The Preliminary Study on Screening and Application of Phthalic Acid-Degrading Bacteria

Phthalic acid is a main pollutant, which is also an important reason for the continuous cropping effect of tobacco. In order to degrade the phthalic acid accumulated in the environment and relieve the obstacle effect of tobacco continuous cropping caused by the accumulation of phthalic acid in the soil. In this study, phthalate degrading bacteria B3 is screened from continuous cropping tobacco soil. The results of biochemical identification and 16sDNA comparison show that the homology between degrading bacterium B3 and Enterobacter sp. is 99%. At the same time, the growth of Enterobacter hormaechei subsp. B3 and the degradation of phthalic acid under different environmental conditions are studied. The results show that the environment with a temperature of 30˚C, PH of 7, and inoculation amount of not less than 1.2%, which is the optimal growth conditions for Enterobacter sp. B3. In an environment with a concentration of phthalic acid not exceeding 500 mg/L, Enterobacter sp. B3 has a better effect on phthalic acid degradation, and the degradation rate can reach 77% in 7 d. The results of indoor potting experiments on tobacco show that the degradation rate of phthalic acid by Enterobacter B3 in the soil is about 45%, which can reduce the inhibitory effect of phthalic acid on the growth of tobacco seedlings. This study enriches the microbial resources for degrading phthalic acid and provides a theoretical basis for alleviating tobacco continuous cropping obstacles.

Cumulative Toxicity of Residue Degradation Products of Penicillin Bacteria

[Objective] The research aimed to discuss the accumulation of toxic slag of penicillin bacteria residue degradation products and explore its ability to meet the aquaculture industry as a protein feed into development and utilization conditions.[Method] Through the sub-acute toxicity tests in mice strains,which were fed by different doses of penicillin bacteria residue degradation products （3% and 6%） under continuous observation of 15 weeks,recording a weekly mouse weight and death,and sampling executed after the test,animal liver and kidney function were blood test,taking heart,liver,spleen,kidney weighing,as well as liver and kidney pathology observed in the optical microscope.[Result] There were no significant differences （P 0.05） between the test group mice body weight,mortality and liver and kidney function and the control group within 15 weeks.Low-dose test group could be seen the liver cells,renal tubular epithelial nuclei broken,and a small number of liver and kidney cells with mild edema.High-dose test group could be seen in liver tissue of mice nuclei fragmentation and a fat droplets,the majority of liver cells,edema,and only a small number of liver cells,there were no significant changes.Renal portal area showed inflammatory cell infiltration,renal tubular epithelial cells,edema and necrosis.[Conclusion] In this experimental condition,the degradation products of penicillin bacteria residue played a mild toxcity on organ parenchymal cells in mice.

Cultivating High Efficient Bacteria of Degrading Pulping Wastewater by Ultraviolet Mutagenic Technology

Instead of pure bacteria, induction mutation of activated sludge by ultraviolet (Uv) was studied and used to treat pulping wastewater by continuous- flow. The result showed the mutagenic activated sludge had remarkable effect and application potential in pulping wastewater treatment. Comparing with common activated sludge, the mutagenic activated sludge was more suitable for lignose decomposition and had high decomposing efficiency.

Application of Uniform Design and SVR for Culture Medium Optimization of Organophosphorus Degradation Bacteria

In order to improve the degradation activity of organophosphorus degradation bacterial strains, uniform design and support vector regression(SVR) are used to optimize the formula of mineral salt medium for the strains based on single factor experiment. The formula being used after SVR optimization is glucose 1.24 g/L,NH4 NO3 0.73 g, KH2 PO4 0.96 g, KCl 0.50 g, MgSO4·7 H2 O 0.50 g, MnSO4 0.02 g, CaCl2 0.04 g, distilled water 1 000 mL;the highest degradation rate is obtained at pH7.0(SVR, 76.68%), superior to that of the QSAR model(74.67%). The formula optimization method based on uniform design and SVR is very promising to apply in various multiple-factor and multiple-level formula optimization experiments.

Biodegradation Characteristics of Environmental Endocrine Disruptor Di-n-butyl Phthalate

Objective The biodegradation characteristics of di-n-butyl phthalate (DBP), an environmental endocrine disruptor, were studied by the method of dominant bacteria and immobilized microorganisms. Methods Taking DBP as the only carbon source to acclimatize the collected activated sludge, the concentration of DBP increased progressively in the process of acclimatization. Plate streaking was used to separate 1 strain of the degradation dominant bacteria after acclimatization. Better conditions to degrade DBP by the bacterium could be obtained through orthogonal experiments and the bacterium was identified. Then the acclimated activated sludge was made to immobilize the microorganism using polyvinyl alcohol as entrapment agent. The immobilized microorganism degraded DBP at different conditions. Results The appropriate conditions to degrade DBP by the dominant bacteria were: degradation time, 32 h; DBP concentration, 200 mg/L; rate of shaking incubator, 100 r/min; pH, 7 and temperature, 30℃. DBP could be degraded by more than 95% under such conditions. The bacteria were identified as pseudomonas. The proliferated immobilized microorganisms degraded DBP more effectively and more adapted to temperature and pH than the free acclimated activated sludge. Conclusion One strain of DBP degradation dominant bacteria was separated from the acclimatized activated sludge. It could grow with DBP as the only carbon source and energy, and degraded DBP effectively. After having been immobilized and proliferated, the dominant bacteria could keep a higher biological activity and degrade DBP more effectively than activated sludge.

Isolation and characterization of diesel degrading bacteria, Sphingomonas sp. and Acinetobacter junii from petroleum contaminated soil

Two indigenous bacteria of petroleum con- taminated soil were characterized to utilize diesel fuel as the sole carbon and energy sources in this work. 16S rRNA gene sequence analysis identified these bacteria as Sphingomonas sp. and Acinetobacterjunii. The ability to degrade diesel fuel has been demonstrated for the first time by these isolates. The results of IR analyses showed that Sphingomonas sp. VA1 and A. junii VA2 degraded up to 82.6% and 75.8% of applied diesel over 15days, respectively. In addition, Sphingomonas sp. VA1 pos- sessed the higher cellular hydrophobicities of 94% for diesel compared to 81% by A. junii VA2. The isolates Sphingomonas sp. VA1 and A. junii VA2 exhibited 24% and 18%, respectively emulsification activity. This study reports two new diesel degrading bacterial species, which can be effectively used for bioremediation of petroleum contaminated sites.

Carrageenan oligosaccharides and associated carrageenan-degrading bacteria induce intestinal inflammation in germ-free mice

Carrageenans(CGNs)are widely used in foods and pharmaceuticals although their safety remains controversial.To investigate the effects of CGNs and CGN-degrading bacteria in the human colon,we screened for CGN degradation by human fecal microbiota,and for inflammatory response to CGNs and/or CGN-degrading bacteria in germ free mice.Thin-layer chromatography indicated that high molecular weight(MW)CGNs(!100 kDa)remained undegraded in the presence of human fecal microbiota,whereas low MW CGNs,i.e.,k-carrageenan oligosaccharides(KCO,~4.5 kDa)were degraded when exposed to seven of eight human fecal samples,although sulfate groups were not removed during degradation.Bacteroides xylanisolvens and Escherichia coli isolates from fecal samples apparently degraded KCO synergistically,with B.xylanisolvens serving as the primary degrader.Combined treatment of KCO with KCO-degrading bacteria led to greater pro-inflammatory effects in the colon and rectum of germ-free mice than either KCO or bacteria alone.Similarly,p-p38-,CD3-,and CD79a-positive immune cells were more abundant in combined treatment group mice than in either single treatment group.Our study shows that KCO-degrading bacteria and the low MW products of KCO can promote proinflammatory effects in mice,and represent two key markers for evaluating CGN safety in foods or medicines.

Isolation and characterization of endosulfan-degrading bacteria from contaminated agriculture soils

Objective:To isolate and characterize endosulfan-degrading bacteria from Kerman pistachio orchards.Methods:Endosulfan-degrading bacteria were enriched in Bushnell-Hass medium.Identification and sequencing of prevalent degrading strains was performed by using PCR based on amplifying 16S rDNA.Results:The results showed that the soils of pistachio orchards have some degrading bacteria that are suitable for elimination of endosulfan from soils and the environment.Four endosulfandegrading bacteria strains belong to Achromobacter xylosoxidans(strain EN3),Pseudomonas azotoformans(strain EN4),Pseudomonas brassicacearum(strain EN7)and Pseudomonas thivervalensis(strain EN8),respectively.The best degrading strain(EN7),up to 100 mg/L,illustrated a good growth,whereas the growth was reduced in concentration higher than 100 mg/L.The results of gas chromatography confirmed the decomposition of organic pesticide by degrading-bacteria.Conclusions:By using these strains and other biological reclamation methods we can eliminate bio-environmental problems.

Biodegradation of Phthalate Esters by a Newly Isolated Acinetobacter sp. Strain LMB-5 and Characteristics of Its Esterase

Phthalate esters （PAEs） are extensively applied in industry, and they migrate to environment during the process of production, employ, and treatment and axe difficult to be degraded in nature. However, some microorganisms could use them as the carbon source to growth. In this study, an Acinetobacter sp. strain LMB-5, capable of utilizing PAEs, was isolated from a vegetable greenhouse soil. The degradation capability of strain LMB-5 was also investigated by incubation in mineral salt medium containing different PAEs, dimethyl phthalate （DMP）, diethyl phthalate （DEP）, di-n-butyl phthalate （DBP）, and di-（2-ethylhexyl） phthalate （DEHP）. The strain could grow well with DMP, DEP, DBP, and DEHP. When the concentration of DBP increased from 100 to 400 mg L-1, the half-life extended from 9.5 to 15.5 h. In the concentration range of DBP, the degradation ability of strain LMB-5 could be described by first-order kinetics. During the biodegradation of DBP, three intermediates, 1,2-benzenedicaxboxylic acid,butyl methyl ester, DMP, and phthalic acid （PA） were detected, and the proposed pathway of DBP was identified. By analysis of bioinformatics, one esterase was cloned from the genome of LMB-5 and expressed in Escherichia coll. It displayed an ability to break the ester bonds of DBP. The enzyme exhibited maximal activity at pH 7.0 and 40 ℃ with DBP as the substrate. It was activated by Cu2＋ and Fe3＋ and had a high activity in the presence of low concentrations of methanol or dimethylsulfoxide （each 10%, volume：volume）. The Acinetobacter sp. strain LMB-5 may make a contribution to the remediation of soils polluted by PAEs in the future.

Molecular characterization of methanogenic microbial communities for degrading various types of polycyclic aromatic hydrocarbon

Knowledge on methanogenic microbial communities associated with the degradation of polycyclic aromatic hydrocarbons(PAHs)is crucial to developing strategies for PAHs bioremediation.In this study,the linkage between the type of PAHs and microbial community structure was fully investigated through 16 S rRNA gene sequencing on four PAH-degrading cultures.Putative degradation products were also detected.Our results indicated that naphthalene(Nap)/2-methylnaphthalene(2-Nap),phenanthrene(Phe)and anthracene(Ant)sculpted different microbial communities.Among them,Nap and 2-Nap selected for similar degrading bacteria(i.e.,Alicycliphilus and Thauera)and methanogens(Methanomethylovorans and Methanobacterium).Nap and 2-Nap were probably activated via carboxylation,producing 2-naphthoic acid.In contrast,Phe and Ant shaped different bacterial and archaeal communities,with Arcobacter and Acinetobacter being Phe-degraders and Thiobacillus Ant-degrader.Methanogenic archaea Methanobacterium and Methanomethylovorans predominated Phe-degrading and Ant-degrading culture,respectively.These findings can improve our understanding of natural PAHs attenuation and provide some guidance for PAHs bioremediation in methanogenic environment.

Influence of kaolinite and montmorillonite on benzo[a]pyrene biodegradation by Paracoccus aminovorans HPD-2 and the underlying interface interaction mechanisms

Clay minerals play an important role in biogeochemical cycling.Here,kaolinite and montmorillonite,the two most abundant and widespread clay minerals with typical layered structures,were selected to investigate and compare their effects on the biodegradation of benzo[a]pyrene(BaP)by Paracoccus aminovorans HPD-2 and to investigate the underlying interface mechanisms.Overall,the BaP degradation efficiency was significantly higher 7 d after montmorillonite addition,reaching 68.9%(P<0.05),when compared with that of the control without addition of clay minerals(CK,61.4%);however,the addition of kaolinite significantly reduced the BaP degradation efficiency to 45.8%.This suggests that kaolinite inhibits BaP degradation by inhibiting the growth of strain HPD-2,or its strong hydrophobicity and readily agglomerates in the degradation system,resulting in a decrease in the bio-accessibility of BaP to strain HPD-2.Montmorillonite may buffer some unfavorable factors,and cells may be fixed on the surface of montmorillonite colloidal particles across energy barriers.Furthermore,the adsorption of BaP on montmorillonite may be weakened after swelling,reducing the effect on the bio-accessibility of BaP,thus promoting the biodegradation of BaP by strain HPD-2.The experimental results indicate that differential bacterial growth,BaP bio-accessibility,interface interaction,and the buffering effect may explain the differential effects of the different minerals on polycyclic aromatic hydrocarbon biodegradation.These observations improve our understanding of the mechanisms by which clay minerals,organic pollutants,and degrading bacteria interact during the biodegradation process and provide a theoretical basis for increasing the biodegradation of soil pollutants by native microorganisms under field conditions.