Japanese sea cucumber (trepang) Apostichopus japonicus and the sediment from its habitat was carried out. In spite of the presence of strictly aerobic bacteria in the sediments, gut microbiota of this holothurian was shown to be formed by facultative anaerobes from the environment. Irrespective of the geographical location of the habitat, the sea cucumber gut community had a constant composition, considered its normal microflora. Capacity of bacteria isolated from the Japanese sea cucumber for decomposition of starch, sodium alginate, chitin, chondroitin sulfate, Tweens, olive oil, casein, and gelatin was studied. Various degrees of enzymatic activity were shown for 33% of the studied bacterial strains, which indicates a considerable role of the trepang gut microflora in processing and assimilation of organic matter arriving with food.

Microbiology. 2016;85(1):116-123
pages 116-123 views

Yeast communities of Formica aquilonia colonies

Maksimova I.A., Glushakova A.M., Kachalkin A.V., Chernov I.Y., Panteleeva S.N., Reznikova Z.I.

Abstract

Yeast abundance and species diversity in the colonies of Formica aquilonia ants in birch–pine grass forest near Novosibirsk, Russia, were studied. The average yeast number in the anthill material was 103–104 CFU/g, reaching 105 CFU/g in the hatching chambers. Typical litter species (Trichosporon moniliiforme and Cystofilobasidium capitatum) were predominant in soil and litter around the anthills. Apart from these species, ascomycete species of the family Debaryomycetaceae, Debaryomyces hansenii, and Schwanniomyces vanrijiae were predominant in the anthill material. Yeast population of the ant’ bodies consisted exclusively of the members of the last two species. Thus, highly specific yeast communities formed in the colonies of Formica aquilonia ants differ from the communities of surrounding soil. These differences are caused by environment-forming activity of the ants.

Microbiology. 2016;85(1):124-129
pages 124-129 views

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