Qingxiang Yang

Evolution of the microbial community in a full-scale printing and dyeing wastewater treatment system

In this study, the dynamics of bacterial, fungal and archaeal populations in two-stage biological processes of a full-scale printing and dyeing wastewater treatment system were traced using cultivation and molecular biological techniques. The enumeration results indicated that bacteria were the dominant population in the system, in which the ratio of fungi to bacteria decreased in all the treatment units, while the ratio of archaea to bacteria increased significantly, especially in samples from the second-stage biological treatment process. PCR-denaturing gradient gel electrophoresis (DGGE) analysis showed that the microbial diversity increased with system running and 64.6% of bacterial, 57.6% of fungal and 38.2% of archaeal populations remained in the system from the seed sludge during system start-up. In spite of variation in the microbial community and composition of the influents, some bacterial species such as Thauera sp. and Xanthomonadaceae were present simultaneously in all the collected samples.

Influence of oxytetracycline on the structure and activity of microbial community in wheat rhizosphere soil

The microbial community composition in wheat rhizosphere was analyzed by detecting colony forming units (CFUs) in agar plates. The total CFUs in rhizosphere were 1.04 x 10(9)/g soil with 9.0 x 10(8)/g bacteria, 1.37 x 10(8)/g actinomyces and 3.6 x 10(6)/g fungi. The 10 dominant bacteria were isolated from wheat rhizosphere and were grouped into genus Bacillus according to their full length 16S rRNA gene sequences. Although belonging to the same genus, the isolated strains exhibited different sensitivities to oxytetracycline. When a series of the rhizosphere soil was exposed under various concentrations of oxytetracycline, the microbial community structure was highly affected with significant decline of CFUs of bacteria and actinomyces (22.2% and 31.7% at 10 mg/kg antibiotic, respectively). This inhibition was clearly enhanced with the increase exposure dosage of antibiotic and could not be eliminated during 30 d incubation. There was no obvious influence of this treatment on fungi population. Among the four soil enzymes (alkaline phosphatase, acidic phosphatase, dehydrogenase and urease), only alkaline phosphatase was sensitive to oxytetracycline exposure with 41.3% decline of the enzyme activity at 10 mg/kg antibiotic and further decrease of 64.3%-80.8% when the dosage over 30 mg/kg.

Effects of glucose on the decolorization of Reactive Black 5 by yeast isolates

The cometabolic roles of glucose were investigated in decolorization of an azo dye, Reactive Black 5, by yeast isolates, Debaryomyces polymorphus and Candida tropicalis. The results indicated that the dye degradation by the two yeasts was highly associated with the yeast growth process and glucose presence in the medium. Color removal of 200 mg dye/L was increased from 76.4% to 92.7% within 60 h to 100% within 18-24 h with the increase of glucose from 5 to 10 g/L, although the activity of manganese dependent peroxidase (MnP) decreased by 2-8 times in this case. Hydrogen peroxide of 233.3 microg/L was detected in 6 h in D. polymorphus culture. The cometabolic functions of glucose and hydrogen peroxide could be also confirmed by the further color removals of 95.8% or 78.9% in the second cycle of decolorization tests in which 7 g glucose/L or 250 microg H2O2/L was superadded respectively together with 200 mg dye/L.

Extracellular enzyme production and phylogenetic distribution of yeasts in wastewater treatment systems

The abilities of yeasts to produce different extracellular enzymes and their distribution characteristics were studied in municipal, inosine fermentation, papermaking, antibiotic fermentation, and printing and dyeing wastewater treatment systems. The results indicated that of the 257 yeasts, 16, 14, 55, and 11 produced lipase, protease, manganese dependant peroxidase (MnP), and lignin peroxidase (LiP), respectively. They were distributed in 12 identified and four unidentified genera, in which Candida rugosa (AA-M17) and an unidentified Saccharomycetales (AA-Y5), Pseudozyma sp. (PH-M15), Candida sp. (MO-Y11), and Trichosporon montevideense (MO-M16) were shown to have the highest activity of lipase, protease, Mnp, and LiP, respectively. No yeast had amylase, cellulose, phytase, or laccase activity. Although only 60 isolates produced ligninolytic enzymes, 249 of the 257 yeasts could decolorize different dyes through the mechanism of biodegradation (222 isolates) or bio-sorption. The types of extracellular enzymes that the yeasts produced were significantly shaped by the types of wastewater treated.

Wastewater treatment by alkali bacteria and dynamics of microbial communities in two bioreactors

In this study, an alkali bacterial consortium was obtained by enrichment cultivation and was used to treat printing and dyeing wastewater (PDW, pH 11-12). The treatment effects and dynamic changes were evaluated in a biocontact oxidation reactor (BOR) and a sequencing batch reactor (SBR). During 3 months of continuous operation, the two bioreactors had similar treatment efficiencies (polyvinyl alcohol, PVA, 74.5-81.3%; COD, 73.5-77.4%; 2.15 pH decreases). Molecular biological analysis indicated that the microbial communities underwent dramatic changes during the operation, in which the SBR was superior to the BOR in retaining the alkali bacteria at the start-up stage, however, the BOR seemed to be more advantageous when the frequently changing influents were considered. The bacterial communities in BOR and SBR were diverse and included Bacteroidetes, Firmicutes, Proteobacteria, Actinobacteria and an unidentified cluster. Among these only Paracoccus sp. was successfully isolated and confirmed to have the ability to degrade PVA.

Influence of tetracycline exposure on the growth of wheat seedlings and the rhizosphere microbial community structure in hydroponic culture

In this study, the effects of tetracycline exposure on wheat growth and the microbial community structure in the rhizosphere were investigated under hydroponic culture conditions. Exposure to various concentrations of tetracycline resulted in significant suppression of the growth of wheat roots and shoots, with minimum doses of 0.8 mg L−1 and 4 mg L−1 resulting in inhibition rates of 32% and 15.4%, respectively. Complete inhibition of the growth of these two parts of wheat plants was observed in response to treatment with tetracycline at 20 mg L−1 and 100 mg L−1, respectively. However, the germination of wheat seeds was not sensitive to exposure to tetracycline. The effects of tetracycline exposure on the microbial community in the wheat rhizosphere were evaluated through traditional cultivation and molecular biological analyses. The cultivation results indicated that bacteria were the dominant population, being present in concentrations of 1× 108–2.45× 109CFUs mL−1, although 39% to 87% inhibition occurred in response to tetracycline. The concentration of fungi increased in all tetracycline treated samples to 2.5 to 15.8 times that of the control. The highest concentration of fungi (4.27× 108 CFU mL−1) was observed in response to 60 mg L−1 tetracycline after 15 days of cultivation. In this stage, a large amount of fungal colonies was observed on the surface of the culture solution, the wheat roots became rotted and the plants became atrophic or even died. Molecular biological analysis indicated that the bacterial community structure was significantly different in samples that were exposed to high levels of tetracycline (over 20 mg L−1) than in samples that were exposed to lower concentrations. As the concentration of tetracycline increased, the diversity of the bacteria decreased. Additionally, several dominant sensitive species such as Sphingobacterium multivorum were suppressed by tetracycline, while some resistant species such as Acinetobacter sp. appeared or were conserved. The bacteria population tended to stabilize when the drug concentration exceeded 40 mg L−1.

Effect of trace hydrazine addition on the functional bacterial community of a sequencing batch reactor performing completely autotrophic nitrogen removal over nitrite.

A sequencing batch reactor (SBR) was conducted to perform completely autotrophic nitrogen removal over nitrite (CANON). The effect of long-term trace N2H4 addition on ammonium oxidizing bacteria (AOB) and anaerobic AOB (AnAOB) in the CANON system was investigated. AOB and AnAOB primarily related to Nitrosococcus, Nitrosomonas and Candidatus scalindua, respectively. Before and after trace N2H4 addition, the estimates of AOB population decreased from 1.03×10(7) to 6.25×10(4)copies/g (dry sludge), but that of AnAOB increased from 3.14×10(9) to 5.86×10(10)copies/g (dry sludge). Despite there was a partially negative impact on AOB growth, the trace N2H4 addition exerted a stronger inhibition on nitrite oxidizing bacteria (NOB) and promoted AnAOB growth, which improved the nitrogen removal of the CANON system. Sludge granules enriched under long-term trace N2H4 addition were spherical and ellipsoidal, and the aerobic AOB were mainly located on the outer layers while AnAOB occupied most of the interior parts.

Influence of Chicken Manure Fertilization on Antibiotic-Resistant Bacteria in Soil and the Endophytic Bacteria of Pakchoi.

Animal manure is commonly used as fertilizer for agricultural crops worldwide, even though it is believed to contribute to the spread of antibiotic resistance from animal intestines to the soil environment. However, it is unclear whether and how there is any impact of manure fertilization on populations and community structure of antibiotic-resistant endophytic bacteria (AREB) in plant tissues. To investigate the effect of manure and organic fertilizer on endophytic bacterial communities, pot experiments were performed with pakchoi grown with the following treatments: (1) non-treated; (2) chicken manure-treated and (3) organic fertilizer-treated. Manure or organic fertilizer significantly increased the abundances of total cultivable endophytic bacteria (TCEB) and AREB in pakchoi, and the effect of chicken manure was greater than that of organic fertilizer. Further, 16S rDNA sequencing and the phylogenetic analysis indicated that chicken manure or organic fertilizer application increased the populations of multiple antibiotic-resistant bacteria (MARB) in soil and multiple antibiotic-resistant endophytic bacteria (MAREB) in pakchoi. The identical multiple antibiotic-resistant bacterial populations detected in chicken manure, manure- or organic fertilizer-amended soil and the vegetable endophytic system were Brevundimonas diminuta, Brachybacterium sp. and Bordetella sp., suggesting that MARB from manure could enter and colonize the vegetable tissues through manure fertilization. The fact that some human pathogens with multiple antibiotic resistance were detected in harvested vegetables after growing in manure-amended soil demonstrated a potential threat to human health.

Analysis of the bacterial community in a full-scale printing and dyeing wastewater treatment system based on T-RFLP and 454 pyrosequencing

In this study, the bacterial dynamics and structure compositions in the two-stage biological process of a full-scale printing and dyeing wastewater (PDW) treatment system were traced and analyzed by terminal restriction fragment length polymorphism (T-RFLP) and 454 pyrosequencing techniques. T-RFLP analysis showed that the microbial communities experienced significant variation in the process of seed sludge adaptation to the PDW environments and were in constant evolution during the whole running period of the system, despite the constant COD and color removal effects. Pyrosequencing results indicated that the two-stage biological system harbored rather diverse bacteria, with Proteobacteria being the predominant phylum during the steady running period, although its microbial compositions differed. The first-stage aerobic tank was dominated by α-Proteobacteria (89.05% of Proteobacteria), whereas in the second-stage aerobic tank, β- and γ-Proteobacteria, besides α-Proteobacteria, were the dominant bacterial populations.

Plant Growth, Antibiotic Uptake, and Prevalence of Antibiotic Resistance in an Endophytic System of Pakchoi under Antibiotic Exposure

Antibiotic contamination in agroecosystems may cause serious problems, such as the proliferation of various antibiotic resistant bacteria and the spreading of antibiotic resistance genes (ARGs) in the environment or even to human beings. However, it is unclear whether environmental antibiotics, antibiotic resistant bacteria, and ARGs can directly enter into, or occur in, the endophytic systems of plants exposed to pollutants. In this study, a hydroponic experiment exposing pakchoi (Brassica chinensis L.) to tetracycline, cephalexin, and sulfamethoxazole at 50% minimum inhibitory concentration (MIC) levels and MIC levels, respectively, was conducted to explore plant growth, antibiotic uptake, and the development of antibiotic resistance in endophytic systems. The three antibiotics promoted pakchoi growth at 50% MIC values. Target antibiotics at concentrations ranging from 6.9 to 48.1 µg·kg−1 were detected in the treated vegetables. Additionally, the rates of antibiotic-resistant endophytic bacteria to total cultivable endophytic bacteria significantly increased as the antibiotics accumulated in the plants. The detection and quantification of ARGs indicated that four types, tetX, blaCTX-M, and sul1 and sul2, which correspond to tetracycline, cephalexin, and sulfamethoxazole resistance, respectively, were present in the pakchoi endophytic system and increased with the antibiotic concentrations. The results highlight a potential risk of the development and spread of antibiotic resistance in vegetable endophytic systems.