AUTHORS MWANSA MUKUKA DR. CHISHALA BENSON H. DR. MUTITI SAMUEL DR. UCHIDA YOSHITAKA APPSA CONFERENCE 6 TH NOVEMBER, 2017

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EVALUATION OF THE PHYTOREMEDIATION POTENTIAL OF LEMON GRASS (Cymbopogon citratus) AND VETIVER GRASS (Chrysopogon zizanioides) IN LEAD CONTAMINATED SOILS AUTHORS MWANSA MUKUKA DR. CHISHALA BENSON H. DR. MUTITI SAMUEL DR. UCHIDA YOSHITAKA APPSA CONFERENCE 6 TH NOVEMBER, 2017

Presentation Outline Introduction Statement of the problem Justification Study objectives Study hypotheses Materials and methods Results and discussion Conclusion Recommendations Acknowledgements References 2

Introduction The Lead (Pb) mining activities in Kabwe started at the beginning of the 20 th century and continued for 90 years until 1994. These mining activities left a lot of heavy metal contamination in Kabwe between 600 ppm and 27000 ppm within 2 km of the mine. The heavy metal contamination of the soils near and adjacent the Pb mine tailings arose from; the physical dumping of the mining wastes and, a large variety of industrial fumes with high heavy metals when they precipitated on the soil surface. 3

Introduction Mine metal tailings have a high concentration of toxic elements. These usually inhibit plant growth as most plants fail to withstand the toxicity. As such, the tailings are mostly bare. These tailings are prone to erosion as their top soils develop on unstable materials with low aggregation. An appropriate vegetation cover may reduce the erosion and immobilize toxic metals. 4

Statement of the Problem Heavy metals are a significant category of industrial pollutants due to their unique characteristics. The challenge of Pb contaminations in soil, also extends to include; How to sustainably and cost effectively, remove the heavy metal contaminants. How to dispose of the extracted heavy metals safely and, Pb tolerant plants to use for revegetation of Pb contaminated soils. Most of the known phytoremediators are expensive to manage. They have a lot of nutritional, water and re-planting requirements e.g. Chinese cabbage and sunflower. 5

Justification Vetiver and lemon grass are able to grow and thrive in many conditions inclusive of floods and droughts. These grasses do not necessarily require re-planting every year and are able to regain their growth vigor with rainfall once established. And since they re not directly edible, it poses no significant threat for re-introduction into the active environment. Vetiver grass is mostly used for soil erosion controls and soil stability exercises. However, little work has been done in Zambia to assess its phytoremediation potential for use in Pb contaminated soils. 6

Study Objectives Main Objective To evaluate the growth response and Pb uptake of vetiver and lemon grass in Pb contaminated soil. Specific Objectives To determine the uptake and distribution of Pb in vetiver grass. To determine the uptake and distribution of Pb in lemon grass. To evaluate the growth response of vetiver grass to Pb contamination. To evaluate the growth response of lemon grass to Pb contamination. 7

Materials and Methods Figure 1: The soil sampling sites (Kabwe, 2016) 8

Materials and Methods Study Site; The experiment was done at the University of Zambia, Great East Road Campus, School of Agricultural Sciences, Soil Science Department; under greenhouse conditions. The soil samples were collected from Kabwe. 9

Materials and Methods Table 1: Standard methods of Analysis PARAMETER METHOD (SOIL) METHOD(PLANTS) ph 0.01MCaCl2 N/A Electrical Conductivity Potentiometric N/A Ca, Mg, Na, K Ammonium Acetate Dry ashing Total Nitrogen Kjeldahl Kjeldahl Phosphorous Bray 1 Bray 1 Cu, Zn, Fe, Pb DTPA Dry ashing Organic matter Walkley % Black N/A Total Pb Aqua Regia N/A Bulk density Core ring N/A Texture Hydrometer N/A Standard methods were used to analyze the physical-chemical properties of the soil using the Analyst PerkinElmer 400-AAS. 10

Materials and Methods The vetiver and lemon grass (obtained from Chongwe area), was cut to a uniform height of 13 cm and 10 cm before planting. The treatments were replicated four times with each pot having 7 Kg of soil. The plants were grown for 120 days. 11

Materials and Methods The experimental design was Randomized Complete Block Design (RCBD). The data collected was analyzed using Analysis of Variance (ANOVA) to determine the effect of Pb on the plants. All tests were done at 95% confidence interval (or P 0.05 for significant difference). Mean separation was done using Duncan s Multiple range test. The GenStat version 18 software package was used in the analysis. 12

Results and Discussion Table 2: General properties of the Pb and Zn mine tailings PROPERTY CONTROL SITE 1 SITE 2 SITE 3 SITE 4 SITE 5 Soil texture Loam Sandy loam Sandy loam Loamy sand Loamy sand Loamy sand Bulk density (g/cm 3 ) 1.4 1.1 1.4 1 1 1 ph 4.76 4.82 5.15 4.73 4.52 5.07 EC (ms/cm) 0.10 0.08 0.06 0.55 0.13 0.27 CEC (cmol/kg) 1.29 4.75 3.57 7.71 5.08 2.56 ESP 1.51 5.21 5.41 2.05 4.94 4.75 %O.M. 2.76 0.37 2.21 1.39 1.37 0.73 Pb total (mg/kg) 0.00 3223.30 2701.70 30966.00 5823.30 20773.00 Pb DTPA (mg/kg) 0.00 865.50 720.50 1587.70 1510.00 1551.80 %N total 0.66 0.45 0.39 0.35 0.21 0.27 P (mg/kg) 18.70 1.46 16.57 1.02 0.83 1.40 K (cmol/kg) 0.76 0.46 0.73 0.30 0.37 0.24 Ca (cmol/kg) 0.32 3.67 2.02 6.63 4.21 1.89 Mg (cmol/kg) 0.19 0.34 0.58 0.61 0.21 0.27 Ca: Mg ratio 1.70 10.77 3.46 10.87 20.15 7.02 Zn (mg/kg) 1.63 8.47 12.53 14.75 12.53 16.20 Fe (mg/kg) 10.74 6.23 2.62 3.97 6.03 4.20 Cu (mg/kg) 1.39 1.67 3.07 12.43 12.35 12.02 13

Results and Discussion Site DTPA extractable Pb in soil (mg/kg) Table 4: Lead content of vetiver grass and lemon grass Vetiver grass (%Pb) shoots Vetiver grass (%Pb) roots Lemon grass (%Pb) shoots Lemon grass (%Pb) roots Control 0.00 e 0.0 i 0.0 p 0.00 f 0.0 p Site 1 865.50 d 0.02 f 0.10 q 0.00 f 0.30 q Site 2 720.50 c 0.02 f 0.06 r 0.02 g 0.01 p Site 3 1587.70 a 0.29 h 0.46 n 0.13 h 0.73 n Site 4 1510.00 a 0.08 g 0.21 m 0.02 g 0.22 m Site 5 1551.80 b 0.11 j 0.41 k 0.04 j 0.64 k Mean 0.10 0.25 0.04 0.38 Note: Values in the same column with the same superscripts are not statistically different at 95% C.I. (P > 0.05) 14

Results and Discussion Figure 2: Pb uptake by vetiver and lemon grasses 15

Results and Discussion Table 5: Pb distribution efficiency in vetiver and lemon grass Vetiver grass Lemon grass Sample Site TF BAF BCF TF BAF BCF control 0 0 0 0 0 0 Site 1 0.21 0.24 1.14 0.02 0.06 3.51 Site 2 0.37 0.29 0.77 3.46 0.30 0.09 Site 3 0.63 1.81 2.88 0.18 0.82 4.59 Site 4 0.38 0.53 1.38 0.08 0.11 1.46 Site 5 0.27 0.71 2.64 0.06 0.24 4.13 Mean 0.37 0.72 1.76 0.76 0.31 2.76 16

Results and Discussion Figure 3: Distribution of Pb in vetiver and lemon grass 17

Results and Discussion Table 6: General nutritional content of lemon grass and vetiver grass CONTROL SITE 1 SITE 2 SITE 3 SITE 4 SITE 5 LG VG LG VG LG VG LG VG LG VG LG VG %N total 0.14 f 0.09 a 0.09 g 0.04 e 0.11 i 0.03 b 0.06 h 0.05 c 0.09 g 0.07 d 0.09 g 0.07 d %P %K %Ca %Mg 0.12 a 0.36 e 0.10 b 0.07 f 0.09 c 0.08 g 0.10 b 0.07 h 0.06 d 0.07 h 0.06 d 0.05 i 1.23 f 1.50 j 1.09 h 1.41 j 0.96 i 1.43 j 1.15 g 1.06 k 1.11 gh 1.01 k 0.91 i 1.01 k 1.98 j 0.90 n 6.10 g 2.94 km 3.88 h 2.90 km 3.82 h 3.47 k 4.00 h 2.64 m 1.98 i 2.91 km 0.60 a 0.42 i 0.53 b 0.48 h 0.60 a 0.62 f 0.39 c 0.47 h 0.53 b 0.37 j 0.50 b 0.57 g %Zn 8.86E-03 k 7.90E-03 r 1.00E-02 j 2.91E-02 q 4.75E-02 g 6.99E-02 m 6.05E-02 f 5.36E-02 p 4.01E-02 h 6.28E-02 n 4.97E-02 g 5.44E-02 p %Fe 3.03E-02 m 5.42E-02 t 1.06E-01 k 3.23E-01 q 1.10E-01 jk 2.79E-01 s 2.42E-01 g 3.09E-01 r 1.66E-01 h 3.30E-01 p 1.35E-01 j 3.43E-01 n %Cu 4.15E-03 f 2.10E-03 p 1.09E-03 i 1.39E-03 q 3.99E-03 f 1.75E-03 r 1.42E-03 h 3.55E-03 n 1.94E-03 g 5.02E-03 m 8.13E-04 j 8.88E-03 j NOTE: Values with the same superscripts in the same row are statistically the same at 95% confidence interval (C.I.) (P > 0.05) LG = Lemon grass VG = Vetiver grass E = X 10^(n); where n is the stated number 18

Results and Discussion Figure 4: Biomass yield of vetiver and lemon grass in soil Pb 19

Results and Discussion Figure 5: Vetiver grass shoot at harvest Figure 6: Lemon grass shoot at harvest 20

Results and Discussion Figure 7: Vetiver grass roots at harvest Figure 8: Lemon grass roots at harvest 21

Conclusions The study established that both vetiver and lemon grass can grow in Pb contaminated soils. There was a decrease in biomass yield in vetiver and lemon grass with the increase in the Pb in the soil. The grasses were able to produce a substantial amount of root system despite the high levels of Pb, low ph, low O.M. content and low soil nutrition. 22

Conclusions Both lemon grass and vetiver grass took up Pb. Both lemon and vetiver grasses accumulated more Pb in the roots than the shoots (BAF < 1, BCF > 1). However, lemon grass translocated more Pb to the shoots than did vetiver grass (TF lemon grass > TF vetiver). Lemon grass accumulated an average of 0.38% Pb in the roots, and 0.04% Pb in the shoot. Vetiver grass accumulated an average of 0.25% Pb in the roots and 0.10% Pb in the shoots. Both lemon and vetiver grass can be used for phytostabilisation and revegetation. 23

Recommendation The ability to withstand the Pb toxicity and grow in the low fertility soil environment of the Pb tailing soils, makes vetiver and lemon grass a viable option for revegetation of the said place. With its fibrous and deep reaching root system, vetiver will be able to retain moisture in the soil, which may, in turn promote microbial and vegetative growth. The high amounts of Pb it accumulated in the roots (BCF > 1) by both vetiver and lemon grass also entails their ability to phytostabilise the Pb-contaminated mine tailings. Moreover, because both plants are not directly edible and may not necessarily require nutritional supplementation, they may be the sustainable green solution for the Pb/ Zn bare mine tailing of Kabwe and others. 24

Acknowledgements JICA and APPSA for the material and financial support. 25

THE END THANK YOU ALL FOR YOUR TIME AND ATTENTION 26