Differential nymphal development of Blattella asahinai and B. germanica and their hybridization potential (Blattodea: Ectobiidae) by Madison Knox Peterson A thesis submitted to the Graduate Faculty of Auburn University in partial fulfillment of the requirements for the Degree of Master of Science Auburn, Alabama August 8, 2020 Key words: peridomestic pest, life cycle, reproduction, instar, clustering, development Copyright 2020 by Madison Knox Peterson Approved by Xing Ping Hu, Chair, Extension Specialist Professor Arthur G. Appel, Professor Katelyn A. Kesheimer, Assistant Professor & Extension Specialist Abstract The Asian cockroach, Blattella asahinai (Blattodea: Ectobiidae), is the closest relative of the German cockroach, B. germanica (L.), and has become a peridomestic nuisance pest in the southeastern United States. However, many basics aspects of the biology of B. asahinai, such as nymphal development and temperature thresholds have not yet been studied. The objectives of this project were to detail the nymphal development and developmental responses to temperature of B. asahinai as compared to B. germanica and examine the hybridization potential of these two species, as well as behavioral and morphological characteristics of the hybrids. In the following experiments, through measurements of nymphs, B. asahinai was estimated to have six or seven instars during nymphal development, and these results were subsequently confirmed through direct observation. Across six constant temperatures ranging from 10°C to 35°C, B. germanica had shorter nymphal development periods, fewer instars, and higher survivorship than B. asahinai. These developmental differences illuminated possible advantages that B. germanica and B. asahinai have in their respective environments. Both hybrid crosses were successfully performed, and the resulting hybrids appeared to have inherited intermediate wing morphology, allowing for flight in some hybrids. The results from these experiments will be useful in creating targeted management methods for B. asahinai based on unique biological traits that allow it to thrive in its outdoor habitat. 2 Acknowledgments I am grateful to my advisers Dr. Hu and Dr. Appel for their support during my time at Auburn. Their advice and encouragement have allowed me to persevere in the midst of difficulties. I am grateful to my committee member Dr. Kesheimer not only for her help in the preparation of this thesis, but also for the valuable extension opportunities that she has coordinated that I have been able to partake in. I also thank Mrs. Marla Eva for her invaluable assistance with supplies and colony maintenance. I am grateful to Dr. Snoddy and Dr. Wu for their generous provision of materials that contributed to this research. I thank my lab mates Gökhan Benk, Seun Oladipupo, Patrick Thompson, and Richard Murphy for their support, encouragement, and advice. I also thank the Department of Entomology and Plant Pathology faculty and staff for their support. I thank my parents, Nancy Dunlap and Michael Dunlap, for their dedication to my academics and career goals, as well as their sound guidance. Most importantly, I thank my Creator, who is always good, and who has always directed my paths. 3 Table of Contents Abstract ......................................................................................................................................... 2 Acknowledgments......................................................................................................................... 3 List of Tables ................................................................................................................................ 6 List of Figures ............................................................................................................................... 8 Chapter 1: Introduction and Literature Review .......................................................................... 10 Chapter 2: Instar determination of Blattella asahinai with digital measurements using Gaussian mixture models and number of cercus annuli ............................................................................. 36 Introduction ..................................................................................................................... 37 Materials and Methods .................................................................................................... 40 Results ............................................................................................................................. 46 Discussion ....................................................................................................................... 50 References ....................................................................................................................... 61 Chapter 3: Differential survival and development of Blattella asahinai and B. germanica (Blattodea: Ectobiidae) at six constant temperatures .................................................................. 80 Introduction ..................................................................................................................... 80 Materials and Methods .................................................................................................... 84 Results ............................................................................................................................. 88 Discussion ....................................................................................................................... 93 References ..................................................................................................................... 101 Chapter 4: When German cockroaches fly?: investigating the interspecific hybrids of Blattella asahinai and B. germanica (Blattodea: Ectobiidae) ................................................................ 114 Introduction ................................................................................................................... 115 4 Materials and Methods .................................................................................................. 117 Results ........................................................................................................................... 119 Discussion ..................................................................................................................... 121 References ..................................................................................................................... 124 5 List of Tables Chapter 2: Table 1. Mean digital and caliper pronotal length and width for each of six B. asahinai mixed sex instars determined using cercal annuli with results from paired t-tests ...................................... 67 Table 2. Mean pronotal length and width for six instar clusters identified by “mclust” analysis of 1,422 mixed sex B. asahinai nymphs ......................................................................................... 68 Table 3. Mean pronotal length and width by sex for the six instar clusters of 734 male and 688 female B. asahinai nymphs from “mclust,” using natural log-transformed pronotal data ......... 69 Table 4. Pronotal length and width means for six instars of 1,282 mixed sex B. asahinai nymphs based on dorsal cercal annuli ...................................................................................................... 70 Table 5. Pronotal length and width means of instars by sex from 650 male and 632 female B. asahinai nymphs determined from counting dorsal cercal annuli .............................................. 71 Table 6. Brooks-Dyar growth ratios for the six instars by sex calculated by dividing the mean length or width of each instar by the preceding instar ................................................................ 72 Chapter 3: Table 1. Cumulative mean time (d) and standard deviation to complete each instar of B. germanica and B. asahinai, as well as total nymphal development, at 20°C, with n in parentheses. ............................................................................................................................... 106 Table 2. Cumulative mean time (d) and standard deviation to complete each instar for six replicates of B. germanica and B. asahinai, as well as total nymphal development, at 25°C, with n in parentheses. ........................................................................................................................ 107 Table 3. Cumulative mean time (d) and standard deviation to complete each instar for six replicates of B. germanica and B. asahinai, as well as total nymphal development, at 30°C, with n in parentheses. ........................................................................................................................ 108 Table 4. Cumulative mean time (d) and standard deviation to complete each instar and total nymphal development for six replicates of B. germanica at 35°C, with n in parentheses ....... 109 Table 5. Linear regression models, results of ANOVA, lower development thresholds (°C), and degree days required for completion of each instar 1-7, as well as total nymphal development of B. asahinai, based on development at 20°C, 25°C, and 30°C .................................................. 110 6 Table 6. Linear regression models, results of ANOVA, lower developmental thresholds (°C), and degree days required for completion of each instar 1-6, as well as total nymphal development of B. germanica,