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(Orthoptera: Gryllotalpidae) with the Entomopathogenic Fungus, Beauveria Bassiana (Balsamo) Vuillemin

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(Orthoptera: Gryllotalpidae) with the Entomopathogenic Fungus, Beauveria Bassiana (Balsamo) Vuillemin ABSTRACT THOMPSON, SARAH RACHEL. Biological control and behavioral studies of mole crickets (Orthoptera: Gryllotalpidae) with the entomopathogenic fungus, Beauveria bassiana (Balsamo) Vuillemin. (Under the direction of Rick L. Brandenburg.) The objectives of this research conducted with the entomogenous fungus, Beauveria bassiana, include: a) to determine if a behavioral response occurs when mole crickets are exposed to conidia b) to measure the viability of conidia when applied to turfgrass and c) to evaluate the efficacy of various strains and rates of the fungus against mole crickets in a laboratory topical bioassay. All three studies focused on two strains of B. bassiana propagated and formulated by JABB of the Carolinas, Inc. (Pine Level, NC), DB-2 and 10- 22. A commercially available strain, GHA formulated as BotaniGard ES, from Emerald Bioagriculture Corporation (Butte, MT) was also evaluated. Greenhouse studies at North Carolina State University (Raleigh, NC) to determine behavioral changes in response to the fungus were conducted during the fall and winter of 2001-2002 and the spring and summer of 2003 using tawny and southern adult mole crickets. Significant variations in behavior were observed in containers treated with strain DB-2 and bifenthrin, indicating avoidance responses associated with these control agents. Field research to evaluate conidial viability was conducted on Bermudagrass Cynodon dactylon ( L.) plots at Sea Trails Golf Course (Sunset Beach, NC) in August 2002 and Tri- State Sod Farm (Newton Grove, NC) in June 2003. Strains DB-2 and 10-22 were significantly more viable than BotaniGard in the 2003 study but all three strains were able to persist in the environment up to 28 days after application in both field studies. 10-22 showed the best performance in both field tests as well as after day 7 in an accompanying greenhouse trial. The laboratory topical bioassay was conducted in August 2003 on tawny and southern mole cricket nymphs to determine the most efficacious strain and rate of B. bassiana. There were significant differences in survival functions for all fourteen treatments (four rates of each of the three B. bassiana strains as well as two control treatments). BotaniGard caused the greatest mortality in the shortest amount of time after exposure. The highest rate tested, 108 conidia per cricket, appeared to have the most considerable effect for all three strains as measured by total percent mortality and LT25. Results from these studies emphasize the importance of strain selection for the use of B. bassiana as a biological control agent for mole crickets. An appropriate isolate that maintains viability, effectively targets the host, and dose not elicit behavioral responses from the crickets will have the most potential as an efficacious control agent for these serious turfgrass pests. BIOLOGICAL CONTROL AND BEHAVIORAL STUDIES OF MOLE CRICKETS (ORTHOPTERA: GRYLLOTALPIDAE) WITH THE ENTOMOPATHOGENIC FUNGUS, BEAUVERIA BASSIANA (BALSAMO) VUILLEMIN by SARAH RACHEL THOMPSON A thesis submitted to the Graduate Faculty of North Carolina State University in partial fulfillment of the requirements for the Degree of Master of Science ENTOMOLOGY Raleigh 2003 APPROVED BY: ___________________________________ ______________________________ _______________________________________ Chairman of Advisory Committee BIOGRAPHY Sarah Rachel (Devereux) Thompson was born the fourth of six children on 18 August 1977 in Baltimore, Maryland. She received her elementary and secondary education in Parkton, Maryland, graduating from Hereford High School in 1995. Sarah attended University of Maryland in College Park, graduating in 1999 with a Bachelor of Science degree in Secondary Science Education with an emphasis in Biology. After completing this degree, she taught sixth grade general science at Eagles Landing Middle School in Boca Raton, Florida for two years. The author began the pursuit of her Master of Science degree in July 2001 under the supervision of Dr. Rick L. Brandenburg. On 31 August 2002, she married Bryan Keith Thompson in Baltimore. ii ACKNOWLEDGMENTS The North Carolina Turfgrass Foundation and the Center for Turfgrass Environmental Research and Education provided generous financial support for this research. I am indebted to these two organizations and the Department of Entomology at North Carolina State University for giving me the opportunity to complete this degree. I thank Dr. Rick Brandenburg for taking a chance on me. He is a truly great mentor who lives his life the way I hope I someday can live mine, committed equally to his professional and personal lives. He has always made himself available to me when I need advice or a friend to talk to and for that I thank him. Drs. Jim Arends and Wayne Brooks provided guidance, time, and humor and I am grateful for having them as members of my committee. Their critique of this thesis was invaluable. The support of all three of these great entomologists is greatly appreciated. In Dr. Brandenburg’s lab, I would like to acknowledge Brian Royals, Brenda Watson, Christie Hurt, and Dr. Peter Hertl for their assistance and support over the past two and a half years. I thank them in advance for the next two. From the Department of Statistics, I thank Drs. Jason Osborne and Cavell Brownie for their help with the statistical analysis of my data. Without the cooperation and assistance of Steve White at The Lakes Country Club (Boiling Springs Lakes, NC), Ronny Wilson at Oyster Bay Golf Links (Sunset Beach, NC), Terry Vassey (formerly) at Sea Trails Plantation (Sunset Beach, NC), and Clark Wooten at Tri- iii State Sod Farm (Newton Grove, NC), my research would have not been possible. A special thanks to Bill Black at JABB of the Carolinas for his advice, time, and friendship. And, most importantly, to my husband, Bryan- words cannot express my gratitude for your support, love, and friendship always. Thank you and I love you. iv TABLE OF CONTENTS Page LIST OF TABLES………………………………………………………………………....viii LIST OF FIGURES………………………………………………………………………….ix INTRODUCTION…………………………………………………………………………....1 LITERATURE REVIEW………………………………………………………………….....3 Description and Distribution of Mole Crickets…………………………………..…..3 Biology and Ecology of Scapteriscus vicinus and S. borellii……………………..….5 Mole Cricket Damage………………………………………………………..……….7 Conventional Mole Cricket Control……………………………………………….....9 Biological Control of Mole Crickets…………………………………………..……12 Larra spp. (Hymenoptera: Sphecidae: Larrinae)………………………......13 Ormia depleta Wiedemann (Diptera: Tachinidae)………………………....15 Steinernema and Heterorhabditis spp. (Nematoda: Rhabditida)………..…16 Beauveria bassiana (Balsamo) Vuillemin……………………………….....21 References Cited………………………………………………………………...….28 TUNNELING RESPONSES OF MOLE CRICKETS (ORTHOPTERA: GRYLLOTALPIDAE) TO THE ENTOMOPATHOGENIC FUNGUS, BEAUVERIA BASSIANA (BALSAMO) VUILLEMIN…………………………………….…..….40 Abstract………………………………………………………………………….…..41 Introduction……………………………………………………………………..…...42 Materials and Methods…………………………………………………………..….44 v Propogation of Fungal Isolates……………………………………………...44 Collection of Mole Crickets……………………………………………...….44 Behavioral Studies…………………………………………………...……...45 2001-2002 Tests………………………………………………...…...46 2003 Tests……………………………………………………...……47 Statistical Analysis……………………………………………………..…....48 Results ……………….………………………………………………………....…...48 Mortality………………………………………………………………….....49 New Surface Tunneling…………………………………………………......49 Vertical and Horizontal Tunnels………………………………………..…...50 Cricket in Top Layer………………………………………………………...50 “Edging”…………………………………………………………..………...50 Discussion…………………………………………………………………………...51 References Cited……………………………………………………………….…....54 CONIDIAL VIABILITY AND EFFICACY OF BEAUVERIA BASSIANA (BALSAMO) VUILLEMIN FOR MOLE CRICKET (ORTHOPTERA: GRYLLOTALPIDAE) CONTROL IN TURFGRASS……………………………………………….……...64 Abstract……………………………………………………………………………...65 Introduction………………………………………………………………………….66 Materials and Methods……………………………………………………………...68 Propagation of Fungal Isolates…………………………………………..….68 Application of Fungal Conidia to Turfgrass……..…………………..……...69 2002 Tests………………………………………………………...…69 vi 2003 Tests……………………………………………………….......70 Conidial Suspension Preparation…………………………………………....71 Viability Assessment using Fluorescence Microscopy……………………..71 Collection of Mole Crickets……………………………………………..….72 Treatment of Mole Crickets with Fungus………………………………..…73 Incubation of Cadavers……………………………………………………..74 Statistical Analysis……………………………………………………….....75 Results and Discussion………………………………………………………...……75 Conidial Viability……...…………………………………………………....75 Topical Bioassay………………………………………………………..…..80 References Cited………………………………………………………………...…..84 CONCLUSIONS..………………………………………………………………………......99 vii LIST OF TABLES TUNNELING RESPONSES OF MOLE CRICKETS (ORTHOPTERA: GRYLLOTALPIDAE) TO THE ENTOMOPATHOGENIC FUNGUS, BEAUVERIA BASSIANA (BALSAMO) VUILLEMIN: 1. F values, degrees of freedom, and P values for control treatments in greenhouse behavioral studies conducted in 2001-2002 and 2003 (N=90)……………………...57 2. Mean area of surface tunnels produced after 24 hours by adult mole crickets in response to different treatments in greenhouse behavioral studies conducted in 2001- 2002 and 2003 (N=10)………………………………………………………………58 CONIDIAL VIABILITY AND EFFICACY OF BEAUVERIA BASSIANA (BALSAMO) VUILLEMIN FOR MOLE CRICKET (ORTHOPTERA: GRYLLOTALPIDAE) CONTROL IN TURFGRASS: 1. Mean percentage conidial viability at location 2 (Sea Trails Plantation, Sunset Beach, NC) 1, 2, 3,
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