Response Surface Methodology for Optimization of Operational Parameters To Remove Ciprofloxacin from Contaminated Water in the Presence of a Bacterial Consortium.

Ciprofloxacin (CFX) is a broad-spectrum fluoroquinolone antibiotic that is widely used to treat bacterial infections in humans and other animals. However, its unwanted occurrence in any (eco)system can affect nontarget bacterial communities, which may also impair the performance of the natural or artificially established bioremediation system. The problem could be minimized by optimization of operational parameters via modeling of multifactorial tests. To this end, we used a Box-Behnken design in response surface methodology (RSM) to generate the experimental layout for testing the effect of the CFX biodegradation for four important parameters, that is, temperature (°C), pH, inoculum size (v/v %), and CFX concentration (mg L-1). For inoculation, a consortium of three bacterial strains, namely, Acenitobacter lwofii ACRH76, Bacillus pumilus C2A1, and Mesorihizobium sp. HN3 was used to degrade 26 mg L-1 of CFX. We found maximum degradation of CFX (98.97%; initial concentration of 25 mg L-1) at 2% inoculum size, 7 pH, and 35 °C of temperature in 16 days. However, minimum degradation of CFX (48%; initial concentration of 50 mg L-1) was found at pH 6, temperature 30 °C, and inoculum size 1%. Among different tested parameters, pH appears to be the main limiting factor for CFX degradation. Independent factors attributed 89.37% of variation toward CFX degradation as revealed by the value of the determination coefficient, that is, R 2 = 0.8937. These results were used to formulate a mathematical model in which the computational data strongly correlated with the experimental results. This study showcases the importance of parameter optimization via RSM for any bioremediation studies particularly for antibiotics in an economical, harmless, and eco-friendly manner.

A Patient with SIADH, Urinary Retention, Constipation, and Bell's Palsy following a Tick Bite.

Introduction. Lyme disease is the most common vector borne disease in the USA caused by the bacterium Borrelia burgdorferi. If untreated, Lyme disease can cause a variety of secondary symptoms often difficult to interpret. Some of the rare manifestations of Lyme disease include SIADH-like syndrome, enteroparesis, and urinary retention. Case Report. A 69-year-old male presented with anorexia and constipation and was found to have hyponatremia. Several days after admission, Bell's palsy developed and he experienced urinary retention requiring catheterization. Lyme disease was confirmed on serology, and he recalled a rash on his elbow four weeks prior. Ceftriaxone was started and the patient improved; he had multiple bowel movements after receiving laxatives and the Foley catheter was removed; serum sodium normalized with fluid restriction. He was discharged home and was well with symptoms completely resolved at three-month follow-up. Discussion. There should be a high alert of atypical presentation of this common tick bite associated infection. Review of the literature revealed ten similar cases, but only three of these patients were reported to have a combination of SIADH, urinary retention, and enteroparesis.

Biodegradation and Subsequent Toxicity Reduction of Co-contaminants Tribenuron Methyl and Metsulfuron Methyl by a Bacterial Consortium B2R.

AllyMax is a widely used herbicide formulation in wheat-rice cropping areas of the world. The residues of its active ingredients, tribenuron methyl (TBM) and metsulfuron methyl (MET), persist in soil and water as co-contaminants, and cause serious threats to nontarget organisms. This study was performed to assess the potential of a bacterial consortium for the degradation and detoxification of TBM and MET individually and as co-contaminants. A bacterial consortium (B2R), comprising Bacillus cereus SU-1, Bacillus velezensis OS-2, and Rhodococcus rhodochrous AQ1, capable of degrading TBM and MET in liquid cultures was developed. Biodegradation of TBM and MET was optimized using the Taguchi design of experiment. Optimum degradation of both TBM and MET was obtained at pH 7 and 37 °C. Regarding media composition, optimum degradation of TBM and MET was obtained in minimal salt medium (MSM) supplemented with glucose, and MSM without glucose, respectively. The consortium simultaneously degraded TBM and MET (94.8 and 80.4%, respectively) in cultures containing the formulation AllyMax, where TBM and MET existed as co-contaminants at 2.5 mg/L each. Mass spectrometry analysis confirmed that during biodegradation, TBM and MET were metabolized into simpler compounds. Onion (Allium cepa) root inhibition and Comet assays revealed that the bacterial consortium B2R detoxified TBM and MET separately and as co-contaminants. The consortium B2R can potentially be used for the remediation of soil and water co-contaminated with TBM and MET.

Operational parameters optimization for remediation of crude oil-polluted water in floating treatment wetlands using response surface methodology.

The application of floating treatment wetlands (FTWs) is an innovative nature-based solution for the remediation of polluted water. The rational improvement of water treatment via FTWs is typically based on multifactorial experiments which are labor-intensive and time-consuming. Here, we used the response surface methodology (RSM) for the optimization of FTW's operational parameters for the remediation of water polluted by crude oil. The central composite design (CCD) of RSM was used to generate the experimental layout for testing the effect of the variables hydrocarbon, nutrient, and surfactant concentrations, aeration, and retention time on the hydrocarbon removal in 50 different FTW test systems planted with the common reed, Phragmites australis. The results from these FTW were used to formulate a mathematical model in which the computational data strongly correlated with the experimental results. The operational parameters were further optimized via modeling prediction plus experimental validation in test FTW systems. In the FTW with optimized parameters, there was a 95% attenuation of the hydrocarbon concentration, which was very close to the 98% attenuation predicted by the model. The cost-effectiveness ratio showed a reduction of the treatment cost up to $0.048/liter of wastewater. The approach showed that RSM is a useful strategy for designing FTW experiments and optimizing operational parameters.

Immobilization of metribuzin-degrading bacteria on biochar: Enhanced soil remediation and bacterial community restoration.

Metribuzin (MB), a triazinone herbicide is extensively sprayed for weed control in agriculture, has been reported to contaminate soil, groundwater, and surface waters. In soil, MB residues can negatively affect not only the germination of subsequent crops but also disturb soil bacterial community. The present study describes the use of biochar as a carrier material to immobilize MB-degrading bacterial consortium, for remediation of MB-contaminated soil and restoration of soil bacterial community in soil microcosms. The bacterial consortium (MB3R) comprised four bacterial strains, i.e., Rhodococcus rhodochrous AQ1, Bacillus tequilensis AQ2, Bacillus aryabhattai AQ3, and Bacillus safensis AQ4. Significantly higher MB remediation was observed in soil augmented with bacterial consortium immobilized on biochar compared to the soil augmented with un-immobilized bacterial consortium. Immobilization of MB3R on biochar resulted in higher MB degradation rate (0.017 Kd-1) and reduced half-life (40 days) compared to 0.010 Kd-1 degradation rate and 68 day half-life in treatments where un-immobilized bacterial consortium was employed. It is worth mentioning that the MB degradation products metribuzin-desamino (DA), metribuzin-diketo (DK), and metribuzin desamino-diketo (DADK) were detected in the treatments where MB3R was inoculated either alone or in combination with biochar. MB contamination significantly altered the composition of soil bacteria. However, soil bacterial community was conserved in response to augmentation with MB3R immobilized on biochar. Immobilization of the bacterial consortium MB3R on biochar can potentially be exploited for remediation of MB-contaminated soil and protecting its microbiota.

Immobilization of metribuzin degrading bacterial consortium MB3R on biochar enhances bioremediation of potato vegetated soil and restores bacterial community structure.

Metribuzin (MB) is a triazinone herbicide used for the eradication of weeds in agriculture. Presence of its residues in agricultural soil can potentially harm the establishment of subsequent crops and structure of soil microbial populations. In this study, remediation potential of an MB degrading bacterial consortium MB3R immobilized on biochar was evaluated in potato vegetated soil. In potato vegetated soil augmented with MB3R alone and MB3R immobilized on biochar, 82 and 96% MB degradation was recorded respectively as compared to only 29.3% in un-augmented soil. Kinetic parameters revealed that MB3R immobilized biochar is highly proficient as indicated by significant increase in the rate of biodegradation and decrease in half-life of MB. Enhanced plant growth was observed when augmented with bacterial consortium either alone or immobilized on biochar. Presence of herbicide negatively affected the soil bacterial community structure. However, MB3R immobilized on biochar proved to be helpful for restoration of soil bacterial community structure affected by MB. This is the very first report that reveals improved remediation of contaminated soil and restoration of soil bacterial populations by use of the MB degrading bacterial consortium immobilized on biochar.

Application of a novel bacterial consortium BDAM for bioremediation of bispyribac sodium in wheat vegetated soil.

Plant-bacterial mutualism has tremendous potential for remediation of herbicide contaminated soils. Generally, bacterial inoculation helps plants to grow well in the contaminated environment. Here, we investigated the impact of bispyribac sodium (BS) degrading bacterial consortium (BDAM) on BS remediation, plant growth promotion and BS accumulation in plant parts. Wheat (Triticum aestivum) was planted in BS spiked soil and inoculated with BDAM. Inoculation showed a beneficial effect on plant biomass production and degradation of BS in the rhizosphere and the rhizosheath. After 40 and 60 days of inoculation, the degradation of BS was more than 96% and approximately 100% respectively in the planted and inoculated soil spiked with 2 and 5 mg kg-1 BS. However, in planted and un-inoculated soil, the degradation of BS was 72% after 60 days of sowing. Furthermore, inoculated bacterial strains colonized both in rhizo- and endosphere of the inoculated plants. In comparison with the un-inoculated soil, significantly less accumulation of BS was found in the roots and shoots of the plants growing in inoculated soil. We report the efficiency of plant-bacterial partnership for enhanced biodegradation of BS and to eliminate the BS residual toxicity to non-target plants.

Optimizing the metribuzin degrading potential of a novel bacterial consortium based on Taguchi design of experiment.

Metribuzin (MB) is used for control of weeds in crops like potato, maize and sugarcane. Its extensive and unjudicial use has resulted in various environmental issues; hence it is very critical to remediate this herbicide at the respective point source. Plant associated, MB degrading bacterial strains, Rhodococcus rhodochrous sp. AQ1, Bacillus tequilensis sp. AQ2, Bacillus aryabhattai sp. AQ3 and Bacillus safensis sp. AQ4 were isolated, and a consortium MB3R was developed. For degradation of MB by the consortium MB3R, various parameters i.e., pH, temperature, inoculum density and pesticide concentration were optimized by using Taguchi design of experiment (DOE). MB degradation was dependent upon all the four factors. The contribution of each factor on MB degradation was according to the order: temperature > inoculum density > pH > pesticide concentration. Fitness of Taguchi DOE in forecasting the optimum response, was confirmed experimentally by using optimized levels of the four factors i.e., pH 7.0, temperature 30 °C, pesticide concentration 45 mg l-1 and an inoculum density of 5.0 × 10 5 CFU ml-1 whereby 98.63% MB degradation was observed. Appearance and subsequent degradation of three MB metabolites, desamino-metribuzin (DA), diketo-metribuzin (DK) and desamino-diketo-metribuzin (DADK) during biodegradation by the consortium was observed.

Biodegradation of bispyribac sodium by a novel bacterial consortium BDAM: Optimization of degradation conditions using response surface methodology.

Bispyribac sodium (BS), is a selective, systemic and post emergent herbicide used to eradicate grasses and broad leaf weeds. Extensive use of this herbicide has engendered serious environmental concerns. Hence it is important to develop strategies for bioremediation of BS in a cost effective and environment friendly way. In this study a bacterial consortium named BDAM, comprising three novel isolates Achromobacter xylosoxidans (BD1), Achromobacter pulmonis (BA2), and Ochrobactrum intermedium (BM2), was developed by virtue of its potential for degradation of BS. Different culture conditions (temperature, pH and inoculum size) were optimized for degradation of BS by the consortium BDAM and the mutual interactions of these parameters were analysed using a 23 full factorial central composite design (CCD) based on Response Surface Methodology (RSM). The optimal values for temperature, pH and inoculum size were found to be 40 °C, 8 and 0.4 g/L respectively to achieve maximum degradation of BS (85.6%). Moreover, the interactive effects of these parameters were investigated using three dimensional surface plots in terms of maximum fitness function. Importantly, it was concluded that the newly developed consortium is a potential candidate for biodegradation of BS in a safe, cost-effective and environmentally friendly manner.

Identification and analysis of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from glyphosate-resistant Ochrobactrum intermedium Sq20.

BACKGROUND: Glyphosate is a herbicide that acts by inhibition of the enzyme, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), involved in the catalysis of an essential step in the biosynthesis of aromatic amino acids. The objective of this study was the isolation of glyphosate-resistant bacterial strains and subsequent characterization of the gene(s) encoding glyphosate resistance in these isolates. Using an enrichment culture technique, a glyphosate-resistant bacterium, Ochrobactrum intermedium Sq20 was isolated from glyphosate-contaminated indigenous soil and characterized. RESULTS: An open reading frame (ORF) comprising of 1353 bp potentially encoding aroAO. intermediumSq20 was amplified from O. intermedium Sq20. It showed 97% homology with aroA genes from other Ochrobactrum spp. Physicochemical characterization revealed that aroAO. intermediumSq20 encodes a polypeptide of 450 amino acids with a calculated molecular mass of 48.9782 kDa and an isoelectric point of 5.21. Secondary structure prediction of AroAO. intermediumSq20 demonstrated a high percentage of random coils and α helices. Methodical optimization and validation of the protein structure helped to build a reliable protein model indicating the presence of 91.8% amino acid residues in most favoured regions. In addition, strain Sq20 was found to be capable of complete degradation of glyphosate at 500 mg L-1 initial concentration as the sole carbon and energy source within 4 days. CONCLUSION: A glyphosate-resistant bacterial strain O. intermedium Sq20 was discovered. Sequence analysis and structure modelling demonstrated that AroAO. intermediumSq20 closely resembles class II EPSPS and possesses high glyphosate resistance. This provides a good foundation for functional analysis of experimentally derived crystal structures. The cloning and characterization of AroAO. intermediumSq20 will further help in understanding its role at the molecular level and its potential use in the production of glyphosate-resistant transgenic crops. © 2017 Society of Chemical Industry.

Enhanced degradation of textile effluent in constructed wetland system using Typha domingensis and textile effluent-degrading endophytic bacteria.

Textile effluent is one of the main contributors of water pollution and it adversely affects fauna and flora. Constructed wetland is a promising approach to remediate the industrial effluent. The detoxification of industrial effluent in a constructed wetland system may be enhanced by applying beneficial bacteria that are able to degrade contaminants present in industrial effluent. The aim of this study was to evaluate the influence of inoculation of textile effluent-degrading endophytic bacteria on the detoxification of textile effluent in a vertical flow constructed wetland reactor. A wetland plant, Typha domingensis, was vegetated in reactor and inoculated with two endophytic bacterial strains, Microbacterium arborescens TYSI04 and Bacillus pumilus PIRI30. These strains possessed textile effluent-degrading and plant growth-promoting activities. Results indicated that bacterial inoculation improved plant growth, textile effluent degradation and mutagenicity reduction and were correlated with the population of textile effluent-degrading bacteria in the rhizosphere and endosphere of T. domingensis. Bacterial inoculation enhanced textile effluent-degrading bacterial population in rhizosphere, root and shoot of T. domingensis. Significant reductions in COD (79%), BOD (77%) TDS (59%) and TSS (27%) were observed by the combined use of plants and bacteria within 72 h. The resultant effluent meets the wastewater discharge standards of Pakistan and can be discharged into the environment without any risks. This study revealed that the combined use of plant and endophytic bacteria is one of the approaches to enhance textile effluent degradation in a constructed wetland system.

Enhanced remediation of chlorpyrifos from soil using ryegrass (Lollium multiflorum) and chlorpyrifos-degrading bacterium Bacillus pumilus C2A1.

The combined use of plants and associated microorganisms has great potential for remediating soil contaminated with organic compounds such as pesticides. The objective of this study was to determine whether the bacterial inoculation influences plant growth promotion and chlorpyrifos (CP) degradation and accumulation in different parts of the plant. Ryegrass was grown in soil spiked with CP and inoculated with a pesticide degrading bacterial strain Bacillus pumilus C2A1. Inoculation generally had a beneficial effect on CP degradation and plant biomass production, highest CP degradation (97%) was observed after 45 days of inoculation. Furthermore, inoculated strain efficiently colonized in the rhizosphere of inoculated plant and enhanced CP and its primary metabolite 3,5,6-trichloro-2-pyridinol (TCP) degradation. There was significantly less CP accumulation in roots and shoots of inoculated plants as compared to uninoculated plants. The results show the effectiveness of inoculated exogenous bacteria to boost the remediation of CP contaminated sites and decrease levels of toxic pesticide residues in crop plants.

Biodegradation of chlorpyrifos and its hydrolysis product 3,5,6-trichloro-2-pyridinol by Bacillus pumilus strain C2A1.

A bacterial strain C2A1 isolated from soil was found highly effective in degrading chlorpyrifos and its first hydrolysis metabolite 3,5,6-trichloro-2-pyridinol (TCP). On the basis of morphology, physiological characteristics, biochemical tests and 16S rRNA sequence analysis, strain C2A1 was identified as Bacillus pumilus. Role of strain C2A1 in the degradation of chlorpyrifos was examined under different culture conditions like pH, inoculum density, presence of added carbon/nutrient sources and pesticide concentration. Chlorpyrifos was utilized by strain C2A1 as the sole source of carbon and energy as well as it was co-metabolized in the presence of glucose, yeast extract and nutrient broth. Maximum pesticide degradation was observed at high pH (8.5) and high inoculum density when chlorpyrifos was used as the sole source and energy. In the presence of other nutrients, chlorpyrifos degradation was enhanced probably due to high growth on easily metabolizable compounds which in turn increased degradation. The strain C2A1 showed 90% degradation of TCP (300 mg L(-1)) within 8 days of incubation.