Shailesh R. Dave

Isolation and characterization of Bacillus thuringiensis for Acid red 119 dye decolourisation

Studies were carried out to isolate Acid red 119 (AR-119) resistant and decolourising bacteria from dye contaminated soil and water samples. Six morphologically distinct bacterial isolates resistant to 100 ppm AR-119 dye were isolated directly from the soil and waste contaminated with azo dyes. The most efficient isolate, which showed decolourisation zone of 44 mm on 100 ppm AR-119 containing plate was identified as Bacillus thuringiensis SRDD. Gradual adaptation increased the efficiency of the isolate and within 7h of incubation it showed decolourisation up to 1000 ppm of AR-119 dye in liquid medium. Addition of 300 ppm of AR-119 in each step in ongoing dye decolourisation flask gave more than 90% decolourisation of 300 ppm AR-119 in time as short as 1.25 h. The developed B. thuringiensis showed 50-60% decolourisation of 5000 ppm AR-119 in 7d of incubation. This organism was also able to remove more than 98%, 92%, 95% and 95% colour of C.I. Acid brown 14, C.I. Acid black 210, C.I. Acid violet 90 and C.I. Acid yellow 42 azo dyes at 100 ppm concentration in 24h, respectively. When the developed isolate was studied for bioremediation of actual azo dye contaminated waste it removed 70% colour from the waste in 24h. The developed B. thuringiensis exhibited excellent resistance and decolourisation ability to AR-119 and other acid azo dyes.

Characterization of arsenic resistant and arsenopyrite oxidizing Acidithiobacillus ferrooxidans from Hutti gold leachate and effluents

Four arsenic resistant ferrous oxidizers were isolated from Hutti Gold Mine Ltd. (HGML) samples. Characterization of these isolates was done using conventional microbiological, biochemical and molecular methods. The ferrous oxidation rates with these isolates were 16, 48, 34 and 34 mg L(-1)h(-1) and 15, 47, 34 and 32 mg L(-1)h(-1) in absence and presence of 20 mM of arsenite (As3+) respectively. Except isolate HGM 8, other three isolates showed 2.9-6.3% inhibition due to the presence of 20 mM arsenite. Isolate HGM 8 was able to grow in presence of 14.7 g L(-1) of arsenite, with 25.77 mg L(-1)h(-1) ferrous oxidation rate. All the four isolates were able to oxidize iron and arsenopyrite from 20 g L(-1) and 40 g L(-1) refractory gold ore and 20 g L(-1) refractory gold concentrate. Once the growth was established pH adjustment was not needed inspite of ferrous oxidation, which could be due to concurrent oxidation of pyrite. Isolate HGM 8 showed the final cell count of as high as 1.12 x 10(8) cells mL(-1) in 40 g L(-1) refractory gold ore. The isolates were grouped into one haplotypes by amplified ribosomal DNA restriction analysis (ARDRA). The phylogenetic position of HGM 8 was determined by 16S rDNA sequencing. It was identified as Acidithiobacillus ferrooxidans and strain name was given as SRHGM 1.

Development of Leptospirillum ferriphilum dominated consortium for ferric iron regeneration and metal bioleaching under extreme stresses

Activated iron oxidizing consortium SR-BH-L enriched from Rajpardi lignite mine soil sample gave iron oxidation rate 1954 mg/L/h. Developed novel polystress resistant consortium oxidized ferrous iron under 11cP viscosity, 7.47 M ionic strength, 2.3 pH and g/L of 0.50 cadmium, 3.75 copper, 0.20 lead, 92.00 zinc, 6.4 sodium, 5.5 chloride, 154 sulphate and 393.8 TDS. The developed consortium showed 78.0% and 70.0% copper and zinc extraction from polymetallic bulk concentrate in monophasic bioleaching process. The bioregenerated ferric by the consortium in leachate showed 80.81% and 54.0% copper and zinc leaching in only 30 and 90 min. The DGGE analysis indicated the presence of 11 OTUs in the consortium. 16S rRNA gene sequence (JN797729) of the dominant band on DGGE shared >99% similarity with Leptospirillum ferriphilum. RE digestion analysis of the total 16S rRNA gene also illustrated the dominance of L. ferriphilum in the consortium.

Development of two-step process for enhanced biorecovery of Cu-Zn-Ni from computer printed circuit boards.

Metal pollution due to the huge electronic waste (E-waste) accumulation is widespread across the globe. Extraction of copper, zinc and nickel from computer printed circuit boards (c-PCB) with a two-step bleaching process using ferric sulphate generated by Leptospirillum ferriphilum dominated consortium and the factors influencing the process were investigated in the present study. The studied factors with 10 g/L pulp density showed that pH 2.0 was optimum which resulted in 87.50-97.80% Cu-Zn-Ni extraction. Pre-treatment of PCB powder with acidified distilled water and NaCl solution showed 3.80-7.98% increase in metal extraction corresponding to 94.08% Cu, 99.80% Zn and 97.99% Ni extraction. Particle size of 75 μm for Cu and Zn while 1680 μm for Ni showed 2-folds increase in metal extraction, giving 97.35-99.80% Cu-Zn-Ni extraction in 2-6 days of reaction time. Whereas; 2.76-3.12 folds increase in Cu and Zn extraction was observed with the addition of 0.1% chelating agents. When the studies were carried out with high pulp density, ferric iron concentration of 16.57 g/L was found to be optimum for metal extraction from 75 g/L c-PCB and c-PCB addition in multiple installments resulted in 8.81-26.35% increase in metal extraction compared to single addition. The studied factors can be implemented for the scale-up aimed at faster recovery of multimetals from E-waste and thereby providing a secondary source of metal in an eco-friendly manner.

Chemical and biological processes for multi-metal extraction from waste printed circuit boards of computers and mobile phones.

E-waste printed circuit boards (PCB) of computers, mobile-phones, televisions, LX (LongXiang) PCB in LED lights and bulbs, and tube-lights were crushed to ≥250 µm particle size and 16 different metals were analysed. A comparative study has been carried out to evaluate the extraction of Cu–Zn–Ni from computer printed circuit boards (c-PCB) and mobile-phone printed circuit boards (m-PCB) by chemical and biological methods. Chemical process showed the extraction of Cu–Zn–Ni by ferric sulphate was best among the studied chemical lixiviants. Bioleaching experiments were carried out with the iron oxidising consortium, which showed that when E-waste and inoculum were added simultaneously in the medium (one-step process); 60.33% and 87.50% Cu, 75.67% and 85.67% Zn and 71.09% and 81.87% Ni were extracted from 10 g L-1 of c-PCB and m-PCB, respectively, within 10–15 days of reaction time. Whereas, E-waste added after the complete oxidation of Fe2+ to Fe3+ iron containing medium (two-step process) showed 85.26% and 99.99% Cu, 96.75% and 99.49% Zn and 93.23% and 84.21% Ni extraction from c-PCB and m-PCB, respectively, only in 6–8 days. Influence of varying biogenerated Fe3+ and c-PCB concentrations showed that 16.5 g L-1 of Fe3+ iron was optimum up to 100 g L-1 of c-PCB. Changes in pH, acid consumed and redox potential during the process were also studied. The present study shows the ability of an eco-friendly process for the recovery of multi-metals from E-waste even at 100 g L-1 printed circuit boards concentration.

Selection of Leptospirillum ferrooxidans SRPCBL and development for enhanced ferric regeneration in stirred tank and airlift column reactor

Presence of Leptospirillum ferrooxidans plays significant role in ferric sulphate generation during bioleaching process. Thus, an attempt was made to select L. ferrooxidans from the polymetallic concentrate leachate and further developed it for enhanced ferric iron regeneration from the leachate in shake flask, stirred tank and column reactor. When ferric to ferrous iron ratio in the shake flask reached to 20:1, L. ferrooxidans out competed Acidithiobacillus ferrooxidans and accounted for more than 99% of the total population. The isolate was confirmed by 16S rRNA genes sequence analysis and named as L. ferrooxidans SRPCBL. When the culture was exposure to UV dose and the oxidation-reduction potential of the inoculation medium was adjusted to 40 0mV by ferrous:ferric iron ratio, the IOR reached to as high as 1.2 g/L/h in shake flask, even with initial ferrous iron concentration of 200 g/L. The chalcopyrite concentrate leachate containing 12.8, 15.7, and 42.0 g/L ferrous iron, ferric iron and copper, respectively was studied for ferric iron regeneration with the developed polymetallic resistant L. ferrooxidans SRPCBL in stirred tank and a developed biofilm airlift column, the highest IOR achieved were 2.20 g/L/h and 3.1 g/L/h, respectively, with ferrous oxidation efficiency of 98%. The ferric regeneration ability of the developed isolate from the leachate proves useful for a two-stage metal extraction process.

Bacterial Degradation of Azo Dye Containing Wastes

Till the late nineteenth century, all the dyes used were more or less natural with main sources like plants, insects and molluscs, and were mostly prepared on a small scale. It was only after 1856 that with Perkin’s historic discovery of the first synthetic dye, mauveine was manufactured on a large scale. At present there are more than 100,000 commercial dyes available with a estimated production of 7 × 105–1 × 106 tons per year (Robinson et al., Bioresour Technol 77:247–255, 2001). These dyes are used extensively in the paper, clothing, food, cosmetic and pharmaceutical industries. Because of the diversity of the dye components available for synthesis, a large number of structurally different synthetic dyes are today utilized for coloration. Among synthetic dyes, azo dyes are the largest and versatile class of dyes which account for more than 50 % of the dyes produced annually

Isolation, identification, characterization and polymetallic concentrate leaching studies of tryptic soy- and peptone-resistant thermotolerant Acidithiobacillus ferrooxidans SRDSM2.

Acidithiobacillus ferrooxidans strain SRDSM2 was isolated from silica containing soil sample collected at a Rajpardi lignite mine. The strain responded to the addition of 0.5 g/L peptone and 1.0 g/L tryptone soya broth in the ferrous sulphate tryptone soya broth (ITSB) medium with 35.3% and 29.6% increase in iron oxidation rate (IOR), but decrease in the IOR at higher peptone or tryptone soya broth levels. The presence of 4 mM of zinc as zinc sulphate in the medium increased the IOR by 24.4%. Forty percent of the inoculated cells survived even after exposure at 80 °C for 120 min and showed 30% ferrous iron oxidation. The Vmax and Ks for iron oxidation by the isolate were 344.82 mg/L/h and 32.25 g/L respectively. The isolate was able to oxidized ferrous iron even in presence of 4.06 M ionic strength of medium and leached>85% copper and zinc from the polymetallic concentrate. Thus, this isolate can be used for bioextraction of metals from polymetallic concentrate.

A novel biphasic leaching approach for the recovery of Cu and Zn from polymetallic bulk concentrate

In scale-up biphasic leaching process of polymetallic concentrate, the ferric bioregeneration cycles were performed in 15.0L down flow packed bed reactor; whereas the chemical leaching cycles were done using the biogenerated ferric in an indigenously designed 10.0L stirred tank reactor. The consortium took 25cycles for proper biofilm formation. It showed highest iron oxidation rate (IOR) of 3908.21mg/L/h at 25thcycle under no polymetallic stress. Even under stressed conditions, it was 2650-558mg/L/h. Cu extractions were 86.63-46.51 and Zn extractions were 67.89-14.74% in 1st-4thcycle, respectively. The developed consortium exhibited 17-51times higher IOR compared to original wild type consortium. Extraction isotherm for zinc with 30% Cyanex® 301 indicated that a total of two stages are required for its complete extraction using the phase ratio of 2:1 at equilibrium pH 1.5, leaving behind Fe(II) in the raffinate.

Pretreatment optimization of Sorghum pioneer biomass for bioethanol production and its scale-up.

Based on one parameter at a time, saccharification of delignified sorghum biomass by 4% and 70% v/v sulfuric acid resulted in maximum 30.8 and 33.8 g% sugar production from biomass respectively. The Box Behnken Design was applied for further optimization of acid hydrolysis. As a result of the designed experiment 36.3g% sugar production was achieved when 3% v/v H2SO4 treatment given for 60 min at 180°C. The process was scaled-up to treat 2 kg of biomass. During the screening of yeast cultures, isolate C, MK-I and N were found to be potent ethanol producers from sorghum hydrolyzate. Culture MK-I was the best so used for scale up of ethanol production up to 25 L capacity, which gave a yield of 0.49 g ethanol/g sugar from hydrolyzate obtained from 2 kg of sorghum biomass.