Zafar M. Khalid

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.

Antioxidant and antimicrobial attributes and phenolics of different solvent extracts from leaves, flowers and bark of Gold Mohar [Delonix regia (Bojer ex Hook.) Raf].

This paper describes the antioxidant and antimicrobial activities and phenolic components of different solvent (absolute methanol, absolute ethanol, absolute acetone, 80% methanol, 80% ethanol, 80% acetone and deionized water) extracts of leaves, flowers and bark of Gold Mohar [Delonix regia (Bojer ex Hook.) Raf.]. The extract yields from leaves, flowers and bark ranged from 10.19 to 36.24, 12.97 to 48.47 and 4.22 to 8.48 g/100 g dry weight (DW), respectively. Overall, 80% methanol extract produced from the leaves exhibited significantly (P < 0.05) higher antioxidant activity, with high phenolic contents (3.63 g GAE/100 g DW), total flavonoid contents (1.19 g CE/100 g DW), inhibition of peroxidation (85.54%), DPPH scavenging capacity (IC50 value 8.89 μg/mL) and reducing power (1.87). Similarly, this 80% methanol leaves extract also showed superior antimicrobial activity. HPLC analysis of the 80% methanol extracts for individual phenolics revealed the presence of gallic, protocatechuic and salicylic acid in leaves; gallic, protocatechuic, salicylic, trans-cinnamic and chlorogenic acid in flowers, and gallic acid in bark as the main (amount > 1.50 mg/100 g DW) phenolic acids. Besides, small amounts (<1.50 mg/100 g DW) of some other phenolic acids such as sorbic, sinapic, p-coumaric, m-coumaric, ferulic, caffeic, 3-hydroxybenzoic, 4-hydroxycinnamic and 4-hydroxybenzoic acids were also detected. The extracts of the tested parts of Gold Mohar, especially, the leaves, might be valuable for functional food and therapeutic applications.

Protein-mediated synthesis, pH-induced reversible agglomeration, toxicity and cellular interaction of silver nanoparticles

Casein, a milk protein, is used to produce biotolerable and highly stable silver nanoparticles with a fair control over their size without using any additional reducing agent. These silver nanoparticles undergo reversible agglomeration to form protein-silver nanoparticle composite agglomerates as pH approaches to the isoelectric point of casein protein (pI=4.6). These agglomerates can then easily be re-dispersed in alkaline aqueous media with no obvious change in their optical properties. The nanoparticles can withstand high salt concentration (~0.5M), and can also be freeze-dried, stored as dry powder and then dispersed in aqueous media whenever required. More interestingly, by controlling the concentration of casein protein and pH, it was also possible to control the self-assembly of silver nanoparticles to produce fairly uniform spherical agglomerates. The nanoparticles and their agglomerates were thoroughly characterized using UV-visible and FTIR spectroscopy, TEM, SEM and DLS, etc. Cytotoxicity of the hybrid materials was examined using a Resazurin based cytotoxicity assay. After determining the LD(50) using NIH/3T3 fibroblast cells, the cellular interaction of these hybrid nanoparticles was studied to examine the behavior of casein-coated nanoparticles for their potential bio-applications.

DNA damage in Pakistani agricultural workers exposed to mixture of pesticides.

A cross‐sectional study was designed to determine whether occupational exposure to a complex mixture of pesticides results in a significant increase of DNA damage in farmers chronically exposed to pesticides in open fields. Leukocytes from 47 agriculture workers exposed to pesticides and 50 controls were evaluated with comet assay. Workers recruitment was based on their exposure to pesticides during the spraying season on cotton crop. Serum from these individuals was also analyzed for pesticides presence using high performance liquid chromatography. Statistically significant difference (P < 0.001) in DNA damage of exposed individuals (mean ± S.D 14.80 ± 3.04 μm) was observed when compared with control group (6.54 ± 1.73 μm) as studied on the basis of comet tail length. Smokers had significantly higher mean comet tail length than nonsmokers and ex‐smokers in both workers (20.26 ± 3.53 vs. 14.19 ± 4.25, P < 0.001) and controls (7.86 ± 1.09 vs. 5.80 ± 1.59, P < 0.001), whereas age had a minimal effect on DNA damage (P < 0.05). The length of pesticide exposure is positively associated with DNA damage in exposed individuals (P < 0.001). Our study shows that chronic exposure to pesticides produces DNA damage in pesticide sprayers and suggests that this type of monitoring is recommended in preventive policies for pesticide sprayers. Environ. Mol. Mutagen., 2009.

Characterization of rhamnolipids produced by a Pseudomonas aeruginosa mutant strain grown on waste oils

Pseudomonas aeruginosa EBN-8 mutant rhamnolipids produced on waste oils were investigated using normal-phase thin layer chromatography and fast atom bombardment mass spectrometry. Negative ion mode mass spectra yielded [M – H]− ions and their fragment ions, which gave some indications on the sequence of rhamnolipid biosynthesis. Five rhamnolipid homologs [viz. RC10C10 (m/z 503), RC12C10 or RC10C12 (531), RRC10C8 or RRC8C10 (621), RRC10C10 (649) and RRC12C10 or RRC10C12 (677)] were detected in four rhamnolipid combinations under the different carbon sources. The prevalence of rhamnolipids was confirmed by Fourier transform infrared and one-dimensional proton nuclear magnetic resonance. We also observed some correlations between the tensioactive characteristics and structural chemistry of the rhamnolipid surfactants.

Physicochemical and surface-active properties of biosurfactant produced using molasses by a Pseudomonas aeruginosa mutant

Production of a microbial surfactant was studied by growing Pseudomonas aeruginosa EBN-8 mutant on varying concentrations (on the basis of total sugars) of clarified blackstrap molasses as a sole carbon and energy source with or without auxiliary synthetic nitrogen source in 250 mL shake flasks. The progress of fermentation process was monitored by measuring the production of metabolites, and surface-active and emulsification properties of the cell-free culture broth. The biosurfactant was isolated from the supernatant by acid precipitation followed by solvent extraction. The amount of rhamnolipids produced was determined by the orcinol method. The highest dry cell biomass (1.67 g/L) and rhamnolipid (1.45 g/L) yields were observed, at 96 h of incubation on 2% total sugars-based molasses amended with sodium nitrate (at C:N, 20:1) with the product yield related to dry cell biomass (Y P/X, g/g) of 0.869, specific product formation rate (V, h−1) of 0.295 and volumetric productivity rate (P V, g/L/h) of 0.015. The surface tension of this culture medium dropped to 28.0 from 50.0 mN/m.

Synthesis and use of self-assembled rhamnolipid microtubules as templates for gold nanoparticles assembly to form gold microstructures.

Natural unmodified rhamnolipids are thermally self-assembled into soft microtubules, which can produce gold nanoparticles onto themselves due to the presence of rhamnose sugar moieties at their surface. The loading of gold nanoparticles on composite microtubules can be controlled by varying the concentration of gold salt to rhamnolipid and the reaction temperature. The composite rhamnolipid-gold nanoparticle microtubules are then heat treated to produce porous gold microwire-like structures with fairly controlled nanostructured features, which may have interesting applications in catalysis, biosensing and electronics.

Paper and board mill effluent treatment with the combined biological-coagulation-filtration pilot scale reactor.

Pilot scale reactor based on combined biological-coagulation-filtration treatments was designed and evaluated for the treatment of effluent from a paper and board mill. Biological treatment by fed batch reactor (FBR) followed by coagulation and sand filtration (SF) resulted in a total COD and BOD reduction of 93% and 96.5%, respectively. A significant reduction in both COD (90%) and BOD (92%) was also observed by sequencing batch reactor (SBR) process followed by coagulation and filtration. Untreated effluent was found to be toxic, whereas the treated effluents by either of the above two processes were found to be non-toxic when exposed to the fish for 72h. The resultant effluent from FBR-coagulation-sand filtration system meets National Environmental Quality Standards (NEQS) of Pakistan and can be discharged into the environment without any risks.

Production of biosurfactant using different hydrocarbons by Pseudomonas aeruginosa EBN-8 mutant.

Abstract The present investigation dealt with the use of previously isolated and studied gamma-ray mutant strain Pseudomonas aeruginosa EBN-8 for the production of biosurfactant by using different hydrocarbon substrates viz. n-hexadecane, paraffin oil and kerosene oil, provided in minimal medium, as the sole carbon and energy sources. The batch experiments were conducted in 250 mL Erlenmeyer flasks, containing 50 mL minimal salt media supplemented with 1% (w/v) hydrocarbon substrate, inoculated by EBN-8 and incubated at 37 °C and 100 rpm in an orbital shaker. The sampling was done on 24 h basis for 10 d. The surface tension of cell-free culture broth decreased from 53 to 29 mN/m after 3 and 4 d of incubation when the carbon sources were paraffin oil and n-hexadecane, respectively. The largest reduction in interfacial tension from 26 to 0.4 mN/m was observed with n-hexadecane, while critical micelle dilution was obtained as 50 X CMC for paraffin oil as carbon source. When grown on n-hexadecane and paraffin oil, the EBN-8 mutant strain gave 4.1 and 6.3 g of the rhamnolipids/ L, respectively. These surface-active substances subsequently allowed the hydrocarbon substrates to disperse readily as emulsion in aqueous phase

Polyhexamethylene biguanide functionalized cationic silver nanoparticles for enhanced antimicrobial activity

Polyhexamethylene biguanide (PHMB), a broad spectrum disinfectant against many pathogens, was used as a stabilizing ligand for the synthesis of fairly uniform silver nanoparticles. The particles formed were characterized using UV-visible spectroscopy, FTIR, dynamic light scattering, electrophoretic mobility, and TEM to measure their morphology and surface chemistry. PHMB-functionalized silver nanoparticles were then evaluated for their antimicrobial activity against a gram-negative bacterial strain, Escherichia coli. These silver nanoparticles were found to have about 100 times higher bacteriostatic and bactericidal activities, compared to the previous reports, due to the combined antibacterial effect of silver nanoparticles and PHMB. In addition to other applications, PHMB-functionalized silver nanoparticles would be extremely useful in textile industry due to the strong interaction of PHMB with cellulose fabrics.