The characterization of oil-degrading microorganisms from lubricating oil contaminated (scale) soil

AIMS: To isolate and characterize oil-degrading microorganisms from contaminated (scale) soil. METHODS AND RESULTS: Oil-degrading microorganisms were isolated from enrichment cultures of scale soil. Each isolate was identified using 16S rDNA gene and oil degradability was determined on both unused and used lubricating oil. The weight of the extracted remaining oil revealed that most isolates degraded unused lubricating oil more than used lubricating oil. Chemical composition of oil analysed by TLC-FID and GC-MS demonstrated that Nocardia simplex W9 degraded used oil the best, and resulted in a decrease in saturates, aromatics and resins to 52.46, 38.13 and 18.81%, respectively. CONCLUSIONS:Nocardia simplex W9 is the best degrader, among all the isolates, on both used and unused lubricating oil. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of Nocardia simplex W9 in scale soil enables iron to be recycled by biodegradation.     

Selection and characterization of active psychrotrophic microbial oil-degrading microorganisms

The ability of 96 microbial strains degrading oil and 32 strains degrading polycyclic aromatic hydrocarbons (PAHs) to consume diesel fuel and oil at 4–6 and 24°C and at elevated NaCl concentrations was studied. The temperature range, salt tolerance, ability to produce biosurfactants, range of substrates, and antibiotic resistance were determined. The eleven most active oil-degrading and PAH-degrading strains were genotyped by a polymerase chain reaction with BoxA1R primers and a restriction analysis of ribosomal DNA amplicons. For six strains, the degree of oil degradation at 4–6°C was higher than at 24°C. For the most active strains, the degree of oil degradation in liquid mineral medium ranged from 15 to 26% at 24°C and from 28 to 47% at 4–6°C. An association of six of the strains degraded the oil by 46% at 24°C.     

Study of the stability association of oil-degrading microorganisms in an open system

Investigation of the stability of an association of active hydrocarbon-oxidizing microorganisms grown on diesel fuel showed that not all members of the association were competitive in an open flow system. The abundance of some strains considerably decreased during cultivation. One of the strain groups had consistently high cell titers, both in the medium and on a support. This prompts the authors to use these strains as degraders of diesel fuel.     

Presence of microorganisms in hemolymph of the horseshoe crab Limulus polyphemus.

Hemolymph samples obtained from Limulus polyphemus at the time of collection and after a 1-week holding period exhibited a significant increase in bacterial levels. No differences were observed in the ability of amoebocyte lysate, prepared from these same samples, to gel in the presence of lipopolysaccharide.     

Biochemistry of the initial steps of mycothiol biosynthesis

Mycothiol is the major thiol produced by mycobacteria and is required for growth of Mycobacterium tuberculosis. The final three steps in the biosynthesis of mycothiol have been fully elucidated but the initial steps have been unclear. A glycosyltransferase, MshA, is required for production of the mycothiol precursor, 1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol, but its substrates and immediate products were unknown. In this study, we show that the N-acetylglucosamine donor is UDP-N-acetylglucosamine and that the N-acetylglucosamine acceptor is 1L-myo-inositol 1-phosphate. The reaction generates UDP and 1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol 3-phosphate. Using cell-free extracts of M. smegmatis mc(2)155, little activity was obtained with myo-inositol, 1D-myo-inositol 1-phosphate, or myo-inositol 2-phosphate as the N-acetylglucosamine acceptor. A phosphatase, designated MshA2, is required to dephosphorylate 1-O-(2-acetamido-2-deoxy-alpha-glucopyranosyl)-D-myo-inositol 3-phosphate to produce 1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol. The latter is deacetylated, ligated with cysteine, and the cysteinyl amino group acetylated by acetyl-CoA to complete the mycothiol biosynthesis pathway. Uptake and concentration of myo-[14C]inositol is rapid in Mycobacterium smegmatis and leads to production of radiolabeled inositol 1-phosphate and mycothiol. This demonstrates the presence of a myo-inositol transporter and a kinase that generates 1L-myo-inositol 1-phosphate. The biochemical pathway of mycothiol biosynthesis is now fully elucidated.     

Cytoskeletons of ADP- and thrombin-stimulated blood platelets. Presence of a caldesmon-like protein, alpha-actinin and gelsolin at different steps of the stimulation

Comparative analyses of the cytoskeletons of resting and stimulated platelets point out the involvement of a 79 kDa polypeptide in the activation step and its increased incorporation during aggregation. It appears as a doublet and cross-reacts with an antibody to chicken gizzard caldesmon, whereas no 150 kDa immunoreactive form was detected. alpha-Actinin and gelsolin were detected only in the aggregation step.     

Structural insights into initial and intermediate steps of the ribosome-recycling process

The ribosome-recycling factor (RRF) and elongation factor-G (EF-G) disassemble the 70S post-termination complex (PoTC) into mRNA, tRNA, and two ribosomal subunits. We have determined cryo-electron microscopic structures of the PoTC·RRF complex, with and without EF-G. We find that domain II of RRF initially interacts with universally conserved residues of the 23S rRNA helices 43 and 95, and protein L11 within the 50S ribosomal subunit. Upon EF-G binding, both RRF and tRNA are driven towards the tRNA-exit (E) site, with a large rotational movement of domain II of RRF towards the 30S ribosomal subunit. During this intermediate step of the recycling process, domain II of RRF and domain IV of EF-G adopt hitherto unknown conformations. Furthermore, binding of EF-G to the PoTC·RRF complex reverts the ribosome from ratcheted to unratcheted state. These results suggest that (i) the ribosomal intersubunit reorganizations upon RRF binding and subsequent EF-G binding could be instrumental in destabilizing the PoTC and (ii) the modes of action of EF-G during tRNA translocation and ribosome-recycling steps are markedly different.     

The sensitivity to antibiotics and chloramine B of opportunistic microorganisms isolated from patients

Strains of staphylococci, klebsiella, proteins, blue pus bacillus and enterobacteria resistant not only to antibiotics but also to chloramine, a disinfectant, were isolated from the test material samples obtained from patients with different purulent-inflammatory processes. The following strains are revealed among the test microorganisms: sensitive simultaneously to antibiotics and chloramine; sensitive to antibiotics but resistant to chloramine; sensitive to chloramine but resistant to antibiotics; simultaneously resistant both to antibiotics and chloramine.     

Intrinsic signals regulate the initial steps of myogenesis in vertebrates

In vertebrates, despite the evidence that extrinsic factors induce myogenesis in naive mesoderm, other experiments argue that the initiation of the myogenic program may take place independent of these factors. To resolve this discrepancy, we have re-addressed this issue, using short-term in vivo microsurgery and culture experiments in chick. Our results show that the initial expression of the muscle-specific markers Myf5 and MyoD is regulated in a mesoderm-autonomous fashion. The reception of a Wnt signal is required for MyoD, but not Myf5 expression; however, we show that the source of the Wnt signal is intrinsic to the mesoderm. Gain- and loss-of-function experiments indicate that Wnt5b, which is expressed in the presomitic mesoderm, represents the MyoD-activating cue. Despite Wnt5b expression in the presomitic mesoderm, MyoD is not expressed in this tissue: our experiments demonstrate that this is due to a Bmp inhibitory signal that prevents the premature expression of MyoD before somites form. Our results indicate that myogenesis is a multistep process which is initiated prior to somite formation in a mesoderm-autonomous fashion; as somites form, influences from adjacent tissues are likely to be required for maintenance and patterning of early muscles.     

The specific activity of a multicomponent vaccine made from the antigens of opportunistic microorganisms

Multicomponent vaccine prepared from the antigens of 4 representatives of opportunistic microflora possesses high specific activity. The passive hemagglutination (PHA) test with the use of associated diagnosticum showed that antibody titers in the sera of immunized rabbits increased 10- to 10(4)-fold in comparison with the titers observed prior to immunization. The PHA test with the use of the antigens contained in the vaccine revealed the accumulation of antibodies to each of the 4 components of the preparation in the blood sera of immunized rabbits. When stored at 4 degrees C, the vaccine was shown to retain its specific activity for 5 years (the term of observation).     

Presence of microorganisms in hemolymph of the horseshoe crab Limulus polyphemus

Hemolymph samples obtained from Limulus polyphemus at the time of collection and after a 1-week holding period exhibited a significant increase in bacterial levels. No differences were observed in the ability of amoebocyte lysate, prepared from these same samples, to gel in the presence of lipopolysaccharide.     

Electrophoretic mobility and hydrophobicity as a measured to predict the initial steps of bacterial adhesion.

The relationship between physiochemical surface parameters and adhesion of bacterial cells to negatively charged polystyrene was studied. Cell surface hydrophobicity and electrokinetic potential were determined by contact angle measurement and electrophoresis, respectively. Both parameters influence cell adhesion. The effect of the electrokinetic potential increases with decreasing hydrophobicity. Cell surface characteristics determining adhesion are influenced by growth conditions. At high growth rates, bacterial cells tend to become more hydrophobic. This fact can be of ecological significance for controlling the spread of bacteria throughout the environment.     

Experimental and clinical studies of drug regulation of the anti-lysozyme activity of microorganisms causing opportunistic infections

Experimental materials on choosing antibiotics for etiotropic therapy of opportunistic infections with an account of the regulating effect of the drugs on the ++anti-lysozyme activity of pathogens (the factor of intracellular parasitism) are presented. The in vitro data were applied to the clinical trials in 30 patients with chronic and acute pyelonephritis of the Proteus etiology and to 25 patients with chronic inflammatory diseases of Staphylococcus etiology. It was shown that the use of the antibiotics which lowered the ++anti-lysozyme activity of microorganisms promoted a more rapid disappearance of the disease clinical signs, increased 2- to 3-fold the terms of the remission and resulted in an increase in the number of the persons with complete remission (54.5 to 63.6 per cent) as compared to the use of the drugs which stimulated the pathogen property or were indifferent to it.     

Isolation and identification of crude oil-degrading yeast strains from Khafji oil field,saud Arabia

Twenty five yeasts isolated were isolated from Khurais oil field in Saudi Arabia and assayed to evaluate their biodegradability. Only five isolates (namely, A1, A2, A3, A4 and A5) showed potential use of oil as sole carbon source. During incubation period, highest growth rate were recorded for A1, A2 and A3 isolates. Low growth distinguished A4 isolate; A5 isolate could not degrade oil.Spectrophotometrical analysis for four yeast isolates biodegradation activities indicated that, A1 isolate was superior for oil degradation (61%) comparing with A4 isolate which reflected lowest degradation % (33%). A2 and A3 isolates showed moderate biodegradation activity (56 and 51% respectively).D1/D2 domain of the 26S rRNA gene sequence was used as molecular marker to identify five yeast isolates. After comparing 26S rRNA gene sequences of five yeast isolates with highly similarity isolates, five yeast isolates (A1, A2, A3, A4 and A5)were submitted to database as Candida tropicalis (MW488263), Candida tropicalis (MW488264), Rhodotorula mucilaginosa (MW488265) and Rhodosporidium toruloides (MW488266) respectively. Using OXF1/ACR1 primer, specific lipase gene amplicon with 250 bp were detected with in all four yeast isolates.     

Control of the initial steps in heme biosynthesis in free-livingRhizobium sp. by culture conditions

Cell-free extracts obtained from free-livingRhizobium sp. in early stationary phase had three times as much 5-aminolevulinate synthase activity as did similar extracts from log phase cells. The level of 5-aminolevulinate dehydratase was also elevated at this point. The presence of 0.1 mM hemin in the culture medium prevented the transitory increase in enzyme activities during this early stationary phase. The effect of hemin was counteracted by 1 mg bovine serum albumin per milliliter medium. This control of the development of 5-aminolevulinate synthase and 5-aminolevulinate dehydratase activities by free hemin suggests a mechanism by which heme and globin formation might be coordinated for the synthesis of leghemoglobin in legume root nodules.     

Immunostimulant preparation made from the antigens of opportunistic microorganisms in the treatment of inflammatory lung diseases

The results of the trial of an immunostimulating preparation, consisting of Klebsiella pneumoniae, Proteus, Escherichia coli and Staphylococcus antigenic complexes, on 20 patients with acute pulmonary abscess and bronchiectasis are presented. The preparation was introduced subcutaneously in 5 injections. The preparation was found to have low reactogenicity, and in the course of immunotherapy the manifestations of systemic and local reactions became considerably less pronounced. Immunotherapy produced a good curative effect, objectively manifested by a decrease in coughing and in the amount of sputum gradually changing its character. After the fourth and fifth injections the patients no longer ejected purulent sputum. Fluoroscopic examination revealed a considerable decrease in the size of the pathological focus. The preparation stimulated immunological reactions, and immunization resulted in a considerable increase in the titer of antibodies to all components of the combined preparation, as well as in an increase in the number of functionally active T-lymphocytes, in the blood of the patients.