Biodegradation of nicotine by a novel Strain Shinella sp. HZN1 isolated from activated sludge

The nicotine-degrading bacterium HZN1 was isolated from activated sludge and identified as Shinella sp. based on its physiological characteristics and analysis of 16S rDNA gene. Strain HZN1 is capable of using nicotine as the sole carbon source in the mineral salts medium. The optimum temperature and pH for strain HZN1 growth and nicotine degradation were 30°C and 7.0, respectively. It could degrade approximately 100 % of 0.5 g L(-1) of nicotine within 9 h. Three intermediate metabolites were produced by the strain HZN1 and identified as cotinine, myosmine and nicotyrine using gas chromatography-mass spectrometry. This is the first report of nicotine-degrading strain from the genus of Shinella. The results showed that strain HZN1 could be potentially employed in bioremediation of nicotine. Our findings would provide a new insight into the biodegradation of nicotine.     

Studies on Biodegradation of Nicotine by Arthrobacter sp. Strain HF-2

A nicotine-degrading bacterium, strain HF-2, was isolated from tobacco waste-contaminated soil and identified as a member of Arthrobacter sp. based on morphology, physiological tests, 16S rDNA sequence and phylogenetic characteristics. At thermal denaturation test indicated that the G + C mol% of strain HF-1 was 63.5. The relationship between the growth of the isolate and the nicotine degradation suggested that strain HF-2 could utilize nicotine as sole sources of carbon, nitrogen and energy. Blue pigment was observed during the nicotine degradation by strain HF-2. The isolate grew well at 20 to 33°C, initial pH 6.5 to 8.0 and 0.5 to 2.0 g L^?1 of nicotine concentration in the nicotine inorganic salt media. The maximum growth and nicotine degradation occurred at 30°C, initial pH 7.0 and 0.7 g·L^?1 of nicotine concentration in media under natural incubation condition. Strain HF-2 could degrade 100% of nicotine under the optimized incubation conditions for 43 h. The concentrations of nicotine were monitored by high performance liquid chromatography. This study demonstrates Arthrobacter sp. strain HF-2 had a great ability to degrade nicotine, and it may be available for the application to the bioremediation of environments contaminated by tobacco waste.     

Studies on biodegradation of nicotine by Arthrobacter sp. strain HF-2

A nicotine-degrading bacterium, strain HF-2, was isolated from tobacco waste-contaminated soil and identified as a member of Arthrobacter sp. based on morphology, physiological tests, 16S rDNA sequence and phylogenetic characteristics. At thermal denaturation test indicated that the G + C mol% of strain HF-1 was 63.5. The relationship between the growth of the isolate and the nicotine degradation suggested that strain HF-2 could utilize nicotine as sole sources of carbon, nitrogen and energy. Blue pigment was observed during the nicotine degradation by strain HF-2. The isolate grew well at 20 to 33 degrees C, initial pH 6.5 to 8.0 and 0.5 to 2.0 g L-1 of nicotine concentration in the nicotine inorganic salt media. The maximum growth and nicotine degradation occurred at 30 degrees C, initial pH 7.0 and 0.7 g.L-1 of nicotine concentration in media under natural incubation condition. Strain HF-2 could degrade 100% of nicotine under the optimized incubation conditions for 43 h. The concentrations of nicotine were monitored by high performance liquid chromatography. This study demonstrates Arthrobacter sp. strain HF-2 had a great ability to degrade nicotine, and it may be available for the application to the bioremediation of environments contaminated by tobacco waste.     

菌源对多环芳烃降解菌的筛选及降解性能的影响

高效降解菌的筛选对利用生物修复技术有效去除环境中的多环芳烃具有重要意义。分别以石油污染土壤和焦化废水活性污泥为菌源,分离出芘降解菌和混合PAHs(菲、荧蒽和芘)降解菌共14株并对其降解性能进行对比研究。结果表明,筛选得到的菌株分别属于9个菌属,其中2种菌源共有的菌属为Mycobacterium sp.、Ralstonia sp.和Shinella sp.。芘和PAHs的高效降解菌(CP16和CM32)均属于分支杆菌属(Mycobacterium),来源于焦化废水活性污泥;菌株CP16对芘(50mg/L)的7 d降解率为74.99%,CM32对PAHs(菲50 mg/L、荧蒽和芘各10 mg/L)的7 d降解率为100%。因此,以焦化废水活性污泥为菌源更有利于获得高效的多环芳烃降解菌。     

Degradation of chlorpyrifos by an endophytic bacterium of the Sphingomonas genus (strain HJY) isolated from Chinese chives (Allium tuberosum)

The degradation of chlorpyrifos (CP) by an endophytic bacterial strain (HJY) isolated from Chinese chives (Allium tuberosum Rottl. ex Spreng) was investigated. Strain HJY was identified as Sphingomonas sp. based on morphological, physiological, and biochemical tests and a 16S rDNA sequence analysis. Approximately 96% of 20 mg L^?1 CP was degraded by strain HJY over 15 days in liquid minimal salts medium (MSM). The CP degradation rate could also be increased by glucose supplementation. The optimal conditions for the removal of 20 mg L^?1 CP by strain HJY in MSM were 2% inoculum density, pH 6.0, and 30–35°C. The CP degradation rate constant and half-life were 0.2136 ± 0.0063 d^?1 and 3.2451 ± 0.0975 d, respectively, under these conditions, but were raised to 0.7961 ± 0.1925 d^?1 and 0.8707 ± 0.3079 d with 1% glucose supplementation. The detection of metabolic products and screening for degrading genes indicated that O,O-diethyl O-3,5,6-trichloropyridinol was the major degradation product from CP, while it was likely that some functional genes were undetected and the mechanism responsible for CP degradation by strain HJY remained unknown. Strain HJY is potentially useful for the reduction of CP residues in Chinese chives and may be used for the in situ phytoremediation of CP.     

Biodegradation of acetochlor by a newly isolated Achromobacter sp. strain D-12

A highly effective acetochlor-degrading bacterial strain (D-12) was isolated from the soil of a pesticide factory. The strain was identified as Achromobacter sp. based on its 16S rRNA gene sequence. The strain D-12 optimally degrades acetochlor at a pH of 7.0 and a temperature of 30°C in a mineral salts medium (MSM). Approximately 95% of acetochlor was degraded by the stain treated at a concentration of 10 mg L^?1 after 5 days of incubation. A chiral high performance liquid chromatography (HPLC) system was used to study the enantioselectivity during the process. However, no obvious enantioselective biodegradation was observed. The primary biodegradation acetochlor products were identified by high-performance liquid chromatography-mass spectroscopy (HPLC-MS) and gas chromatography-mass spectrometry (GC-MS). The results indicated that the strain D-12 could be applied in the bioremediation of an acetochlor-polluted environment.     

Biodegradation of imidacloprid in liquid media by an isolated wastewater fungus Aspergillus terreus YESM3

In the present study, a new fungal strain capable of imidacloprid degradation was isolated from agricultural wastewater drain. The fungal strain of YESM3 was identified as Aspergillus terreus based on ITS1-5.8S rDNA-ITS2 gene sequence by PCR amplification of a 500 bp sequence. Screening of A. terreus YESM3 to the insecticide imidacloprid tolerance was achieved by growing fungus in Czapek Dox agar for 6 days at 28°C. High values (1.13 and 0.94 cm cm^?1) of tolerance index (TI) were recorded at 25 and 50 mg L^?1 of imidacloprid, respectively in the presence and absence of sucrose. However, at 400 mg L^?1 the fungus did not grow. Effects of the imidacloprid concentration, pH, and inoculum size on the biodegradation percentage were tested using Box–Behnken statistical design and the biodegradation was monitored by HPLC analysis at different time intervals. Box–Behnken results indicated that optimal conditions for biodegradation were at pH 4 and two fungal discs (10 mm diameter) in the presence of 61.2 mg L^?1 of imidacloprid. A. terreus YESM3 strain was capable of degrading 85% of imidacloprid 25 mg L^?1 in Czapek Dox broth medium at pH 4 and 28°C for 6 days under static conditions. In addition, after 20 days of inoculation, biodegradation recorded 96.23% of 25 mg L^?1 imidacloprid. Degradation kinetics showed that the imidacloprid followed the first order kinetics with half-life (t₅₀) of 1.532 day. Intermediate product identified as 6-chloronicotinic acid (6CNA) as one of the major metabolites during degradation of imidacloprid by using HPLC. Thus, A. terreus YESM3 showed a potential to reduce pollution by pesticides and toxicity in the effected environment. However, further studies should be conducted to understand the biodegradation mechanism of this pesticide in liquid media.     

一株苯酚降解菌的筛选及降解动力学特性

利用富集驯化的培养方法,从首钢焦化厂废水处理系统中的二沉池出水中,分离筛选出一株能够高效降解苯酚的菌株B3对其16S rDNA序列进行分析,并选择Monod方程和Andrews方程分别研究该菌在不同苯酚浓度条件下的降酚动力学模式。结果表明,B3为蜡状芽孢杆菌(Bacillus cereus);苯酚浓度较低时,苯酚对菌株的生长基本不产生抑制作用,用Monod模型对B3降酚动力学过程进行拟合,其动力学参数V max=0.03 h-1,K s=25.53 mg/L;苯酚浓度较高时,按照Andrews模型对B3降酚动力学过程进行非线性最小二乘曲线拟合,其动力学参数V max=0.08 h-1,K s=147.52 mg/L,K i=384.96 mg/L。根据动力学方程,推论菌株B3降解对于浓度238.30 mg/L的苯酚具有最佳降解效果。     

Effects of Bacillus subtilis on the growth,colony maintenance,and attached bacterial community composition of colonial cyanobacteria

In freshwater aquaculture ponds, application of algicidal Bacillus is a promising way in the control of cyanobacterial blooms. To best understand Bacillus algicidal characters and mechanisms in the field, different-sized colonial cyanobacteria were isolated from an aquaculture pond, and the effects of B. subtilis on their growth, colony maintenance, and colony-attached bacterial community composition were investigated. The results showed that B. subtilis could inhibit the growth of colonial cyanobacteria. Bigger-sized colonies isolated from the field could spontaneously disintegrate into smaller-sized colonies in the laboratory. Algicidal B. subtilis could accelerate the disintegration of colonies and decrease colony size. B. subtilis not only decreased the colony-attached bacterial community diversity but also changed its composition. B. subtilis increased the relative abundances of some attached bacterial genera, including Pseudomonas, Shewanella, Bacillus, Shinella, Rhizobium, and Ensifer. These bacteria with algicidal, microcystin-degrading, and flocculating activities might be an important contributor to algicidal effects of B. subtilis on colonial cyanobacteria.

     

Evaluation of Saccharomyces cerevisiae strains as probiotic agent with aflatoxin B1 adsorption ability for use in poultry feedstuffs

In this study the aflatoxin B₁ (AFB₁) removal capacity, the tolerance to salivary and gastrointestinal conditions, autoaggregation and coaggregation with pathogenic bacteria of Saccharomyces cerevisiae strains isolated from broiler feces, were evaluated. Only four of twelve isolated strains were identified as Saccharomyces cerevisiae using molecular techniques. The results obtained in AFB₁ binding studies indicated that the amount of AFB₁ removed was both strain and mycotoxin-concentration dependent. Therefore, a theoretical model was applied in order to select the most efficient strain to remove AFB₁ in a wide range of mycotoxin concentration. The results indicated that S. cerevisiae 08 and S. cerevisiae 01 strains were the most efficient microorganisms in the mycotoxin removal. Viability on simulated salivary and gastrointestinal conditions was investigated and S. cerevisiae 08 strain showed the best results, achieving 98% of total survival whereas S. cerevisiae 01 reached only 75%. Autoaggregation and coaggregation assays showed S. cerevisiae 08 as the most appropriate strain, mainly because it was the unique strain able to coaggregate with the four bacterial pathogens assayed. Consequently, S. cerevisiae 08 is the best candidate for future in vivo studies useful to prevent aflatoxicosis. Further quantitative in vitro and in vivo studies are required to evaluate the real impact of yeast-binding activity on the bioavailability of AFB₁ in poultry. However, this study could be useful in selecting efficient strains in terms of AFB₁ binding and provide an important contribution to research into microorganisms with potential probiotic effects on the host.     

Benzoate degradation by Rhodococcus opacus 1CP after dormancy: Characterization of dioxygenases involved in the process

The process of benzoate degradation by strain Rhodococcus opacus 1CP after a five-year dormancy was investigated and its peculiarities were revealed. The strain was shown to be capable of growth on benzoate at a concentration of up to 10 g L^?1. The substrate specificity of benzoate dioxygenase (BDO) during the culture growth on a medium with a low (200–250 mg L^?1) and high (4 g L^?1) concentration of benzoate was assessed. BDO of R. opacus 1CP was shown to be an extremely narrow specificity enzyme. Out of 31 substituted benzoates, only with one, 3-chlorobenzoate, its activity was higher than 9% of that of benzoate. Two dioxygenases, catechol 1,2-dioxygenase (Cat 1,2-DO) and protocatechuate 3,4-dioxygenase (PCA 3,4-DO), were identified in a cell-free extract, purified and characterized. The substrate specificity of Cat 1,2-DO isolated from cells of strain 1CP after the dormancy was found to differ significantly from that of Cat 1,2-DO isolated earlier from cells of this strain grown on benzoate. By its substrate specificity, the described Cat 1,2-DO was close to the Cat 1,2-DO from strain 1CP grown on 4-methylbenzoate. Neither activity nor inhibition by protocatechuate was observed during the reaction of Cat 1,2-DO with catechol, and catechol had no inhibitory effect on the reaction of PCA 3,4-DO with protocatechuate.     

Effect of Pseudomonas sp. HF-1 inoculum on construction of a bioaugmented system for tobacco wastewater treatment: analysis from quorum sensing

To better construct a bioaugmented system for tobacco wastewater treatment, activated sludge was inoculated with different concentrations of the nicotine-degrading bacterium Pseudomonas sp. HF-1. The results showed that inoculum concentrations of 0.55?±?0.01 and 1.10?±?0.03 mg/g (dry weight of strain HF-1/dry weight of activated sludge) were best to ensure strain HF-1 survival and successful bioaugmentation. The release pattern of autoinducer (AI) for quorum sensing in the bioaugmented system was also investigated. During the period of HF-1 inoculation, compared with failed bioaugmented systems, AI-2 was significantly increased in the successful systems, suggesting that AI-2-mediated bacterial communication played an important role in the colonization of HF-1. When inoculation of strain HF-1 was stopped, the amount of AI-2 decreased and leveled out in all systems. Notably, there was a greater than threefold increase of short-chain AHLs in failed bioaugmented systems, but no increase in successful ones, implying that the fluctuation of short-chain AHLs could be an indicator of the failure of bioaugmentation. Thus, AI-2-mediated quorum sensing could be implemented to facilitate HF-1 colonization.     

Mineralization of p-nitrophenol by a new isolate Arthrobacter sp. Y1

Arthrobacter sp. Y1, capable of metabolizing p-nitrophenol (PNP) as the sole carbon, nitrogen and energy source was isolated from activated sludge. The bacterium could tolerate concentrations of PNP up to 600 mg L^{? 1}, and degradation of PNP was achieved within 120 h of incubation. PNP and its metabolites were analyzed by high performance liquid chromatography (HPLC). The metabolite formed indicated that the organism followed the 4-nitrocathechol (4-NC) pathway for metabolism of this compound. The relevant degrading-enzyme was extracellular. Addition of other carbon source (glucose 0～ 30 g L^{? 1}) led to accelerated degradation. If the glucose concentration exceeded 30 g L^{? 1}, however, degradation was repressed. Spectrophotometry assay of the nitrite and genotoxic study showed that strain Y1 could detoxify PNP. Therefore, the present study may provide a basis for the development of the bioremediation strategies to remedy the pollutants in the environment.     

Composition and emissions of VOCs in main- and side-stream smoke of research cigarettes

It is well known that mainstream (MS) and sidestream (SS) cigarette smoke contains a vast number of chemical substances. Previous studies have emphasized SS smoke rather than MS smoke to which smokers are exposed, and most have used chamber tests that have several disadvantages such as wall losses. Emissions from standard research cigarettes have been measured, but relatively few constituents have been reported, and only the 1R4F (low nicotine) cigarette type has been tested. This study provides a comprehensive characterization of total, MS and SS smoke emissions for the 1R5F (ultra low nicotine), 2R4F (low nicotine), and 1R3F (standard nicotine) research cigarettes research cigarettes, including emission factors for a number of toxic compounds (e.g., benzene) and tobacco smoke tracers (e.g., 2,5-dimethyl furan). Emissions of volatile organic compounds (VOCs) and particulate matter (PM) are quantified using a dynamic dilution emission measurement system that is shown to produce accurate, rapid and reproducible results for over 30 VOCs and PM. SS and MS emissions were accurately apportioned based on a mass balance of total emissions. As expected, SS emissions greatly exceeded MS emissions. The ultra low nicotine cigarette had lower emissions of most VOCs compared to low and standard nicotine cigarettes, which had similar emissions. Across the three types of cigarettes, emissions of benzene (296–535 μg cig⁻¹), toluene (541–1003 μg cig⁻¹), styrene (90–162 μg cig⁻¹), 2-dimethyl furan (71–244 μg cig⁻¹), naphthalene (15–18 μg cig⁻¹) and other VOCs were generally comparable to or somewhat higher than literature estimates using chamber tests.     

Effects of the herbicide molinate on the metabolic activities of a degradative Streptomyces griseus strain

Abstract

Cometabolic degradation of the herbicide molinate was tested using two microorganisms, Arthrobacter sp., strain M3 and Streptomyces griseus strain M2; the latter classified on the basis of the presence of the enzymatic cofactor SF‐420. The strains M3 and M2, inoculated in a basic salts medium with glucose as carbon source and added with 100 mg L^‐1 of molinate, degraded respectively 35 and 51% of the herbicide in 36 days.

Increasing concentrations of molinate, ranging from 50 to 200 mg L^‐1 in glucose medium, did not affect the final ATP yield of the strain M2, but decreased the final growth yield and the ATP synthesis rate. Moreover, the onset of coenzyme SF‐420 synthesis was progressively delayed.

In contrast, surprisingly, SF‐420 final yield and production rate were increased by progressive increasing concentrations of molinate in the mineral medium.     

Biodegradation of 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) by using Serratia marcescens NCIM 2919

A solvent tolerant bacterium Serratia marcescens NCIM 2919 has been evaluated for degradation of DDT (1,1,1-trichloro-2,2-bis (4-chlorophenyl) ethane). The bacterium was able to degrade up to 42% of initial 50 mg L^?1 of DDT within 10 days of incubation. The highlight of the work was the elucidation of DDT degradation pathway in S. marcescens. A total of four intermediates metabolites viz. 2,2-bis (chlorophenyl)-1,1-dichloroethane (DDD), 2,2-bis (chlorophenyl)-1,1-dichloroethylene (DDE), 2,2-bis (chlorophenyl)-1-chloroethylene (DDMU), and 4-chlorobenzoic acid (4-CBA) were identified by GC-Mass and FTIR. 4-CBA was found to be the stable product of DDT degradation. Metabolites preceding 4-CBA were not toxic to strain as reveled through luxuriant growth in presence of varying concentrations of exogenous DDD and DDE. However, 4-CBA was observed to inhibit the growth of bacterium. The DDT degrading efficiency of S. marcescens NCIM 2919 hence could be used in combination with 4-CBA utilizing strains either as binary culture or consortia for mineralization of DDT. Application of S. marcescens NCIM 2919 to DDT contaminated soil, showed 74.7% reduction of initial 12.0 mg kg^?1 of DDT after 18-days of treatment.     

不适宜生长条件对混菌降解苯胺的影响

分别从台州和衢州某化工厂的好氧池中分离筛选得到2株苯胺降解菌TZ1和JH1,经16S rDNA测序鉴定为Comamonas sp.TZ1和Pseudomonas sp.JH1,均具有较强的苯胺降解能力,培养24 h后,可使初始浓度为800 mg/L的苯胺去除率达到96.4%~98.4%。在此基础上,按体积比1∶1将2株菌液进行混合构建了混合菌体系,进而对比考察了苯胺初始浓度、pH、盐度和重金属等环境因子对单一菌和混合菌生长量及降解苯胺效果的影响,重点探讨混合菌对不适宜生长环境的适应性及其对苯胺的降解特性。通过单一菌和混合菌对比实验发现,在适宜苯胺初始浓度、pH和盐度条件下,混合菌的生长量略高于单一菌;在不适宜生长的高浓度苯胺、pH和盐度条件下,混合菌也表现出了更强的适应性和苯胺矿化能力。Zn2+和Cr6+耐受实验则表明,对于Cr6+,混合菌表现出了更强的耐受能力,而对于Zn2+并没有表现出更强的耐受能力。     

Characterization of isolated yeast growth response to methionine analogs

Methionine is one of the first limiting amino acids in poultry nutrition. The use of methionine-rich natural feed ingredients, such as soybean meal or rapeseed meal may lead to negative environmental consequences. Amino acid supplementation leads to reduced use of protein-rich ingredients. The objectives of this study were isolation of potentially high content methionine-containing yeasts, quantification of methionine content in yeasts and their respective growth response to methionine analogs. Minimal medium was used as the selection medium and the isolation medium of methionine-producing yeasts from yeast collection and environmental samples, respectively. Two yeasts previously collected along with six additional strains isolated from Caucasian kefir grains, air-trapped, cantaloupe, and three soil samples could grow on minimal medium. Only two of the newly isolated strains, K1 and C1, grew in minimal medium supplied with either methionine analogs ethionine or norleucine at 0.5% (w/v). Based on large subunit rRNA sequences, these isolated strains were identified as Pichia udriavzevii/Issatchenkia orientalis. P. kudriavzevii/I. orentalis is a generally recognized as a safe organism. In addition, methionine produced by K1 and C1 yeast hydrolysate yielded 1.3 ± 0.01 and 1.1 ± 0.01 mg g^?1 dry cell. Yeast strain K1 may be suitable as a potential source of methionine for dietary supplements in organic poultry feed but may require growth conditions to further increase their methionine content.     

The toxicity of copper, cadmium and zinc to four different Hydra (Cnidaria: Hydrozoa)

An acute toxicity study of three metals to Hydra species carried out using two different assessment methods, (i) determination of the LC50 and (ii) measurement of progressive morphological changes, demonstrated that relative toxicity decreased from copper to cadmium with zinc the least toxic for all species. The latter method revealed more details of the effect on Hydra in terms of physical damage to the polyp but both methods indicated that H. viridissima was more sensitive to copper and cadmium than H. vulgaris¹ (Zurich strain, male clone), H. vulgaris² (a dioecious strain reproducing sexually and asexually) and H. oligactis (dioecious, reproducing sexually and asexually). The responses to zinc were similar for all Hydra. The possible role of metabolic interactions between H. viridissima and its symbiotic green algae in contributing to the greater sensitivity of this polyp is discussed.     

Evidence for enhanced dissipation of chlorpyrifos in an agricultural soil inoculated with Serratia rubidaea strain ABS 10

The insecticide ¹⁴C-chlorpyrifos was found mineralized in a Tunisian soil with repeated exposure to it. From this soil, a bacterial strain was isolated that was able to grow in a minimal salt medium (MSM) supplemented with 25 mg L^?1 of chlorpyrifos. It was characterized as Serratia rubidaea strain ABS 10 using morphological and biochemical analyses, as well as 16S rRNA sequencing. In a liquid culture, the S. rubidaea strain ABS 10 was able to dissipate chlorpyrifos almost entirely within 48 h of incubation. Although the S. rubidaea strain ABS 10 was able to grow in an MSM supplemented with chlorpyrifos and dissipate it in a liquid culture, it was not able to mineralize ¹⁴C-chlorpyrifos. Therefore, it can be concluded that the dissipation capability of this bacteria might be attributed to its capacity to adsorb CHL. It can also be ascribed to other reasons such as the formation of biogenic non-extractable residues. In both non-sterile and sterile soil inoculated with S. rubidaea strain ABS 10, chlorpyrifos was more rapidly dissipated than in controls with DT₅₀ of 1.38 and 1.05 days, respectively.