By Neha Mhapralkar Msc.

BT-12015

Mycoremediation is the process of degrading or removing toxicants from the environment using fungi. The term mycoremediation was coined by the American mycologist Paul Stamets, who has studied many potential uses of mushrooms for over 30 years, including their culinary, medicinal, and restorative properties. The process of mycoremediation is practised since the 20th century.

Fungi are the important decomposers in the natural environment. Fungi have proven to modify soil permeability and soil ion exchange to detoxify contaminated soil. They create enzymes to degrade plant polymers such as cellulose,hemicellulose and lignin. These enzymes break bond between hydrogen and carbon. As a result,mycoremediation may break down certain chemicals viz chlorinated pesticides which tend to persist in the environment. Bacteria can help to further degrade these compounds into final products including carbon dioxide (CO2), water, and potentially methane (CH4).

Fungi form four basic ecological roles; a)decomposers, b) mycorrhizal interaction, c) parasites, d) food source. Of these processes it is the ability of fungi to decompose that forms the integral part of mycoremediation, a process recognised as 'mycodegredation'. There is large characteristic variation between strains of fungi, with different strains able to breakdown and metabolise different compounds. They remove heavy metals from land by channeling them to fruit bodies for removal. they essentially use and digest these toxins as nutrients.

Mycelial enzymes can decompose some of the most resistant materials made by humans or nature, because many of the bonds that hold plant material together are similar to the bonds found in petroleum products including diesal, oil, and many herbicides and pesticides. The most developed branch of mycoremediation is concerned with 'white rot fungi'. The white rot fungi are a physiologically rather than taxonomically similar group of fungi which produce enzymes that break down lignin and other similar multi-chained compounds. Lignin is a complex chemical compound which is most commonly derived from wood, forming an integral part of the secondary cell walls of plants.

The soil food web showing the role of fungi as decomposers. Picture sourced from: http://www.sltec.com.au/

The white rot fungi are a physiologically rather than taxonomically similar group of fungi which produce enzymes that break down lignin and other similar multi-chained compounds. Lignin is a complex chemical compound which is most commonly derived from wood, forming an integral part of the secondary cell walls of plants. After the discovery of the extracellular oxidative ligninolytic enzymes of the white-rot fungus Phanerochaete chrysosporium, Bumpus et.al. (1985) proposed the use of this fungus for bioremediation. The white-rot fungus P.chrysosporium has emerged as an archetypal model system for fungal bioremediation. P. chrysosporium has the ability to degrade toxic or insoluble compounds more effi ciently than other fungi or microorganisms.

A variety of substrates, such as wood chips, wheat straw, peat, corncobs, sawdust are mixed with contaminated soils to enhance degradation. Three phases of strategies are envisioned for the successful implementation of mycoremediation. Inoculum preparation techniques and their improvements lead to success in the fi rst phase of the use of white-rot fungi in mycoremediation. The second phase includes clear technical protocols for the fi nal design and associated engineering processes. The remediation protocols for the monitoring, adjustment, continuity, and maintenance of the engineering system dictate the success of the third and fi nal phase of the mycoremediation process.

Mushrooms are voracious, and Tradd Cotter has found species that can chow down on E. coli and Salmonella, suck up lead, mercury, arsenic and other heavy metals, and break down chemicals into non-toxic compounds. Oyster mushrooms can break down 80% of DDT in 28 days and the aptly named "Train Wrecker" fungus can eat through railroad ties impregnated with CCA. The oyster mushroom Pleurotus ostreatus to metabolise multiple pollutant PAHs. Pleurotus ostreatus has a remarkable resistance to salty conditions, so much so that it is able to grow and reproduce in sea water. Lentinus edodes (shiitake mushroom) can degrade pentachlorophenol (PCP), a broad-spectrum biocide that is more toxic than DDT. Pleurotus pulmonarius (lung or Italian oyster mushroom) can degrade atrazine, a herbicide that is contaminating groundwater in many midwestern US states. There has been great interest by the public and government in using this technology to treat polluted environments in the wake of the Gulf of Mexico oil spill. The BP oil spill has inflicted enormous harm in the Gulf of Mexico and will continue to do so for months, if not decades, to come.

Mycoremediation has demonstrated positive results, verified by scientists in many countries. Oil contains a wide variety of toxins, many of which are carcinogens. Mycelium more readily degrades lower molecular weight hydrocarbons (3,4,5 ring) than heavier weight hydrocarbons. However, the heavier weight hydrocarbons are reduced via mycelial enzymes into lighter weight hydrocarbons, allowing for a staged reduction with subsequent mycelial treatments. Aged mycelium from oyster mushrooms (Pleurotus ostreatus) mixed in with compost' made from woodchips and yard waste resulted in far better degradation of hydrocarbons than oyster mushroom mycelium or compost alone. Worms die when put into contact with high concentrations of hydrocarbon saturated soils, but live after mycelial treatments reduce the toxins below the lethal thresholds.

A successfull technique has been to inoculate straw with fungi, then use the ability of straw to soak up and absorb oil from surrounding water.

Fig 1: Straw inoculated with fungal oil from water by straw inoculated with mycelium . mycelium.

Figure 2: Further absorption of by straw inoculat ed by

Fig 3: Oil being added to straw inoculated with mycelium

Figure 4: Oyster mushroom growing in the oil soaked straw

Fig 5: A mycoboom floating on salt water

Fig 6: Mushrooms growing from the end of a mycoboom

Fungal mycelium absorbing oil

There are numerous advantages of using mycoremediation over commercialized technologies, including the following: 1) Public acceptance:natural system,does not introduce any corrosives or other chemicals for cleanup. 2)Safety: safer than most other alternatives and it does not require digging up contaminated products, and disposing of it at waste sites. Additionally, the process does not produce secondary waste streams that require additional cleanup after the initial remediation. 3) Quiet: The technology is quieter than many alternatives, there are no structures, no machinery, and no noise. The system takes a day to set up, much like a landscaping project, and then left to do its work. 4) Low maintenance: There is minimal handling and low maintenance of sites treated with fungi.

5) Reusable end products: The end product of mycoremediation is nontoxic. The enriched and cleaned soil can be used for landscaping, road underlayment, or other purposes. 6) Low cost: The cost of using mycoremediation is relatively low in comparison to other technologies, as it it does not require building of new structures to house and process materials. 7) Flexible: The size of the application can vary without any problem, and can be the size of a bucket, to acres across. Additionally, fungal treatments can work in almost any habitat and season. 8) Fast: The technology shows immediate results. There is immediate mitigation of odor and visible improvement to a site. For end results, mycoremediation is quicker than other technologies, such as phytoremediation and bacterial bioremediation. These treatments may require one to three years or more, and cannot address all the contaminants that fungus can attack. Fungal treatment requires weeks to months.

Disadvantages are:1) Still in testing: Organizations that currently want to use the technology for cleanups are finding it a hard sell to their decision makers, as it is a technology that is unproven, and often times, those decision makers want to rely on proven technologies. 2) Applicability: There are many approaches to remediation; and certain ones are suitable in particular situations. 3) Efficiency level: Biological systems are never 100% efficient, which is difficult for some end-users to understand. 4) Surrounding environment: The use of a natural system can run into problems with the competitive natural environment in some areas, or with seasonal efficiency in extreme habitats.

Cost:The cost of using mycoremediation is expected to be lower than other technologies. There have been a few comparative studies that show that it will be much lower than that for any mechanical, chemical, electrical, thermal or other method that requires machinery, structures, facilities, power, or repeated application and maintenance. For starters, the materials needed to start the remediation (i.e. fungi) are less than other technologies. Additionally, treated soil in most studies has been proven to be cleaned up enough to reuse for landscaping, reducing costs for disposal and long term liability. So far, the mushrooms have shown no sign of contamination after doing their work. After treating the contamination, they decompose, preventing any costs related to disposal of the treatment material.

Mycoremediation has a wide range of applications. So far successful treatments have been conducted on; 1)industrial waste waters:Yeasts and fungi are used extensively to reduce the strength of a wide variety of foodprocessing wastewaters . The most commonly used yeast is Candida utilis, because of its ability to utilize a wide variety of carbon and nitrogen sources, its capacity to grow rapidly in high yields, and its tolerance of low pH. Fungi have the ability to convert biodegradable organic matter into a mycelium that not only has a high enough protein content to be valuable as an animal feed supplement.

2) DISTILLERY AND BREWERY WASTES: Distillery wastewater is produced as a result of distillation of ethanol produced in the fermentation of carbohydrates, which pose considerable problems in disposal or treatment. Various strains of the yeast Saccharomyces cerevisiae are most widely used because of their good growth rate and high ethanol tolerance S. cerevisiae has been shown to tolerate ethanol concentrations of upto 23% when grown under proper nutrient conditions. 3) METABOLISM OF PETROLEUM HYDROCARBONS: 4) DEGRADATION OF POLYCHLORINATED BIPHENYLS:

4) DEGRADATION OF POLYCHLORINATED BIPHENYLS: Fungal species belonging to several genera have the ability to grow on biphenyl and produce 2- and 4-hydroxybiphenyls and 4,4-dihydroxybiphenyl. 5) Pesticides.