Isolation of bacteria from Grifola frondosa cultivation on wood logs to find mycelial growth-promoting bacteria.

This study aimed to isolate bacteria that coexist with the edible mushroom Grifola frondosa when it is cultured on wood, and to determine their interactions; in turn, the aim was to find bacteria that stimulate mycelial growth so as to decrease the time required for spawn preparation on potato dextrose agar (PDA). Some Pseudomonas, Dyella, Bacillus, and Priestia spp. isolated from the cultivation surroundings of G. frondosa had a positive effect on mycelial growth of the fungus in PDA. However, some isolated bacteria had a severe negative effect on the mycelial growth, especially Burkholderia spp. Thus, both mycelial-promoting bacteria and potentially pathogenic bacteria coexist with G. frondosa in cultivation. Enzyme activity assays indicated that some wood-degrading bacteria inhabit the cultivation surroundings of G. frondosa, and these bacteria probably help the fungus to degrade wood (especially cellulose).

Biodecolorization and biotransformation of methylene blue using mixed cultures of brown-rot fungus Daedalea dickinsii and filamentous fungus Aspergillus oryzae: identification of metabolites and degradation pathway.

This study aimed to examine biodecolorization and biotransformation of methylene blue (MB) using mixed cultures of brown-rot fungus Daedalea dickinsii and filamentous fungus Aspergillus oryzae. In addition, the ratio of D. dickinsii and A. oryzae in mixed cultures was 1 : 1, and the sample was incubated at 30 °C for 7 days in liquid medium potato dextrose broth (PDB). The results showed that the sample had the ability to remove and transform 95.24 mg L-1 MB. In this study, mixed cultures had the highest removal percentage of 64.77%, while values of 5.94% and 36.82% were obtained for single cultures of D. dickinsii and A. oryzae, respectively. LC-TOF/MS analysis results showed that peak intensity of MB compound (m/z 284) in each treatment chromatogram decreased compared to the control. The metabolites of decolorization by D. dickinsii were C15H16N3S, C16H19N3SO, and C16H21N3SO, while C31H48N3S+ was obtained using A. oryzae. For mixed cultures, the metabolites obtained included C26H37N2O3S, C9H8N2O3S, C28H38NO2S, and C27H27N5S2. Based on the results, mixed cultures of D. dickinsii and A. oryzae had a high MB decolorization and could be used in the textile industry.

Culture Conditions for Mycelial Growth and Anti-Cancer Properties of Termitomyces.

Termitomyces sp. that grow in symbiosis with fungus-farming Termites have medicinal properties. However, they are rare in nature, and their artificial culture is challenging. The expression of AXL receptor tyrosine kinase and immune checkpoint molecules favor the growth of cancer cells. The study evaluated the optimal conditions for the artificial culture of Termitomyces and their inhibitory activity on AXL and immune checkpoint molecules in lung adenocarcinoma and melanoma cell lines. The culture of 45 strains of Termitomyces was compared. Five strains with marked growth rates were selected. Four of the selected strains form a single cluster by sequence analysis. The mycelium of 4 selected strains produces more fungal mass in potato dextrose broth than in a mixed media. The bark was the most appropriate solid substrate for Termitomyces mycelia culture. The mycelium of all five selected strains showed a higher growth rate under normal CO2 conditions. The culture broth, methanol, and ethyl acetate of one selected strain (T-120) inhibited the mRNA relative expression of AXL receptor tyrosine kinase and immune checkpoint molecules in cancer cell lines. Overall, these results suggest the potential usefulness of Termitomyces extracts as a co-adjuvant therapy in malignant diseases.

Upregulation of MAP kinase HOG1 gene of white-rot fungus Phlebia sp. MG-60 inhibits the ethanol fermentation and mycelial growth.

Wood biomass conversion for fossil resource replacement could result in the sustainable production of chemicals, although lignin represents an obstacle to efficient polysaccharide use. White-rot fungus Phlebia sp. MG-60 reportedly selectively and aerobically degrades lignin in hardwood, then it begins cellulose saccharification from the delignified wood to produce ethanol. Environmental conditions might change white-rot fungi-driven biomass conversion. However, how the environmental response sensor affects ethanol fermentation in white-rot fungi remains elusive. In this study, we focused on MGHOG1, the yeast Hog1 homolog in Phlebia sp. MG-60, a presumably important player in osmoresponse. We generated MGHOG1 overexpressing (OE) transformants in Phlebia sp. MG-60, exhibiting slower mycelial growth compared with the wild-type under salinity stress. MGHOG1 overexpressing liquid cultures displayed suppressed mycelial growth and ethanol fermentation. Therefore, MGHOG1 potentially influences ethanol fermentation and mycelial growth in Phlebia sp. MG-60. This study provides novel insights into the regulation of white-rot fungi-mediated biomass conversion.

A novel gene, Le-Dd10, is involved in fruiting body formation of Lentinula edodes.

The cDNA library prepared from Lentinula edodes, Hokken 600 (H600), primordia was screened using cDNA expressed specifically in Dictyostelium discoideum prestalk as a probe. Twenty-one clones, Le-Dd1 ~ 21, were isolated from the L. edodes primordia cDNA library. Functional analysis of each gene was carried out by transformation into protoplast cells from L. edodes Mori 252 (M252) mycelia with the overexpression vector pLG-RasF1 of each gene because M252 protoplast cells were transformed with an 11-fold higher efficiency than H600 cells. Transformants with the overexpression vector of Le-Dd10 formed a fruiting body at almost the same time as H600, a positive control, although M252, a negative control, did not form a fruiting body under culture conditions. This suggested that Le-Dd10 is involved in the formation of fruiting bodies. Single-strand conformation polymorphism analysis revealed that Le-Dd10 is located on No. 4 linkage group of L. edodes. The properties of Le-Dd10 products were investigated by Western blotting analysis using polyclonal antibodies against GST:Le-Dd10 fusion proteins. As a result, 56-kDa, 27-kDa, and 14-kDa protein bands appeared in primordial and fruiting body stages, although the expected molecular weight of the Le-Dd10 product was 50 kDa.

Deposition patterns of feruloylarabinoxylan during cell wall formation in moso bamboo.

MAIN CONCLUSION: The feruloylarabinoxylan deposition was initiated at the formation of the secondary cell wall, especially S2 layer in moso bamboo, which may affect crosslinking between cell wall components and plant growth. Hemicelluloses, major components of plant cell walls that are hydrogen bonded to cellulose and covalently bound to lignin, are crucial determinants of cell wall properties. Especially in commelinid monocotyledons, arabinoxylan is often esterified with ferulic acid, which is essential to crosslinking with cell wall components. However, the deposition patterns and localization of ferulic acid during cell wall formation remain unclear. In this study, developing moso bamboo (Phyllostachys pubescens) culms were used to elucidate deposition patterns of hemicelluloses including feruloylarabinoxylan. Ferulic acid content peaked with cessation of elongation growth, and thereafter decreased and remained stable as culm development proceeded. During primary cell wall (PCW) formation, xyloglucan and (1,3;1,4)-ß-glucan signals were detected in all tissues. Along with culm development, arabinoxylan and feruloylarabinoxylan signals were sequentially observed in the protoxylem, vascular fibers and metaxylem, and parenchyma. Feruloylarabinoxylan signals were observed slightly later than arabinoxylan signals. Arabinoxylan signals were observed throughout the compound middle lamella and secondary cell wall (SCW), whereas the feruloylarabinoxylan signal was localized to the S2 layer of the SCW. These results indicate that the biosynthesis of hemicelluloses is regulated in accordance with cell wall layers. Feruloylarabinoxylan deposition may be initiated at the formation of SCW, especially S2 layer formation. Ferulic acid-mediated linkages of arabinoxylan-arabinoxylan and arabinoxylan-lignin would arise during SCW formation with the cessation of elongation growth.

Performance of stacked microbial fuel cells with barley-shochu waste.

The treatment of barley-shochu waste combined with electricity generation was examined using stacked microbial fuel cells (SMFCs). The maximum chemical oxygen demand (CODCr) removal efficiency and maximum power density were achieved at 36.7 ± 1.1% and 4.3 ± 0.2 W m⁻³ (15.7 ± 0.9 mW m-2). The acetic acid concentration in effluent increased, whereas the citric acid, ethanol and sugar concentrations decreased during the operation. Microbial community analysis of the anode cell suspension and raw barley-shochu waste revealed that Clostridiaceae, Acetobacteraceae, and Enterobacteriaceae became predominant after the operation, implying that microorganisms belonging to these families might be involved in organic waste decomposition and electricity generation in the SMFCs.

Development of an 111In-Labeled Glucagon-Like Peptide-1 Receptor-Targeting Exendin-4 Derivative that Exhibits Reduced Renal Uptake.

Insulinomas are neuroendocrine tumors that are mainly found in the pancreas. Surgical resection is currently the first-line treatment for insulinomas; thus, it is vital to preoperatively determine their locations. The marked expression of the glucagon-like peptide-1 receptor (GLP-1R) is seen in pancreatic ß-cells and almost all insulinomas. Radiolabeled derivatives of exendin-4, a GLP-1R agonist, have been used with nuclear medicine imaging techniques for the in vivo detection of the GLP-1R; however, their marked renal accumulation can hinder the imaging of pancreatic tail lesions. To develop a GLP-1R imaging probe that exhibits reduced renal accumulation, we designed and synthesized a straight-chain GLP-1R-targeting radioligand, [111In]In-E4DA1, which consisted of exendin-4, DOTADG (a chelator), and an (iodophenyl)butyric acid derivative (an albumin binder [ALB]). We performed preclinical evaluations of [111In]In-E4DA1 to investigate its utility as a GLP-1R imaging probe. [111In]In-E4DA1 and [111In]In-E4D (a control compound lacking the ALB moiety) were prepared by reacting the corresponding precursors with [111In]InCl3 in buffer. Cell-binding and human serum albumin (HSA)-binding assays were performed to assess the in vitro affinity of the molecules for INS-1 (GLP-1R-positive) cells and albumin, respectively. A biodistribution assay and single-photon emission computed tomography imaging were carried out using INS-1 tumor-bearing mice. In the cell-binding assay, [111In]In-E4DA1 and [111In]In-E4D exhibited in vitro binding to INS-1 cells. In the HSA-binding assay, [111In]In-E4DA1 bound to HSA, while [111In]In-E4D showed little HSA binding. The in vivo experiments involving INS-1 tumor-bearing mice revealed that the introduction of an ALB moiety into the DOTADG-based exendin-4 derivative markedly increased the molecule's tumor accumulation while decreasing its renal accumulation. In addition, [111In]In-E4DA1 exhibited greater tumor accumulation than renal accumulation, whereas previously reported radiolabeled exendin-4 derivatives demonstrated much higher accumulation in the kidneys than in tumors. These results indicate that [111In]In-E4DA1 may be a useful GLP-1R imaging probe, as it demonstrates only slight renal accumulation.

Selective Homologous Expression of Recombinant Manganese Peroxidase Isozyme of Salt-Tolerant White-Rot Fungus Phlebia sp. MG-60, and Its Salt-Tolerance and Thermostability.

Phlebia sp. MG-60 is the salt-tolerant, white-rot fungus which was isolated from a mangrove forest. This fungus expresses three kinds of manganese peroxidase (MGMnP) isozymes, MGMnP1, MGMnP2 and MGMnP3 in low nitrogen medium (LNM) or LNM containing NaCl. To date, there have been no reports on the biochemical salt-tolerance of these MnP isozymes due to the difficulty of purification. In present study, we established forced expression transformants of these three types of MnP isozymes. In addition, the fact that this fungus hardly produces native MnP in a high-nitrogen medium (HNM) was used to perform isozyme-selective expression and simple purification in HNM. The resulting MGMnPs showed high tolerance for NaCl compared with the MnP of Phanerochaete chrysosporium. It was worth noting that high concentration of NaCl (over 200 mM to 1200 mM) can enhance the activity of MGMnP1. Additionally, MGMnP1 showed relatively high thermo tolerance compared with other isozymes. MGMnPs may have evolved to adapt to chloride-rich environments, mangrove forest.

Active Transport of Lignin Precursors into Membrane Vesicles from Lignifying Tissues of Bamboo.

Lignin is the second most abundant natural polymer on Earth and is a major cell wall component in vascular plants. Lignin biosynthesis has three stages: biosynthesis, transport, and polymerization of its precursors. However, there is limited knowledge on lignin precursor transport, especially in monocots. In the present study, we aimed to elucidate the transport mode of lignin monomers in the lignifying tissues of bamboo (Phyllostachys pubescens). The growth manners and lignification processes of bamboo shoots were elucidated, which enabled us to obtain the lignifying tissues reproducibly. Microsomal membrane fractions were prepared from tissues undergoing vigorous lignification to analyze the transport activities of lignin precursors in order to show the ATP-dependent transport of coniferin and p-glucocoumaryl alcohol. The transport activities for both precursors depend on vacuolar type H+-ATPase and a H+ gradient across the membrane, suggesting that the electrochemical potential is the driving force of the transport of both substrates. These findings are similar to the transport properties of these lignin precursors in the differentiating xylem of poplar and Japanese cypress. Our findings suggest that transport of coniferin and p-glucocoumaryl alcohol is mediated by secondary active transporters energized partly by the vacuolar type H+-ATPase, which is common in lignifying tissues. The loading of these lignin precursors into endomembrane compartments may contribute to lignification in vascular plants.

Onion (Allium cepa L.)-Derived Nanoparticles Inhibited LPS-Induced Nitrate Production, However, Their Intracellular Incorporation by Endocytosis Was Not Involved in This Effect on RAW264 Cells.

The aim of this study was to evaluate the involvement of nanoparticles prepared from Allium cepa L. as anti-inflammatory agents. In the present study, we identified nanoparticles from Allium cepa L. using the ultracentrifugation exosome purification method. The nanoparticles were referred to as 17,000× g and 200,000× g precipitates, and they contained quercetins, proteins, lipids, and small-sized RNA. The nanoparticles inhibited nitric oxide production from lipopolysaccharide (LPS)-stimulated RAW264 cells without cytotoxic properties. Cellular incorporation was confirmed by laser microscopic observation after PKH26 staining. The inhibition of caveolae-dependent endocytosis and macropinocytosis significantly prevented the incorporation of the nanoparticles but had no effect on the inhibition of nitric oxide in RAW264 cells. Collectively, the identified nanoparticles were capable of inhibiting the LPS response via extracellular mechanisms. Taken together, the way of consuming Allium cepa L. without collapsing the nanoparticles is expected to provide an efficient anti-inflammatory effect.

Interaction and Effects of Bacteria Addition on Dichlorodiphenyltrichloroethane Biodegradation by Daedalea dickinsii.

The residue of organochlorine pesticides (OCPs) has been a major pollution problem in our environment. Dichlorodiphenyltrichloroethane (DDT) is one of the most common persistent OCPs that continue to pose a serious risk to human health and the environment. Some treatment methods have been developed to reduce and minimize the adverse impacts of the use of DDT, including biodegradation with brown-rot fungi (BRF). However, DDT degradation using BRF has still low degradation rate and needs a long incubation time. Therefore, the ability of BRF need to be enhanced to degrade DDT. Interaction and effect of bacteria addition on biodegradation of DDT by brown-rot fungus Daedalea dickinsii were investigated. The interaction assay between D. dickinsii with bacteria addition showed that the addition of bacterium Pseudomonas aeruginosa did not provide resistance to the growth of D. dickinsii. Meanwhile, bacterium Bacillus subtilis addition has an inhibitory effect on the growth of D. dickinsii. The addition of 10 ml (1 ml = 1.05 × 109 CFU/ml bacteria cell) of P. aeruginosa and B. subtilis was able to improve DDT biodegradation by D. dickinsii from 53.61% to 96.70% and 67.60%, respectively. The highest biodegradation capability of DDT was obtained through addition of 10 ml of P. aeruginosa into the D. dickinsii culture in which the mixed cultures produce final metabolites of 1,1-dichloro-2,2-bis(4-chlorophenyl)ethane (DDD) and 1-chloro-2,2-bis(4-chlorophenyl)ethylene (DDMU). This study indicated that the addition of P. aeruginosa can be used for optimization of DDT biodegradation by D. dickinsii.

Conservation of Xylose Fermentability in Phlebia Species and Direct Fermentation of Xylan by Selected Fungi.

In efforts to lower the cost of total conversion of lignocellulosic materials, utilization of hemicellulose must be considered. White-rot fungus Phlebia sp. MG-60 can produce ethanol directly from cellulose and has fermentation ability for glucose, cellulose, and xylose. Therefore, white-rot fungi can be considered a good candidate for consolidated bioprocessing to give bioethanol from lignocellulosic biomass, although little information is available on the direct fermentation of xylan. In the present study, some Phlebia species were selected as candidates because of their ability to ferment xylose to ethanol more efficiently than Phlebia sp. MG-60. This process indicated that the basidiomycetes that can produce ethanol from xylose are closely related genetically within the Phlebia genus. The selected Phlebia species showed higher ethanol productivity from corn core and beechwood xylans than Phlebia sp. MG-60. The ethanol yields from corn core xylan in culture with Phlebia acerina HHB11146, Phlebia ludoviciana HHB9640, and Phlebia subochracea HHB8494 were 46.2%, 46.7%, and 39.7% of theoretical maximum, and those from beechwood xylan were 19.09%, 17.7%, and 21.4% of the theoretical maximum, respectively.

Synergistic interaction of a consortium of the brown-rot fungus Fomitopsis pinicola and the bacterium Ralstonia pickettii for DDT biodegradation.

1,1,1-Trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) is a toxic and recalcitrant pesticide that has been greatly used to eradicate malaria mosquitos since the 1940s. However, the US Environmental Protection Agency banned and classified DDT as priority pollutants due to its negative impact on wildlife and human health. Considering its negative effects, it is necessary to develop effective methods of DDT degradation. A synergistic interaction of a consortium consisting of the brown-rot fungus Fomitopsis pinicola and the bacterium Ralstonia pickettii was adopted to degrade DDT. For the microbial consortia, F. pinicola was mixed with R. pickettii at 1, 3, 5, 7 and 10 ml (1 ml ≈ 1.44 × 1013 CFU) in a potato dextrose broth (PDB) medium to degrade DDT throughout the seven days incubation period. The degradation of DDT by only the fungus F. pinicola was roughly 42%, while by only R. pickettii was 31%. The addition of 3 ml of R. pickettii into F. pinicola culture presented appropriate optimization for efficient DDT degradation at roughly 61%. The DDT transformation pathway by co-inoculation of F. pinicola and R. pickettii showed that DDT was converted to 1,1-dichloro-2,2-bis(4-chlorophenyl) ethane (DDD), further transformed to 1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene (DDE), and then ultimately transformed to 1-chloro-2,2-bis(4-chlorophenyl) ethylene (DDMU). These metabolites are less toxic than DDT. This research showed that R. picketti synergistically interacts with F. pinicola by enhancing DDT degradation.

Butanol production from cellulosic material by anaerobic co-culture of white-rot fungus Phlebia and bacterium Clostridium in consolidated bioprocessing.

Butanol production from lignocelluloses is desirable. Unfortunately, the known wild-types of butanol fermenting Clostridium bacteria are not capable of delignification and saccharification. Here we analyzed butanol production from cellulosic material using anaerobic co-culture of C. saccharoperbutylacetonicum with the white-rot fungus Phlebia sp. MG-60-P2. In consolidated bioprocessing, the co-culture synergistically produced butanol and enhanced saccharification. Knockout of the pyruvate decarboxylase gene from MG-60-P2 to produce transformant line KO77 led to inhibition of ethanol fermentation and high accumulation of saccharified cellobiose and glucose from cellulose. In co-culture of KO77 with C. saccharoperbutylacetonicum, enhanced butanol production was observed (3.2 g/L, compared with 2.5 g/L in co-culture of MG-60-P2 and C. saccharoperbutylacetonicum). We believe this is the first application of co-culture between white-rot fungus and Clostridium to produce butanol from cellulose; butanol production from lignocellulose by co-culture of C. saccharoperbutylacetonicum with Phlebia sp. MG-60-P2 and its transformant should be pursued.

Growth management of white-rot fungus Phlebia brevispora improved degradation of high-molecular-weight polycyclic aromatic hydrocarbons.

The effect of co-culturing white-rot fungus Phlebia brevispora with growth-promoting bacterial strains Enterobacter sp. TN3W-14 and Pseudomonas sp. TN3W-8 on the degradation of polycyclic aromatic hydrocarbons (PAHs) was evaluated in liquid culture. In axenic cultures, P. brevispora strains TN3F and TMIC33929 showed high degradation of phenanthrene (> 98%) within 15 days, and degraded 65% and 63% of pyrene, and 12% and 8% of benzo(a)pyrene, respectively. This low level of degradation ability toward benzo(a)pyrene was improved significantly by co-culturing the fungi with a mixture of bacterial strains TN3W-8 and TN3W-14 (mixed bacterial co-culture; MBC). Within 15 days, P. brevispora TN3F with MBC achieved about 86% pyrene and 53% benzo(a)pyrene degradation; P. brevispora TMIC33929 with MBC showed 92% pyrene and 72% benzo(a)pyrene degradation. The MBC alone degraded little PAH, as its growth was inhibited by PAH; however, its co-culture with P. brevispora improved mycelial growth of the fungus, which led to improved degradation of the PAHs. A possible dihydrodiol metabolite of pyrene from fungal cultures suggests that hydroxylation was the first step in the degradation of pyrene by P. brevispora.

Ralstonia pickettii Enhance the DDT Biodegradation by Pleurotus eryngii.

DDT is a hydrophobic organic pollutant, which can be bio-accumulated in nature and have adverse consequences on the physical condition of humans and animals. This study investigated the relationship between the white-rot fungus Pleurotus eryngii and biosurfactantproducing bacterium Ralstonia pickettii associated with the degradation of DDT. The effects of R. pickettii on fungal development were examined using in vitro confrontation assay on a potato dextrose agar (PDA) medium. R. pickettii culture was added to the P. eryngii culture at 1, 3, 5, 7, and 10 ml (1 ml ≈ 1.44 × 1013 CFU). After 7 d incubation, about 43% of the initial DDT (12.5 µM) was degraded by the P. eryngii culture only. The augmentation of 7 ml of R. pickettii culture revealed a more highly optimized synergism with DDT degradation being approximately 78% and the ratio of optimization 1.06. According to the confrontational assay, R. pickettii promoted the growth of P. eryngii towards the bacterial colony, with no direct contact between the bacterial cells and mycelium (0.71 cm/day). DDD (1,1-dichloro-2,2-bis(4- chlorophenyl) ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), and DDMU (1- chloro-2,2-bis(4-chlorophenyl) ethylene) were identified as metabolic products, indicating that the R. pickettii could enhance the DDT biodegradation by P. eryngii.

Proton Gradient-Dependent Transport of p-Glucocoumaryl Alcohol in Differentiating Xylem of Woody Plants.

Lignin is a cell wall component of vascular plants crucial for survival in terrestrial environments. While p-hydroxyphenyl lignin is minor, it is considered to be localised in the outermost part of the cell wall providing strong adhesion between cells, which determines cell shape. Transport of the lignin precursor from the cytosol to the cell wall is critical to regulate temporal and spatial lignin deposition; however, little information on the transport step is available. Here, we report transport activity of p-glucocoumaryl alcohol, a precursor of p-hydroxyphenyl lignin, in a broad-leaved tree (hybrid poplar, Populus sieboldii × P. grandidentata) and a coniferous tree (Japanese cypress, Chamaecyparis obtusa). Membrane vesicles of both trees were prepared from differentiating xylem with vigorous lignification and used for transport assays. Several inhibition assays indicated that not ABC transporters but the proton gradient and V-ATPase are involved in p-glucocoumaryl alcohol transport depending on ATP. These results support the hypothesis that p-glucocoumaryl alcohol is loaded into the secretory vesicles and delivered to the cell wall by exocytosis. Furthermore, this transport mechanism was common in both poplar and Japanese cypress, strongly suggesting that p-glucocoumaryl alcohol transport in the differentiating xylem is conserved within woody plants.

Characteristics of White-rot Fungus Phlebia brevispora TMIC33929 and Its Growth-Promoting Bacterium Enterobacter sp. TN3W-14 in the Decolorization of Dye-Contaminated Water.

The dye decolorization potential of the white-rot fungus Phlebia brevispora TMIC33929 when grown alone or in coculture with its growth-promoting bacterium Enterobacter sp. TN3W-14 was evaluated in low nitrogen liquid medium at different pHs. Axenic fungus removed a similar amount of Congo red and crystal violet at pH 4.5 and 7.0, respectively. The bacterium alone achieved only slightly better decolorization of crystal violet than the fungus at pH 9.0. Compared with axenic fungus, cocultures provided no increased crystal violet removal but achieved higher removal of crystal violet in mixed dye at all pHs, and the best-mixed dye decolorization at pH 9.0. Unlike bacterial growth on dyes, growth of fungal mycelia was not inhibited by the dyes at all pH but the cocultures gave comparably higher mycelial growth.

Indirect Bacterial Effect Enhanced Less Recovery of Neonicotinoids by Improved Activities of White-Rot Fungus Phlebia brevispora.

Bacterial strains that improve mycelial morphology and growth of white-rot fungi in liquid medium could enhance the impact of white-rot fungi towards lesser recovery of neonicotinoids when cocultured. This was demonstrated by the recovery of clothianidin and acetamiprid from cocultures of the white-rot fungus Phlebia brevispora strains with two mycelial-growth-promoting bacteria, Enterobacter sp. TN3W-14 and Pseudomonas sp. TN3W-8. Clothianidin recovery from cocultures of white-rot fungi and bacteria was over 40% lower than that from axenic microbial cultures and mixed-bacterial cultures. About 20% less acetamiprid was equally recovered from both TMIC33929+TN3W-14 cocultures and mixedbacterial cultures than from axenic fungal and bacterial cultures.