BIOREMEDIATION OF HYDROCARBONS AND XENOBIOTICS

by DR. S.KARPAGAM Associate Professor of Botany QUEEN MARYS COLLEGE

3 Es

ECONOMY ENERGY EFFICIENCY

Water molecule
Dihydrogen Monoxide DHMO Dihydrogen Oxide, Hydrogen Hydroxide, Hydronium Hydroxide, Hydric acid.

water
Main source of fresh water

RAIN

On 14 November 2008, the Moon Impact Probe separated from the Chandrayaan struck the south pole to place its flag on the Moon. The probe impacted near Shackleton Crater at 20:31 ejecting underground soil that could be analysed for the presence of lunar water ice.

The estimated cost for the project was 386 crore (US$80 million).

The Cassini spacecraft also carries a set of sensitive magnetometers, a Visual and Infrared Mapping Spectrometer (VIMS) that will enable scientists to detect subtle changes in the brightness of surfaces in specific colours which are signatures of particular materials such as methane or water ice ESTIMATED COST 3.5 Million dollars

ANCIENT CIVILIZATIONS

GREAT BARRIER REEF

New islands emerge from coral reefs

Marine ecosystem is unique not only by its vast area but also by its influence on the climatic conditions of the earth as a whole, and weather conditions of regional areas. Sewage from cities, towns, toxic heavy metals, pesticides, hazardous chemicals, petroleum hydrocarbons are opened into the sea.

Marine ecosystem
Species diversity Unexplored regions Thermal vents Carbon sink

MARINE POLLUTION
Sewage from cities Industrial effluents Oil slick Pollution from warships Military activities

Oil spill

A xenobiotic is a chemical which is found in an organism but which is not normally produced or expected to be present in it. It can also cover substances which are present in much higher concentrations than are usual.

However, the term xenobiotics is very often used in the context of pollutants such as dioxins and polychlorinated biphenyls and their effect on the biota, because xenobiotics are understood as substances foreign to an entire biological system, i.e. artificial substances, which did not exist in nature before their synthesis by humans.

Inorganic water pollutants include:

*Acidity caused by industrial discharges (especially sulfur dioxide from power plants) *Ammonia from food processing waste *Chemical waste as industrial by-products *Fertilizers containing nitrates and *phosphates--which are found in stormwater runoff from agriculture *Heavy metals from motor vehicles (via urban stormwater runoff) and acid mine drainage

*Silt (sediment) in runoff from construction sites, logging, slash and burn practices or land clearing sites

Organic water pollutants include: *Detergents

*Disinfection by-products found in chemically disinfected drinking water, such as chloroform *Food processing waste, which can include oxygen-demanding substances, grease *Insecticides and herbicides, a huge range of organohalides and other chemical compounds
Contd

*Petroleum hydrocarbons, including fuels (gasoline, diesel fuel, jet fuels, and fuel oil) and lubricants (motor oil), and fuel combustion byproducts,. *Volatile organic compounds (VOCs), such as industrial solvents, from improper storage. *Chlorinated solvents, which are dense non-aqueous phase liquids (DNAPLs), may fall to the bottom of reservoirs, since they don't mix well with water and are denser. *Various chemical compounds found in personal hygiene and cosmetic products

CHEMICAL TESTING
Water samples may be examined using the principles of analytical chemistry.
pH, Biochemical oxygen demand Chemical oxygen demand nutrient estimation (nitrate phosphorus) Metals (copper, zinc, cadmium, lead mercury), hydrocarbons (oil and grease, petroleum ), pesticides.

Bacteriological water analysis is a method of analysing water to estimate the numbers of bacteria present
Multiple tube method ATP Testing Plate count Membrane filtration Pathogen analysis

Chemicals released into the environment forced indigenous microflora to -degrade compounds by producing
enzymes -detoxify by deriving metabolic energy

Petroleum hydrocarbons
-alkanes, aromatics, resins, asphaltenes -single carbon to 40 carbons -ranges from gas like methane to liquids to solids

n-alkanes are rapidly degraded

by - oxidation to form 2C shorter fatty acid and liberate CO2 and acetyl CoA

B-Oxidation of hydrocarbons

AROMATIC HYDROCARBONS Benzene ring hydroxylated ring cleavage two types - ortho cleavage - meta cleavage

Petroleum hydrocarbons are degraded by Bacteria

Pseudomonas spp. Micrococcus Achromobacter Nocardia Arthrobacter Acinetobacter Brevibacterium Corynebacterium Rhodotorula

Hydrocarbons are degraded by fungi

Aspergillus Penicillium Fusarium Gliocladium Trichoderma Candida Saccharomyces Rhodotorula Sporobolomyces

BIOREMEDIATION IN-SITU OR EX-SITU

Bioventing, Landfarming, Bioreactor, Composting, Bioaugmentation, Rhizofiltration, Biostimulation.

*Mycoremediation - use of fungal mycelia in bioremediation. Mycofilteration - use fungal mycelia to filter toxic waste

*95% of PAH (Polycyclic aromatic hydrocarbons) was reduced to nontoxic components in mycelia inoculated setups

A.M. CHAKRABARTY

Superbug OCT PLASMID alk BAC, alkR Degrades C6 to C10 n-alkanes

The sewage, river and floodwater discharged into the ocean carried plenty of terrestrial bacterial species along with it. It was thought that they rapidly disappeared when released in to sea (Carlucci and Pramer, 1959). Colwell and coworkers (1985) established that when enteric bacteria are introduced into sea water, they enter the viable non culturable state (VNC).
The VNC cells are viable with metabolic activity and were even found to have retained their virulence (Grimes and Colwell, 1986).

Van Overbeek and coworkers reported that P. fluorescens R2f cells could be detected with the immunofluorescence technique even in samples where no colonies were recovered through the conventional culture techniques.
The terrestrial bacteria when discharged in to sea water do not die immediately and act as donors of plasmids to indigenous marine bacteria.

The donor bacteria was isolated from soil samples collected from petroleum polluted sites around Madras Refineries. Potential hydrocarbon degrading culture of Pseudomonas fluorescens SKP3 was isolated by enrichment culture technique. The bacteria was resistant to wide range of antibiotics. The bacteria showed resistance to heavy metals also.

Table showing the antibiotic resistance/ sensitivity pattern of donor

Antibiotic Chloramphenica l Gentamycin Rifampicin Ampicillin Kanamycin Mitomycin C Tetracycline Streptomycin Benzylpenicillin Nalidixic acid Amoxycillin Erythromycin Penicillin G Cephalexin Vancomycin Cloxycillin MIC (g/ml) 575 70 15 375 65 5 40 325 275 550 375 575 525 725 775 LC (g/ml) <900 95 20 575 85 10 55 570 675 5 >900 685 >900 755 >900 >900

Plasmid DNA was isolated by the Casse et al. (1979) method. Conjugation was carried out by Broth mating method Plate mating method Filter mating method

The survival rate of non-halotolerant bacteria in marine water is not clearly known. In the present study, the survival rate of the donor is 3 to 4 weeks. Therefore, it would act as a potential donor of the plasmid to the marine bacteria.

The frequency of transfer is influenced by several environmental factors, like temperature and availability of nutrients. When the pool of resistance genes increases in the environment it would bring havoc to human health care.

- bioremediation. - plant growth stimulation. - biocontrol of pathogen

- either do well as hoped. - influence other harmful microorganisms - Transfer drug resistance to other microorganisms -overgrow indigenous strains. - bioterrorism.

3 Rs
Reduce Reuse Recycle

Kulithalai Municipality
*Solid Waste management *Segregation at source *Composting of degradable waste *Recycling of nondegradable waste