Zhenyong Zhao

Fate of tetracycline, sulfonamide and fluoroquinolone resistance genes and the changes in bacterial diversity during composting of swine manure

This study monitored the abundance of antibiotic resistant genes (ARGs) and the bacterial diversity during composting of swine manure spiked with chlortetracycline, sulfadiazine and ciprofloxacin at two different levels and a control without antibiotics. Resistance genes of tetracycline (tetQ, tetW, tetC, tetG, tetZ and tetY), sulfonamide (sul1, sul2, dfrA1 and dfrA7) and fluoroquinolone (gyrA and parC) represented 0.02-1.91%, 0.67-10.28% and 0.00005-0.0002%, respectively, of the total 16S rDNA copies in the initial composting mass. After 28-42 days of composting, these ARGs, except parC, were undetectable in the composting mass indicating that composting is a potential method of manure management. Polymerase chain reaction-denaturing gradient gel electrophoresis analysis of bacterial 16S rDNA of the composting mass indicated that the addition of antibiotics up to 100, 20 and 20mg/kg of chlortetracycline, sulfadiazine and ciprofloxacin, respectively, elicited only a transient perturbation and the bacterial diversity was restored in due course of composting.

Composting of swine manure spiked with sulfadiazine, chlortetracycline and ciprofloxacin

The fate of chlortetracycline (CTC), sulfadiazine (SDZ) and ciprofloxacin (CIP) during composting of swine manure and their effect on composting process were investigated. Swine manure was spiked with antibiotics, mixed with saw dust (1:1 on DW basis) and composted for 56 d. Antibiotics were spiked to a final concentration of 50 mg/kg CTC+10 mg/kg SDZ+10 mg/kg CIP (High-level) or 5 mg/kg CTC+1 mg/kg SDZ+1 mg/kg CIP (Low-level), and a control without antibiotics. Antibiotics at high concentrations delayed the initial decomposition that also affected the nitrogen mineralization. CTC and SDZ were completely removed from the composting mass within 21 and 3d, respectively; whereas, 17-31% of the spiked CIP remained in the composting mass. Therefore, composting could effectively remove the CTC and SDZ spiked even at high concentrations, but the removal of ciprofloxacin (belonging to fluoroquinolone) needs to be improved, indicating this antibiotic may get into the ecosystem through land application of livestock compost.

Effects of rhamnolipids on cell surface hydrophobicity of PAH degrading bacteria and the biodegradation of phenanthrene.

The effects of rhamnolipids produced by Pseudomonas aeruginosa ATCC9027 on the cell surface hydrophobicity (CSH) and the biodegradation of phenanthrene by two thermophilic bacteria, Bacillus subtilis BUM and P. aeruginosa P-CG3, and mixed inoculation of these two strains were investigated. Rhamnolipids significantly reduced the CSH of the hydrophobic BUM and resulted in a noticeable lag period in the biodegradation. However, they significantly increased the CSH and enhanced the biodegradation for the hydrophilic P-CG3. In the absence of rhamnolipids, a mixed inoculation of BUM and P-CG3 removed 82.2% of phenanthrene within 30 days and the major contributor of the biodegradation was BUM (rapid degrader) while the growth of P-CG3 (slow degrader) was suppressed. Addition of rhamnolipids promoted the surfactant-mediated-uptake of phenanthrene by P-CG3 but inhibited the uptake through direct contact by BUM. This resulted in the domination of P-CG3 during the initial stage of biodegradation and enhanced the biodegradation to 92.7%.

Biosurfactants from Acinetobacter calcoaceticus BU03 enhance the solubility and biodegradation of phenanthrene

A thermophilic bacterial strain, Acinetobacter calcoaceticus BU03, with a biosurfactant‐producing capability, was isolated from petroleum‐contaminated soil with an improved procedure which employed the solubilization of polycyclic aromatic hydrocarnons (PAHs), i.e. naphthalene in agar plate, as a selection criterion. Crude biosurfactant was recovered from the culture of BU03 by extraction with n‐hexane, and its properties were investigated. Biosurfactants from A. calcoaceticus BU03 constitute a thermo‐stable mixture, composed of different agents with surface activities. At their critical micelle concentration (CMC) of 152.4 mg L−1, the crude biosurfactants produced from A. calcoaceticus BU03 decreased the air–water surface tension to 38.4 mN m−1. In thermophilic conditions, the emulsifying activity is 2.8 times that of Tween 80. The effects of the biosurfactants produced by A. calcoaceticus on the solubility and biodegradation of PAHs were investigated in batch systems. Biosurfactants produced by A. calcoaceticus BU03 at 25 times their CMC significantly increased the apparent aqueous solubility of phenanthrene (PHE), pyrene (PYR) and benzo(a)pyrene (B[a]P) to 54.3, 6.33 and 2.08mg L−1, respectively. In aqueous system, the biosurfactants at concentrations of 0.5 CMC and 1 CMC slightly enhanced the biodegradation of PHE by a consortium of PAH‐degrading microrganisms. Results indicate that biosurfactants from A. calcoaceticus BU03 have potential to enhance the removal of PAHs from contaminated sites.

Degradation of tetracycline and sulfadiazine during continuous thermophilic composting of pig manure and sawdust

During composting, the thermophilic phase resulted in high degradation of antibiotics in the composting mass; thus temperature is considered as the major factor for degradation of antibiotics. Therefore, to achieve complete removal of antibiotics, the effect of continuous thermophilic composting on the degradation of antibiotics and their effect on antibiotic resistant bacteria in the pig manure were evaluated. Pig manure was mixed with sawdust, spiked with tetracycline (10 and 100 mg/kg) and sulfadiazine (2 and 20 mg/kg) on dry weight (DW) basis and composted at 55°C for six weeks. Based on the organic decomposition, the antibiotics did not affect the composting process significantly, but negatively influenced the bacterial population. Tetracycline clearly exhibited a negative but marginal influence on carbon decomposition at 100 mg/kg level. The bacterial population initially decreased steeply ∼2 logs and slowly increased thereafter. Sulfadiazine and tetracycline resistant bacterial populations were stable/marginally increased after an initial decrease of about 2 or 3–5 logs, respectively. Sulfadiazine was not detectable after three days; whereas, ∼8% of tetracycline was detected after 42 days of composting with a t1/2 of ∼11 days, irrespective of the initial concentration. The presence of tetracycline in the compost after 42 days of thermophilic composting indicates the involvement of a mesophilic microbial-mediated degradation; however, further studies are required to confirm the direct microbial involvement in the degradation of antibiotics.

Synergistic effect of thermophilic temperature and biosurfactant produced by Acinetobacter calcoaceticus BU03 on the biodegradation of phenanthrene in bioslurry system.

This study aimed at investigating the synergistic effect of temperature and biosurfactant on the biodegradation of phenanthrene in bioslurry. Bench-scale bioslurry experiments were conducted at 25 and 55°C. The desorption rate coefficients of phenanthrene (K(des)) obtained using the pseudo-first order model were 0.0026 and 0.0035 kg mg(-1)h(-1) at 25 and 55°C, respectively. Addition of 1500 mg L(-1) biosurfactant, produced by Acinetobacter calcoaceticus BU03, marginally increased the K(des) at 25°C since most of biosurfactant was sorbed onto soil; however, significantly increased the K(des) to 0.0087 kg mg(-1)h(-1) at 55°C as the thermophilic temperature reduced the adsorption of the biosurfactant onto soil and subsequently enhanced the desorption of phenanthrene. The biodegradation of phenanthrene well fitted pseudo-first order kinetics based on the assumption that biodegradation was limited by the desorption. About 78.7% of phenanthrene was degraded in 30 days at 25°C; and addition of biosurfactant did not affect the biodegradation. However, addition of the biosurfactant or inoculation of A. calcoaceticus BU03 at 55°C significantly enhanced the biodegradation by increasing the K(des). Results indicate that synergistic application of thermophilic temperature and biosurfactant or inoculation of biosurfactant producing microorganisms is an effective and innovative method to enhance the efficiency of PAH degradation in bioslurry system.

Role of non‐ionic surfactants and plant oils on the solubilization of organochlorine pesticides by oil‐in‐water microemulsions

Screening low‐cost, high efficacy and environmentally safe surface active agents is critical for achieving successful surfactant‐enhanced remediation (SER) of soil contaminated with hydrophobic organic compounds. This study reports the solubilization of organochlorine pesticides (DDT or γ‐HCH) in oil‐in‐water (Winsor I) microemulsions (µE) composed of non‐ionic surfactant (Tween 80 or Triton X‐100), plant oil (linseed oil or soybean oil), and the cosurfactant (1‐pentanol). Results show that the cosurfactant to surfactant ratio (C/S ratio, w/w) is the major factor influencing the microemulsion formation, and C/S ratios of 1:3 and 1:6 are superior to 1:1 for microemulsion formation. 66.9–95.6% and 51.9–80.9% of DDT solubilization enhancements were achieved by microemulsions based respectively on Tween 80 or Triton X‐100 as compared to their respective surfactant solution alone, indicating the higher solubilizing capacities of microemulsion systems. The solubilization of γ‐HCH also increased by 40.6–57.5% in microemulsion formed with Tween 80 and 43.0–65.8% in microemulsion formed with Triton X‐100, compared with that in corresponding surfactant solutions only. Further studies revealed that both cosurfactant content and oil content could influence the solubilizing capacity of microemulsions system, and higher solubilizing capacity could be obtained when more cosurfactant or oil were emulsified in microemulsion system. Between the two, oil content is more influential than cosurfactant content. The present results affirm the effective role of microemulsions formed with Tween 80 and Triton X‐100 in enhancing the solubilization of DDT and γ‐HCH which would facilitate remediation of soils contaminated with these compounds.

Influence of different mixing ratios on in-vessel co-composting of sewage sludge with horse stable straw bedding waste: maturity and process evaluation

Composting sewage sludge alone would reduce the decomposition efficiency due to free limited porosity in sludge. To alleviate this, the use of horse stable straw bedding waste (HSB) was evaluated as a co-composting material with sewage sludge in a 10 tonnes day−1 in-vessel composter for a period of 7 days before curing in a static aeration pile. Sludge was mixed with HSB at 1 : 1.5 (HSL) and 1 : 2.9 (LSL) on a fresh weight basis. After a composting period of 56 days, both mixing ratios demonstrated to be feasible with LSL having a better organic decomposition and a shorter time to reach maturity. The overall decomposition rates were 52.0 and 58.9% (dry weight basis) for HSL and LSL, respectively. In both treatments, temperature in the in-vessel composters could reach 65°C, which was sufficient to remove the pathogens. Although both products were free of pathogens, HSL exhibited a higher ammoniacal nitrogen contents but a lower seed germination index than that of LSL indicating a higher phytotoxicity and a longer curing period would be required. It can be concluded that HSB provided a better composting conditions at a mixing ratio of 1 : 2.9

In-vessel co-composting of horse stable bedding waste and blood meal at different C/N ratios: process evaluation

Abattoir blood meal is rich in nitrogen and its potential as a co-composting material for horse stable bedding waste was evaluated at two C/N ratios – 32 (LBM, low blood meal) and 16 (HBM, high blood meal) – to improve the nutrient contents of the final compost. The mix was composted for 7 days in a 10 tonne/day in-vessel composter and cured aerobically. After 56 days of composting, the ammoniacal-N, CO 2 evolution rate and C/N ratio of both LBM and HBM were within the guideline values; however, delayed decomposition and lower seed germination index were observed with HBM. In addition, HBM resulted in 84% loss of the initial ammoniacal-N. Almost similar organic decompositions, 62.4% and 59.6% with LBM and HBM, respectively, were achieved. However, a stable compost product can be obtained within 6–7 weeks with LBM, whereas>8 weeks were required for HBM composting. Therefore, co-composting at the C/N ratio of 32 is recommended to achieve odour-free and faster composting.

Microemulsion-enhanced remediation of soils contaminated with organochlorine pesticides

Soil contaminated by organic pollutants, especially chlorinated aromatic compounds such as DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane), is an environmental concern because of the strong sorption of organochlorine pesticide onto the soil matrix and persistence in the environment. The remediation of organochlorine pesticide contaminated soils through microemulsion is an innovative technology to expedite this process. The remediation efficiency was evaluated by batch experiments through studying the desorption of DDT and hexachlorocyclohexane (γ-HCH) and sorption of microemulsion composed of Triton X-100, 1-pentanol and linseed oil in the soil–surfactant–water suspension system. The reduction of desorption efficiency caused by the sorption loss of microemulsion components onto the soil could be corrected by the appropriate adjustment of C/S (Cosurfactant/Surfactant) and O/S (Oil/Surfactant) ratio. The C/S and O/S ratios of 1:2 and 3:20 were suitable to desorb DDT and γ-HCH from the studied soils because of the lower sorption of Triton X-100 onto the soil. Inorganic salts added in microemulsion increased the pesticides desorption efficiency of pesticides and calcium chloride has a stronger ability to enhance the desorption of DDT than sodium chloride. From the remediation perspective, the balance of surfactant or cosurfactant sorbed to soil and desorption efficiency should be taken into consideration to enhance the remediation of soils contaminated by organochlorine pesticides.