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Submitted byin partial fulfillment of the award of the degree of Master of Technology in Environmental Engineering.

Sandip P. Ajmire ,

Technology that use plants to clean up contaminated sites.

remediation and restoration.

Green technology that uses plants systems for

Advantages Of Phytoremedition

Cost effective when compared to other more conventional methods. Nature method, more aesthetically pleasing, minimal land disturbance. It is more economically viable using the same tools and supplies as agriculture.

It is less disruptive to the environment and does not involve waiting for new plant communities to recolonise the site. Disposal sites are not needed. It is more likely to be accepted by the public as it is more aesthetically pleasing then traditional methods. It avoids excavation and transport of polluted media thus reducing the risk of spreading the contamination.

It has the potential to treat sites polluted with more than one type of pollutent

Disadvantages of Phytoremediation

It is dependant on the growing conditions required by the plant (i.e. climate, geology, altitude, temperature). Slow rate and difficult to achieve acceptable levels of decontamination. Potential phase transfer of contaminant. Possibility of contaminated plants entering the food chain. Possible spread of contaminant through falling leaves. Trees and plants require care. Success is dependant on the tolerance of the plant to the pollutant. Large scale operations require access to agricultural equipment and knowledge.

Contaminant solubility may be increased leading to greater environmental damage and the possibility of leaching.



1. Identification of plants & micro-aquatic plant for removal of municipal waste water. 2. Fabrication of experimental setup. 3. Fabrication of laboratory setup. 4. Conducting batch studies for the removal of pollutants from municipal waste water. 5. Conducting batch studies to find optimum opreting condition of various parameters.

CONVENTIAL METHOD Mechanical means are needed. For secondary or tertiary treatment of waste water is necessary. Needs very much cost to reach Standards for deposal.


Land for disposal is needed.

Mechanical means are not needed. For secondary or tertiary treatment of waste water is not necessary. 0.05 $/m3 needs to reach Standards for deposal. Land for disposal is not needed.

Multiple contaminants can be removed with the same plant.

Aquatic Macrophytes

Free-floating (very significant in tropics, particularly in

water bodies with fluctuating water levels) - Lemna (duckweed), Salvinia (water fern), Eichhornia (water hyacinth), Azolla (mosquito fern)

Rooted (confined to the shallow water) - Emergent: Typha (cattail), Phragmites (giant reed) - Submergent: Hydrilla, Utricularia (bladderwort)
- Floating-leaved: Nymphaea (water lily), Victoria amazonica (giant water lily)

Aquatic plants for wastewater treatment

Aquatic plants are chosen for absorb particular nutrient and to remove pathogens, metals and other contaminants from wastewater. Aquatic plants have been shown to be very effective as a secondary or tertiary state for water treatment and nutrient removal.

Scirpus lacustris , Typha latifolia , Egeria densa, Salvinia rotundifolia ,

Schoenoplectus lacustris,

Scirpus lacustris
Typha latifolia Egeria densa

Salvinia rotundifolia

Spirodela polyrhiza Canna flaccida Elodea nuttallii

Schoenoplectus lacustris

Pistia stratiotes Algae Phragmites australis Scirpus robustus

Myriphyllum aquaticum

Scirpus pungens

Typha capensis

Glyceria maxima

Lagorosiphon Major

Constructed wetlands or phytoremediation: This is a technology for treating wastewater. A constructed wetland consists of a gravel bed on which suitable wetland plants are grown. As water passes through the substrate, it is purified through the activity of bacteria attached to the gravel, plant roots, soil and other particles.

Lemna gives aesthetically pleasing, minimal land disturbance PHYTOREMEDATION

Lab Mechanisms

Continues Type Laboratory Experiment

How does it work?

- Plants in conjunction with bacteria and fungi in the rhizosphere transform, transport or store harmful chemicals. - Plants attributes make them good candidates root system surface area to absorb substances and efficient mechanisms to accumulate water, nutrients and minerals. selectively take up ions developed diversity and adaptivity to tolerate high levels of metals and other pollutants.


Phytotransformation/Phytodegradation pollutant is taken up by the plant and transformed in plant tissue (to be effective must be transformed to a less toxic form). Trichloroethylene (TCE), a prevalent ground water contaminant, transformed to less toxic metabolites by using hybrid poplar tree. Air Force facility in Texas using cottonwoods to treat a large ground water plume of TCE. EPA research lab using parrot feather (a common aquatic weed) for TNT treatment.


Uptake of chemical by the plant. Works well on metals such as lead, cadmium, copper, nickel etc. Detroit lead contaminated site was removed with Sunflower and Indian Mustard. - recently researchers at the University of Florida have determined that a species of fern, native to the south east, stores high concentrations of arsenic in its fronds and stems more than 200 times the concentration in the soil.


Vegetation holds contaminated soils in place - root system and low growing vegetation prevent mechanical transportation of pollutants from wind and erosion. - Trees transpire large quantities of water (more than 15 gal/day) so pumping action prevents contaminants from migration into the water table.

Use the extensive root system of plants as a filter. 1995, Sunflowers were used in a pond near Chernobyl - approx. 1 week they had hyperaccumulated several thousand times the concentration of cesium and strontium. - hyperaccumulation can contain 100 times or more of contaminant than normal plant.

Rhizosphere Bioremediation
- Increase soil organic carbon, bacteria, and mycorrhizal fungi, all factors that encourage degradation of organic chemical in soil. - The number of beneficial bacteria increased in the root zone of hybrid poplar trees and enhanced the degradation of BTEX, organic chemical, in soil.

Water Lily has an extensive root system with rapid growth rates, but is sensitive to cold temp, it is an ideal plant for water treatment in warm climates. Duckweed (Lemma spp.) has greater cold tolerance and a good capacity for nutrient absorption. Penny wort (Hydrocotyl spp) is relatively cold tolerant with a very good capacity for nutrient uptake. Water hyacint uptake of heavy metal eg.,Pb,Cu,Cd,Hg from contaminated water.

Aquatic plant for waste water treatment

Contaminant removal mechanisms

Sedimentation Filtration Adsorption Volatilization



Precipitation Bacterial metabolism Adsorption Plant metabolism Hydrolysis reaction Plant absorption Oxidation reaction Natural die-off


A suitable plant for rhizofiltration applications can remove toxic metals from solution over an extended period of time with its rapid-growth root system. Various plant species have been found to effectively remove toxic metals such as Cu2+, Cd2+, Cr6+, Ni2+, Pb2+, and Zn2+ from aqueous solutions. Low level radioactive contaminants also can be removed from liquid streams.

Rhizofiltration (cont.)

Limitations Rhizofiltration is particularly effective in applications where low concentrations and large volumes of water are involved. Data Requirements - Depth of contamination, - Types of heavy metal present, - Level of contamination must be determined and monitored. - Vegetation should be aquatic, emergent, or submergent plants. - Hydraulic detention time and sorption by the plant roots must be considered for a successful design.

Rhizofiltration (cont.)
The example of an experiment
The plant root immersed in flowing contaminated water until the root is saturated. The metal concentrated in the roots was analyzed on a dry weight basis using Atomic Absorption Spectrophotometry(AAS). The amount to metal taken up by the roots from various solutions was compared on the basis of recovery rate (g metal in roots/g metal in solution) and bioaccumulation coefficient (ppm metal in roots / ppm of metal in solution).

Rhizofiltration (cont.)

Other factors that should be considered

- Potential of failure modes and contigencies

Rhizofiltration may not succeed for a number of reasons, including mortality of plants for reasons such as management, weather extremes, soil conditions or pest. - Field studies Field studies are required before full-scale application. Specific information include rates of remediation, irrigation requirements, rates of soil amendments, and plant selection. Formulating clear objectives, appropriate treatments, experimental units and planning are important considerations in field studies. - Economic This technique should be less cost than traditional technologies such as excavation, thermal desorption, landfilling etc.


Although much remains to be studied, phytoremediation will clearly play some role in the stabilization and remediation of many contaminated sites. The main factor driving the implementation of phytoremediation projects are low costs with significant improvements in site aesthetics and the potential for ecosystem restoration.

Why Duckweed: (Lemna)

A small, floating aquatic plant that forms a solid cover on the surface of freshwater ponds, marshes, lakes and quiet streams Very important in the aquatic ecosystem as an essential link in the food chain.
Eaten by fish birds (ducks, herons)

Shades extensive areas of a pond, reducing algae growth Good for bioremediation projects

Ability to take out nitrogen and phosphorus from water

Useful as a water crop

as a source of food for animals and poultry

Structure of Duckweed

Leaves and stems merge in a common structure called a frond or thallus Has one root Fronds have 3 veins Air spaces help the plant to float


Reproduction is asexual Vegetative reproduction Unlike the leaves of more ordinary plants, each duckweed frond contains buds from which more fronds may grow. Until they mature, daughter fronds remain attach to the parent frond Rapidly growing plants can have 3-4 attached fronds

How do Duckweed Survive?

Make their own food through photosynthesis

Require sunlight Require nutrients

Phosphorous, nitrogen, other minerals

Need space to grow

Experiment: Population growth in Duckweed

1. 2. 3. 4.



Each group will set up TWO flasks Each flask has 100-200 ml pond water Place 2 duckweed plants in one flask Place 15 duckweed plants in the other flask Count all thalli (leaves) greater than 2 mm as full grown. Place labeled flasks under the grow lights in room 201 Record your data twice a week for 3-4 weeks (Create your own data table before you leave today)




Weight (g)







Commercial Feed

Commercial + Duckweed

Commercial + Periphyton

Duckweed + Periphyton

Introduction Aquaculture, the farming of aquatic organisms including fish and aquatic plants, is an alternative solution to reduce global reliance on wild fisheries. The rise in global population and subsequent increase in demand for consumable fish has put more stress on global fish stocks. The practice of farming fish is a logical alternative that would relieve much of the stress on global fisheries. Naylor, et. al. (2000) states that the long term growth of the aquaculture industry requires both ecologically sound practices and sustainable resource management. One of the most important problems facing aquaculture is the use of commercial feed, which is made up of fishmeal and fish oil. It is detrimental for the sustainability of aquaculture because it relies on wild fish stocks in order to feed farmed fish. The aquaponics system at Cape Eleuthera Institute (CEI) is the symbiotic cultivation of plants and aquatic animals in a re-circulating system. By combining aquaculture and hydroponics, nutrientrich wastewater from the fish tanks is circulated into the plant grow beds, which, in turn, sends filter oxygenated water back to the tanks. Currently the Tilapia in our system are being fed commercial feed containing fish meal and oil. The food is imported from Burris Specialty Feeds in Lousiana, which is a subsidiary of Cargill Incorporated. The purpose of our study is identify a locally grown, sustainable feed for the farming of Nile Tilapia (Oreochromis niloticus) in an aquaponics system. We hypothesize that if we feed O. niloticus with locally-grown plant-based feed sources, then their growth rates will meet or exceed current growth rates exhibited with commercial feed.

22.00 20.00 Average Weight (g) 18.00 16.00

a a b

14.00 12.00 10.00 1

Commercial Feed (Control)


4 Time (Weeks)


Methods The integrated aquaponics system combined the elements of aquaculture and hydroponics. The aquaponics system was composed of 12 buckets to test four treatments with two replicates. Each bucket had 10 Nile tilapia fingerlings (totaling 120 fish) with an initial weight of 15 0.36 g (mean standard deviation). The wastewater from the fish was filtered through a biofilter and lettuce bed, then into the duckweed trough. From there, the water was pumped back through the sump into the buckets. The daily parameters measured were temperature (C), dissolved oxygen (mg/L) and pH levels. Ammonia-nitrate levels were taken weekly. Each week all fish were removed to be weighed. These weights were used to calculate a mean biomass for each bucket and to calculate how much to feed those fish, based on feeding 5% of their body weight.

Duckweed was collected from a trough as part of the integrated aquaponics system and was subsequently dried, ground into a powder to emulate the texture of the commercial fingerling feed. Duckweed has a protein content of 33-38% (Tavares et. al 2008), in comparison to that of the commercial feed (Burris 35% protein).
The periphyton for this study naturally grows in waste water flowing into the mangrove adjacent to the cobia hatchery of Cape Eleuthera Institute. It was collected from rocks, rinsed, dried, ground into powder, and fed to the fish. Periphyton has a protein content of 12-30% (VanDam et. al 2002). The four treatments were commercial feed (the control), 50:50 duckweed:commercial, 50:50 periphyton:commercial, 50:50 periphyton:duckweed.


Through ANOVA testing, no statistical difference was found in mean weights between any treatment at the start of the study. Each treatment grew over the month-long period, but to varying degrees depending on the treatment (figure 1). The fish fed commercial feed (control) had the most growth, followed by the fish fed commercial:duckweed, then by the fish fed periphyton:duckweed. The fish fed commercial:periphyton exhibited the least growth, at times decreasing in biomass (figure 1). Fish fed commercial feed gained an average of 3.59 g. The resulting mean weight was 17.76 g (SD 0.91 g) (figure 4). It grew up until week three, but the mean weight declined from week three to week four by 1.07 g.
Fish fed commercial:duckweed feed had a mean growth of 2.98 g. The final mean weight was 17.68 g (SD 0.9 g). (figure 4). The p-value between this treatment and the control was p>0.05 so there was no statistical difference between the two treatments.
The fish fed commercial:periphyton feed had a mean growth of 1.20 g. The final mean weight was 16.22 g (SD 0.17 g) (figure 4). This treatment also had the most mortalities, ultimately resulting in its termination (figure 1 and 4). The p-value between this treatment and the control was p<0.05 g meaning the difference in growth was statistically significant.
The fish fed duckweed:periphyton had an increase in weight by 0.86 g. The final mean weight was 16.25 (SD 0.83 g) (figure 4). This treatment had a high standard deviation because each bucket had an outlier with a high weight. For this treatment and the control p=0.018, so the growth between each treatment was statistically significant. In conclusion, there was no statistical difference between the the control and commercial:duckweed meaning that the fish in both treatments exhibited similar growth perfomance. The fish fed commercial feed grew the most (figure 1) until week four when the final mean weight dropped. The fish fed commercial:periphyton stayed at about the same mean growths, as did the fish fed periphyton:duckweed which was supported by p=0.964.

Function of plants in aquatic treatment

Plant Parts

Uptake of pollutants surfaces on which bacteria grow media for filtration and adsorption of solids

and/or stem in water column

Stem and/or leaves at or above water surface

Attenuate sunlight, thus can prevent growth of suspended algae. Reduce effects of wind on water Reduce transfer of gases and heat between atmosphere and water.