Comparative Bioremediation of Tetradecane, Cyclohexanone and Cyclohexane by Filamentous Fungi from Polluted Habitats in Kazakhstan.

Studying the fates of oil components and their interactions with ecological systems is essential for developing comprehensive management strategies and enhancing restoration following oil spill incidents. The potential expansion of Kazakhstan's role in the global oil market necessitates the existence of land-specific studies that contribute to the field of bioremediation. In this study, a set of experiments was designed to assess the growth and biodegradation capacities of eight fungal strains sourced from Kazakhstan soil when exposed to the hydrocarbon substrates from which they were initially isolated. The strains were identified as Aspergillus sp. SBUG-M1743, Penicillium javanicum SBUG-M1744, SBUG-M1770, Trichoderma harzianum SBUG-M1750 and Fusarium oxysporum SBUG-1746, SBUG-M1748, SBUG-M1768 and SBUG-M1769 using the internal transcribed spacer (ITS) region. Furthermore, microscopic and macroscopic evaluations agreed with the sequence-based identification. Aspergillus sp. SBUG-M1743 and P. javanicum SBUG-M1744 displayed remarkable biodegradation capabilities in the presence of tetradecane with up to a 9-fold biomass increase in the static cultures. T. harzianum SBUG-M1750 exhibited poor growth, which was a consequence of its low efficiency of tetradecane degradation. Monocarboxylic acids were the main degradation products by SBUG-M1743, SBUG-M1744, SBUG-M1750, and SBUG-M1770 indicating the monoterminal degradation pathway through ß-oxidation, while the additional detection of dicarboxylic acid in SBUG-M1768 and SBUG-M1769 cultures was indicative of the fungus' ability to undertake both monoterminal and diterminal degradation pathways. F. oxysporum SBUG-M1746 and SBUG-M1748 in the presence of cyclohexanone showed a doubling of the biomass with the ability to degrade the substrate almost completely in shake cultures. F. oxysporum SBUG-M1746 was also able to degrade cyclohexane completely and excreted all possible metabolites of the degradation pathway. Understanding the degradation potential of these fungal isolates to different hydrocarbon substrates will help in developing effective bioremediation strategies tailored to local conditions.

Bacterial crude oil and polyaromatic hydrocarbon degraders from Kazakh oil fields as barley growth support.

Bacterial strains of the genera Arthrobacter, Bacillus, Dietzia, Kocuria, and Micrococcus were isolated from oil-contaminated soils of the Balgimbaev, Dossor, and Zaburunye oil fields in Kazakhstan. They were selected from 1376 isolated strains based on their unique ability to use crude oil and polyaromatic hydrocarbons (PAHs) as sole source of carbon and energy in growth experiments. The isolated strains degraded a wide range of aliphatic and aromatic components from crude oil to generate a total of 170 acid metabolites. Eight metabolites were detected during the degradation of anthracene and of phenanthrene, two of which led to the description of a new degradation pathway. The selected bacterial strains Arthrobacter bussei/agilis SBUG 2290, Bacillus atrophaeus SBUG 2291, Bacillus subtilis SBUG 2285, Dietzia kunjamensis SBUG 2289, Kocuria rosea SBUG 2287, Kocuria polaris SBUG 2288, and Micrococcus luteus SBUG 2286 promoted the growth of barley shoots and roots in oil-contaminated soil, demonstrating the enormous potential of isolatable and cultivable soil bacteria in soil remediation. KEY POINTS: • Special powerful bacterial strains as potential crude oil and PAH degraders. • Growth on crude oil or PAHs as sole source of carbon and energy. • Bacterial support of barley growth as resource for soil remediation.

Exploring Potential of Gellan Gum for Enhanced Oil Recovery.

Extensive laboratory and field tests have shown that the gelation response of gellan gum to saline water makes it a promising candidate for enhanced oil recovery (EOR). The objective of this mini-review is to evaluate the applicability of gellan gum in EOR and compare its efficiency to other precursors, in particular, hydrolyzed polyacrylamide (HPAM). At first, the "sol-gel" phase transitions of gellan gum in aqueous-salt solutions containing mono- and divalent cations are considered. Then the rheological and mechanical properties of gellan in diluted aqueous solutions and gel state are outlined. The main attention is paid to laboratory core flooding and field pilot tests. The plugging behavior of gellan in laboratory conditions due to "sol-gel" phase transition is discussed in the context of conformance control and water shut-off. Due to its higher strength, gellan gum gel provided ~6 times greater resistance to the flow of brine in a 1 mm-width fracture compared to HPAM gel. The field trials carried out in the injection and production wells of the Kumkol oilfield, situated in Kazakhstan, demonstrated that over 6 and 11 months, there was an incremental oil recovery of 3790 and 5890 tons, respectively. To put it into perspective, using 1 kg of dry gellan resulted in the incremental production of 3.52 m3 (or 22 bbls) of oil. The treatment of the production well with 1 wt.% gellan solution resulted in a considerable decrease in the water cut up to 10-20% without affecting the oil flow rate. The advantages and disadvantages of gellan compared to HPAM are analyzed together with the economic feasibility of gellan over HPAM. The potential for establishing gellan production in Kazakhstan is emphasized. It is anticipated that gellan gum, manufactured through fermentation using glucose-fructose syrup from Zharkent and Burunday corn starch plants, could be expanded in the future for applications in both the food industry and oil recovery.

Characterization of the Mycoremediation of n-Alkanes and Branched-Chain Alkanes by Filamentous Fungi from Oil-Polluted Soil Samples in Kazakhstan.

For decades, researchers have focused on containing terrestrial oil pollution. The heterogeneity of soils, with immense microbial diversity, inspires them to transform pollutants and find cost-effective bioremediation methods. In this study, the mycoremediation potentials of five filamentous fungi isolated from polluted soils in Kazakhstan were investigated for their degradability of n-alkanes and branched-chain alkanes as sole carbon and energy sources. Dry weight estimation and gas chromatography-mass spectrometry (GC-MS) monitored the growth and the changes in the metabolic profile during degradation, respectively. Penicillium javanicum SBUG-M1741 and SBUG-M1742 oxidized medium-chain alkanes almost completely through mono- and di-terminal degradation. Pristane degradation by P. javanicum SBUG-M1741 was >95%, while its degradation with Purpureocillium lilacinum SBUG-M1751 was >90%. P. lilacinum SBUG-M1751 also exhibited the visible degradation potential of tetradecane and phytane, whereby in the transformation of phytane, both the mono- and di-terminal degradation pathways as well as α- and ß-oxidation steps could be described. Scedosporium boydii SBUG-M1749 used both mono- and di-terminal degradation pathways for n-alkanes, but with poor growth. Degradation of pristane by Fusarium oxysporum SBUG-M1747 followed the di-terminal oxidation mechanism, resulting in one dicarboxylic acid. These findings highlight the role of filamentous fungi in containing oil pollution and suggest possible degradation pathways.

PGPR-driven phytoremediation and physiobiochemical response of Miscanthus × giganteus to stress induced by the trace elements.

The effect of inoculation of Miscanthus × giganteus Greef et Deu by the plant growth promoting rhizobacteria (PGPRs) to the phytoremediation process and physio-biochemical plant's parameters was investigated in soil contaminated with the trace elements (TEs) from the Tekeli mining complex, Kazakhstan. Yeast Trichosporon sp. CA1, strains Rhizobium sp. Zn1-1, Shinella sp. Zn5-6, and Pseudomonas sp. CHA1-4, resistant to Zn and Pb, were isolated from the rhizosphere of M × g when the plant was cultivated in the same contaminated soil. Results illustrated that inoculation improved M × g adaptability to TEs toxicity by increasing the tolerance index to 2.9. The treatment enhanced the aboveground biomass yield by up to 163%, root biomass by up to 240%, chlorophyll content by up to 30%, and Chla/b ratio by up to 21%. Through M × g active growth and development, the peak activity of antioxidant enzymes was observed: activity of superoxide dismutase and glutathione reductase was induced, while the activity of catalase and ascorbate peroxidase was inhibited. Based on bioconcentration and translocation factors it was revealed that PGPRs selectively increased the uptake of TEs or stabilised them in the M × g rhizosphere. Inoculation with PGPRs increased the stabilization of Pb, V, Cr, Co, Ni, Cu, Cd, As, and Ba in the soil and plant tissues. Further research should focus on ex situ experiments using isolated PGPRs.

Moniliella spathulata, an oil-degrading yeast, which promotes growth of barley in oil-polluted soil.

The yeast strain Moniliella spathulata SBUG-Y 2180 was isolated from oil-contaminated soil at the Tengiz oil field in the Atyrau region of Kazakhstan on the basis of its unique ability to use crude oil and its components as the sole carbon and energy source. This yeast used a large number of hydrocarbons as substrates (more than 150), including n-alkanes with chain lengths ranging from C10 to C32, monomethyl- and monoethyl-substituted alkanes (C9-C23), and n-alkylcyclo alkanes with alkyl chain lengths from 3 to 24 carbon atoms as well as substituted monoaromatic and diaromatic hydrocarbons. Metabolism of this huge range of hydrocarbon substrates produced a very large number of aliphatic, alicyclic, and aromatic acids. Fifty-one of these were identified by GC/MS analyses. This is the first report of the degradation and formation of such a large number of compounds by a yeast. Inoculation of barley seeds with M. spathulata SBUG-Y 2180 had a positive effect on shoot and root development of plants grown in oil-contaminated sand, pointing toward potential applications of the yeast in bioremediation of polluted soils. KEY POINTS: • Moniliella spathulata an oil-degrading yeast • Increase of the growth of barley.

Optimization of microbial assisted phytoremediation of soils contaminated with pesticides.

580 microbial strains were isolated from the rhizosphere of the plants Cucurbita pepo L. and Xanthium strumarium grown on soil contaminated with dichlorodiphenyltrichloroethane (DDT) and its metabolites. During the cultivation, two bacterial strains were selected because of their ability to grow on media containing 0.5-5.0 mg L-1 of dichlorodiphenyldichloroethylene (DDE) as the sole carbon source. They were identified as Bacillus vallismortis and Bacillus aryabhattai. Both of these species were shown to have a high capacity for the utilization of DDE - more than 90% of which was consumed after 21 days of cultivation. Laboratory experiments were carried out then to assess the possibility of using these strains for the decontamination of organochlorine pesticides (OCPs) contaminated soils. Inoculation of C. pepo and X. strumarium with our isolates B. vallismortis and B. aryabhattai resulted in a reduction of the pollutant stress to the plants as shown by an increase both in aboveground and in root biomass. The microorganisms enhanced the uptake and phytostabilization potential of C. pepo and X. strumarium and can be applied for the treatment of DDE contaminated soils.

Bacterial endophytes of Trans-Ili Alatau region's plants as promising components of a microbial preparation for agricultural use.

In this study, 382 isolates of bacterial endophytes from tissues of plants grown in the foothills and piedmont plains of the Trans-Ili Alatau were isolated. It was found that certain strains actively produce indole-3-acetic acid (IAA) in a medium containing l-tryptophan. Among the strains studied, 26 strains (66%) showed a positive response to production of IAA. Bacteria synthesized IAA in the range of 18.6 ± 1.1 to 82.4 ± 2.3 µg/ml. IAA was synthesized more actively by Streptosporangium sp. KK1 (44.1 µg/ml), Rhodococcus sp. KK 2 (42.5 µg/ml), Streptomyces tendae KK3 (44.9 µg/ml) strains. The most active auxin's producer is a Jeotgalicoccus halotolerans BAK1 strain, whose total level of IAA production reached 82.4 µg/ml. Such strains as Streptomyces griseorubiginosis KK4, Streptomyces sp. KK5 and Jeotgalicoccus halotolerans BAK1 were found to have a significant stimulating effect which was reflected in the increase of the length of the roots of soybean and barley. As a result, 8 promising strains with fungicidal, growth-stimulating, phosphorus dissolving and enzymatic activities were selected for the further development of highly microbiological preparations for crop research.

The effect of application of micromycetes on plant growth, as well as soybean and barley yields.

The possibility of application of micromycetes (strains Penicillium bilaiae Pb14, Aureobasidium pullulans YA05 and Rhodotorula mucilaginosa YR07) to increase yields of soybean (Glycine max cv Almaty) and barley (Hordeum vulgare cv Arna) was estimated. It was shown that the most positive effect on germination energy and seed germination after seed treatment with liquid culture, supernatant and filtrate, is achieved at 1:5 dilution. In studying the influence of cell-associated and extracellular biologically active compounds of micromycetes (liquid culture and supernatant) on biometric parameters of seedlings, the maximum stimulating effect was observed in the variants with liquid culture. These strains of micromycetes were used as a bases for various compositions of preparations - application of each strain separately and application of micromycetes mixes. In microfield experiments, the increase of soybean yield ranged from 4.5 to 9.4 quintal/ha, barley - from 2.9 to 5.9 quintal/ha. A significant increase in various parameters of structure of the yield was shown in all experimental variants when compared to the control. It was found that an increase in soybean and barley yields and yield components was higher in the variant with a mix of micromycetes when compared to the separate application of each strain. The most efficient mixture was based on the mix of fungal strains (culture filtrate of P. bilaiae Pb14 diluted 1:5 + liquid cultures of A. pullulans YA05 and Rh. mucilaginosa YR07 in a 1:5 dilution).

Plant growth-promoting and antifungal activity of yeasts from dark chestnut soil.

538 yeast strains were isolated from dark chestnut soil collected from under the plants of the legume family (Fabaceae). The greatest number of microorganisms is found at soil depth 10-20 cm. Among the 538 strains of yeast 77 (14.3%) strains demonstrated the ability to synthesize IAA. 15 strains were attributed to high IAA-producing yeasts (above 10 µg/ml). The most active strains were YA05 with 51.7 ± 2.1 µg/ml of IAA and YR07 with 45.3 ± 1.5 µg/ml. In the study of effect of incubation time on IAA production the maximum accumulation of IAA coincided with maximum rates of biomass: at 120 h for YR07 and at 144 h for strain YA05. IAA production increased when medium was supplemented with the L-tryptophan. 400 µg/ml of L-tryptophan showed maximum IAA production. 10 strains demonstrated the ability to inhibit the growth and development of phytopathogenic fungi. YA05 and YR07 strains formed the largest zones of inhibition compared to the other strains--from 21.6 ± 0.3 to 30.6 ± 0.5 mm. Maximum zone of inhibition was observed for YA05 against Phytophtora infestans and YR07 strains against Fusarium graminearum. YA05 and YR07 strains were identified as Aureobasidium pullulans YA05 (GenBank accession No JF160955) and Rhodotorula mucilaginosa YR07 (GenBank accession No JF160956).

Enrichment of aliphatic, alicyclic and aromatic acids by oil-degrading bacteria isolated from the rhizosphere of plants growing in oil-contaminated soil from Kazakhstan.

Three microbial strains were isolated from the rhizosphere of alfalfa (Medicago sativa), grass mixture (Festuca rubra, 75 %; Lolium perenne, 20 %; Poa pratensis, 10 %), and rape (Brassica napus) on the basis of their high capacity to use crude oil as the sole carbon and energy source. These isolates used an unusually wide spectrum of hydrocarbons as substrates (more than 80), including n-alkanes with chain lengths ranging from C12 to C32, monomethyl- and monoethyl-substituted alkanes (C12-C23), n-alkylcyclo alkanes with alkyl chain lengths from 4 to 18 carbon atoms, as well as substituted monoaromatic and diaromatic hydrocarbons. These three strains were identified as Gordonia rubripertincta and Rhodococcus sp. SBUG 1968. During their transformation of this wide range of hydrocarbon substrates, a very large number of aliphatic, alicyclic, and aromatic acids was detected, 44 of them were identified by GC/MS analyses, and 4 of them are described as metabolites for the first time. Inoculation of plant seeds with these highly potent bacteria had a beneficial effect on shoot and root development of plants which were grown on oil-contaminated sand.