(Luận văn) toxicity test of several plant extracts as environmental friendly method to control mole cricket (gryllotalpa gryllotalpa linnaeus)

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY DO MANH DUNG lu an n va ie gh tn to TOXICITY TEST OF SEVERAL PLANT EXTRACTS AS ENVIRONMENTAL-FRIENDLY METHOD TO CONTROL MOLE CRICKET (GRYLLOTALPA GRYLLOTALPA LINNAEUS) p BACHELOR THESIS d oa nl w 2012-2016 z at nh oi Batch: International Training and Development Center lm ul Faculty: Environmental Science and Management nf va Major: Full-time an lu Study Mode: z m co l gm @ an Lu Thai Nguyen, 15/09/2016 n va ac th si DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program: Bachelor of Environmental Science and Management Student name: Do Manh Dung Student ID: DTN 1253180050 Thesis Title: Toxicity Test of Several Plant Extracts as Environmentalfriendly Method to Control Mole Cricket (Gryllotalpa lu gryllotalpa Linnaeus) an Dr Arinafril Naalim n va Supervisor (s): p ie gh tn to Ph.D Do Ngoc Son Abstract: In this research, we will observe the impact of Bio-pesticides and Chemical w oa nl pesticide: neem leaves, duku peels, durian peels, rambutan peels, Cypermethrin 50g/l d 50EC on crickets and finding which one is the best to compare with the (rest under an lu laboratory conditions) Methods used to this research is random design group, which nf va lm ul consisted of treatments, control, and repeating By making different stock z at nh oi solution from 25% to 5% and using a piece of cabbage then deep them inside the stock solutions in 20 seconds, after that give them to crickets for food With Cypermethrin z 50g/l 50EC, the concentration will be 2ml/l After research, Chemical pesticide is the @ l gm best one but it affects to human health, soil, plants, and environment With Bio- m co pesticides, plant extract from Durian peels is the best, after days almost crickets were died after day an Lu dead Especially, with plant extract from durian peels, stock solution is 25%, crickets n va ac th i si Key-words: Bio-pesticides, Cypermethrin, Mole Cricket, Neem Leaf, Rambutan Peel, Duku Peel, Durian Peel Number of pages: 33 pages Date of submission: 15/09/2016 Supervisor’s signature lu an n va p ie gh tn to d oa nl w nf va an lu z at nh oi lm ul z m co l gm @ an Lu n va ac th ii si ACKNOWLEDGEMENT The special thanks goes to my supervisor, Dr Arinafril Naalim of Sriwijaya University, Indralaya, Indonesia The supervision and support that he gave truly helped the progression and smoothness of the internship I am also highly thankful to Ph.D Ho Ngoc Son for his supervision, encouragement, guidance, and advice in writing this thesis In addition, I would like to thanks Ph.D Mulawarman, Lab NEMATOLOGY, lu an Department of Plant Protection, Sriwijaya University for helping and providing me n va necessary equipment for weigh, identify, and run my research ie gh tn to During study in Palembang, Indonesia there were highly interesting and p educative It helped me in understanding appropriately how to control insects by using oa nl w Bio-pesticides I am highly thankful to all office members of Sriwijaya University who helped and guided me during my internship d an lu Besides, I want to thanks to Didi, Dede, Henrda, Umiya, and Irwan for advice, nf va lm ul helping me to finish my study z at nh oi Last but not least, I would like to thank my parents for always by my side, support, and encouragement me every time z l gm @ Do Manh Dung m co an Lu n va ac th iii si TABLE OF CONTENTS LIST OF FIGURES LIST OF TABLES PART I INTRODUCTION 1.1 Research rationale 1.2 Research’s objectives 1.3 Research questions and hypothesis .5 lu an n va 1.4 Limitation gh tn to PART II LITERATURE REVIEW p ie 2.1 Neem leaves 2.2 Duku peels .7 oa nl w 2.3 Durian peels d lu nf va an 2.4 Rambutan peels .7 lm ul 2.5 Cypermethrin .8 z at nh oi PART III METHODS .9 3.1 Material and equipment z gm @ 3.2 Time and place co l 3.3 Research methods 10 m 3.3.1 Crickets 10 an Lu 3.3.2 Cabbages .10 n va ac th iv si 3.3.3 Plant extracts 10 3.3.4 Observation 11 3.3.5 Data processing .12 PART IV RESULTS 13 4.1 Set – up of the Research Subject 13 4.2 The relationship between plant extracts and chemical pesticide 14 4.3 The relationship between mortality of bio-pesticides and doses 28 lu an PART V DISCUSSION AND CONCLUSION 30 n va gh tn to 5.1 Discussion 30 5.1.1 The activities of plant extracts could impact on crickets 30 p ie nl w 5.1.2 Plant extract from durian peels got the most effective on cricket 31 d oa 5.2 Conclusion 31 lu nf va an REFERENCES 32 z at nh oi lm ul z m co l gm @ an Lu n va ac th v si LIST OF FIGURES Figure 4.2.1: The relationship between dose and mortality in day 16 Figure 4.2.2: The relationship between dose and mortality in day 18 Figure 4.2.3: The relationship between dose and mortality in day 20 Figure 4.2.4: The relationship between dose and mortality in day 22 Figure 4.2.5: The relationship between dose and mortality in day 24 lu an Figure 4.2.6: The relationship between dose and mortality in day 26 n va p ie gh tn to Figure 4.2.7: The relationship between dose and mortality in day 27 d oa nl w nf va an lu z at nh oi lm ul z m co l gm @ an Lu n va ac th si LIST OF TABLES Table 4.1.1: Set-up of the Research Subject 13 lu , probit equation of plant extracts and chemical pesticide in day 14 Table 4.2.2: , probit equation of plant extracts and chemical pesticide in day 15 Table 4.2.3: , probit equation of plant extracts and chemical pesticide in day 17 Table 4.2.4: , probit equation of plant extracts and chemical pesticide in day 19 Table 4.2.5: , probit equation of plant extracts and chemical pesticide in day 21 an Table 4.2.1: n va tn to Table 4.2.6: , probit equation of plant extracts and chemical pesticide in day 23 ie gh , probit equation of plant extracts and chemical pesticide in day 25 p Table 4.2.7: , probit equation of plant extracts and chemical pesticide in day 27 d oa nl w Table 4.2.8: nf va an lu Table 4.3.2: HSD Difference between means and significance of pairwise comparisons from the mortality of bio-pesticides 29 z at nh oi lm ul z m co l gm @ an Lu n va ac th si PART I INTRODUCTION 1.1 Research rationale Insects dominates all terrestrial environments that support human life, insects are the most significant competitors of us for food, fiber, and other natural resources They affect directly on agriculture food production by chewing the leaves of crop plans, sucking out plant juices, boring within the roofs, stems or leaves, and spreading plan pathogens They feed on natural fibers, destroy wooden building materials, ruin stored lu an grain, and accelerate the process of decay They also have a profound impact on the n va health of humans and domestic animals by causing annoyance, inflicting bites and to gh tn stings, and transmitting disease The economic impact of insects is measured not only by the market value of products they destroy and the cost of damage they inflict but p ie w also by the money and resources expended on prevention and control of pest oa nl outbreaks Although dollar values for these losses are nearly impossible to calculate, d especially when they affect human health and welfare, economists generally agree that an lu insects consume or destroy around 10% of gross national product in large, nf va industrialized nations and up to 25% of gross national product in some developing z at nh oi lm ul countries (John, 2016) Crickets (also known as “mole cricket”) belong to order Orthoptera, family Gryllotalpidae, and genus Gryllotalpa This family consists of a group of insect with z @ mainly cylindrical bodies, round heads, and long antennae They have thin antennae, gm co l hind legs modified for jumping, three-jointed tarsal (foot) segments, and two slender m abdominal sensory appendages (called cerci) The two forewings are stiff and leathery, an Lu and the two long, membranous hind wings are used in flying Male crickets produce n va musical chirping sound by rubbing a scraper located on one forewing along a row of ac th si about 50 to 250 teeth on the opposite forewing The purpose of cricket songs is attraction the females, the courtship, or mating, song, which induces the female to copulate; and the fighting chirp, which repels other males Both sexes have highly sensitive organs on the forelegs for sound reception There is a direct relationship between the rate of cricket chirps and temperature, with the rate increasing with increasing temperature Female crickets lay eggs into soil or on the plant stems, sometimes causing serious plant damage (Huber, 1989) Crickets feed many field crops, seeds, pods, or leaves and stems of seeding lu Crops can be attacked at any stage Crops in heavier soils are at greatest risk Most an n va damage is caused by crickets already in the area at planting or by adults flying into tn to crops Significant damage may be caused by adults and nymphs feeding on leaves, p ie gh stems and pods When crickets are present in plague numbers, seedling crops can be thinned to the point where replanting is necessary (Berenbaum, 1996) oa nl w Bio-pesticides is shortcut word of “Biological Pesticides”, it’s including several d classes such as: Biochemical pesticides, Microbial pesticides, and Plant-Incorporated- lu nf va an Protectants (PIPs) In this case, we are using biochemical pesticides Plant extracts were likely the earliest agricultural Bio-pesticides, as history records that nicotine was lm ul used to control plum beetles as early as the 17th century Experiments involving z at nh oi biological controls for insect pests in agriculture date back as far as 1835, when Agostine Bassi demonstrated that white-muscadine fungus (Beauveria bassiana) could z gm @ be used to cause an infectious disease in silkworm Experiments with mineral oils as co l plant protectants were also reported in the 19th century During the rapid institutional m expansion of agricultural research during the early 20th century, an ever-growing an Lu number of studies and proposal for Bio-pesticides were developed (Francis Borgio J, n va Sahayaraj K and Alper Susurluk I, 2011) ac th si lu an n va p ie gh tn to oa nl w Figure 4.2.3: The relationship between dose and mortality in day d The figure 4.2.3 shows the relationship between dose and mortality of bio- an lu nf va pesticides Plant extract from durian peels have strongest active from day to day lm ul At the dose 5% and 10% all crickets were died Plant extract from neem leaves can kill z at nh oi crickets at the dose 5%, 10%, 15%, and 25%, after day crickets in cups which there were treated by neem leaves at the dose 5%, 10%, and 25% is died all Plan extract z from rambutan peels kill crickets at the dose 5%, 10%, and 25% After days, crickets gm @ in cup have dose 5% there were died all Plant extract from duku peels kill cricket at m co l every doses, but it is slower than the other an Lu n va ac th 20 si Table 4.2.5: , probit equation of plant extracts and chemical pesticide in day lu Neem leaves = 37.356 Y = -2.634 + 0.071X Duku peels = 41.248 Y = -2.908 + 0.071X Durian peels = 41.065 Y = -2.896 + 0.071X Rambutan peels = 35.069 Y = -2.473 + 0.071X Cypermethrin 50EC = -4.532 Y = 0.320 + 0.071X an n va and probit equation of bio-pesticides after days The gh tn to Table 4.2.5 shows of plant extract from rambutan peels is p ie of cypermethrin is -4.532 The nl w lowest to compare with bio-pesticides, 35.069 After that is neem leaves with 37.356, of plant extract from rambutan d oa and duku peels, durian peels are around 41 The lm ul In day 5, nf va durian peels were dead an lu peels lowest because almost crickets in cups of plant extracts from neem leaves and = 2.6% The relationship between dose and mortality is still weak z at nh oi z m co l gm @ an Lu n va ac th 21 si lu an n va p ie gh tn to Figure 4.2.4: The relationship between dose and mortality in day d oa nl w an lu Looking at figure 4.2.4 we can see that, at the dose 20% of plant extract from nf va durian peels have dead crickets After days, all crickets in cups have plant extract lm ul from durian peels were dead, excepted at the dose 0% The mortality in cups of plant z at nh oi extract from rambutan peels is at the dose 0%, 10%, 15%, and 20%, after days, crickets in cup have dose 10% was dead all At the dose 0%, 15%, and 20% of plant z m co l gm @ extract from neem leaves have dead crickets Duku peels are at the dose 0% and 5% an Lu n va ac th 22 si Table 4.2.6: , probit equation of plant extracts and chemical pesticide in day Neem leaves = 25.714 Y = -3.507 + 0.136X Duku peels = 28.150 Y = -3.840 + 0.136X Durian peels = 38.026 Y = -5.187 + 0.136X Rambutan peels = 32.152 Y = -4.385 + 0.136X Cypermethrin 50EC = 0.749 Y = -0.102 + 0.136X lu an n va of cypermethrin is 0.749 The of tn to As depicted from the table 4.2.6, the ie gh neem leaves is 25.714, duku peels is 28.150, durian peels is 38.026, and rambutan of neem leaves is lowest to compare with bio- p peels is 32.152 So, in day 6, the oa = 0.2% The relationship between dose and mortality is weak d In day 6, nl w pesticides nf va an lu z at nh oi lm ul z m co l gm @ an Lu n va ac th 23 si lu an n va gh tn to Figure 4.2.5: The relationship between dose and mortality in day p ie w oa nl Figure 4.2.5 shows that, the mortality of neem leaves in day at the dose 15% d and 20% have dead crickets, after days, all cricket in cups of plant extract from neem an lu nf va leaves was all dead, excepted at the dose 0% The mortality at the dose 10% of plant lm ul extract from rambutan peels have active Plant extract from duku peels, at the dose crickets was dead z at nh oi 5%, 15%, 20%, and 25% have mortality, after days, in cups have dose 20% all z m co l gm @ an Lu n va ac th 24 si Table 4.2.7: , probit equation of plant extracts and chemical pesticide in day Neem leaves = 22.448 Y = -3.593 + 0.160X Duku peels = 27.102 Y = -4.339 + 0.160X Durian peels = 37.754 Y = -6.044 + 0.160X Rambutan peels = 32.350 Y = -5.179 + 0.160X Cypermethrin 50EC = 1.218 Y = -0.195 + 0.160X lu an n va of cypermethrin is 1.218 The of neem tn to The table 4.2.7 shows the ie gh leaves is 22.448, after that is duku peels, 27.102, rambutan peels is 32.350, and the last of durian peels, p one is durian peels is 37.754 After of neem leaves is lowest = 48.6% The relationship between dose and mortality is strong d In day 7, oa nl w to compare with bio-pesticides nf va an lu z at nh oi lm ul z m co l gm @ an Lu n va ac th 25 si lu an n va p ie gh tn to w d oa nl Figure 4.2.6: The relationship between dose and mortality in day an lu nf va The figure 4.2.6 shows relationship between dose and mortality of bio-pesticides lm ul in day At the dose 0% of plant extract from neem leaves have mortality Plant z at nh oi extract from rambutan peels at the dose 25% have dead crickets At the dose 5% and 25% of plant extract from duku peels have active, after days, all crickets living in the z cups have dose 25% is dead all m co l gm @ an Lu n va ac th 26 si Table 4.2.8: , probit equation of plant extracts and chemical pesticide in day Neem leaves = 32.942 Y = -6.963 + 0.211X Duku peels = 23.808 Y = -5.032 + 0.211X Durian peels = 32.239 Y = -6.814 + 0.211X Rambutan peels = 33.308 Y = -7.040 + 0.211X Cypermethrin 50EC = 2.255 Y = -0.477 + 0.211X lu an As depicted from the table 4.2.8, the of cypermethrin is 2.255 The = n va 23.808 belongs to plant extract from duku peels, lowest in day The rest are going tn to from 32.942 to 33.308 p ie gh d oa nl w nf va an lu z at nh oi lm ul z co l gm @ Figure 4.2.7: The relationship between dose and mortality in day m n va 10% and 15% an Lu Figure 4.2.7 shows the mortality of plant extract from duku peels is at the dose ac th 27 si 4.3 The relationship between mortality of bio-pesticides and doses Table 4.3.1: The relationship between mortality of bio-pesticides and doses Significant with Test of Bio-pesticides Significant with ANOVA Homogeneity of Variances Neem leaves 0.901 0.728 Duku peels 0.397 0.951 Durian peels 0.190 0.477 Rambutan peels 0.707 0.398 lu an The table 4.3.1 shows the relationship between mortality of plant extract from n va neem leaves with doses In the column Test of Homogeneity of Variances, significant tn to = 0.901 > 0.05 It has no difference variance between value of doses We have enough ie gh p condition to go to ANOVA column In ANOVA column, there is significant = 0.728 > oa nl w 0.05 It means there is no high significant difference between mortality of plant extract from neem leaves with doses d lu nf va an The table above shows that Significant in table Test of Homogeneity of Variances of duku peels is 0.397 > 0.05 It has no difference variance between value of lm ul doses We have enough condition to go to ANOVA column In ANOVA column, z at nh oi significant = 0.951 > 0.05 It means there is no high significant difference between mortality of plant extract from duku peels with doses z @ Looking at table 4.3.1, Significant = 0.190 > 0.05 from column Test of gm co l Homogeneity of Variances of durian peels It has no difference variance between value m of doses We have enough condition to go to ANOVA column In ANOVA column an Lu shows significant = 0.477 > 0.05 It means there is no significant difference between n va mortality of plant extract from durian peels with doses ac th 28 si As depicted from the table 4.3.1, Significant = 0.707 > 0.05 from table Test of Homogeneity of Variances of rambutan peels We have enough condition to go to ANOVA column In ANOVA table, significant = 0.398 > 0.05 It means there is no significant difference between mortality of plant extract from rambutan peels with doses Table 4.3.2: HSD Difference between means and significance of pairwise comparisons from the mortality of bio-pesticides Experiment Group 3.625 3.5 3.375 3.125 30 0.00 0.125 ns 0.255 ns 0.50 ns 26.375 ** 0.00 0.125 ns 0.375 ns 26.5 0.00 0.250 ns 26.625 ** 0.00 26.875 ** lu an n va tn to = 3.625 ie gh = 3.5 p = 3.375 0.00 d = 30 oa nl w = 3.125 ** nf va an lu lm ul Differences larger than 4.35 are significant at the α = 0.05 level are indicated indicated with ** z at nh oi with *, differences larger than 4.50 are significant at the α = 0.01 level are z The differences and significance of all pairwise comparisons are show in table @ l gm 3.3 The simultaneous pairwise comparisons indicate that the differences between biopesticides are not significant, and all the pairwise with cypermethrin are significantly m co different an Lu n va ac th 29 si PART V DISCUSSION AND CONCLUSION 5.1 Discussion 5.1.1 The activities of plant extracts could impact on crickets In the present study crickets were treated by cypermethrin 50EC of cypermethrin 50EC is always lowest from day to day 8, between 2.217 to -12.413 to compare with bio-pesticides It has strong impact on crickets in just only 30 minutes, all crickets in cups which treated by cypermethrin is dead all lu an In the present study crickets were treated by plant extracts from neem leaves, n va duku peels, durian peels, and rambutan peels In days from beginning, plant extract to gh tn from durian peels have higher impact than plant extracts from neem leaves, duku of plant extract from durian peels from day to day p ie peels, and rambutan peels is 50.972, 36.414, and 36.200 lower than the other plant extracts in the same day After nl w d oa day 5, all crickets in cup which treated by plant extract from durian peels is dead, an lu excepted in cup had stock solution 0% Plant extract from neem leaves had effect just of plant extract from neem leaves is 25.714 these is nf va after durian peels In day 6, lm ul lowest to the rest of bio-pesticides, and in the same day, all crickets in cups which z at nh oi treated by plant extract from neem leaves is dead, excepted in cup had stock solution 0% Plant extract from duku peels is the next In day and day 7, of it just after z gm @ plant extract from duku peels, it is 28.150 for day and 27.102 for day In day 8, co l crickets in cups have stock solution 10%, 15%, 20%, 25% are dead all Plant extract from rambutan peels is weakest to compare with the rest of bio-pesticides From day m an Lu to day 8, of rambutan peels are always highest In day 5, cricket in cups have n va stock solution 5% and 15% are dead all, the rest has still alive crickets ac th 30 si In day 7, = 48.6% Highest in days It means after days, bio-pesticides had strongest effects on crickets And by using HSD method to compare the difference between means and significance of pairwise comparisons from the mortality of biopesticides Those are no significant between bio-pesticides and significantly different between cypermethrin 50EC with each plant extracts 5.1.2 Plant extract from durian peels got the most effective on cricket Plant extract from durian peels got highest effect on cricket in plant extracts sector After days all of cricket was died in any stock solutions from 5% to 25% lu an Plant extract from neem leaves is days Plant extract from duku peels in days has n va alive cricket in stock solution 5% Plant extract from rambutan peels in days has tn to alive cricket in stock solution 10%, 20%, and 25% gh p ie 5.2 Conclusion nl w Cypermethrin was used in this experiment is the best to compare with plant d oa extract from neem leaves, duku peels, durian peels, rambutan peels It had strongly lu nf va an effect on cricket in very short time But cypermethrin is pesticide, when we use it for cricket on the fields, as a result, farmers and the lands can get cypermethrin’s lm ul disadvantages Thus, it needs to consider when farmers are using z at nh oi z m co l gm @ an Lu n va ac th 31 si REFERENCES Agency, U S (1997) Office of Pesticide Programs reference dose tracking report Washington, D.C Retrieved from http://ace.orst.edu/info/nptn/ tracking/tracking.htm Berenbaum, M R (1996) Bugs in the System: Insects and Their Impact on Human Affairs Basic Books, 44-45 Cantalamessa, F (1993) Acute toxicity of two pyrethroids, permethrin and lu an cypermethrin, in neonatal and adult rats Archives of Toxicology, 67, 510-513 n va Council, N R (1992) Neem: A Tree for Solving Global Problems Washington, DC: gh tn to National Academy Press p ie Council, N R (2000) The Future Role of Pesticides in US Agriculture Washington, nl w DC: National Academy Press d oa Dictionary, C O (2013) Neem Third Edition 2008 reprinted with corrections 2013 an lu (p 679) OXford, England: Oxford University Press nf va Francis Borgio J, Sahayaraj K and Alper Susurluk I (2011) Microbial Insecticides: lm ul Principles and Applications New York, USA: Nova Press z at nh oi Francis Borgio J; Sahayaraj K and Alper Susurluk I (2001) Microbial Insecticides: Principles and Applications New York, USA: Nova Press z gm @ Heaton, D D (2006) A Consumers Guide on World Fruit BookSurge Publishing Henry Yule, & Burnell, A C (1996) Neem The Anglo-Indian Dictionary (p 622) m co l Wordsworth Reference an Lu Huber, F (1989) Cricket Behavior and Neurobiology Cornell University Press n va ac th 32 si Jan , v S., & Cornelis , G G (1987) Flora, untuk sekolah di Indonesia[Flora, For Indonesian Schools] Jakarta,IN: PT Pradnya Paramita John, M (2016, March 24) Impact of Insects Retrieved from NC State University: https://projects.ncsu.edu/cals/course/ent425/library/tutorials/importance_of_ins ects/impact_of_insects.html Kiliỗ, A., & Akay, M T (2008) A three generation study with genetically modified Bt corn in rats: Biochemical and histopathological investigation Food and Chemical Toxicology, 46(3), 1164–1170 doi:10.1016/j.fct.2007.11.016 lu an Lawrence, J L., & Casida, J E (1982) Pyrethroid toxicology mouse intracerebral va n structure-toxicityrelationships Pestic Biochem Physiol, 18, 914 tn to Mordue, A J., & Blackwell, A (1993) Azadirachtin: an update Journal of Insect gh p ie Physiology, 39(11), 903-924 doi:10.1016/0022-1910(93)90001-8 nl w Morton , J F (1987) Rambutan, In: Fruits of warm climates Center for New Crops & d oa Plant Products, 262-265 lu nf va an Organization, W H (1989) Environmental Health Criteria Cypermethrin (Vol 82) Geneva: United Nations Environmental Programme, the International Labour lm ul Organization, and the World Health Organization z at nh oi Otte, D (2007) Australian Crickets (Orthoptera: Gryllidae) Academy of Natural Sciences, 17-24 z co l University Press gm @ Sapp, J (2003) Genesis: The Evolution of Biology (Vol Ch 7) New York: Oxford m Tindall, H D (1994) Rambutan Cultivation Food & Agriculture Org an Lu n va ac th 33 si Tomlin, C (Ed.) (1994) A World Compendium The Pesticide Manual Incorporating theagrochemicals handbook (10th ed.) Bungay, Suffolk, U.K: Crop Protection Publications Verheij, E.W.M; Coronel, R.E (1997) Sumber Daya Nabati Asia Tenggara 2: Buahbuahan yang dapat dimakan [Botanical Resources of South-East Asia 2] Jakarta: PROSEA – Gramedia lu an n va p ie gh tn to d oa nl w nf va an lu z at nh oi lm ul z m co l gm @ an Lu n va ac th 34 si