SciCombinator

Discover the most talked about and latest scientific content & concepts.

Journal: World journal of microbiology & biotechnology

167

Disease-causing bacteria of the genus Aeromonas are able to adhere to pipe materials, colonizing the surfaces and forming biofilms in water distribution systems. The aim of our research was to study how the modification of materials used commonly in the water industry can reduce bacterial cell attachment. Polyvinyl chloride and silicone elastomer surfaces were activated and modified with reactive organo-silanes by coupling or co-crosslinking silanes with the native material. Both the native and modified surfaces were tested using the bacterial strain Aeromonas hydrophila, which was isolated from the Polish water distribution system. The surface tension of both the native and modified surfaces was measured. To determine cell viability and bacterial adhesion two methods were used, namely plate count and luminometry. Results were expressed in colony-forming units (c.f.u.) and in relative light units (RLU) per cm(2). Almost all the chemically modified surfaces exhibited higher anti-adhesive and anti-microbial properties in comparison to the native surfaces. Among the modifying agents examined, poly[dimethylsiloxane-co-(N,N-dimethyl-N-n-octylammoniopropyl chloride) methylsiloxane)] terminated with hydroxydimethylsilyl groups (20 %) in silicone elastomer gave the most desirable results. The surface tension of this modifier, was comparable to the non-polar native surface. However, almost half of this value was due to the result of polar forces. In this case, in an adhesion analysis, only 1 RLU cm(-2) and less than 1 c.f.u. cm(-2) were noted. For the native gumosil, the results were 9,375 RLU cm(-2) and 2.5 × 10(8) c.f.u. cm(-2), respectively. The antibacterial activity of active organo-silanes was associated only with the carrier surface because no antibacterial compounds were detected in liquid culture media, in concentrations that were able to inhibit cell growth.

Concepts: Archaea, Bacteria, Microbiology, Materials science, Bacterial cell structure, Microbiology terms, Silicone rubber, Aeromonas

28

Lindane is a notorious organochlorine pesticide due to its high toxicity, persistence in the environment and its tendency to bioaccumulate. A yeast strain isolated from sorghum cultivation field was able to use lindane as carbon and energy source under aerobic conditions. With molecular techniques, it was identified and named as Rhodotorula strain VITJzN03. The effects of nutritional and environmental factors on yeast growth and the biodegradation of lindane was investigated. The maximum production of yeast biomass along with 100 % lindane mineralization was noted at an initial lindane concentration of 600 mg l(-1) within a period of 10 days. Lindane concentration above 600 mg l(-1) inhibited the growth of yeast in liquid medium. A positive relationship was noted between the release of chloride ions and the increase of yeast biomass as well as degradation of lindane. The calculated degradation rate and half life of lindane were found to be 0.416 day(-1) and 1.66 days, respectively. The analysis of the metabolites using GC-MS identified the formation of seven intermediates including γ-pentachlorocyclohexane(γ-PCCH), 1,3,4,6-tetrachloro-1,4-cyclohexadiene(1,4-TCCHdiene), 1,2,4-trichlorobenzene (1,2,4 TCB), 1,4-dichlorobenzene (1,4 DCB), chloro-cis-1,2-dihydroxycyclohexadiene (CDCHdiene), 3-chlorocatechol (3-CC) and maleylacetate (MA) derivatives indicating that lindane degradation follows successive dechlorination and oxido-reduction. Based on the results of the present study, the possible pathway for lindane degradation by Rhodotorula sp. VITJzN03 has been proposed. To the best of our knowledge, this is the first report on lindane degradation by yeast which can serve as a potential agent for in situ bioremediation of medium to high level lindane-contaminated sites.

Concepts: Metabolism, Environment, Yeast, Ion, In situ, Chloride, Bioremediation, Rhodotorula

28

The current study aimed to test the hypothesis that both land-use change and soil type are responsible for the major changes in the fungal and archaeal community structure and functioning of the soil microbial community in Brazilian Pampa biome. Soil samples were collected at sites with different land-uses (native grassland, native forest, Eucalyptus and Acacia plantation, soybean and watermelon field) and in a typical toposequence in Pampa biome formed by Paleudult, Albaqualf and alluvial soils. The structure of soil microbial community (archaeal and fungal) was evaluated by ribosomal intergenic spacer analysis and soil functional capabilities were measured by microbial biomass carbon and metabolic quotient. We detected different patterns in microbial community driven by land-use change and soil type, showing that both factors are significant drivers of fungal and archaeal community structure and biomass and microbial activity. Fungal community structure was more affected by land-use and archaeal community was more affected by soil type. Irrespective of the land-use or soil type, a large percentage of operational taxonomic unit were shared among the soils. We accepted the hypothesis that both land-use change and soil type are drivers of archaeal and fungal community structure and soil functional capabilities. Moreover, we also suggest the existence of a soil microbial core.

Concepts: Archaea, Bacteria, Eukaryote, Ribosomal RNA, Soil, Steppe, Prairie, Alluvium

28

Amoebic liver abscess (ALA) is the most common extra intestinal manifestation of invasive amoebiasis caused by Entamoeba histolytica. The lack of early and accurate diagnostic test to differentiate various causes of liver abscess necessitates more reliable laboratory diagnostic test. The present study was conducted to assess the applicability of Loop-Mediated Isothermal Amplification (LAMP) assay for detection of E. histolytica in ALA cases. Fifty patients (n = 50) with clinical suspicion of ALA were enrolled in the study. All the clinical samples were subjected to conventional PCR assay. LAMP assay was standardized and the results were compared with that of PCR assay. Out of fifty pus samples thirty-six (72 %, 36/50) were positive for E. histolytica with conventional PCR assay and forty-one (82 %, 41/50) were positive by LAMP assay. Thus, five additional positive cases, missed by conventional PCR were positive with LAMP assay. Apart from rapidity, operational simplicity of LAMP assay high specificity and sensitivity, one-step amplification, higher yield and immediate visual detection may serve as a better diagnostic tool for diagnosis of ALA.

Concepts: Diagnosis, Type I and type II errors, Sensitivity and specificity, Entamoeba histolytica, Amoebic dysentery, Amoebozoa, Entamoeba, Liver abscess

28

Low ethanol yields and poor yeast viability were investigated at a continuous ethanol production corn wet milling facility. Using starch slurries and recycle streams from a commercial ethanol facility, laboratory hydrolysates were prepared by reproducing starch liquefaction and saccharification steps in the laboratory. Fermentations with hydrolysates prepared in the laboratory were compared with plant hydrolysates for final ethanol concentrations and total yeast counts. Fermentation controls were prepared using hydrolysates (plant and laboratory) that were not inoculated with yeast. Hydrolysates prepared in the laboratory resulted in higher final ethanol concentrations (15.8 % v/v) than plant hydrolysate (13.4 % v/v). Uninoculated controls resulted in ethanol production from both laboratory (12.2 % v/v) and plant hydrolysates (13.7 % v/v), indicating the presence of a contaminating microorganism. Yeast colony counts on cycloheximide and virginiamycin plates confirmed the presence of a contaminant. DNA sequencing and fingerprinting studies also indicated a number of dissimilar communities in samples obtained from fermentors, coolers, saccharification tanks, and thin stillage.

Concepts: Organism, Water, Ethanol, Yeast, Starch, Brewing, Fermentation, Ethanol fuel

28

Wine colour, phenolics and volatile fermentation-derived composition are the quintessential elements of a red wine. Many viticultural and winemaking factors contribute to wine aroma and colour with choice of yeast strain being a crucial factor. Besides the traditional Saccharomyces species S. cerevisiae, S. bayanus and several Saccharomyces interspecific hybrids are able to ferment grape juice to completion. This study examined the diversity in chemical composition, including phenolics and fermentation-derived volatile compounds, of an Australian Cabernet Sauvignon due to the use of different Saccharomyces strains. Eleven commercially available Saccharomyces strains were used in this study; S. cerevisiae (7), S. bayanus (2) and interspecific Saccharomyces hybrids (2). The eleven Cabernet Sauvignon wines varied greatly in their chemical composition. Nine yeast strains completed alcoholic fermentation in 19 days; S. bayanus AWRI 1375 in 26 days, and S. cerevisiae AWRI 1554 required 32 days. Ethanol concentrations varied in the final wines (12.7-14.2 %). The two S. bayanus strains produced the most distinct wines, with the ability to metabolise malic acid, generate high glycerol concentrations and distinctive phenolic composition. Saccharomyces hybrid AWRI 1501 and S. cerevisiae AWRI 1554 and AWRI 1493 also generated distinctive wines. This work demonstrates that the style of a Cabernet Sauvignon can be clearly modulated by choice of commercially available wine yeast.

Concepts: Ethanol, Yeast, Saccharomyces cerevisiae, Wine, Oenology, Fermentation, Cabernet Sauvignon, Winemaking

28

One of the most severe viral diseases of hill banana is caused by banana bunchy top virus (BBTV), a nanovirus transmitted by the aphid Pentalonia nigronervosa. In this study, we reported the Agrobacterium-mediated transformation on a highly valued hill banana cultivar Virupakshi (AAB) for resistance to BBTV disease. The target of the RNA interference (RNAi) is the rep gene, encoded by the BBTV-DNA1. In order to develop RNAi construct targeting the BBTV rep gene, the full-length rep gene of 870 bp was polymerase chain reaction amplified from BBTV infected hill banana sample DNA, cloned and confirmed by DNA sequencing. The partial rep gene fragment was cloned in sense and anti sense orientation in the RNAi intermediate vector, pSTARLING-A. After cloning in pSTARLING-A, the cloned RNAi gene cassette was released by NotI enzyme digestion and cloned into the NotI site of binary vector, pART27. Two different explants, embryogenic cells and embryogenic cell suspension derived microcalli were used for co-cultivation. Selection was done in presence of 100 mg/L kanamycin. In total, 143 putative transgenic hill banana lines were generated and established in green house condition. The presence of the transgenes was confirmed in the selected putative transgenic hill banana lines by PCR and reverse transcription PCR analyses. Transgenic hill banana plants expressing RNAi-BBTV rep were obtained and shown to resist infection by BBTV. The transformed plants are symptomless, and the replication of challenge BBTV almost completely suppressed. Hence, the RNAi mediating resistances were shown to be effective management of BBTV in hill banana.

Concepts: DNA, Gene expression, Polymerase chain reaction, Molecular biology, Enzyme, Virus, RNA, DNA replication

28

Loop-mediated isothermal amplification (LAMP), a novel method of gene amplification, was employed in this study for detecting Mycoplasma hyopneumoniae in the respiratory tract or lungs of swine. The pathogen can be detected in LAMP reactions containing as few as 10 fg purified target DNA (10 copies of M. hyopneumoniae genome) within 30 min, which was comparable to real-time PCR. After 30-min reaction at 63 °C, the addition of a certain amount of dye (SYBR Green I and hydroxyl naphthol blue at a proper ratio) into the LAMP reaction system makes the results easily determined as positive or negative by visual inspection. In addition, the LAMP was able to distinguish between M. hyopneumoniae and other closely-related mycoplasma strains, indicating a high degree of specificity. The LAMP assay was more simple and cheap, since the reaction could be completed under isothermal conditions and less laboratorial infrastructure are required. And, it was proven reliable for M. hyopneumoniae diagnosis of nasal swab and lung samples from the field.

Concepts: DNA, Bacteria, Polymerase chain reaction, Molecular biology, Respiratory system, Mycoplasma, Mollicutes, Mycoplasma hyopneumoniae

28

In order to better understand the differences in xylose metabolism between natural xylose-utilizing Pichia stipitis and metabolically engineered Saccharomyces cerevisiae, we constructed a series of recombinant S. cerevisiae strains with different xylose reductase/xylitol dehydrogenase/xylulokinase activity ratios by integrating xylitol dehydrogenase gene (XYL2) into the chromosome with variable copies and heterogeneously expressing xylose reductase gene (XYL1) and endogenous xylulokinase gene (XKS1). The strain with the highest specific xylose uptake rate and ethanol productivity on pure xylose fermentation was selected to compare to P. stipitis under oxygen-limited condition. Physiological and enzymatic comparison showed that they have different patterns of xylose metabolism and NADPH generation.

Concepts: Fungus, Yeast, Saccharomyces cerevisiae, Brewing, Beer, Yeasts, Saccharomycetes, Saccharomycetaceae

28

L-asparaginase from Cladosporium sp. grown on wheat bran by SSF was purified. Enzyme appeared to be a trimer with homodimer of 37 kDa and another 47 kDa amounting to total mass of 121 kDa as estimated by SDS-PAGE and 120 kDa on gel filtration column. The optimum temperature and pH of the enzyme were 30 °C and 6.3, respectively with Vmax of 4.44 μmol/mL/min and Km of 0.1 M. Substrate specificity studies indicated that, L-asparaginase has greater affinity towards L-asparagine with substrate hydrolysis efficiency (Vmax/Km ratio) eightfold higher than that of L-glutamine. L-asparaginase activity in presence of thiols studied showed decrease in Vmax and increase in Km, indicating nonessential mode of inactivation. Among the thiols tested, β-mercaptomethanol, exerted inhibitory effect, suggesting a critical role of disulphide linkages in maintaining a suitable conformation of the enzyme. Metal ions such as Ca(2+), Co(2+), Cu(2+), Mg(2+), Na(+), K(+) and Zn(2+) significantly affected enzyme activity whereas presence of Fe(3+), Pb(2+) and KI stimulated the activity. Detergents studied also enhanced L-asparaginase activity. In-vitro half-life of purified L-asparaginase in mammalian blood serum was 93.69 h. The enzyme inhibited acrylamide formation in potato chips by 96 % making it a potential candidate for food industry to reduce acrylamide content in starchy fried food commodities.

Concepts: Molecular biology, Enzyme, Enzyme substrate, Wheat, Starch, PH, Potato, Acrylamide