Application of Fungus Enzymes in Spent Mushroom Composts from Edible Mushroom Cultivation for Phthalate Removal.

Spent mushroom composts (SMCs) are waste products of mushroom cultivation. The handling of large amounts of SMCs has become an important environmental issue. Phthalates are plasticizers which are widely distributed in the environment and urban wastewater, and cannot be effectively removed by conventional wastewater treatment methods. In this study, SMCs are tested for their ability to remove phthalates, including benzyl butyl phthalate (BBP), di-n-butyl phthalate (DBP), and diethyl phthalate (DEP). Batch experiments reveal that BBP, DBP, and DEP can be degraded by the SMC enzyme extracts of four edible mushrooms: Pleurotus eryngii, Pleurotus djamor, Pleurotus ostreatus, and Auricularia polytricha. Potential fungus enzymes associated with BBP, DBP, and DEP degradation in SMCs (i.e., esterases, oxygenases, and oxidases/dehydrogenases) are uncovered by metaproteomic analysis using mass spectrometry. Bioreactor experiments indicate that the direct application of SMCs can remove BBP, DBP, and DEP from wastewater, through adsorption and biodegradation. The results of this study extend the application of white-rot fungi without laccases (e.g., Auricularia sp.) for the removal of organic pollutants which are not degraded by laccases. The application of SMCs for phthalate removal can be developed into a mycoremediation-based green and sustainable technology.

Effects of sulfamethoxazole and sulfamethoxazole-degrading bacteria on water quality and microbial communities in milkfish ponds.

Intensive farming practices are typically used for aquaculture. To prevent disease outbreaks, antibiotics are often used to reduce pathogenic bacteria in aquaculture animals. However, the effects of antibiotics on water quality and microbial communities in euryhaline fish culture ponds are largely unknown. The aim of this study was to investigate the interactions between sulfamethoxazole (SMX), water quality and microbial communities in milkfish (Chanos chanos) culture ponds. The results of small-scale milkfish pond experiments indicated that the addition of SMX decreased the abundance of ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB) and photosynthetic bacteria. Consequently, the levels of ammonia and total phosphorus in the fish pond water increased, causing algal and cyanobacterial blooms to occur. In contrast, the addition of the SMX-degrading bacterial strains A12 and L effectively degraded SMX and reduced the levels of ammonia and total phosphorus in fish pond water. Furthermore, the abundances of AOB, NOB and photosynthetic bacteria were restored, and algal and cyanobacterial blooms were inhibited. This study demonstrate the influences of SMX on water quality and microbial community composition in milkfish culture ponds. Moreover, the use of the bacterial strains A12 and L as dual function (bioaugmentation and water quality maintenance) beneficial bacteria was shown to provide an effective approach for the bioremediation of SMX-contaminated euryhaline milkfish culture ponds.

Anaerobic degradation of sulfamethoxazole by mixed cultures from swine and sewage sludge.

The objective of this study was to evaluate the anaerobic degradation of sulfamethoxazole (SMX) and the associated bacterial community changes in swine and sewage sludges. The degradation rate of SMX was higher in swine sludge than in sewage sludge. The addition of lactate, citrate, and sucrose had significant effects on SMX degradation, and sucrose addition yielded a higher SMX degradation rate than the other additives. At concentrations of 0.1-10 g/l sucrose, the SMX degradation rates increased in the sludge. The bacterial genera from swine sludge with sucrose exhibited the highest SMX degrading efficiency. Seventeen bacterial genera were found to be the major bacterial community members involved in SMX degradation in the sludge.

Anaerobic degradation of sulfamethoxazole in mangrove sediments.

The effects of sucrose and electron acceptors on the anaerobic degradation of sulfamethoxazole (SMX) in mangrove sediments were investigated in this study. Among three sulfonamides, sulfamethoxazole, sulfadimethoxine and sulfamethazine, only SMX could be completely degraded in mangrove sediments. Degradation of SMX was enhanced by the addition of sucrose to the sediments. The degradation rates of SMX were increased in bioreactor experiments with sucrose. The addition of electron acceptors (sodium hydrogen carbonate, sodium sulfate, and sodium nitrate) could further enhance SMX degradation. The order of anaerobic SMX degradation rates under three different conditions was as follows: sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. Methanolobus, Desulfuromonas, and Thauera were found in the highest proportions among methanogens, sulfate-reducing bacteria and denitrifying bacteria, respectively. Achromobacter, Brevundimonas, Delftia, Idiomarina, Pseudomonas, and Rhodopirellula were the major bacterial communities responsible for SMX degradation in the sediment. Overall, 16 bacterial and archaeal genera were identified as the core microbial community facilitating anaerobic SMX degradation for all methanogenic, sulfate-reducing and nitrate-reducing conditions. The results of this study provide feasible methods for the removal of SMX from mangrove sediments.

Removal of emerging contaminants using spent mushroom compost.

Acetaminophen and sulfonamides are emerging contaminants. Conventional wastewater treatment systems fail to degrade these compounds properly. Mycoremediation, is a form of novel bioremediation that uses extracellular enzymes of white-rot fungi to degrade pollutants in the environment. In this study, spent mushroom compost (SMC), which contains fungal extracellular enzymes, was tested for acetaminophen and sulfonamides removal. Among the SMCs of nine mushrooms tested in batch experiments, the SMC of Pleurotus eryngii exhibited the highest removal rate for acetaminophen and sulfonamides. Several fungal extracellular enzymes that might be involved in removal of acetaminophen and sulfonamides were identified by metaproteomic analysis. The bacterial classes, Betaproteobacteria and Alphaproteobacteria, were revealed by metagenomic analysis and may be assisting with acetaminophen and sulfonamide removal, respectively, in the SMC of Pleurotus eryngii. Bioreactor experiments were used to simulate the capability of Pleurotus eryngii SMC for the removal of acetaminophen and sulfonamides from wastewater. The results of this study provide a feasible solution for acetaminophen and sulfonamide removal from wastewater using the SMC of Pleurotus eryngii.

Fungi extracellular enzyme-containing microcapsules enhance degradation of sulfonamide antibiotics in mangrove sediments.

Mangroves represent a special coastal vegetation along the coastlines of tropical and subtropical regions. Sulfonamide antibiotics (SAs) are the most commonly used antibiotics. The application of white-rot fungi extracellular enzyme-containing microcapsules (MC) for aerobic degradation of SAs in mangrove sediments was investigated in this study. Degradation of three SAs, sulfamethoxazole (SMX), sulfadimethoxine (SDM), and sulfamethazine (SMZ), was enhanced by adding MC to the sediments. The order of SA degradation in batch experiments was SMX > SDM > SMZ. Bioreactor experiments revealed that SA removal rates were higher with than without MC. The enhanced SA removal rates with MC persisted with three re-additions of SAs. Thirteen bacteria genera (Achromobacter, Acinetobacter, Alcaligenes, Aquamicrobium, Arthrobacter, Brevundimonas, Flavobacterium, Methylobacterium, Microbacterium, Oligotropha, Paracoccus, Pseudomonas, and Rhodococcus) were identified to be associated with SA degradation in mangrove sediments by combination of next-generation sequencing, bacterial strain isolation, and literature search results. Results of this study suggest that MC could be used for SA removal in mangrove sediments.

Microbial communities associated with anaerobic degradation of polybrominated diphenyl ethers in river sediment.

BACKGROUND/PURPOSE: Polybrominated diphenyl ethers (PBDEs) are extensively used as a class of flame retardants and have become ubiquitous environmental pollutants. We aimed to uncover the changes in microbial community with PBDE anaerobic degradation with and without zero-valent iron in sediment from the Erren River, considered one of the most heavily contaminated rivers in Taiwan. METHODS: PBDE anaerobic degradation in sediment was analyzed by gas chromatography with an electron capture detector. Microbial community composition was analyzed by a pyrosequencing-based metagenomic approach. RESULTS: The anaerobic degradation rate of BDE-209 was higher than BDE-28 in sediment; the addition of zero-valent iron enhanced the degradation rates of both. In total, 19 known bacterial genera (4 major genera: Clostridium, Lysinibacillus, Rummeliibacillus, and Brevundimonas) were considered PBDE degradation-associated bacteria (sequence frequency negatively correlated with PBDE remaining percentage) as were four known archaea genera (Methanobacterium, Methanosarcina, Methanocorpusculum, and Halalkalicoccus; sequence frequency positively correlated with PBDE remaining percentage). CONCLUSION: The composition of bacteria and that of archaea affected the anaerobic degradation of BDE-28 and BDE-209. The addition of zero-valent iron further decreased the archaea content to undetectable levels.

Bacterial communities associated with anaerobic debromination of decabromodiphenyl ether from mangrove sediment.

This study evaluated decabromodiphenyl ether (BDE-209) anaerobic debromination and bacterial community changes in mangrove sediment. BDE-209 debromination rates were enhanced with zerovalent iron compared to without zerovalent iron in the sediment. BDE-209 debromination rates in microcosms constructed with sediments collected in autumn were higher than in microcosms constructed with sediments collected in spring and were higher at the Bali sampling site than the Guandu sampling site. The intermediate products resulting from the reductive debromination of BDE-209 in sediment were nona-BDE (BDE-206, BDE-207), octa-BDEs (BDE-196, BDE-197), hepta-BDEs (BDE-183, BDE-184, BDE-191), hexa-BDEs (BDE-137, BDE-138, BDE-154, BDE-157), penta-BDEs (BDE-85, BDE-99, BDE-100, BDE-126), tetra-BDEs (BDE-47, BDE-49, BDE-66, BDE-77), tri-BDEs (BDE-17, BDE-28), and di-BDEs (BDE-15). Fifty bacterial genera associated with BDE-209 debromination were identified. Overall, 12 of the 50 bacterial genera were reported to be involved in dehalogenation of aromatic compounds. These bacteria have high potential to be BDE-209 debromination bacteria. Different combinations of bacterial community composition exhibit different abilities for BDE-209 anaerobic debromination.

Bacterial communities associated with sulfonamide antibiotics degradation in sludge-amended soil.

This study investigated the degradation of sulfonamide antibiotics (SAs) and microbial community changes in sludge-amended soil. In batch experiments, SA degradation was enhanced by addition of spent mushroom compost (SMC), SMC extract, and extract-containing microcapsule, with SMC showing higher SA degradation rate than the other additives in soil-sludge mixtures. In bioreactor experiments, the degradation of SAs in soil-sludge mixtures was in the order of sulfamethoxazole > sulfadimethoxine > sulfamethazine during four times of SA addition. SA removal was higher in soil-sludge mixtures than in soil alone. The bacterial composition differed in soil-sludge mixtures with and without SMC. In total, 44 differentially distributed bacterial genera were identified from different experimental settings and stages. Four bacterial genera, Acinetobacter, Alcaligenes, Brevundimonas, and Pseudomonas, were previously found involved in SA degradation, and 20 of the 44 bacterial genera were previously found in aromatic hydrocarbon degradation. Therefore, these bacteria have high potential to be SA degradation bacteria in this study.

Biodegradation of sulfonamide antibiotics in sludge.

Sulfonamide antibiotics are widely used in human and veterinary medicine. This study assessed the degradation of three sulfonamides (100 mg kg(-1) each of sulfamethoxazole, sulfadimethoxine and sulfamethazine) and changes in the microbial communities of sewage sludge. Sulfamethoxazole degradation was enhanced by spent mushroom compost (SMC), SMC extract, and extract-containing microcapsules in the sludge. The degradation of sulfonamides in sludge and SMC mixtures occurred in the order of sulfamethoxazole > sulfadimethoxine > sulfamethazine. Bioreactor experiments revealed that the sulfonamides removal rates in sludge with SMC were greater than those in sludge alone. The sulfonamides removal rates were enhanced by the addition of SMC for six time additions. The sulfonamides concentrations were 200 and 500 mg kg(-1) for the first to third additions and the fourth to sixth additions, respectively. With the high correlations between TOC and the proportions of sulfonamides remaining in sludge, sulfonamides may be mineralized to a greater extent with SMC in sludge than in sludge alone. Four bacterial genera were identified from the different settings and stages of the bioreactor experiments. Acinetobacter and Pseudomonas were major bacterial communities that were responsible for sulfonamide degradation in sludge.

Biodegradation of toxic chemicals by Pleurotus eryngii in submerged fermentation and solid-state fermentation.

BACKGROUND/PURPOSE: The toxic chemicals bisphenol A (BPA), bisphenol F (BPF), nonylphenol (NP), and tetrabromobisphenol A (TBBPA) are endocrine-disrupting chemicals that have consequently drawn much concern regarding their effect on the environment. The objectives of this study were to investigate the degradation of BPA, BPF, NP, and TBBPA by enzymes from Pleurotus eryngii in submerged fermentation (SmF) and solid-state fermentation (SSF), and also to assess the removal of toxic chemicals in spent mushroom compost (SMC). METHODS: BPA and BPF were analyzed by high-performance liquid chromatography; NP and TBBPA were analyzed by gas chromatography. RESULTS: NP degradation was enhanced by adding CuSO4 (1 mM), MnSO4 (0.5 mM), gallic acid (1 mM), tartaric acid (20 mM), citric acid (20 mM), guaiacol (1 mM), or 2,2'-azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid; 1 mM), with the last yielding a higher NP degradation rate than the other additives from SmF. The optimal conditions for enzyme activity from SSF were a sawdust/wheat bran ratio of 1:4 and a moisture content of 5 mL/g. The enzyme activities were higher with sawdust/wheat bran than with sawdust/rice bran. The optimal conditions for the extraction of enzyme from SMC required using sodium acetate buffer (pH 5.0, solid/solution ratio 1:5), and extraction over 3 hours. CONCLUSION: The removal rates of toxic chemicals by P. eryngii, in descending order of magnitude, were SSF > SmF > SMC. The removal rates were BPF > BPA > NP > TBBPA.

Bacterial communities associated with aerobic degradation of polybrominated diphenyl ethers from river sediments.

Polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants and have therefore drawn much environmental concern. We aimed to compare aerobic degradation of different PBDE congeners under various treatments and reveal the bacterial community associated with PBDE degradation in sediment. Results of this study indicate that degradation rates of BDE-15 were enhanced 45.1 and 81.3 % with the addition of suspended and microencapsulated Pseudomonas sp., respectively. However, the degradation rates of BDE-28, BDE-47, BDE-99, and BDE-100 did not differ among experimental treatments. Degradation rates of PBDE congeners were in the order of BDE-15 > BDE-28 > BDE-47 > BDE-99 > BDE-100. Using a pyrosequencing-based metagenomic approach, we found that addition of various treatments altered the microbial community composition in the sediment. Twenty-four bacterial genera associated with degradation of PBDEs; six are the core bacterial genera common among PBDE degraders. The diverse bacterial composition among different PBDE congener degradation indicates different combinations of bacteria involved in degradation of different PBDE congeners.

Biodegradation of three tetracyclines in swine wastewater.

Tetracyclines (TCs), including tetracycline (TC), oxytetracycline (OTC), and chlortetracycline (CTC), are amongst the most common antibiotics used in animal husbandry. Residual amounts of these antibiotics in the environment are a concern because they contribute to selection of resistant bacteria. In this study, we investigated the biodegradation of three TCs in swine wastewater. In batch experiments, OTC and CTC were completely degraded at d 18 and 20, respectively, but TC was remained at 7.1% after 20 d incubation. The degradation rates of TCs in the wastewater were in the order of OTC > CTC > TC. Degradation of the TCs was enhanced by the addition of enzyme extract from spent mushroom compost (SMC) of Pleurotus eryngii. The degradation rates were higher with the addition of extract-containing microcapsules than suspended enzyme extract in swine wastewater. In the bioreactor experiment, the addition of extract-containing microcapsules enhanced the removal rates of the three TCs, and adding TCs twice maintained enzyme activity in the swine wastewater. Of the microorganism strains isolated from the wastewater samples, strain HL2 (identified as Xanthobacter flavus) showed the best degrading ability.

Aerobic degradation of bisphenol-A and its derivatives in river sediment.

This study investigated the aerobic degradation ofbisphenol-A (BPA) and the derivatives bisphenol-B (BPB), bisphenol-F (BPF), tetrabromobisphenol-A (TBBPA), and tetrachlorobisphenol-A (TCBPA) in river sediment. The degradation rates of BPA and BPF were enhanced by adding brij 30, brij 35, rhamnolipid, surfactin, or crude enzyme; a higher degradation rate was observed with crude enzyme than with the other additives. The degradation rates of BPA and its derivatives (BPAs) in the sediment were BPF > BPA > BPB > TCBPA > TBBPA. Different BPAs affected the changes in the microbial community in the sediment. Sediment fractions with larger particle sizes demonstrated higher degradation rates. Different sediment particle sizes affected the changes in the microbial communities. Pseudomonas sp. may be the dominant bacteria in the process of degradation of BPAs in river sediment.

Use of microcapsules with electrostatically immobilized bacterial cells or enzyme extract to remove nonylphenol in wastewater sludge.

We investigated the use of a high-voltage electrostatic system to immobilize bacterial cells or enzyme extract in alginate microcapsules for removing nonylphenol (NP) from wastewater sludge. With applied potential increased from 0 to 12kV, the gel bead diameter decreased from 950 to 250 µm. The amount of bacterial cells or enzyme extract immobilized in alginate microcapsules was greater than that in suspension, for improved tolerance to environmental loadings. Removal of NP at 2.0-20.0 mg L(-1) was greater with extract- than cell-containing microcapsules. The percentage of toxic chemicals (2.0 mg L(-1)) removed with alginate microcapsules, in descending order of magnitude, was bisphenol-F>bisphenol-A>NP>oxytetracycline>chlortetracycline>tetracycline>dibromodiphenyl ethers>tetrabromobisphenol-A>decabromodiphenyl ether.

Removal of organic toxic chemicals using the spent mushroom compost of Ganoderma lucidum.

The removal of the organic toxic chemicals di-n-butyl phthalate (DBP), di-2-ethyl hexyl phthalate (DEHP), nonylphenol (NP), and bisphenol-A (BPA) by laccase obtained from the spent mushroom compost (SMC) of the white rot fungi, Ganoderma lucidum, was investigated. The optimal conditions for the extraction of laccase from SMC required using sodium acetate buffer (pH 5.0, solid : solution ratio 1 : 5), and extraction over 3 h at 4 °C. The removal of NP was enhanced by adding CuSO(4) (1 mM), MnSO(4) (0.5 mM), tartaric acid (20 mM), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS; 1 mM), and 1-hydroxybenzotriazole (HBT; 20 mg L(-1)), with ABTS yielding a higher NP removal efficiency than the other additives. At a concentration of 2 mg L(-1), DBP, DEHP, NP, and BPA were almost entirely removed by laccase after incubation for 1 day. The removal efficiencies, in descending order of magnitude, were DBP > BPA > NP > DEHP. We believe that these findings could provide useful information for improving the efficiency of the removal of organic toxic chemicals in the environment.

Biodegradation of dibutyl phthalate and di-(2-ethylhexyl) phthalate and microbial community changes in mangrove sediment.

In this study, we investigated the microbial degradation of the phthalate esters (PAEs) dibutyl phthalate (DBP) and di-(2-ethylhexyl) phthalate (DEHP), and change in microbial communities in mangrove sediment collected from 5 sampling sites along the Tanshui River in Taiwan. Aerobic degradation half-lives (t(1/2)) of DBP and DEHP ranged from 1.6 to 2.9 d and 5.0 to 8.3d, respectively. The addition of yeast extract (5mg/L), hydrogen peroxide (1mg/L), brij 35 (91 µM), humic acid (0.5 g/L), cellulose (0.96 mg/L), and sodium chloride (1%) enhanced PAE aerobic degradation. Sediment samples were separated into fractions with various particle size ranges from 0.1-0.45 to 500-2000 µm. Sediment fractions with smaller particle sizes demonstrated higher PAE biodegradation rates. Of the microorganism strains isolated from the mangrove sediment, strains J2, J4, and J8 (all identified as Bacillus sp.) expressed the best biodegrading ability. Our results showed that Bacillus sp. was the dominant bacteria in the process of PAE aerobic degradation in the mangrove sediments.

Anaerobic degradation of nonylphenol in subtropical mangrove sediments.

Nonylphenol (NP) is known as an endocrine disruptor and has consequently drawn much environmental concern. We investigated the effects of various factors on the anaerobic degradation of NP and characterized the structures of microbial communities in mangrove sediments collected at five sites along the Tanshui River in northern Taiwan. NP anaerobic degradation rate constants (k(1)) and half-lives (t(1/2)) ranged from 0.008 to 0.0131/day and 53.3 to 86.6 days, respectively. The addition of NaCl (1%, 2%), zero-valent iron (10 g/L), humic acid (0.5 g/L), cellulose (0.96 mg/L), brij 30 (55 microM) and brij 35 (91 microM) enhanced NP anaerobic degradation. However, the addition of NaCl (3%), acetate (20mM), lactate (20mM), pyruvate (20mM), and humic acid (5 g/L) inhibited NP anaerobic degradation. Sulfate-reducing bacteria, methanogen, and eubacteria are involved in the degradation of NP, sulfate-reducing bacteria being a major component of the sediment. Our results also show that the addition of various substrates changed the microbial community in mangrove sediments. Also noted was the presence of 2-butyl-1-octanol, an intermediate product resulting from the anaerobic degradation of NP accumulated in the sediments.

Anaerobic degradation of phenanthrene and pyrene in mangrove sediment.

This study investigated the anaerobic degradation of the polycyclic aromatic hydrocarbons (PAHs) phenanthrene and pyrene in mangrove sediment from Taiwan. The anaerobic degradation of PAH was enhanced by the addition of acetate, lactate, pyruvate, sodium chloride, cellulose, or zero-valent iron. However, it was inhibited by the addition of humic acid, di-(2-ethylhexyl) phthalate (DEHP), nonylphenol, or heavy metals. Of the microorganism strains isolated from the sediment samples, we found that strain MSA3 (Clostridium pascui), expressed the best ability to biodegrade PAH. The inoculation of sediment with the strain MSA3 could enhance PAH degradation.

Dechlorination of polychlorinated biphenyl congeners by anaerobic microorganisms from river sediment.

The microbial dechlorination of seven kinds of polychlorinated biphenyls (PCBs) by anaerobic microorganisms from river sediment was investigated. Dechlorination rates were found to be affected by the chlorine level of PCB congeners; dechlorination rates decreased as chlorine levels increased. Dechlorination rates were fastest under methanogenic conditions and slowest under nitrate-reducing conditions. The addition of individual electron donors (acetate, pyruvate, and lactate) enhanced the dechlorination of PCB congeners under methanogenic and sulfate-reducing conditions but delayed the dechlorination of PCB congeners under nitrate-reducing conditions. PCB congener dechlorination also was delayed by the addition of various polycyclic aromatic hydrocarbons (PAHs) under three reducing conditions and by surfactants, such as brij30, triton SN70, and triton N101. The results suggest that methanogen, sulfate-reducing bacteria, and nitrate-reducing bacteria all are involved in the dechlorination of PCB congeners.