Isolation and detection of bacteria in soil, ash and sediment with the contamination of paraquat and glyphosate

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY NGUYEN DUC HAI TOPIC TITLE: ISOLATION AND DETECTION OF BACTERIA IN SOIL ASH AND SEDIMENT WITH THE CONTAMINATION OF PARAQUAT AND GLYPHOSATE BACHELOR THESIS Study Mode : Full - time Major : Environmental Science and Management Faculty : International Training and Development Center Batch : 2010 - 2015 Thai Nguyen, 23/01/ 2015 Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Environmental Science and Management Student Name Nguyen Duc Hai Student ID DTN1053110068 Thesis Title “Isolation and Detection of bacteria in soil, ash and sediment with the contamination of Paraquat and Glyphosate” Supervisor (s) Prof Chiu-Chung Young, National Chung Hsing University - Taiwan MSc Nguyen Huu Tho Abstract: Paraquat and Glyphosate are well-known as two of the most widely used herbicides in agricultural activities and landscape turf management However, their impacts are extremely numerous and serious on both human health and the environment, especially the uncontrolled usage of these herbicides with high levels causes soil contamination Currently, PCR is an useful and powerful technique, and it has been developed for various applications, particularly for DNA isolation and detection of microbial organisms, such as bacteria or viruses.Paraquat and Glyphosate were added to the soil, ash and sediment to make herbicide mixtures in this study Bacteria lived in herbicide mixtures were isolated, and then were detected by a PCR detection method In this study, there were 24 types of bacteria that were isolated and found out their names, in which 12 species of bacteria could grow well in the herbicide environment The results indicated that bacteria are able to survive in the herbicide contaminated condition It also demonstrated that those bacteria have potentials to utilize herbicide as a sole carbon source for their survival, degrade herbicide toxic compounds Paraquat and Glyphosate, and reduce significant the level of Paraquat and Glyphosate toxicity in the soil, ash and sediment were polluted by the herbicides Keywords Paraquat , Glyphosate, Microbial DNA isolation Herbicide, Biodegradation, Microbe Number of Pages 40 pages Date of Submission 22/01/2015 i AKNOWLEDGEMENT This thesis was completed under international collaboration between the Thai Nguyen University of Agriculture and Forestry, Thai Nguyen - Viet Nam and the National Chung Hsing University – Taiwan in the graduate internship program academic year 2010 - 2015 During implementation and completion of the subject, I have received the help and support of Thai Nguyen University of Agriculture and Forestry and National Chung Hsing University, on this occasion I would like to thank you for the invaluable help First of all I would like to express deep gratitude and respect to Professor Chiu Chung-Yong, who has direct supervision and guidance I made this topic, and I would like to thank Master Nguyen Huu Tho as a science instructor, he spent a lot of time and effort in helping me complete this research! This study can not complete without the encouragement and support of many colleagues, friends and family members, Finally, I would like to thank everyone who helped me complete this study! Thai Nguyen, January 15th, 2015 Supervisor Nguyen Huu Tho Student Nguyen Duc Hai ii TABLE OF CONTENTS LIST OF FIGURES LIST OF TABLES LIST OF ABBREVIATIONS PART I INTRODUCTION .4 1.1 Research rationale .4 1.2 Research’s objectives 1.3 Research questions 1.4 Limitations 1.5 Definitions 1.5.1 Herbicide 1.5.2 Soil, ash and sediment .6 PART II LITERATURE REVIEW 2.1 Overview of Herbicide .7 2.1.1 Classification of herbicides 2.1.2 Effects of herbicide with environment and human 2.1.3 Paraquat 2.1.4 Glyphosate 2.2 Overview of soil, ash, sediment and soil pollution 10 2.2.1 Soil 10 2.2.2 Ash 11 2.2.3 Sediment 11 2.2.4 Soil Contamination 12 2.3 Polymerase Chain Reaction ( PCR ) 14 2.3.1 Definition 14 2.3.2 Process of PCR Method 14 2.3.3 Application of PCR 16 2.4 Related study 17 2.4.1 International researches 17 iii 2.4.2 Domestic research 18 PART III METHODS 19 3.1 Materials 19 3.1.1 Bushnell Haas basal medium 19 3.1.2 NA medium 19 3.1.3 Preparing Herbicide Toxicity 20 3.1.4 Microbial DNA isolation 20 3.1.5 Equipment 20 3.2 Methods 21 3.2.1 Preparing Nutrient Broth Agar medium and Bushnell Haas culture medium 21 3.2.2 Preparing Herbicide Toxicity 21 3.2.3 Serial dilution of herbicide mixtures 22 3.2.4 Streaking Plates 22 3.2.5 Microbial DNA isolation 22 3.2.6 Polymerase chain reaction (PCR) detection of bacteria 23 3.2.7 Imaging and Analysis 25 PART IV RESULTS 26 4.1 Preparing herbicide toxicity 26 4.2 Serial dilution of herbicide mixtures 26 4.3 Microbial DNA isolation 28 4.4 Test the growth of bacteria in contamination environment 34 PART V DISCUSSION AND CONCLUSION 37 5.1 DISCUSSION 37 5.2 CONCLUSION 37 REFERENCES 38 iv LIST OF FIGURES Figure 2.1 The molecular structure of Paraquat Figure 2.2 The molecular structure of Glyphosate .9 Figure 2.3 The three main step of PCR process .16 Figure 3.1 PCR process for 16S rDNA .24 Figure 4.1.The color and density of bacteria in process of dilution for Ash-PRQ herbicide mixtures 29 Figure 4.2.The color and density of bacteria in process of dilution for Ash-GPT herbicide mixtures 29 Figure 4.3.The color and density of bacteria in process of dilution for .30 Sediment-PRQ herbicide mixtures .30 Figure 4.4.The color and density of bacteria in process of dilution for Sediment-GPT herbicide mixtures 30 Figure 4.5.The color and density of bacteria in process of dilution for Soil-PRQ herbicide mixtures 31 Figure 4.6.The color and density of bacteria in process of dilution for Soil-GPT herbicide mixtures 31 LIST OF TABLES Table 2.1 Classification for the size of sediment (Holmes, 1978) 12 Table 3.1 Content of BH medium 19 Table 3.2 Content of NA medium 19 Table 3.3 The content of PCR tube 24 Table 4.1 Herbicide mixtures using in this experiment 26 Table 4.2 The C.F.U of bacteria per 1mL in each type of herbicide mixtures 27 Table 4.3 Number of bacteria detected in six herbicide mixtures 28 Table 4.4 The density of each type of bacteria in Ash – PRQherbicide mixture 32 Table 4.5 The density of each type of bacteria in Ash - GPTherbicide mixture 32 Table 4.6 The density of bacteria in herbicide mixture 33 Table 4.7 Bacteria isolated in ash, soil and sediment with contamination of Paraquat and Glyphosate 34 Table 4.8 Ability for survival of bacteria in herbicide mixture .36 LIST OF ABBREVIATIONS PRQ Paraquat GPT Glyphosate C.F.U Colony Forming Unit PCR Polymerase chain reaction PART I INTRODUCTION 1.1 Research rationale At the end of the 20th century and the early of the 21st century, it is a period that marks the extremely strong development of science and technology in Vietnam and other nations in the world As an agricultural country with a long wet rice culture, Vietnam primarily takes advantages of available resources Nevertheless, by using outdated farming methods with the presence of plant protection products and the unscientific use of those ones, it causes contamination of the environment, especially in soil, water, sediments, etc in Vietnam At the present, there are thousands of types of pesticides, herbicides, soil improvement products used Each type of those requests users must have certain knowledge about the dosage and usages to have the best efficiency and less impact on the environment as well Nonetheless, in fact, almost of users measure the dose by the naked eyes and unscientific manners They use pesticides, herbicides or other products arbitrarily, thus the efficiency is impacted and the environment is received adverse effects from that (Sribanditmongkol et al., 2012) 1.2 Research’s objectives In this study, Paraquat and Glyphosate were added to the soil, ash and sediment for the enrichment and using a specific PCR detection method Bacteria that could grow in herbicide environment were detected That were demonstrate that those bacteria could utilize herbicide as a sole carbon source for their survival Therefore, this research could confirm the growth of bacteria in the that contaminated condition as well as found out the potential bacteria with a strong ability to remove Paraquat and Glyphosate compounds in the environment 1.3 Research questions Each bacterium has a different metabolic system to eliminate herbicide The application of bacteria in detected in this study was significant for bioremediation technology Therefore, this research answers the following questions How many kinds of bacteria have been isolated and detected in herbicide mixtures? What kinds of bacteria have ability to survive and grow in the environment contaminated by these herbicides? How is the growth of bacteria in the herbicide contaminated environment? 1.4 Limitations This study concentrated on the ability of bacteria to reduce Paraquat and Glyphosate concentration and their growth within the contaminated environment in laboratory This research brought many significances in scientific learning and research areas In terms of this aspect, doing the proposed research required the application and promotion of academic knowledge, and the results are useful materials or references for future researches The potential bacteria, were able to biodegrade or reduce Paraquat and Glyphosate concentration effectively, could apply in soil, ash or sediments which were contaminated by these kinds of herbicide Besides, this study was significant to bioremediation applications and treatment of the environmental issues 1.5 Definitions 1.5.1 Herbicide Herbicide is defined as weed killers or a pesticide used to kill weeds or unwanted wild plants which makes crops grow poorly and impact to crop yield and quality of agricultural products (Kellogg et al., 2000) PART IV RESULTS 4.1 Preparing herbicide toxicity After adding mL Herbicide (Paraquat and Glyphosate) and 10g environment (soil, ash and sediment) on the BH 50mL media, there were six types of herbicide mixtures, as shown on table 4.1 Table 4.1 Herbicide mixtures using in this experiment Categories Paraquat ( mL ) Glyphosate ( mL ) Soil ( 10 g ) Soil - PRQ Soil - GPT Ash ( 10 g ) Ash - PRQ Ash - GPT Sediment ( 10 g ) Sediment - PRQ Sediment - GPT 4.2 Serial dilution of herbicide mixtures For first times of dilution, the density of bacteria colonies in samples of herbicide mixtures were too high, thus the C.F.U number could not be measured However, after the 6th time or the 7th time of dilution, the C.F.U per mL of samples were determined in a suitable a range that is able to count The results showed in table 4.2 26 Table 4.2 The C.F.U of bacteria per 1mL in each type of herbicide mixtures Final Ash-PRQ Ash-GLP Sediment-PRQ Sediment-GLP Soil-PRQ Soil-GLP Number dilution of on Colonies CFU/mL Colonies CFU/mL Colonies CFU/mL Colonies CFU/mL Colonies CFU/mL Colonies CFU/mL dilute plate 10-1 -* - - - - - - - - - - - 10-2 - - - - - - - - - - - - 10-3 - - - - - - - - - - - - 10-4 - - - - - - - - - - - - 10-5 373 932,5 298 496,67 812 1353,33 187 311,67 443 738,33 518 863,33 10-6 98 294 48 96 510 1020 86 172 101 202 105 210 10-7 25 87,5 14 32,667 119 277,67 20 46,67 19 44,33 18 42 10-8 12 0 15 40 24 0 5,33 (*) – mean uncountable 27 4.3 Microbial DNA isolation Each herbicide mixture had single types of bacteria in table 4.3, thereby the total isolated bacteria were 24 types after seven days for incubation of six herbicide mixtures in seven days Table 4.3 Number of bacteria detected in six herbicide mixtures Type of herbicide mixture Number of Bacteria Ash - PRQ Ash - GPT Sediment - PRQ Sediment - GPT Soil - PRQ Soil - GPT The figure 4.1, 4.2, 4.3, 4.4, 4.5 and 4.6 showed how different in density of samples of herbicide mixtures (Ash-PRQ, Ash-GPT, Sediment-PRQ, Sediment-GPT, Soil-PRQ, Soil-GPT) among each time of dilution respectively As can be seen from all figures, the samples of herbicide mixtures diluted with the level of 10-1 and 10-2 had much more colonies of bacteria than those diluted with the level of 10-6 or 10-7 The bacteria colonies were decreased after dilution process was occurred 28 Figure 4.1.The color and density of bacteria in process of dilution for Ash-PRQ herbicide mixtures Figure 4.2.The color and density of bacteria in process of dilution for Ash-GPT herbicide mixtures 29 Figure 4.3.The color and density of bacteria in process of dilution for Sediment-PRQ herbicide mixtures Figure 4.4.The color and density of bacteria in process of dilution for SedimentGPT herbicide mixtures 30 Figure 4.5.The color and density of bacteria in process of dilution for Soil-PRQ herbicide mixtures Figure 4.6.The color and density of bacteria in process of dilution for Soil-GPT herbicide mixtures 31 Each bacterium in particular herbicide polluted conditions had different ability to grow In Ash - PRQ herbicide mixture, Luteimonas Aestuariiand Pseudomonas taiwanensishad high density, and were opposite to the others in the same condition, as shown on table 4.4 Table 4.4 The density of each type of bacteria in Ash – PRQherbicide mixture Name of bacteria Density Luteimonas Aestuarii 100 up to 150 CFU/mL Pseudomonas taiwanensis 100 up to 150 CFU/mL Phenylo bacterium koreense 30 up to 100 CFU/mL Shinellazoogloeoides 30 up to 100 CFU/mL Table 4.5 indicated that Ochrobactrum intermediumhas the highest density in in the environment of Ash - GPT herbicide mixture Table 4.5 The density of each type of bacteria in Ash - GPTherbicide mixture Name of bacteria Density Brachy bacterium para conglomeratum 100 up to 150 CFU/mL Ochrobactrum intermedium 150 up to 300 CFU/mL Brevundimonas nasdae 30 up to 100 CFU/mL Rhodococcusimtechensis 30 up to 100 CFU/mL The results in table 4.6 also showed that the density of bacteria were not equal among species of bacteria, even they received the same conditions The growth and tolerance of herbicide toxicity of each type of bacteria were not the same 32 Table 4.6 The density of bacteria in herbicide mixture Name of bacteria Density Thermomonas koreensis 150 up to 300 CFU/mL Ensife radhaerens 30 up to 100 CFU/mL Brevundimonas vesicularis 30 up to 100 CFU/mL Stenotrophomonas terrae 30 up to 100 CFU/mL Ochrobactrum ciceri 30 up to 100 CFU/mL Pseudomonas nitroreducens 30 up to 100 CFU/mL Pseudomonas geniculate 100 up to 150 CFU/mL Rhodanobacter xiangquanii 100 up to 150 CFU/mL Ochrobactrum lupine 30 up to 100 CFU/mL Stenotrophomonas maltophilia 100 up to 150 CFU/mL Pseudomonas plecoglossicida 100 up to 150 CFU/mL Alcaligenes faecalis 30 up to 100 CFU/mL Pseudomonas hibiscicola 150 up to 300 CFU/mL Alcaligenes aquatilis 30 up to 100 CFU/mL Thermomonas brevis 100 up to 150 CFU/mL Ochrobactrum tritici 30 up to 100 CFU/mL • Isolation and identifies bacteria The results in table 4.7 indicated that 24 species of bacteria were isolated and detected with specific scientific names These bacteria were analyzed and determined with high accuracies, more than 98% of accuracy in compared to known 33 microorganisms Especially, bacteria in the total of 24 bacteria were 100% of accuracy with bacteria that were discovered and published before Table 4.7 Bacteria isolated in ash, soil and sediment with contamination of Paraquat and Glyphosate Type Name of bacteria Ash - PRQ Ash - GPT Soil - PRQ Soil - GPT Sediment PRQ Sediment GPT No of similarity Percengate (%) Luteimonas Aestuarii 660/673 98.00 Pseudomonas taiwanensis 680/683 99.60 Phenylo bacterium koreense 519/525 98.90 Shinellazoogloeoides 628/630 99.70 Brachy bacterium para conglomeratum 575/575 100.00 Ochrobactrum intermedium 652/652 100.00 Brevundimonas nasdae 491/491 100.00 Rhodococcusimtechensis 629/634 99.20 Thermomonas koreensis 631/631 100.00 Ensife radhaerens 673/675 99.70 Brevundimonas vesicularis 634/638 99.40 Stenotrophomonas terrae 611/615 99.30 Ochrobactrum ciceri 684/686 99.70 Pseudomonas nitroreducens 676/676 100.00 Pseudomonas geniculate 665/665 100.00 Rhodanobacter xiangquanii 653/659 99.10 Ochrobactrum lupine 566/566 100.00 Stenotrophomonas maltophilia 703/705 99.70 Pseudomonas plecoglossicida 714/714 100.00 Alcaligenes faecalis 675/678 99.60 Pseudomonas hibiscicola 687/688 99.90 Alcaligenes aquatilis 679/682 99.50 Thermomonas brevis 677/688 98.40 Ochrobactrum tritici 661/661 100.00 4.4 Test the growth of bacteria in contamination environment As shown from the table 4.8, after 48 hours, all bacteria in Ash – GPT, Soil – PRQ, Sediment – GPT herbicide mixtures could survive and grow in the herbicidecontaminated environment, and just one type of bacteria in Soil – GPT is survival, namely 34 Ochrobactrum ciceri After 72 hours, only bacteriaPseudomonas hibiscicolain Sediment – GPT cannot continue to develop However, the other bacteria in Ash – PRQ, Sediment – PRQ, Soil – GPT herbicide mixtures could stand by toxicity of herbicide, thereby they were died If bacteria cannot grow in the presence of herbicides, they have inability to use herbicide as a carbon source for living The results indicates that the total bacteria that grew in the herbicide conditions are 12 bacteria, they can degrade Paraquat and Glyphosate, and utilized the herbicides as a carbon source Bacteria growing ( ++ ), stable ( + ) or die (- ) are showing in table 4.8 35 Table 4.8 Ability for survival of bacteria in herbicide mixture Ability Name of bacteria 48 hours 72 hours Luteimonas Aestuarii - - Pseudomonas taiwanensis - - Phenylo bacterium koreense - - Shinellazoogloeoides - - Brachy bacterium para conglomeratum ++ + Ochrobactrum intermedium ++ + Brevundimonas nasdae ++ + Rhodococcusimtechensis ++ + Thermomonas koreensis ++ ++ Ensife radhaerens + + Brevundimonas vesicularis ++ + Stenotrophomonas terrae ++ + Ochrobactrum ciceri + + Pseudomonas nitroreducens - - Pseudomonas geniculate - - Rhodanobacter xiangquanii - - Ochrobactrum lupine - - Stenotrophomonas maltophilia - - Pseudomonas plecoglossicida - - Alcaligenes faecalis - - Pseudomonas hibiscicola + - Alcaligenes aquatilis ++ + Thermomonas brevis ++ + Ochrobactrum tritici ++ + 36 PART V DISCUSSION AND CONCLUSION 5.1 DISCUSSION After seven days for incubation of six herbicide mixtures, 24 types of bacteria were detected with their scientific name by PCR The severity of herbicide contamination can be determined based on C.F.U per mL The higher C.F.U number is, the less level of herbicide toxicity is The results showed that 12 bacteria, that survived after 3days in the contaminated environment, have potentials to degrade and neutralize Paraquat and Glyphosate, and use these types of herbicide as carbon source for their growth 5.2 CONCLUSION - The density of bacteria were not equal among species of bacteria, even they received the same conditions It depends on potential of each type of bacteria in the growth and tolerance of Paraquat and Glyphosate toxicity - There were 24 species of bacteria that were successfully isolated and detected in herbicide mixtures with specific scientific names These bacteria were analyzed and determined with high accuracies in compared to known bacteria that were discovered and published before - Twelve bacteria had potential to survive and grow well in the contaminated environment by Paraquat and Glyphosate, namely Without carbon source, these bacteria had ability to degrade and neutralize Paraquat and Glyphosate, and use these types of herbicide as carbon source for their growth The twelve bacteria are very useful in treatment of herbicide contamination, such as soil pollution or water pollution if they are applied in microbial degradation of bacteria in further researches 37 REFERENCES Andrew, H.C and John, P.H.R (2011) Herbicides and Plant Physiology John Wiley & Sons Bartlett, J.M.S and Stirling, D (2003) History of the Polymerase Chain Reaction In: PCR Protocols Methods in Molecular Biology (2nd), pp 3–6 Buzik, S.C., Schiefer, H.B., and Irvine, D.G (1997) Understanding Toxicology: Chemicals, Their Benefits and Risks Boca Raton, pp 31 Cristina, B., Ivan I.P.B., and Kevin, R (2007) Biointerphases Chesworth, W.E (2008) Encyclopedia of soil science Dordrecht, Netherlands: Springer xxiv ISBN 1-4020-3994-8 Diaz, K.E.M., Mártire, D.O., Gonzalez, M.C., and Rosso, J.A (2010) Degradation of the herbicides clomazone, paraquat, and glyphosate by thermally activated peroxydisulfate J Agric Food Chem 2010 Dec 22 Dinis-Oliveira, R.J., Remião, F., Carmo, H., Duarte, J.A., Navarro, A S., Bastos, M.L., and Carvalho, F (2006) Paraquat exposure as an etiological factor of Parkinson's disease NeuroToxicology 27 (6),pp 1110–22 Franz, J.E (1974) N-phosphonomethyl-glycine phytotoxicant compositions Monsanto Company Gorell, J.M., Johnson, C.C., Rybicki, B.A., Peterson, E.L., and Richardson, R.J (1998) The risk of Parkinson's disease with exposure to pesticides, farming, well water, and rural living Neurology 50 (5), pp 1346–50 Havens, P.L., Sims, G.K., and Erhardt, Z.S (1995) Fate of herbicides in the environment Handbook of weed management systems M Dekker, pp 245-278 38 Holmes, G (1978), Sedimentary Environments: Processes, Facies and Stratigraphy Cambridge, MA: Blackwell Science ISBN 0-632-03627-3 Hwang, K.Y., Eun, Y.L., and Sae, Y.H (2002), Paraquat intoxication in Korea Archives of Environmental Health An International Journal 57.2, pp 162-166 IUPAC, (1997), Compendium of Chemical Terminology Gold Book, 2nd ed Jimenez, D.R., Marlene, V.P., and Carlos, V.P (2008) Paraquat apoptosis in human lymphocytes: protective and rescue effects of glucose, cannabinoids and insulinlike growth factor-1 Growth Factors 21, pp 49-60 Joseph, S., and David, W.R (2001) Molecular Cloning: A Laboratory Manual (3rd.) 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the diagnosis of 16 hereditary diseases or genetic disorders; the identification of genetic fingerprints; and the detection and diagnosis...1.5.2 Soil, ash and sediment 1.5.2.1 Soil Soil is a natural body, including solids (minerals and organic matter), liquids, and gases occurs on the surface, occupies space and is characterized by one or both of the following: leg sun, or the layer, which is distinguished from the original material as a result of the addition, losses, transfer, and transformation of energy and matter, or the ability... flasks, namely Paraquat- soil mixture, Paraquatash mixture, Paraquat -sediment mixture, Glyphosate- soil mixture, Glyphosate -ash mixture and Glyphosate -sediment mixture The flasks were carefully covered with rubber caps and stored in a plastic bag Then they were incubated at the shaking incubator 100 r.p.m for 1 week enrichment 21 3.2.3 Serial dilution of herbicide mixtures Each flask of herbicide-soil... 2010) The application of microbe in degradation of herbicide is still a new and potential approach Singh B, Singh K in India (2014) had applied microbe for of herbicides (Singh, 2014 ) 2.4.2 Domestic research Currently, the technology of using microbe to degrade or eliminate herbicide is quite new in Vietnam, thus there is less scientific research about treating Paraquat and Glyphosate in contaminated... amounts of initial material Other applications of PCR include DNA sequencing to determine unknown PCR-amplified sequences in which one of the amplification primers may be used in Sanger sequencing, isolation of a DNA sequence to expedite recombinant DNA technologies involving the insertion of a DNA sequence into a plasmid or the genetic material of another organism Bacterial colonies (E coli) can be rapidly... on human health and the environment, especially pollute the soil environment Today, researchers have lots of methods to reduce the influences of Paraquat and Glyphosate, and many other studies focus on the crops that are resistant to herbicide toxicity For example studied of Diaz et al, 2010 in 17 Paraguay shown the degradation of the herbicides clomazone, paraquat, and glyphosate by thermally activated... 700 µL of the supernatant was put into the Spin Filter and centrifuged at 10,000 x g for 30 seconds at room temperature The flow through was discarded, and the remaining supernatant was added more into the Spin Filter and continues to be centrifuged The next step was that 300 µL of solution MD4 was add more into the supernatant and also centrifuged at 10,000 x g for 30 seconds at room temperature The ... 4.4 The density of each type of bacteria in Ash – PRQherbicide mixture 32 Table 4.5 The density of each type of bacteria in Ash - GPTherbicide mixture 32 Table 4.6 The density of bacteria in. .. carbon source for their survival, degrade herbicide toxic compounds Paraquat and Glyphosate, and reduce significant the level of Paraquat and Glyphosate toxicity in the soil, ash and sediment were... bacteria in soil, ash and sediment with the contamination of Paraquat and Glyphosate Supervisor (s) Prof Chiu-Chung Young, National Chung Hsing University - Taiwan MSc Nguyen Huu Tho Abstract: Paraquat