PERFORMANCE OF EGG PARASITOID Trichogramma chilonis (Ishii) UNDER LABORATORY CONDITIONS

Transcription

1 PERFORMANCE OF EGG PARASITOID Trichogramma chilonis (Ishii) UNDER LABORATORY CONDITIONS By DILEEP, R. C. B.Sc. (Agri.) DEPARTMENT OF AGRICULTURAL ENTOMOLOGY, FACULTY OF AGRICULTURE, DR. BALASAHEB SAWANT KONKAN KRISHI VIDYAPEETH, DAPOLI , DIST. RATNAGIRI (M.S.) APRIL, 2012

2 PERFORMANCE OF EGG PARASITOID Trichogramma chilonis (Ishii) UNDER LABORATORY CONDITIONS A thesis submitted to the DR. BALASAHEB SAWANT KONKAN KRISHI VIDYAPEETH, DAPOLI (Agricultural University) Dist. Ratnagiri (Maharashtra State) in partial fulfilment of the requirements for the degree of MASTER OF SCIENCE (AGRICULTURE) In AGRICULTURAL ENTOMOLOGY By DILEEP, R. C. B.Sc. (Agri.) DEPARTMENT OF AGRICULTURAL ENTOMOLOGY, FACULTY OF AGRICULTURE, DR. BALASAHEB SAWANT KONKAN KRISHI VIDYAPEETH, DAPOLI , DIST. RATNAGIRI (M.S.) APRIL, 2012

3 32 Dr. (Mrs.) K. V. Naik M.Sc. (Agri.), Ph.D. : Chairman and Research Guide: Assistant Professor, Department of Agril. Entomology, (P. College D. Veerkar) of Agriculture, Dapoli. Assistant Dist. Professor, Ratnagiri (M.S.) PIN Department of Agricultural Economics, College of Agriculture, Dapoli : Members: (S. S. Wadkar) Associate Professor, Department of Agricultural Economics, College of Agriculture, Dapoli (A. C. Sawant) Professor, Department of Agronomy, College of Agriculture, Dapoli This is to certify that the thesis entitled Performance of egg parasitoid (V. G. Naik) Trichogramma chilonis (Ishii) under laboratory Assistant Professor, Department of Agricultural Economics, College of Agriculture, Dapoli C E R T I F I C A T E conditions submitted to the Faculty of Agriculture, Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, Dist. Ratnagiri, (Maharashtra State), in the partial fulfilment of the requirements for the degree of MASTER OF SCIENCE (AGRICULTURE) in AGRICULTURAL ENTOMOLOGY, embodies the results of a piece of bona-fide research carried out by Mr. DILEEP, R. C. under my guidance and supervision. No part of this thesis has been submitted for any other degree or diploma. All the LIST assistance OF TABLES. and help received during the course of investigation and the sources of literature have been duly acknowledged by him. Place: Dapoli Date: April, 2012 (K. V. Naik) Chairman Advisory Committee and Research Guide

4 33 PERFORMANCE OF EGG PARASITOID Trichogramma chilonis (Ishii) UNDER LABORATORY CONDITIONS A thesis submitted to the DR. BALASAHEB SAWANT KONKAN KRISHI VIDYAPEETH, DAPOLI (Agricultural University) Dist. Ratnagiri (Maharashtra State) in partial fulfilment of the requirements for the degree of MASTER OF SCIENCE (AGRICULTURE) In AGRICULTURAL ENTOMOLOGY By DILEEP, R. C. B.Sc. (Agri.) Approved by the Advisory Committee Chairman and Research Guide: (K. V. Naik) Assistant Professor, Department of Agricultural Entomology, College of Agriculture, Dapoli. Members: (S. K. Mehendale) Assistant Professor, Department of Agricultural Entomology, College of Agriculture, Dapoli. (M. S. Joshi) Assistant Professor, Department of Plant Pathology, College of Agriculture, Dapoli.

5 34 ACKNOWLEDGEMENT There are several occasions when you say Thanks to someone in your life time, but when a person divert your life towards a new achievement without whom you can t think about that, that condition creates a real respect and faith in your heart and your words become an Acknowledgement in respect to that great personality. At this time, I am on that golden moment of my life I would like to express my sincere gratitude to Dr. (Mrs.) K. V. Naik, Assistant Professor, Department of Agricultural Entomology, College of Agriculture, Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli. I express my deep and sincere gratitude to him for whose most valuable and inspirative guidance, keen interest, concrete suggestions, constant encouragement, enormous help and constructive criticism throughout my academic career and above all, playing an important role in moulding my personality. My profound and sincere thanks to the members of my advisory committee, Dr. S. K. Mehendale, Assiatant professor, Department of Agricultural entomology and Dr. M. S. Joshi, Assistant Professor, Department of Plant Pathology, College of Agriculture, Dapoli for their candid suggestions, assiduous and astute approach throughout the course of investigation and also for their ebullience in shaping out this manuscript. I wish to express my cordial thanks to Dr. A. L. Narangalkar, Head, Department of Agricultural Entomology, College of Agriculture, Dapoli for his co-operation and assistance during the course of investigation. I am personally indebted to Dr. P. D. Patil, Ex. Head, Department of Agril. Entomology, Dr. S. K. Godase, Associate Professor, Dr. V. S. Desai, Assistant Professor, Prof. M. S. Karmarkar, Assistant Professor, Prof. S. D. Desai, Assistant Professor, Department of Agril. Entomology, College of Agriculture, Dapoli and Prof. R. S. Patil, Scientist, AICRP, College of forestry, Dapoli for their love, help and guidance during the period of this study. I place on record my cordial thanks to, Hon ble Dr. K. E. Lavande, Vice Chancellor, Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dr. G. D. Joshi, Ex. Dean, Faculty of Agriculture, college of Agriculture, Dr. C. V. Bhambure, Dean, Faculty of Agriculture and Associate Dean, college of Agriculture, Dapoli for their valuable guidance and provision of necessary facilities for conducting this study. I would like to thank Mr. R. H. Mhatre, Agril. Assistant, Mr. Awati, Field Assistant, Mr. Pavse,Clerk and Rahate Mavshi, Rale kaka, Bhambid kaka, Viju kaka, Dhamane kaka, Pagade Kaki and all staff members who helped me regularly by making all the material available at hand very promptly whenever needed. I am sincerely thankful to The Director, NBAII, Bengaluru, for providing Trichogramma and Corcyra culture. I have no words to express my special thanks to Dr. A. K. Chakravarthy, Professor and Head, Department of Entomology and Dr. M. Thippaiah, Assistant Professor, Department of Entomology, College of Agriculture, UAS, GKVK, Bengaluru for their unforgettable encouragement throughout my academic career. But for the affection, words of encouragement, boundless love, unflagging inspiration, interest and selfless sacrifice of my parents I would not have been what I am today. I owe all my success to the special persons in my life my father

6 35 Mr. Chandrashekara, R. C., my mother Mrs. Shantha, C.S., my brother Mr. Manjunatha Swamy, R. C. I also thank my uncles Shri. Prabhuswamy, R. C, Nataraj, R. C, Basavaraj, R. C, and Mahes, R. C, and aunts Sau. Sudha, Katyayini, Sunanda and Savitra, my cousin brothers Gnanesh, Kiran, Manoj, Prajwal, Dhanush, my cousin sisters Shwetha, Aishwarya and Panchami, my Grandfather Late. Shri. Channabasavaiah, and Grandmother Sau. Mallamma without their love and blessings this study would have scarily accomplished. I owe thanks from depth of my heart to my Friends, Vijay, Kolekar, Gitesh, Waghmode, Ningot, Narayan, Anand, Mansingh, Ganesh, Subodh, Vibhav and my all Ph.D, Sr.M.Sc. and Jr. M.Sc. friends for their love, co-operation and lovely company during the M.Sc. (Agri.) degree programme. No words could justice to express my deepest sense of love and gratitude towards my friends and classmates Nikhil, Deepak, Kalpesh, Vivek, Vishal, Prashanth and Namrata for their excellent company, inspiration, moral support, boost up and healthy friendship. I owe thanks from depth of my heart to my well wishers Shabbu, Rudresh, Pallu, Kutti, Chikkesh, Madhu, Shivu, Rajeev, Arun, Thontu, Bharath, Cikada, Puneeth, and Anil, for their valuable suggestions, moral support, constant encouragement and above all being my friends. Once again, I would like to thank each and every person who helped me directly or indirectly to turn this dream come true. Place: Dapoli Date: (Dileep, R. C.)

7 36 Title of thesis Name of the student DEPARTMENT OF AGRICULTURAL ENTOMOLOGY COLLEGE OF AGRICULTURE, DAPOLI : Performance of egg parasitoid Trichogramma chilonis (Ishii under laboratory conditions : Mr. Dileep, R.C. Regd. No. : 2078 Name and Designation of : Dr.(Mrs.) K. V. Naik the Research Guide M.Sc. (Agri.), Ph.D. Assistant Professor, Department of Agril. Entomology, College of Agriculture, Dapoli. Dist. Ratnagiri (M.S.) PIN Year of award of degree : 2012 THESIS ABSTRACT The present investigations were undertaken to study the performance of egg parasitoid Trichogramma chilonis (Ishii) under laboratory conditions during the year in the Bio-control laboratory, Department of Agricultural Entomology, College of Agriculture, Dapoli (Maharashtra). The results of development of Trichogramma chilonis (Ishii) on Corcyra cephalonica (Stainton) and Spodoptera litura (Fab.) revealed that the development of T. chilonis was found superior over eggs of S. litura except in per cent parasitization on eggs of C. cepholonica. Effect of host egg age on the performance of T. chilonis revealed that egg age from 0-1 to h old were highly preferred for parasitization with maximum female recovery in egg age 0-1 to h old. Sex ratio was female biased in fresh eggs. Effect on different colour trichocards on the performance of T. chilonis revealed that for mass production of the parasitoid in the laboratory trichocard of green colour was most suitable based on results of free choice open light condition. Effect of cold storage at 15 0 C temperature for varying period on already parasitized egg cards, 4 day after parasitization revealed that parasitized trichocards

8 37 after 4 days of parasitization can effectively stored up to 30 days without much effect on adult emergence. Studies on insecticide safeties to T. chilonis realized that no insecticide was totally safe in contact toxicity study however, the ranking allotted indicated that azadirachtin ppm (0.004 %) and novaluron 10 EC ( %) were slightly harmful (Rank 2) while emamectin benzoate 5 SG ( %) was moderately harmful (Rank 3), while all remaining were totally harmful (Rank 4). Insecticide safeties to egg parasitization revealed that based on ranking, acephate 75 SP (0.1 %), imidachloprid17.5 SL (0.005 %), emamectin benzoate 5 SG ( %) and novaluron 10 EC ( %) as harmless (Score 1 = < 30 % reduction in egg parasitization over control) azadirachtin ppm (0.004 %), spinosad 45 EC ( %) and cypermethrin 25 EC ( %) as slightly harmful (Score 2 = 30-79% reduction in egg parasitization over control) while, triazophos 40 EC (0.05 %) and lambda cyhalothrin 5 EC (0.003 %) as moderately harmful (Score 3 = 80-99% reduction in egg parasitization over control). Effect of insecticides on adult emergence revealed based on the ranking given azadirachtin ppm (0.004 %) was found safe (Score 1 = < 30 % reduction in egg parasitization over control) to developing adults of Trichogramma inside the egg. Insecticides like acephate 75 SP (0.1 %), emamectin benzoate 5 SG ( %), novaluron 10 EC ( %), triazophos 40 EC (0.05 %) and cypermethrin 25 EC ( %) were slightly harmful (score 2 = 30-79% reduction in egg parasitization over control), imidachloprid17.5 SL (0.005 %) and lambda cyhalothrin 5 EC (0.003 %) as moderately harmful (Score 3 = 80-99% reduction in egg parasitization over control) and spinosad 45 EC ( %) recorded as harmful (Score 4 = > 99% reduction in egg parasitization over control).

9 38 CONTENTS CHAPTER PARTICULARS PAGE NO. I INTRODUCTION 1-4 II REVIEW OF LITERATURE 5-30 III MATERIALS AND METHODS IV RESULTS AND DISCUSSION V SUMMARY AND CONCLUSION LITERATURE CITED i-xiii APPENDIX i-ii

10 39 LIST OF TABLES TABLE NO. TITLE PAGE NO. 1. Different host egg age Different Trichocard Colours Storage period of parasitized trichocards at 15 0 C Details of insecticide treatments Development of T. chilonis on C. cephalonica and S. litura Effect of host egg age on the performance of T. chilonis Effect of Trichocard colour on the performance of T. chilonis Effect of cold storage (15 0 C) at different periods on adult emergence of T. chilonis Evaluation of various insecticides against T. chilonis adults (Contact toxicity) Insecticides safety Effect on parasitisation of pre-treated egg cards by T. chilonis Insecticides safety Effect on adult emergence of T. chilonis

11 40 LIST OF FIGURES FIG. NO TITLE BETWEEN PAGE 1 Effect on parasitization potential Effect on adult parasitoid emergence Effect on female parasitoid emergence Effect on male parasitoid longevity (Days) Effect on female parasitoid longevity (Days) Effect on total development period of parasitoid (Days) 7 Effect of host egg age on egg parasitization by Trichogramma cholonis (Ishii) 8 Effect of host egg age on female parasitoid emergence 9 Effect of Trichocard colour on egg parasitization 10 Effect of Trichocard colour on female parasitoid emergence 11 Effect of cold storage on adult emergence from parasitized trichocards 12 Contact toxicity of different insecticides to Trichogramma chilonis (Ishii) female adults 13 Insecticides safety Effect on egg parasitization

12 41 14 Insecticides safety Effect on adult emergence LIST OF PLATES PLATE NO. PARTICULERS BETWEEN PAGE 1 Mass production and maintenance of Corcyra culture 2 Corcyra moth collection using mechanical moth collection device Oviposition cage for Corcyra female Trichocards produced from nucleus culture UV chamber for egg irradiation Mass culture of Trichograma chilonis (Ishii) in glass jars with irradiated eggs Mass production of Spodoptera culture Male and female adult of Trichigrama chilonis (Ishii) adult 9 Effect of trichocard colour on parasitization No choice test 10 Effect of trichocard colour on parasitization Free choice open light test 11 Plastic container modified for testing contact toxicity to different insecticides to Trichogramma adult Insecticide testing unit Contact toxicity 40-41

13 13 Effect of Trichocard colour

14 43 CHAPTER I INTRODUCTION The indiscriminate and non selective use of insecticides in pest management has created several problems viz., development of insecticide resistance in major crop pests, pest resurgence, destruction of natural enemies besides ill effects of toxic residue in food and environment (Phokela et al., 1990, Armes et al., 1992, Lande and Sarode, 1993). In recent years, farmers are showing increased interest towards biological control of pests following the unsatisfactory control of insect pests with insecticides. Bio-agents occupy a premier position in the crop protection sector and constitute an important component of IPM on account of specificity, safety, economy with no resistance or residue problems. The elevated awareness of the impact of pesticide use on the environment and human health has resulted in efforts to reduce reliance on chemical control. A recent report by the US Congress Office of Technology Assessment indicated that biologically based technologies such as Biological Control could be more widely used to solve pressing needs of pest management. (Anon, 1995). Biological control is the regulation of pest populations using natural enemies, including predators, parasitoids, nematodes and microbial agents (Rosenheim and Jay, 1998; Bale et al., 2008). As opposed to chemical control, biological control is advantageous because it poses no threat to human health, it is environment friendly, host specific, and probability of the host developing resistance is low. In addition, development of biological control agent is less expensive compared to development of an insecticide (Bale et al., 2008). Environment friendly nature of the biological control can save a huge amount of foreign exchange which is being spent on the import of insecticides annually. Several parasitoids and predators of different crop pests have been successfully used in pest management. After the era of indiscriminate use of synthetic pesticides, innundative release of bio-control agents as a mean of pest management subsequent to their mass production on a commercial scale has become very popular. Among the several parasitoids successfully used in the pest management strategies, Trichogrammatids are one of the most important groups of bio agents with renowned interest for the suppression of lepidopterous pests all over in India. The genus Trichogramma Westwood and closely related Trichogrammatoidea Girault parasitoids attack on egg stage of the pest. More than 150

15 44 species of Trichogrammatids, are distributed throughout the world parasitizing eggs of over 200 insect species belonging to mainly Lepidoptera, Coleoptera, Neuroptera and Diptera, the majority being Lepidoptera. They are observed in diverse habitats ranging from aquatic to arboreal. In India, about 26 Trichogrammatids are recorded; of which, Trichogramma chilonis (Ishii), Trichogramma japonicum Ashmead and Trichogramma acheae Nagaraja and Nagarkatti are of significant importance. Trichogramma chilonis (Ishii) (Hymenoptera: Trichogrammatidae), a minute wasp from a group of insects of great importance to biological control, is a natural enemy of many harmful lepidopterous insect pests of crops and vegetables. The unique character of this minute parasitoid is that, it is exclusively an egg parasitoid, therefore, play important role in pest suppression programme by destroying the early stage of the pest thereby, curtailing the use of pesticides and contributing to prevent environmental pollution. Trichogrammatids are one of the important parasitoids amenable for mass production, which can be accomplished by mass culturing its factitious host, either Corcyra cephalonica (Stainton) or Sitotroga cerealella (Olivier). Due to its amenability to mass production, this group of parasitoids has the distinction of being maximum produced and released natural enemy in the world. These minute endoparasitoids of insect eggs are released in crops or forest in large numbers (up to several millions/ha) timed as per the presence of pest eggs. Trichogramma are the most widely augmented species of natural enemy, mass-produced and field released for about 70 years in biological control efforts. Worldwide over 32 million ha area under agricultural crops and forest are treated annually with different Trichogramma species in 19 countries, mostly in China and Republic of Soviet Union (Li, 1994). The economic damage caused by lepidopteran pests on field crops and on stored grain exacerbates the problem of food security and malnutrition in many developing countries (Gressel et al., 2004). Just to quote an example, among the various polyphagous pests of economic importance, the well known is, the tobacco caterpillar, Spodoptera litura (Fab.), it is next to Helicoverpa armigera (Hub.) in economic importance at national level. It is one of the important polyphagous crop pests distributed throughout south and eastern world infesting 112 species of plants belonging to 44 families, of which 40 species are known from India. It is important polyphagous pest that has about 150 hosts including major crops like soybean, cabbage, beetroot, cotton, sorghum, tomato, etc., (Rao et al., 1993). In India, S. litura has been reported as an increasingly important pest during the rainy seasons causing

16 45 heavy yield loss (Rathi and Gopalakrishan., 2005). Dhir et al.,(1992) reported S. litura is a serious but sporadic insect pest causes economic losses of crops from per cent. To tackle this problem, farmers are using enormous pesticides. The important polyphagous pest S. litura shows resistance against insecticides like cypermethrin, fenvalerate and monocrotophos. This was the first lepidopteran pest which had developed insecticide resistance in India. Therefore, it is felt necessary to study the effective and ecofriendly approach to suppress this polyphagous pest by using Trichogrammatid. In light of the above-discussed points, the present investigation has been undertaken with the following objectives. 1. Development of T. chilonis on C. cephalonica and S. litura 2. Effect of host egg age on the performance of T. chilonis 3. Effect of different coloured trichocards on the performance of T. chilonis 4. Effect of low temperature storage on parasitized trichocards 5. Relative toxicity of insecticides for T. chilonis

17 46 CHAPTER II REVIEW OF LITERATURE The review of the work done on Performance of egg parasitoid Trichogramma chilonis (Ishii) under laboratory conditions are presented in this chapter under following sub headings. 2.1 Development of T. chilonis on Corcyra cephalonica (Stainton) and Spodoptera litura (Fab.) 2.2 Effect of host egg age on the performance of T. chilonis 2.3 Effect of different coloured trichocards on the performance of T. chilonis 2.4 Effect of low temperature storage on parasitized trichocards 2.5 Relative toxicity of insecticides for T. chilonis 2.1 Development of T. chilonis on C. cephalonica and S. litura Marston and Ertle (1973) compared the effect of eggs of Trichoplusia ni (Hbn.) and Sitotroga cerealella (Olivier) for their effect on the bionomics of Trichogramma minutum Riley. They reported that the eggs of T. ni produced 2.45 times more parasitoid females than the eggs of S. cerealella when stung individually and 2.27 times more when stung in colonies for both the eggs. Parasitoid females from T. ni were more fecund than those emerged from S. cerealella eggs when both the females were attacking T. ni. But the difference was non-significant when both the females were attacking S. cerealella or when they were attacking the host eggs from which they were reared. Parasitoid females emerged from T. ni searched 1.97, 1.43 and 1.33 times more area than those emerged from S. cerealella eggs in three different trials.

18 47 Jalali et al. (1987) evaluated eggs of different lepidopteran hosts for rearing Telenomus remus Nixon (Hymenoptera: Scelionidae). The host range test indicated that S. litura, which was the preferred host, S. exigua and C. cephalonica were also parasitized with and per cent respectively and 90 per cent emergence was obtained from these three hosts. Parasitization on H. armigera, Plusia signata F. and Agrotis segetum (Schiff.) was 19.20, and 7.90 per cent, respectively and per cent emergence was obtained on these hosts. However, T. remus could not parasitize the eggs of Tribolium castaneum (Herbust.), Galleria mellonella (L.), Phthorimaea opercula Zell., Earias vittella (Fab.), Sesamia inferense Wlk. and Plutella xylostella (L.) etc. Somchaudhury and Dutt (1988) studied the effect of host on biology of Trichogramma perkinsi Gir. and Trichogramma australicum Gir. utilizing eggs of Helicoverpa armigera (Hub.), Chilo partellus (Swinhoe), Achaea janata (L.), Exelastis atomosa Wals., Euproctis fraterna Walker, S. litura and Spilosoma obliqua Walker. The females emerging from H. armigera lived significantly longer than those from E. fraterna, S. litura and S. obliqua. The duration of life cycle was maximum in eggs of S. obliqua and minimum in eggs of A. janata. Sex ratio was adversely affected in case of T. perkinsi when reared on eggs of S. litura and in case of T. australicum when reared on E. fraterna. Effect of host egg on the biology of Trichogramma exuguum Pinto studied by Ram and Irulandi (1989) revealed that fecundity, progeny production and adult longevity was the highest in the females reared in the eggs of Euproctis lunata Walk., a larger host egg than the other two viz., C. cephalonica and C. partellus. They also revealed that apart from the size, the quality of nutrition from host egg also influence the fecundity and progeny production. They

19 48 further noticed that the adult emergence was also significantly influenced by the egg size. However, the sex ratio of the progeny of the parasitoid was non-significant. Selvaraj and Sundarababu (1995) studied the performance of various hosts on the biology of T. chilonis in laboratory. They reported that, T. chilonis showed greatest preference for the eggs of P. operculella as evidenced by the highest rate of parasitization (76.20 %), adult emergence (96.33 %), proportion of females (55.31 %) and longevity (6.4days). Therefore, P. operculella was considered more suitable for rearing the parasitoid than the other hosts viz., C. cephalonica, H. armigera and S. litura Effect of four different host eggs viz., H. armigera, Amsacta moorei Butler, E. vittella and C. cephalonica on some biological and morphological characteristics of T. chilonis was studied by Rathi and Ram (2000). The results revealed that the size of T. chilonis male and females was bigger when developed on bigger sized eggs. Similarly, the number of parasitoids emerged per eggs was more from the larger host eggs (A. moorei) as compared to the smaller ones (C. cephalonica). The adult emergence of T. chilonis was higher (93.30 %) when reared on C. cephalonica and lowest (83.30 %) on the eggs of A. moorei. The proportion of female progeny was more (74.90 %) from A. moorei eggs and lowest (68.70 %) on C. cephalonica. Similarly they found that the development period was longer (11.2 days) on H. armigera eggs and shorter (9.3 days) on eggs of C. cephalonica. They further affirmed that the female parasitoids emerging from the eggs of E. vittella were more fecund (107.5 host eggs parasitized) as compared to those from A. moorei (56.1 eggs parasitized). Further adult longevity revealed that male and female parasitoids survived longer (5.2 and 9.4 days) when reared on the eggs of E. vittella. From overall results they concluded

20 49 that the E. vittella might be used as natural host for Trichogramma rearing after every few generations to maintain the vigor and quality of the parasitoid. Hoffmann et al. (2001) studied the performance of the parasitoid, Trichogramma ostriniae (Pang and Chen)on four factitious pest s eggs viz., Ostrina nubilalis Hubner, S. cerelella, T. ni and irradiated eggs of E. kuehniella. They noticed that E. kuehniella was a poor host, T. ni and O. nubilalis were good hosts and S. cerealella was intermediate. Sheeba et al. (2002) studied on Effect of host and food on biological parameters of Trichogramma japonicum Ashmead (Hymenoptera: Trichogrammatidae) native to the Andaman islands. They revealed that honey fed females of T. japonicum on an average survived for 51.6 hrs. and produced 32 offspring; unfed females survived for 2.4 hrs and produced 19 offspring. There was reduction in parasitisation, longevity and fecundity when parasitoid was not fed on honey in comparison to honey fed parasitoid. Honey fed individuals had 1.69 times more egg laying potential than unfed ones. The survival percentage of the females subjected to adequate food source was upto 30 per cent on the third day whereas only 10 per cent on the first day itself and 0 per cent on subsequent days without food source. Tiwari and Khan (2003) revealed that T. chilonis preferred freshly laid eggs of Corcyra than S. obliqua and parasitized to the tune of 91.97and per cent, respectively. Spitzen and Huis (2005) observed that Uscana lariophaga Steffan reared on small eggs of Callasobruchus maculates (Fab.), developed slower, were smaller and produced fewer eggs compared to parasitoid reared in large hosts. Fecundity of the parasitoid was

21 50 heavily influenced by the host egg size. Similarly, they reported that wasp allocated marginally more female offspring to larger hosts. Nathan et al. (2006) reported greater percentage of T. chilonis emergence from eggs produced by C. cephalonica fed on Eleusine coracana Gaertn than from eggs produced by C. cephalonica fed on Sorghum bicolor (L.), Oryza sativa L. or Triticum aestivum L. Significantly greater percentage of T. chilonis survived 24 h after emergence from eggs produced by C. cephalonica those were fed on E. coracana than from eggs of adults fed on S. bicolor as larvae. Nabil and El-Wakeil (2007) evaluated the efficiency of Trichogramma evanescens West. reared on different factitious hosts to control H. armigera and he reported that rates of parasitism on H. armigera eggs, emergence rates of parasitoids and their longevity were the highest for wasps reared on H. armigera. Wasps reared on S. cerealella gave comparable rates. However, wasps from E. kuehniella gave the lower rates and G. mellonella gave the lowest ones. Parasitized eggs of H. armigera and S. cerealella produced more parasitoid females than eggs of E. kuehniella and G. mellonella. Budhwanth et al. (2008) studied efficacy of T. chilonis against lepidopteran pests and age of host eggs. They reported that out of 30 eggs exposed to T. chilonis significantly maximum number of eggs of H. armigera were parasitized of eggs followed by S. litura (18.78 eggs) and significantly lower in Papilio demoleus (Linn.) (7.11 eggs). Less parasitized eggs were recorded in P. demoleus due to its large size of egg (0.82 mm 2 ) as compared to H. armigera (0.23 mm 2 ) and S. litura (0.30 mm 2 ). Similarly, per cent parasitization was also found maximum in case

22 51 of H. armigera (65.10 %) followed by S. litura (59.20 %) and significantly lower in P. demoleus (22.90 %). Nadeem et al, (2009) reported that biological parameters of parasitoids, T. chilonis such as parasitism (95.60 %), developmental period (7.3 days), emergence (98.00 %) and adult longevity (9.0 days) were very favourable at 28 0 C followed by at 25 0 C (92.80%, 8.3 days, %, and 10.0 days, respectively). Roopa et al. (2009) studied biological attributes of arrhenotokous (sexual) and thelytokous (asexual) forms of Trichogramma species. They reported that per cent parasitization in arrhenotokous species varied from , compared to %, in thelytokous species. Mean longevity of arrhenotokous forms was 6.64 days compared to 9.45 days in thelytokous species. Developmental period was shorter in thelytokous forms, whereas mean fecundity was higher in arrhenotokous species. 2.2 Effect of host egg age on the performance of T. chilonis Huis et al. (1991) conducted an experiment on selection of hosts of different ages in case of egg parasitoid U. lariophaga and its hosts viz., C. maculates and Bruchidius atroliniatus Booker. The results revealed that at 30 0 C, C. macualtus eggs of 0 to 2 day old were readily accepted for parasitisation, but in a choice situation freshly laid eggs were preferred. Of the older eggs, only those of 3 day old were preferred but significantly less than those of 0 to 2 day old. Similarly at 30 0 C, B. atrolineatus eggs older than 24 hours were significantly less parasitized than younger eggs. It was also revealed that when eggs were older than one day, the time taken for penetrating the eggs increased with host age.

23 52 Miura and Kobayashi (1998) studied the effects of host egg age on the parasitism by T. chilonis (Hymenoptera: Trichogrammatidae), an egg parasitoid of the Diamond Back Moth (DBM). They reported that females oviposited on host eggs of all ages tested. However, the percentage of oviposition was highest on the 1 day old host eggs, and decreased as the host egg became younger or older than the 1 day old host. Parasitized host eggs did not hatch. The emergence rate was high on the 1 day old eggs but did not differ among other host ages, except the 3 day old host. T. chilonis progeny failed to complete development on the 3 day old DBM eggs. Takada et al. (2000) studied biological characteristics: growth and development of the egg parasitoid Trichogramma dendrolimi Matsumura on the cabbage armyworm Mamestra brassicae (L.). They reported that T. dendrolimi was able to parasitize M. brassicae eggs of all ages. However, successful parasitism significantly decreased on 4 day and 4.2 day old host eggs. The female ratio of the progeny decreased with the aging of the female wasps, but neither the sequential change of the host age nor variation of the host egg mass size affected the female ratio of the progeny which was about 80 per cent. Singh et al. (2001) studied the effect of host egg age on two egg parasitoids viz., T. brasiliensis and T. exiguum with 0 to 73 h old eggs of C. cephalonica. They observed that per cent parasitisation in both the parasitoids was the maximum in case of fresh eggs which declined as the egg age increased. Similarly, adult emergence in both the parasitoid species also decreased as the age of host egg increased. In case of female longevity, it was noticed that there was no effect on the female longevity of T. brasiliensis upto 66 to 67 h, however, lowest female longevity of 2 days was recorded in 72 to 73 h old eggs. Further the longevity of T. exiguum female decreased as

24 53 the age of the egg increased. Data on fecundity of T. exiguum revealed that eggs of age 54 to 55 h old had no effect on the fecundity. Highest fecundity of 98.7 was recorded from eggs of 6 to 7 h old age and no egg laying was observed in the females emerged from eggs of 72 to 73 h old. In T. brasiliensis fecundity same pattern was noticed with lowest fecundity of 10.3 from females emerged from 72 to 73 h old eggs. Tiwari and Khan (2003) studied the preference of T. chilonis for different hours-old eggs of S. obliqua. Results emanating from the studies revealed that freshly laid eggs (0 h old) were most preferred with mean parasitization of %, while 24 and 48 h old eggs had insignificant differences regarding parasitization levels. Influence of host quality on rearing the native egg parasitoid, T. japonicum was studied by Singh and Prasad (2003). They evaluated ultra-violet irradiated various age group (6 to 78 h old) eggs of C. cephalonica. Per cent parasitization, adult emergence, sex ratio, longevity and fecundity were evaluated. The results revealed that older eggs were less suitable for parasitization by T. japonicum; further host age was positively correlated with the number of female offspring of T. japonicum. Longevity and fecundity declined with the increase in host age. Based on the results, they concluded that use of younger, fresh eggs (6 to 18 h old) irradiated for 50 to 55 minutes was most efficient for increasing the reproductive efficiency of T. japonicum. Reznik and Vaghina (2007) studied effect of experience on response of Trichogramma buesi Voeg. and Trichogramma principium Sug. females to different ages of host eggs. They reported that the pattern of T. principium and T. buesi female distribution by number of mature ovarial eggs was strongly dependent on age of the offered

25 54 host (S. cerealella) eggs. The percentage of parasitizing females roughly estimated from these distributions was times higher for females offered fresh host eggs than for those offered "old" eggs, that had developed 6 days at a temperature of 20 0 C. In both species, females more often oviposited in old (non-preferred) eggs when they had been previously experienced with fresh (preferred) eggs. Zahid et al. (2007) conducted experiment on the effects of parasitoid and host egg age on parasitism by T. chilonis. They reported that T. chilonis laid significantly the highest average numbers of eggs at and per cent during the first 8 and 24 h of its age, respectively. Later, the parasitization decreased to 35 per cent by the end of day 4. Thus, for the highest yield of parasitoid production, it is important to use younger Trichogramma for parasitization. Budhwanth et al. (2008) studied efficacy of T. chilonis against lepidopteran pests and age of host eggs. They reported that maximum parasitization (60.30 %) was noticed in fresh eggs of 24 hrs old followed by 48 hrs old eggs (49.50 %) while, 72 hrs old eggs were less preferred (37.30 %) by T. chilonis for parasitization. However, the different lepidopteran pests as well as age of host eggs did not show significant differences pertaining to per cent adult emergence. Fresh eggs of 24 hrs could produce more female progeny of T. chilonis (52.90 %) compared to old age of 72 hrs (48.70 %). Mehendale (2009) studied effect of host egg age on the performance of T. chilonis. He reported that egg age from 0 to 1 to 24 to 25 h old were highly preferred for parasitization with maximum female recovery in egg age 0 to 1 to 18 to 19 h old.

26 Effect of different coloured trichocard on performance of T. chilonis No much work is available on this aspect. However, Romies et al. (1998) studied the response of Trichogramma egg parasitoid to colour sticky traps in a field study in two seasons. Colours tested were white, green, blue, yellow and red in the first season and white, green, yellow and black in the second season. The proportion of both female and male parasitoids caught on the sticky traps was significantly different among colors, indicating that the parasitoids actively move between plants and are not solely carried along passively by wind. White colour was most preferred by female parasitoids, followed by clear and green traps. Yellow was preferred over black but was less attractive than green. Visual cues may be used by Trichogramma spp. during the habitat location process. The colour preference of male Trichogramma spp. differed significantly from female with yellow and green being more attractive than white. For all colors, more Trichogramma spp females were caught on the sticky traps (>85% of all wasps caught), indicating a lower activity level and/or shorter lifespan of males. Baitha and Sinha (2002) studied the effect of trichocard colour on some biological attributes of T. chilonis (Sugarcane strain from Lucknow) in the laboratory. Colours tested were white, orange, yellow, red and green. Longevity of female T. chilonis varied from 5 to 8 days on different colours of trichocard. Green colour supported maximum and significantly higher longevity (8 days) of female individuals than other colours. Significantly higher fecundity (number of eggs) was recorded on white trichocard (110) followed by green (86.80). Green trichocard supported maximum and significantly higher emergence of adult (76.29 %) and females (76.46 %) than on other colours. Rate of oviposition in the first

27 56 3 days was the maximum on white (90.4) followed by green trichocard (76.6). Oviposition in rest of the period (4-8 days) was the maximum on white trichocard (19.6) followed by yellow (10.4) or green (10.2). Comparing white and green colours, the percentage of oviposition was highest (88.64 %) on green trichocard in the first 3 days than on white (84.62 %). Thus they concluded that for mass multiplication of T. chilonis in laboratory, trichocard of white or green colours were most suitable. Bhattacharya et al. (2003) worked on the effect of the colour of cards used to fix C. cephalonica eggs on the behaviour and biological attributes of the parasitoid T. chilonis. Under multiple-choice conditions, the per cent parasitization varied significantly on differently coloured egg cards. The maximum parasitization was observed on mint-green coloured egg cards (64.7 %), followed by jade green (56.75 %), golden yellow (54.25 %), deep orange (52.25 %), bus green (49.25 %), canary yellow (46.25 %) and white (40.50 %). The lowest parasitization was observed on chassis gray (11.50 %), followed by dawn (14.50 %) and wild lilac (15.75 %) coloured cards. The maximum emergence was on mint green cards (88.06 %), followed by golden yellow (88.04 %) and white (87.76 %). The lowest emergence was observed in wild purple egg cards (86.05 %). The highest female emergence was observed on white cards (72.63 %), while the lowest was on sky blue cards (71.09 %). In single choice conditions, the maximum fecundity was on mint green cards (87.50 %), followed by deep orange (87.33 %) and golden yellow (87.17 %). The maximum adult emergence was on white cards (87.00 %). Results of multiple and single choice experiments indicated that the insect preference was higher for mint green, jade green, golden yellow deep orange and white colour trichocards.

28 57 Lobdell et al. (2005) studied the host colour preferences and short range searching behavior of the egg parasitoid T. ostriniae. They reported that the colour of lepidopteran eggs often varies by species or egg condition, and parasitoids that attack lepidopteran eggs could, therefore, potentially use colour to obtain information about host identity or quality. They examined the wasps' hostselection behavior in a petri dish arena using white, yellow, green, and black clay beads as egg models presented against a green background (to mimic leaf colour). In no-choice tests, bead colour had a significant effect on the proportion of tested wasps that accepted a bead for further examination, on the time it took wasps to find and begin examining a bead, and on the time that wasps spent examining the beads. However, bead colour had only a marginally significant effect on the proportion of wasps attempting to drill into a bead with their ovipositors, and no significant effect on the amount of time they spent drilling. The wasps also showed significant colour preferences when given a choice between two adjacent beads of different colours. The results of the no-choice and choice trials taken together indicated a colour preference ranking of yellow > white > green > black. The wasps' higher preference for the yellow and white egg models generally corresponded to the white or yellowish-white egg colour target host of T. ostriniae, the European corn borer moth, O. nubilalis (Lepidoptera: Crambidae). The wasps strong rejection of black egg models was likely to be an adaptive response that reflected the fact that eggs that were wholly or partially black were often unsuitable for parasitization due to advanced caterpillar development, damage to the egg, or previous parasitization. Vishla et al. (2007) studied the effect of card colour on the parasitization and emergence of Trichogrammatids. They reported

29 58 that the percent parasitism differed significantly on eggs cards of different colours. T. chilonis showed maximum parasitism on yellow coloured cards (81.37 %), T. japonicum showed maximum parasitization on signle red (52.12 %) and T. brasiliensis on satin blue (82.00 %). whereas T. pretiosum showed maximum parasitism like T. japonium on single red coloured card (49.75 %). However, percent adult emergence of all Trichogrammatids on different coloured egg cards showed non-significant from each other. Mehendale (2009) worked on effect of different colour trichocards on the performance of T. chilonis. He reported that in free choice open light conditions, trichocard colours viz., yellow, green or blue followed by white supported higher parasitization. However, maximum per cent female parasitoid emergence was recorded only in green colour from all three situations and concluded that paper of green colour would be most suitable for preparing trichocards. 3.4 Effect of low temperature storage on parasitized trichocards Venkatraman and Govil (1951) studied the effect of low temperature (refrigeration) on the viability of T. evanescens minutum. The results revealed that the percentage of survivals decreased as the period of refrigeration increased. Up to about 15 days of refrigeration there was no much effect on viability. According to reports of Singh and Jalali (1994), T. minutum can be stored in pre-pupal/pupal stage at 12 0 C and 85 per cent relative humidity up to 25 days; T. brasiliense at 10 0 C up to 50 days and Tr. eldanae at 5 0 C up to 88 days in the refrigerator before field release. Singh et al. (1997) reported that seven days old eggs parasitized by T. chilonis could be stored for 20 days in the

30 59 refrigerator without affecting the adult emergence, parasitization efficiency and sex ratio. According to Khosa and Brar (2000), the parasitoid T. chilonis could be stored in the refrigerator and successfully utilized for 23 days without adversely affecting their emergence and parasitisation efficiency. Tezze and Botto (2001) evaluated the possibility of storing Trichogramma nerudai Pintureau and Gerding at low temperature and the effects of such storage on the biological quality of the parasitoid and its progeny were studied. Pupae of T. nerudai were stored 25, 50, 75, 100, 125 and 150 days at 4 ± 1 C and relative humidity 75 ± 5 per cent in a refrigerator, and full darkness. T. nerudai pupae were found tolerant to cold storage. Important components of the biological quality in Trichogramma viz., the number of emerged adults, proportion of deformed adults, the flight and the mobility capacity were not seriously affected until 50 days of cold storage. However, the biological quality of the parasitoids was significantly affected by cold storage from 75 days onwards. From the results, they concluded that cold storage was useful to store T. nerudai pupae up to 50 days. A laboratory experiment for determination of storage conditions conducive for holding Trichogrammatoidea bactrae Nagaraja was conducted by Gupta and Bhardwaj (2002). The results revealed that from parasitized eggs of C. cephalonica, stored after 3-7 days of parasitisation for 5 days at 10 0 C, above 50 per cent adult emergence occurred and eggs could not be stored for more than 7 days. However, at 15 0 C, storage could be extended up to 18 days but with low emergence (17.2 %). In another experiment, exposure to 10 and 15 0 C for 5, 10 and 15 days after 3, 5 and 7 days of parasitisation provided satisfactory adult emergence from eggs

31 60 stored at 15 0 C for 5 days after 5 and 7 days with parasitisation of and per cent, respectively and for 10 days after 5 days of parasitisation with per cent parasitization. SuChiung and ShengChih (2004) studied the adult emergence rate after cold storage at 4 ± 2 0 C for 3 to 42 days in case of T. chilonis and revealed that various ages of T. chilonis after 6 cold storage days, the adult emergence rates were all over per cent, the 4-day old T. chilonis after 3 to 42 days of cold storage, the adult emergence rates of all treatments were higher than per cent. Thus they found that for cold storage, 4-day old parasitized eggs were suitable. Kumar et al, (2005) studied on effect of low temperature storage on the efficiency of three species of trichogrammatids. They reported that T. chilonis and T. pretiosum can be stored for 20 days whereas T. brasiliense for 10 days without adversely affecting their emergence and parasitisation efficiency taking per cent emergence and per cent parasitism as standard. The emergence in T. brasiliense reached zero per cent after 34 days of storage, whereas per cent was recorded in T. chilonis and per cent in T. pretiosum after 60 days of storage. T. chilonis showed the highest emergence and parasitization efficiency followed by T. pretiosum and T. brasiliense. Vishla et al. (2008) worked on the effect of storage period on the emergence and parasitization efficacy of Trichogrammatids. They reported that the parasitized eggs of C. cephalonica stored at 10 C showed decreasing trend of parasitoid emergence with the increase in storage period. It reached per cent from per cent after 30 days of storage. The parasitization efficacy of adults Trichogrammatids also decreased with the increase in storage period, which reached per cent from per cent

32 61 after 10 days. Wherein, Trichocards can be stored for 10 days at 10 C without significant reduction in the emergence and parasitization efficacy of Trichogrammatids female. Mehendale (2009) worked on effect of cold storage under refrigerated conditions at 15 0 C temperature for varying periods on 4 days after parasitization of trichocards. He reported that per cent adult emergence was quite high at 5 days storage (89.0 %) followed by 10 days (86.0 %), 15 days (79.31 %), 20 days (68.47 %), 25 days (63.97 %) and 30 days (63.80 %). Thereafter, the adult emergence declined drastically. He further concluded that parasitized trichocards can be stored effectively up to 30 days at 15 0 C. Nadeem et al. (2010) worked on optimization of short and long term storage duration for T. chilonis at low temperatures. They revealed that highest (96.60 %) T. chilonis emergence recorded at 10 0 C after 5 days storage and was similar to control (97.40 %). However, emergence was reduced up to per cent at 10 0 C after 90 days storage. The parasitism was highest (97.40 %) at 10 0 C after being kept for 5 days storage and was decreased to per cent at 10 0 C when stored for 90 days at same temperature. Adult longevity was decreased from 6.3 to 3.0 days when stored at 10 0 C from 5 to 90 days, respectively. Pathak et al. (2010) studied the suitability of temperature for the storage of T. chilonis. They reported that among the treatment combinations, best treatment proved to be storage of cards at 15 C for 30 days after 5 days of parasitization, when highest parasitoid emergence was observed (72.4%). Ahmad et al. (2011) studied the parasitism potential and low temperature storage response of Trichogramma kashmirica Nagaraja, Ahmad and Gupta. They revealed that the per cent

33 62 emergence and females declined from to and to 86.80, respectively, after storage for 45 days at 6 C. Adult emergence and longevity of females were negatively correlated, whereas developmental period was positively correlated with storage period. 2.5 Relative toxicity of insecticides for T. chilonis Sithanantham and Paul (1980) assessed the persistence toxicity of six insecticides viz., chlorpyriphos, endosulfan, endrin, endrin+ thiometon, leptophos and phosalone to the developing and adult stages of T. australicum. They reported that sugarcane foliage sprayed with these insecticides retained medium lethal levels up to 9, 1, 7, 6, 7 and 7 days, respectively. The insecticides also affected emergence of the parasite. Further, endosulfan was least toxic to all stages of the parasite, while chlorpyriphos was highly toxic to the developing stages. Verma et al. (1988) evaluated the effect of 15 insecticides used in cotton on the susceptibility and emergence of T. achaeae under laboratory conditions. All the insecticides except fenvalerate gave cent per cent mortality. Maximum parasitization was observed when the unparasitized eggs of C. cephalonica were treated with permethrin, oxydemeton methyl and fenvalerate. When the insecticides were tested on parasitized eggs, it was found that fenvalerate, permethrin, oxydemeton methyl, DDT, dimethoate, deltamethrin and phosphamidon were comparatively safe to this parasitoid. Emergence of the parasitoid was significantly affected in 1 and 2 days old parasitized eggs. Emergence in all the insecticidal treatments was significantly reduced. Paul and Agarwal (1989) studied the persistent toxicity of some insecticides in cotton against T. brasiliensis where they

34 63 revealed that carbaryl showed the highest persistence toxicity to the parasitoid followed by triazophos. Among the synthetic pyrethroids, fenvalerate showed higher persistence. Among all the insecticides tested, cypermethrin recorded the lowest persistence followed by deltamethrin, flucythrinate and fenpropathrin indicating their low toxicity to the parasitoid. Rajendran and Hanifa (1997) revealed that two insecticides viz., endosulfan and 2000 ppm were found to reduce the emergence of parasitoid from the egg card to the extent of 43.2 and 34.1 per cent during and 50.3 and 42.1 per cent during seasons. Correspondingly in the untreated plots emergence was up to 78.2 and 85.8 per cent in these two seasons. Oznipar and Kornosor (1998) studied the effect of four different insecticides recommended for the control of corn pests on the adult emergence of T. evanescens (Hymenoptera, Trichogrammatidae) from the parasitized host eggs of O. nubilalis (Lepidoptera, Pyralidae) at field conditions. In the plots, which were sprayed with monocrotophos, cyfluthrin, thiodicarb and cypermethrin, the adult emergence from the parasitized host eggs was 58.71, 49.98, and per cent, respectively. The adverse effect of all insecticides was considerably higher than the untreated plot. According to them, monocrotophos was considered to be relatively safe to T. evanescens. The effect of the remaining insecticides was similar. Kumar and Santharam (1999) evaluated the effect of imidacloprid in laboratory against T. chilonis. They revealed no significant adverse effect on adult emergence and per cent parasitisation of T. chilonis.

35 64 Effect of insecticides on emergence, adult survival and fitness parameters of T. exiguum was studied by Charles et al. (2000). Insecticides tested were lambda cyhalothrin, cypermethrin, thiodicarb, profenophos, spinosad, methoxyfenozide and tebufenozide. All insecticides, with the exception of methoxyfenozide and tebufenozide, adversely affected emergence of Trichogramma from eggs of Helicoverpa zea (Boddie) when exposed at different preimaginal stages of development (larval, prepupal or pupal). Based on LC50 values, spinosad and prophenophos were the most toxic compounds to T. exiguum adult females, followed by lambda cyhalothrin, cypermethrin, and thiodicarb. Insecticides which were field-weathered for 4 to 6 days on cotton leaves showed no activity against female T. exiguum adults. Geethalakshmi and Chandrasekaran (2000) studied the safety of chlorpyriphos and monocrotophos at 100, 50, 25, 10, 5 and 1-ppm concentrations for T. chilonis under laboratory conditions. They revealed that chlorpyriphos was more toxic than monocrotophos which showed reduced per cent parasitization and no adult emergence. Neem based and Bt formulations of biopesticides alonwith endosulfan were evaluated by Thakur and Pawar (2000) for their relative toxicity to newly emerged adults of T. chilonis. Observations revealed that neem-based pesticides and bio-pesticides were harmless while endosulfan was slightly toxic. Prem et al. (2001) studied the effect of four commercially used insecticides on tomato crop on parasitization and adult toxicity in laboratory. Acephate (0.05 %) proved safer to the parasitized host as well as to the adult parasitoid; however the adult emergence was significantly lower from treated eggs than the untreated eggs.

36 65 Tezze and Botto (2002) investigated the side effects of insecticides viz., betacyfluthrin, triflumuron, B. thuringiensis var. Kurstaki and an aqueous extract of Melia azedarach L. leaves against T. nerudai. The dosages were the maximum recommended for field use. Emergence of adults for all the treatments did not show significant differences from the control group (ranging between and %) except for the beta-cyfluthrin treatment (49.00 %). Parasitization was studied as a sub lethal effect for triflumuron, B. thuringiensis var. Kurstaki and the aqueous extract of M. azedarach leaves and no significant differences were found comparing with the control group. Tiwari and Khan (2002) studied the effect of fenobucarb and chlorpyriphos methyl at different concentrations against T. chilonis. Chlorpyriphos methyl (0.30%) and fenobucarb (0.50%) recorded 44.0 and per cent egg parasitization, respectively. While higher per cent parasitisation of and in same order of insecticides was recorded at 0.05 per cent concentration, respectively. Control recorded per cent parasitization. Carvalho et al. (2003) studied the side effect of lufenuron (0.4g a.i. /L), trifiumuron (0.15g a.i. /L), imidacloprid (0.28g a.i. /L), cyromazine (0.11g a.i. /L), methoxifenozide (0.12g a.i. /L), pirimicarb (0.25g a.i. /L) and abamectin (0.18g a.i. /L) on different developmental stages of the egg parasitoid T. pretiosum. Abamectin was the only insecticide to affect parasitoid emergence and sex ratio, regardless of the developmental stage and parasitoid generation exposed. Abamectin, lufenuron and pirimicarb also decreased the lifetime of F1 females exposed during the egg-larva stage. All the products significantly reduced the capacity of parasitization when females were treated in pupal stage.

37 66 Tiwari and Khan (2004) studied the effect of endosulfan at different concentrations on per cent parasitisation by three species of Trichogramma viz., T. chilonis, T. japonicum and Trichogramma poliae Nagaraja and revealed that maximum parasitisation was recorded at 0.25 per cent concentration of endosulfan in all the three species. At 1 per cent concentration, T. japonicum recorded the lowest of 35 per cent parasitisation among the three species. Baladandi et al. (2005) observed the side effects of maximum recommended doses (for field use in sugarcane) of six insecticides viz., endosulfan 35 EC, dimethoate 30 EC, chlorpyriphos 20 EC, monocrotophos 36 WSC, profenophos 50 EC, oxy-demeton methyl 25 EC on T. chilonis, T. japonicum and Ooencyrtus papilionis Ashmead. Based on reduction in parasitization, the insecticides were categorized into four categories, 1: harmless (< 30%), 2: slightly harmless (30 to 79%), 3: moderately harmful (80 to 99%) and 4: harmful (> 99%). Results revealed that the insecticides differed in their initial toxicity. Dimethoate (0.03%), oxy- demeton methyl (0.03%) and monocrotophos (0.05%) were slightly harmful to T. chilonis but moderately harmful to T. japonicum and O. papilionis. However, endosulfan (0.07%) was also moderately harmful to both Trichogramma species and O. papilionis. Chlorpyriphos (0.01%) and profenophos (0.006%) were harmful in initial toxicity tests on all the three adult parasitoids. Singh and Kaur (2005) tested the effects of some insecticides on one-day-old Trichogramma adults under laboratory conditions. Adults reared, on the alternate host C. cephalonica, were exposed to surface treated vials. Only endosulfan, dimethoate and monocrotophos were found less toxic to T. brasiliensis on the recommended field doses, whereas malathion and chlorpyriphos

38 67 showed 100 per cent mortality within 15 minutes of exposure. Biopesticide B. thuringiensis did not affect the parasitoids. Virk and Brar (2005) evaluated T. chilonis in combination with different insecticide sprays in cotton crop. They found that in field, highest mean per cent bollworm eggs were parasitized in control (6.78%) followed by monocrotophos (5.24%), ethion (5.06%), acephate (4.91%), quinalphos (4.88%), endosulfan (4.86%) and deltamethrin (4.72%). This indicated that all the insecticides could be safely integrated with the parasitoid. Butter and Dhalival (2006) studied on effect of azadirectin based formulations (Econeem) on the natural enemies of H. armigera. They reported that mortality of adults of T. chilonis was 1.33, 4.00 and per cent in , and ppm of Econeem, respectively. The adult emergence of T. chilonis got affected when parasitized host eggs were treated with Econeem at (84.33%), (82.88%) and ppm (77.77%) concentration. Effect of deltamethrin on the reproduction of Trichogramma cordubensis Vargas and Cabello, a thelytokous egg parasitoid was studied by Garcia et al. (2006). The results revealed that offspring emergence was significantly influenced by the insecticide treatments evaluated on their progenitors, decreasing significantly at 48 and 72 hours for the highest tested concentration of deltamethrin (23.6 mg a.i. /L). Despite that, deltamethrin had no adverse effects on the reproduction of treated wasps, particularly when was applied at the concentration recommended by the manufacturer (12.5 mg a.i. /L). Samantha et al. (2006) evaluated the residues of different insecticides in/on brinjal and their effect on T. chilonis and T. japonicum. They showed that alpha cypermethrin was found to be

39 68 safest towards both the species. Considering the retention period of the toxicant, they recommended to release both the parasitoids in the cropping ecosystem after 3 to 5 days of alpha cypermethrin, 4 to 6 days of methomyl and 6 to 7 days of quinalphos spray, respectively depending upon the treatment doses. Basappa (2007) studied on toxicity of biopesticides and synthetic insecticides to egg parasitoid, T. chilonis under laboratory conditions. Neem seed kernel extract (NSKE 5%), custard apple seed extract (CASE 5%), pongamia seed extract (PSE 5%), neem oil (2%) and pongamia oil (2%) were compared with commercial neem formulation (0.2%) for their toxic effect Synthetic insecticides like imidacloprid (0.01%), acetamiprid (0.002%), thiomethoxam (0.02%), profenophos (0.05%), and carbosulfan (0.14%) were compared with monocrotophos (0.05%) for their toxicity to T. chilonis. Most of the botanicals were found safe to T. chilonis. Among six insecticides tested, carbosulfan was found to be highly toxic to T. chilonis, followed by acetamiprid. Imidacloprid was relatively less toxic to T. chilonis. Microbial biopesticide, Bacillus thuringiensis (Berlinear) was also found safe to T. chilonis. Babasaheb et al. (2009) worked on effect of some newer insecticides along with microbials and a botanical on parasitization efficacy and mortality of T. chilonis reared on UV irradiated and unirradiated H. armigera eggs. They revealed that x 109 POBs/ml and B. 2ml/L. were safer to the parasitoid whereas neem oil 1.0% and lamda-cyhalothrin % reduced the parasitization. The lambda-cyhalothrin % adversely affected the emergence of parasitoid. The neem oil did not adversely affect the development and emergence of parasitoid although it had reduced the parasitization. Among the insecticides tested, a synthetic pyrethroid, lambda-cyhalothrin % was relatively

40 69 less persistent over the other conventional insecticides whereas imidacloprid % and spinosad 0.01 % had high residual toxicity to T. chilonis adults. Kaur et al. (2009) screened some insecticides against egg parasitoid T. brasiliensis. They found that seven insecticides were extremely toxic to adults, while four slightly toxic and two chemicals were harmless. At pupal stage abamectin and imidaclorprid were slightly toxic with > 50 per cent emergence but extremely toxic to emerged adults after 24 hours i.e. < 10 per cent survival. Endosulfan, monocrotophos and cypermethrin were slightly for adult and pupa with maximum survival in cypermethrin. Mehendale (2009) assessed safer insecticides against T. Chilonis. He reported that no insecticide was totally safe in contact toxicity study, however, ranking allotted indicated that novaluron 10 EC ( %) was slightly harmful while emmamectin benzoate 5 SG (0.001 %) and endosulfan 35 EC (0.075 %) were moderately harmful while remaining all were totally harmful. Insecticides safeties to egg parasitisation revealed that based on ranking, imidacloprid 17.8 SL (0.005 %), acetamiprid 20 SP (0.004 %), acephate 75 SP (0.1 %), emamectin benzoate 5 SG (0.001 %) and novaluron 10 EC ( %) as harmless, triazophos 40 EC (0.05 %) and cypermethrin 25 EC (0.005 %) as moderately harmful, while, fenobucarb 50 EC (0.1 %) and endosulfan 35 EC (0.075 %), the only harmful. Effect on insecticides on adult emergence revealed that, no insecticide was safe found safe to developing adult trichogramma inside the egg. Insecticides like trizophos 40 EC (0.05 %), acetamiprid 20 SP (0.004 %), acephate 75 SP (0.1 %), emamectin benzoate 5 SG (0.001 %), novaluron 10 EC ( %) and endosulfan 35 EC (0.075 %) were slightly harmful,

41 70 imidacloprid 17.8 SL (0.005 %) as moderately toxic and fenobucarb 50 EC (0.1 %) as the most toxic one. Preetha et al. (2009) studied on toxicity of selected insecticides to T.chilonis and their safety in rice eco system. They reported that all the insecticides tested showed different degrees of toxicity to the parasitoid. Thiamethoxam showed the highest toxicity to T. chilonis with an LC50 of mg a.i. /L, followed by imidacloprid ( mg a.i. /L). The LC50 values of acephate and endosulfan were and mg a.i. /L, exhibiting low toxicity when compared with other insecticides tested. Thiamethoxam was found to be 3,195, 1,395 and 1,322 times more toxic than acephate, chlorantraniliprole and endosulfan, respectively, as revealed by the LC50 values to T. chilonis. Based on risk quotient, which is the ratio between the field-recommended doses and the LC50 of the beneficial, only chlorantraniliprole was found to be harmless to T. chilonis. The insecticides thiamethoxam, imidacloprid, Virtako and ethofenprox were found to be dangerous to the parasitoid. Almeida et al. (2010) studied the effect of azadirachtin on the control of Anticarsia gemmatalis (Hub.) and its impact on T. pretiosum. They revealed that azadirachtin did not affect negatively the parasitism, emergence or sex ratio of the progeny. This indicates that the product can be used with mass release of T. pretiosum to control A. gemmatalis. Shoeb (2010) studied the effect of five insecticides, Profect (w.p.), CAPL- 2, Lambda-cyhalothrin, Spinosad, and Fenitrothion (Sumithon) on the immature stages of the first and the second generations of the egg parasitoid T. evanescens They reported that no parasitoids emerged from parasitized eggs treated with CAPL-2. There was no emergence of the parasitoid treated with Lambdacyhalothrin, spinosad, and fenitrothion (Sumithon) one, two or four

42 71 days after parasitism. However, emergence was recorded with very low percentages when the eggs were treated 24 hrs before parasitoid emergence. Female percentage slightly decreased in the adults emerged from parasitized eggs treated with chemicals.

43 72 CHAPTER III MATERIALS AND METHODS The present investigation was conducted to study the performance of egg parasitoid Trichogramma chilonis (Ishii) under laboratory conditions at the Biological Control Laboratory, Department of Agricultural Entomology, Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth Dapoli, Dist. Ratnagiri during The details of materials used and methodologies adopted during the course of investigation are dealt under this chapter. 3.1 Mass production and maintenance of Corcyra cephalonica (Stainton) culture The nucleus culture of C. cephalonica in the form of eggs was procured from NBAII, Bangalore, and was mass produced and the culture was maintained in a separate room. Bold grains of sorghum were used as larval food. The bold grains of sorghum were milled in a domestic milling machine making two pieces of each grain and heat sterilized in hot air oven at C for 30 minutes to make free from any secondary infestation. Material was also treated with streptomycin 0.2 g per kg, to prevent the bacterial infection. Crushed, raw 250 g was added to one kilogram of sorghum and kept in pre-sterilized wooden trays (30 x 20 cm). Trays were arranged in a metal shelf. Ant wells were provided to avoid crawling of ants (Plate I). Each tray was inoculated with 1000 eggs of Corcyra (0-1 hr old) and thoroughly mixed to have uniform distribution of eggs in food material. These trays were secured with lid so as to avoid escape of either larvae or moths. The trays were kept undisturbed for 40 days.

44 73 After 40 days, the moths were collected by using mechanical moth collection device and transferred to the specially designed oviposition cages daily at 8.00 a.m. from each tray (Plate II). The collected moths were allowed to mate and lay eggs in separate egg laying cages prepared from plastic bucket (20 lit capacities) (Plate III). The eggs thus laid were collected in the next morning and cleaned to remove scales and other body parts of the moths, by rolling them on blotting paper. Cleaned eggs were further used to maintain the Corcyra culture and for undertaking various research aspects. 3.2 Mass production and maintenance of T. chilonis culture The nucleus culture of T. chilonis in the form of trichocard was procured from NBAII, Bangalore. Cards (15 x 8 cm) were prepared from white colour cardboard paper to maintain the Trichogramma culture in the laboratory throughout the experiment period (Plate IV). The eggs of Corcyra were used for the preparation of Trichocards. Cleaned eggs of C. cephalonica were exposed to Ultra Violet (U.V) radiation in a U.V chamber (Plate V) for 45 minutes at a height of 15 cm so as to kill the embryo. These eggs were used for preparing Trichocards. One cc eggs of Corcyra were sprinkled on each card smeared with a thin layer of pure gum arabic mixed with distilled water with 5 ml plastic vial having a perforated lid. A single layer of uniformly distributed Corcyra eggs was formed on the card. After proper drying, the individual egg card was introduced in glass bottle (15 25 cm) (Plate VI) having freshly emerged T. chilonis adults. Adults of T. chilonis were fed on 5 per cent honey solution in the form of a fine streak on the inner wall of the bottle. Adults were

45 74 allowed to parasitize Corcyra egg cards for 24 hours. Trichocards thus parasitized were shifted to separate plastic bottles. Each parasitized card was provided with the date of parasitization and proposed date of emergence of the parasitoid. The parasitized eggs turned black on the fourth day due to the deposition of black granules in the inner surface of the chorion. This was an indication of the development of parasitoid. The parasitoid emergence started after seventh day of parasitization. Hence, new cards were prepared after seven days for maintaining the pure culture, which further was utilized in various studies. 3.3 Mass production and maintenance of Spodoptera litura (Fab.) culture The initial culture of S. litura was obtained by collecting the larval stages from the infested crop like caster and mary gold grown in University Farm. The larvae then reared in cylindrical transparent glass jars (42 cm height; 30 cm diameter) (Plate VII) having piece of blotting paper at their bottom. Fresh tender leaves of caster were provided as food for larvae twice a day. The top of the glass jar was covered with muslin cloth secured firmly with rubber band. Fully developed last instar larvae were hand picked and transferred to glass jars having 7.5 cm thick layer of fine soil at the bottom. Moths emerged from pupae were used for building up of subsequent culture. Newly emerged male and female were confined into large glass jars for mating alongwith folded pieces of black blotting paper for egg laying. The freshly emerged male and female moths were confined to single glass jar in ratio of 2:3, respectively. Five such sets were maintained for egg laying. The inner side of each glass jar was lined with blotting paper to create favorable conditions for egg laying. A piece of circular blotting paper was placed at the bottom of each glass jar. Cotton swab soaked in 10 per cent sugar

46 75 solution was kept suspended in each glass jar as adult food. The cotton swab was changed periodically. The egg laid on blotting paper were collected by cutting away the piece of paper alongwith the egg mass and cleaned. The Cleaned eggs were further used to maintain the S. litura culture and for undertaking various research aspects. 3.4 Development of T.chilonis on C. cephalonica and S. litura Freshly laid, U.V irradiated 20 eggs of respective host s were pasted randomly on a small white paper card strip (4 x 3.5 cm) separately with the help of diluted gum which were then air dried and were served to 3 pre-mated freshly emerged females of T. chilonis from mass culture in a small glass vial (7.5 7 cm). After 24 h, the paper strip were removed and kept in a separate same size vial for further development of the parasitoid and following biological parameters of T. chilonis from respective host s trichocards were recorded separately Per cent parasitization Black coloured eggs were considered as the parasitized eggs, on the basis of which, per cent parasitization was determined Per cent adult emergence Out of total parasitized eggs, the number of parasitoids emerged were counted and per cent adult emergence was determined Sex ratio To determine sex ratio the emerged adults were first killed by keeping them under refrigerator at 0 0 C. From those killed adults, sexing was done by observing individual parasitoid under 10X hand lens based on the morphological characters, as given below. (Plate VIII).

47 76 Male: Male was darker and usually smaller than female, with black colour short, round abdomen. Antenna was distinct, long, more plumose, with black long bristles. Dorsum of thorax was brown with black tinge Female: Female was pale in colour and larger than male. Body was yellowish orange, with tapering abdomen. Antenna was short, not distinct, was less plumose with few short bristles Adult longevity Newly emerged five male and female adult parasitoids were kept individually in a small glass vial (7.5 x 7 cm) without food. The adult longevity was worked out from their emergence till death Total developmental period From first day of parasitization till death of adult was recorded to determine total developmental period. 3.5 Effect of host egg age on performance of T. chilonis Design : C.R.D Treatments : 13 Repetitions : 3 Table: 1 Different host egg age

48 77 Tr. No. Egg age (h) Tr. No. Egg age (h) T1 0-1 T T2 6-7 T T T T T T T T T T Approximately 1000 freshly emerged Corcyra moths were confined in oviposition cage and freshly laid eggs (0 to 1 h old) were immediately collected. They were aged further with an interval of 6 h from the time of egg laying as per the treatments mentioned earlier. Hundred eggs as per age were glued to white colour card paper strip (4 x 3.5 cm) which was then air dried and exposed to 5 freshly mated parasitoid females in a plastic vial (7.5 x 7 cm). They were exposed for 24 h and then the strip was removed and was kept in a separate same size vial for further development of the parasitoid. The per cent eggs parasitized and sex ratio was worked out as mentioned earlier. 3.6 Effect of different coloured trichocards on performance of T. chilonis

49 78 Design : C.R.D Treatments : 9 Repetitions : 3 Table: 2 Different Trichocard Colours Tr. No. Colour Tr. No. Colour T1 Red T6 Pink T2 Yellow T7 Brown T3 Green T8 Saffron T4 Black T9 White T5 Blue Different coloured strips of trichocard were offered to the parasitoid female in three different ways as given below No choice test Respective colour paper strip of size 4 x 3.5 cm with plane surface was prepared as colour trichocard strip. U.V sterilized hundred Corcyra eggs were glued on it with the help of gum and airdried. Individual colour strip was then exposed to 5 pre-mated females of T. chilonis in a separate glass vial (7.5 x 7 cm) (Plate IX) for 24 h. Next day, the cards were removed and kept under supervision till egg colour changed to black. Black colour eggs were considered as parasitized and per cent parasitization was worked out from number of eggs parasitized out of 100. Similarly, from

50 79 emerged adults, per cent female adults emerged and sex ratio was determined as mentioned earlier Free choice test (Open light) Respective colour paper strip of size 4 x 3.5 cm was glued with U.V sterilized hundred Corcyra eggs with the help of gum and airdried. The strips were glued in a line, vertically on the desiccator s inner wall (28 x 22 cm) with the help of cello tape (Plate X). Colours were placed randomly, taking care that two similar colours would not come nearby. All colour strips were then exposed to female of T. 100 pre-mated females for 24 h. Next day, the cards were removed and kept in individual plastic vial (7.5x 7 cm) under supervision till egg colour changed to black. Black colour eggs were considered as parasitized. Per cent parasitism was worked out from number of eggs parasitized out of 100. Similarly, from emerged adults, per cent female adults emerged and sex ratio was determined as mentioned earlier Free choice test (In dark condition) All the procedure was same except the way of exposure of the colour strips. Here the colour strips were exposed to Trichogramma females in a desiccator (28 x 22 cm). The strips were glued on the desiccator s inner wall with the help of cello tape. Colours were placed randomly, taking care that two similar colours would not come nearby. The whole assembly was then completely covered with a black cloth and was kept in total dark condition for 24 h. Remaining observations were recorded in a similar pattern as above. 3.7 Effect of low temperature storage on parasitized trichocard

51 80 Design : C.R.D Treatments : 9 Repetitions : 3 Table: 3 Storage period of parasitized trichocards at 15 0 C Tr. No Storage period (days) Temp ( 0 c) T T T T T T T T T Thirty trichocard strips of size 4 x 3.5 cm were marked on 15 x 8 cm cardboard paper accommodating 10-strips/paper. Hundred U.V. sterilized Corcyra eggs were glued on each strip and air-dried. They were then mass exposed to freshly emerged Trichogramma adults for 24 h. Individual strip was then carefully cut when eggs turned black (5 days after parasitization) and was kept in a separate plastic vial with secured cap (7.5 x 7 cm), which

52 81 further was marked for days to be kept in refrigerator with a marker pen. All such vials along with strips were kept in refrigerator at 5 th day of parasitization and were removed after specific storage period. Observations on per cent adult emerged was recorded to judge the effective storage period. 3.8 Relative toxicity of insecticides for T. chilonis Design : C.R.D Treatments : 10 Repetitions : 3 The insecticides were evaluated for their safety to the parasitoid in three different ways as follows. The treatment details are given in Table Contact toxicity to female T. chilonis Contact toxicity of insecticides (Table 4) was determined as per methodology suggested by Jalali and Singh (1993). A transparent plastic container (7.5 x 7 cm) with a tight lid was converted in to a pesticide-testing unit by cutting a small window on two sides and closing them with fine muslin cloth, glued with waterproof glue to avoid fumigant effect of insecticides and to provide aeration (Plate XI). Prepared solution of insecticide was sprayed on all inner sides of container and lid with an atomizer and both were then airdried thoroughly. In control treatment, only water spray was done. Twenty, newly emerged females were introduced inside this vial and exposed to treated surface freely (Plate XII). Five per cent honey was provided in the form of fine streak. The per cent adult mortality was recorded 24, 48 and 72 h after constant exposure. For the laboratory screening of different insecticides to test their safety to

53 82 adult female T. chilonis, the scores were assigned as follows based on per cent adult mortality. Sr.No. Particulars Per cent adult mortality (%) Score 1 Harmless Slightly harmful 50 to Moderately harmful 80 to Harmful >99 4 (Anon, 1994) Insecticide safety: Effect on egg parasitization White card paper strip (4 x 3.5 cm) with fresh, U.V sterilized 100 Corcyra eggs; glued with the help of diluted gum was prepared. It was air-dried thoroughly and dipped in respective insecticide solution for five seconds. In control the card paper strip was dipped in distilled water. Strips were again thoroughly air dried and then exposed to 5 freshly emerged, mated females for 24 h. After 24 h, stripes were removed and kept in another same size vials, individually. Number of eggs parasitized per treatment was judged as mentioned earlier. Based on reduction in parasitization compared to control, the insecticides were categorized into four categories as below

54 83 Sr.No. Particulars Reduction in parasitization over control (%) Score 1 Harmless < Slightly harmful 30 to Moderately harmful 80 to Harmful >99 4 (Baladandi et al., 2005) Effect on adult emergence Required number of white card paper strips (4 x 3.5 cm) with U.V sterilized 100 Corcyra eggs glued with the help of diluted gum were prepared and mass exposed to Trichogramma females for 24 h. After four days when the eggs changed their colour to black, the strips alongwith the parasitized eggs were dipped in respective insecticide solution for five seconds. Control was kept with water dipping of strips only. They were then air dried and individually kept in plastic vial (7.5 x 7 cm). Observations were recorded on per cent adult emerged from each treatment. Based on reduction in adult emergence as compared to control, the insecticides were categorized into four categories Sr.No. Particulars Reduction in adult emergence over control (%) Score 1 Harmless < Slightly harmful 30 to Moderately harmful 80 to Harmful >99 4 (Baladandi et al., 2005)

55 84 Table: 4 Details of insecticides treatments Sr. Treatments Conc. used Trade name source 1 Spinosad 45 SC Tracer 45 SC Dow Agro Science India Pvt. Ltd. 2 Triazophos 40 EC 0.05 Hostathion 40 EC Bayer Crop Science (India)Ltd. 3 Acephate 75 SP 0.1 Asataf 75 SP Bayer Crop Science (India)Ltd. 4 Lambda cyhalothrin 5 EC Warrior plus 5 EC Nagarjuna Agrichem Limited 5 Cypermethrin 25 EC Trofy 25 EC M/S United Phosphorus Ltd. 6 Azadirachtin 10,000 ppm Nemazol 1% EID Parry (India) Ltd. 7 Imidachloprid17.5 SL Confidor 17.8 SL Bayer Crop Science (India)Ltd. 8 Emamectin benzoate 5 SG Proclaim 5 SG Bayer Crop Science (India)Ltd. 9 Novaluron 10 EC Rimon 10 EC Indofil Chemical Company, Mumbai. 10 Water 43

56 85 CHAPTER IV RESULTS AND DISCUSSION The present investigations were undertaken to study the performance of egg parasitoid Trichogramma chilonis (Ishii) under laboratory conditions during the year in the Bio-control laboratory, Department of Agriculture Entomology, College of Agriculture, Dapoli (Maharashtra). The results are presented and discussed under the following headings. 4.1 Development of T. chilonis on Corcyra cephalonica (Stainton) and Spodoptera litura (Fab.) 4.2 Effect of host egg age on the performance of T. chilonis 4.3 Effect of different coloured trichocards on the performance of T. chilonis 4.4 Effect of low temperature storage on parasitized trichocards 4.5 Relative toxicity of insecticides for T. chilonis 4.1 Development of T. chilonis on C. cephalonica and S. litura The host influences the growth and survival of the developing parasitoids as well as the fecundity, longevity, developmental period and sex ratio etc., Thus one would expect improvement in their biological efficiency by rearing them on some suitable hosts. Hence to find out the egg parasitoid biological efficiency two hosts viz., C. cephalonica and S. litura were selected and following biological parameters were tested. The results are discussed under following sub headings Per cent parasitization

57 86 The data on per cent parasitization of Corcyra and Spodoptera eggs by T. chiolis females are presented in Table 5 and depicted graphically in Figure 1. The data revealed that among the host eggs offered, the parasitoid accepted eggs of both the hosts for parasitization. Among the hosts accepted, maximum and significantly superior per cent eggs parasitized was noticed on eggs of C. cephalonica (86.67 %) which was highly preferred for parasitization by the parasitoid and minimum per cent eggs parasitized was noticed on eggs of S. litura (76.67 %) which was least preferred by the parasitoid. Ram and Irulandi (1989) reported highest per cent egg parasitization in case of eggs of C. cephalonica (85.24 %) followed by C. partellus (77.47 %) and lowest in eggs of E. lunata (52.25 %). Selvaraj and Sundarababu (1995) also reported similar findings of highest per cent egg parasitization (78.40 %) by T. chilonis in eggs of P. operculella followed by per cent in C. cephalonica and lowest in eggs of S. litura (42.80 %) and H. armigera (33.40 %). Their findings also support the present results Per cent adult emergence The data on per cent adult emergence of T. chilonis on both the host eggs are presented in Table 5 and depicted graphically in Figure 2. The data revealed that the maximum per cent adult emergence was noticed on the eggs of S. litura (90.13 %) and followed by eggs of C. cephalonica (89.48 %) which was at par with S. litura.

58 87 Rathi and Ram (2000) recorded and per cent T. chilonis adult emergence when reared on C. cephalonica and H. armigera, respectively. Their findings also in line with the present results Per cent T. chilonis female adults emergence The data on per cent female adult emergence of T. chilonis on both the host eggs are presented in Table 5 and depicted in Figure 3. The data revealed that the maximum per cent female Trichogramma emergence was noticed on eggs of S. litura (67.35 %) and lowest on eggs of C. cephalonica (60.22 %). The per cent female Trichogramma emergence on S. litura was significantly superior over per cent female Trichogramma emergence on C. cephalonica. Rathi and Ram (2000) recovered higher female population from emerged adults of T. chilonis from the eggs of A. moorei (74.90 %) followed by E. vittella (73.30 %), H. armigera (71.50 %) and lowest (68.70 %) from eggs of C. cephalonica. Present findings also revealed the results as above Sex ratio (M : F) The data on sex ratio of T. chilonis reared on eggs of C. cephalonica and S. litura are presented in Table 5. The sex ratio was female biased on both host eggs. The maximum female parasitoid recovery was noticed from S. litura eggs (1:2.1) and less female parasitoid recovery was noticed from C. cephalonica eggs (1:1.53). Earlier work of Jalali et al. (1987) revealed that sex ratio in T. remus was 1:1.10 from S. litura, while 1:0.86 from eggs of C. cephalonica. Mehendale (2009) recorded that sex ratio in T. chilonis was 1:1.40 and 1:1.30 in S. litura and C. cephalonica eggs recpectively. Present findings also revealed higher females from eggs of

59 88 S. litura and lowest from C. cephalonica thus followed the results as above Adult longevity of male T. chilonis The data on adult longevity of male parasitoid emerged from both the host eggs are presented in Table 5 and depicted graphically in Figure 4. The results revealed significant difference in the longevity of emerged parasitoid males from eggs of both host. From eggs of S. litura the male parasitoid lived much longer (3.67 days) compared to shorter (2.83 days) from eggs of C. cephalonica. Rathi and Ram (2000) reported longer (5.6 days) male adult longevity on H. armigera and shortest (4.6 days) on C. cephalonica. Present findings also revealed longer male adult longivity from eggs of S. litura and shorter from C. cephalonica Adult longevity of female T. chilonis The data on adult longevity of femalr parasitoid emerged from both the host eggs are presented in Table 5 and depicted graphically in Figure 5. The results revealed significant difference in the longevity of emerged female parasitoid from eggs of both the hosts. Female parasitoids from eggs of S. litura lived much longer (5.33 days) compared to shorter (4.50 days) from eggs of C. cephalonica. Rathi and Ram (2000) reported longer (8.5 days) female adult longevity on H. armigera and shortest (7.8days) on C. cephalonica. Their findings also support the present results Total development period The data on total development period of parasitoid from both the host eggs are presented in Table 5 and depicted graphically in Figure 6. The results revealed significant difference in the total development

60 89 period of parasitoid on eggs of S. litura which was longest (11.17 days) and shortest (9.33 days) on the eggs of C. cephalonica. Rathi and Ram (2000) found that the development period was longer (11.2 days) on H. armigera eggs and shorter (9.3 days) on eggs of C. cephalonica. Present findings also revealed longer development period from eggs of S. litura and shorter from C. cephalonica. 4.2 Effect of host egg age on the performance of T. chilonis Among the different situations of the host egg in which the Trichogramma develops, host egg age play a vital role. To understand the role of host egg age in the performance of parasitoid, an experiment was conducted in the laboratory with different age Corcyra eggs (unsterilized). The results of the experiments are discussed under following heads Effect of host egg age on the parasitization potential T. chilonis The data on the per cent Corcyra eggs parasitized with varying ages by T. chilonis female are presented in Table 6 and depicted in Figure 7. Results revealed that freshly laid eggs of the age 0-1 h old were highly preferred by T. chilonis as evidenced by per cent parasitization, which was significantly superior over rest of the egg ages. This was followed by 6-7 h (88.67 %) and h (84.33 %) which were at par with h (83.00 %). Again this was followed by egg age h which recorded per cent parasitization. Up to h about 80 per cent parasitization was noticed. This means that egg age from 0-1 h to h were most suitable parasitization there after response of T. chilonis female decreased gradually as the

61 90 egg age advanced viz., h (65.33 %) and h (61.33 %). This was further followed by a decrease in per cent parasitization in egg age h (54.67 %), h (46.00 %) and h (41.33 %). Egg parasitization in further age eggs totally collapsed as evidenced by only per cent in h, 4.67 per cent in h and even no parasitization in h. From this experiment it was confirmed that freshly laid eggs were more vulnerable to the attack of egg parasitoid but as the host embryo advances in its development, such eggs were at all rejected by the parasitoid. This might be due to the behavioural response of the female parasitoid in selecting freshly laid eggs for oviposition. Earlier, Krishnamurti (1938) reported the host egg age effect on egg parasitoid in case of T. minutum on Corcyra eggs. Also Singh et al. (2001) in a study on effect of host egg age on T. exiguum and T. brasiliensis revealed that in case of fresh eggs the rate of parasitization was higher which declined as the eggs became older. Similar findings were also reported by Budhwanth et al. (2008). They reported that maximum parasitization (60.3%) was noticed in fresh eggs of 24 hrs old followed by 48 hrs old eggs (49.5%), while 72 hrs old eggs were less preferred (37.3%) by T. chilonis for parasitization. Mehendale (2009) reported egg age from 0-1 h to h old were highly preferred for parasitization. Thus, the present results are in accordance with all the above findings Effect of host egg age on female parasitoid emergence Development of the embryo of the egg parasitoid like Trichogramma mostly depends on nutrition they get in the host egg. Also the male embryo develops faster than female embryo and consumes more egg yolk in a short period. Thus host egg age is limiting factor in this sense. More yolk will be available in the fresh

62 91 eggs than in older eggs, which might decides the sex of the developing embryo. To confirm this, sexes were differentiated from the emerged individuals of the parasitoid from different age groups to determine per cent female emerged. The data are presented in Table 6 and depicted in Figure 8. The data revealed that egg age 0-1 h, 6-7 h and h produced 69.28, and per cent females, respectively which were at par with each other Further, egg age h, h and h produced 65.46, and per cent females respectively which were significantly superior over rest of the egg ages. Thereafter the per cent female emergence was declined drastically from h (59.24 %), h (56.10 %), h (50.75 %), h (43.53 %) and h (21.80 %). Further egg age h and h even could not produced significant adults and those emerged were all males. From the present findings it was revealed that lesser the egg age more was the female emergence. This might be due to the fact that in early stage of the host egg the host embryo remains in the embryonic development and once the egg is parasitized the parasitoid embryo starts fast development and utilizes the yolk very efficiently, which might provides greater chances of more females from such early age eggs. Another reason behind this may be as quoted by Wiackowski (1962). According to him the female selectivity for such fresh eggs was responsible where favorable conditions for development of parasitoid will be there and hence the female undergoes super parasitization, which leads to the selective elimination of male sex by the domination of females or due to the competition among the progenies. However, Singh et al. (2001) reported in case of T. exiguum that irrespective of host age, females were found to be predominant up to 60 to 61 h and was found to be above 75 per cent in all the age group from 0 to 1 to 48 to 49 h. However, in present study the female preponderance was

63 92 noticed only up to 24 to 25 h age and thereafter it declined drastically, which is at variance with Singh et al. (2001). Budhwanth et al. (2008) reported fresh eggs of 24 hrs could produce more female progeny of T. chilonis (52.9%) compared to old age of 72 hrs (48.7%). However, Mehendale (2009) reported maximum female recovery in egg age 0-1 h to h old. Thus, the above findings strongly support the present results Sex ratio (M : F) Based on the number of adults emerged, which were segregated into males and females and the sex ratio was worked out. The data on sex ratio are presented in Table 6. The data revealed that the sex ratio was female biased in egg age 0-1 h (1:2.25), 6-7 h (1:2.13) and h (1:2.04). This was followed by h (1:1.89), h (1:1.70) and h (1:1.54). Thereafter, the proportion of female progeny reduced drastically as revealed in egg age h (1:1.45), h (1:1.27) and h (1:1.03). As the egg age prolonged the sex ratio was male biased as revealed in egg age h (1:0.76) and h (1:0.27) and even no female adults noticed from egg age h and h. Wiackowski (1962) reported the female selectivity for fresh eggs was responsible where favourable conditions for development of parasitoid will be there and hence the female undergoes super parasitization, which leads to the selective elimination of male sex by the domination of females or due to the competition among the progenies. However, Singh et al. (2001) reported in case of T. exiguum that irrespective of host age, females were found to be predominant up to 60 to 61 h and was found to be above 75 per cent in all the age group from 0 to 1 to 48 to 49 h. However in present study the female preponderance was noticed only up to 42 to 43 h age and thereafter it declined drastically.

64 Effect of different coloured trichocard on the performance of T.chilonis Insects can detect different colour intensities to locate their host. To study the response of T. chilonis to different colours of trichocards, an experiment was conducted with nine different colours of trichocards in the laboratory. Three different conditions viz., no choice, free choice with open light and free choice with dark condition were given. The results are discussed under following heads Effect on parasitization potential No choice test Nine different colours of trichocards were offered to T. chilonis female individually with no other choice. The observations were recorded on per cent eggs parasitized on each colour background. The data on parasitization potential in no choice test are presented in Table 7 and depicted graphically in Figure 9. The results revealed that under no choice test most of the colours except saffron were preferred (Plate XIII). Among the various colours, egg parasitization recorded in white (88.33 %), yellow (87.33 %) and blue (86.67 %) were at par. Further, red (83.67 %), green (83.33 %) and brown (81.33 %) were also at par, followed by black (78.00 %) and pink (64.33 %). Saffron was the least preferred colour (51.00 %) Free choice open light To study the response of T. chilonis with all colours together, the colours were offered in an open light situation. The data on the per cent egg parasitized were recorded and are presented in Table 7 and illustrated in Figure 9. The data revealed that in free choice with open light condition, T. chilonis female perceived most of the colour wave lengths and hence found attempting egg parasitization. In almost

65 94 all colours, satisfactory and above per cent egg parasitization was noticed. This indicated that T. chilonis could perceive a broad range of colour wavelengths. Further it was revealed that green, yellow, blue and white colours with 88.67, 88.00, and per cent egg parasitization were superior and at par with each other followed by saffron with per cent parasitization. However brown (76.33 %) and red (75.33 %) were found at par with each other. Similarly, pink (68.67 %) and black (67.00 %) were also at par with each other. Thus, in free choice open light test, colours viz., green, blue, white and saffron supported the maximum egg parasitization (Plate XIII) Free choice dark condition To know the colour perception of T. chilonis under dark condition, an experiment with free colour choice but with dark condition was conducted. The data are presented in Table 7 and depicted in Figure 9. The data revealed that even though under total dark condition T. chilonis parasitized Corcyra eggs on all the colours of trichocards with almost same intensity except yellow and white (Plate XIII). Among the different colours T. chilonis preferred green (90.00 %), black (89.00 %), red (88.67 %) and brown (87.00 %) which were statistically at par and better than rest of all the colours. Further, brown was at par with saffron (84.67 %) while, saffron was at par with pink (82.33 %). Compared to above colours blue colour (77.00 %) supported lower per cent parasitization. Similarly, yellow (76.64 %) and white (74.00 %) were at par with each other. Thus, T. chilonis can search their host against a broad colour except blue and a faint background like yellow and white under dark condition Effect on female emergence No choice test

66 95 The data on the per cent female parasitoids emerged out of total adults emerged from respective colours of trichocard in no choice test are presented in Table 7 and depicted in Figure 10. The data revealed that maximum female parasitoid emergence was supported by green (68.21 %), white (66.87 %), brown (65.09 %) and yellow (64.75 %) which were at par. Similarly, white (66.87 %), brown (65.09 %) and yellow (64.75 %) were at par with red (63.63 %). Further, brown (65.09 %), yellow (64.75 %) and red (63.66 %) were at par with pink (62.02 %) and saffron (61.68 %). Similarly, pink (62.02 %) and saffron (61.68 %) were at par with blue (58.20 %). However, lowest per cent female emergence was noticed from blue (58.20 %) and black (55.74 %) which were at par with one another Free choice open light Regarding per cent female parasitoids emerged in free choice open light situation the data are presented in Table 7 and depicted in Figure 10. The data revealed that green (70.31 %), yellow (69.44 %), white (68.33 %) and saffron (67.10 %) were at par which supported highest per cent female emergence. Further, yellow (69.44 %), white (68.33 %), saffron (67.10 %) and brown (65.25 %) were at par. Again white (68.33 %), saffron (67.10 %) and brown (65.25 %) were at par and similarly saffron (67.10 %) was found at par with brown (65.25 %); blue (59.13 %) and red (57.91 %) were at par with each other. Black (51.67 %) and pink (45.01 %) colour supported the minimum per cent female emergence Free choice dark condition The data on the per cent female emerged in free choice with darkness are presented in Table 7 and Figure 10. The data revealed that under free choice dark condition green (68.25 %), red (65.55 %), white (65.42 %), saffron (65.33 %) and black (64.94 %) were at par

67 96 which supported higher per cent female emergence. Similarly red (65.55 %), white (65.42 %), saffron (65.33 %), and black (64.94 %) were at par and white (65.42 %), saffron (65.33 %), black (64.94 %) were with same line and saffron (65.33 %) at par with black (64.94 %) followed by brown (60.25 %) which was at par with blue (60.11 %). Pink (55.60 %) and yellow (50.20 %) supported the minimum per cent female emergence Sex ratio (M : F) No choice test The data on the sex ratio in no choice test are presented in Table 7. The data revealed that the sex ratio was with female preponderance in green colour (1:2.14) followed by white (1:2.02), brown (1:1.87), yellow (1:1.84), red (1:1.75), pink (1:1.66), saffron (1:1.64), blue(1:1.39) and black (1:1.28) Free choice open light The data on the sex ratio in free choice open light are presented in Table 7. The data revealed that the sex ratio was with female preponderance in yellow (1:2.27), white (1:2.19), saffron (1:1.93), brown (1:1.89), green (1:1.83), blue (1:1.45), red (1:1.41) and black (1:1.07), while male preponderance was in pink (1:0.89) Free choice dark condition The data on the sex ratio in free choice dark condition are presented in Table 7. The data revealed that the sex ratio was with female preponderance in green (1:2.16), saffron (1:1.99), red (1:1.90), white (1:1.90), black (1:1.85), pink (1:1.58), blue (1:1.54), brown (1:1.51) and yellow (1:1.02). From the overall results it was revealed that egg parasitoids like Trichogramma can actively search their host on varying background

68 97 colours as evidenced from all above three different situations in which nine different colours were displayed in front of parasitising female of T. chilonis. From the results it was revealed that green colour was preferred more as evidenced by higher rate of parasitization and female emergence at least in both the free choice tests. Earlier workers also reported this colour effect viz., Kadlubouski, (1970) who reported yellow colour to be most favourable for parasitization by T. embryophagum. Bhattacharya and Basit (1997) and Singh et al. (2001) found that colour of trichocard play an important role in parasitising efficacy of T. chilonis. Baitha and Sinha (2002) in a similar study with white, orange, yellow, red and green trichocard reported that significantly higher fecundity (number of eggs) was recorded on white (110) trichocard followed by green (86.80). Green trichocard supported maximum and significantly higher emergence of adult (76.29 %) and female (76.46 %) than on other colours. They realized that each colour influenced the adult emergence, which they attributed to hue of the colour. Comparing white and green colours, the per cent of oviposition was highest (88.64 %) on green trichocard in the first 3 days than on white trichocard (84.62 %). Further, they recommended white or green colour trichocard for mass multiplication of T. chilonis in laboratory. Bhattacharya et al. (2003) studied the effect of colour of cards used to fix C. cephalonica eggs on the behaviour and biological attributes of the parasitoid T. chilonis. Under multiple-choice conditions, the parasitization percentage varied significantly on differently colour egg cards. The maximum parasitization was observed on mint-green coloured egg cards (64.70 %), followed by jade green (56.75 %), golden yellow (54.25 %), deep orange (52.25 %), bus green (49.25 %), canary yellow (46.25 %) and white (40.50 %). The lowest parasitization was observed on chassis gray (11.50 %), followed

69 98 by dawn (14.50%) and wild lilac (15.75 %) coloured cards. The maximum emergence was on mint green cards (88.06 %), followed by golden yellow (88.04 %) and white (87.76 %). The lowest emergence was observed in wild purple egg cards (86.05 %). The highest female emergence was observed on white cards (72.63 %), while the lowest was on sky blue cards (71.09 %). In single choice conditions, the maximum fecundity was on mint green cards (87.50 %), followed by deep orange (87.33 %) and golden yellow (87.17 %). The maximum adult emergence was on white cards (87.00 %). Results of multiple and single choice experiments indicated that the insect preference was high for mint green egg cards, jade green, golden yellow, deep orange and white. However, Vishla et al. (2007) reported that T. chilonis showed maximum parasitism on yellow coloured cards (81.37 %), T. japonicum showed maximum parasitization on signle red (52.12 %) and T. brasiliensis on satin blue (82.00 %). Mehendale (2009) reported maximum per cent female parasitoid emergence in green colour. Thus, the above findings strongly support the present results. 4.4 Effect of low temperature storage on parasitized trichocards Trichocard prepared in the laboratory needs to be refrigerated when no demand or congenial conditions for release in the field are absent. Under such situation the question arises, up to how much period and temperature the trichocards could be effectively stored without hampering the adult emergence. To understand this, storage study in refrigerator for varying periods was undertaken. The data on per cent adult emerged at varying periods and 15 0 C are presented in Table 8 and depicted in Figure 11. The data revealed that maximum

70 99 per cent adult emerged were recorded at 5-day storage (88.67 %) followed by 10-day (85.33 %) which was at par with 5 days storage period. Further 15-day storage recorded per cent adult emergence, while 20 and 25 day storage recorded and per cent adult emergence, respectively remained at par, followed by 30-day storage with per cent adult emergence. Almost 50 per cent adult emergence was reduced at 35- day storage (50.67 %). Further reduction in adult emergence was noticed at 40 and 45- day storage with and 8.33 per cent, respectively. From the above results the impact of cold storage revealed that Trichocards could be stored effectively without much damage to the adult emergence upto 30 days at 15 0 C. Earlier work of Khosa and Brar (2000) on storage study with T. chilonis strain collected from different places revealed that at 8 to 10 0 C the mean per cent adult emergence was 96.2, 88.5, 85.1, 81.2, 73.2, 49.1, 33.6, 14.5 and 1.7 for a period of 1, 7, 15, 22, 29, 36, 43, 50 and 57 days, respectively. Similarly, Gupta and Bhardwaj (2002) also reported that storage of Trichogrammatoidea bactrae Nagaraja at 15 0 C for 10 days after 5 days of parasitization was appropriate.

71 100 However, Vishla et al. (2008) reported that the parasitized eggs of C. cephalonica stored at 10 C showed decreasing trend of parasitoid emergence with the increase in storage period. Mehendale (2009) reported that parasitized trichocards can be stored effectively up to 30 days at 15 0 C. Thus, the present findings are in line with that of above findings. 4.5 Relative toxicity of insecticides for T. chilonis Most of the time egg parasitoids like Trichogramma are used alongwith insecticides in the field. Trichogramma are highly sensitive to chemical pesticides. In view of this, a laboratory evaluation of some insecticides was undertaken to find out the safer insecticide through contact toxicity to female, insecticides safety to parasitization and adult emergence from already parasitized eggs of Corcyra. The results are discussed under following heads Contact toxicity of different insecticides to T. chilonis The per cent adult mortality at 24, 48 and 72 hours after treatment (HAT) was recorded. The results are discussed below Per cent adult mortality at 24 HAT The data on per cent adult mortality at HAT are presented in Table 9 and depicted graphically in Figure 12. The data revealed that no mortality was recorded in control (water spray). This was followed by azadirachtin ppm (0.004 %) and novaluron 10 EC ( %) with and per cent adult mortality, respectively. Further, emamectin benzoate 5 SG ( %) recorded per cent adult mortality. Remaining all the insecticides viz., spinosad 45 SC ( %), triazophos 40 EC (0.05 %), acephate 75 SP (0.1 %), lambda cyhalothrin 5 EC (0.003 %), cypemethrin 25 EC ( %) and imidachloprid 17.5 SL (0.005 %) recorded 100 per cent adult mortality within 24 HAT.

72 Per cent adult mortality at 48 HAT The data on per cent adult mortality at 48 HAT are presented in Table 9 and depicted graphically in Figure 12. The results revealed that as the exposure period increased, the per cent adult mortality aggravated. The data realized that the control recorded the lowest adult mortality of per cent. This was followed by azadirachtin (33.33 %), novaluron (48.33 %) and emamectin benzoate (77.00 %). All the remaining insecticides proved to be detrimental causing cent per cent adult mortality Per cent adult mortality at 72 HAT The data on per cent adult mortality at 72 HAT are presented in Table 9 and depicted graphically in Figure 12. The results revealed that control recorded per cent adult mortality while azadirachtin recorded % per cent adult mortality followed by novaluron which recorded % per cent adult mortality. All the remaining insecticides including emmamectin benzoate proved to be detrimental causing cent per cent mortality Per cent adult mortality (Pooled) The data on per cent adult mortality (Pooled) are presented in Table 9 and depicted graphically in Figure 12. Results of the pooled data of 24, 48 and 72 HAT revealed that control treatment recorded per cent adult mortality and remained significantly superior over rest of all the treatments. Among different insecticides, azadirachtin and novaluron remained at par with and per cent mortality, respectively. Further emamectin benzoate recorded per cent adult mortality. All the remaining insecticides were at par and produced 100 per cent mortality viz., spinosad, triazophos, acephate, lambda cyhalothrin, cypermethrin and imidachloprid.

73 102 Based on the observations from pooled data the ranking to the various insecticides given was as follows. 1 = Harmless (2% adult mortality) 2 = Slightly harmful (50 to 79% adult mortality) 3 = Moderately harmful (80 to 99% adult mortality) 4 = Harmful (>99% adult mortality) : No insecticide : azadirachtin and novaluron : emamectin benzoate : spinosad, acephate, lambda cyhalothrin, cypermethrin and imidachloprid From the above all results it was ascertained that there was no insecticide under testing totally safe to the parasitoid female. However, azadirachtin and novaluron were slightly harmful, while emamectin benzoate was moderately harmful and all the remaining insecticides were harmful. Different scientists have worked with different insecticides and with different species of Trichogramma and have different results and opinions about them. Among them Thakur and Pawar (2000) reported that neem-based pesticides and bio-pesticides were harmless, while endosulfan was slightly toxic. Charles et al. (2000) reported, based on LC50 values, spinosad and prophenophos were the most toxic compounds to T. exiguum adult females, followed by lambda cyhalothrin, cypermethrin, and thiodicarb. Prem et al. (2001) evaluated contact toxicity of different insecticides to adults of T. chilonis and revealed that deltamethrin ( %), malathion (0.05 %) and endosulfan (0.05 %) recorded cent

74 103 per cent mortality 24 h after exposure, while acephate (0.05 %) recorded per cent mortality. Control recorded per cent mortality. Boomathi et al. (2005) evaluated different botanicals and microbial pesticide mixtures against T. chilonis. The results revealed that Neem + Sweet flag and Neem + Sweet flag and 0.18 per cent, respectively registered up to per cent mortality 24 h after exposure. Among the microbial pesticides, spinosad (75 g a.i per ha), spinosad + HaNPV (75g a.i per ha + 1.5X10 12 POBs per ml, spinosad + spicturin + HaNPV (75g a.i per ha + 1.5X10 12 POBs per ml + 1lit per ha) caused cent per cent mortality 24 h after exposure as compared to per cent in endosulfan (0.07 %). Singh and Kaur (2005) reported that chlorpyriphos showed 100 per cent mortality within 15 minutes of exposure. Butter and Dhalival (2006) noticed that mortality of adults of T. chilonis was 1.33, 4.00 and per cent in , and ppm of Econeem, respectively. Thus, the present findings are supported by the above results Insecticides safety to egg parasitization Trichogrammatids in the field parasitize various lepidopteran eggs in different crops that might come in contact with the insecticides sprayed and may repel or kill the parasitising females reducing rate of egg parasitization. To ascertain this, an experiment with already treated Corcyra egg strips with respective insecticide was offered to T. chilonis female. The data on the per cent eggs parasitized are presented in Table 10 and depicted in Figure 13. The results revealed that significantly highest egg parasitization of per cent was recorded in control (water spray). Among the various

75 104 insecticides acephate and imidachloprid recorded and per cent parasitization, respectively and were at par. This was followed by emamectin benzoate and novaluron with and per cent eggs parasitized, respectively and were at par. Further, insecticides viz., azadirachtin, spinosad, triazophos and lambda cyhalothrin supported minimum parasitization of 38.33, 31.67, and per cent, respectively. Based on the data, ranking of the insecticides with respect to the parasitization of the pre-treated Corcyra eggs was allotted on the basis of per cent reduction in egg parasitization over control which was as below. 1 = Harmless (< 30 % reduction) 2 = Slightly harmful (30 to 79% reduction) 3 = Moderately harmful (80to99% reduction) 4 = Harmful (> 99% reduction) : acephate, imidacloprid, emamectin benzoate and novaluron : azadirachtin, spinosad and cypermethrin : triazophos and lambda cyhalothrin : No insecticide From these results it was clear that acephate and imidachloprid could equally favour greater per cent of egg parasitization followed by emamectin benzoate, novaluron. However, azadirachtin, spinosad and cypermethrin slightly reduced the paratization by T. chilonis. Earlier different workers conducted experiments with varying insecticides to see their effects on egg parasitization. Among them,

76 105 Kumar and Santharam (1999) revealed no significant adverse effect of imidachloprid on per cent parasitisation of T. chilonis. In another study, Prem et al. (2001) reported highest parasitization of per cent in acephate 0.05 per cent while lowest in endosulfan 0.05 per cent (2.90%), deltamethrin per cent (4.54%) and malathion 0.05 per cent (5.13%) as compared to per cent in control. Babasaheb et al. (2009) worked on effect of some newer insecticides alongwith microbials and a botanical on parasitization efficacy and they reported that neem oil 1.0% and lambda-cyhalothrin % reduced the parasitization of T. chilonis. Present results are strongly supported by all above findings Insecticides safety to adult emergence To understand the side effects of the insecticides on the adult emergence from already parasitized Corcyra eggs, an experiment was conducted by treating already parasitized eggs of Corcyra with the respective insecticide and the data on the per cent adults emerged were noted and are presented in Table 11 and depicted in Figure 14 The result of the experiment revealed that significant highest per cent adult emergence was recorded in control (94.00 %). Among the different insecticides tested azadirachtin recorded the maximum per cent adult emergence (83.67 %) followed by acephate (57.33 %). Insecticides viz., emamectin benzoate, novaluron, triazophos and cypermethrin recorded 38.33, 36.33, and per cent adult emergence, respectively. Moreover, imidachloprid (8.67 %) and lambda cyhalothrin (7.00 %) reported the minimum adult emergence and were at par. Spinosad was the most toxic insecticide

77 106 for adult emergence and caused cent per cent mortality of the developing Trichogramma adults within the eggs. From the data the categories allotted to different insecticides based on per cent reduction in adult emergence over control were as below. 1 = Harmless (< 30 % reduction) 2 = Slightly harmful (30 to 79% reduction) : azadirachtin : acephate, emamectin benzoate, novaluron, triazophos and cypermethrin 3 = Moderately harmful (80 to 99% reduction) 4 = Harmful (> 99% reduction) : imidachloprid and lambda cyhalothrin : spinosad Earlier Oznipar and Kornosor (1998) studied the effect of four different insecticides viz., monocrotophos SCW, cyfluthrin EC, thiodicarb DF and cypermethrin EC and they reported 58.71, 49.98, and per cent adult emergence from the parasitized host eggs, respectively. Similarly, Charles et al. (2000) reported that lambda cyhalothrin, cypermethrin and spinosad adversely affected emergence of Trichogramma from eggs of Helicoverpa zea (Boddie). Prem et al. (2001) reported mean per cent adult emergence in acephate 0.05 per cent (44.01 %), deltamethrin per cent (3.62 %), endosulfan 0.05 per cent (1.92 %), malathion 0.05 per cent (3.42 %) as compared to control (63.83 %). Babasaheb et al. (2009) observed that lambda-cyhalothrin % adversely affected the emergence of T. chilonis. The neem oil

78 107 did not adversely affect the development and emergence of T. chilonis. Mehendale (2009) studied the safer insecticides against T. Chilonis. He reported that no insecticide was found safe to developing adult Trichogramma inside the egg. Insecticides like triazophos 40 EC (0.05 %), acetamiprid 20 SP (0.004 %), acephate 75 SP (0.1 %), emamectin benzoate 5 SG (0.001 %), novaluron 10 EC ( %) and endosulfan 35 EC (0.075 %) were slightly harmful, imidacloprid 17.8 SL (0.005 %) as moderately toxic and fenobucarb 50 EC (0.1 %) as the most toxic one. Shoeb (2010) noticed that there was no emergence for the T. chilonis treated with lambda-cyhalothrin, spinosad, and fenitrothion one, two or four days after parasitism. However, emergence was recorded with very low percentages when the eggs were treated 24 hrs before parasitoid emergence. Present findings are in close agreement with all the above workers, stating that, all the insecticides except azdirachtin and acephate, reduced adult emergence drastically.

79 108 CHAPTER V SUMMARY AND CONCLUSION The experiment entitled Performance of egg parasitoid Trichogramma chilonis (Ishii) under laboratory conditions was undertaken in the Bio-control laboratory, Department of Agriculture Entomology, College of Agriculture, Dapoli (Maharashtra), during the year The results are summarized as below. 5.1 Development of Trichogramma chilonis (Ishii) on Corcyra cephalonica (Stainton) and Spodoptera litura (Fab.) Studies on the development of T. chilonis on C. cephalonica and S. litura revealed that maximum per cent parasitization was recorded on the eggs of C. cepholonica (86.67 %) and minimum per cent parasitization was recorded on the eggs of S. litura (76.67 %). The per cent adult emergence was found maximum on the eggs of S. litura (90.13 %) followed by C. cephalonica (89.48 %) which was at par with S. litura. Maximum per cent female parasitoid emergence was noticed from Spodoptera (67.35 %) eggs parasitized by T. chilonis and lowest per cent female emergence was noticed from Corcyra (60.22 %) eggs parasitized by T. chilonis. Sex ratio (M : F) of the emerged parasitoid progeny revealed that on Spodoptera eggs (1 : 2.10) maximum female recovery was noticed and less female parasitoid was noticed from C. cephalonica eggs (1 : 1.53). Male parasitoids longevity was longer (3.30 days) which were emerged from Spodoptera eggs and shorter (2.83 days) those emerged

80 109 from Corcyra eggs, while female parasitoid longevity was longer (5.33 days) which were emerged from Spodoptera eggs and found shorter (4.66 days) those emerged from Corcyra eggs. The total development period was also found longest (11.17 days) on eggs of Spodoptera and shortest (9.33 days) on eggs of C. cepholonica. From the above result it can be concluded that the development of T. chilonis was found superior over eggs of S. litura except in per cent parasitization on eggs of C. cepholonica. 5.2 Effect of host egg age on the performance of T. chilonis Effect of host egg age on the per cent parasitization by T. chilonis revealed that egg age from 0 to 1 h (93.33 %) to 24 to 25 h old (80.33 %) were highly preferred by the parasitoid female while late egg age from 24 to 25 h old onwards were less accepted and even no parasitization was seen in 72 to 73 h old eggs. Maximum per cent female parasitoids emerged from 0 to 1 h old eggs (69.28 %), followed by 6 to 7 h (68.62 %) and 12 to 13 h (67.19 %) while egg age 18 to 19 h (65.46 %), 24 to 25 h (63.07 %) and 30 to 31 h (61.26 %) as next effective egg ages. Further, the female emergence declined drastically as the egg age advanced. Sex ratio (M : F) was female biased in fresh eggs. Maximum females were emerged from egg age 0 to 1 h (1:2.25) followed by 6 to 7 h (1:2.13) and 12 to 13 h (1:2.04). As the egg age prolonged the sex ratio was male biased as revealed in egg age 54 to 55 h (1:0.76) and 60

81 110 to 61 h (1:0.27) and even no female adult noticed from egg age 66 to 67 h and 72 to 73 h. 5.3 Effect of different coloured trichocards on the performance of T. chilonis Effect of trichocard on the parasitization potential of T. chilonis evaluated in no choice, free choice open light and free choice dark, condition revealed that in no choice test highest per cent eggs parasitized were obtained in white (88.33 %), yellow (87.33 %) and blue (86.67 %). Saffron (51.00 %) supported the lowest per cent eggs parasitized. Under free choice open light condition, maximum per cent parasitization were obtained in green (88.67 %), yellow (88.00 %), blue (86.33%) and white (84.67 %), while saffron (82.33 %) as next effective colour. Free choice with dark condition recorded highest per cent parasitization in green (90.00 %) followed by black (89.00 %), red (88.67 %) and brown (87.00 %). Per cent female parasitoid emerged in different colour trichocards in above three conditions revealed that green colour supported the maximum females (68.21 %) followed by white (66.87 %) and yellow (64.75 %). However, lowest per cent female emergence was noticed from blue (58.20 %) and black (55.74 %) in no choice test. In free choice open light, again green colour supported maximum female parasitoid emergence (70.31 %) followed by yellow (69.44 %), white (68.33 %), saffron (67.10 %) and brown (65.25 %), while black

82 111 (51.67) and pink (45.01) colours supported the minimum per cent female emergence. Under Free choice dark condition maximum female emergence was noticed in green (68.25 %) followed by red (65.55 %), white (65.42 %), saffron (65.33 %) and black (64.94 %), while pink (55.60) and yellow (50.20) supported the minimum per cent female emergence. Sex ratio (M: F) in different colour trichocards under different situation revealed that under No choice test, green (1:2.14), white (1:2.02), brown (1:1.87) and yellow (1:1.84) supported more female preponderance, while blue(1:1.39) and black (1:1.28) supported less female preponderance. In free choice open light, yellow supported the highest females (1:2.27) followed by white (1:2.19), saffron (1:1.93), brown (1:1.89) and green (1:1.83), while 1:0.89 was the only male influenced sex ratio recorded in pink. Under free choice dark condition sex ratio was female influenced in green (1:2.16) followed by saffron (1:1.99), red (1:1.90), white (1:1.90) and black (1:1.85) while yellow (1:1.02) supported least female preponderance. Thus as far as mass production of Trichogramma under laboratory conditions is concerned the results of Free choice open light indicated that trichocard colours viz., green, yellow, blue followed by white supported higher parasitization. However, maximum per cent female parasitoid emergence was recorded only in green colour from all three situations and hence it can be concluded that paper of green colour would be most suitable for preparing trichocards.

83 Effect of low temperature storage on parasitized trichocards Effect of cold storage under refrigerated conditions at 15 0 C temperature for varying periods on 4 days after parasitization of trichocards revealed that per cent adult emergence was quite high at 5 days storage (88.67 %) followed by 10 days (85.33 %), 15 days (76.67 %), 20 days (65.33 %), 25 days (63.67 %) and 30 days (60.67 %). Thereafter, the adult emergence declined drastically. From the above results it was found that parasitized trichocards can be stored effectively up to 30 days at 15 0 C, four days after parasitization. 5.5 Relative toxicity of insecticides for T. chilonis Based on the results of the pooled data of the contact toxicity of different insecticides, it was observed that all the insecticides were unsafe to T. chilonis females. However, based on the ranking given, the rank 1 harmless (2% adult mortality) included no insecticides, rank 2 slightly harmful (50 to 79% adult mortality) included azadirachtin ppm (0.004 %) and novaluron 10 EC ( %), rank 3 moderately harmful (80 to 99% adult mortality) included only emamectin benzoate 5 SG ( %), while all remaining were given rank 4 harmful (>99% adult mortality). Thus from these results it was noticed that most of the insecticides were highly toxic and hence should be refrained from spraying, after T. chilonis has been released in the field, however, only azadirachtin ppm (0.004 %) and novaluron 10 EC ( %) being slightly harmful can be used with ease.

84 113 Insecticide safety to egg parasitization revealed that per cent reduction in egg parasitization over control indicated acephate 75 SP (0.1 %), imidachloprid 17.5 SL (0.005 %), emamectin benzoate 5 SG ( %) and novaluron 10 EC ( %) as harmless (Score 1 = < 30 % reduction in egg parasitization over control), azadirachtin ppm (0.004 %), spinosad 45 SC ( %) and cypermethrin 25 EC ( %) as slightly harmful (Score 2 = 30-79% reduction in egg parasitization over control) while, triazophos 40 EC (0.05 %) and lambda cyhalothrin 5 EC (0.003 %) as moderately harmful (Score 3 = 80-99% reduction in egg parasitization over control). No insecides recorded as harmful (Score 4 = > 99% reduction in egg parasitization over control). Based on the results of laboratory evaluation, further it could be suggested that insecticides like acephate, imidacloprid, emamectin benzoate and novaluron could be used safely before Trichogramma release in the field. Further, insecticedes like azadirachtin, spinosad and cypermethrin could be used before Trichogramma release in the field, while insecticides like triazophos and lambda cyhalothrin should not be used in the field, as they can disturb the parasitization. Effect of insecticides on adult emergence revealed based on the ranking given azadirachtin ppm (0.004 %) was found safe (Score 1 = < 30 % reduction in egg parasitization over control) to developing adults of Trichogramma inside the egg. Insecticides like acephate 75 SP (0.1 %), emamectin benzoate 5 SG ( %), novaluron 10 EC ( %), triazophos 40 EC (0.05 %) and cypermethrin 25 EC ( %) were slightly harmful (score 2 = 30-79% reduction in egg

85 114 parasitization over control), imidachloprid 17.5 SL (0.005 %) and lambda cyhalothrin 5 EC (0.003 %) as moderately harmful (Score 3 = 80-99% reduction in egg parasitization over control) and spinosad 45 SC ( %) recorded as harmful (Score 4 = > 99% reduction in egg parasitization over control). From the above laboratory results it could be understood that insecticide azadirachtin can be safely used in the field after release of Trichogramma. However insecticides like acephate, emamectin benzoate, novaluron, triazophos and cypermethrin can wisely used in the field 4-5 days after release of Trichogramma, while use of imidachloprid, lambda cyhalothrin and spinosad should strictly be avoided. The studies on parasitization potential and insecticides safety to T. chilonis were carried out under laboratory conditions in single trail. It is, therefore, felt necessary to conduct such trails for one more season under field conditions also.

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99 128 APPENDIX ABBREVIATIONS USED % : Per cent / : : At the rate < : Less than > : More than 0C : Degree celsius a.i. : active ingredient Anon. : Anonymous Bt : Bacillus thuringiensis CD : Critical difference C.R.D : Completely randomized design CASE : Custard apple seed extract cm : Centimeter DDT : Dichloro diphenyl trichloroethane Dist. : District EC : Emulsifiable concentrate et al. : and co-workers etc. : Etcetera g : Gram(s) Ha : Helicoverpa armigera (Hubner) ha : Hectare HAT : Hours after treatment h : Hours i.e. : That is IPM : Integrated pest management L : Litre LC50 : lethal concentration M.S. : Maharashtra state ml : Milliliter(s) mm 2 : Millimeter(s) square No. : Number NPV : Nuclear polyhedrosis virus NSKE : Neem seed kernel extract pp : Pages ppm : Parts per million PSE : Pongamia seed extract Pvt. Ltd. : Private limited S.Em ± : Standard error mean SG : Soluble granules

100 129 SL : Soluble liquid SP : Soluble powder Sr. : Serial Treat. US : : Treatments United states UV : Ultra violet Viz. : POBs : Namely Polyhedral occlusion bodies WSC : Water soluble concentrate

101 130

102 Per cent eggs parasitized No choice test Free choice with open light Free choice with dark condition Green Red Pink Brown Saffron White Yellow Blue Black Trichocard colours Figure : 9 Effect of Trichocard colour on egg parasitization

103 Per cent female parasitoid emerged No choice test Free choice with open light Free choice with dark condition 0.00 Green Red Pink Brown Saffron White Yellow Blue Black Trichocard colours Figure : 10 Effect of Trichocard colour on female parasitoid emergence

104 Per cent adult mortality Mortality at 24 HAT Mortality at 48 HAT Mortality at 72 HAT pooled 0.00 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 Treatments Figure : 12 Contact toxicity of different insecticides to Trichogramma chilonis (Ishii) female adults

105 Per cent adult emerged Storage period (Days) 45 Figure : 11 Effect of cold storage on adult emergence from parasitized trichocards

106 Per cent eggs parasitized T1 T2 T3 T4 T5 T6 T7 T8 T9 Treatments T10 Figure : 13 Insecticides safety Effect on egg parasitization

107 Per cent adult emerged T1 T2 T3 T4 T5 T6 T7 T8 T9 Treatments T10 Figure : 14 Insecticides safety Effect on adult emergence

108 137 Per cent parasitized C. cephalonicaa S. litura Host insects Figure : 1 Effect on parasitization potential Per cent adult emerged C. cephalonica S. litura Host insects Figure : 2 Effect on adult parasitoid emergence per cent female parasitoid emerged C. cephalonica S. litura Host insects Figure : 3 Effect on female parasitoid emergence

109 138 Male adult parasitoid longevity (Days) C. cephalonica S. litura Host insects Figure : 4 Effect on male parasitoid longevity (Days) Female adult parasitoid longevity (Days) C. cephalonica S. litura Host insects Figure : 5 Effect on female parasitoid longevity (Days) Total developmental period (Days) C. cephalonica S. litura Host insects Figure : 6 Effect on total development period of parasitoid (Days)

110 Per cent eggs parasitized Egg age (h) Figure : 7 Effect of host egg age on egg parasitization by Trichogramma cholonis (Ishii) Per cent female parasitoid emerged Egg age (h) Figure : 8 Effect of host egg age on female parasitoid emergence

111 Plate VIII : Male and Female adult of Trichogramma chilonis (Ishii) 140

112 141 Plate I : Mass production and maintenance of Corcyra culture Plate II : Corcyra moth collection using mechanical moth collection device

113 142 Plate III : Oviposition cages for Corcyra female Plate IV : Trichocards produced from nucleus culture

114 143 Plate V : UV chamber for egg irradiation Plate VI : Mass culture of Trichogramma chilonis (Ishii) in glass jars with irradiated eggs

115 Plate VII : Mass production of Spodoptera culture 144

116 145 No choice test Free choice open light test Free choice dark test Plate XIII : Effect of Trichocard colour

117 146 Plate XI : Plastic container modified for testing contact toxicity to different insecticides to Trichogramma adult Plate XII : Insecticide testing unit Contact toxicity

118 147 Plate IX : Effect of trichocard colour on parasitization No choice test Plate X : Effect of trichocard colour on parasitization Free choice open light test