Phytoremediation potential of Bermuda grass (Cynodon dactylon ) and Carabao grass ( Paspalum cojugatum ) in lead deposition : A comparative study ----------------------------------------------------------------------------An Undergraduate Thesis Presented to the Chemistry and Physics Department College of Arts and Sciences,Cebu Normal University -----------------------------------------------------------------------------In partial fulfillment of the Course Requirements For the degree Bachelor of Science in Chemistry and Physics
-------------------------------------------------------------------------------
Submitted by:
Anoc, Hannie Lou F. Rom,Sherlice Q.
March 2013
APPROVAL SHEET This research paper entitled “Phytoremediation potential of Bermuda grass (Cynodon dactylon) and Carabao grass (Paspalum conjugatum) in lead deposition: A comparative study” prepared by Hannie Lou F. Anoc and Sherlice Q. Rom. In partial fulfillment for the subject ChemPhys 114: Research in Chemistry and Physics has been examined and is recommended for acceptance and approval for Oral Examination. The Technical Panel JOYCE R. CALUMBA, MAST-Chemistry Chair, Chemistry and Physics Department GIBSON T. MAGLASANG, MS- Physics Professor, Chemistry and Physics Department KARL PATRICK R. CASAS, MS-Physics Professor, Chemistry and Physics Department DR. STELLA THERESE R. AVILA Chair, Biology Laboratory NIMFA PANSIT, MS Envi. Biology Professor, Biology Department ALLAN ROY ELNAR Professor, Chemistry and Physics Department Adviser Accepted and Approved in partial fulfillment of the requirements for the subject ChemPhys 114: Research in Chemistry and Physics.
FLORIZA N LAPLAP, Ed. D. Dean, College of Arts and Sciences Cebu Normal University i
PANEL OF EXAMINEES This research paper entitled “Phytoremediation potential of Bermuda grass (Cynodon dactylon) and Carabao grass (Paspalum conjugatum) in lead deposition: A comparative study” prepared by Hannie Lou F. Anoc and Sherlice Q. Rom. In partial fulfillment for the subject ChemPhys 114: Research in Chemistry and Physics passed on the Oral Examination and is approved by the committee of publishing.
JOYCE R. CALUMBA, MAST-Chemistry Chair, Chemistry and Physics Department
GIBSON T. MAGLASANG, MS- Physics Professor, Chemistry and Physics Department
KARL PATRICK R. CASAS, MS-Physics Professor, Chemistry and Physics Department
NIMFA PANSIT, MS Envi. Biology Professor, Biology Department
ALLAN ROY ELNAR Professor, Chemistry and Physics Department Adviser Accepted and Approved in partial fulfillment of the requirements for the subject ChemPhys 114: Research in Chemistry and Physics.
FLORIZA N. LAPLAP, Ed. D. Dean, College of Arts and Sciences Cebu Normal University ii
ACKNOWLEDGEMENT The accomplishment of this research won’t be possible without the unconditional support of our families, friends, panelists and those individuals that have shared their time with us while our research is on progress despite of their busy schedule. We would like to thank the following: To Mr. Allan Roy Elnar, our adviser, mentor, and a good father-like image to us, for his patience in checking and correcting our works to comply a much better and effective research paper; Mrs. Luzviminda Bato, for her heartfelt help to us, for being approachable to our concerns regarding our experiments; for keeping us motivated in our research and never stop believing in ourselves; Laboratory Custodians, Mr. Joenard Algones and Ms. Flor Marie Flores with their laboratory assistants, who willingly entrust to us their laboratories for us to use and for extending their laboratory duties in order for us to perform our experiment; Kuya Lester Jan R. Bato and Alden Deniega with the Technolab Analytic Group, for giving us lesser fees for the reading of our samples and analyzing them with precision; Kuya Tonyo, for offering his services and facilities open-hand while preparing and unloading some materials to be used in our experiment; Mr. Adonis Atuel, agriculturist, for sharing his experiences on planting grasses and advices that could help us in growing them; iii
Science Faculty, for sharing experiences in their respected field as a member of the Panel during our Thesis Proposal Hearing; Our beloved parents- Mama Alice and Papa Allan (Sherlice), and Mama Haydee and Papa Ronie (Hannie Lou), for their immeasurable support, emotionally and financially; from the start until the end of our study. Last but definitely not the least but the greatest, to God, for being with us in the ups and downs of our research, HE who has given us the perseverance on our research and the accomplishment of everything and finishing what we had started. Heartfelt grateful thanks to all of them who has been with us in our journey to our goal.
Hannie Lou and Sherlice
Cebu Normal University Cebu City March 2013
iv
ABSTRACT
The emergence of interests in phytoremediation studies were brought about by the increasing deposition of heavy metals and pollutants in the soil.In this study,Bermuda grass ( Cynodon dactylon ) and Carabao grass ( Paspalum conjugatum ) are compared based on their potential as lead phytoaccumulator at 3000 ppm and 6000 ppm concentrations.Grasses were grown for one month,prepared using acid-digestion and read by FlameAAS.The concentration of lead present in the amended samples shows that at 3000 ppm.It accumulate 5.6 mg/Kg of lead absorbed at a rejection percentage of 0.19%.These implied that carabao grass is a potential phytoremediating agent of lead while Bermuda grass does not exhibit the same potentiality.
Keywords: Bermuda grass,Carabao grass,Phytoremediation
v
Table of Contents Title Page Approval Sheet……….………………………………………………………………………………………….i Panel of Examinees………..…………………………………………………………………………………ii Acknowledgement…………..……………………………………………………………………………….iii Abstract…………………………….……………………………………………………………………………..iv Table of Contents…………….……………………………………………………………………………..vii List of Tables………………………………………………………………………………………………….viii List of Figures…………………..……………………………………………………………………………ix List of Appendices…………….…………………………………………………………………………….x ------------------------------------------------------------------------------------Chapter 1 Introduction…………………………………………………………………………………..1 Rationale……………………………………………………………………………………………….1 Statement of the Problem……………………………………………………………........3 Theoretical Background…………………………………………………………………………3 Scope and Limitations…………………………………………………………………………..9 Significance of the Study………………………………………………………………………9 Definition of Terms………………………………………………………………………………10 Chapter 2 Review of Related Literature…………………………………………………………12
vi
Chapter 3 Methodology…………………………………………………............................16 Research Design………………………………………………………………………...........16 Research Environment…………………………………………………………………………16 Research Procedure……………………………………………………………………………..16 Chapter 4 Results and Discussions……………………………………………………………….19 Results………………………………………………………………………………………………….19 Discussions…………………………………………………………………………………………..21 Chapter 5 Summary, Conclusion and Recommendations…………………………..25 Summary………………………………………………………………………………………………25 Conclusion…………………………………………………………………………………………….25 Recommendations………………………………………………………………………………..26 Bibliography……………………………………………………………………………………………………27 Appendices…………………………………………………………………………………………………….37 Curriculum Vitae…………………………………………………………………………………………….52
vii
LIST OF TABLES Tables 1.0
Page
Analysis of Variance One-Factor ANOVA result of Plant Growth……………………………………………………………………
2.0
Absorption of Lead through AAS (Atomic Absorption Spectrometer)…………………………………..
3.0
20
20
Analysis of Variance One-Factor ANOVA result of Lead Absorption……………………………………………………………….
viii
21
List of Figures Figures 1.0
Page Average Leaf Size (Carabao grass) and Diameter (Bermuda grass)……………………… 19
ix
List of Appendices Appendix
Page
A
Gannt Chart………………………………………………………………………………37
B
Plant Profile………………………………………………………………………………38
C
B.1
Bermuda grass……………………………………………………………….38
B.2
Carabao grass…………………………………………………………………39
Lead Concentration Accumulated using the Mean AAS (Atomic Absorption
Spectrometer)
reading………………….....40 D
Lead Accumulation and Rejection Percentage………………………..41
E
Preparation of 3000 and 6000 ppm Lead nitrate……………………42
F
Flame Atomic Absorption Spectrometer………………………………….43
G
Documentation…………………………………………………………………………45
H
Thesis Expenditures…………………………………………………………………48
I
Raw Data of Growth Rate ( Carabao and Bermuda Grass) ……………………………………………………………………49 J
Result of Analysis through AAS (Carabao grass)……………………………………………………………..50
K
Result of Analysis through AAS (Bermuda grass)…………………………………………………………...51
x CHAPTER 1 Introduction RATIONALE One of the many problems existing in the environment introduced in the soil is lead (Pb) (Ona et al., 2006 ).The main problem coexist with human activities such as mining (Liu et al.,2010),wet battery leaks (US EPA,2003 ),lead- waste water from industries (Ona et al., 2006 ),as well as heavy metal (e.g., lead (Pb), mercury (Hg),cadmium (Cd) ).While these problems continue to exist adverse health effects of heavy metal intake were caused
through
food
intake
from
plants
(
Nasreddine
and
Parent-
Massin,2002 ) and animals (Lead Poisoning in Livestock,2012). The accumulation of lead (Pb) in soil greatly affects the ecosystem. Since soils are considered as a major sink for lead it might be absorbed and bioaccumulated by plants and animals that may be available for human consumption in significant amounts (Effects of Lead in Plant Growth and Photosynthetic Activity, 2003).The general effect of lead (Pb) in plants is that it affects the physiological processes such as the slowing rate of photosynthetic activity and may lead to plant death. On the other hand, its effect on domestic animals took its effect in the central nervous system and inhibits the ability to synthesize blood cells according to US EPA (1986).And
on a report it generalizes that when animals are on a regular diet of 2-8 mg of lead per kilogram of body weight per day, over an extended period of time can cause death to most animals (US EPA, 1996). An alternative way of reducing lead (Pb) contamination is through phytoremediation. It is an alternative method that uses plants in cleaning up lead (Pb) contaminated areas (Uera et al., 2007). It is an easy to implement, cost-effective
and
an
environmentally-friendly
process
(Berti,
1997).
However, the success of phytoremediation depends on the choice of plant species, which can adapt and be relatively tolerant to the high concentration of heavy metals in soil (Uera et al., 2007).The study uses grass belonging to the Poaceae family. Grasses belongs to the plant family Poaceae in which according to recent studies is one of the 101 families which is known to be effective in metal hyperaccumulation (Kramer,2010).They are thought to be an excellent candidate in phytoremediation, because of their fibrous rooting system that can stabilize the soil and provide a large surface area for root-soil contact (Kulakow et al.,2000).Hence, the study is aimed to determine the potential of grasses as phytoremediating plants to lead (Pb).This further determines the level of concentration accumulated in the leaves. Consequently, it will
benefit the process of taking lead from contaminated soils, particularly in dumping sites. 2 STATEMENT OF THE PROBLEM The study aims to compare the lead accumulation of grasses namely: Carabao grass (Paspalum conjugatum) and Bermuda grass (Cynodon dactylon). Specifically it determines the following: 1. The level of lead (Pb) absorbance in grasses for the following treatment:
0 ppm of Pb (NO3)2
3000 ppm of Pb (NO3)2
6000 ppm of Pb (NO3)2
2. The effect of lead accumulation to growth rate of Carabao grass (Paspalum conjugatum) and Bermuda grass (Cynodon dactylon ).
Leaf size (Carabao grass)
Diameter (Bermuda grass)
3. The Rejection percentage (RejP) of the concentration of lead on the grasses being analyzed (see Theoretical Background for the formula). THEORETICAL BACKGROUND
The removal of heavy metals can be related to the following theory such as the Reverse Osmotic Theory. Here is the following explanation of the theory in the removal of heavy metals.
3 1. Phytoremediation Phytoremediation is the cleaning up of heavy metals in soil. Here are the mechanisms of phytoremediation. There are several types of phytoremediation processes that cover a large number of different organic and inorganic compounds. Only three are relevant to the phytoremediation of Lead (Pb).These three are termed (1) Phytoextraction – The uptake of contaminants by roots and translocation within the plants (2) Rhizofiltration – the adsorption or precipitation onto plant roots, or absorption into the roots of contaminants that are in solution surrounding the root zone, due to biotic or abiotic processes and (3) Phytostabilizationthe immobilization of a contaminant in soil through absorption and accumulation by roots, adsorption onto roots, or precipitation within the root zone of plants, and the use of plants and plant roots to prevent contaminant migration via wind and water leaching, and soil dispersion.
4 Source:http://www.scribd.com/doc/37203060/Phytoremediation-Technology-Hyper-Accumulation-Metals-in-Plants
Figure 1.0 Schematic representation of the accumulation of pollutants during phytoremediation (1)Phytoextraction. The uptake of contaminants by plant roots and translocation within the plants. It is primarily used in the treatment of soils,
Source:http://www.scribd.com/doc/37203060/Phytoremediation-Technology-Hyper-Accumulation-Metals-in-Plants
sediments and sludges. Constituents amenable to phytoextraction include : Metals – Ag, Cd, Co, Cr, Cu, Hg, Mn, Mo, Pb, Zn; Metalloids – As, Se; Radionuclides -90 Sr,137 Cs,239 Pu ,238 U,234 U. 5 Fodor’s model of the accumulation of heavy metals in plants The step-by-step process of the uptake of heavy metals on plants According to Fodor (2002) suggestion, the accumulation of heavy metal in plant is a stepwise process. Initially, is the interaction with other ionic components taking place at the locus entry into the plant rhizosphere that consequently have consequences for the metabolism. This is followed by an impact on the formation of reactive oxygen species (ROS) in the cell wall and an influence on the plasmalemma membrane system (stage 1).At stage 2,the metal ion reacts with all the possible interaction partners within the cytoplasm, including proteins, other macromolecules and metabolites. Stage 3 is mainly related to the factors that influence homeostatic events, which
include water uptake, transport and transpiration. At this stage, symptoms start to develop, and they become visible at stage 4.For instance, the chlorophyll and usually to a smaller degree, carotenoid content decrease, which have obvious consequences for photosynthesis and plant growth (Barcelo and Poschenrieder 2004).The death of plant cell occurs at stage 5.This model of Fodor has the advantage that visible effects are linked to metabolic events that are influenced by any metal ion. II. Reverse Osmosis Theory Reverse osmosis (RO) is a membrane process which initially was developed to produce potable water from saline and brackish water (Sirkar 6 et al., 1994).However, through the recent years of improvisation of its performance researchers begun to find out that it is not only applied in the treatment of water but as well as the recovery of organic and inorganic materials from chemical processes. Moreover, it can also remove organics, colour, nitrates and low total dissolved solids (TDS) concentrations (Sirkar et al., 1992).In like manner, removal of inorganic materials from soil requires grasses to have relatively high permeable membrane .Their efficiency can be described in the process of reverse osmosis (RO) (See Fig.3.0 ).
SOURCE: http://ph.images.search.yahoo.com/imags/view
Fig.3.0.Reverse Osmosis
Moreover, these physical characteristics implied the potential of plants in general as phytoremediating agents. Quantitatively, characteristics can be defined in terms of the plants rejection percentage (RejP) and recovery percentage (RecP). 7
Rejection Percentage is determined accordingly: CF -CP % RejP= ------------------ X 100 CF Where: CF – concentration of a specific component in the feed solution to the membrane process CP – concentration of the same specific component in the product stream leaving the membrane system On the other hand, recovery percentage (RecP) can be determined accordingly: CP
% RecP = -------- X 100 CF Where: CF – concentration of a specific component in the feed solution to the membrane process. CP – concentration of the same specific component in the product stream leaving the membrane system. In the case of concentration of lead, these percentages (RejP and RecP) can be translated to: Input concentration of lead in plants – output concentration of lead in plants
%RejP =------------------------------------------------------------------- X 100 Input concentration of lead in soil
8 %RecP is measured from the input and output concentration of lead as follows: Output concentration of lead in plants % Rec P = ---------------------------------------------- x 100 Input concentration of lead in soil SCOPE AND LIMITATIONS The study covers the comparative analysis among two kinds of grasses:
Carabao
grass
(Paspalum
conjugatum)
and
Bermuda
grass
(Cynodon dactylon ) and there efficiency in phytoremediation in the absorbance of Lead (II) nitrate Pb ( NO3)2.It also focuses on the effect of Lead (II) nitrate Pb ( NO3)2 to these grasses.
The delimination of the study is only two kinds of grasses will be analyzed: Carabao grass (Paspalum conjugatum) and Bermuda grass (Cynodon dactylon) and the contaminant used is Lead (II) nitrate Pb (NO 3)2 . The grasses to be used are young seedlings due to the unavailability of the seeds of grasses in the Philippines. The study limits on the comparison of the percentage of absorbance in Lead (II) nitrate Pb (NO 3)2 among the two grasses using Atomic Absorption Spectrometry (AAS). SIGNIFICANCE OF THE STUDY This study will beneficial to the following: (a)
PUBLIC – provide information about the phytoremediation potential
9 of selected grasses common in Cebu Provice such as Bermuda grass (Cynodon dactylon ) and Carabao grass (Paspalum conjugatum) accumulating Lead Nitrate (PbNO3)2. (b) COMPANIES – a tool in enhancing their waste management programs; provides an innovative, economical, and environmentallyfriendly alternative in removing toxic metals specifically Lead. (c) LOCAL GOVERNMENT – for the sustainable protection of the environment through strictly implementing the segregation of waste materials,
proper
disposal
through
having
garbage
cans
relocation sites for people living in dump sites for their safety.
and
(d) DENR – for the proper storage and disposal of the waste materials being handed to them for the safety of soil, plants and human health. (e) GRASS BREEDERS- for the awareness that these grasses: Carabao grass
(Paspalum
conjugatum)
and
Bermuda
grass
(Cynodon
dactylon) can help in the cleaning of the soil used in breeding plants. (f) DEPARTMENT OF AGRICULTURE- to breed more grasses especially for the benefit of the soil they will be using. DEFINITION OF TERMS
Grasses – belongs to the Poaceae family which is known to be effective in phytoremediation.
10 Reverse osmosis theory- a process in which it requires grasses to have relatively high permeable membrane to remove the inorganic materials found in the soil.
Potentiality of grasses – being adaptive and highly tolerant of grasses in the high concentrations of lead treated in the soil.
Phytoremediation – the use of plants in removing heavy metals in soil. The study focuses on the use of grasses as phytoremediating agent.
Heavy metals – hazardous in soil which is derived either naturally or chemically that maybe absorbed and bioaccumulated by plants and animals.
11 CHAPTER II Review of related literature and related studies The emergence of interests in phytoremediation studies were brought about by the increasing deposition of heavy metals and pollutants in the soil ( Caussy et al.,2003; Cui et al.,2004;Dudka et al.,1996;Muller and Anke, 1994;Sanchez – Camazano et al.,1994 ).In effect, ways to remove heavy metals
brought
hope
for
a
cleaner
environment
.However
,the
unprecedented urbanization and other anthropogenic human activities make
this likely impossible ( Dean et al.,1972;Dorsey,2003;Nriagu,1996;Mage et al.,1996;Pauleit et al.,2005;Ona et al., 2006;Randolph,2004;Widinarko et al.,2005
)
and
always
a
challenge
for
sustainable
development
(Cleverland,2003;Rees,1992).The many studies on removing heavy metals in the soil had been also a challenge because of the unbalanced rate of deposition and rate of removal (Singh et al.,2012).Also ,the methods used were costly (Cunningham et al.,1996;Singh et al.,2012 ). Moreover ,the deposition of lead (Pb) persisted for over 5000 years (Friedland,1990) and become the most common heavy metal contaminant in the soil ( Alloway,1995;EPA,1993; Wanatabe,1997 ).These contaminants were considered toxic to humans even when taken in minute amounts (Brinkmann,
1994;
Sheppard,
1998;
Thornton,
1991).In
addition,
leadcontamination prevailed due to existing mining and smelting activities
(Bridge,2004;Kodom
et
al.,2010;Lacatusu
et
al.,
2009;Nakayama
et
al.,2010;Nriagu,1996) as well as the use of paints ,gasoline, explosives, and the disposal of municipal sewage sludge and industrial wastes (FAO and WHO,2000;Reichman,2002;Zakrzewski,1991).These
activities
introduced
lead to the food chain and further into animals and human metabolism (ATSDR,2000;Sauve et al.,1997;Wang et al.,2001). The fatal effect of lead (Pb) intake into human metabolism includes seizures,
mental
retardation
(Canfield
et
al.,2003;Gosh
and
Singh,2005;Goyer,1993),behavioral disorders ( Gosh and Singh,2005) as well as brain and kidney damage ( Voroney,2006 ) and vomiting and appetite loss (FAO and WHO,2000;Mushak,1993).In like manner, lead (Pb) can lead to human genetic disorder, such as cancer (Beyersmann & Hartwig,2008;EPA Toxic Release Inventory ,2000 ).Consequently, the effects are irreversible ( Bellinger and Dietrich ,1994 ) that includes inhibited photosynthetic activities in plants and animals resulting from deficient mineral
intake
and
water
imbalance
(Adriano,1986;Afzal
et
al.,2006;Alloway,1990;Hao et al., 2004;Schmidt,2003;Sadiq,1992;Sharma et al.,2005;Wahla & Kirkham ,2008;WHO,1989). Lead
contaminated
soil
must
be
remediated
to
decrease
the
environmental risk. Many remediation techniques have been employed to address the rising number of heavy metal contaminated soils (Cholpecka et 13 al.,1996;Cunningham,1996;Cunningham
et
al.,1995
).
Most
of
the
traditional methods such as incineration, vitrification, electrokinetics and land filing are extremely expensive (Danh
et al., 2009;Mulligan et
al.,2001;Pulford and Watson,2003). Due to these problems the emergence of an environmentally friendly (Ranskin and Ensley, 2000) technology called Phytoremediation is widely accepted. Plants are attractive, economic and non-invasive alternatives to remove heavy metals (phytoextraction) from polluted soils as pointed out by
Blaylock and Huang (2000) and Salt et al. (1998).However, the plant species being used must grow well in toxic levels of heavy metal conditions and can produce high biomass (Berti, 2007).The success of phytoremediation is greatly dependent upon the choice of plant species to be used. The use of plants as agents to remove heavy metals includes spinach that can uptake a maximum of 192 µg g-1 Cadmium (Cd) at 50 µg g-1 treatment (Salaskar et al., 2011 ), radish according to Dean and Intawongse (2006)
can
accumulate
Copper
(Cu)
–
62.5%,Cadmium
(Cd)
–
54.9%,Manganese (Mn) – 45.8%, duckweed hyperaccumulates Cadmium (Cd),Copper (Cu) and Selenium (Se) (Lone et al.,2008 ) and as pointed out by Singh et al., (2012) it can remove up to 90% of soluble Lead (Pb) from water; and recently the work of Estrera and Banzon (2012) on yardlong beans used in Pb accumulation. 14 In similar manner the use of grasses as phytoremediating plants were studied by Sigua et al., (2007) and Xia (2003).They found out that the Vetiver grass had the potential in removing Pb ( Sigua et al.,2007;Xia ,2003) and Cadmium (Xia,2003).Accordingly, Vetiver grass are known for its effectiveness in erosion and sediment control ( Greenfield,1995 ),and highly tolerant to soil extreme condition ( Roongtanakiat and Chairoj,2001;Truong and Baker ,1996,1998;Truong,1999).These characteristics are an immediate requirement in removing heavy metals in soil. Therefore, the use of these
plants and perhaps variant grasses can be use as phytoremediation agent because of their relative tolerance to high concentration of heavy metals (Uera et al., 2007). Phytoremediation due to its low cost compared to the conventional cleaning-up technologies ( Chaney et al., 1997 ;Cunningham et al., 1996.1997;January,2006;Salt
et
environmentally
Chen
friendly
(
al.,1995;Sarma &
2011
)
and
Cutright,2002;Fayiaga
being et
al.,
2004;Pivertz,2001) is a very interesting topic for many researchers. Its great impact to our lives serves as a tool for a greener and healthier environment.
15 CHAPTER III METHODOLOGY RESEARCH DESIGN The study is experimental by nature. The amount of lead absorbed by the Bermuda grass and Carabao grass will be analyzed through AAS (Atomic Absorption Spectrometry). A 2X3 factorial experiment with two replications (Bermuda grass, Carabao grass ) per treatments ( 0 ppm,3000 ppm,6000
ppm ).The concentration of Lead accumulated by the two grasses is determined through the Rejection Percentage. RESEARCH ENVIRONMENT The study was performed at Cebu Normal University Chemistry Laboratory where the young grasses were being grown for one month. The grasses were then placed in aluminum foil pan and were arranged in blocks. After one month, the grasses were harvested and digested, and then the digested samples were forwarded at Technolab Analytical Group Inc. for the reading of the samples. RESEARCH PROCEDURE 1) Sample Germination and Collection The young grasses of Bermuda grass and Carabao grass that was grown for one month were labeled as (Control, T1 and T2), in which the control samples has 0 ppm lead amendment, while T1 and
T2 has the following amendment; 3000 ppm and 6000 ppm of lead, respectively. The grasses are allowed to grow for one month and the corresponding measurement of leaf size (Carabao grass) and diameter (Bermuda grass) were measured weekly. During the growth of the grasses, all treatment was watered with 2000 ml of distilled water (Ahmad et. Al., 2008) avoiding contamination aside from lead. The
grasses were then air-dried for one week after one month of germination. 2) Sample Preparation and Analysis a.) 0.20 g of each samples were placed in porcelain crucibles and were heated for 3 hours at 300 ˚C and an additional 2 hours at 500 ˚C inside a muffle furnace. Then 3 ml of 5 N Nitric acid was added to the samples and was heated at 200 ˚C for 15 minutes to remove traces of organic matter. Then, the samples were placed on the hot plate for drying followed by the addition of 5 ml 2 N Nitric acid to dissolve the residue of salts. The mixtures was filtered to catch its filtrate through a Whattman # 42 in a 250 ml volumetric flasks and then transferred to vials and was stored in the refrigerator ready for forwarding of analysis at Technolab Analytical Group Inc. b.) Analysis using Flame- AAS is based on the APHA AWWAWEF, Standard Methods for the Examination of Water and Wastewater, 17 21st Edition(American Public Association, 2005). 3) Statistical Analysis Statistical analysis employed in this study uses the software SPSS V16.
18 CHAPTER IV RESULTS AND DISCUSSION RESULTS I. GROWTH RATE
The Bermuda and Carabao grass was being observed in terms of its diameter and leaf size of the latter in which they were being measured weekly as shown in Figure 4.1. 300 250 200 CONTROL(in cm)
150
3000 ppm 6000 ppm
100 50 0 CARABAO(Leaf size)
TRIAL 2
BERMUDA(diameter)
TRIAL 2
Figure 1.0. Average Leaf size (Carabao) and Diameter (Bermuda) In reference to Figure 4.1, the control samples of carabao grass exhibit almost the same results in growth rate. The 3000 ppm amended sample show that the leaf size of the carabao grass grows increasingly. On the other hand, the 6000 ppm, however does not show the same result as with the first treatment, its leaf size grows increasingly but not as tall as the first treatment. On the other hand, Bermuda grass shows an almost the same diameter in both concentrations. TABLE 1.0. Analysis of Variance: One- Factor ANOVA result of plant growth Source of
SS
df
MS
F
P-value
F crit
Variation Between groups Within groups Total
3611.444
2
1805.72
0.34467
0.71391
3.6823
8
4
2
78583
1
2 5238.86
82194.44
5 1
7
4 7 F-value
%RejP
CARABAO GRASS Control 3000 6000 (mg/kg ppm ppm ) (mg/kg (mg/kg ) ) <0.03 5.6 <0.03 <0.03 0.19% <0.03
BERMUDA GRASS Control 3000 6000 (mg/kg ppm ppm ) (mg/kg (mg/kg ) ) <0.03 <0.03 <0.03 <0.03 <0.03
Table 2.0. Absorption of Lead through AAS(Atomic Absorption Spectrophotometer)
In the control samples of each grass it shows that it is <0.03 of lead concentration absorbed in which it the absorption value of the grass is less and nearly negligible. The concentration of lead present in the amended
20 samples of Carabao grass show that at 3000 ppm it accumulates 5.6 mg/kg of lead absorbed at a Rejection percentage of 0.19%(see Theoretical Background for the formula). In other concentration of lead, Carabao grass
and Bermuda grass shows the same results as to its controlled sample with <0.03 absorption. TABLE 3.0. Analysis of Variance: One- Factor ANOVA result of Lead Absorption Source
of SS
Variation Between
31.36
df
MS
F
P-value
F-crit
2
15.68
5
0.02168
3.68232
groups 4 Within groups 47.04 15 3.136 Total 78.4 17 F value>F-crit (5>3.68232) = Significant DISCUSSION I. GROWTH RATE The result as shown in Figure 1.0 represents the growth rate of each grass in a lead contaminant at different parameters in which there is a slight difference in their leaf size and diameter. According to Hasnain et al. (1995) and Prodgers and Inskeep (1981), with the increase of concentration and toxicity of heavy metals the growth of a plant gradually slows down its 21 growth rate which means it is concentration dependent (Miller and Koeppe, 1971). The slight difference of the plants’ growth is due to the limited test measuring in which only the selected parts of the plants are only measured (Banzon and Estrera, 2012).
The plant growth varies insignificantly, which implies that these grasses were tolerant at a certain concentration. Carabao grass easily adapts the presence of stress and developed resistance to lead without any harm to its growth and development implying that it is phytotolerant on an elevated Pb (Environmental Science Pollution Research, 2007). Though carabao grasses possess this kind of characteristic, but Cunningham and Ow(1996) exemplifies that heavy-metal stress can also be an attribution to the inhibition of plant’s growth and mechanisms. II. ABSORPTION AND ACCUMULATION OF LEAD The absorption of lead in grasses differs through its rooting system, since the Carabao grass has vigorous roots so it can absorb more Pb unlike Bermuda grass. Through this, it emphasizes the relationship between Pb accumulation and absorption and plant biomass in Pb extraction (Gray 2000). It has a direct relationship on its biomass and accumulation of metal;which means the more biomass the plant has the more metal it can accumulate, since the metal uptake is a function of the overall plant biomass claimed by Gray (2000) and as stated by the Environmental Science 22 Pollution Research (2007). Once lead has entered the root system it may accumulate there or translocated to other parts of the plants. For most plant species, the majority of absorbed lead is accumulated in the roots and only a fraction is
translocated to other parts of the plants as been reported to the studies of Piechalak et al.2002; Małecka et al.2008; Shahid et al.2011; Kopittke et al. 2007; Gichner et al.2008; Brunet et al. 2009; Gupta et al. 2009; Yan et al.2010; Gupta et al. 2010; Jiang and Liu 2010. The absorption of lead is less at 3000 ppm and other concentration is almost negligible, is due to the translocation of lead in the system in which the part of the grass being tested is where lead absorption is less. The uptake of nutrients and beneficial metals of plants is through its channels,
pores,
and
transporters
in
its
roots.
Through
it,
plants
characteristically have the capacity to absorb what they need and do not. However, most of vascular plants absorb toxic metals through their roots in varying degrees from negligible to substantial and sometimes there is absorption because of the chemical similarity between the beneficial and toxic chemicals. Some plants utilize exclusion mechanisms, in which there is a reduced uptake of roots or a restricted transport of the metal from roots to shoots (Baker 1981). The characteristic of being a phytoremediating agent is calculated 23 quantitatively through the rejection percentage (%RejP) and recovery percentage (%RecP), the study only limits on the former percentage. The percentage of 0.19% of lead in carabao grass at a concentration of 3000 ppm implies on how much lead is being absorbed by the grasses. In some
cases, in which Bermuda grass has 0 rejection percentage, this does not mean that no lead (Pb) is absorbed but of negligible amount. This implies that carabao grass is a potential phytoaccumulator of lead at 3000 ppm concentration while Bermuda grass does not possess the same characteristic with the latter, based on the result presented.
24 Chapter V SUMMARY, CONCLUSION AND RECOMMENDATIONS
SUMMARY
This study was conducted to compare the lead accumulation of Bermuda grass ( Cynodon dactylon ) and Carabao grass ( Paspalum conjugatum) in two different concentrations. After grown for one month, the plant where harvested; air dried and prepared using the Acid Digestion Method. The samples were tested for lead absorption usig the Flame Atomic Absorption Spectrophotometer (FAAS) and were statistically analyzed using One Factor ANOVA. Results showed that only Carabao grass at 3000 ppm accumulates lead with the amount of 5.6 mg/kg. This result implies that the carabao grass is an accumulator of lead, due to its vigorous rooting compared to Bermuda grass. CONCLUSION The amount of lead accumulated may have been very negligible due to the
rooting
system
of
each
grass.
Carabao
grass
is
a
potential
phytoremediating agent of lead at a concentration of 3000 ppm while Bermuda grass does not exhibit the same potentiality.
RECOMMENDATIONS The researcher’s scope of study limits the duration of experiment weight of sample used and the equipment used for the analysis and hence, would suggest the following improvements of this study.
One of the first considerations in this study is its environment; there must be a consistency in the location to avoid other aspects that can affect the study. Second, is to prolong the duration of experiment for about 3 months. This is to ensure that there is a clear difference between the amount concentrations absorbed by these grasses. Second, is to increase the dry weight of the samples used for analysis since the minimum amount needed for accurate reading in Flame-AAS is 0.5 g. Making the reading of accumulated lead increase and not on a negligible amount. Third, is the equipment used for more accurate reading, for a better reading of the samples
we
suggested
to
use
a
Graphite
Atomic
Absorption
Spectrophotometer that can determine metals in quantities as low as 10 -12 g. Fourth, is the proper disposal of lead with the help of Department of Environment and Natural Resources (DENR).
26 BIBLIOGRAPHY Adriano, D. C. Trace Elements in the Terrestrial Environment. New York : Springer-Verlag. [Online]1986. Agency for Toxic Substances and Disease Registry (ATSDR).
Toxicological profile for Polychlorinated Biphenyls (PCBs). Atlanta, GA: U.S. Department of Health and Human Services, Public Health Service. [Online] 2000. Ahmad, R., M.; Arshad, Z.A.; Zahir, M. Naveed; M. Khlaid and H.N. Asghar. Integrating N-enriched compost with biologically active substances for improving growth and yield of cereals. Pak. J. Bot. [Online] 2008. 40(1): 283-293. Afzal, I., S.M. A. Basra, A. Hameed and M. Farooq. Physiological enhancements for alleviation of salt stress in wheat. Pak. J. Bot.[Online]2006,38: 1649-1659. Alloway, B.J. Heavy metals in soils. John Wiley and Sons, Inc. New York, ISBN 0470215984.[Online] 1990. Alloway, J. B.Soil Pollution and Land Contamination, in Pollution: Causes, Effects and Control, ed. R. M. Harrison. [Online] 1995. Baker AJM .Accumulation and excluders-strategies in the response of plants to heavy metals. Journal of Plant Nutrition [Online] 1981,3: 643–654. Barcelo J;Poschenrieder C. Structural and ultrastructural changes in heavy metal exposed plants. In: Prasad MNV (ed) Heavy metal stress in plants, 3rd edn. Springer, Berlin [Online] 2004, pp 223–248. Bellinger D, Dietrich KN. Low-level lead exposure and cognitive function in children. Pediatr Ann [Online] 1994;23:600–605. Beyersmann D.;Hartwig A.Carcinogenic metal compounds : recent insight into molecular and cellular mechanisms.Arch toxicol [Online] 2008,82:493-512. Blaylock, M.J, and J.W. Huang,.Phytoextraction of metals, In: I. Raskin and B.D. Ensley (Ed.) Phytoremediation of toxic metals: using plants to clean up the environment. John Wiley and SonsInc, Toronto, Canada [Online] 2000, p. 303. Bridge G . Contested terrain: mining and the environment. Annu. Rev. Environ. Resour. [Online] 2004,29: 205-259. Brinkmann, R. Lead pollution in soils adjacent to homes in Tampa,
Florida. Environmental Geochemistry and Health [Online]1994 , 16 (2): 59-64. BrunetJ,VarraultG,ZuilyFodilY,RepellinA.Accumulation oflead in the rootsofgrasspea(LathyrussativusL.)plants triggers system icvariation in gene expression in the shoots.Chemosphere 2009,77(8):1113-1120. Canfield RL;Henderson CR; Cory-Slechta DA;Cox C;Jusko TA; Lanphear BP. Intellectual impairment in children with blood lead concentrations below 10 micrograms per deciliter. N Engl J Med. [Online]2003,348:1517–1526. Caussy D.;Gochfeld M.; Gurzau E.; Neagu C. and Ruedel H. Lessons from case studies of metals: investigating exposure, bioavailability, and risk.Ecotoxiclogy and Environmental Safety [Online] 2003, 56: 45-51. Chaney, R.L; M. Malik; Y.M. Li;S.L. Brown, J.;S.Angle and A.J.M. Baker..Phytoremediation of soil metals. Current Opinions in Biotechnology [Online] 1997, 8 : 279-284. Chen, H., and Cutright, T.The interactive effects of chelator, fertilizer, and rhizobacteria for enhancing phytoremediation of heavy metal contaminated soil. J. Soil. Sediment [Online] 2002,2:203210. Cholpecka A; Bacon JR; Wilson MJ;Kay J .Heavy metals in the environment. Forms of cadmium, lead, and zinc in contaminated soils from southwest Poland. J Environ Qual 1996,25: 69– 79. Cleveland, C.J.Biophysical constraints to economic growth, In: D. Al Gobaisi, Editor-in-Chief, Encyclopedia of Life Support Systems. EOLSS Publishers Co., Oxford, UK. [Online] 2003. 28 Cui,Y.J.;Zhu,Y.G.;Zhai,R.H.;Chen,D.Y.;,Huang,Y.Z.;Qiu,Y.;Ling,J.Z. Transfer of metals from soil to vegetables in an area near a smelter in Nanning,China. Environment International [Online] 2004,30,785e791. Cunningham SD .Promise and prospects for phytoremediation. Plant Physiology [Online] 1996, 110: 715–719
Cunningham, S. D.;T. A. Anderson, A.; P. Schwab, and F. C. Hsu.Phytoremediation of oils contaminated with organic pollutants. Advances in Agronomy [Online] 1996,56: 55-107. Cunningham SD; Berti WR;Huang JW. Phytoremediation of contaminated soils. Trends Biotechnol [Online] 1995,13: 393–397. Cunningham, S.D.; Shann, J.R.; Crowley, D.E. and Anderson, T.A. Phytoremediation of contaminated water and soil. In: KRUGER, E.L.; ANDERSON, T.A. and COATS, J.R. eds. Phytoremediation of soil and water contaminants. ACS symposium series 664. Washington, DC, American Chemical Society [Online]1997,2-19. Danh, L.T.; P. Truong, R. Mammucari; T. Tran and N. Foster, 2009. Vetiver grass, Vetiveria zizanioides: A choice plant for phytoremediation of heavy metals and organic wastes. Int. J. Phytorem [Online] 2009, 11: 664-691. Dean, J.G.; F.L. Bosqui, and K.H. Lanouette. "Removing Heavy Metals from Wastewater".Environ. Sci. Technol [Onlline ] 1972, 6(6): 518-522. Dean, J. R ;Intawongse, M. Uptake of Heavy Metals by Vegetable Plants Grown on Contaminated Soil and their Bioavailability in the Human Gastrointestinal Tract. Food Additives and Contaminants [Online]2006, 23,pp 36-48. Department of Primary Industries,Victoria,Australia.Lead Poisoning in Livestock;htpp:/www.dpi.vic.gov.au/agriculture/pests-diseasesand-weeds/animal-diseases2/lead-poisoning-in-livestock (accessed August 2012 ). 29 Divya Singh ;Archana Tiwari and Richa Gupta . Phytoremediation of lead from wastewater using aquatic plants. Journal of Agricultural Technology [Online ] 2012, 8(1): 1-11. 29 Dorsey, J. Brownfields and Greenfields: The Intersection of Sustainable Development and Environmental Stewardship.Environmental Practice [Online] 2003,51: 69-76. Dudka S.;Piotrowska M.; Terelak H. Transfer of cadmium, lead and zinc from industrially contaminated soil to crops plants: a field study. Environmental Pollution [Online] 1996, 94:
181–188. Environmental Science Pollution Research.[Online] 2007. EPA. Clean Water Act, sec. 503, vol. 58, no. 32. (U.S. Environmental Protection Agency Washington, D.C.). [Online] 1993. Estrera J.;Banzon J.Phytotolerance of Yard Long bean (Vigna inguiculata subsp.sesquipedalis ) to lead.Cebu Normal University 2012. Fayiga,A.O;Ma,L.Q.;Cao,X.;Rathinasabapathi,B..Effects of Heavy Metals on growth and arsenic accumulation in the arsenic hyperaccumulator Pteris vittata L.Environ.Pollut [Online]2004,132:289–296. Friedland A.J. The movement of metals through soils and ecosystems. In : Shaw AJ (ed) Heavy metals tolerance implants : evolutionary aspects. CRC Press, Boca Raton[Online]1990, 7-19. Fodor F. Physiological responses of vascular plants to heavy metals. In: Prasad MNV, Strzalka K (eds) Physiology and biochemistry of metal toxicity and tolerance in plants. Kluwer Academic Publisher, Dortrech [Online] 2002, pp 149–177. GichnerT;ZnidarI;SzákováJ.Evaluation of DNA damage and mutagenicityinducedbyleadintobaccoplants.Mutat Res Genet Toxicol EnvironMutagen6522008,(2):186-190. Ghosh, M. and S.P. Singh.A review on phytoremediation of heavy metals and utilization of its byproducts. Applied Ecology Environmental Research [Online]2005, 3: 1-18. Goyer, R.A. Lead toxicity: current concerns. Env. Health Perspect. [Online]1993,100:177-187.
30 Gray KA.The phytoremediation of lead in urban,residential soils.Northwestern University,Evanston,Illinois [Online] 2000. Greenfield JC: Vetivergrass, the ideal plant for vegetative soil and moisture conservation. The World Bank, Washington, DC [Online] 1995,pp 31–48.
and
GuptaD;NicolosoF;SchetingerM;RossatoL;PereiraL;CastroG;Srivastava S;TripathiR. Antioxidant defense mechanism in hydroponically grown Zeamays seedlings under moderate lead stress.JHazardMater [Online]2009,172(1):479-484. GuptaD,HuangH,YangX,RazafndrabeB,InouheM.The detoxification of lead inSedumal fredii H.is not related to phytochelatins but the glutathione.JHazardMater177 [Online] 2010,(1(3)):437-444. Hao, X.Z., D.M. Zhou and Y.B. Si. Revegetation of copper mine tailings with ryegrass and willow. Pedosphere [Online]2004,14: 283288. Hasnain,SH;Sari,N.;Salaeem,F.Biotechnology for environment and agriculture.University of Karachi ,Pakistan. Inskeep,WP;Prodgers,RA.Heavy metals tolerance of inland saly grass Distilchlis spicata.Great Basin Naturalist [Online] 1981,51:271278. January MC .Hydroponic Phytoremediation of Cd (III), Cr (III), Ni (II), As (V), and Fe (II) by HELIANTHUS ANNUUS. M.Sc. Thesis, The Graduate Faculty of The University of Akron [Online]2006. JiangW;LiuD.Pb induced cellular defense systemin the root meristematic cells of Allium sativum L.BMCPlantBiol 2010,10:40-40.
[Online]
Kodom K, Wiafe-Akenten J, Boamah D . Soil Heavy Metal Pollution along Subin River in Kumasi, Ghana; Using X-Ray Fluorescence (XRF) Analysis. Conference Information: 20th International Congress on X-Ray Optics and Microanalysis, X-ray Optics and Microanalysis, Proceedings Book Series: AIP Conf. Proc. [Online] 2010,1221: 101-108. 31 Kopittke, PM; Asher, CJ; Blamey, FPC; Menzies, NW. Toxic effects of Pb2+ on the growth and mineral nutrition of signal grass (Brachiaria decumbens) and Rhodes grass (Chloris gayana). PLANT AND SOIL. 2007,300(1-2):127-136. Kramer, U. Metal hyperaccumulation in plants. Annu. Rev. Plant Biol. [Online] 2010, 61:7-534.
Kulakow, P.A;A.P. Schwab and M.K. Banks. Screening plant species for growth on weathered, petroleum hydrocarbon-contaminated sediments. Int. J. Phytorem.[Online] 2000, 2: 297-317. Lacatusu R, Citu G, Aston J, Lungu M, Lacatusu AR .Heavy metals soil pollution state in relation to potential future mining activities in the Rosia Montana area. Carpathian J. Earth Environ. Sci.2009, 4: 39-50. Liu, S.; Xia, X.; Yang, L.; Shen, M. & Liu, R. Polycyclic aromatic hydrocarbons in urban soils of different land uses in Beijing, China: Distribution, sources and their correlation with the city’s urbanization history. Journal of Hazardous Materials [Online]2010, 177: 1085–1092. Lone MI; He Z;Stoffella PJ;Yang X .Phytoremediation of heavy metal polluted soils and water: Progresses and perspectives, J. Zhejiang Uni. Sci. B [Online] 2008, 9: 210-220. MaleckaA;PiechalakA;MorkunasI;TomaszewskaB.Accumulation of lead in root cells of Pisumsativum.ActaPhysiolPlant [Online]2008,30(5):629-637. Miller RJ and Koeppe DE. Southern corn leaf blight: susceptible and resistant mitochondria. Science.[Online] July 1971,173(3991):67–69. Muller, M. and M. Anke.Distribution of cadmium in the food chain (soilplant-human) of a cadmium exposed area and the health risk of the general population. Sci. Total Environ.[Online] 1994,25:151158. Mulligan CN; Yong RN, Gibbs BF.Remediation technologies for metalcontaminated soils and groundwater: an evaluation. Engineering Geology. 2001, Vol.60, pp. 19-207. 32 Mushak P.New directions in the toxicokinetics of humanlead exposure.Neurotoxicology [Online]1993;14:29–42. Nakayama MMS, Ikenaka Y, Muzandu K, Choongo K, Oroszlany B, Teraoka H, Mizuno N, Ishizuka M. Heavy Metal Accumulation in Lake Sediments, Fish (Oreochromis niloticus and Serranochromis thumbergi), and Crayfish (Cherax
quadricarinatus) in Lake Itezhi-tezhi and Lake Kariba, Zambia. Arch. Environ. Contaminat. Toxicol [Online]2010,59(2):291-300. Nasreddine L; Parent-Massin D. Food contamination by metals and pesticides in the European Union. Should we worry? Toxicol Lett [Online]2002,127(1-3):29-41. Nriagu JO .Toxic metal Pollution in Africa. Science [Online] 1996,223: 272. Nriagu,Jerome.A History of Global Metal pollution.Science New Series 1996 [Online] January 2005,272:223-224. Ona LF; Alberto AP;Prudente JA; Sigua GC: Levels of lead in urban soils from selected cities in the rice-based region of the Philippines. Env Sci Pollut Res [Online] 2006,13 (3) 177–183. Pauleit, S.; Ennos, R.;and Golding, Y. Modelling the environmental impacts of urban land use and land cover change: a study in Merseyside UK. Landscape and Urban Planning [Online] 2005, 71, 2-4, 295-310. PiechalakA;TomaszewskaB;BaralkiewiczD;MaleckaA.Accumulation and detoxification oflead ions in legumes.Phytochemistry [Online] 2002,60(2):153-162. Pivetz, RE.Phytoremediation of Contaminated Soil and Ground Water at Hazardous Waste Sites. EP A/540/S-0 1/500.[Online] 2001. Pulford, I.D.; Watson, C.Phytoremediation of heavy metalcontaminated land by trees :a review. Environ. Int.[Online] 2003, 29: 529-540. Randolph J.Environmental land use and planning management. Island Press suit [ Online ] 2004. 33 Raskin I ;Ensley BD.Phytoremediation of toxic metals using plants To clean up the environment.NewYork:Wiley[Online]2000,316. Rees, W. Ecological footprints and appropriated carrying capacity: What urban economics leaves out. Environment and Urbanization [Online] 1992,4(2):121-130.
Reichman, S.M.The Responses of Plants to Metal Toxicity.A Review Focusing on Copper,Manganese and Zinc .AMEEF Paper 14. 59 pp. Australian Minerals and Energy Environment Foundation, Melbourne [Online] 2002. Roongtanakiat, N.and Chairoj, P.Uptake potential of some heavy metals by vetiver grass. Kasetsart J. (Nat. Sci.) 2001,35 : 4650. Sadiq,M. Toxic Chemistryin Marine Environments.Marcel DekkerInc.,NewYork.[Online] 1992. Salaskar, Darshana; Shrivastava, Manoj; Kale, Sharad P. Bioremediation potential of spinach (Spinacia oleracea L.) for decontamination of cadmium in soil. Current Science (00113891) [Online] November 2011, 101: 10 p1359. Salt, D.E.; Smith, R.D. and Raskin, I. Phytoremediation. Annual Review of Plant Physiology and Plant Molecular Biology[Online] 1998, 49: 643-668. Salt, D.E.;M. Blaylock;P.B.A.N. Kumar; V. Dushenkov; B.D. Ensley, I. Chet, and I. Raskin: Phytoremediation: A novel strategy for the removal of toxic metals from the environment using plants. Biotechnol [Online] 1995,13, 468-475 . Sanchez-Camazano M; Sanchez-Martin MJ; Lorenzo LF .Lead and Cadmium in soils and vegetables from urban gardens of Salamanca (Spain).Science of the total environment [Online]1994,pp 163-168. Sarma H. Metal Hyperaccumulation in Plants: A Review Focusing on Phytoremediation Technology. Journal of Environmental Science and Technology [Online ] 2011, 4: 118-138. 34 Sauve S;McBride M B;Norvell W and Hendershot W H Copper solubility and speciation of in situ contaminated soils: Effects of copper level, pH and organic matter. Water Air Soil Pollut [Online]1997 ,100:132–149. Schmidt U. Enhancing Phytoextraction, The effect of chemical soil manipulation on mobility, plant accumulation, and leaching of
heavy metals. J Environ Qual [Online] 2003, 32:1939-1954. ShahidM,PinelliE,PourrutB,SilvestreJ,DumatC.Lead induced genotoxicityto Viciafaba L.roots in relation with metal cell uptake and initial speciation.EcotoxicolEnvironSaf [Online] 2011,74(1):78-84. Sharma J., Saini V., Singh A., Singh N., Trivedy R.K, Sharma S. (Eds) Phytoremediation of organic pollutants , Biotechnological applications in environmental management.B.S.publication [Online] 2005. Sheppard, S.C. Geophagy:Who Eats Soils and Where Do Possible Contaminants Go?.Environmental Geology (Berlin).[Online] 1998 ,33(2-3):109-114. Sigua, Gilbert; Paz-Alberto, Annie; Uera, Raynato. Phytoremediation potentials of selected tropical plants for Ethidium bromide. Environmental Science & Pollution Research [Online] 2007, 14 : 7 505. Thornton, E.; Daview, D. J. A.; Watt, J. M.; Quinn, M. J. Lead exposure in young children from dust and soil in the United Kingdom. Environ. Health Persp.[Online] 1991, 89: 55-60. Troung PN. Application of the vetiver system for phytoremediation of mercury pollution in the Lake and Yolo countries. Northern California. [Online]1999. Truong, P. and Baker, D. Vetiver grass for stabilization of acid sulfate soil, In Proc. 2 Nat. Conf. Acid Sulfate Soils. Coffs Harbour, Australia. [Online] 1998,196-198. Troung PN and Baker D .Vetivergrass system for environmental protection. Royal Development Projects Protection.Technical Bulletin No. 1998/1. Pacific Rim Vetiver. Office ofthe Royal Development Projects Board, Bangkok, Thailand [Online]1996. 35 Uera RB;Paz Alberto AM;Sigua GC. Phytoremediation potentials of selected tropical plants for ethidium bromide.Environ Sci Pollut Res Int. [Online] November 2007, 14(7):505-9. US-EPA.National Emission Standards for Hazardous Air Pollutants for Coke Ovens: Pushing, Quenching, Battery Stack (66FR35326). US Environmental Protection Agency [Online] 2003.
Wahla IH;Kirkham MB. Heavy metal displacement in salt-waterirrigated soil during phytoremediation.Environ pollut.[Online] 2008 , 155(2):271-83. Wang H, Kimberley MO, Schlegelmilch M.Biosolids derived nitrogen mineralization and transformation in forest soils. J. Environ. Qual. [Online]2001,32:1851–1856. Watanabe, M.E. Phytoremediation on the brink of commercialization. Environmental Science & Technology/News. [Online] 1997,31:182-186. WHO. LEAD Action News. The journal of The LEAD (Lead Education and Abatement Design) Group Inc. [Online] 1989, 1 no 2 Winter ISSN 1324-6011. Widinarko et al. Spatial Relationships of Urban Land Use, Soils and Heavy Metal Concentrations in Lagos Mainland Area. J. Appl. Sci. Environ. Manage.[Online]2005. Xia H. Vetiver system for land reclamation. The third international conference on Vetiver, Guangzhou, China [Online]2003. YanZZ;KeL,TamNFY.Lead stress in seedlings of Avicennia marina,a common mangrove Species in South China,with and without cotyledons.AquatBot [Online]2010,92(2):112-118. Zakrzewski, S.F. Principles of environmental toxicology. ACS Professional Reference Book. Washington, DC.: American Chemical Society. [Online] 1991.
36 APPENDIX A GANNT CHART MONTH 2012
2013
Topic searching Title/proposal making Write-ups ( Chapter 1-III ) Thesis proposal hearing Preparation for experiment Planting and height measurement Harvesting and drying Sample preparation/AAS Analysis Analysis of Results Thesis writing Proof reading and Final Defense
37 APPENDIX B APPENDIX B.1 PLANT PROFILE
MARCH
FEBRUARY
JANUARY
DECEMBER
NOVEMBER
OCTOBER
SEPTEMBER
AUGUST
JULY
JUNE
ACTIVITY
Common Name: Bermuda grass Scientific Name: Cynodon dactylon Classification: Kingdom
Plantae – Plants
Subkingdom Superdivision Division Class Subclass Order Family Genus Species
Tracheobionta – Vascular plants Spermatophyta – Seed plants Magnoliophyta – Flowering plants Liliopsida
– Monocotyledons
Commelinidae Cyperales Poaceae – Grass family Cynodon Rich. Cynodon dactylon 38 APPENDIX B.1 PLANT PROFILE
Common Name: Carabao grass Scientific Name: Paspalum conjugatum Classification: Kingdom
Plantae – Plants
Subkingdom Superdivision Division Class
Tracheobionta – Vascular plants Spermatophyta – Seed plants Magnoliophyta – Flowering plants Liliopsida – Monocotyledons
Subclass Order Family Genus Species
Commelinidae Cyperales Poaceae – Grass family Paspalum L. – crowngrass Paspalum conjugatum 39 APPENDIX C
Lead Concentration Accumulated using the Mean of AAS Reading
Formula :
Lead concentration = AAS reading x Volume of Prepared Sample used kg dry weight dry weight used
Control
Lead concentration = Kg dry weight
0 μg x 50 ml mL________ = 0 μg/g = 0 mg/kg 0.20 g
Amended
Lead concentration = Kg dry weight
0.024 μg x 50 ml mL________ __= 5.6 μg/g = 5.6 mg/kg 0.20 g
40 APPENDIX D Lead Accumulation & Rejection Percentage SAMPLES Bermuda Grass
Total lead Concentration
Accumulated Concentration
Rejection Percentage
Present in soil (ppm )
in ppm
(%)
CONTROL ( 0 ppm )
<0.03
<0.03
<0.03
AMENDED ( 3000 ppm )
3000
<0.03
<0.03
AMENDED ( 6000 ppm )
6000
<0.03
<0.03
CONTROL ( 0 ppm )
<0.03
<0.03
<0.03
AMENDED ( 3000 ppm )
3000
5.6
0.19
AMENDED ( 6000 ppm )
6000
<0.03
<0.03
Carabao Grass
Input concentration of lead in soil – output concentration of lead in plants
%RejP =------------------------------------------------------------------- X 100 Input concentration of lead in soil 41 APPENDIX E Preparation of 3000 ppm and 6000 ppm Lead nitrate
“Parts per million” – usually abbreviated as “ppm” – means “out of a million” (ppm) commonly used in measuring small levels of the amount of pollutants in air, water and body fluids etc. 1 ppm= 1 mg/L =1 mg/kg Parts per million is the mass ratio
between
the
pollutant
component and the solution and ppm is defined as ppm = 1,000,000 mc / ms where mc = mass of component (kg, lbm) ms = mass of solution (kg, lbm) In the metric system ppm can be expressed in terms of milligram versus kg where 1 mg/kg = 1 part per million
42 APPENDIX F Flame Atomic Absorption Spectrometry (FAAS)
http://www.etslabs.com/images/methods/6.gif Atomic Absorption Spectroscopy is a technique for determining the concentration of a particular metal element within a sample. Atomic absorption spectroscopy can be used to analyze the concentration of over 62 different metals in a solution. (See Table 6 for the Approximate Sensitivity of some metals in AAS)
43
Element
Approximate
Element
Sensitivity (ppm)
Approximate Sensitivity (ppm)
As
0.50
Mg
0.01
Ca
0.10
Ni
0.15
Cd
0.05
K
0.05
Co
0.20
Ag
0.10
Cr
0.25
Na
0.05
Cu
0.10
Sn
5.00
Fe
0.15
Zn
0.05
Pb
0.50
Table 6. Approximate sensitivity, expressed in ppm, for several elements that may be analyzed by flame
44 APPENDIX G DOCUMENTATION
Watering of grasses with distilled water
Carabao and
Bermuda grass(ongoing
experiment)
45 CONTROL: Bermuda grass
Amended Samples: Bermuda and Carabao grass
46
Preparation and Filtering of the Samples ready for Reading
47
APPENDIX H Thesis Expenditures
Distilled water
400
Grasses (Bermuda & Carabao)
250
Gloves
50
Spray
50
Transportation
300
Lead Analysis
5,400
Printing & Bookbinding
500 ______________
TOTAL EXPENSES
Php 6,950
48 APPENDIX I RAW DATA of GROWTH RATE CARABAO GRASS (Leaf size) CARABAO GRASS 1 week 2nd week 3rd week (in cm) 55 55 55 55 68 98 55 65 76 (in cm) 60 60 60 60 79 98.5 60 68.9 79 (in cm) 55 55 55 55 76.6 99.7 st
TRIAL 1 Control 3000 ppm 6000 ppm TRIAL 2 Control 3000 ppm 6000 ppm TRIAL 3 Control 3000 ppm
4th week 55 105 85 60 125 89 55 115
6000 ppm
55
58.4
67.4
75
BERMUDA GRASS (Diameter)
1st week TRIAL 1 Control 3000 ppm 6000 ppm TRIAL 2 Control 3000 ppm 6000 ppm TRIAL 3 Control 3000 ppm 6000 ppm
190 190 190 198 198 198 197 197 197
BERMUDA GRASS 2nd week 3rd week (in cm) 190 190 160 144 160 144 (in cm) 198 198 210.4 236 219.6 243 (in cm) 197 197 210.8 200.3 200.9 220 49 APPENDIX J
RESULT of ANALYSIS through AAS (Carabao Grass)
4th week 190 146 145 198 250 250 197 220 240
50 APPENDIX K RESULT of ANALYSIS through AAS (Bermuda Grass)
51 Hannie Lou Faisan Anoc Tabok,Mandaue City,Cebu
[email protected] 09423483079/09106429083 PERSONAL INFORMATION Date of Birth Civil Status Nationality Religion
: : : :
March 7, 1993 Single Filipino Roman Catholic EDUCATIONAL ATTAINMENT
College
:
BS Chemistry-Physics Cebu Normal University Osmeña Blvd.,Cebu City
Secondary
:
Mandaue City Comprehensive National High School
Plaridel St.,Reclamation Area,Mandaue City ,Cebu JOB EXPERIENCES Student Assistant On-the-Job Trainee
Physics Laboratory-Cebu Normal University June 2011-March 2013 Department of Agriculture
May 2011, Mandaue Experiment Station
o Soils Laboratory o Pesticide Analytical Laboratory May 2012, M.Velez, Cebu City o Regional Feeds Laboratory POSITIONS HELD
Class Secretary, BS Chemistry-Physics – Cebu Normal University Batch 2009-2013 Member,Association of Student Assistants – Cebu Normal University 2011-2013
52 ORGANIZATIONS INVOLVED Council of Liberal Arts & Sciences (C.L.A.S.S.) – Cebu Normal University Junior Physics and Chemistry Society (JPACS) – Cebu Normal University Lector’s Ministry – National Shrine of St.Joseph
53 Sherlice Quiros Rom Cogon,Maslog,Danao City,Cebu
[email protected] 09106488759/09323013004 PERSONAL INFORMATION Date of Birth Civil Status Nationality Religion
: : : :
December 2, 1993 Single Filipino Roman Catholic EDUCATIONAL ATTAINMENT
College
:
BS Chemistry-Physics Cebu Normal University Osmeña Blvd.,Cebu City
Secondary
:
Compostela National High School
Poblacion,Compostela,Cebu JOB EXPERIENCES Student Assistant,Physics Laboratory-Cebu Normal University November 2009-October 2011 Student Assistant,Chemistry Laboratory-Cebu Normal University November 2011-March 2013 On-the-Job Trainee –Cebu Provincial Capitol Laboratories May 2011 o Provincial Engineering’s Laboratory o Provincial Agriculture’s Laboratory o Provincial Veterinary’s Laboratory o Provincial Water Analysis Laboratory On-the-Job Trainee- Regional Feeds Laboratory May 2012
54 POSITIONS HELD Alumni President,Batch 2009 - Compostela National High School 2011-Present Class Mayor – Cebu Normal University 2009-2011 Treasurer,Junior Physics and Chemistry Society- Cebu Normal University 2011-2013 Member,Association of Student Assistants – Cebu Normal University 2009-2013
ORGANIZATIONS INVOLVED Council of Liberal Arts & Sciences (C.L.A.S.S.) – Cebu Normal University Junior Physics and Chemistry Society (JPACS) – Cebu Normal University
Parish Pastoral Youth Council (PPYC) – St. Francis of Assisi Parish
55
