|UFDC Home||myUFDC Home | Help|
This item has the following downloads:
1 UNDERSTANDING AND IMPROVING THE RISK ANALYSIS PROCESS FOR APPROVING THE IMPORTATION AND RELEASE OF ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS INTO THE UNITED STATES EVALUATION OF CURRENT METHODOLOGIES AND LESSONS LEARNED FROM BIOLOGICAL CONTROL RESEARCH By OULIMATHE PARAISO A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2011
2 2011 Oulimathe Paraiso
3 To my family, especially my mother Alhaja Mulikat Paraiso
4 ACKNOWLEDGMENTS My sincere thanks to my supervisors: Drs. Moses Kairo and Stephanie Bloem for their academic and professional guidance. I would also like to thank members of my graduate committee (Drs. James Cuda, Stephen Hight, Norman Leppla, Michael Olexa, and Marcia Owens) for their assistance. I would like to thank the College of Engineering Sciences, Technology and Agriculture, Florida Agricultural and Mechani cal University (FAMU), the Department of Entomology and Nematology at University of Florida, and the United State Department of Agriculture Animal and Plant Health Inspection ServicePlant Protection and Quarantine for providing me with financial assistanc e and the opportunity to pursue my doctoral degree. This research project would not have been possible without the assistance of several scientists and support staff from the U.S. Department of AgricultureAgricultural Research ServiceCenter for Medical, Agricultural, and Veterinary Entomology in Tallahassee, FL. In particular, I would like to thank Dr. Stephen Hight for making field and laboratory experiments possible and also Christopher Albanese, John Mass, Michael Getman, and Shalom Benton (FAMU) for field and laboratory assistance. I would also like to thank Dr. Stuart Reitz for his statistical assistance and help in data analysis. In addition, I would like to thank Dr. James Carpenter, Susan Drawdy, and Robert Caldwell from the USDA ARS Crop Protection and Management Research Unit Tifton, Georgia for their statistical and technical assistance. I would also like to show my appreciation to my colleagues at the Center for Biological Control (FAMU). Finally, I would like to thank my mother, sister, and brother for their endless moral support.
5 TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES .......................................................................................................... 11 LIST OF FIGURES ........................................................................................................ 1 3 LIST OF ABBREVIA TIONS ........................................................................................... 15 ABSTRACT ................................................................................................................... 17 CHAPTER 1 INTRODUCTION .................................................................................................... 19 2 LITERATURE REVIEW .......................................................................................... 22 Negative Impacts of Invasive Species .................................................................... 22 The Importance of Biological C ontrol in Invasive Species Management ................. 23 Examples of Successful Biological Control Programs ............................................ 25 The Risk Analysis Process for Entomophagous Biological Control Agents ............ 26 Risks Associated with the Importation and Release of Entomophagous Biological Control Agents .............................................................................. 26 Pest Risk Analysis ............................................................................................ 27 Permitting Process ........................................................................................... 32 Risk Communication Framework during the Importation of Entomophagous Biological Control Agents .............................................................................. 33 Risk Communication during Pest Risk Analysis ............................................... 33 Improving Risk Communi cation in Governmental Agencies ............................. 34 Approaches to Communicating Risk ................................................................ 35 Mental Models Approach within Organizations ................................................. 36 Conditions for Using the Mental Models Approach ........................................... 37 Practical Application of Pest Risk Analysis into Biological Control Research ......... 37 Selecting a Successful Entomophagous Biological Control Agent ................... 37 Biological characteristics ............................................................................ 38 Functional response and numerical response ............................................ 38 Host suitability and preferences ................................................................. 39 Example of NonTarget Impacts: Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), a Spreading Pest in the U.S. ................................. 41 Inundative Biological Control as a Pest Management Strategy for Cactoblastis cactorum ................................................................................... 43 Assessing the Risk of Releasing a NonSpecific Egg Parasitoid to Control Cactoblastis cactorum in North America ....................................................... 44 Trichogramma (Hymenoptera: Chalcidoidea: Trichogrammatidae) .................. 45
6 Trichogramma pretiosum (Riley) ................................................................ 45 Trichogramma fuentesi Torre ..................................................................... 46 3 COMPARISON OF REGULATORY PROCEDURES FOR THE IMPORTATION AND RELEASE OF ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS IN EIGHT COUNTRIES HOW IS IT DONE? ........................................................ 49 Methodology ........................................................................................................... 51 Selection of Countries ...................................................................................... 51 Collation of Information .................................................................................... 52 Comparative Process ....................................................................................... 52 Results .................................................................................................................... 53 Australia (AU) ................................................................................................... 53 Designation of authority, laws and regulatory requirements ...................... 53 General acceptance of precaution ............................................................. 53 Documentary responsibility of importer ...................................................... 54 Australian regulatory process for importation and release of entomophagous BCAs (Figure 31) ........................................................ 54 Reviewing process/consultation ................................................................. 55 Responsibility of NPPO during and following release of BCA .................... 55 Canada (CAN) .................................................................................................. 55 Designation of authority, laws and regulatory requirements ...................... 55 General acceptance of precaution ............................................................. 56 Documentary responsibility of importer ...................................................... 56 Canadian regulatory process for importation and release of entomophagous BCAs (Figure 32) ........................................................ 56 Reviewing process/consultation ................................................................. 57 Responsibility of NPPO during and following release of BCA .................... 57 European and Mediterranean Region .............................................................. 57 Switzerland (SW) ....................................................................................... 59 Designation of authority, laws and regulatory requirements ...................... 59 General acceptance of precaution ............................................................. 60 Documentary responsibility of importer ...................................................... 60 Swiss regulatory process for importation and release of entomophagous BCAs ............................................................................ 60 Reviewing process/consultation ................................................................. 61 Responsibility of NPPO during and following release of BCA .................... 61 United Kingdom (UK) ................................................................................. 61 Designation of authority, laws and regulatory requirements ...................... 61 General acceptance of precaution ............................................................. 61 Documentary responsibility of importer ...................................................... 62 United Kingdom regulatory process for importation and releas e of entomophagous BCAs ............................................................................ 62 Reviewing process/consultation ................................................................. 62 Responsibility of NPPO during and following release of BCA .................... 63 India (IN) ........................................................................................................... 63
7 Designation of authority, laws and regulatory requirements ...................... 63 General acceptance of precaution ............................................................. 63 Documentary responsibility of import er ...................................................... 63 Indian regulatory process for importation and release of entomophagous BCAs ............................................................................ 64 Reviewing process/consultation ................................................................. 64 Re sponsibility of NPPO during and following release of BCA .................... 64 Mexico (MX) ..................................................................................................... 65 Designation of authority, laws and regulatory requirements ...................... 65 General acceptance of precaution ............................................................. 65 Documentary responsibility of importer ...................................................... 65 Mexican regulatory process for importation and release for entomophagous BCAs (Figure 33) ........................................................ 65 Reviewing process/consultation ................................................................. 66 Responsibility of NPPO during and following release of BCA .................... 66 New Zealand (NZ) ............................................................................................ 66 Designation of authority, laws and regulatory requirements ...................... 66 General acceptance of precaution ............................................................. 67 Documentary responsibility of importer ...................................................... 67 New Zealand regulatory process for importation and release of entomophagous BCAs (Figure 34) ........................................................ 67 Reviewing process/consultation ................................................................. 68 Re sponsibility of NPPO during and following release of BCA .................... 68 United States (U.S.) ......................................................................................... 68 Designation of authority and law/regulatory requirements ......................... 68 General acceptance of precaution ............................................................. 69 Documentary responsibility of importer ...................................................... 69 U.S. regulatory process for importation and release of entomophagous BCAs (Figure 3 5) ................................................................................... 69 Reviewing process/consultation ................................................................. 70 Responsibility of NPPO during and follow ing release of BCA .................... 71 Discussion .............................................................................................................. 71 4 RISK COMMUNICATION DURING THE IMPORTATION AND RELEASE OF ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS IN THE U.S.IS THERE ROOM FOR IMPROVEMENT? ................................................................. 84 Materials and Methods ............................................................................................ 88 Results .................................................................................................................... 91 Response Rate and Respondent Characterization .......................................... 91 Importance of Risk Communication .................................................................. 91 Risk Communication Framework ...................................................................... 91 Frequency and Sources of Risk Communication .............................................. 92 Goals of Risk Communication .......................................................................... 93 Respondent Satisfaction with Risk Information and Interactions ...................... 93
8 Need for more Guidance Documents ............................................................... 94 Public Involvement ........................................................................................... 94 Discussion .............................................................................................................. 94 5 COLLABORATIVE RISK ASSESSMENT DURING THE PERMITTING PROCESS OF ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS A BETTER PROCESS? ........................................................................................... 107 Materials and Methods .......................................................................................... 111 Committee of Experts ..................................................................................... 111 Development of an Expert Conceptual Model and Interview Protocol ............ 111 Comparison of Mental Models for DecisionMakers and Stakeholders .......... 112 Results and Discussion ......................................................................................... 112 Conceptual Expert Model ............................................................................... 112 Comparison of Mental Models for DecisionMakers and Stakeholders by Categories ................................................................................................... 113 Risk analysis process .............................................................................. 113 Stakeholder participation ......................................................................... 114 Risk c ommunication ................................................................................. 115 External review process and selection of expert group members ............ 116 Decision Making Process ............................................................................... 117 Approval for import to federal quarantine facility ...................................... 117 Approval for environmental release ......................................................... 117 Final decision ........................................................................................... 118 Summary .............................................................................................................. 118 6 EGG PARASITOIDS ATTACKING CACTOBLASTIS CACTORUM (LEPIDOPTERA: PYRALIDAE) IN NORTH FLORIDA .......................................... 127 Materials and Methods .......................................................................................... 130 Data Analy sis ........................................................................................................ 132 Results .................................................................................................................. 133 Discussion ............................................................................................................ 137 7 TRICHOGRAMMA FUENTESI A NEWLY DISCOVERED POTENTIAL BIOLOGICAL CONTROL AGENT OF CACTOBLASTIS CACTORUM EVALUATION OF BIOLOGICAL PARAMETERS ................................................. 146 Materials and Methods .......................................................................................... 147 Rearing Procedures and General Methods .................................................... 147 Effect of Presence and Type of Diet on Female Parasitoid Longevity ............ 148 Influence of T. fuentesi Female Age on Percent Parasitism ........................... 148 Effect of Female Mating Status on Percent Parasitism .................................. 149 Influence of Host Age on Percent Parasitism ................................................. 149 Statistical Analysis .......................................................................................... 150 Results .................................................................................................................. 150
9 Effect of Presence and Type of Diet on Female Longevity ............................. 150 Influence of Female Parasitoid Age and Mating Status on Percent Parasitism ................................................................................................... 151 Influence of Host Age on Percent Parasitism ................................................. 151 Discussion ............................................................................................................ 152 8 TRICHOGRAMMA FUENTESI EVALUATION OF FUNCTIONAL AND NUMERICAL RESPONSE .................................................................................... 162 Materials and Methods .......................................................................................... 163 Rearing Procedures ....................................................................................... 163 Functional and Numerical Response Experiments ......................................... 164 Statistical Analysis .......................................................................................... 165 Results .................................................................................................................. 166 Discussion ............................................................................................................ 167 9 TRICHOGRAMMA FUENTESI BEHAVIORIAL NOTES AND HOST SUITABILITY ........................................................................................................ 174 Materials and Methods .......................................................................................... 177 Trichogramma Rearing Procedures ............................................................... 177 Developmental and Reproductive Biology ...................................................... 177 Host Finding Behavior Study .......................................................................... 178 Host Specificity Tests ..................................................................................... 179 Non targ et host species selection ............................................................ 179 No choice tests ........................................................................................ 180 Statistical Analysis .......................................................................................... 180 Results .................................................................................................................. 180 Developmental and Reproductive Biology ...................................................... 180 Host Finding Behavior .................................................................................... 181 Host Suita bility Experiment ............................................................................. 181 Discussion ............................................................................................................ 182 10 CONCLUSIONS ................................................................................................... 191 APPENDIX A INTERVIEW INITIAL LETTER .............................................................................. 197 B INTERVIEW PRE NOTICE LETTER .................................................................... 198 C INTERVIEW BOOKLET ........................................................................................ 200 D EXAMPLES OF MENTAL MODELS FOR THE PERMITTING PROCESS FOR ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS IN THE U.S. ............... 209 E REMINDER NOTICE ............................................................................................ 210
10 F QUESTIONNAIRE ................................................................................................ 211 G INSTITUTIONAL REVIEW BOARD APPROVAL LETTER ................................... 218 LIST OF REFERENCES ............................................................................................. 219 BIOGRAPHICAL SKETCH .......................................................................................... 248
11 LIST OF TABLES Table page 2 1 Selection criteria of the centrifugal phylogenetic method for choosing test plants to determine host range of weed biological control agents ...................... 47 3 1 Criteria used in count ries examined in the pest risk analysis comparative study ................................................................................................................... 73 3 2 Comparison of general acceptance of precaution during pest risk an alysis and decisionmaking during permitting for entomophagous biological control agents in 8 countries .......................................................................................... 74 3 3 Comparison of documentary responsibilities of importer, prior to import, for pest risk analysis and decisionmaking during permitting for entomophagous biological control agents in 8 countries ............................................................... 75 3 4 Comparison of communication and reporting processes during pest risk analysis and decisionmaking during importation and release of entomophagous biological control agents in 8 countries .................................... 76 3 5 Comparison of reviewing and consultation processes for pest risk analysis and decisionmaking during permitting process for entomophagous biological control agents in 8 countries ............................................................................... 77 3 6 Comparison of responsibilities of the National Plant Protection Organi zation before, during and following release of biological control agent in 8 countries ... 78 4 1 Questionnaire ..................................................................................................... 99 4 2 Summary of questions and responses obtained from 5 categories of biological control stakeholders. ......................................................................... 101 5 2 Characterizing the mental models of decision makers and stakeholders during the permitting process of entomophagous biological control agents in the U.S. ............................................................................................................ 121 6 1 Sites surveyed in North Florida for egg parasitoids of Cactoblastis cactorum eggsticks on Opuntia spp. and additional informat ion on moth oviposition preference. ....................................................................................................... 140 6 2 Number of Cactoblastis cactorum eggsticks collected, lost in the field, examined in the laboratory, and number of eggs per eggstick SE at different sites in North Florida for different oviposition periods. ........................ 142
12 6 3 Location and date parasitized Cactoblastis cactorum eggstick was collected, identity of parasitoid species, number of eggs per eggstick, number of parasitized eggs, number of parasitoids emerged, female ratio, and parasitism level of egg parasitoids attacking C. cactorum in North Florida. ...... 143 7 1 The influence of age and mating st atus on number of Cactoblastis cactorum eggs parasitized by Trichogramma fuentesi ..................................................... 157 7 2 Influence of Cactoblastis cactorum egg age on parasitization by Trichogramma fuentesi ..................................................................................... 158 8 1 Parasitism of C. cactorum eggs at different densities by T. fuentesi ................ 171 8 2 Linear regressions of the proportion of C. cactorum eggs parasitized by T. fuentesi with increasing densities of eggs ........................................................ 171 9 1 Time (seconds) and rate per minute allocated by female Trichogramma fuentesi, over a 60 min observation period, for each ovipositional related behavior when associated with Cactoblastis cactorum eggs. ........................... 185 9 2 List of Lepidoptera species developed for complete host specificity testing of Trichogramma fuentesi ..................................................................................... 187 9 3 Parasitism (Mean S.E.) of potential host species by Trichogramma fuentesi 190
13 LIST OF FIGURES Figure page 2 1 Life stages of Cactoblastis cactorum .................................................................. 48 3 2 Canadian permitting process for entomophagous biological control agents ....... 80 3 3 Mexican permitting process for entomophagous biological control agents ......... 81 3 4 New Zealand permitting process for entomophagous biological control agents ................................................................................................................. 82 3 5 U.S. permitting process for entomophagous biological control agents as implemented in 2007 .......................................................................................... 83 4 1 Expert model of the permitting process used by USDA APHISPPQ in 2007 .. 102 4 2 Distribution of respondents to Question 1: In which group will you categorize yourself? .......................................................................................................... 103 4 3 Different model choices of pest risk analysis structure presented in questionnaire .................................................................................................... 103 4 4 Distribution of respondents to Question 4: How often do you communicate risk in the context of your profession? ............................................................. 104 4 5 Distribution of respondents to Question 6: From which entity(ies) do you receive information pertaining to risks associated with importation of BCAs and what is the rel ative importance of each source? ....................................... 104 4 6 Distribution of respondents to Question 11: Rank the following key goals of the risk communication process during the importation of entomophagous BCAs in order of importance ........................................................................... 105 4 7 Distribution of respondents to Question 8: How would you rate your level of satisfaction with the ris k communication message/information that you receive from USDA APHIS PPQ pertaining to the importation of entomophagous BCAs? .................................................................................. 105 4 8 Distribution of respondents to Question 9: How would you rate your level of satisfaction with the risk communication exchanges/interactions that you receive from USDA APHIS PPQ pertaining to the importation of entomophagous BCAs? .................................................................................. 106 4 9 Distribution of respondents to Question 12: How effective is USDA APHISPPQ in fulfilling each risk communication goal during the importation o f entomophagous BCAs? .................................................................................. 106
14 5 1 Novel risk assessment process for the permitting process for importation and release of entomophagous BCAs ..................................................................... 125 6 1 Locations surveyed for egg parasitoids of Cactoblastis cactorum in North Florida .............................................................................................................. 144 6 2 Ms. Paraiso surveying cactus plant for Cactoblastis cactorum eggsticks ......... 145 6 3 Parasitized Cactoblastis cactorum egg in an eggstick on Opuntia cactus pad 145 7 1 Non parasitized (left) Cactoblastis cactorum eggsticks glued on paper strip next to glued parasitized eggs (right) in eggsticks by Trichogramma fuentes i. 159 7 2 Rearing settings for Trichogramma fuentesi using Cactoblastis cactorum as host eggs .......................................................................................................... 1 59 7 3 Arrangement used to increase the relative humidity in rearing cultures of Trichogramma fuentesi ..................................................................................... 160 7 4 A regression showing the influence of Cactoblastis cactorum host egg age (1 20 days) on level of parasitism by Trichogramma fuentesi ......................... 161 8 1 Petri dishes set up to test functional response of individual Trichogramma fuentesi female parasitoids to various densities of Cactoblastis cactor um ....... 172 8 2 Functional response of Trichogramma fuentesi to different numbers of Cactoblastis cactorum egg densities ............................................................... 172 8 3 The number of Trichogramma fuentesi emerged per parasitized eggs (mean S.E.) with different Cactoblastis cactorum egg dens ities. .............................. 173 9 1 Methodology for selection of nontarget species based on Kuhlmann et al. 2006 ................................................................................................................. 186
15 LIST OF ABBREVIATION S APHIS Animal and Plant Health Inspection Service ARS Agricultural Research Service AQIS Australian Quarantine and Inspection Service BA Biosecurity Australia BCA(s) Biological Control Agent(s) BCRC Biological Control Review Committee CFIA Canadian Food and Inspection Agency CSIRO Commonwealth Scientific and Industrial Research Organization DAFF Department of Agriculture, Fisheries and Forestry DEH Department of Environmental and Heritage EA Environmental Assessment EBCA(s) Entomophagous Biological Control Agents EIS Environmental Impact Statement EPBC act Environment Protection and Biodiversity Conservation act FAO (United Nations) Food and Agriculture Organization F ONSI Finding of No Significant Impact IOBC International Organization for Biological Control IPPC International Plant Protection Convention ISPM International Standard for Phytosanitary Measures NBCI National Biological Control Institute NAPPO North Americ an Plant Protection Organization NEPA National Environmental Policy Act NPPO National Plant Protection Organization OPL Ontario Plant Laboratory
16 PHD Plant Health Division PPA Plant Protection Act PPQ Plant Protection and Quarantine PPQS Directorate of Pla nt Protection, Quarantine and Storage PRA Pest Risk Analysis QEL Quarantine Entomology Laboratory RA Risk Assessment RC Risk Communication RM Risk Management RSPM Regional Standard for Phytosanitary Measures SPS Sanitary and Phytosanitary Agreement TAG Technical Advisory Group USDA United States Department of Agriculture WTO World Trade Organization
17 Abstract of Dissertation Presented to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the D egree of Doctor of Philosophy UNDERSTANDING AND IMPROVING THE RISK ANALYSIS PROCESS FOR APPROVING THE IMPORTATION AND RELEASE OF ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS INTO THE UNITED STATES EVALUATION OF CURRENT METHODOLOGIES AND LESSONS LEARNED FROM BI OLOGICAL CONTROL RESEARCH By Oulimathe Paraiso May 2011 Chair: Moses T.K. Kairo Co Chair: Stephanie Bloem Major: Entomology and Nematology Classical biological control is a strategy to manage invasive pests. Despite many success stories involving the use of entomophagous biological control agents (BCAs), concerns have been raised about potential negative environmental impacts. T he current risk analysis process for entomophagous BCAs in the U.S. is considered by some to be subjective and often arbitrar y. Th e research undertaken in this dissertation developed a modified risk analysis framework to improve the importation and release of entomophagous BCAs. The aim of this revised procedure wa s to increase involvement and trust by stakeholders. In an effort to apply risk analysis concepts into research, a model example of a potential entomophagous BCA was examined. A case study was explored using the naturally occurring egg parasitoid, Trichogramma fuentesi Torre (Hymenoptera: Trichogrammatidae) att acking Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), a serious non native pest of Opuntia spp. in North Florida. In order to implement a safe and effective biological control program, biological and ecological characteristics of T. fuentesi were assessed. O ur research showed that one
18 to two day old mated T. fuentesi females should be used to increase percent parasitism in the field However, the correlation between the number of egg parasitized and host egg densities was weak suggesting that T. fuentesi would not provide a significant level of control of this pest in the field. Comparison of percent parasitism of C. cactorum eggs with preferred hosts showed a relatively low level of parasitism when the wasp attacked the invasive moth (11%) than when native Lepidoptera were attacked (26 75%) In addition, results suggested that inu n dative releases of T. fuentesi could potentially impact native cactus moth and butterfly eggs The findings from this study demonstrated that T. fuentesi is not a good candidate for augmentative biological control against C. cactorum Results from our research study provided a better understanding of the information necessary to improve the pest risk analysis for the importation and release of entomophagous BCAs in the U.S Inclusion of recommendations developed from the conceptual analysis and experimental studies into decisionmaking processes will improve implementation and safety of biological control programs in the U.S.
19 CHAPTER 1 INTRODUCTION Classical biological control is an important component of an integrated pest management strategy against invasive species. Classical biological control refers to the use of introduced natural enemies against nonnative pests (Eilenberg et al. 2001) Desp ite several examples of successful pest control using entomophagous biological control agents (BCAs) (van Lenteren et al. 2006) there are increasing concerns from the scientific community on the possible negative environmental and economic impacts associa ted with their importation and release (Howarth 1983; Simberloff & Stiling 1996; Louda et al. 2003) Many consider the current risk assessment process for entomophagous BCAs in the U.S. to be subjective and often arbitrary. In 2007, Interviews were condu cted to characterize how phytosanitary decisions wer e made during the permitting process for entomophagous BCAs by the U S Department of Agriculture (USDA), Animal and Plant Health Inspection Service (APHIS), Plant Protection and Quarantine ( PPQ ) the Nati onal Plant Protection Organization (NPPO) for the U.S., and to identify issues related to the risk assessment process that could be improved. Concerns about the potentially irreversible nontarget impacts that might result from the importation and release of entomophagous BCAs have resulted in increasingly stringent regulatory requirements by the U.S. NPPO. Despite numerous scientific publications on the potential risks associated with importation and release of BCAs there still are divergent opinions among regulators, researchers, environmentalists, and the general public on way s to appropriately manage these potential risk s. One approach that could be used to reduce this divergence might be through the implementation of a comprehensive and effective risk
20 communication process T his study examined communication habits of stakeholders involved in biological control and characterized how phytosanitary decisions we re communicated to them by decisionmakers working for the U.S. NPPO. We used the mental models approach to examin e how stakeholders perceived risks and underst oo d the associated scientific literature and we explored possible ways of providing a better understanding of factors that affect ed the permitting process for entomophagous BCAs. The second part of the dissertation focused on the evaluation of potential risks associated with entomophagous BCAs under laboratory conditions. The assessment of potential impacts is a crucial step during the risk assessment process. Cactoblasti s cactorum (Berg) (Lepidoptera: Pyralidae) is a serious pest of Opuntia spp. (prickly pear cactus) in North America. Presence of C. cactorum in the U.S. is of great concern because of its potential adverse impacts to ecological systems and to native and e ndangered Opuntia spp. particularly in the southwestern U.S. Geographical expansion of C. cactorum is also a threat to Opuntia spp. biodiversity and agricultural production in Mexico. Interest in the natural enemies of C cactorum has increased since the moth was found in Florida in 1989. Previous surveys of natural enemies in Argentina identified egg parasitoids in the family Trichogrammatidae as potential BCAs for C. cactorum Trichogramma wasps are widely distributed egg parasitoids used against majo r lepidopteran pests in greenhouse and field settings. Consequently, a study was conducted in North Florida to identify and to assess occurrence of egg parasitoids attacking this invasive moth in its new homeland. In addition, to further understand the i nteractions of the parasitoids and the pest, this research examined the following
21 biological parameters in the laboratory: parasitoid functional and numerical response, behavioral characteristics, and host specificity of Trichogramma fuentesi Torre (Hymen optera: Trichogrammatidae). The last part of the dissertation uses the findings from the conceptual analysis and the laboratory experiments to discuss the potential for inundative releases of T. fuentesi against C. cactorum in the U.S.
22 CHAPTER 2 LITERATU RE REVIEW Negative Impacts of Invasive Species The intentional introduction of several nonnative species of plants and animals to the U.S. such as corn, wheat soybeans and cattle has greatly benefited American agriculture, wh ereas unintentional introductions of many species have been the source of significant economic and environmental damage [e.g. Brazilian pepper tree ( Schinus terebinthifolius Raddi, Anacardiaceae) and Zebra mussels ( Dreissena polymorpha Pallas, Dreissenidae)] (A ndersen et al. 2004). One of the main concerns of the U.S. National Plant Protection Organization (NPPO) is the increasing rate of unintentional introductions of non native species that in many instances become invasive (Waage 2001). The introduction of these nonnative species without their natural enemies indirectly contributes to their invasiveness (Waage 2001). Concerns over the impact of i nvasive species on the U.S. economy have resulted in the development of new policies and action groups. For i nstance, in 1999 President Bill Clinton issued an E xecutive O rder for the allocation of US $28 million for creat ion of an Interagency Invasive Species Council to address the threat of invasive species (Pimentel et al. 2005). The Councils principal mission was to develop a strategy to fight the spread of nonnative species (Pimentel et al. 2005). In 2005, it was estimated that the annual economic cost of nonnative invasive species in the U.S. was almost US $120 billion per year (Pimentel et al. 2005). Arthropod pests alone cause a 13% overall reduction in crop yields. In economic terms, this reduction represents about US $33 billion in lost crop production annually (USBC 2001). Just 40% of these introduced pests account ed for a n annual total economic loss of US $13 billion (Pimentel et al.
23 1999). About US $500 million are spent each year applying pesticides to control crop pests (Pimentel et al. 1999) and another US $1 5 b illion are spent each year to manage entomophagous pests of lawns, gardens, and golf courses in the U.S. (Pimentel et al. 2005). In addition to their negative economic effects, invasive species have been recognized as a threat to the conservation of biodiversity (Convention on Biological Diversity, article 8h). Invasive species may cause permanent changes to ecological communities by shifting the arrangement and abundance of native species (Andersen et al. 2004). Almost half of the current species listed as threatened or endangered in the U.S. are considered to be at risk because of the potential predatory or competiti ve behaviors of nonnative species (Wilcove et al. 1998). Moreover, the presence of invasive species has been documented as a major constraint to reforestation, water management, and recovery of degraded lands in devel oping countries (Waage 2001). Invasive pest species are a threat to sustainable development ( Abate et al. 2000 ; Waage 2001; K airo 2005) A s ustainable approach is based on the management of natural resources to meet current human needs while maintaining t he earths capacity to meet the needs of future generations. At the social level, t here has been an increasing demand for green alternatives to chemical control of invasive species ( Charudattan 2001; Pim entel et al. 2005). The Importance of Biological C ontrol in Invasive Species Management Biological control can be a key component of an integrated pest management strategy against invasive species. Classical biological control is defined as the intentional introduction of an exotic natural enem y (e.g. insects, mites, nematodes, pathogens) that results in establishment of the enemy and, as consequence, effects
24 permanent control of the target pest (Eilenberg et al. 2001). There is an increas ed interest in the use of biological control by the U S D epartment of A griculture (USDA) because of the imminent withdrawal of several major pesticides and insecticides from the market (Charudattan 2001). Furthermore, development and registration of new pesticides involves high costs. Meanwhile, government al institutions have been mandat ing a reduction of chemical pesticide usage in food production (Charudattan 2001). For instance, the extensive use of herbicides for weed management resulted in the emergence of herbicideresistant weed biotypes. The economic and environmental impact s of herbicideresistant crops and naturally resistant weeds generated a need for pest management alternatives to unilateral chemical controls (Charudattan 2001). In addition, t here has been opposition from the general public toward excessive use of insecticides or development of genetically altered food crops (e.g. herbicidetolerant transgenic crops) to control invasive species ( Charudattan 2001 ; Pim entel et al. 2005). Successful biological control programs directly benefit cons umers because they usually have minimal impacts on the environment Pest control using biological control agents ( BCAs ) presents a safer alternative to pesticide use in food production systems (Charudattan 2001). The use of biological control minimizes pesticide impacts on people and the environment (Hoddle 2004). At the social level, biological control programs are a sustainable solution for developing countries and resourcepoor farmers because of biological control s intrinsic characteristics ( e.g., safe to humans, envi ronmentally friendly, and self sustaining) ( Abate et al. 2000 ; Waage 2001; K airo 2005)
25 Examples of Successful Biological Control Programs There are many examples of s uccessful biological control programs against weed and arthropod pests (Julien 1992; Julien & Griffiths1998) Infestations of purple loosestrife ( Lythrum salicaria L. Lythraceae), a rhizomatous perennial introduced to North America from Eurasia and Africa, was managed by the release of Galerucella pusilla Duftschmidt G calmariensis Duftschmidt (Coleoptera: Chrysomelidae) and Hylobius transversovittatus Goeze (Coleoptera: Curculionidae) ( Hight et al. 1995; Blossey et al. 1994; Landis et al. 2003). M anagement of the expansion of the Australian tree Melaleuca quinquenerv ia (Cav.) S.T. Blake (Myrtaceae) in Florida by th re e BCAs was another successful example of weed biological control (Serbesoff King 2003) Melaleuca had become one of Floridas most invasive weeds since its introduction in the 1880s (Fairchild 1947; Morton 1966; Balciunas 1990; Laroche & Ferriter 1993). Various scientific publications report the success of biological control programs against arthropod pests (van Lenteren et al. 2006). For instance, in Africa, the cassava mealybug [ Phenacoccus manihoti Matile Ferrero ( Hemiptera: Pseudococcidae) ] was effectively controlled by a parasitic wasp, Apoanagyrus lopezi (DeSantis) (Hymenoptera: Encyrtidae) imported from South America ( Herren & Neuenschwander 1991; Hammond et al. 1992). Cassava ( Manihot esculent a Crantz Euphorbiaceae) native to South America, and the preferred host of the cassava mealybug, is a staple food for millions of Africans (CGIAR 2008). An economic study showed that this biological control program generated a benefit of US $149.00 for every dollar invested (Norgaard 2006).
26 In North America, the positive impacts of entomophagous BCAs have been thoroughly documented. In 1888, the ladybird beetle, Rodolia cardinalis (Mulsant) (Coleoptera: Coccinellidae) was introduced into California t o control the c ottony cushion scale Icerya purchasi Maskell (Hemiptera: Margarodidae) (DeBach 1973). The cottony cushion scale was severely infest i ng California citrus groves causing orchard yield to decrease dramatically. The biological control project which cost only about US $1500, saved the California citrus industry millions of dollars (Caltagirone & Doutt 1989). C ontrol of the alfalfa weevil Hypera postica ( Gyllenhal ) (Coleoptera: Curculio nidae) (Bryan et al. 1993) is another good example of suc cessful biological control The alfalfa weevil, native to Europe, was originally detected in the U.S. in 1904. By 1970, the weevil had spread to all 48 contiguous states and become a serious pest of alfalfa ( Medicago sativa L., Fabaceae). I n 1957, the U SDA Agricultur al Research Service (USDA ARS) released parasitoid species belonging to four families (Eulophidae, Ichneumonidae, Mymaridae, and Braconidae) to reduce weevil populations to manageable levels in the eastern U.S. (Day 1981). The Risk Analysis P rocess for Entomophagous Biological Control Agents Risks Associated with the Importation and Release of Entomophagous Biological Control Agents The largest risk associated with the introduction and release of BCAs is their potential detrimental environmental effects on nontarget organisms (Louda et al. 2003). Unfortunately, once BCAs are successfully established, they are impossible to eradicate and their adverse impacts on nontarget organisms may persist indefinitely. For instance, the majority (83%) of parasitoids found attacking native caterpillars in Hawaii are BCAs intentionally released to control pest populations (Henneman & Memmot
27 2001). The refore, the main challenge for future biological control programs is to identify specific natural enemies of the targeted pests that are effective in their actions without having detrimental ecological consequences (Howarth 1983; Simberloff & Stiling 1996; Louda et al. 2003). Testing and verifying the existence and magnit ude of non ta rget impacts for entomophagous BCAs has become a significant concern of many biological control practitioners (Boyd & Hoddle 2007). De cisions to approve the importation and release of any new BCA must be considered carefully by appropriate authorities in the importing country, taking into account the risks of doing nothing or the risks associated with the BCA introduction, and the consequences of possible nontarget effects (Jayanth et al. 2003) including the likelihood that the agent will spread beyond national borders ( v an Lenteren et al. 2006). Several countries have developed new legislation and/ or have revised existing regulations to facilitate the introduction of new biological control organisms while minimizing environmental risks (COSAVE 1996; AQI S 1997; ERMA 1 997a; b). These regulations have mostl y been based on International Standards for Phytosanitary Measures ( ISPM # 2, 11 ) developed by the International Plant Protection Convention (IPPC) (De Nardo & Hopper 2004). Pest Risk Analysis The level of risk associated with the unintentional introduction of invasive species has been an ongoing concern that has been elevat ed to the international trade and environmental policy agendas (Andersen et al. 2004). Consequently, in 1995, the World Trade Organization (WTO) Agreement on the Application of Sanitary and Phytosanitary Measures (SPS Agreement) was ratified by most of the WTO country members (Ebbels 2003). Under the SPS A greement member countries agreed to base
28 their phytosanitary decisions on the results of a sciencebased assessment of potential risks (Andersen et al. 2004). In the area of plant health, this process was called pest risk analysis (PRA). In the U.S., t he task of protecting American agriculture and natural resources against the risks associated with the entry, establishment, or spread of plant pests is the responsibility of the U.S. NPPO the Plant Protection and Quarantine (PPQ) group within the Animal and Plant Health Inspection Service (APHIS) an agency within the USDA (APHIS 2007). The Plant Protection Act (PPA) of 2000 authorized the Secretary of Agriculture to delegate his/her plant protection authority to employees of USDA APHISPPQ. This federal entity has broad authority to develop and enforce phytosanitary measures that will prevent or delay the introduction and spread of plant pests ( PPA 2000 ). In the last 10 years, new and revised legislation and regulations have been implemented by the U.S. NPPO in response to the identification of new pathways for potential pest introductions resulting from trade globalization, diversification in transportation and increases in tourism (De Nardo & Hopper 2004). Risk can be defined as the combination of the likelihood of an adverse event and the magnitude of the consequences (Delfosse 2005). Pest risk analysis has three components. Several publications recognize different steps but essentially the key elements are: risk assessment which is the estimation of the likelihood and conse quences of an ad verse outcome; risk m anagement which identifies management options that will reduce or manage the adverse event; and r isk communication, which is a t wo way exchange of information about the undesirable event (Fisher et al. 1994). A PRA provides the techn ical justification for the application of phytosanitary measures.
29 In the area of biological control, a PRA examines the potential adverse effects that the introduction of BCAs can have and these include detrimental economic and/or environmental impacts (D elfosse 2005). It provides the scientific evidence to either allow or deny the importat ion and subsequent release of BCAs (Barratt & Moeed 2005). The PPA makes a specific distinction between plant pests and BCAs used in pest management programs. Under the PPA, the U.S. NPPO h as authority to regulate the importation and release of BCAs (PPA 2000). In addition, the National Environmental P olicy Act (NEPA) of 1973 requires the development and submission of an Environmental Assessment (EA) and Environmental Impact Statement (EIS) before approval to introduc e any organism with potential negative environmental impacts. An accepted and preferred alternative during the permitting process of entomophagous BCAs is the preparation and submission of an EA which provides information on the organism and its potential environmental significance in a succinct format (Kubasek & Silverman 2005). The EA document provides basic information on the positive and negative environmental impacts of the proposed action, including a management program in case of adverse consequences, and also suggests alternatives to the plan under consideration. The E A is compulsory under three circumstances: when the activity is federal ; when the federal activity is major ; and when the proposed activity will have a significant impact on the human env ironment (Kubasek & Silverman 2005). The E A is reviewed by an external group of experts selected by the agency required to file the E A The group of experts provides an analysis of potential environmental adverse effects of the proposed action. The revised document is then sent to the Council on Environmental Quality (CEQ) for further comments. Finally, in accordance with the
30 Administrative Procedure Acts rules on informal rule making, a draft is published in the Federal Register and public comments are submitted for a period of 90 days. When an EA is completed and determined that there are no significant impacts, no EIS needs to be filed (Kubas ek & Silverman 2005). In the U.S., c onsensus among regulators, decisionmakers, and biological control stakeholders on an acceptable and standardized r isk a ssessment framework for entomophagous BCAs has not yet been reached (De Nardo & Hopper 200 4 ; Boyd & Hoddle 2007). In the 1990s, scientists and governments worldwide identified a need for a harmoniz ed regulatory framework during the importation of BCAs in order to ensure a more effective plant and anim al protection system (Ebbels 2003). In Europe, an expert panel from the European and Mediterranean Plant Protection Organization (EPPO) published two guidelines on the safe use of biological control and developed a safe list for commercially used BCAs (Bigler et al. 2005). Independently, the European Uni on (EU) funded a research project entitled Evaluating Environmental Risks of Biological Control Introductions into Europe (ERBIC) and developed a document for the environmental risk assessment of exotic natural enemies in inundative biological control (v an Lenteren et al. 2003). In 1999, the Organization for Economic Cooperation and Development (OECD) developed guidance documents on appropriate regulations for the use of BCAs (Bigler et al. 2005). Recently, an EU policy support action document compared regulation procedures used by the EU and U.S. to develop a more efficient process for the EU (REBECA 2011). O ne of the priorities of the U.S. NPPO has been to improve their understanding and communication of the risks of the importation of
31 BCAs through t he development of a more transparent risk assessment process (Fisher et al. 1994). The SPS Agreement recognizes the IPPC as the standard setting body for phytosanitary measures to protect plant life and plant health. In response to an increasing demand fo r the use of BCAs across the globe, the IPPC developed the Code of Conduct for the Import and Release of Exotic Biological Control Agents as the third International Standard for Phytosanitary Measures ( ISPM # 3) (IPPC 1997) In 1995, t he standard was accepted by Food and Agriculture Organization (FAO) member countries. This standard provided a framework for safe importation and release of BCAs in the context of plant protection (Greathead 1997). In 2005, the scope of ISPM # 3 was expanded to include other beneficial organisms (e.g. sterile insects) and the title was changed to Guidelines for the Export, Shipment, Import and Release of Biological Control Agents and other Beneficial Organisms ( IPPC 2005). To conform to the SPS Agreeme nt the revised ISPM # 3 required that a NPPO be identified and made responsible for the implementation of phytosanitary measures. The revised ISPM # 3 refers to other phytosanitary standards that provide guidance in the development of PRAs ISPM # 2 ( Fra mework for Pest Risk Analysis) and ISPM # 11 (Pest Risk Analysis for Quarantine P ests I ncluding A nalysis of E nvironmental Risks and L iving M odified O rganisms) (IPPC 2004; IPPC 2007 ). The revised ISPM # 3 recommends that a dossier be developed for each agent and beneficial organism that includ es information on the organism pests targeted, potential human and animal health safety issues, and potential economic impact s of both the agent or beneficial organism and the
32 pest In the U.S. the evaluation of this dossier is a central component of the decision making process to allow or deny the importation of BCAs or beneficial organisms Permitting Process The enabling legislation that governs the importation and release of entomophagous BCAs in the U.S. is the PPA of 2000. To obtain approval for (1) importation of BCAs into containment facilities; (2) domestic movement of imported BCAs to other containment facilities; or (3) release of a BCA into the environment, applicants must complete a USDA APHISPPQ federal permit application form (PPQ 526) Furthermore, if the entomophagous BCA has not been previously released in the U.S., the appli cation must be accompanied by supplemental documentation which describes the justification for the proposed action, pr ovides information on the biology/ecology of the pest and the entomophagous BCA, the economic impacts and any potential detrimental environmental impacts as well as possible mitigation options (Hunt et al 2008 ). The dossier format is loosely based on the Guidelines for Petition for First Release of Non indigenous Entomophagous Biological Control Agents, Regional Standard for Phytosanitary Measures # 12 published by the NAPPO. The U.S. NPPO makes the final determination to approv e or deny a petition based on the information provided by the applicant during the permitting process Applicants who are employees of a federal agency, have received any federal funds and/or have employed any federal workers during the project must also write an EA as require d by NEPA (NEPA 197 0 ). Currently, t he U.S. has no standardized or published regulatory framework for importing and releasing entomophagous BCAs (Messing 2005) However, the regulatory framework for importation and release of non native phytophagous BCAs is
33 relatively well defined (Scoles et al. 2008). As regulations and requirements for importation and release of entomophagous BCAs are being developed, there has been an attempt by the U.S. NPPO to develop more transparent processes for entomophagous agents. Risk Communication Framework during the Importation of Entomophagous Biological Control Agents Risk Communication during Pest Risk Analysis International Standard for Phytosanitary Measures # 11 describes PRA as a process requiring three stages. Stage 1 is an initiation stage that involves identification of what triggers the PRA process. There are generally two initiation points for PRA the identification of a pathway or the identification of a pest that may qualify as a quarantine pest In stage 2 pest r isk a ssessment a risk assessment is conducted to evaluate the probability of introduction and potential economic and environmental consequences. Stage 3, r isk m anagement, requires the identification of mitigation optio ns to control/manage identified risks and some assessment of their effectiveness ( IPPC 2005 ). However, according to Fisher et al. (1994) a PRA should include: risk a ssessment that estimat es the likelihood of occurrence of a hazard and magnitude of the consequences; risk m anagement which addresses what can be done to mitigate the conseque nces of the adverse event; and risk c ommunication which involves twoway exchange of information concerning the likelihood and m agnitude of the hazard and the r is k m anagem ent measures to deal with the hazard. Risk c ommu nication is often neglected in PRA One of the reasons might be that risk communication is a non technical and subjective concept that is difficult to grasp for many agencies. Risk c ommunication also carries a stigma as it is often viewed as a
34 means to make stakeholders more rational (Jasanoff 1989; Goldman 1994). International Standard for Phytosanitary Measures # 11 contains only one sentence explaining the importance of communication of risks in the PR A process. Risk communication is an interactive exchange of information about a potential risk between individuals (Fischnoff 1990). Risk c ommunication can be described as a consensus structure that joins the interests and needs of both senders ( applican ts and researchers ) and recipient s ( governmental entities regulators, and decision makers). The main purpose of risk c ommunication is to provide individuals with enough information to enable them to make an informed decision ab out a potential risk ( Gibso n 1985; Fischnoff 1990; Gow & Otway 1990). In addition, risk c ommunication helps identify and explain the benefits gained by accepting a particular risk (Morgan et al. 2002). Mature risk communication is defined as more than providing adequate information. It is a conscious effort to develop a partnership between the senders and the receivers of information (Fischnoff 1995). Improving Risk Communication in Governmental Agencies Governmental agencies have long been concerned with communication methods used to more appropriately convey the risks associated with environmental issues (Chess et al. 1995). The U.S. NPPO has a long history of evaluating stakeholder satisfaction with its risk c ommunication efforts (Fisher & Chen 1996). It was one of the first agencies within the USDA to conduct a baseline survey of a wide range of stakeholders to examine how well they meet their customer needs during their risk communication activities (Fisher & Chen 1996). In 1990, the now defunct National Biological Control Institute (NBCI) was created within USDA APHISPPQ with the objective of promoting, facilitating, and providing
35 leadership in biological control and integrated pest management (APHIS 1996). From 1991 to 1995, NBCI gathered opinions on biological control regulations and related guidelines from a plethora of stakeholders (researchers from universities, integrated pest management working groups, professional society members, industry representatives, environmental groups, federal and state agricultural department officials). The Institute published its final report in 1996 highlighting areas in the current biological control regulatory system that needed improvement (APHIS 1996). One of those areas was customer service and communication activities during the regulatory process es associated with importation and release of phytophagous and entomophagous BCAs ( APHIS 1996). In 2006, an internal evaluation of USDA APHISPPQ permitting process for entomophagous BCAs considered that customer service was s till an activity that needed attention in order to improve BCA permitting activities in the U.S. (APHIS 2006). Approaches to Communicating Risk The one way model of communication includes a source that generates a message sent via a channel to a receiver ( Shannon 1948). Technical communication is the communication of scientific information. Risk communication is a subset of technical communication and is described as the communication of potential risks (Lundgren & McMakin 2004). In the process of r isk communication one must consider how the messages are sent and received, how conflicts and misunderstandings are managed, and how decisions are made (Lundgren & McMakin 2004). A few examples of methodologies used in r isk communication include the c onvergence communication approach, the m ental noise approach, and the mental m odel s approach. Each method examines risk communication processes based on how the audience perceives
36 risk (Lundgren & McMakin 2004). For instance, during the convergence communication approach, the values (culture, experience, background) of the audience must be taken into account as they may affect the risk communication procedure. A group within the U.S. Department of Defense uses the m ental n oise approach to communic ate with their stakeholders. This approach takes into consideration that peoples ability to process information might be altered by their feeling of being at risk (Blakeney 2002). The U.S Environmental Protection Agency used the m ental m odels approac h during the communication of radon informational programs to their stakeholders This approach allowed the integration of peoples understanding and view s on their current risk situation in the development of the risk message. Mental Models Approach wit hin Organizations The importance of understanding m ental m odels within an organization was first demonstrated in large corporations such as Royal Dutch/Shell. In the 1970s, the world oil business became more multicultural and a need for building consensus across the different management styles became important for oil compan ies A management consensus was only made possible by understanding the shared mental models of each cultural group (Sengue 1992). In order to build effective communication framework s across multicultural groups the knowledge gaps, general understanding, and misconceptions of each group first needed t o be identified ( Jungermann et al. 1988; Lave & Lave 1991; Bostrom et al. 1992; Maharik & Fischnoff 1992). Mental m odels are representations of the stakeholders assumptions based on personal perceptions. The assumptions can be simple generalizations or complex theories. Mental models help to anticipate and predict the outcome of an unknown event and usually impact the decision making process (Sengue 1992). The mental
37 m odels approach attempts to solve communication problems faced by risk specialists by requiring both a consideration of how the stakeholders intuitively think about the risks and which aspects of the sci entific literature actually matter to stakeholders (Morgan et al. 2002). The approach is based on a systematic analysis of the beliefs of decision makers and stakeholders and what specific information each group needs in order to make an informed decision (Morgan et al. 2002). Conditions for Using the Mental Models Approach Past studies ( Jungermann et al. 1988; Lundgren & McMakin 2004) have demonstrated that the m ental m odels approach is best used when the main purpose of risk communication is to modify behaviors. Additionally, the risk must be associated with conflicting opinions about how to manage the risk and the decisionmaking process should not be under time constraints. In this dissertation, the mental models approach was used to propose an improved risk communication framework for the permitting process of entomophagous BCAs as implemented in 2007 in the U.S. The analysis is discussed in Chapters # 3 (p. 49), # 4 (p.82), and # 5 (p. 104). The discussion below focuses on the other risk criteria us ed during risk assessment for importation and environmental release of entomophagous BCAs. Practical Application of Pest Risk Analysis into Biological Control Research Selecting a Successful Entomophagous Biological Control Agent Selection of an effective entomophagous BCA is a crucial step in the implementation of a biological control program. Various factors are used to determine the effectiveness of the BCA before its release into the environment. Biological parameters are used as technical justificati on for approval or denial of the introduction
38 of an entomophagous BCA. Host range, functional response, climate suitability, dispersal ability, high reproduction rate, and short life cycle, are just a few examples of critical biological parameters that ca n be used to examine or determine the future effectiveness of a particular BCA. Much of this same information is used to assess the risks associated with the importation of the agent. Biological characteristics In order to select a successful agent, the n atural enemy should display certain desirable attributes that relate to field performance. Previous studies demonstrated that abiotic factors, such as the presence of a source of food for energy have a positive impact on the establishment of introduced natural enemies (Boivin et al. 2006). The potential BCA should be host specific. In other words, the BCA should only feed on or parasitize the target pest species or closely related species that are also pests. This characteristic is theorized to reduce u nintended non target effects The natural enemy should also exhibit several key traits, especially a high reproductive capacity an effective attack rate, and good capacity for dispersal and searching (van Lenteren et al. 2006). Functional r esponse and numerical response Biotic factors such as the temporal or spatial availability of host s are important in the establishment of the BCA Functional response describes the behavioral response of an individual biological control agent to increasing host density (Holling 19 5 9). Results from these studies provide information on searching ability and handling time for BCAs. The concept is associated with the numerical response, which corresponds to the number of progeny produced as a function of host density (Hol ling 19 5 9)
39 Without a host density dependent numerical response, parasitoids are less likely to be able to reduce host populations (Holling 1959). Host suitability and preferences One of the key parameters used to determine potential nontarget effects by BCA s is the range of species that the BCA is able to attack. Therefore, host specificity testing has become the central issue in analyzing the risk of a potential BCA (DeNardo & Hopper 2004) While methods to determine host specificity of BCAs used to control weeds have been significantly improved (McEvoy 1996), this is not the case for BCAs used for the control o f insect pests (Barratt et al. 2000; Kuhlmann et al. 2006 ). Even though a focus on nontarget effects of non native enemies is considered th e key to safety associated with biological control programs (Haye et al. 2005), an effective and standardized host range testing strategy for entomophagous BCAs has not yet been developed (Duan & Messing 2000; Messing et al. 2006). The well accepted centr ifugal phylogenetic method (Table 11) developed by Wapshere (1974, 1989) is used to select host range test species of phytophagous BCAs. The method is based on phylogenetic taxonomic affinities of related plant taxa. Retrospective studies demonstrated o f established phytophagous BCAs have revealed that the host range of these insects is limited to phylogenetically related host plant taxa (Bernays 2000; Pemberton 2000; Kuhlmann et al. 2006). There is little or no evidence in the existing literature that this approach has failed (McFayden 1998) This method is complicated for entomophagous BCAs because of unreliable host lists (due to possibly greater number of host taxa attacked), difficulty of assessing behaviors influenced by habitat under laboratory conditions, difficulty in mass rearing nontarget test species, and possible host shift s by BCAs ( van Lenteren et al. 2 006; Boyd & Hoddle 2007). In addition, until the mid 19 80s
40 parasitoids and predators of plant pests were not subjected to l aboratory host range tests unless they were thought to be capable of attacking beneficial organisms and/or other important nontarget species (Ertle 1993). Pre release studies to determine the susceptibility of nontarget plants has been part of weed biological control for over 60 years (Barratt et al. 2000 ). T he prerelease host specificity testing for entomophagous BCAs is still one of the most difficult tasks tha t an applicant faces during the permitting process for entomophagous BCAs (Barratt et al. 19 98). The extremely varied nature of host parasitoid relationships and the large number of insect taxa precludes the establishment of a single prescriptive set of protocols for parasitoid host specificity testing (Barratt et al. 1998). However, a process similar to weed biological control using phylogenetic and ecological parameters for possible hosts and non target species might be applied. In addition, information on parasitoid host range from the country of origin can be indicative of the possible extent of the range of species which might be at risk in the area proposed for release (Barratt et al. 2000). A review of published methods used to establish host lists for entomophagous BCAs showed that phylogenetic criterion was an important component Bio logical, ecological, and socioeconomic information were also important considerations during the process. Several studies (Duan & Messing 1996; Duan et al. 1997; Barratt et al. 1997, 1998, 2000) used these criteria during the development of host test lis ts, and subsequent post release monitoring of inundative parasitoid releases showed few nontarget impacts on hosts from different feeding niches (Duan & Messing 1996; Duan et al. 1997). The results demonstrated that host morphological features were as im portant
41 as phylogenetic relationships for host selection to determine oviposition behaviors of parasitoids. Example of NonTarget Impacts: Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), a Spreading Pest in the U.S. The Argentine cactus moth, Cactoblastis cactorum often is referred to as an example for successful biological control of weeds (Moran & Zimmermann 1984). Prickly pear cactus, Opuntia sp. were introduced into Australia in 1788 as ornamental plants and again in 1830 as hosts of the cochineal dye producing scale insect, Dactylopius coccus Costa (Hemiptera: Dactylopiidae). Unfortunately, the Opuntia cacti became invasive, resulting in th e loss of millions of hectares of productive rangeland. In 1925, C. cactorum was introduced into Australia from its native Argentina to control the prickly pear. The cactus moth diet is restricted to the genus Opuntia but has a wide number of hosts in th is genus in its native geographi cal range (Moran & Zimmermann 1984). Gregarious C. cactorum larvae destroy Opuntia cacti by feeding internally i n the pads (or cladodes) (Dodd 1940). Within a few years, the introduction of C. cactorum into Australia resto red US $6 m illion worth of rangeland to agriculture at that time (Dodd 1940). Based on these encouraging results, C. cactorum was transferred from Australia into South Africa, Mauritius, and Hawaii to manage other nonnative Opuntia species that had becom e weeds in those countries (Moran & Zimmermann 1984). In 1957, C. cactorum was introduced into several Caribbean islands (Nevis, Montserrat and Antigua) to control native and nonnative Opuntia (Simmonds & Bennett 1966). The C. cactorum stock introduced into Australia and subsequently into other parts of the world was collected from Opuntia delaetiana (Weber) (now known as O. paraguayensis Schumann) and from an Opuntia species of the monacantha group (McFa y den 1985).
42 All introduced populations originated from this one introduction to Australia in 1925, which comprised about 3,000 eggs. Unfortunately, little thought was given to the potentially injurious environmental impacts if C. cactorum were to arrive in the U.S. (Stiling et al. 2004). The first record of C. cactorum in the U S was from Bahia Honda Key, Florida, in October 1989 (Dickel 1991). The pathway by which the moth first arrived in Florida is not known. The moth either spread naturally or was unintentionally transported from the Caribbean with imported Opuntia nursery stock (Pemberton 1995; Stiling 2002). In Florida, there are six native Opuntia spp. (Benson 1982). Three species are widespread: Opuntia stricta (Haw.) O. humifusa (Raf.) and O. pusilla (Haw.). Three a dditional species are considered rare in the Florida Keys: O. corallicola (S mall ) O. cubensis Britton & Rose and O. triacantha (Willd.). In addition, four naturalized species also are common in Florida: O. ficus indica (L.), O. monacantha (Mills.) O. leucotricha DC., and O. cochenillifera (L.) (Johnson & Stilling 1996, 1998). Cactoblastis cactorum spread rapidly and all Florida Opuntia spp. wer e attacked (Johnson & Stilling 1998). As many as three Opuntia spp. in the U.S. are used by people as food, animal fodder, and for dye production (Zimmermann et al. 2000). Concerns about environmental nontarget effe cts w ere initially greatest for cacti endemic to the Florida Keys, especially O corallicola (Johnson & Stilling 1996, 1998), one of the rarest plants in North America (Austin et al. 1998). At that time, the total worlds population of O corallicola consisted of 12 plants located on a property protected by t he Nature Conservancy at Little Torch Key (Gordon & Kubisiak 1998; Stiling et al. 2004). Ad ditional isolated populations of O. corallicola were subsequently found in South Florida (Hight et al. 2002).
43 Inundative Biological Control as a Pest Management Strategy for Cactoblastis cactorum Pemberton and Cordo (2001a, b) review ed available natural enemies and pathogens to control C. cactorum Several larval and pupal parasitoids were reported from South America including species from the families Braconidae, Chalcidae, Ichneumonidae and Tachinidae. In general, these natural enemies were f ound to parasitize the pest with a moderate success rate (<30%) For instance, the braconid wasp, Apanteles alexanderi Brethes, parasitized approximately 30% of the larvae under field conditions (Parker & Pinnell 1973). Trichogrammatid egg parasitoids al so were recorded attacking C. cactorum in Argentina ( Logarzo et al. 2009). Parasitized eggsticks have also been reported from Florida (Bennett & Habeck 1995). Egg parasitoids of the genus Trichogramma have been used successfully as inundative BCAs agains t a range of agricultural pests such as corn borers, sugarcane borers and cotton bollworm [ Helicoverpa zea (Boddie)] (Li 1994; van Lenteren 2000). Trichogramma spp. parasitize more than 200 insect species belonging to 70 families in 8 insect orders, espec ially Lepidoptera (Parra et al. 1987). For example, Trichogramma spp. have been successful at controlling internal feeding lepidopterans such as the European pine shoot borer Rhyacionia buoliana Denis & Schiffermller (Kogan et al. 1999), and the Europe an corn borer, Ostrinia nubilalis Hbner (Dahlsten & Mills 1999). Although Trichogramma wasps are the most widely used entomophagous group in biological control programs (Li 1994), Bennett and Habeck (1992) were the first to suggest the possibility of their use to manage C. cactorum
44 Assessing the Risk of Releasing a Non Specific Egg Parasitoid to Control Cactoblastis cactorum in North America Trichogramma species are facult ative gregari ous egg parasitoids (Rabinovich 1970), attacking eggs of a wide range of lepidopteran and other insect orders ( Clausen 1940; Pinto & Stouthamer 1994; Thomson & Stinner 1989). As such, inundative releases of species of Trichogramma to control C. cactorum might result in adverse impact s on rare butterfly species such as those present in the Florida Keys (Bennett & Habeck 1992). Furthermore, Trichogramma wasps also may negatively impact native cactophagous moths that also produce eggsticks such as Melitara prodenialis Walker (Lepidoptera: Pyralidae) Previous studies demonstrated that the efficacy of parasitoids as biological control agents wa s influenced by several factors including host acceptance, host age, and contact time between host and parasitoid. Therefore, evaluation of host range and other biological parameters that influences host selection constitutes the first step for understanding the potential risks associated with parasitoid introductions. Because of current stringent requirements governing the importation of nonnative entomophagous BCAs into the U.S., it was worthwhile to explore the potential us e of egg parasitoids already present in Florida and known to attack cactus moth in the field (Pemberton & Cordo 2001 a ). In Austral ia, T minutum Riley was reported to parasitize up to 32% of the cactus moth eggs in the field (Dodd 1940). Indeed, inundative releases of T. minutum were used against many pest insects (Li 1994), including the sugarcane borer Diatraea saccharalis F (Lepidoptera: Pyralidae) in Florida (Wilson 1941). A nother potential egg parasitoid species was T. pretiosum ( Riley ), a parasitoid observed to parasitize C. cactorum eggs in the Florida Keys (Bennett & Habeck 1995).
45 Trichogramma (Hymenoptera: Chalcidoid ea: Trichogrammatidae) Presently, the superfamily Chalcidoidea has about 22,000 named species belonging to 19 families (Noyes & Valentine 1989; Noyes 1998). Most species are less than 3mm in length, averaging 1.5mm with the smallest about 0.11mm ( Dicopo morpha echmepterygis Mockford Male Mymaridae) (Noyes & Valentine 1989; Noyes 1998) Individuals from the family Trichogrammatidae are 1.8 mm in length including the ovipositor. The family currently includes 83 genera and 839 species. Trichogrammatids a re primary solitary or gregarious endoparasitoids of Lepidoptera, Hemiptera, Coleoptera, Thysanoptera, Hymenoptera, Diptera, and Neuroptera eggs (Strand & Vinson 1984) Trichogramma pretiosum (Riley) Trichogramma pretiosum a North American species, is thought to be the most common Trichogramma species found in the Western Hemisphere (Pinto et al. 1986; Olkowski & Zhang 1990; Zucchi et al 2010). This species has been documented parasitizing the cotton leafworm [ Alabama argillace (Hbner) Noctuidae] the velvetbean caterpillar [ Anticarsia gemmatalis (Hbner), Noctuidae], sugarcane borers ( Diatraea spp. Pyralidae), Heliothis armigera (Hbner Noctuidae), the cabbage looper [ Trichoplusia ni (Hbner ), Noctuidae] the tomato pinworm [ Keiferia lycopersic ella (Walshingham), Gelechiidae], the Indian mealmoth [ Plodia interpunctella (Hbner), Pyralidae] and others (Li 1994). Hassan (1993) documented that T. pretiosum was released commercially in the U S to control pests in to cotton, corn and soybean. W hre r and Hassan (1993) showed that T. pretiosum had a high egglaying capacity (53.7 eggs/ female ) Monje et al. (1999) showed that female wasps preferred to parasitize the smaller Sitotroga cerealella (Olivier) (Lepidoptera: Gelechiidae) eggs over
46 the larger Diatraea rufescens Box (Lepidoptera: Pyralidae), and Diatraea saccharalis (F.) eggs. Trichogramma fuentesi Torre Trichogramma fuentesi has been recorded in several countries in South America (Argentina, Columbia, Mexico, Peru, and Venezuela) and seven states in the U.S. (Alabama, California, Florida, Louisiana, New Jersey, South Carolina and Texas) (Fry 1989; Pinto 1999). The primary hosts of T. fuentesi are species belonging to the families Noctuidae, such as Helicoverpa zea (Boddie) and Heliot his virescens (F.), and Pyralidae, such as Diatrea saccharalis (F.), Ephestia kuehniella Zeller and Ostrinia nubilalis (Hbner) ( Fry 1989; Wilson & Durant 1991; Pintureau et al. 1999; Querino & Zucchi 2003). Trichogramma fuentesi is widely used for biolog ical control of orchard pests
47 Table 21. Selection criteria of the centrifugal phylogenetic method for choosing test plants to determine host range of weed biological control agents (after Wapshere 1974, 1989). Plant Group Host Range 1 Host plants with similar genetic types (ecotypes/biotypes) 2 Plant species from same genus 3 Host plants from same tribe 4 Host plants pertaining to same subfamily 5 Host plants from same family 6 Host plants belonging to same order
48 Figure 21. Life stages of Cactoblastis cactorum : A Female laying eggs on a cactus spine, B Larvae feeding inside Opuntia cactus pad, C Male pupa (right) and female pupa (left), and cocoon (center). A B C L. photo von Richter photo H. Robertson photo I. Baez
49 CHAPTER 3 COMPARISON OF REGULATORY PROCEDURES FOR THE IMPORTATION AND RELEASE OF ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS IN EIGHT COUNTRIES HOW IS IT DONE? C oncerns in recent years about potentially undesirable environmental impacts of entomophagous biological control agents (BCAs) have generated a need for development of a harmoni z ed risk analysis process for importation and release of these agents Consequently there has been i nterest in developing harmonized technical and regulatory frameworks for plant protection specifically in the area of biological control at the national, regional and international level s. While a risk based process has been an integral part of the importation process for weed BCAs in the U.S. this approach has been a relatively new consideration for entomophagous BCAs. Risk can be defined as a combination of the probability of an adverse event and the magnitude of the consequences (Delfosse 2005). In 2007, t he U S ha d no standardized regulatory framework for the importation and release of non native entomophagous BCAs (Messing & Wright 2006; Hunt et al. 2008) The level of risk associated with the unintentional introduction of invasive species has been elevated onto the international trade and e nvironmental policy agendas (Andersen et al. 2004). In 1995, the World Trade Organization (WTO) Agreement on the a pplication of Sanitary and Phytosanitary Measures (SPS Agreement) was ratified by most of the WTO country members (Ebbels 2003). Under the S PS A greement the member countries agree to base their phytosanitary decisions on the results of a science based assessment of potential environmental and economic risk s ( IPPC 2005 ). The International Plant Protection Convention (IPPC) is recognized as the standard setting body for the application of SPS principles to plant health matters. In 1995, a s a
50 result of the growing concerns from the scientific community and the general public about the potential negative impacts of BCAs, the Code of Conduct f or the Import and Release of Exotic Biological Control Agents (International Standard for Phytosanitary Measures / ISPM # 3) was endorse d by the IPPC. In 2005, the document was revised to include other beneficial organisms ( IPPC 2005). Other relevant pest risk analysis (PRA) standards include International Standard for Phytosanitary Measures (ISPM) # 2 Framework for Pest Risk Analysis (IPPC 2007) and ISPM # 11 Pest Risk Analysis for Quarantine Pests including Analysis of Environmental Risks and Living M odified Organisms (IPPC 2004). In addition, at the regional level, the North American Plant Protection Organization (NAPPO) developed the Guidelines for Petition for First Release of Exotic Entomophagous Biological Control Agents (Regional Standard for Phytosanitary Measures / RSPM # 12) (NAPPO 2008) Several regional blocks and/or countries have developed new legislation or have revised existing regulations to facilitate the introduction of new biological control organisms while minimizing their potent ial environmental risks (COSAVE 1996; AQIS 1997; ERMA 1997a, b). However, there have been few comparative studies of regulatory systems in different countries and much of the information is scattered in less accessible conference proceedings or National P lant Protection Organization (NPPO) websites A comprehensive and timely comparative analysis will provide critical information that will guide further development of appropriate policies and legislation and development of practical and effective regulatory processes for entomophagous BCAs. This study used ISPM # 3 to identify key criteria that should be used during the decisionmaking process for entomophagous BCAs The objective was to develop a
51 summary of regulatory approaches used by different countr ies as these frameworks relate to the criteria outlined in ISPM # 3. It is anticipated that th e findings of this study might allow countries to compare different approaches to risk analysis and facilitate development of national processes that support inf ormed decisions regarding the screening and approval of entomophagous BCAs. Methodology Selection of Countries An initial list of 15 countries was selected for the comparative analysis based on their implementation of an operational regulatory system for the importation and release of entomophagous BCAs. However, because of limited available information and difficulty in language translation, several countries (China, France, Indonesia, Pakistan, Philippines, Russia, and South Africa) were eliminated from consideration. A subset of eight countries (U S Australia, Canada, U nited K ingdom India, New Zealand, Mexico, and Switzerland) was selected for detailed analysis based on 4 criteria: the ample amount of available information, wides pread geographic representation, level of gross domestic product (G DP ), and implementation of ISPM # 3 Guidelines for the Export, Shipment, Import and Release of Biological Control Agents and Other Beneficial Organisms (IPPC 2005) (Table 31) The GDP A gricultural Sector g ave the contribution to agriculture relative to the total G DP The GDP and G DP Agricultural Sector were used as an index to select developed countries Ratification of the SPS Agreement was not a selection criterion, although most countries in the final list have ratified the protocol.
52 Collation of Information Internet search engines (e.g. Google Google Scholar or Google Books ) as well as bibliographical databases (e.g. Agricola, CAB abstract s) were used to gather relevant information on the NPPO structure, legislation/regulations and current risk analysis pro cess implemented by the eight countries during their permitting process of entomophagous BCAs The key words risk, risk assessment, risk analysis, plant protection organization, biological control, importation of biological control agents, and release of biological control agents were used to identify rel evant information. S election of the key words was subjective. Reviews of NPPO configuration, PRA process, and permitting system for importation and release for entomophagous BCAs in the AsianPacific, North American, and European Union countries were also used as source s of data (Fasham & Trumper 2001; Mason et al. 2005; RE BECA 2006; FAO 2007; Loomans 2007; Hunt et al. 2008). Comparative Process T he status of regulatory biological control program frameworks, as implemented in 2007, were evaluated for each of the eight countr ies Comparisons were made between the NPPOs structures, PRA framework, permitting process for entomophagous BCAs, and information required from the petitioner during the importation and release of BCAs Selected decision making criteria used by each NPPO during the regulatory process for entomophagous BCA introduction w ere summarized. The ISPM # 3 was used to identify crucial themes that are of importance to the decision making process and these were: designation of responsible authority, law s and regulations, general principles (general acceptance of precaution) communication and reporting,
53 documentary responsibilities, reviewing process and consultation, and responsibilities of NPPO before, during and after release. Results Australia (AU) Designation of authority, laws and regulatory requireme nts In Australia, the Department of Agriculture, Fisheries and Forestry (DAFF) is responsible for the Australian Governments animal and plant biosecurity policy and the establishment of risk management measures (Figure 3 1) The Secretary of the Department of Agriculture is also the Director of Animal and Plant Quarantine under the Quarantine Act of 1908. The Australian NPPO consists of AQIS (Australian Quarantine and Inspection Service), Biosecurity Australia (BA) and the Office of the Chief Plant Prote ction Officer which are all within DAFF (FAO 2007) The Biosecurity g roup within the Department takes the lead in biosecurity and quarantine policy development and the establishment and implementation of risk management measures across the biosecurity con tinuum (DAFF 2007; Hunt et al. 2008). The importation and release of entomophagous BCAs is also regulated by the Department of the Environment and Heritage (DEH) under the Environment Protection and Biodiversity Conservation Act of 1999 (DEH 2007; Hunt et al. 2008). General acceptance of precaution Australia expresses its Appropriate Level of Protection (ALOP) in qualitative terms. The objective of the ALOP is to provide a high level of phytosanitary protection while reducing risk to a low level (FAO 200 7). This approach conforms to the WTOs Agreement on the application of SPS Agreement (FAO 2007).
54 Documentary responsibility of importer An initial application to import entomophagous BCAs into quarantine includes information on biology, ecology, and potential economic and environmental impacts of the agent and the target pest. In addition, a risk benefit analysis is included. A host species specificity test list and the methodology used for testing also need to be provided to DAFF for approval (DAFF 2007; Hunt et al. 2008). After completing the host specificity tests and before releasing an entomophagous BCA, a release package is submitted by the permit petitioner to AQIS (DAFF 2007) and separately to DEH (DEH 2007) (Figure 31). The dossier provides information on the origin, biology, and native range of the agent as well as related species to the agent. Methods used and results from laboratory evaluation of the agent are discussed by the petitioner, including a list of potential nontarget organi sms, results from host specificity testing, an evaluation of risk of non target impacts, as well as the appropriate testing methods and statistical analysis (IRA 2007; Hunt et al. 2008). The current economic and environmental status of the target species is also assessed by the petitioner (IRA 2007; Hunt et al. 2008). The document indicates whether and when the agent was approved for biological control in other locations, host records from foreign countries, and results from previous risk assessments or e nvironmental assessments of the BCA in other countries. The permit petitioner needs to indicate any possible conflicts of interest with existing biological control programs (IRA 2007; Hunt et al. 2008). Australian regulatory process for importation and release of entomophagous BCAs (Figure 3 1) The importation and release of an entomophagous BCA must be approved by DAFF and DEH. In addition, t he host specificity test list must be approved by the co-
55 operators which include the AQUIS within DAFF (DAFF AQIS), the DEH, the Commonwealth Scientific and Industrial Research Organization (CSIRO), and relevant state/territory government departments or research organizations. Cooperators have the power to veto any decision if their recommendations are not accepte d by the petitioner. Reviewing process/consultation P ublic participation is sought at different stage s of the approval process ; for example, stakeholders are consulted prior to the importation and the release of entomophagous BCAs (Figure 31). Responsibility of NPPO during and following release of BCA Release of entomophagous BCA is supervised by a federal quarantine entomologist working for DAFF. Post release evaluations are recommended but not enforced by either DAFF or DEH (Hunt et al. 2008). Canada (CAN) Designation of authority, laws and regulatory requirements The Canadian Food Inspection Agencys (CFIAs) Plant Health Division (PHD) fulfills the obligations as Canadas NPPO CFIA PHD is responsible for pest risk assessment and management in Canada. Regulation of entomophagous BCAs was initially implemented in the late 1990s by the CFIA through the authority of the Canadian Plant Protection Act (PPA) of 1990 (DJC 2005). Pest risk assessments are developed and evaluated by the CFIAs Pla nt Health Risk Assessment Unit, within the Science Division.
56 General acceptance of precaution Pest risk assessment is a science based procedure based on information provided and results from host specificity testing. An ALOP approach is used during risk assessment (Mason et al. 2005). Documentary responsibility of importer Petitions for import for research purposes include information on the BCA (scientific name, common name, origin, etc), the proposed action, and a management plan to prevent the spread of target pest. To obtain release approval, the permit petitioner needs to submit additional information on both the BCA and target pest biology, and results from evaluations of host specificity testing. In addition, the possible environmental and economic impacts of the proposed action should be covered in the petition. The information should conform to the NAPPO standard requirements for entomophagous BCAs as described in RSPM # 12 (NAPPO 2008). In order to obtain a permit for release, the permit peti tioner needs to provide a detailed description of the agent, information on the release locations, means of transportation used, and reason for release. Canadian regulatory process for importation and release of entomophagous BCAs (Figure 3 2) Information submitted by permit petitioner is reviewed by CFIA PHD for approval for importation and movement of the BCA to a federal quarantine facility for research purposes (CFIA 2006; Hunt et al. 2008). For release approval, the permit petitioner must take into ac count recommendations from the Ontario Plant Laboratory Quarantine Entomology Laboratory ( OPL QEL ). Once the requirements from the OPLQEL are met, the petition is sent to the Biological Control Review Committee (BCRC) for review.
57 Based on recommendations from the BCRC after evaluating the application, the OPLQEL forwards their recommendation whether or not to approve the release of the proposed entomophagous BCA to the Director of the CFIA PHD. The Director of the CFIA PHD will either approve or deny t he release of the BCA or request that more research be conducted (CFIA 2006; Hunt et al. 2008). Reviewing process/consultation Public opinions are not solicited in Canada during the permitting process of entomophagous BCAs (Hunt et al. 2008). Prior to issuance of a permit for release purposes, CFIA PHD forwards petitions to the OPL QEL. The OPLQEL verifies that the information provided by the permit applicant follows NAPPO standards. Completed petitio ns are also sent to the BCRC for external review. The BCRC is composed of taxonomists, ecologists, entomologists, botanists, federal and provincial scientists, extension specialists, university researchers, and Environment Canada and Pest Management Regulatory Agency officials. Petitions are also sent t o the Mexican NPPO and the U.S. NPPO for comments (Figure 32) (CFIA 2006; Hunt et al. 2008). Responsibility of NPPO during and following release of BCA Although post release evaluations are not enforced by the Canadian NPPO ; the applicant must develop a plan of action to assess economic and environmental impacts. European and Mediterranean Region The European and Mediterranean Plant Protection Organization (EPPO) is an intergovernmental organization responsible for European cooperation in plant protect ion in the European and Mediterranean region. The organization currently has 50 member countries including Switzerland and the United Kingdom (EPPO 2006) The EPPO is administered by its Executive Committee (seven Governments elected on a rotational
58 basi s, meeting twice a year), under the control of its Council (representatives of all member governments, meeting once a year) headed by a Chairman and a ViceChairman, which are elected individuals. The technical work for the organization is done by Panels of Experts, under the supervision of Working Parties. Experts are nominated by their respective NPPOs (EPPO 2006) Most of the European countries have ratified the Convention on Biological Diversity (CBD) which stipulates that the introduction of nonnat ive species should be under regulatory control (REBECA 2006). In addition, applicants might need a permit for environmental release as stipulated by the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). However, bot h of these directives are directed to preserve natural habitats and native flora and fauna (Loomans 2007). The organization has regulations and laws to protect natural habitats and indigenous flora and fauna, but the organization does not have specific laws for the regulation of entomophagous BCAs (EPPO 2006; REBECA 2006) Each European country is responsible for developing the appropriate regulations and methodologies for PRA (REBECA 2006) In 1997, in an effort to provide guidance for safe use of BCAs between the European Union (EU) countries, EPPO and CABI Bioscience organized a workshop on safety and efficacy of BCAs (Bigler et al. 2005). As a result, guidance documents for the safe use o f BCAs for research purposes were developed: First I mportation of E xotic BCAs for R esearch under C ontained C onditions PM 6/1(1) (EPPO 1999) and Import and R elease of N onI ndigenous BCAs PM 6/2(1) (EPPO 2001a), along with a guidance document focusing ma inly on the PRA process (EPPO 200 1 b ). In parallel, the EPPO funded the Evaluating Environmental Risks of Biological Control Introductions into
59 Europe (ERBIC) project. The outcome was the development of a document with detailed criteria for risk assessm ent of BCAs to assist regulators in assessing environmental risks associated with their use (van Lenteren et al. 2003). In 1999, the Organization for Economic CoOperation and Development (OECD) was established. The OECD member countries worked to devel op a harmonized approach for regulation of BCAs. The initiative resulted in the development of a guidance document stipulating the information requirements during permit submission of BCAs (OECD 2004). However, the EU felt that information requirements proposed by the OECD guidance document would hinder the existing regulatory system for BCAs. Consequently, a commission of the International Organization for Biological Control/Plants West Paleartic Regional Section (IOBC/WPRS) produced a document that provided guidance for a streamline regulatory process for BCAs (Bigler et al. 2005). Recently, an international initiative was created to harmonize the regulatory systems for BCAs between U.S. and EU countries; the REBECA (Regulations for Biological Control Agents) project. One of the objectives of REBECA was to review regulations of BCAs and provide recommendations for an implementation for a more harmonized and efficient regulatory process for BCAs (REBECA 2011). The analysis for each risk assessment crit eria as outlined in ISPM # 3 will only be addressed for Switzerland and the United Kingdom. Switzerland (SW) Designation of authority, laws and regulatory requirements Switzerland has developed an operational regulatory system for importation and releas e of BCAs (REBECA 2006). The Federal Law for the Protection of the Environment regulates the use of entomophagous BCAs under the Ordinance on Plant
60 Protection implemented by the Federal Office for the Environment. In addition, specific ordinances are use d to assist in containment procedures of entomophagous BCAs : Ordinance on the Contained use of Organisms ( 1999) and for the environmental release Ordinance on the Release of Organisms into the Environment ( 1999) (Loomans 2007) General acceptance of precaution There is no standardized PRA process for the importation and/or release of entomophagous BCAs (REBECA 2006). The application process is structured according to procedures already in place for plant protection (REBECA 2006). Documentary responsi bility of importer The applicant must provide information as recommended by the guidance document developed by OECD (OECD 2004) on information requirements for regulation of invertebrates as biological agents (Loomans 2007). The documents require informat ion on the identity, origin, source, distribution, biology, native host range, natural enemies, and potential economic of the BCA. In addition, an evaluation of the human and animal health impacts of releasing the BCAs should be included. Specific environmental criteria, such as information on the establishment in the wild, host specificity data and information on nontarget effects are requested (Loomans 2007). Swiss regulatory process for importation and release of entomophagous BCAs The applicant mus t first submit an application to register the entomophagous BCA to the Federal Office of Environment. After registration, the applicant needs to apply for a permit for importation and to submit a dossier containing information as recommended by the OECD g uidance document (OECD 2004). Upon approval for importation,
61 decisions to grant approval for environmental release will be based on the quality and quantity of information provided by the applicant (Loomans 2007). Reviewing process/consultation No public hearings or consultations are solicited during the decisionmaking process. The Federal Office of Environment reviews information provided by the permit petitioner and makes final decision. Responsibility of NPPO during and following release of BCA A p ost release evaluation plan is not required by the SW NPPO. United Kingdom (UK) Designation of authority, laws and regulatory requirements To some extent, the UK has an operational regulatory process for BCAs (REBECA 2006). The Ministry of Agriculture Plant Health Division under the Great Britain Plant Health Act (1967) is responsible for all plant health matters. There is no specific legislation regarding the import and release of nonnative invertebrate BCA s for the purpose of biological control (Loomans 2007) General acceptance of precaution I n 1998 with the assistance of the Cabinet Office, the Prime Minister introduced a Regulatory Impact Assessment (RIA) procedure. All regulatory proposals must contain a RIA which is a n organized document that of fers a cost benefit analysis, risk assessment and compilation of potential stakeholders that might be impacted by the proposed action. In addition, a review of alternative nonregulatory options needs to be included in the RIA Risk communication is an important component of the UK RIA.
62 Documentary responsibility of importer The application process is structured according to procedures already in place for nature conservation. The UK does not have a specific form for the approval of importation or rel ease of entomophagous BCAs. However, it is recommended that applications follow the format as recommended in the Department for Environment, Food and Rural Affairs (DEFRA) website (DEFRA 2000). A n extensive amount of information is given on the DEFRA website on data requirements to support applications for permit s to release entomophagous BCAs Information on the origin, biology, source, and native host range of the BCA is required. The k ey requirement is information about the establishment potential of the BCA in the UK. Environmental assessments including evaluation of previous use and host specificity testing should be discussed (DEFRA 2000). For a nonnative species, DEFRA requires data to be generated, when not already available, in order to properly assess the survival in the environment (Loomans 2007) United Kingdom regulatory process for importation and release of entomophagous BCAs A permit must be obtained before importation or release of entomophagous BCAs. The application is reviewed by the competent authority and decisions are made based on the information provided by the applicant and the recommendations developed by the National Advisory Committee. Reviewing process/consultation The National Advisory Committee is comprised of experts from various disciplines. The Committee is consulted during the decisionmaking process (Loomans 2007). No public hearings or consultations ar e solicited during the decisionmaking process.
63 Responsibility of NPPO during and following release of BCA A post release evaluation plan is not required by the UK NPPO (DEFRA 2000). India (IN) Designation of authority, laws and regulatory requirements The D irectorates of Plant Protection Quarantine & Storage (PPQS) established under the Indian Department of Agriculture & Cooperation of Ministry of Agriculture have been given the responsibility of implementing the regulations relating to the import ation of BCAs in India. Their importation is regulated by the Plant Quarantine Order (2003) issued under the Destructive Insects and Pests Act (1914) and amendments issued there under (PPQS 2006) Importation of entomophagous BCAs requires a permit issued by t he Plant Protection Adviser. General acceptance of precaution The PPQS follows pest risk assessment approach as stated in ISPM # 11 (IPPC 2007) in relation to environmental risks as covering environmental aspects related to the use of entomophagous BCAs (PPQS 2006). Documentary responsibility of importer Prior to containment in a federal quarantine facility, the absence of contamination of the entomophagous BCA needs to be confirmed by a certificate issued by the NPPO of the country of origin (PPQS 2006). Prior to the first importation into India, information on the origin, distribution, biology, host specificity, and potential non target impacts of the entomophagous BCA should be submitted (PPQS 2006). In addition, the following information on the target pests should be provided to the Plant Protection Adviser: origin, biology, ecology, economic and environmental impacts, possible benefits and
64 conflicting interests surrounding its use, and known natural enemies, antagonists and competitors already present or used (PPQS 2006). Indian regulatory process for importation and release of entomophagous BCAs For importation approval, the permit petitioner must submit PQ 12 form to the Plant Protection Adviser at least two months prior to proposed action. The application is reviewed by a Technical Committee established under the chairmanship of the Department of Agriculture and Cooperation (PPQS 2006). Upon approval for importation, the PPQS might grant permission for carrying out experimental studies under confi nement in isolated fields. After receiving results from field evaluation and after consultation with Technical Committee, PPQS will approve or deny environmental release of the BCA (PPQS 2006). Reviewing process/consultation A Technical Committee compos ed of representatives from the Directorates of Plant Protection Quarantine & Storage, Indian Council of Agriculture Research, Project Directorate of Biological Control, National Centre of Integrated Pest Management, Forest Research Institute, National Bure au of Plant Genetic Resources, Division of Entomology, Nematology, and Plant Pathology from the Indian Agriculture Research Institute review applications and develop recommendations prior to approval for importation (PPQS 2006). There is no public notification or solicitation during the decisionmaking process. Responsibility of NPPO during and following release of BCA The PPQS ensures that monitoring of release of entomophagous BCAs takes place (PPQS 2006).
65 Mexico (MX) Designation of authority, laws and re gulatory requirements Under the Secretaria de Agricultura, Ganadelia, Desarollo Rural, Pesca y Alimentacion (SAGARPA) (Ministry of Agriculture, Livestock, Rural Development, Fisheries and Food), the Direccion General de Sanidad Vegetal is responsible for permitting the importation and release of nonnative entomophagous BCAs in Mexico. The importation and/or release of entomophagous agents is regulated through the Plant Health Act, articles 101 and 102 (1980) (Mason et al. 2005) General acceptance of precaution Risk assessment is based on concepts described in ISPM # 2 and ISPM # 11 (IPPC 2004, 2007). Documentary responsibility of importer The permit petitioner must provide information according to RSPM # 12 on the origin, distribution, biology and ecology, host specificity, natural enemies of the BCA, and results from previous use of the agent (Mason et al. 2005). In addition, evaluation of the economic and environmental impacts and general information (origin, distribution, biology, and ecology) of the target pests should be provided. Potential benefits and any conflict of interest associated with its use should be included in the document (NAPPO 2008). Mexican regulatory process for importation and release for entomophagous BCAs (Figure 3 3) The Direccion General de Sanidad Vegetal i n collaboration with the Centro National de Referencia de Control Biologico (National Center of Biological Control
66 Reference) reviews import application s and recommendations are given to the Plant Health Director o f the Department of Agriculture. Reviewing process/consultation If needed, additional consultations with the Consejo Nacional Consultivo Fitosanitario (National Consultative Phytosanitary Advisory Group) are made prior to environmental release. The National Consultative Advisory Group is compos ed of professionals from academics, research, and government. There is no public notification or solicitation during the decisionmaking process. Responsibility of NPPO during and following release of B CA Conforming to RSPM # 12, a prerelease plan must be developed for monitoring of nontarget impacts (NAPPO 2008). However, the plan is not enforced by the MX NPPO. New Zealand (NZ) Designation of authority, laws and regulatory requirements In New Zealan d, t he Environmental Risk Management Authority (ERMA) is responsible for reaching decisions concerning necessary phytosanitary issue s. The introduction of entomophagous BCAs falls under the Hazardous Substances and New Organism (HSNO) Act of 1998 (ERMA 20 06) The ERMA is comprised of three entities: the decisionmaking A uthority the A gency tee. The Authority is composed of experts appointed by the Minister of Environment. The Aut hority functions like a quasi judicial entity with court like procedures. The Agency provides guidance to the applicant and advices the ERMA on matters relevant to nati (Hunt et al. 2008)
67 B iological control agents are also regulated under the Biosecurity Act of 1993 by the Ministry of Agriculture and Forestry (MAF) which is in charge of New Zealand Import Health Standards (IHS) that prevent unwanted introduction of organisms (Hunt et al. 2008) General acceptance of precaution The New Zealand approach to assessing the risks of introduction of a BCA is based on a full ecological risk, cost and benefit analysis (ERMA 2006; Hunt et al. 2008). The HSNO A ct requires the need for caution where there is scientific and technical uncertainty (Sheppard et al. 2003). Documentary responsibility of importer For importation approval, the applicant must provide information on the proposed action, identity, biology, and ecology of the BCA. Containment specifications (physical and operational) must also be included. An assessment of the risks, costs and benefits of importing the BCA must be developed (Hunt et al. 2008). For approval of full or conditional release o f the BCA, comprehensive environmental risk, cost, and benefit assessments must be developed. Guidelines on processes to conduct analysis are based on the Information factsheet Estimating the Beneficial Effects of Biocontrol Agents and A Technical Guid e to Identifying Assessing and Evaluating Risks, Costs and Benefits available on the ERMA website (ERMA 2006). One important element of the risk evaluation is the assessment of host range (Hunt et al. 2008). New Zealand regulatory process for importat ion and release of entomophagous BCAs (Figure 3 4) For importation approval of an entomophagous BCA, the applicant must submit a permit application to ERMA for evaluation. At this stage, the Authority can decide
68 whether notification of the public of the proposed action is necessary. The application must also satisfy requirements from the MAFs Import Health Standards For release of the BCA, the applicant must first get in contact with the ERMA Agency staff for proper guidance during the process. The ER MA Agency staff evaluates the application which is then accessible for comments by the Minister, and other interested parties. Based on ERMA Agency staff develops a report for the decisionmaking Authority. The Authority makes the decision to deny the BCA for release or approve full or conditional release. Under conditional release the BCA can only be released under specific conditions. Reviewing process/consultation An important aspect of HSNO Act is the Publics Right to Know section which provides the provision for public notifications and hearings of the proposed action. Public notifications include summary statements in daily newspapers, in government newslet ters, or on the ERMA website. The public has an opportunity to comment on proposed action prior to importation and release of BCAs. In addition, the petitioner has an opportunity to set up a public hearing to provide any additional relevant information b efore a final decision is made (Hunt et al. 2008). Responsibility of NPPO during and following release of BCA Although post release evaluations are not enforced; the applicant must develop a plan of action to assess economic and environmental impacts. U nited States (U.S.) Designation of authority and law/regulatory requirements In the U.S., there is no well defined regulatory process for importing and releasing entomophagous BCAs (Messing 2005). The Plant Protection Act ( PPA 2000) gives
69 broad jurisdicti on to the U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Plant Protection and Quarantine ( USDA APHISPPQ ) to regulate the importation, interstate movement and release of entomophagous BCAs. Permit conditions are based on regul ations as stated in 7 CFR section 330 Parts 200212 (CFR 2001). Many of the regulations used today are the ones developed under the previous Federal Plant Pest Act of 1957 (Hunt et al. 2008). General acceptance of precaution Permits are issued only when sufficient safeguards are in place and BCAs do not represent or have an unacceptable level of risk (PPA 2000). Regulations for entomophagous BCAs became more stringent since the terrorist attacks of 2001. A Guilty until proven innocent approach is tho ught to be used during the decisionmaking process for importation and release of entomophagous BCAs ( Ruesink et al. 1995; Simberloff 1996, 2005) Documentary responsibility of importer For importation approval, along with the permit form (PPQ 526) gene ral information (scientific name, host range, and geographic origin) on the entomophagous BCA must be provided. Containment specifications of an imported BCA into an USDA APHISPPQ quarantine facility are also included. For field release approval, the applicant must submit a dossier with information that follows the format provided by RSPM # 12 (NAPPO 2008) (Figure 3 5). U.S. regulatory process for importation and release of entomophagous BCAs (Figure 3 5) For the importation and release of entomophagous BCAs a permit (PPQ 526) administered by USDA APHISPPQ must be completed and submitted seeking
70 approval to bring the BCA into a federal quarantine facility. If the applicant wanted to move the BCA to another quarantine facility, a separate PPQ 526 form must be submitted and approved by USDA APHISPPQ. An additional PPQ 526 application for field release approval is required from the applicant. At this stage, a dossier conforming to RSPM # 12 Guidelines for Petition for First Release of Exotic Entomophagous Biological Control Agents (NAPPO 2008) must accompany the PPQ 526 application. Federal employees who have received any federal funds for the project must submit an Environmental Assessment under the National Environmental Policy Act of 1972. If USDA APHISPPQ considers that the risk poses by the proposed action is high, an Environmental Impact Statement (E IS) is required. After consultation with Canadian and Mexican counterparts from NAPPO, USDA APHISPPQ decides whether the proposed action has possible nontarget impacts on endangered and threatened species. If potential for negative impacts is verified, USDA APHISPPQ must consult with the U.S. Fish and Wildlife Service (FWS). Under the Endangered Species Act (ESA) of 1973, submission of a BA will be required from the applicant. If the USDA APHISPPQ considers that the release of the entomophagous BCA will cause no deleterious impact, a determination of no jurisdiction is made and the BCA can be field released according to the state laws. Reviewing process/consultation During the decisionmaking process, representatives from FWS an d NAPPO are consulted. In 2007, there was no public notification or solicitation of stakeholder input during the decisionmaking process.
71 Responsibility of NPPO during and following release of BCA As indicated in RSPM # 12, a prerelease plan must be developed for monitoring of non target impacts (NAPPO 2008). Discussion A ll eight countries outlined above were found to have regulations for the importation and release of entomophagous BCAs implemen ted to a certain degree. However, the analysis showed that regulatory procedures were not harmonized across countries. Two countries (SW, UK) did not have a specific risk assessment approach or regulations for the approval of importation and release of e ntomophagous BCAs (Table 3 2) During the decisionmaking process, for all eight countries, the applicants were required to submit a dossier with information on the identity, biology, ecology, native host range, and distribution of the proposed entomophag ous BCAs. Different guidance documents and formats were used by countries during the process. However, all the guidance documents required an evaluation of potential environmental impacts (Table 3 3). For all the countries, evaluation of possible negati ve environmental impacts required an analysis of the host range and results from host specificity tests (Table 33). Comparative analysis of regulatory procedures of the eight countries demonstrated that six of them (AU, CAN, IN, MX, NZ, UK) used a form o f pa rticipatory/collaborativebased r isk analysis process during the decision making process (Table 34) Within the PRA process, six countries (AU, CAN, IN, MX, NZ, UK) had integrated subject matter expert consultations (Table 35). These experts have d ifferent background s and include government employees to environmental groups. In addition, two countries ( AU, NZ ) solicited public comments and one country (NZ) considered public hearings necessary prior to the importation and/or release of entomophagous BCA s (Table 35) Most of
72 the NPPOs (AU, CAN, IN, MX, NZ, US) required a post release monitoring plan but only IN enforced the completion of the plan (Table 36). This comparative analysis provided information on how to improve and implement a workable participatory/collaborative based procedure within the existing permitting process for entomophagous BCAs in the U.S Specific recommendations to improve the U.S. process include the use of a group of experts to provide recommendations to USDA APHISPPQ d uring the decisionmaking process. In addition, the public should have the opportunity to comment on the proposed action prior to environmental release of the entomophagous BCA. Nevertheless, difficulties in implementing these suggestions in the U.S. inc lude the Guilty until proven innocent approach used during risk assessment and the consequent stringent regulations applied to the importation and release of entomophagous BCAs. In addition, the fear of possible litigations might prevent the implementat ion of a more transparent regulatory process. Chapters # 4 and # 5 in this dissertation describe how the regulatory system for permitting entomophagous BCAs, as used in 2007, could be modified to improve the decisionmaking process.
73 Table 31. C riteria used in countries examined in the pest risk analysis comparative study (CIA 2011). Countries IPPC Membership WTO SPS Agreement Implementation Conformed to ISPM # 3 PRA G D P (million $US) GDP Agricultural Sector (%) World 61,963,429 6.0 Australia X 1995 X X 1,219,722 4.1 Canada X 1995 X X 1,563,664 2.3 European Union X 1995 16,106,896 1.9 India X 1995 X X 1,430,020 17.0 Mexico X 1995 X X 1,004,042 4.3 New Zealand X 19 9 5 X X 138,003 4.6 Switzerland X 1995 522,435 8.4 UK X 1995 2,258,565 1.2 US X 1995 X X 14,624,184 1.2
74 Table 32. Comparison of general acceptance of precaution during pest risk analysis and decisionmaking during permitting for entomophagous biological control agents in 8 countries (Fasham & Trumper 2001; Mason et al. 2005; REBECA 2006; FAO 2007; Loomans 2007; Hunt et al. 2008). General Principles AU CAN SW UK IN MX NZ US General Acceptance of Precaution Appropriate Level of Protection approach reduces risk to a very low but not to zero X X X Full ecological risk, cost and benefit analysis X X Guilty until Proven Innocent approach X X X Specific Time Frame X X X X X Process in Place for Decision Reversal X X X X
75 Table 33. Comparison of documentary responsibilities of importer, prior to import, for pest risk analysis and decisionmaking during permitting for entomophagous biological control agents in 8 countries (Fasham & Trumper 2001; Mason et al. 2005; REBECA 2006; FAO 200 7; Loomans 2007; Hunt et al. 2008). Documentation Requirements AU CAN SW UK IN MX NZ US Documentary Requirements Related to the Target Organism Accurate identification of the target organism(s) generally at the species level X X X X X X X X Its known biology and ecology X X X X X X X X Its economic importance and environmental impact X X X X X X X X Possible benefits and any conflicting interests surrounding its use X X X X Known natural enemies, antagonists and other BCAs or competitors of the target pest X X X X X Documentary Requirements Related to the BCA Sufficient characterization of the BCA for accurate identification to the species level at minimum X X X X X X X X Voucher specimens deposited in recognized specimens X X X Key published X Summary of all available information on its origin, world, distribution, biology, natural enemies, hyperparasites and impact on its area of distribution X X X X X X X Information on host specificity and any potential hazards posed to non target X X X X X X X X Approval of host specificity list required by Federal Agency X Approval of host specificity not required but recommended X X
76 Table 34. Comparison of communication and reporting processes during pest risk analysis and decisionmaking during importation and release of entomophagous biological control agents in 8 countries (Fasham & Trumper 2001; Mason et al. 2005; REBECA 2006; FAO 2007; Loomans 2007; Hunt et al. 2008). Communication and Reporting AU CAN SW UK IN MX NZ US Clear Risk Assessment Guidelines and Policies Required information for applications is available on website X X X X X Risk assessment criteria publicly available online X X Public Notifications Notification of proposed release X X Applications available online before evaluation Risk assessment posted on public website Risk assessment summary posted on government website X X Risk assessment summary published in daily newspaper X Risk assessment summary published in government newsletter X X Release of risk assessment published in Federal Register X Public notifications of approved release Community informed about issues relating to safety X X
77 Table 35. Comparison of reviewing and consultation processes for pest risk analysis and decisionmaking during permitting process for entomophagous biological control agents in 8 countries (Fasham & Trumper 2001; Mason et al. 2005; REBECA 2006; FAO 2007; Loomans 2007; Hunt et al. 2008). Reviews and Consultation AU CAN SW UK IN MX NZ US Public Participation Solicit public comments in risk decision process prior to importation X Solicit public comments in risk decision process prior to release X X Formal procedures in place for hearings during decision process X Approval process includes public comment periods X X Use of Secondary Sources Use of risk assessments from foreign countries X Use data or results from previously submitted risk assessments X Use of Experts Consultation with scientific experts X X X X X X Consultation with members of regulatory body X X X X X X
78 Table 36. Comparison of responsibilities of the National Plant Protection Organization (NPPO) before, during and following release of biological control agent in 8 countries (Fasham & Trumper 2001; Mason et al. 2005; REBECA 2006; FAO 2007; Loomans 2007; Hunt et al. 2008). Responsibility AU CAN SW UK IN MX NZ US Release Release supervised by quarantine entomologist X Documentation Documentation on measures undertaken to ensure levels of contamination acceptable to the importing NPPO X X X Monitoring and Evaluation Post release monitoring enforced X Post release monitoring required but not enforced X X X X X
79 Figure 31. Australian permitting process for entomophagous biological control agents (Based on Hunt et al. 2008). Petitioner : Submit application for import into quarantine facility DAFF AQIS: Review and evaluate petition No DAFF BA Consultation Submit comments and recommendations Approval to import granted? Petition denied Yes Petitioner : Submit application for acceptance of list species for specificity testing Letter of approval for release Public notification (10days) for comments DEH: Review and evaluate petition DAFF BA : Review and evaluate list species DEH Consulta tion Submit comments and recommendations DAFF AQIS Consultation Submit comments and recommendations Approval to import granted? No Yes Petitioner : Submit application for release DEH: Review and evaluate petition DAFF AQIS: Review and evaluate petition Public notification (20days) for comments List species tabled in federal Parliament List species publishes in Government gazette Application denied
80 Figure 32. Canadian permitting process for entomophagous biological control agents (Based on Hunt et al. 2008). Petitioner: Submit application for importation accompanied with dossier as described in RSPM#12 Gu idelines for Petition for First Release of Exotic Entomophagous Biological Agents (NAPPO 2008) Review by Regulatory Entomologist of Quarantine Entomology Lab CFIA Import Service Revise petition for permit CFIA Plant Health Division Director Review petitions and recommendations Should the BCA be released? External Review U SDA -APHIS-PPQ (US) Sanidad Vegetal (Mexico) Yes No Issuance Letter Rejection Letter Biological Control Committee Review
81 Figure 33. Mexican permitting process for entomophagous biological control agents (Based on Mason et al. 2005). Petitioner : Submit application for importation of entomophagous BCAs Certificate of purity from phytosanitary authority of exporting country Certificate of origin from phytosanitary authority of exporting country General Director of Plant Health of Department of Agriculture (Sanidad Vegetal ) No National Center of Biological Control Reference Is the BCA approved for importation? Application denied Yes Petitioner: Application and information for release as recommended by NAPPO Letter of Approval for Release National Consultative Phytosanitary Advisory Group General Director of Plant Health of Department of Agriculture (Sanidad Vegetal ) Is the BCA approved for release ? Yes
82 Figure 34. New Zealand permitting process for entomophagous biological control agents (Based on Hunt et al. 2008). Petitioner : Prepare application for importation of entomophagous BCAs ERMA Review application No Public Consultation Minister and other appropriate parties/Public Notification Application denied ERMA Evaluate application and information Letter of Approval for Release ERMA Decision Making Committee Reviewed application and reports Public Hearing Is the BCA approved for release ? Yes ERMA staff consultation Petitioner : Submit application for importation of entomophagous BCAs Maori Group consultation consultation Experts Group consultation Appeal Process
83 Figure 35. U.S. permitting process for entomophagous biological control agents as implemented in 2007 (Based on Hunt et al. 2008). Petitioner (Not Federal Employee) Requirements for importation of field collected nonindigenous organisms Submit PPQ 526 Permit Application & Permit to Move Live Plant Pest or N oxious Weed to USDA APHISPPQ Plant Health Programs, RIPPS, PPB Issuance of Permit PPQ 526 for containment of Biological Control Agent (BCA) in federal quarantine facilit y Port of Arrival: Removal of contamination Confirmation of identity Confirmation of purity Petitioner: Requirements for release of BCA from containment facility to environment Submit petition following requirement described in RSPM#12 Guidelines for Petition for First Release of Exotic Entomophagous Biological Agents (NAPPO 2008) USDA APHIS PPQ: Review and evaluate petition No NAPPO Consultation: Submit comments and recommendations to USDA APHISPPQ Does USDA APHISPPQ recommend release? Application denied Yes USDA APHIS PPQ : Issuance of Letter of Approval BCA may be moved in accordance with State Laws and Regulation Consult with U.S. Fish Wildlife Service
84 CHAPTER 4 RISK COMMUNICATION DURING THE IMPORTATION AND RELEASE OF ENTOMOPHAGOU S B IOLOGICAL C ONTROL A GENTS IN THE U.S. IS THERE ROOM FOR IMPROVEMENT? Classical biological control can be a key component in invasive species management programs. During the last century, many programs successfully used biological control agents (BCAs) against arthropod pests (van Lenteren et al. 2006) However, when successful, the establishment of these agents is irreversible and, once in a new area, they are very difficult if not impossible to eradicate ( Simberloff & Stiling 1996; van Lenteren et al. 2006) Therefore, decisions to approve the importation and release of any new BCA must be considered carefully by appropriate authorities, particularly taking into account the likelihood of occurrences and consequences of any nontarget effects. This should be done by conducting a pest risk analysis (PRA) (IPPC 2004; IPPC 2005). A PRA summarizes the available scientific evidence in order for a decision to be made on the importation and release of entomophagous BCAs ( Bar r att & Moeed 2005). According to Fisher et al. (1994) a PRA is comprised of risk assessment, which estimates the likelihood of occurrence of a hazard and the magnitude of the consequences; risk management, which identifies options to mitigate the consequences of adverse events; and risk communication, which involves two way exchange of information between decisionmakers and stakeholders. At the international level, the International Standard for Phytosanitary Measures ( ISPM ) #2 Framework for Pest Risk Analysis describ es PRA as having three stages: risk initiation (which involves the identification of pests or pathways of concern), risk assessment, and risk management (IPPC 2007).
85 Regulations for import and release of entomophagous BCAs in the U.S. require applicants to obtain a Federal permit (PPQ 526) for: (1) Importation of BCAs into containment facilities; (2) domestic movement of imported BCAs to other containment facilities; and (3) BCA release into the environment (Figure 41). Furthermore, if the BCA has not been previously released in the U.S., the application must be accompanied by a dossier which describes the justification for the proposed action. The dossier must also provide information on the biology and ecology of the pest and the BCA, the economic an d any potential detrimental environmental impacts of the BCA as well as possible mitigation options ( Hunt et al 200 8; Mason et al. 2005) The format of the dossier can be based on the Regional Standard for Phytosanitary Measures # 12, Guidelines for Pet ition for Release of Entomophagous Agents for the Biological Control of Pests, developed by the North American Plant Protection Organization (NAPPO 2008). The International Standard for Phytosanitary Measures # 3, Guidelines for the Export, Shipment, Im port and Release of Biological Control Agents and Other Beneficial Organisms developed by the International Plant Protection Organization also provides guidance on the importation and release process for biological control practitioners in the U.S. (IPPC 2005). In addition, ISPM # 2, Framework for Pest Risk Analysis and ISPM # 11, Pest Risk Analysis for Quarantine Pests including Analysis of Environmental Risks and Living Organisms give information on the risk analysis process (IPPC 2007; IPPC 2004). In several countries such as Australia and New Zealand, a risk communication framework is defined and integrated within the PRA process during the permitting process of entomophagous BCAs (Hunt et al. 200 8). For instance, in Australia, public
86 comments a re solicited during the importation approval process and during the environmental release of the BCAs ( Hunt et al. 2008). In New Zealand, petitioners have the opportunity to defend their application in a court like setting which might consist of relevant governmental decisionmakers, experts, stakeholders, and members of the general public (Hunt et al. 200 8). In the U.S., prior to 2007, environmental assessments were only required for federal employees and the information needed to be published in Feder al Registry for public comments. However, the importance and role of risk communication is still a poorly defined concept for many federal and state agencies in the U.S. ( Walls et al. 2004). Risk communication can be described as a consensus structure that joins the interests and needs of both senders and recipients ( Fischnoff 1990). Within a PRA framework, senders include researchers and other permit applicants who may work for Universities, Federal or State governments, or the private sector. Sender s are the ones that provide information on the potential risks associated with the importation and release of entomophagous BCAs. Recipients are governmental entities that regulate the permitting process. The National Research Council ( 1996) defines ri sk communication as an interactive process of exchange of information and opinions among individuals, groups, and institutions. Therefore, the main purpose of risk communication is to provide individuals with enough information to enable them to make an informed decision about a potential risk ( Gibson 1985; Fischnoff 1990; Gow & Otway 1990) Governmental agencies have long been concerned with communication methods used to convey the risks associated with environmental issues ( Chess et al. 1995). The US DA APHIS has a long history of evaluating stakeholder satisfaction with its risk
87 communication efforts (Fis her & Chen 1996) It was one of the first agencies within the USDA to conduct a baseline survey of a wide range of stakeholders to examine how well they were meeting their stakeholders needs during risk communication activities ( Fisher & Chen 1996). In 1995, the now defunct USDA APHISPPQ, National Biological Control Institute (NBCI) identified areas in the current biological control regulatory syst em that could be improved. One of those areas was customer service and communication during the permitting process of BCAs ( APHIS 1996 ). Unfortunately, with the demise of NBCI, recommendations made to improve risk communication by USDA APHISPPQ were not implemented. In 2006, a follow up internal evaluation of the USDA APHIS PPQ entomophagous BCAs permitting process revealed that customer service (stakeholder communication) remained an activity that should be improved ( APHIS 2006). In recent years, there has been an increased interest in risk communication among government agencies in the U.S. ( Chess et al. 1995; Fisher & Chen 1996; APHIS 2009 ). However, there has been a lag in implementation of risk communication practices during the permitting process for the importation and release of BCAs ( Messing 2005; Hunt et al. 200 8 ) In this study, areas of risk communication practice are identified that could be improved in order to enhance the current permitting process in the U.S. Also, how USDA APHISPPQ st akeholders receive information on risks pertaining to the permitting process of entomophagous BCAs and how they viewed the agencys risk information activities and performance are described. Based on the results, recommendations for improving risk communi cation practices during the BCA permitting process in the U.S. are suggested.
88 Materials and Methods The data described below were collected by means of a webbased questionnaire. A modification of the mental models approach was used to develop the ques tionnaire. The mental models approach developed by Morgan et al. ( 200 2) identifies gaps and misconceptions on critical problems from both the target audience and the experts, by gathering information from both groups. In our study, t h e approach involved a series of five steps First, an expert model was created (Figure 41) (based on a survey of the literature) to determine the nature and magnitude of the risk communication problem during the permitting process. Subsequently, the expert model was used to c onduct interviews with a small group of knowledgeable stakeholders in order to elicit their perceptions of the risk communication deficiencies and problems. Then, a confirmatory questionnaire was developed (Appendix F) and a quantitative survey was c onducted and administered to an expanded group of stakeholders in order to estimate the prevalence of the identified beliefs. We identified areas in the current risk communication framework (during permitting of BCAs) that needed improvement. The last st ep in the mental models process was not covered in the present study. This step involves the evaluation of the improved risk communication framework to assess its efficiency and practicality with permit petitioners. A committee comprised of 30 experts fro m various agencies and backgrounds, including risk analysts, academic researchers, and members of the private sector, was assembled. Individual members were selected based on their experience and knowledge about the USDA APHISPPQ permitting process and r isk communication procedures. The Dillman method (Dillman 2000 ) was used to develop and administer a survey of 15 openended questions ( Appendix C ). This method attempts to maximize
89 response rates by minimizing the cost of responding, while establishing trust with the respondents. Openended questions were designed to generate perspectives from committee members on the risk communication practices of USDA APHISPPQ and the critical points that should be targeted during the risk communication process. In accordance with the Dillman method, a personalized letter of notice was sent to the selected committee members explaining the goals of the study, the reason for their inclusion in the expert committee, and the reason for sending them the questionnaire (Ap pendix A). Approximately one week later, each participant received the questionnaire with a cover letter (Appendices B and C). A follow up notice was sent a week later thanking those participants that had already responded and requesting a response from those who had not yet responded. Two weeks later a reminder was sent to nonrespondents (Appendix E). Based on the results of the survey, topics and priorities were identified and addressed in a confirmatory questionnaire. A second w ebbased questionnai re comprised of 18 close ended questions was developed (Table 41; Appendix F). Each question was reviewed and pretested (by graduate committee members) to ascertain its clarity. This study used 18 questions of which 16 were subjected to statistical analysis, and 2 questions, requesting general information (Q1, 2), were not included. The first question requested background information (name, company affiliation, and contact information) about the respondent. In addition, respondents were asked about the following: their involvement in biological control (Q 3), their frequency of risk communication in context of their profession (Q 4), the importance of risk communication (Q 5), the sources and frequency of risk communication (Q 6, 7), their satisfaction with risk communication from and interactions
90 with USDA APHISPPQ (Q 8, 9), channels for risk communication (Q 10), their ranking goals for risk communication (Q 11), effectiveness of USDA APHISPPQ in fulfilling communication goals (Q 12), their familiar ity with guidance documents (Q 15), the adequacy of USDA APHIS PPQ website (Q 16), and their knowledge on who to contact (Q 17). The last question (Q 18) solicited additional comments from the respondent. A combination of several databases and directories (e.g. Government agency staff, university faculty, and professional societies) were used to compile a list of 500 decisionmakers and stakeholders. Different words used during the search for potential respondents included biological control, entomolog y, regulatory entomology and quarantine. A modification of the Dillman method (Dillman 2000) was used to administer the webbased questionnaire. An introductory message was sent along with the webbased questionnaire. In addition, a note thanking r espondents for their participation was automatically sent with the webquestionnaire. An electronic message reminding those who had not responded was sent 2 weeks later with the electronic link to the websurvey. The respondents were grouped into five categories based on their affiliation as follows: federal, state, university, nongovernmental agency, and private sector. The Kruskal Wallis test (H value) was used to assess pairwise comparisons of groups of stakeholders that had an independent distribution with ordinal and rating responses as used in the survey (Sokal & Rohlf 1981). This nonparametric test also was used to determine whether the distributions of the responses were statistically different across the different groups of stakeholders.
91 Results Response Rate and R espondent C haracterization Out of the 500 webbased questionnaires sent, 105 participants responded, 29 were undeliverable due to incorrect email addresses, and 5 optedout from participating. An adjusted response rate of 23% was determined. Responses to the websurvey mostly came from participants involved in research (62%) 92% from the university group, 49% from the federal group, and 57% from the state category (Table 41). A smaller percentage of the respondents (19% ) were involved in regulatory aspects during the implementation of biological control programs (30% from the federal group and 36% from the state group). Less than 4% of the respondents were involved in commercial production of BCAs (Figure 42). Import an ce of Risk Communication The majority of respondents across the three major types of affiliations (university researchers, federal, and state employees) considered risk communication to be an important component during the permitting process of entomophagous BCAs (Table 41). Participants from the private sector were evenly divided on the importance of risk communication (Table 41). Risk C ommunication F ramework Four diagrams showing the relationship between risk analysis, risk management, and risk communication were presented in the questionnaire (Figure 43). In various literature sources (CFIA 2000; APHIS 2007 ; IPPC 2007), risk communication is integrated within risk analysis, and illustrated as an independent processes interconnected to the risk assessment and risk management elements (Model A, Figure 4 3). When respondents were asked which of the four diagrams best described existing
92 risk communication pract ices, 34% of the respondents considered Model A to be the best representation. The federal and state group chose Model A (36%, 31% respectively) or Model D (27%, 38% respectively) whereas the university group selected Model A (33%) or Model B (37%). Within the private sector category, Models B and D received the same level of significance (50%). Frequency and S ources of Risk Communication Nearly 80% of respondents indicated that they communicated about risks in the context of their professions monthly or more frequently (Table 41). Specifically, more than 80% of researchers across the different types of affiliations communicated at least monthly about risk in the context of their profession (Figure 44). Nearly 70% of federal and state regulators indic ated they had weekly communications about risk (Figure 44). To accomplish these risk communication activities, respondents relied on a combination of traditional communication channels such as faceto face meetings, telephone exchanges, televised programs pamphlets, and scientific publications, and electronic communication channels, such as e mails, list servers, Federal Registry site, and blogs (Table 41). Most of the risk communication information received by stakeholders was conveyed by USDA APHISPPQ ( 29.9 %) university researchers (2 8 .9 %), and state and local plant protection agencies ( 21.6 %) ; with less information from environmental groups (15.5%) and the Cooperative Extension Service (10.3%) (Figure 4 5). The majority of stakeholders received inf ormation from USDA APHISPPQ once a year or less frequently (72.6%). Environmental groups followed by Cooperative Extension Service personnel were the least involved in the transfer of risk information (17.4% and 15.4% respectively) (Figure 45)
93 Goals of R isk C ommunication Overall, the respondents believed that explaining the risks associated with the importation and release of entomophagous BCAs should be the most important goal of risk communication activities (mean score 3.64, Figure 46). On the other hand, they also believed that one of the main objectives of these interactions should be to explain the decisions made during the importation of entomophagous BCAs (mean score 3.29). In decreasing importance, they considered that the process should encourage good practices among biological control practitioners (mean score 3.06), respond to external peer review recommendations (mean score 2.88), and explain the different petition requirements needed during the importation of process for BCAs (mean score 2.13). Based on the analysis with Kruskal Wallis, there was a statistical difference in the way the various groups of stakeholders ranked the key goals of risk communication (H = 12.5; 4 d.f.; P = 0.01). Respondent S atisfaction with R isk I nformation and I nteractions The respondents were somewhat familiar with which entities to contact during the permitting process (Table 41). About one third of respondent s across the different groups were satisfied with quality of the content of the risk message (30.0% Figure 47 ) and with the risk communication exchanges and interactions (26% Figure 48 ) they received from USDA APHIS PPQ. When risk communication interactions occurred between USDA APHISPPQ and their stakeholders, analysis with Kruskal Wallis demons trated that federal, state and the university groups ranked the Agencys effectiveness in fulfilling risk communication goals the same way (H = 5.1; 4 d.f.; P = 0.3) (Figure 49). In contrast, professionals from the private sector (60%) believed the
94 Agenc y to be ineffective in explaining the risks pertaining to the importation of entomophagous BCAs (Figure 49). Need for more G uidance D ocuments The federal and state respondents were somewhat familiar with international and regional standards for phytosanitary measures related to pest risk analysis or specific to importation and release of BCAs. Private sector respondents were unfamiliar with the various ISPM and RSPM guidance documents (Table 41). Although the USDA APHISPPQ website provides some infor mation, the respondents recognized the need for more information from USDA APHISPPQ focusing on the risks pertaining to the importation and release of entomophagous BCAs (Table 41). Public Involvement Respondents from state and university groups felt that biological control stakeholders were not appropriately included during the decisionmaking process of the permitting of arthropod BCAs (Table 41). Discussion To date, there has been relatively little attention given to understanding risk communication activities during the permitting process for the importation and release of entomophagous BCAs in the U.S. Therefore, although risk communication is an important component of the PRA process, it is still an ambiguous concept for many Agency professional s and their stakeholders ( Walls et al. 2004). There is a need, therefore, to identify ways in which risk communication can be improved and thus lead to the development of an improved framework that will satisfy the needs of stakeholders. This new framework would address some of the key concerns expressed
95 by biological control practitioners, environmental groups, and the general public (Thomas & Willis 1998; Simberloff 2005). One area that should be clarified is how risk communication is currently integrat ed within the PRA framework. Although Models A or B (Figure 42) were more frequently chosen by respondents (68.2%), various scientific publications and many respondents (Table 41) indicated that Model D (Figure 43) best described how risk communication should be integrated. In Model D, risk communication is an integral element of risk assessment and management components within the PRA. The difference in opinions between what is the current practice and what should be targeted demonstrated a flaw in th e current risk communication framework. Although Model D may seem unobtainable for a novel risk communication framework, it should form a basis for a more participatory based PRA model. Consultations between the general public or experts and USDA APHIS P PQ during the permitting process for BCAs should provide an additional source of knowledge to validate the identification of risk factors and management options. Although one of the current concerns of USDA APHISPPQ is to increase public involvement in d ecision making process (APHIS 2009) there is lack of information on r isk communication activities by the Agency Previous studies showed that stakeholders had little knowledge of the risk analysis framework pertaining to the importation and release of entomophagous BCAs, consequently limiting their participation (APHIS 200 6; APHIS 2009). This study showed that stak eholder perception and understanding of the process, the communication channels used, and the efficiency of the risk message should be improved in order to increase participation
96 by stakeholders In addition, the survey showed that stakeholders received i nformation from only a few sources and that the information was received very infrequently. Only 1% of respondents thought that the USDA APHIS PPQ website was efficient at providing guidance. It seems that USDA APHIS PPQ is aware of these issues. Indeed, in November 2009, the Agency conducted a survey of their registered stakeholders to obtain feedback on how they could improve the overall delivery of information on their website ( APHIS 2009). The development of website links relating to critical issues relating to PRA will provide improved information and guidance to stakeholders. Our study showed that n ew channels of communication should be investigated to increase stakeholder access to risk related information. This might include the use of televisi on, national public radio, or newspaper during the communication of risk. Even when stakeholders received information from USDA APHISPPQ, this message did not always meet their needs For instance, u niversity and p rivate sector s respondents said that US DA APHISPPQ was ineffective in communicating risk pertaining to the importation and release of entomophagous BCAs (Fig ure 49). The difficulty faced by USDA APHISPPQ in fulfilling stakeholder needs may come from the fact that different groups of stakeholders view risk communication goals differently (Figure 4 6). Therefore, there is a need to identify the main goals of the risk communication efforts, specific to the different types of stakeholders and respond accordingly. A majority of the respondents w as not satisfied with the quality of risk the communication messages or the risk message exchanges and interactions from USDA APHISPPQ (Figure 4 7). For instance, 60% of respondents from the private sector
97 were either dissatisfied or very dissatisfied wi th the risk communication messages and interaction with USDA APHISPPQ (Figure 4 8). This result seemed to confirm the negative perception of USDA APHISPPQs customer service record from their stakeholders as illustrated by Warner & Getz (2008). A great er level of involvement in the decisionmaking process by stakeholders and expert peer review groups may increase the stakeholders trust in the decisions and improve the stakeholders perception of the quality of the risk communication message. In October 2009, a proposed rule was submitted by USDA APHISPPQ for the mandatory development of an Environmental Assessment (EA) before the importation of entomophagous BCAs (APHIS 2009). Under the National Environmental Policy Act (NEPA), the development and submission of an EA is required when a proposed action such as the introduction of any organism has potentially significant environmental impacts ( Kubasek & Silverman 2005). An external group of experts selected by the governmental agency reviews the EA. T he group of experts then provides an analysis of potential adverse environmental effects of the proposed action. In accordance with the Administrative Procedure Acts rules on informal rule making, a draft is published in the Federal Register and public comments are accepted by the stakeholders for a period of 60 days. One of the major advantages of this process is that it requires public participation in the decisionmaking process. In addition, the development of a standardized risk communication framework with clear and identified risk communication activities will increase the quality of the interactions between the agency and its stakeholders. The results from this survey provide baseline data to evaluate USDA APHISPPQs risk communication performance during the importation and release of
98 entomophagous BCAs. Based on the findings of this study, the following are being suggested to enhance the risk communication framework: increase of transfer of guidance documents and information pertaining to PRA process of entomophagous BCAs through the use of additional media; greater involvement of cooperative extension faculty in stakeholders education about PRA; identification and development of risk communication messages specific to different types of stak eholders; development of a PRA framework with a detailed time frame which will increase stakeholder involvement in the decisionmaking process.
99 Table 41. Questionnaire. Questions 1. About Yourself 2. In which group will you categorize yourself? 3. How would you categorize your involvement in biological control? 4. How often do you communicate risk in the context of your profession? 5. Do you view risk communication as an important component during the importation process of entomophagous BCAs? 6. From which entity(ies) do you receive information pertaining to risks associated with importation of BCAs and what is relative importance of each source? 7. How often do you receive information about risks associated with the impor tation of entomophagous BCAs from USDA APHISPPQ 8. How would you rate your level of satisfaction with the risk communication information that you perceive from USDA APHIS PPQ pertaining to the importation of entomophagous BCAs? 9. How would you rate you r level of satisfaction of the Risk Communication interactions with USDA APHISPPQ concerning the importation of entomophagous BCAs? 10. What percentage best describes the communication channel(s) through which you receive the information on risks pertaining to the importation of entomophagous BCAs?
100 Table 41. Continued Questions 11. Rank the following key goals of the risk communication process during the importation of entomophagous BCAs in order of importance 12. How effective is USDA APHIS PPQ in fulfilling each risk communication goal during the importation of entomophagous BCAs? 13. What is your degree familiarity with the different guidance documents pertaining to the importation of entomophagous BCAs? 14. Which of these mode ls best represent your perception of risk communication as it is currently incorporated during the importation process of entomophagous BCAs 15. Do you think there is a need for more guidance documents from USDA APHISPPQ concerning the importation of entomophagous BCAs? 16. Does your USDA APHIS PPQ website provide you with enough explanations and guidance about importation of entomophagous BCAs? 17. Do you have the information (phone numbers, emails, fax number, address) of points of contact that you can reach if you have any questions during the importation of entomophagous BCAs process? 18. In your opinion, is the public adequately involved in the importation of entomophagous BCAs process?
101 Table 42 Summary of q uestions and responses obtained from 5 categories of biological control stakeholders (Federal, State, University, Private, and Non Governmental). Question Stakeholder Categories a Federal (40%) State (18%) University (34%) Private (6%) 3. Biological control area of involvement Research (57%) Regulation (24%) Regulation (53%) Research (42%) Research (92%) Commercial production (67% Conservation (17%) 5. Is RC b important? Yes (92.5%) Yes (90%) Yes (100%) Yes (33%) No (33%) 7. RC Frequency from PPQ c At least yearly At least yearly Yearly to never (91%) Never (60%) 10. Main RC channels Mailed letters Scientific publications Scientific conferences emails Scientific publications Scientific conferences Scientific publications Scientific conferences emails Meetings (lunch, social, or board) 13. Familiarity with guidance documents Somewhat to very familiar Somewhat familiar to familiar Unfamiliar to somewhat familiar Unfamiliar 15. Need for more guidance documents Yes, mostly to definitively (60%) Yes, somewhat to mostly (65%) Yes, mostly to definitively (56%) Yes, definitively (100%) 16. Is PPQ website provide enough guidance Yes, somewhat to mostly (54%) Not at all to yes, somewhat (65%) Yes, somewhat (68%) Yes, somewhat (100%) 17. Knowledge of point of contacts Yes, somewhat to mostly (57%) Yes, somewhat to mostly (59%) Yes, somewhat to mostly (85%) Yes, somewhat (100%) 18. Is public involvement adequate? Yes, somewhat to mostly (53%) Not at all to undecided (52%) Yes, somewhat to not at all (74%) Yes, somewhat to undecided (100%) aNonGovernmental Organization less that 2% bRisk Communication cPlant Protection and Quarantine Organization
102 Figure 41. Expert model of the p ermitting process used by USDA APHISPPQ in 2007 (Hunt et al. 2008). Petitioner (Not Federal Employee) Requirements for importation of field collected nonindigenous organisms Submit PPQ 526 Permit Application & Permit to Move Live Plant Pest or Noxious Weed to USDA APHISPPQ, Plant Health Programs, RIPPS, PPB Issuance of Permit PPQ 526 for containment of Biological Control Agent (BCA) in federal quarantine facility PORT OF ARRIVAL: Removal of contamination Confirmation of identity Confirmation of purity Petitioner: Requirements for release of BCA from containment facility to environment Consult with U.S. Fish Wildlife Service Submit petition following the guidelines provided by RSPM#12 Guidelines for P etition for First Release of Exotic Entomophagous Biological Control Agents (NAPPO 2008) USDA APHIS PPQ PHPRIPPS PPB : Review and evaluate petition No NAPPO Consultation: Submit comments and recommendations to USDA APHISPPQ Does USDA APHISPPQ recommend release? Application denied Yes USDA APHIS PPQ : Issuance of Letter of Approval BCA may be moved in accordance with State Laws and Regulation
103 Figure 42. Distribution of respondents to Question 1: In which group will you categorize yourself? Figure 43. Different model choices of p est r isk a nalysis structure presented in questionnaire. RA = risk analysis, RM = risk management, RC = risk communication.
104 Figure 44. Distribution of respondents to Question 4: How often do you communicate risk in the context of your profession? Figure 45. Distribution of respondents to Question 6: From which entity(ies) do you receive information pertaining to risks associated with importation of BCAs and what is the relative importance of each source (percentage)?
105 Figure 46. Distributio n of respondents to Question 11: Rank the following key goals of the risk communication process during the importation of entomophagous BCAs in order of importance (5very important to 1least important). Figure 47. Distribution of respondents to Qu estion 8: How would you rate your level of satisfaction with the risk communication message/ information that you receive from USDA APHIS PPQ pertaining to the importation of entomophagous BCAs?
106 Figure 48. Distribution of respondents to Question 9: How would you rate your level of satisfaction with the risk communication exchanges/ interactions that you receive from USDA APHIS PPQ pertaining to the importation of entomophagous BCAs? Figure 49. Distribution of respondents to Question 12: How effective is USDA APHISPPQ in fulfilling each risk communication goal during the importation of entomophagous BCAs?
107 CHAPTER 5 COLLABORATIVE RISK ASSESSMENT DURING THE PERMITTING PROCESS OF ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS A BETTER PROCESS? The task of protecting American agriculture and natural resources against the risks associated with the entry, establishment, and spread of plant pests in the U S is the responsibility of the U.S Department of Agriculture (USDA) Animal and Plant Health Ins pection Service (APHIS) Plant Protection and Quarantine (PPQ) (APHIS 2011) the U.S. National Plant Protection Organization (NPPO). The Plant Protection Act (PPA 2000) authorizes the Secretary of Agriculture to delegate authority for plant protection authority to USDA APHIS PPQ This federal entity has broad jurisdiction to develop and enforce phytosanitary measures that will prevent and or delay the introduction and spread of plant pests ( PPA 2000 ) In the 1990s, scientists and gov ernments across the globe identified a need for harmonization of regulatory and phytosanitary procedures during the permitting process for entomophagous biological control agents ( BCAs ) in order to ensure more effective plant and animal protection ( Ebbels 2003) However, regulations and legislation vary considerably between countries (Chapter # 3). In an effort to harmonized PRA processes and also provide procedures to assess risks associated with the importation and release of BCAs, the International Plant Protection Convention (IPPC) developed the International Standard for Phytosanitary Measures (ISPM) # 2 Framework for Pest Risk Analysis (IPPC 2007). N ew and revised legislation and regulations were implemented by the U.S. NPPO in response to a need for a comprehensive, harmonized regulatory process for the importation and release of BCAs (De Nardo & Hopper 2004). A major addition of the Plant Protection Act of 2000 was USDA APHISPPQs statutory authority to regulate organisms intended to control pant pests. Prior to this Act, a legal definition of a biological control agent was not
108 present in the statutes of the U.S., and the U.S. NPPO regulated them under statutes and regulations pertaining to plant pests. In 2001, following an agreement with Can ada and Mexico, the U.S. NPPO, under the North American Plant Protection Organization (NAPPO), requires a permit for environmental release of an entomophagous BCA (NAPPO 2008). One of the priorities of USDA APHISPPQ has been to improve their understanding of the potential risks associated with the import ation and release of entomophagous BCAs Decisions for approving or denying the importation of BCAs are made by conducting a pest risk analysis (PRA) ( Hunt et al. 2008; Mason et al. 2005). Risk based analysis provides a structured framework for making rational decisions when outcomes are uncertain (Simberloff 2005). One of the main objectives of a PRA is to confirm absence of contamination and identification of BCAs or associated organisms whose introduc tion could lead to detrimental agricultural/environmental impacts as well as to identify those BCAs that pose little risk (Page 1978). Quality control of BCAs during consignment is an important step because it establishes presence of harmful contaminants associated with the imported BCAs. The permit petitioner is responsible for providing information on the BCA following the format recommended by the Regional Standard for Phytosanitary Measure # 12 Guidelines for Petition for First Release of Exotic Entomophagous Biological Control Agents (NAPPO 2008). Information about the proposed action, biology and ecology about the target pest and BCA, potential economic and environmental impacts are required. The science based PRA reduc es the scientific uncertainty associated with potential risks by examining information helpful in evaluat ing management options (DeK ay et al 2002)
109 Uncertainty is inherent to all risk analyses (Koch et al. 2009). Uncertainty can be categorized into epistemic, linguistic, and stochastic (natural variability) (Burgman 2005). Epistemic uncertainty is generally described as a shortfall in knowledge which may lead to undesired outcomes (DeKay et al 2002) Pest risk analysis is mostly affected by epistemic uncertainties (Rafoss 2003; K och et al. 2009). The p recautionary principle is sometimes used during decision making when there is uncertainty (Graham 2001) and was accepted as a valid approach by the Rio Convention on Biodiversity in 1992 (Simberloff 2005). The principle states that precautionary measures are authorized for environmental decisions even when a causal relationship has not been fully establis hed (Simberloff 2005). The principle is often invoked in complex environmental situations where detrimental impacts are often irreversible and losses difficult to recover. Therefore, the precautionary approach is more likely to be used in cases where tra ditional regulator y approaches are inadequate (Dek ay et al 2002) and there is no consensus about the likelihood of occurrence of potential detrimental impacts (Simberloff 2005). The precautionary approach ensures that the absence of scientific certainty is not used as a reason for postponing actions that are intended to protect people and the environment (Arrow & Fisher 1974; Dixit & Pindyck 1994; Farrow & Morel 2001). For intentional introduced non native species such as entomophagous BCAs, rigorous quarantine laws adopting a guilty until proven innocent approach are believed to be used in the U.S. (Ruesink et al. 1995, Simberloff 1996, 2005). This approach is justified by the fact that in the event of unintended environmental impacts from an authori zed release of an entomophagous BCA, the U.S. NPPO will be the responsible party not the researcher. Several scientists argue that such approach is
110 used as justification for arbitrary denial of entomophagous BCAs permits. However, there has been recognit ion that a level of zero risk in the case of BCA is unobtainable and that a certain level of risk is acceptable considering the benefits. The lack of transparency in the process of assessing data during the permitting process for entomophagous and phytop hagous BCAs by USDA APHISPPQ creates additional distrust among stakeholders in the current risk assessment process (Mason et al. 2005; Warner & Getz 2008). In this study, the hypothesis tested is that credibility in the current risk analysis process can be improved through the development of a collaborative approach between decisionmakers and stakeholders. Collaboration involves the process of joint decisionmaking among key stakeholders, experts and decisionmakers (Daniels & Walker 2001). Decisions r esulting from a collaborativebased r isk analysis m odel are more widely accepted by participants because they provide legitimacy to environmental decisions (Fischnoff 2005a,b) It builds trust among participants and allows a balanced review of information during the r isk assessment process (Fischnoff 2005a,b) The mental models approach (Morgan et al. 2002) was used to identif y gaps, misconceptions and critical problems in participant s comprehension of the risk analysis process by gathering information f rom both decision makers and stakeholders. The approach is based on a systematic analysis of participants beliefs and the identification of knowledge gaps during the decisionmaking process (Morgan et al 2002) Based on the findings, a modified collaborative based risk analysis process was developed.
111 Materials and Methods Committee of Experts A committee of 30 experts with various backgrounds covering five professional affiliations (federal, state, university, environmental and the private sector) was chosen (Table 51). A combination of several databases and directories (e.g. government agency staff, university faculty and professional society) was used to compile the committee of experts. The members of the expert committee were selected based on t heir experience and knowledge concerning the permitting process for BCAs and risk assessment procedures used to evaluate entomophagous and phytophagous BCAs for importation and release. All of the experts had submitted at least one petition for importation and/or release of a potential entomophagous BCA or been actively involved in a classical biological control project that involved importation and release of natural enemies. Development of an E xpert Conceptual M odel and Interview Protocol The mental models approach (Morgan et al. 2002) was used to compare and analyze the different beliefs held by the group of experts. The mental models approach consists of a series of 5 steps described as follows: 1development of an expert conceptual model based on scientific data to assess issues associated with the risk analysis process; 2administration of interviews to elicit stakeholders beliefs; 3creation of a confirmatory questionnaire to evaluate the proportion of stakeholders which share the same beliefs; 4 correction of misconceptions; and 5development of an improved process based on findings. A conceptual diagram was developed based on information from scientific publications and the USDA APHISPPQs website (Figure 41, Chapter 4). The
112 diagram capture d the different steps involved in the permitting process for entomophagous BCAs when submission was made by a nonfederal employee. The conceptual diagram was used to assess the groups understanding of relationships and interaction of key factors during the risk analysis process of entomophagous BCAs and to identify speci fic issues affecting the efficiency of the permitting system Openended questions directed participants to specific topics in the expert conceptual model. A modification of the Dillman method (Dillman 2000 ) was used to develop a semi structured interview with 15 openended questions (Appendix C). The questionnaire can be found in Table 41. The open ended questions were designed to characterize settings or events that might affect the operation and efficiency of the risk analysis framework. The questionnaire was reviewed and pretested to ascertain the clarity of each question. Comparison of Mental Models for Decision Makers and Stakeholders Results from the interviews allowed for t he characterization of themes and patterns of issues related to the risk assessment process. They were used to compare the beliefs of decisionmakers and stakeholders. Findings were summarized and used to identify the c ommonalities and divergences among participant s. Recommendations were used to develop a suggested collaborative risk analysis process to be used during the importation and release of entomophagous BCAs. Results and Discussion Conceptual Expert Model Not surprisingly, the interview result s generated different mental models for decisionmakers and stakeholders on issues impacting the permitting process for entomophagous BCAs (Appendix D). The mental models for each group were
113 compared and categorized into eight categories, which corresponded to crucial components of the risk analysis process (Table 52). These components were described as follows: risk assessment, risk communication, submission of a petition for importation of entomophagous BCA, inspection at the port of entry, submission process for petition to release a BCA, external review process, decisionmaking process, and issuance of letter of approval for release. Problem areas pertaining to the risk analysis process during permitting were characterized into five categories (risk assessment process, stakeholder participation, risk communication, decisionmaking, and selection of expert group members during decisionmaking process) (Table 52). Comparison of Mental Models for Decision Makers and Stakeholders by Categories Risk anal ysis process Stakeholders (permit petitioners) believed that a guilty until proven innocent approach was used during the risk analysis process and that it was often used by USDA APHISPPQ decision makers as a justification to prevent the importation or r elease of candidate entomophagous BCAs. Decisionmakers argued that due to high uncertainty related to complex environmental scenarios for entomophagous BCAs, an innocent until proven guilty risk analysis approach could not be used. They pointed out that there was often a lack of transparency in the information provided by the permit petitioner. In addition, they indicated that there was no accepted standardized process to quantify uncertainty during their risk analysis process. The precautionary principle postpones taking actions that may result in irreversible outcomes when there is a prospect for obtaining improved information (Simberloff 2005). Uncertainty is often described as a gap of knowledge or as lack of, incomplete, erroneous, dated, or
114 ina ccurate knowledge. However, Wynne (1992) defined s cientific uncertaint y as a subjective combination of complex social and cultural factors Therefore, uncertainties can never be fully resolved before taking action (Graham 2001) In addition, the precaut ionary approach is always appropriate for environmental decisions because r isks are often not demonstrated until later when damages have already occurred and are irreversible (Graham 2001). The modified risk analysis process (Figure 51) proposed in this study is based on a collaborative model. A collaborativebased process is a t wo way interaction model that considers concerns and information from both key stakeholders and decisionmakers. It also assumes shared responsibility for subsequent outcomes from those actions (Selin & Chavez 1995) unlike in the current risk approach used for permitting entomophagous BCAs in the U.S. Stakeholder participation Stakeholders often felt that the risk analysis process during entomophagous BCA permitting was subjective and arbitrary. They believed that the general public should be more involved in the decisionmaking process in order to increase transparency in the process. Participation is defined as a process where individuals, groups and organizations choose to take an active role in making decisions that affect them (Wanderman 1981; Wilcox 2003; Rowe et al. 2004). T he complex and dynamic nature of environmental problems requires flexible and trans parent decisionmaking procedures that embrace a diversity of expertise ( Stringer & Reed 2007 ). To achieve this, stakeholder and/or expert involvement needs to be increasingly embedded into environmental decision making process (Stringer & Reed 2007) Th e modified collaborativebased risk analysis process (Figure 51) requires the development of an Environmental Assessment (EA). The EA document provides basic information on the
115 positive and negative environmental impacts of the proposed action, including a management program in case of adverse consequences, and also suggests alternatives to the implementation plan under consideration (NEPA 1970). A similar approach is recommended by the Regional Standard for Phytosanitary Measures # 12 (Guidelines for P etition for First Release of Exotic Entomophagous Biological Control Agents) developed by the North American Plant Protection Organization (NAPPO) and the International Standard for Phytosanitary Measures # 3 (Guidelines for the export, shipment, import and release of biological control agents and other beneficial organisms) published by the International Plant Protection Convention (NAPPO 2002; IPPC 2005). However, in accordance with the rules on informal rule making of the U.S. Government Administrati ve Procedure Act, during the development of an EA, the proposed action must solicit public comments for a period of 60 days (Kubasek & Silverman 2005). Risk communication Results from these interviews revealed that a well orchestrated risk communication process is needed in order to reduce information gaps between decisionmakers and stakeholders. Participants considered that risk communication was an important component of the risk analysis process. Results showed that both decisionmakers and stakeholders believed that risk communication should be integrated within this process (Table 52). The role of risk communication in environmental decision making has significantly increased within USDA APHIS over the last decade and remains an important social i ssue (Santos & Chess 2003) It provides a means to determine what matters most to USDA APHISPPQ decision makers (Wardman 2008) However, the majority of the stakeholders interviewed were
116 dissatisfied with the risk communication interactions and messages received from decisionmakers (Table 52). The critical issues identified were: unacceptable communication channels used during risk communication activities, problems with the main source of information, and poor quality of risk communication interactions between decisionmakers and stakeholders. The areas of risk communication that needed improvement are further discussed in detail in Chapter 4 of this dissertation. External review process and selection of expert group members This study showed that stakeholders felt that there was a l ack of external peer consultations during the decision process of issuing permits for entomophagous BCAs. Generally, decisions made after the development of a risk assessment are based on expert judgment (Daniels & Walk er 2001). The stakeholders believed that an independent assessment of the facts by an expert group would provide a more transparent decision than USDA APHISPPQs seemingly arbitrary decisions. An expert can be described as someone with an appropriate degree of knowledge about an issue and that has efficient communication skills (Meyer & Booker 1990). An expert group must be carefully selected because its composition affects the outcome of the risk assessment (Daniels & Walker 2001). For instance, the F ood and Agriculture Organization used a stratification approach for the selection of experts for issues of food safety based on their affiliation, technical expertise, professional recognition, publications, and ability to draft clear reports (Meyer & Book er 1990). The selection of experts for external review could be done in a way similar to the Technical Advisory Group (TAG), an expert group that advises USDA APHISPPQ on risk assessment of importation and release of phytophagous BCAs (Scoles et al. 2008). The TAG is composed of representatives of federal agencies that might be affected by decisions,
117 such as the USDA Agricultural Research Service, USDA Forest Service, USDA Natural Resources Conservation Service, U.S. Environmental Protection Agency, etc (Scoles et al. 2008). In addition, state employees may also be incorporated in the TAG. In the proposed modified risk analysis process for entomophagous BCAs, a representative from the State Plant Regulatory Office (SPRO) could make the connection between the state and federal decisionmakers. Decision Making Process Approval for import to federal quarantine facility In the modified risk analysis process (Figure 51); the permit petitioner must initially contact the USDA APHISPPQ service staff office to discuss the scientific soundness of the proposed action. A consultation with the U.S. Fish and Wildlife Service (FWS) also is suggested at this time rather than later in the process. This first step ensures that possible environmental and economic issues are identified early before the release is made. Based on the initial contact and submitted documentation, risk analysts from USDA APHISPPQ will provide recommendations whether or not to approve the initial importation of the BCA into a quarantine faci lity. No public hearing is required at this initial stage. Approval for environmental release In the modified risk analysis process (Figure 51), all applicants (federal and nonfederal employees) are required to automatically prepare an EA for the envi ronmental release of an entomophagous BCA. Under NEPA, submission of the EA calls for public comments during the decisionmaking process. The EA and the host species list for the assessment of the BCA host range would be reviewed by a newly proposed group of outside experts and recommendations will be sent to risk analysts within USDA APHIS-
118 PPQ. After evaluation of the EA, USDA APHISPPQ would reach a decision based on the recommendations from the expert group and comments from the general public. In addition, recommendations from representatives from NAPPO (Canada and Mexico) also woulb be taken into account during the decisionmaking process. Final decision If USDA APHISPPQ denies the stakeholders permit to release the BCA, a written response is pre pared by USDA APHISPPQ outlining their concerns, justifying their denial, and identifying potential steps to rectify the problems with the EA. The applicant has the option of resubmitting the application after conducting further research which addresses USDA APHISPPQ concerns. After the stakeholder submits supplemental information, USDA APHIS PPQ re evaluates the application, and, in case of continued refusal to permit release of the entomophagous BCA, communicates their reasons to the applicant. If approval for release is granted, USDA APHIS PPQ staff notifies the stakeholder and sends the information to the SPRO in the state where the entomophagous BCA will be released. After communicating with the SPRO, the stakeholder can release the entomophagous BCA following state laws. Summary This study explored how decisions were made and who was involved in making them during the process of approving the importation and release of entomophagous BCAs. Based on the findings, the implementation of a collabor ative based risk analysis process during the permitting process of entomophagous BCA s was investigated. The modified risk analysis process incorporated expert and public participation during the decisionmaking process. In addition, the permit petitioner had the opportunity to re-
119 submit the application if approval for release was denied. Following approval from the federal authority, a direct liaison with a state official was integrated into the process.
120 Table 51. Type of professional affiliation and number of expert committee members. Professional affiliation n University Professor (biological control) 4 Professor (integrated pest management) 1 Professor (plant pathology) 1 Lecturer (regulatory sciences) 1 Researcher (biological control) 2 Researcher (environmental sciences) 1 State Researcher (biological control) 2 Researcher (environmental sciences) 1 Extension specialist 2 Federal U.S. Department of Agriculture Agricultural Research Service (entomologists) 3 U.S. Department of Agriculture Animal and Plant Health Inspection ServicePlant Protection and Quarantine (risk analysts) 6 Private sector 3 Environmental groups 2
121 Table 52 C haracterizing the mental models of decisionmakers and s takeholders during the p ermitting p rocess of entomophagous biological control agents ( BCAs ) in the U.S. Mental m odels Components of the p ermitting p rocess Stakeholders ( p ermit p etitioners) Decision m akers Issue t hemes Recommendations Risk Analysis (RA) p rocess risk analysis process due to misuse of precautionary approach during decisionmaking process for each step of RA process document on RA process o uncertainty in potential irreversible environmental impacts BCAs (Biological Control Agents) follow a precautionary approach appropriate level of environmental protection uncertainty nadequate needs to be increased standardized RA process during decision making process system in case of permit refusal Risk Communication (RC) p rocess RC is to explain risks associated with importation of BCAs ational P lant P rotection O rganization effectiveness in explaining risks to permit petitioners information and interactions with regulators/decisionmakers contacts RC is to explain risks associated with importation and release of BCAs to biological control practitioners and consistency from permit petitioners goals and identify importance of RC activities media outlets to accomplish RC activities International Standard for Phytosanitary Measures (ISPM) on RC or addendum to ISPMs related to PRA
122 Table 52. Continued Components of the p ermitting p rocess Stakeholders (p er mit p etitioners) Decision m akers Issue t hemes Recommendations Submission of petition (PPQ 526) for movement of BCA into Federal Quarantine Facility documents explaining the permitting process at initial stage identified for additional inquiry BCAs due to liability issues APHIS PPQ website and other published articles provide enough guidance information for permit petitionersa permit) create a quicker process contacts for additional information on organism permits are available on USDA APHIS website page process should be clearly defined creation of online links on topics related to entomophagous BCAs documents on relevant information on legislation/ regulations, permitting unit organization, permitting process with corresponding time frame Inspection at the port of entry difficult to fulfill entry (hand carrying) because of lack of instructions and training from Customs and Border Protection (CBP) inspectors from petitioners from permit petitioners regulators and perm it petitioners training on issues pertaining to entomophagous BCAs contacts at port of entry documentation and notification requirements for hand carrying Submission of petition (PPQ 526) for movement of BCA into Federal Quarantine Facility documents explaining the permitting process at initial stage identified for additional inquiry rriers to ship BCAs due to liability issues APHIS PPQ website and other published articles provide enough guidance information for permit petitionersa permit) create a quicker process contacts for additional information on organism permits are available on line process should be clearly defined creation of online links on topics related to entomophagous BCAs documents on relevant information on legislation/ regulations, permitting unit organization, permitting process with corresponding time frame
123 Table 52. Continued Components of the p ermitting p rocess Stakeholders ( p ermit p etitioners) Decision m akers Issue t hemes Recommendations Submission of petition (PPQ 526) for release of BCA into environment petitions/documents petition submission status data requirements tracked online communicate with PPQ staff through emails might followed Regional Standard for Phytosanitary Measures # 12 as recommended in USDA APHISPPQ website ency and reliability of risk assessment process assigned to petitions and in direct contact with petitioner approved petitions for guidance interactions between permit petitioners and decisionmakers External Review review process with North American Plant Protection and Fish and Wildlife Service representatives during decision making process making process is arbitrary and subjective an Expert Group during decision making process Group representing stakeholders interests standardized guidelines for evaluation Decision Making Process appropriate level of environmental safety expertise (scientific and bureaucratic) appropriate level of environmental safety process is inefficient making process is inadequate testing nontarget impacts petition after conducting more research upon recommendations Submission of petition (PPQ 526) for release of BCA into environment petitions/documents petition submission status data requirements tracked online petitioners can communicate with PPQ staff through emails might followed Regional Standard for Phytosanitary Measures # 12 as and reliability of risk assessment process assigned to petitions and in direct contact with petitioner approved petitions for guidance interactions between permit petitioners and decision
124 Table 52. Continued Components of the p ermitting p rocess Stakeholders ( p ermit p etitioners) Decision m akers Issue t hemes Recommendations recommended in USDA APHIS PPQ website makers Issuance of Letter of No Further Regulation/ Accordance with State laws and regulations USDA APHISPPQ to regulate entomophagous BCAs between the State and Federal level permits versus State approvals (2000) gives a broaden authority t o USDAAPHISPPQ to regulate entomophagous BCAs during decision making process is unclear USDAAPHISPPQ and State Plant Regulatory Officials
125 Figure 51. Novel risk assessment process for the permitting process for importation and release of entomophagous BCAs. Steps in gray boxes are additional to existing pest risk analysis process. Petitioner (Not Federal Employee) Requirements for importation of field collected nonindigenous organisms Submit PPQ 526 Permit Application & Permit to Move Live Plant Pest or Noxious Weed to USDA -APHISPPQ USDA APHIS -PPQ Issuance of Permit PPQ 526 for containment of Biological Contr ol Agent (BCA) in federal quarantine facility Port of Entry : Removal of contamination Confirmation of identity and purity Petitioner : Requirements for release of BCA from containment facility to environment Submit an Environmental Assessment or an Environmental Impact Assessment to Expert Group Expert Group Consultation (External Review): Review and evaluate petition and host species list Submit comments and recommendations to USDA APHIS PPQ Does PPQ recommend release? NO NAPPO Consultation (Optional) PPQ Consultation FWS Consultation (External Review) USDA APHIS PPQ Issuance of Permit PPQ 526 for release of BCA in to environment Y ES Public Comments Petitioner : Conducts more research upon recommendations Resubmits Discontinues effort Application denied
126 Figure 51. Continued USDA APHIS PPQ: I ssuance of Letter of Approval Sends permit application with comments and recommendations to the State Plant Regulatory Official (SPRO) BCA may be moved in accordance with State Laws and Regulation SPRO: Review and evaluate petition Issues a permit
127 CHAPTER 6 EGG PARASITOIDS ATTACKING CACTOBLASTIS CACTORUM (LEPIDOPTERA: PYRALIDAE) IN NORTH FLORIDA The cactus moth, Cactoblastis cactorum (Berg) ( Lepidoptera: Pyralidae) is often cited as the perfect example of a successful weed biological control agent ( Moran & Zimmermann 1984). In 1925, the cactus moth was introduced from its native Argentina into Australia to control prickly pear cactus, Opuntia spp which had originally been brought into Australia from Mexico for commercial purposes (Dodd 1940; Mann 1970) The cactus became invasive and made large tracts of rangeland unfit for grazing cattle. Within a few years after the introduction o f C. cactorum into Australia, US $6 million worth of rangeland was restored, equivalent to more than US $60 million in todays dollars (Dodd 1940; Williamson 2009). Based on these promising results, C. cactorum was import ed from Australia to South Africa, Mauritius, and Hawaii to manage other non native and invasive Opuntia sp p. (Moran & Zimmermann 1984). In 1957, C. cactorum was introduced into several Caribbean islands (Nevis, Montserrat and Antigua) to control native as well as nonnative Opuntia spp. (Simmonds & Bennett 1966). Unfortunately, the implementing agencies did not fully consider the potentially injurious environmental impacts of C. cactorum if this insect were to move to neighboring countries where some species of Opuntia are important nat ive species and some are important commercially (Stiling et al. 2004). The first record of C. cactorum in the U S was from Bahia Honda Key, Florida, in October 1989 ( Habeck and Bennett 1990; Dickel 1991). It is uncertain how the moth arrived in Florida, but several interceptions of Caribbean ornamental Opuntia spp. infested with C. cactorum were found at ports of entry in
128 south Florida during the 1980s and 1990s (Pemberton 1995; Zimmermann et al. 2001; Stiling 2002 ; Simonsen et al. 2008). Since its appearance in Florida, C. cactorum has become a threat to native Opuntia spp. in North America. Current management options include the use of Pherocon 1C Wing traps (Trc Incorporated, Salinas, CA) baited with a 3component synthetic sex lur e (Suterra, LLC, Bend, OR) to identify the presence of the moth, coupled with removal of infested plants to reduce C. cactorum populations (Bloem et al. 2005; Hight & Carpenter 2009). Complementary to the detection, monitoring, and removal efforts, implem entation of the Sterile Insect Technique (SIT) is being used to slow the geographic expansion of C. cactorum in the U.S. (Hight et al. 2002; Bloem et al. 2005; Bloem et al. 2007). In Mexico, localized infestations of C. cactorum on two islands of the stat e of Quintana Roo were eradicated in 2008 with an integrated program using pheromone traps, host removal, and the SIT (NAPPO 2006; NAPPO 2008; NAPPO 2009). Bennett and Habeck (1995) suggested biological control as an additional control option that should b e considered for C. cactorum Pemberton and Cordo (2001 a ) reported that s everal larval and pupal parasitoids attacked the cactus moth in South America including species of Hymenoptera ( Braconidae, Chalcidae, and Ichneumonidae), and one Diptera ( Tachinidae ) They also reported on two chalcid species [ Brachymeria ovata (Say) and B. pedalis Cresson] and one unidentified egg parasitoid from the family Trichogrammatidae attacking C. cactorum in Florida. Logarzo et al. (2009) found the larval parasitoid Apant eles alexanderi Brethes (Hymenoptera: Braconidae) and the egg parasitoid
129 Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) attacking C. cactorum in Argentina. Trichogrammatid egg parasitoids have been used successfully for inundative biological control against major lepidopteran pests such as corn borers [ Ostrinia sp. and Diatraea sp. Crambidae] sugarcane borers ( Diatrea spp., Pyralidae), and cotton bollworm ( Helicoverpa spp., Noctuidae) ( Li 1994; van Lenteren 2000). Egg parasitoids are easy to mass rear under laboratory conditions for the application of inundative releases into large areas. Biological control can be used to complement and synergize the application of SIT (Knipling 1992; Gurr & Kvedaras 2010). Studi es have demonstrated that the combination of both techniques is more efficient at controlling pest populations than either technique used by itself (Bloem et al. 1998). Synergistic interactions between the SIT and fruit fly biological control using parasi toids increased the suppression of pest fruit flies, even leading to eradication (Sivinski 1996; Rendon et al. 2006) S IT and biological control have been successfully combined to combat several lepidopteran pests, including C ydia pomonella (L.) (Tortrici dae) (Bloem et al. 1998) and painted apple moth, Orgyia anartoides (Walker) (Lymantriidae) (Suckling et al. 2007) Radiation doses for sterilizing C. cactoblastis adults have been determined to produce partially sterile but fitter males which, when mated with wild females, generate sterile offspring (Carpenter et al. 2005). The combination of egg parasitoid releases and SIT has the advantage that parasitoids manage high pest densities, while SIT works best at low pest densities. In addition, release of s terile insects provides an additional source of
130 eggs for egg parasitoids increasing the ratio of natural enemies to adult hosts. Egg parasitoids and sterile insects are self dispersing and consequently are able to cover wide areas ( Sivinski 1996). Field surveys were conducted in order to identify egg parasitoids already established in North Florida that attack C. cactorum The distribution, seasonality, and parasitism parameters of the Trichogramma species attacking C. cactorum in northern Florida are also reported upon The number of eggs/eggstick between different flight periods and sites was compared to assess host egg resource for egg parasitoids. The effect of C. cactorum eggstick size on level of parasitism was evaluated by comparing number of eggs from parasitized versus unparasitized eggsticks. These data will be beneficial in promoting discussions and action on possible implementation of biological control for the cactus moth and, in particular, assessing the potential of an inundative biological control program against C. cactorum in North America. The use of inundative releases of Trichogramma wasps in combination with SIT will also be discussed. Materials and Methods Field surveys were carried out at six locations (Fig ure 61) in nor th Florida from July 2008 to December 2009. Cactoblastis cactorum adults have three annual flight periods in north Florida (April May, July August, and October November) (Hight et al. 2005; Hight & Carpenter 2009). The s election of study sites was based on existing records of infestations from the literature, personal observations from preliminary surveys and information provided by expert s. Female C. cactorum place their eggs end to end to form a chain that looks like a short stick, and the egg mass is referred to as an eggstick. Although no
131 extensive field surveys were conducted from May to July 2008 at St. Marks and St. George Island, eggsticks with eggs that appeared parasitized were collected and held under laboratory conditions ( 25 1 C 16:8 L:D and 40 60% RH ) until parasitoids emerged. At survey locations, 20 to 30 healthy Opuntia spp. plants were chosen with no to minor feeding damage by cactus moth larvae and an average of at least 50 pads per plant. During weekly visits throughout all three flight periods, any new eggstick was identified by plant, pad, and its general location on the plant so the eggstick could be found during subsequent checks. A mark was made on the plant at the base of the eggsti ck with a felt tip pen and a red tape flag affixed to an insect pin placed near the eggstick to aid in finding the eggstick. The flag was labeled with a unique number to identify each eggstick. The oviposition preferences of C. cactorum females on hos t plants were recorded by classifying the attachment of the eggstick to either a glochid at an areole, to a spine, or on the fruit. Observations on plant habitat and host eggstick distribution within the surveyed site and within the selected plant were co llected to provide additional information on the host finding behavior of egg parasitoids. The number of eggs/eggstick was determined either by a direct count or by a correlated estimate of eggstick length to egg number (2.62 0.013 eggs/mm). The ratio of eggstick length to egg number was calculated in this study by counting the number of eggs in a segment of eggstick, replicated on 20 eggsticks. Eggstick length was estimated in situ by placing a plastic string next
132 to the eggstick and cutting a piece o f equivalent length. The length of the piece of string was then measured to the nearest 0.01 mm with a metric micrometer. Measurements of eggsticks were obtained so that the number of eggs/eggstick could be estimated if the eggstick was lost before it could be collected and directly counted. The fate of each eggstick was determined by making weekly visits to each site to evaluate the status of previously tagged eggstick s. The fate of each eggstick was categorized as follows : eggstick los t; predat ed (vis ible chewing damage) eggs in the eggstick versus non predated eggs; or parasiti zed eggs in the eggstick (black eggs formed before C. cactorum larvae successfully developed) Eggsticks were collected if they were damaged during evaluation or measurement, eggs of the eggstick had hatched, or eggs appeared predated or parasitized. Eggsticks with viable eggs were collected and held in small plastic cups (30 ml) under laboratory conditions ( 25 1 C 16:8 L:D and 40 60% RH ) to record hatch rate. Eggsticks wit h parasitized eggs were collected and monitored in the laboratory to determine the emergence rate, number of eggs/eggstick attacked by parasitoids, number of parasitoids emerging per parasitized egg, and to ascertain the identity of the parasitoids. Paras itoid specimens were submitted to R. Stouthamer, Department of Entomology, University of California, Riverside, for molecular identification. The sequencing of ribosomal DNA Internal Transcribed Spacer 2 (ITS 2) was used to identify the different species of egg parasitoids. Data Analysis The numbers of eggs/eggstick at different flight periods for each surveyed location and the average number of eggs/eggstick at each site were log
133 transformed before analyses to satisfy the assumptions of the analysis of v ariance. Oneway analysis of variance (PROC GLM) was applied to the log transformed data and multiple comparison of means was made with the least statistically difference (LSD) test. Comparison of number of eggs/eggstick that was parasitized versus number of eggs/eggstick not parasitized also was evaluated with a one way analysis of variance (PROC GLM). Because only a few eggsticks with parasitized eggs were collected in this study (see text below), comparisons between eggsticks with parasitized eggs wer e made against the same number of randomly selected eggsticks with unparasitized eggs. Variation between the number of eggs for parasitized eggsticks and the number of eggs for the randomly selected unparasitized eggsticks was analyzed using a folded F test (Davis 2007). Because the variances in numbers of eggs for eggsticks with parasitism and number of eggs in eggsticks without parasitism were not statistically different, means of these two groups were compared with a two sample t test. A Pearsons C orrelation Coefficient (r) was calculated to determine whether the numbers of eggs parasitized were dependent on the number of eggs/eggstick. The SAS Statistical Software Version 9.2 (SAS Institute, Cary, North Carolina) was used to perform the statistic al analyses. Estimates of central tendencies were reported as mean standard error of mean. Results Although host plant species of Opuntia stricta (Haworth) Haworth O. humifusa (Rafinesque) Rafinesque, and O. ficus indica (L.) P. Miller varied among the different geographic regions surveyed; the oviposition preferences of
134 C. cactorum females were similar on the different plant species (Table 61). In this study, parasitized eggsticks of C. cactorum appeared mostly on the areole/glochid structure of the pads (Table 61). In total 1,527 eggsticks with 91,013 C. cactorum eggs, not including 344 eggsticks missing from the field or lost during collection, were tagged on plants of Opuntia spp. (Table 62). Of all the eggsticks checked, 62% were collected on Okaloosa Island and had a mean of 59 1.83 eggs/eggstick. The proportion of eggsticks examined in the laboratory as percentage of all eggsticks surveyed at the six field sites ranged from 53 to 100%, except for 2008 summer at St. George Island and St. Marks National Wildlife Refuge (NWR), in which only 30% and 24% respectively of the monitored eggsticks were examined (Table 6 2). The majority of the eggsticks from these two locations for this flight period were recorded as lost (Table 62). The cause for this high number of lost eggsticks is not clear. Several biotic and abiotic factors could have contributed to the high number of lost eggsticks. During summer 2008, 23% of eggsticks examined from St. Marks had eggs that were preyed upon compared with less than 3% in other locations. Although not directly observed at St. George Island or St. Marks, substantial predation of C. cactorum eggs by ants has been recorded in South Africa (Robertson 1984). Because the plants sur veyed at St. Marks were located within 100 m of the waters of the Gulf of Mexico, strong winds characteristic of coastal regions could have knocked eggsticks off the plants. All other study sites were along the Gulf Coast; in none of them were the plants as close to the water as at St. Marks. In addition, heavy rainfall may have separated the eggsticks from plants, but we do
135 not have any data on the severity of the rain storms at different study sites. Cactoblastis cactorum life table studies in Argentin a (Logarzo et al. 2009) and South Africa (Robertson & Hoffmann 1989) identified rain and wind as major factors contributing to mortality of eggs. Surveyed sites and oviposition periods were analyzed to evaluate their influence on number of eggs/eggstick. Eggsticks were collected for multiple oviposition periods at three sites (St. George Island, St. Marks, and Okaloosa Island) (Table 62). The numbers of eggs/eggsticks for the different oviposition periods were not statiscally different for St. George I sland (F = 1.84, df = 1, P = 0.18), St. Marks (F = 93.86, df = 3, P = 0.07), or Okaloosa Island (F = 0.22, df = 3, P = 0.88). Because the numbers of eggs/eggstick for multiple oviposition periods were not different, eggsticks from all flight periods were pooled to calculate the means for those sites [St. George Island (62 2.8), St. Marks (53 2.8), and Okaloosa Island (59 1.8)]. The pooled eggsticks were used to compare the number of eggs/eggstick between all six sites and differences were indeed fou nd ( F = 11.44, df = 5, P < 0.0001) (Table 6 2). Female C. cactorum laid similar numbers of eggs/eggstick for each of the three oviposition periods but not at all six survey sites along the Florida panhandle. The longest eggsticks were observed at St. George Island, Pensacola Beach, and Okaloosa Island (Table 62). Significantly smaller eggsticks were recorded at St. Marks and Mexico Beach (Table 62). Panacea had significantly smaller number of eggs/eggstick than all other sites (Table 62). The cause of differences between eggsticks at the various sites was unclear.
136 Studies in South Africa identified differences in total fecundity of C. cactorum due to host plant species, the flight period when eggs were laid, and the temperature during oviposition ( Robertson 1989). We did not distinguish eggsticks collected from different host plants (Table 61). While South African female C. cactorum had significantly higher fecundity during the summer flight (Robertson 1989), this study did not show any differenc e in number eggs/eggstick between flight periods in North Florida. Cactoblastis cactorum has a tendency to oviposit on plants with high nitrogen (Myers et al. 1981; Robertson 1987), but we have no direct measurements of plant quality at our sites. Compa ring the number of eggs/eggstick for eggsticks that were parasitized (38 13.7) (Table 63) against unparasitized eggsticks (61 13.1) revealed there was a difference (pooled t test = 3.14, df = 12, P = 0.0085). Although the number of eggs/eggstick was highly variable, the variation of the number of eggs/eggstick for parasitized versus the randomly selected unparasitized group was similar (folded F test = 1.10, df = 6, P = 0.91), suggesting that the difference found between the two groups was not driven by unequal or extreme variation. However, there was not a significant correlation between the number of eggs/eggstick and number of eggs parasitized by Trichogramma spp. (N = 7, r = 0.16, P = 0.74). Therefore, whereas female Trichogramma spp. parasiti zed eggsticks with small number of eggs, they did not parasitize more eggs as the number of eggs in an eggstick increased. The average number of eggs parasitized in an eggstick was 9 ( 5.8).
137 Ten eggsticks were found parasitized at three of the six sites surveyed (Pensacola Beach, St. Marks, and Okaloosa Island). Five of the parasitized eggsticks were found at Okaloosa Island. Parasitized eggsticks were found during all three oviposition periods of C. cactorum females: the spring flight (St. Marks and Pensacola Beach), summer flight (Pensacola Beach), and fall flight (Okaloosa Island). Of the 496 eggs in the ten parasitized eggsticks, a total of 89 eggs (or 18%) were parasitized, resulting in the emergence of 181 adult parasitoids with a sex ratio of 70% ( 14) females (Table 63). The level of parasitism by Trichogramma spp., relative to the total number of eggs examined during the different flight periods for each site, was less than 0.2% of total C. cactorum eggs collected (Table 63). We did not observe any parasitized eggsticks at St George Island, Mexico Beach, or Panacea. Two species of Trichogramma were reared from C. cactorum eggsticks in North Florida (Table 63) and identified by differences in IST2 sequences. Trichogramma pretiosum Riley was collected at St. Marks, Pensacola Beach, and Okaloosa Island, whereas T. fuentesi Torre was recovered only from Okaloosa Island. It was not possible to identify two collections of Trichogramma spp. from Okaloos a Island for two reasons: (1) because a good molecular sequence could not be obtained and (2) the sequence was not in the database and possibly represents a new species in the T. pretiosum group (R. Stouthamer, UC Riverside, personnel communication). Dis cussion More than 15 million ha of agriculture and forestry worldwide are treated annually with Trichogramma egg parasitoids (van Lenteren 2000).
138 Trichogrammatid wasps have been used successfully in inundative release programs against lepidopteran pests i n greenhouses and crop production worldwide (Smith 1996). Inundative releases of Trichogramma spp. have been implemented in Florida to control major lepidopteran pests of collards, cabbage, soybeans, bell peppers, tomatoes, corn, and tobacco (Martin et al 1976). Trichogramma pretiosum is commonly found in the Western Hemisphere. This species has been released commercially against major lepidopteran pests such as cotton leafworm [ Alabama argillacea (Hbner) Noctuidae] corn earworm [ Helicoverpa zea (Bod die), Noctuidae], tomato pinworm [ Keiferia lycopersicella (Walshingham ), Gelechiidae], sugarcane borers ( Diatraea spp.) (Crambidae) and cabbage looper [ Trichoplusia ni (Hbner ), Noctuidae] ( Pinto et al. 1986; Hassan 1993; Li 1994; Monje et al. 1999) Tri chogramma fuentesi ha s been recorded in countries in South America (Argentina, Columbia, Mexico, Peru, and Venezuela) and in the U.S. (Alabama, California, Florida, Louisiana, New Jersey, South Carolina and Texas) (Fry 1989, Pinto 1999). Its primary hosts are species from the family Noctuidae such as H. zea and Heliothis virescens (F.) and from the family Pyralidae such as Diatrea saccharalis (F.), Ephestia kuehniella Zeller, and Ostrinia nubilalis (Hbner ) ( Fry 1989; Wilson & Durant 1991; Pintureau et al. 1999; Querino & Zucchi 2003). They are also widely used for pest control in orchards (Olkowski & Zang 1990) The observed low incidence of the wasps in natural areas might be explained by unfavorable environmental factors or natural plant chemicals (Smith 1996; Romeis et al. 1997, 1999). However, contrary to
139 other natural enemies, Trichogramma can be easily and cheaply mass reared for the implementation of an augmentative biological control program. The potential for inund ative releases of Trichogramma spp. as a strategy against C. cactorum is currently being investigated using laboratory colony of T. fuentesi originating from insects reared from parasitized C. cactorum eggsticks. Biological characteristics (sex ratio, egg load, and longevity) and different behavioral mechanisms (influence of parasitoid age, density, and host age on parasitism) involved in host finding of T. fuentesi reared on C. cactorum eggs are being evaluated. Inundative releases of T fuentesi could be integrated into the current pest management strategy that is based on SIT and removal of infested hosts during the three flight periods of C. cactorum The present field survey was useful in identifying potential biological control agents that might be i ntegrated in a combined pest management strategy against C. cactorum
140 T able 61 Sites surveyed in North Florida for egg parasitoids of C actoblastis cactorum eggsticks on O puntia sp p and additional information on moth oviposition preference. Site GPS c oordinate Dates eggsticks surveyed Total number s urveys Species of Opuntia host p lant Number h ost plant examined Total number eggsticks evaluated Percent e ggstick s at a ttachment locationa Areole/ Glochid Spine Fruit Missing b Pensacola Beach N30.33083 W87.15869 Summer 2008 (Jul. 10-Sep. 10, 08) 10 O. stricta O. humifusa O. ficus indica 20 120 50 34 16 0 St George Island N29. 39051 W84. 53577 Summer 2008 (Jul.17-Sep.19, 08) 10 O .stricta 23 105 63 30 7 0 Fall 2008 (Sep. 25, 08Feb. 25, 09) 20 27 28 89 11 0 0 St Marks (NWR) N30.07772 W84.18242 Summer 2008 (Jul. 15, -Sep. 12, 08) 18 O. stricta O. humifusa 30 45 80 13 7 0 Fall 2008 (Oct. 01, 08-Feb. 25, 09) 20 30 9 78 0 22 0 S pring 2009 (Apr. 17Jul. 15, 09) 13 35 47 88 2 0 10 Fall 2009 (Oct. 07, 09Jan. 12, 10) 35 151 n/a n/a n/a n/a a Attachment locations of eggsticks that were not determined is indicated by n/a. b Information about eggstick attachment failed to be recorded.
141 Table 61. Continued Site GPS c oordinate Dates eggsticks surveyed Total number s urvey s Species of Opuntia h ost p lant Number h ost p lant e xamined Total number eggsticks evaluated Percent e ggstick at a ttachment locationa Areole/ Glochid Spine Fruit Missing b Mexico Beach N29. 56969 W85. 25250 Fall 2009 (Oct. 21 -Nov.12, 09) 3 O. ficus indica n/a 29 n/a n/a n/a n/a Panacea N30.0 995 W84. 22086 Fall 2009 (Oct. 21 -Nov.12, 09) 3 O. stricta O. ficus -indica n/a 65 n/a n/a n/a n/a Okaloosa Island N30. 39845 W86. 59304 Fall 2008 (Oct. 08, 08-Feb. 27, 09) 21 O. ficus indica 10 186 81 18.5 0.5 0 Spring 2009 (Apr. 08 Jul.08, 09) 14 18 308 79 15 1 4 Summer 2009 (Jul. 01-Sep. 25, 09) 18 18 280 77 19 2 2 Fall 2009 (Sep. 18, 09Jan. 12, 10) 25 151 n/a n/a n/a n/a a Attachment locations of eggsticks that were not determined is indicated by n/a. b Information about eggstick attachment failed to be recorded.
142 Table 62. Number of Cactoblastis cactorum eggsticks collected, lost in the field, examined in the laboratory, and number of eggs per eggstick SE at different sites in North Florida for different oviposition periods. Site Flight p eriod Total number e ggsticks t agged Total number ( percent ) e ggsticks l ost Percent e ggsticks e xamined Total number m oth e ggs e xamined Mean number e ggs/ e ggstick SE Overall mean eggs/eggstick SE at each sitea Pensacola Beach Summer 2008 1 20 69 (58) 42 7, 402 62 1. 5 621.5 a St George Island Summer 2008 105 84 (70) 30 6,685 64 2.0 622.8 a Fall 2008 28 13 (46) 53 1,614 58 3.6 St Marks (NWR) Summer 2008 45 35 (77) 24 3 ,088 6 8 9 532.8 b Fall 2008 9 4 (44) 55 513 57 3.3 Spring 2009 47 2 3 (46) 54 2,561 54 3. 1 Fall 2009 151 0 (0) 100 6,917 46 1. 4 Mexico Beach Fall 2009 29 0 (0) 100 1,522 52 0 523.0 b Panacea Fall 2009 65 0 (0) 100 2,892 45 1. 9 451.9 c Okaloosa Island Fall 2008 186 61 (29) 71 11 118 60 1. 4 591.3 a Spring 2009 308 3 (1) 99 20 527 61 1.2 Summer 2009 28 0 21 (8) 92 17 126 60 1. 1 Fall 2009 151 1 (0.6) 99 8 638 57 1.2 a Means with different letter are statistically different (P < 0.05).
143 Table 63. Location and date parasitized Cactoblastis cactorum eggstick was collected, identity of parasitoid species, number of eggs per eggstick, number of parasitized eggs, number of parasitoids emerged, female ratio, and parasitism level of egg parasitoids attacking C. cactorum in North Florida. Site Collection d ate Flight p eriod Trichogramma sp. Number e ggs/ e ggstick Number p arasitized e ggs/ e ggstick # (%) number p arasitoids e merged Percent f emales Level of p arasitism (%) St Marks 05/15/08 Spring 08 T. pretiosum 73 5 (7) 8 75 n/a a 05/15/08 T. pretiosum 78 17 (22) 34 85 Pensacola Beach 04/22/08 Spring 08 Trichogramma sp. 88 19 (22) 18 77 n/a 08/06/08 Summer 08 T. pretiosum 44 8 (18) 5 40 0.2 08/13/08 T. pretiosum 18 6 (33) 10 70 Okaloosa Island 10/16/08 Fall 08 T. fuentesi 20 2 (10) 11 73 0.1 10/16/08 T. pretiosum 42 10 (24) 7 71 10/16/08 T. fuentesi 56 6 (11) 16 62 11/03/08 Trichogramma sp. 52 3 (6) 9 56 10/23/09 Fall 09 T. fuentesi 25 13 (52) 63 89 0.1 a Indicates that the level of parasitism was not determined.
144 Figure 6 1. Locations survey ed for egg parasitoids of Cactoblastis cactorum in North Florida. Pensacola Beach N30.33083 ; W87. 15869 Mexico Beach N29. 56969 ; W85. 25250 Okaloosa Island N30. 39845 ; W86. 59304 St. Marks (NWR) N30.07772; W84.18242 Panacea N30.0 995; W84.22086 St. George Island N29. 39051 ; W84. 53577
145 Figure 62. Ms. Paraiso surveying cactus plant for Cactoblastis c actorum eggsticks. Figure 63. Parasitized Cactoblastis cactorum egg in an eggstick on Opuntia cactus pad. Photo S. Hight Photo S. Hight
146 CHAPTER 7 TRICHOGRAMMA FUENTESI A NEWLY DISCOVERED POTENTIAL BIOLOGICAL CONTROL AGENT OF CACTOBLASTIS CACTORUM EVALUATION OF BIOLOGICAL PARAMETERS Previous surveys for natural enemies of C. cactorum in North Florida led to the discovery of T. pretiosum (Riley) and T. fuentesi Torre (Paraiso et al. 2011; Chapter 6). Inundative releases of several Trichogramma spp. such as T. exiguum Pinto & Platner against heliothine pests of cotton (Suh et al. 2000) or against the Nantucket pine tip moth [ Rhyacionia frustana (Comstock)] in Virginia pine (Philip et al. 2005), have failed to provide an adequate level of pest suppres sion. Therefore, prior to field release it is important to undertake detailed studies of biological and ecological characteristics of prospective agents before releas e in the field (Ashley et al 1974; van Lenteren et al. 2003; Dannon et al. 2010) Some of the important characteristics that need to be studied include: sex ratio, longevity, influence of parasitoid age, and host age on percent parasitism. Trichogramma wasps require a source of carbohydrate to maintain basic physiological activities (Romeis et al. 2005). Generally, raisins and pure or diluted honey are used in experimental studies as a source of food to maintain Trichogramma colonies (Morrison 1985). The presence and nutritional quality of a supplemental source of food have been reported to increase the longevity of Trichogramma spp. (Laetimia et al. 1995; Oliveira et al. 2003). In addition, a source of carbohydrate can, in some cases, influence fecundity and egg resorption in parasitoids (Heimpel et al. 1997). Female parasitoid age is another biological characteristic that can affect the success of an augmentative biological control agent (Amalin et al. 2005). Studies have shown that trichogrammatids under or over a certain age are not able to parasitize their hosts
147 (Rajapakse 1992; Amalin et al. 2005). Host age also impacts the level of parasitism because trichogrammatid parasitoids favor young host eggs (Hagvar & Hofsvary 1986; Sequeira & Mackauer 1988; Amalin et al. 2005) and younger host age has an influence on parasitoid o ffspring fitness (Sequeira & Mackauer 1992, 1994). In the present study, we examined the influence of three types of supplemental food sources on the longevity of T. fuentesi in the laboratory. In addition, optimum oviposition age of female T. fuentesi was assessed in order to obtain information for optimizing possible inundative releases against C. cactorum T his chapter also provides information on host age on parasitism rates, number of parasitoid progeny produced (fecundity), number of parasitoids e merged/parasitized egg (fertility), and sex ratio. Materials and Methods Rearing Procedures and General Methods Experiments were conducted at the USDA, Agricultural Research Service and the Florida A&M University/Center for Biological Control L aboratory Tallahassee, FL. Trichogramma fuentesi females used in the study were obtained from a colony established from field collected material. Species identity was confirmed by DNA ITS 2 sequences performed by Dr. R. Stouthamer (Department of Entomology, University of California, Riverside CA). Cactoblastis cactorum eggs derived from a colony maintained on artificial diet were used as hosts during parasitoid rearing procedures and for the experiments. To culture T. fuentesi host eggsticks were glued on note card strips (4 x 2 cm) with nontoxic Elmers glue (E l mers Products, Inc. Columbus, OH) (Figure 7 1) The note card strips were then placed into standard plastic petri dishes (9 x 2 cm) lined with filter paper. A fresh raisin was glued on a 1 x 1 cm note card in the
148 center of the p etri dish to provide a supplemental source of food for emerging wasps (Figure 7 2) Petri dishes were sealed with Parafilm (Pechiney P lastic Packaging, Menasha, WI) and arranged on plastic trays lined with moist wipes to maintain relative humidity at 60 80% (Figure 7 3) The cultures were maintained in a growth chamber at 28 1C, 16:8 L:D. Additional growth chambers maintained at 25 1C, 16:8 L:D and 60 80% RH were used for incubation of experimental units in all experiments. Effect of Presence and Type of Diet on Female Parasitoid Longevity We tested the effect of no supplemental source of food and presence of pure honey or raisins on the longevity of female parasitoids. Individual newly emerged (0 24 h old) T. fuentesi females were collected from the rearing colony and placed in a plastic p etri dish (3 5 x 1 c m) lined with filter paper A source of energy consisting of a drop of pure honey or a raisin was added to the center of the container A set of control petri dishes did not have any supplemental source of food. Petri dish es were sealed with Parafilm and arranged on a plastic tray lined with moist paper wipes to maintain the relative humidity The containers were incubated until all females died. The experiment was replicated 20 times Influence of T. fuentesi Female Age on Percent Parasitism The effect of female age on the percentage of host eggs parasitized was tested for females ranging from one to five days old. Oneday old female parasitoids were isolated from the rearing colony and individually transferred to a p etri dish (3.5 x 1 cm) lined with filter paper as described above. The experimental arena did not have a source of food for the wasps. Females were exposed to 60, two day old, C. cactorum eggs. Petri dishes were sealed with Parafilm and arranged on a plastic tray lined with moist paper wipes to maintain the relative humidity and then incubated. E ggsticks were
149 replaced daily until the females were five day s old (a dult female parasitoids without any food live an average of four days data from Experiment 1) The experiment was replicated 20 times The number of parasitized eggs was determined on a daily basis Effect of Female Mating Status on Percent Parasitism Newly emerged (0 24 h) females and males were collected from the rearing colony and stored in a petri dish for 24h to allow them to mate. Female wasps were distinguished from male based on antennae characteristics. Female Trichogramma have clubbed antennae with few short hairs on segmented flagellum while males have antennae with unsegmented flagellum and long hairs (Flanders 1965). Individual mated females were then transferred as above, into a p etri dish (3 .5 x 1 cm) lined with filter paper. Single f emale parasitoids were exposed to 60, two day old C. cactorum eggs for 24 h The Petri dishes were sealed with Parafilm and incubated. Female parasitoids and eggsticks were replaced daily The experiment was replicated 20 times and t he number of parasitized eggs and the percent parasitism were record ed Rates of parasitism for these mated females were compared against unmated, 24h females used in Experiment 2. Influence of Host Age on Pe rcent Parasitism Cactoblastis cactorum females lay an average of 70 90 eggs/eggstick (Zimmermann & Prez Sandi 2006). Eggs hatch in approximately 3 weeks (Zimmermann et al. 2001). A no choice experimental design was used to assess the influence of t en different C. cactorum host age groups (1, 6, 7, 9, 11, 12, 13, 14, 15, and 20day old) on percent parasitism. Threeday old, randomly chosen, honey fed, mated female T. fuentesi were isolated from the rearing colony and each placed in an individual Pet ri dish (3 5 x 1 cm) lined with filter paper In total, 60 host eggs belonging
150 to one of the different age groups were placed in the center of the Petri dishes E ggsticks were removed after 48 h and individually transferred in to plastic cup s ( 30 ml ) for 10 days to allow all parasitoids to emerge. The experiment was replicated 20 times and the percent parasitism total number of emerged parasitoids, number of emerged parasitoids per parasitized eggs, and sex ratio were determined. Statistical Analysis Dat a collected were subjected to analysis of variance ( PROC ANOVA & PROC GLM ). The SAS Statistical Software Version 9.2 (SAS Institute, Cary, NC) was used to perform the statistical analyses. The effect of female age on the number of eggs parasitized and p ercent of parasitism between females that did or did not have mating experience was tested with a general linear model ANOVA followed by Tukey s test for the separation of means (PROC ANOVA). In addition, the effect of host egg age on percent level of parasitism, number of progeny, number of successfully emerged eggs, and percentage of females produced was analyzed by polynomial regression. Results Effect of Presence and Type of Diet on Female Longevity The provision of a food supplement had a significant influence on the survival of female parasitoids. Without a supplemental food source, females survived for an average of 4 0.58 days (F = 23.14, df = 1, 12, P < 0.001, R2 = 0.85), whereas with a raisin or honey supplement, females lived for 8 0.07 days (F = 595.03, df = 1, 12, P < 0.001, R2 = 0.99) and 11 0.79 days (F = 670.73, df = 1, 12, P < 0.001, R2 = 0.99), respectively. Statistical analysis of the data showed a difference in the number of days parasitoids survived between honey and raisin fed females (F = 43.20, df = 2, 57, P <
151 0.001, R2 = 0.60). The maximum survival time was 16 days for a female which was fed honey. Thus, the type of food also affected the longevity of the fem ale parasitoids. Influence of Female Parasitoid Age and Mating Status on Percent Parasitism Percent parasitism varied significantly with female age (F = 15.42, df = 4, 190, P < 0.0001) (Table 71). The number of parasitized eggs/eggstick was the highest for one and twoday old female parasitoids and this declined rapidly thereafter (Table 71). In addition, the results showed that parasitism increased when females were mated prior to exposure to host (F = 14.78, df = 1, 190, P < 0.001). Unmated females had none to about 20% of parasitism after the second day. In contrast, mated females were still parasitizing cactus moth eggs after four days (Table 71). Influence of Host A ge on Percent Parasitism The number of parasitized eggs and the number of emerged parasitoids/eggstick declined with host egg age. (F = 19.53, df = 3, 196, P < 0.0001 ; and F = 16.53, df = 3, 196, P < 0.0001, respectively ) (Table 72) The relationship between host egg age and the number of eggs parasitized was best represented by a cubic polynomial equation (y = 4.81 1.02 x + 0.074 x2 0.0018x3, R2 = 0.23). Trichogramma fuentesi displayed a preference for one day old host eggs; t he highest level of parasitism and number of progeny produced was recorded for oneday old host eggs. F emale parasitoids did not produce progeny in host eggs older than 14 days (Table 72) A cubic polynomial equation also described the relationship betw een egg age and the number of parasitoids emerged (y = 16.17 3.56x + 0.27 x2 0.0065x3, R2 = 0.20). However, host egg age did not have a significant influence on the sex ratio or on the total number of parasitoids emerging / parasitized egg (Table 7 2) Although polynomial regression revealed significant relationships between host egg age and the number of parasitized
152 eggs and the number of parasitoids/eggstick, only about 20% of the variation in the data was explained by our models (Figure 74) The hig h significance level observed indicates the influence of host egg age, but additional factors are important in explaining the variation in our data. Discussion This study provides information on the biological parameters of T. fuentesi a naturally occur ring biological control agent that might have potential application in augmentative releases against C. cactorum Although there is a considerable amount of information on the biology of some Trichogramma spp., little is known about this particular species. Trichogramma fuentesi has been recorded in the neotropics, including Argentina, Colombia, Mexico, Peru, and Venezuela and in several U.S. states (Alabama, California, Florida, Louisiana, New Jersey, South Carolina and Texas) (Fry 1989; Pinto 1999). It s primary hosts are mainly noctuid species (Fry 1989; Wilson & Durant 1991; Pintureau et al. 1999; Querino & Zucchi 2003). Trichogramma spp. are generally proovigenic parasitoids (Pak & Oatman 1982; Fleury & Boultreau 1993). However, several studies hav e suggested that Trichogramma have synovigenic tendencies, as many species develop additional eggs as female age increases (Houseweart et al. 1983, Bai & Smith 1993, Mills & Kuhlmann 2000). Trichogramma wasps require sugar as a source of energy to sustai n major physiologic al processes (Romeis et al. 2005). In nature, Trichogrammatids may obtain sugar from floral nectar, extrafloral nectar ies honeydew, and plant sap (Wackers 2005). F ood deprived parasitoids will first search for food resources before they search for hosts (Hegazi et al. 2000). In addition, sugar intake influences overall individual flight and foraging behavior ( Forss e et al 1992; Pompanon et al. 1999; Romeis et al. 2005).
1 53 Hegazi et al. (2000) observed that honey deprived T. cacoeciae Marchal only attacked one host patch whereas fed females were more likely to move to a second host egg patch. Other studies report significant increas e s i n the longevity of Trichogramma spp. adults when they are provided with a supplemental food source [e.g., T. minutum (Riley) (Laetemia et al. 1995); T. galloi Zucchi (Oliviera et al. 2003); and T. pretiosum (Berti & Marcano 1993)] whereas others reported no differences in longevity if adults were or were not fed [ T. demoraesi Nagaraja (Oliviera et al. 2003)]. Female T. fuentesi without supplemental food never lived more than 5 days. Preferences for sucrose over fructose and glucose have been shown for Hymenoptera (Fonta et al. 1985; Cornelius et al. 1996; Koptur & Truong 1998). The main sugars in ho ney are the monosaccharides fructose and glucose. Additionally, various oligosaccharides such as sucrose, maltose, trehalose and turanose have also been detected (Siddiqui 1970; Doner 1977). Honey also contains proteins, enzymes and amino acids (Vela et al. 2007). As in honey, the main sugars in raisins are fructose and glucose with minimal amount of sucrose. Raisins are a good source of vitamins, minerals and phytochemicals (Williamson & Carughi 2010). In the present study, we examined the influence o f the presence and the type of food source on the longevity of T. fuentesi females. We observed that a supplemental source of food prolonged the longevity of T. fuentesi by an average of 11 days when given honey and 8 days when given a raisin Although a comparison of the nutritional quality of food sources was not done, the observed differences in longevity may be explained by the readily accessible carbohydrate in honey as compared to the raisin diet.
154 Various studies have shown that the parasitoid age also can affect the level of parasitism at the time of release (Hentz 1998; Honda & Kainoh 1998). Knowledge of optimal ovipositional time for T. fuentesi females provides information on when they are the most fecund This information is important in deciding at what age parasitoids should be release d in the field in order to obtain a higher levels of parasitism (Amalin et al. 2005). In this study, o ne to two day old mated female exhibited the highest level of parasitism (Table 71) Th is level of parasit ism decreased from the third day until the death of the parasitoids. The highest level of parasitism was observed in oneday old unmated parasitoids and one to threeday old mated females (Table 71) Gregarious egg parasitoids, such as Trichogramma spp. can mate locally at emergence or disperse and mate later with a nonlocal mate (Martel & Boivin 2004). Hardy et al. (2007) reported studies showing that mating did not affect oviposition behavior of parasitoids wasps. Other studies have shown differenc es in oviposition behavior between virgin and mated females. Mated females oviposited immediately while unmated females laid few eggs in hosts and consequently mated with their sons (van den Assem & Visser 1976; van den Assem et al. 1982). Studies also have shown that although mating resulted in a larger number of progeny (Tagawa et al. 1987), it did not affect the number of parasitized hosts (King et al. 2000). In this study, percent parasitism did not significantly increase with mating except for twod ay old females (Table 71). Therefore, inundative releases of twoday old mated T. fuentesi females should be used against C. cactorum eggs to increase control levels. Younger and older C. cactorum eggs coexist in the field during each flight period. T richogramma spp. generally prefer younger host eggs (Pak 1986; Godin & Boivin
155 2000; Takada et al. 2000). For instance, T. dendrolimi Matsumura females spent a longer time on younger host eggs (from oneto threedays old) and did not parasitize eggs older than four days (Takada et al. 2000). In addition, the number of progeny decreased significantly with older host eggs (Hiehata et al. 1976; Hinz & Andow 1990; Miura & Kobayashi 1998; Takada et al. 2000). In this study, the highest level of parasitism and number of parasitoid progeny was observed on oneday old host eggs (Table 72). The number of eggs parasitized decreased as the host eggs age increased. Trichogramma fuentesi females did not parasitize C. cactorum eggs older than 13days old (Figure 71) Older host eggs have been shown to be less suitable for Trichogramma egg development due to depletion of essential nutrients (Ruberson & Kring 1993). Trichogramma spp. are generally unable to develop from older eggs, usually because of host embryo rotation or cephalic capsule sclerotization (Guang & Oloo 1990) Therefore, young host eggs are preferantially selected for oviposition ( Hagvar & Hofsvary 1986; Sequeira & Mackauer 1988; G odin & Boivin 2000). Female parasitoids generally lay a higher percentage of females in younger host eggs presumably due to their higher fitness. In this study, although sex allocation was not influenced by host age, femalebiaised progeny were recovered in all experimental treatments (Table 72). In summary, o ur results showed that a honey supplement must be provided in mass rea r ing programs to establish a s ustainable production system of population of T. fuentesi They also showed that one to two day old mated T. fuentesi females should be released in the field to obtain a significant level of parasitism. Additionally, T fuentesi displayed a preference for younger host eggs and t herefore inundative
156 releases should be timed to coincide with the beg inning of the C. cactorum oviposition period or frequent releases made to attack newly layed C. cactorum eggs.
157 Table 71. The influence of age and mating status on number of Cactoblastis cactorum eggs parasitized by Trichogramma fuentesi F emale age (Days) n = 20 Mean # of e ggs p arasitized ( S.E.)* Mated Unmated 1 5.35 0.97 a A 4.00 1.06 a A 2 5.90 1.39 ab A 1.75 0.78 ab B 3 2.80 0.75 bc A 0.05 0.05 b A 4 0.85 0.65 c A 0.00 0.00 b A 5 0.00 0.00 c A 0.00 0.00 b A *Means with different lower case letter in columns or capital letter in rows are statistically different according to Tukeys Least Squared Means Comparison test at P 0.05.
158 Table 72. Influence of Cactoblastis cactorum egg age on parasitization by Trichogramma fuentesi Age of C. cactorum (days) Mean # Egg age Eggs parasitized Emerged parasitoids/eggstick Emerged parasitoids/parasitized egg % Females 1 3.95 1.28 a 13.20 4.70 a 3.42 0.80 60 14 6 0.50 0.25 b 2.00 0.89 b 5.53 1.34 98 2 7 0.65 0.43 b 1.00 0.55 b 2.00 0.58 50 29 9 0.80 0.34 b 2.85 1.43 b 3.32 0.75 50 0 11 0.30 0.21 b 1.40 0.82 b 5.83 1.17 81 9 12 0.30 0.15 b 1.55 0.87 b 6.50 2.73 71 5 13 0.20 0.20 b 0.65 0.65 b 3.25 0.00 85 0 14 0 0 0 0 15 0 0 0 0 20 0 0 0 0 *Means ( S.E.) with different letter are significant according to Tukeys Least Squared Means Comparison test at P Because no significant relationship was found between age of host egg and number of parasitoids per parasitized eggstick or percent females, means were not separated.
159 Figure 71. Nonparasitized (left) Cact oblastis cactorum eggsticks glued on paper strip nex t to glued parasitized eggs (right) in eggsticks by Trichogramma fuentes i. Figure 72 Rearing settings for Trichogramma fuentesi using Cactoblastis cactorum as host eggs
160 Figure 7 3. Arrangement used to increase the relative humidity in rearing cultures of Trichogramma fuentesi
161 Figure 7 4. A regression showing the influence of Cactoblastis cactorum host egg age (1 20 days) on level of parasitism by T richogramma fuentesi
162 CHAPTER 8 TRICHOGRAMMA FUENTESI EVALUATION OF FUNCTIONAL AND NUMERICAL RESPONSE Development of an organism for augmentative biological control is usually preceded by studies to assess the biocontrol agents potenti al for success. Such studies typically include an assessment of critical biological and behavioral criteria, such as dispersal ability, reproductive rate, longevity and climatic tolerance (Kalyebi et al. 2005) Other parameters often measured are the f u nctional and numerical responses, which provide information on natural enemy searching ability and attack rate efficiency (van Lenteren & Bakker 1976; Hassell 1978, 2000; Pandey et al. 1984; Houck & Strauss 1985; Walde & Murdoch 1988; Hugues et al. 1992; W iedenmann & Smith 1993; Bernal et al. 1994; Kumar et al. 1994; van Alebeek et al. 1996; Berryman 1999; Fernandez Arhex & Corley 2003). Functional response was first defined by Solom o n (1949) as the relationship between the numbers of prey consumed by a pr edator with increasing prey density. Holling (1959) described three types of functional response: t ype I (linear), type II (hyperbolic), and type III (sigmoid). In type I functional response, the number of prey consumed increases linearly with host density In a type I I functional response, the number of prey attacked increase s with increasing prey density and then decreases at higher host densities reaching a plateau due to prey handling time At low prey density, a type III functional response has a similar increase in prey consumption as in type II, however attack rate decreases due to increase d searching activity (Hassell 1978). Numerical response is the other component of the host parasitoid complex dynamic. It is described as the number of parasitoid offspring produced per host killed
163 (Ives & Settle 1996). In other words, the numerical response is an increase in reproduction rate due to changes in host density (Solomon 1949). Trichogramma wasps usually display type I or type II f unctional responses under laboratory conditions (Smith 1996). In the context of implementing an effec tive inundative egg parasitoid release program, the use of species with a type II or type III functional response has been recommended (Laumann et al. 200 8) Female parasitoids also react to increasing numbers of host eggs by increasing the number of progeny laid. Increase in parasitoid progeny contributes to the stability of the biological control agent population. This change in ovipositional behavior is defined as a numerical response (Podoler et al. 1978). Against this background, the functional response of T. fuentesi was examined to ascertain whether this species can be used in inundative program against C. cactorum Materials and Methods Rearing P rocedures Experiments were conducted at the USDA, Agricultural Research Service and Florida A&M University, Center for Biological Control L aboratory Tallahassee, FL. Trichogramma fuentesi females used in this study were isolated from a colony or iginating from field collected material The identities of the wasp species w ere confirmed by R. Stouthamer ( Department of Entomology, University of California, Riverside, CA) by differentiating DNA ITS 2 sequences Eggs from C cactorum cultures maintai ne d on an artificial diet were used as hosts for parasitoid rearing and as the source of experimental eggs To culture T. fuentesi h ost eggsticks were glued on note card strips (4 x 2 cm) with nontoxic Elmers glue. The note card strips were then plac ed in to standard plastic petri dishes (9 x 2 cm) lined with filter paper. A fresh raisin
164 w as glued on a 1 x 1 cm note card in the center of the petri dish to provide a source of energy to emerg ing wasps. Newly emerged parasitoids were held together, with food but without hosts, for 3 days before conducting experiments to allow mating and feeding. Based on previous studies, T fuentesi reared on C. cactoblastis eggs achieve their optimal oviposition activity 2 to 3 days after emer gence (Chapter 7). Petri dishes were sealed with Parafilm (Pechiney Plastic Packaging, Menasha, WI) and arranged on plastic trays lined with moist wipes to increase relative humidity at 60 80%. The cultures were maintained in growth chamber at 2 8 1C, 16:8 L:D. Functional and Numerical R esponse Experiments The functional response of T. fuentesi was studied using twoday old C. cactorum eggs at six different treatment densities (10, 20, 40, 60, 80, and 100 eggs ). Preliminary experiments using various arena sizes (gelatin capsule size 0, glass vial 5 x 1.5 cm, plastic petri dish 3 5 x 1 cm and plastic petri dish 9 x 2 c m) showed that the smaller size of petri dish (3 5 x 1 cm) reduced the variability in parasitoid attack rate. For each treatment a single eggstick was placed in an experimental arena lined with a filter paper (Figure 8 1). To assess the appropriate exposure time, single T. fuentesi females were confined with C. cactorum eggs for 6, 12, 24, and 48 h. A female parasitoid in contact with host eggs for 48 h exhibited the most consistent attack rate. A single threeday old mated and fed female parasitoid was isolated from the adult colony and placed in the experimental arena for 48 h. Female parasitoids had no experience with host eggs before they were used in this experiment. Experimental eggsticks were recovered after 48h and placed in a small plastic cup (30 ml) for incubation. Cups were kept in growth chamber s ( 2 8 1 C, 16:8 L:D and 60 80% RH) until parasitoid emergence or for a maximum of ten days. The experiment was replicated 20 times for each treatment The
165 following d ata were recorded: T he proportion of eggsticks parasitized the number of parasitized eggs, and the sex ratio for emerged parasitoids. The numerical response for T. fuentesi was determined by recording the number of emerged parasitoids and the number of emerged parasitoids per parasitized host egg for all treatments mentioned above. Statis tical A nalysis Data collected were analyzed using general linear models with number of host eggs as the source of variation (PROC ANOVA & PROC GLM) (SAS Institute 19 9 9). The proportion of eggsticks parasitized, number of eggs parasitized, number of emerged parasitoids, number of emerged parasitoids per parasitized egg, and percentage of emerged parasitoids that were female were the dependent variables. When significant (P < 0.05) effects were detected, the relationship between number of host eggs and the dependent variable was examined using a regression model The type of functional response was determined by performing a logistic regression of the proportion of parasitized eggs as related to their initial density. The type I functional response was estimated by the following linear equation: (1) Ne 0 where Ne = number of eggs parasitized, N0 the intercept and slope of the prediction line, respectively. The following curvilinear equation w as used to estimate the type II functional response: (2) Ne = aTN0/(1 + aThN0)
166 where Ne = number of eggs parasitized, N0 = number of host eggs, a = instantaneous search rate (area covered by a searching parasitoid in a given amount of time), T = total time of host exposure to the parasitoid, and Th = handling time. In the type III functional response, the instantaneous se arch rate (a) was a function of host density (N0) (Hassell et al. 1977) in the following relationship: (3) a = (d + b N0)/(1 = c N0) By substituting the value of a in E quation 3 to E quation 2, the following type III functional response equation was dev eloped: (4) Ne = d N0T = b N02T/(1 + c N0 + d N0Th = b N02Th) where parameters were described as above. Results The percentage of C. cactorum eggsticks parasitized by T. fuentesi varied from 45 to 90 but was not significantly influenced by egg density (from10 to 100 eggs per eggstick ) (F = 4.42; df = 1, 4, p = 0.10) ( Table 81). Although each female parasitoid was placed in close proximity to host eggs and enclosed in a relatively small arena (Petri dish, 3.5 x 1 cm), only 9% of the 6,000 total eggs ev aluated in this study were parasitized. Host egg densities had a significant effect on the number of eggs parasitized (F = 5.23; df = 5, 109, p = 0.0002) and the number of emerged parasitoids/eggstick (F = 2.85; df = 5, 109, p = 0.02) (Table 81). Host e gg densities also affected the number of emerged parasitoids/parasitized egg (F = 2.62; df = 5, 69, p = 0.03) but not the number of female parasitoids produced (F = 1.13; df = 5, 57, p = 0.36) (Table 81). The number of parasitized eggs as a function of e gg density appeared to display a sigmoid curve; however, statistical analysis demonstrated that the data fit a linear equation ( Ne = 1.37 + 0.0636N0) (Figure 8 2). Although the number of
167 parasitized eggs/eggstick and emerged parasitoids/eggstick (Figure 83) increased significantly with increasing host egg densities, these linear relationships explained very little (R2 = 0.12 and 0.05, respectively) (Table 82) of the variation in the data. Discussion Results from functional and numerical response studies are considered key information for evaluating the likelihood of success of an introduced biological control agent (Fernandez Arhex & Corley 2003; Lopes et al. 2008; Dannon et al. 2010). Functional response data provide information on the searching ability of a biological control agent at different host densities (Hassell 1978). A review of experimental studies on functional response of insect parasitoids from 1959 to 2001 (36 studies) identified only one example of a parasitoid [ Eretmocerus eremicus (Hyn enoptera: Aphelinidae)] exhibiting a type I response (Fernandez Arhex & Corley 2003). However, Trichogramma egg parasitoids were reported to display a type I response in more recent studies (Faria et al. 2000; Hoffmann et al. 2002; Mills & Lacan 2004). F or egg parasitoids intended as augmentative biological control agents, density dependent functional response, such as a type II or III is often recommended (Laumann et al. 2008). Parasitoids displaying a type II functional response are considered even more advantageous as biological control agents because they are more efficient at low pest densities than are those with type III strategies (Lopes et al. 2008). Ives et al. (1999) measured the behavioral response of Aphidius ervi (Haliday) (Hymenoptera: Braconidae) to its host, the pea aphid Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae), on two similar host plants containing the same number of aphids. They found that the parasitoid attack ed a higher number of hosts on one plant The variability in the number of attacks reduced the foraging efficiency of the
168 parasitoids, transforming a type II into a type I functional response (Ives et al. 1999; Fernandez Arhex & Corley 2003). In our study, T fuentesi did not show a significant varia tion in the attack rate of eggsticks at different host egg densities (Table 81), therefore the foraging efficiency was not affected when female T. fuentesi were presented with different numbers of host eggs. However the percentage of the 6,000 total host eggs parasitized across the different densities was low (9%), despite the fact that the female parasitoids were released close to their host eggs (Table 8 1). Although the reasons for this low attack rate from T. fuentesi are unknown, later studies ( Chapter 9 ) have show n that C. cactorum may not be a preferred host of T. fuentesi In addition, low levels of parasitism (< 0.2%) by T r i chogramma spp. on C. cactorum eggs were recorded in North Florida natural areas (Paraiso et al. 2011; Chapter 6). A number of experimental studies have focused on host finding and parasitism behaviors of Trichogramma spp. on major lepidopteran crop pests (Munyaneza & Obrycki 1997; Sithanantham et al. 2001; Hommay et al. 2002). These studies showed that efficiency of host searching was increased by the parasitoids ability to use infochemicals from its host and host plants. Conversely, little is known about parasitism mechanisms for egg parasitoids on lepidopteran pests foun d in natural areas. In nature, distribution of C. cactorum eggsticks on Opuntia plants is highly aggregated (Monro 1967). Field surveys conducted in North Florida showed that Trichogramma spp. preferentially attacked small sized C. cactorum eggsticks located on the areole/glochid structure of cactus pads (Paraiso et al. 2011; Chapter 6). T richogramma fuentesi displayed a type I functional response (Figure 81) to increasing numbers of C. cactorum eggs, suggesting that the attack rate increased
169 linearly. Although the relationship between the number of host eggs and the attack rate was described by a linear regression curve (Figure 81), statistical analysis revealed that the relationship was weak. The low R2 indicated that other factors were responsible for the main variation (Table 82). Such a functional response does not cause density dependent mortality for high host egg densities since the attack rate remained fairly constant. In our experiments, the search rate (i.e. the average encounters per hos t per unit searching time) for T. fuentesi among the different egg densities was low (0.063). In comparison, T. minutum Riley displayed a type I functional response to its preferred host, Ephestia kuehniella Zeller, and a search rate ranging from 0.37 to 1.75 (Mills & Lacan 2004). A positive numerical response was considered a useful attribute of an efficient biological control agent (Huffaker 1974). In this study, the number of parasitoid offspring produced per host egg generally increased with host de nsity (Table 81). A type I functional response, suggesting a lack of density dependence, was potentially compensated by the density dependent aspect of the numerical response by T. fuentesi T richogramma fuentesi had the ability to change its ovipositional behavior depending on host egg density. A population of T. fuentesi attacking C. cactorum eggsticks would have its highest growth rate at high host numbers. T richogramma fuentesi females reproduce by arrhenotokous parthenogenesis, in other words they have the ability to reproduce without mating and can manipulate the sex ratio of their offspring. Arrhenotokous females typically produce fertilized eggs that become diploid female parasitoids whereas unfertilized eggs become haploid male parasitoids (Flanders 1965). Therefore, sex ratio of the offspring is controlled by female
170 mating behavior (Nunney 1985; Luck et al. 1992) and the quality of the host (Charnov et al. 1981; Frank 198 6). The femalebiased sex ratio in Trichogramma contributes to reproductive success by allowing parasitoids to mate close to their host (Suzuki & Hiehata 1985). One of the advantages of local mating is that males will mate with their sisters, thereby reducing competition with brothers for mates (Hamilton 1967; Bulmer & Taylor 1980; Taylor 1981). Host size also influences sex ratio with a shift towards daughters occur in larger hosts due to increases in food resources (Charnov et al. 1981). In addition, the percentage of females produced in a patch of hosts is correlated with the fitness of the parent parasitoid (Hardy et al. 1998). Offspring production by T. fuentesi was characterized by a high percentage of females (Table 8 1). The mean percentage of female was not statistically different and did not increase at the various egg densities, suggesting that C. cactorum eggs were an acceptable host for T. fuentesi The low attack rate and type I functional response observed for T. fuentesi suggest s that this parasitoid would not be an appropriate c a ndidate for aug mentative biological control for C. cactorum particularly if the pest was present in high numbers Although the response by T. fuentesi on C. cactorum may not be density dependent, the high rat e of female to male offspring and the trend toward high numbers of parasitoids per parasitized egg suggest that T. fuentesi should be able to establish a stable population on C. cactorum eggs. However, the low level of parasitism suggests that T. fuentesi would not be an effective agent to use in inundative biological control programs against cactus moth.
171 Table 81. Parasitism of C. cactorum eggs at different densities by T. fuentesi (mean S.E. ). Mean # eggs/eggstick Proportion (%) eggsticks attacked (n = 20) # Parasitized eggs/eggstick # Parasitoids emerged/host egg % emerged females 10 9 ( 45) 1.05 0.35 2.90 1.07 77 10 20 14 ( 70) 2.95 0.65 9.20 2.08 68 10 40 5 ( 33) 2.46 1.60 9.80 6.27 59 19 60 14 ( 70) 8.45 2.05 23.65 7.07 79 5 80 16 ( 80) 6.25 1.21 13.70 2.80 86 3 100 18 ( 90) 6.40 1.02 16.25 3.84 80 6 *n = 15 Table 82. Linear regressions of the proportion of C. cactorum eggs parasitized by T. fuentesi with increasing densities of eggs Intercept R 2 Variable Parameter S.E. t Pr > t Parameter S.E. t Pr > t Eggs parasitized 1.37 0.99 1.38 0.17 0.0636 0.02 3.92 0.0002 0.12 Parasitoids emerged 5.49 3.35 1.64 0.10 0.1381 0.05 2.54 0.0126 0.05 Parasitoids emerged per parasitized host egg 3.75 0.44 8.58 0.0001 2.830 x 10 5 1.244 x 10 5 2.27 0.0259 0.13
172 Figure 81. Petri dishes set up to test functional response of individual Trichogramma fuentesi female parasitoids to various densities of Cactoblastis cactorum Trichogramma fuentesi females in small Petri dishes as shown here. Fig ure 82 Functional response of T richogramma fuentesi to different numbers of C actoblastis cactorum egg densities. The continuous line is the best fit to the data represented. Regression line assumes a Type I functional response. Details of regression line and parameters are given in Table 8 2.
173 Figure 83. The number of Trichogramma fuentesi emerged per parasitized eggs (mean S.E.) with different C actoblastis cactorum egg densities.
174 CHAPTER 9 TRICHOGRAMMA FUENTES I BEHAVIORIAL NOTES AND HOST SUITABILITY Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) is an important pest of Opuntia cac ti in North America (Zimmermann et al. 2001). Given its rapidly expanding geographical range, there is interest in exploring different control measures including the application of augmentative biological control. We conducted field surveys in North Flor ida to identify local natural enemies. These surveys led to the discovery of a naturally occurring parasitoid T richogramma fuentesi Torre (Hymenoptera: Trichogrammatidae), attacking C. cactorum eggs (Paraiso et al. 2011; Chapter 6). Trichogramma spp. ha ve been used in inundative programs against major lepidopteran pests (Li 1994). Not surprisingly, much is known about their biology and ecology (Flanders 1965; Takada et al. 2000; Doyon & Boivin 2006). However, biological and ecological characteristics vary considerably across species and between populations on different hosts. Egg characteristics, such as size, chorion configuration, and egg placement can affect host quality (Corrigan & Laing 1994; Greenberg et al. 1998). As such, the efficiency of Trichogramma spp. released as biological control agents (BCAs) was often related to its host egg characteristics (Ram & Irulandi 1989; Baorong et al. 1992). Host finding studies demonstrated that Trichogramma spp. females assessed host quality based on attributes such as egg shape, size, and external or internal chemical cues (Consoli et al. 1989), and that this information influenced the females allocation to her progeny (Schmidt & Smith 1985). The female examined a potential host by drumming her antennae on the host chorion for chemical cues (Schmidt & Smith 1986). Nonvolatile chemicals present on the surface of host eggs were perceived by receptors located on the antennae and tarsi of the females
175 (Godfray 1994). Female also walked on the host egg to assess surface area and diameter in order to estimate the host volume (Schmidt & Smith 1985, 1986). When an acceptable host egg was encountered, the female drilled into the host egg with her ovipositor which was covered in sensillae used to assess host suitability using internal chemical cues (Fisher 1971). The female laid one or more eggs depending on the host egg size (Hassan 1989). Studies showed that Trichogramma females attempted to parasitize any globular object between about 0.25 4.5 mm in diameter (Godfray 1994). Because Trichogramma spp.are generally polyphagous, c oncerns have been raised about possible detrimental impacts on nontarget host s during inundative releases ( Pinto et a l. 1986; Andow et al. 1995; Orr et al. 2000). Host specificity tests are a crucial component in determining the nontarget host assessment of the potential for entomophagous biological control agents (BCAs). Host specificity tests also help discern the p otential field success of BCAs used in classical introductions or inundative releases against pests in agrosystems or natural areas (Pluke & Leibee 2006; Yong & Hoffman 2006) R esults from host specificity tests help predict detrimental environmental impacts and address the safety of the proposed BCAs S election of nontarget species for host specificity testing must be carefully considered, as t he test results will give an indication on potential impacts on non target organisms (Blossey 1995; Greathead 1995; McEvoy 1996). Kuhlmann et al. (2006) published recommendations based on a comprehensive review of existing methods on selecting host species for host range testing of parasitoids being evaluated in classical biological control programs In the Kuhlm anns et al. (2006) method, an initial list is divided in a set of categories including ecological similarities and phylogenetic/taxonomic affinities.
176 Then, the list of non target species is reduced by eliminating those with different spatial, temporal, m orphological attributes, and species difficult to obtain (Kuhlmann et al. 2006). A similar process to Kuhlmanns et al. (2006), based on ecological, spatial, temporal and taxonomic similarities was developed and used for selection of host species for ent omophagous BCAs used in augmentative biological control. Although well implemented for herbivores used in weed biological control programs, the assessment of nontarget impacts for entomophagous BCAs is still in its infancy There is no a standardized pr ocedure for s pecificity testing for parasitoids (McEvoy 1996; Van Driesche & Hoddle 1997; Barratt et al. 2000). Laboratory host specificity studies have a higher possibility of giving either false positive or false negative outcomes (Kuhlmann et al. 2006) T wo types of experimental designs are used to assess host specificity (McEvoy 1996; Mansfield & Mills 200 4 ) : No choice test s assess the physiological host range which determines in the case of parasitoids the host species which are suitable to complet e development and c hoice test s which evaluate the realized field or ecological host range and indicate the preference of egg parasitoid s over acceptable hosts (McEvoy 1996; Mansfield & Mills 200 4 ). Against this background, the focus of this study was to: 1) Elucidate the developmental and reproductive biology of T. fuentesi on C. cactorum eggs; 2) Determine the key components of host finding behavior of the female parasitoids; 3) Carry out host specificity tests to evaluate the potential nontarget effects on the native cactus moth and butterfly eggs in the laboratory using a nochoice test.
177 Materials and Methods Trichogramma Rearing P rocedures Experiments were conducted at the USDA, Agricultural Research Service and FAMU, Center for Biological Control L aboratory Tallahassee, FL. Trichogramma fuentesi females used in this study were isolated from a rearing colony which originated from field collected parasitoids in North Florida. Parasitoid identity was confirmed by R. Stouthamer ( Department of Entomol ogy, University of California, Riverside, CA). Eggs of C cactorum from a mass rearing colony maintained on artificial diet were used as hosts for rearing and also as the source for experimental eggs To culture T. fuentesi h ost eggsticks containing par asitized eggs were glued onto note card strips (4 x 2 cm) with nontoxic Elmers glue (Elmers Products, Inc. Columbus, OH) The note card strips were then placed into standard plastic petri dishes (9 x 2 c m) lined with filter paper. A fresh raisin w as glued on a 1 x 1 cm note card in the center of the petri dish to provide a source of energy to emerging wasps. Newly emerged parasitoids were provided with food and maintained without access to hosts for 3 days in order to allow mating before conducting experiments. Based on previous studies, T fuentesi reared on C. cactoblastis eggs achieve their optimal oviposition activity 2 to 3 days after emergence (Chapter 7). Petri dishes were sealed with Parafilm (Pechiney Plastic Packaging, Menasha, WI) and arranged on plastic trays lined with moist wipes to increase relative humidity at 60 80% RH. The cultures were maintained in growth chamber at 2 8 1C, 16:8 L:D. Developmental and Reproductive Biology Development time of 20 randomly selected individual T. fuentesi females was studied using 1d old C. cactorum eggs. Twenty mated and fed females were randomly
178 selected and isolated from the r earing colony and placed in a petri dish (9 x 2 cm) lined with filter paper containing 1day old C. cactorum eggs. They were allowed to ovoposit over a 24h period. The rate of development for each stage was evaluated by observing changes in egg color at 24 h intervals. Previous studies showed that after parasitism, the host egg became tan when Trichogramma eggs hatched. Trichogramma larva fed internally and developed rapidly. During the last instar, dark melanin granules were deposited on the inner sur face of the egg chorion, causing the host egg to turn black. Larvae then transformed to an inactive pupal stage. After few days, the adult wasps emerged from the pupae and escape the host egg by chewing a circular hole in the egg shell (Ruberson & Kring 1993). Parasitoid emergence rate was determined by counting the number of emergence holes from black eggs. The sex ratio was determined by sex ing emerged dead adults under the microscope. Host Finding Behavior Study Host finding behavior of T. fuentesi females was studie d in a plastic p etri dish (30 x 10 mm) lined with filter paper under a stereoscopic microscope (KeyenceVH 5910) using online Windows Media to record all parasitoid behaviors. Behavioral events were scored using the event recording soft ware Observer XT version 8.0 (Noldus Information Technology, Wageningen, The Netherlands 2008). A mated and fed female parasitoid and a 1 day old C. cactorum eggstick with ten eggs were placed in the petri dish for a 10 h observation period (from 9 am to 7pm). The experiment was replicated five times with different randomly chosen females. Preliminary observations showed that T. fuentesi females displayed six behavioral events: walking, drumming, resting, grooming, drilling, and egg laying. The females did not host feed during the observations. The walking behavior started when the female left the surface of the host
179 egg for the filter paper and ended when she started drumming on the host egg Drumming occurr ed when the female examined the hos t by wal king on the egg surface while moving her antennae rapidly up and down (Schmidt & Smith 1989; Godfray 1994). The female occasionally became immobile which was described as resting and sometimes she would brush her antennae with her legs which corresponded to grooming. During drilling the female created a hole in the host by twisting her ovipositor from left to right. Egg laying behavior started when the female completely inserted her ovipositor into the host egg whi ch coincided with trembling movements o f the abdomen (corresponding to the passage of the egg through the ovipositor) (Suzuki et al. 1984). The g eneral sequence of oviposition behavior, total duration, mean duration, and timing of each behavior (the mean number of occurrences of a behavior per minute) was determined (Table 91) Host Specificity Tests Non t arget h ost s pecies s election The development of a nontarget host species list for T. fuentesi was based on recommendations developed by Kuhlmann et al. (2006) The initial list contained 22 species from 7 lepidopteran families (Table 92). The list was divided into six groups based on the following traits: taxonomic similarity phylogenetic similarity ecological affinities, endangered/threaten organism s, known weed natural enemies and other beneficial organisms, and organisms of economic value (Figure 9 1) No species were identified for group # 2, under phylogenetic similarity. At least one representative for each family in the remaining five groups was chosen The s pecies used in t he final list were selected based on ecological habitat, and temporal similarities of target and nontarget hosts A list of species tested can be found in Table 92. Several specimens of
180 nontarget speci es were purchased from a commercial butterfly rearing facility ( Old Oak Butterfly Farm Brooker, FL). D uring the development of the test species list entomologists, university researchers, federal scientists, and butterfly farm employees were consulted to ensure that all potential non target organisms w ere i nclud ed. No c hoice t est s Host acceptance and suitability was assessed by placing a single mated and honey fed female T. fuentesi with ten eggs of only one host test species into a plastic petri dish (30 x 10 mm ) lined with filter paper. Host eggs w ere arranged in the petri dish as single eggs and were less than 24h old when the parasitoid was introduced into the petri dish. Petri dishes were incubated (25 1 C, 16:8 L:D and 60% RH) until parasitoid emergence. The number of replications for each host test species depended on the availability of host eggs The number of parasitized eggs (as indicated by black coloration) per female, the number of emerged parasitoids per host egg and the percentage of emerged female parasitoids was recorded. Statis tical Analysis Data were analyzed for significant differences using analysis of variance using the SAS Statistical Software Version 9.2 (SAS Institute, Cary, NC). To compare t he percent parasitism between target and nontarget hosts, a nonparametric Tuk eys test was used. Estimates of central tendencies are recorded as mean standard error. Results Developmental and Reproductive Biology The egg, larval and p upal stages of T. fuentesi reared on eggs of C. cactorum lasted an average 1 0.07, 4 0.05 and 5 0.15 d, respectively. The sex ratio was female biased and the average percentage of females emerging from an eggstick was
181 74% 5 A dult female T. fuentesi provided with a source of carbohydrates lived an average of 11 days 0.79. Host Finding Behavior Female T. fuentes i include d six types of behavior s: walking, resting grooming, drumming, drilling, and egg laying. In general, the female wasp started the experiment by walking and searching for the host egg. Upon encounter ing the host, she drummed over the egg surface with her antennae, drilled through the chorion and oviposited into the host egg. Grooming was very infrequent (Table 91) Over a 60 min observation period, the female spent a significant portion of the time drilli ng (14.93 min) and laying eggs (19.66 min) (Table 9 1 ). Results showed that although time spent drilling (81.4 sec ) was almost twice as long as the time dedicated to egg laying (49.2 sec ), it was done less frequently with rates per minute ( the mean number of occurrences of a behavior per minute) of 0.18 and 0.40, respectively (Table 9 1 ). Host Suitability Experiment Trichogramma fuentesi attacked all host species tested from all experimental categories tested. The percent parasitism by T fuentesi on the non target host eggs ranged from 30 to 75% (Table 93). The highest level of parasitism was observed for Melitara prodenialis Walker (Lepidoptera: Pyralidae) (75% 2) and Dryas iulia (H bner) (Lepidoptera: Nymphalidae) (66% 5). The lowest level of parasitism was recorded for Cactoblastis cactorum (Berg) (11% 3.9). The statistical tests demonstrated that there was a difference in number of eggs parasitized among the various host species (F = 8.61, df = 6, 142, P < 0.0001). The level of parasitism f or the native cactus moth ( M. prodenialis ) was significantly higher than the nonnative cactus moth ( C. cactorum ) The number of progeny per host egg also was statistically different for the different
182 hosts (F = 11.32, df = 6, 142, P < 0.0001). T richogramma fuentesi exhibited gregarious tendency in most of the nontarget host species with more than two parasitoids emerging per host egg. In contrast, females presented a solitary tendency for Junonia coenia H bner ( Lepidoptera: Nymphalidae) and Van essa cardui L. ( Lepidoptera: Nymphalidae) eggs. Femalebiaised progeny were recovered from all experimental treatments (Table 93). Discussion The genus T richogramma includes more than 100 species (Voegele et al. 1988) which vary greatly in their searching behavior, and host preferences (Hassan 1989). The general sequence of ovipositional behavior of T fuentesi identified in this study differed from several other Trichogramma spp., such as T. platneri Nagarkatti, T pretiosum Riley, and T. brassicae Bezdenko (Blanch et al. 1996; Mills & Kuhlmann 2004) in that females did not host feed following oviposition. In general, the female wasp walked to a C. cactorum egg, drummed over the surface, drilled into the chorion and deposited an egg. Grooming and resting behaviors were observed very infrequently and host feeding was never recorded. The importance of host feeding among Trichogramma spp. is still unclear. S ome studies suggest that the behavior does not necessarily provide nutritional resources for egg production but serves as a way to restore lost water (Blanch et al. 1996; Nurindah & Gordh 1999; Mills & Kuhlmann 2004). Trichogramma fuentesi behavior was s imilar to T. platneri and T. pretiosum (Mills & Kuhlmann 2004) in that the species failed to oviposit in some host eggs after drumming but before drilling. Host rejection is based on both chemical and physical characteristics and generally happens before completion of host examination (De Jong & Pak 1984).
183 When a Trichogramma female finds preferred host eggs, it will usually stay on them until all or most of them are parasitized. Less preferable host eggs may be totally rejected or the female may lay fewer eggs (Hassan 1989). Mills & Kuhlmann (2004) demonstrated that T pretiosum and T. platneri spent a similar amount of time drumming and drilling on clusters of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs. In this study, although T. fuen tesi spent a longer time drilling into a C. cactorum host egg than T. pretiosum and T. platneri on their host, the females displayed a comparable examination time on a host egg. Following the general sequence of ovipositional behavior of T. fuentesi w e expected to observe a similar number of behavioral events per minute (rate per minute) for drilling and egg laying. When the host was considered suitable, female T. fuentesi drilled into the chorion and deposited an egg. However, the rate per minute for drilling (0.18) was significantly lower than the rate per minute for egg laying (0.40) (Table 91). These differences in rate might be explained by females often laying additional eggs into host eggs that had already been parasitized. In fact, several stu dies also have confirmed that Trichogramma spp. require greater drilling time during superparasitism (Mills & Kuhlmann 2004).The reason for this superparasitism or lack of host discrimination behavior is unclear. Our results suggested that, although C. c actorum was a suitable host it was less accepted than some of the other hosts tested. The native cactus moth, M. prodenialis was a well accepted and suitable host for T. fuentesi The number of eggs parasitized was much higher for the native cactus moth (Table 93). Trichogramma spp. are believed to be much more habitat specific than host specific (Smith 1996; Pinto 1998). The native cactus moth often cohabitates in the same plant host with C. cactorum and
184 both species have overlapping oviposition periods. Therefore, there is a high likelihood that the native host would be preferred during potential inundative releases of T. fuentesi In addition, T. fuentesi parasitized a si gnificantly higher number of butterfly eggs compared to C. cactorum eggs (Table 9 3). However, overall there was statistically no difference in the number of eggs laid per host egg between the nontarget hosts and C. cactorum (Table 93). The high rate o f parasitism for all nontarget species tested and the lowest rate of parasitism for the target species suggested that T. fuentesi should not be considered as a BCA for inundative releases against C. cactorum
185 Table 9 1. Time (seconds) and rate p er minute allocated by female T richogramma fuentesi over a 60 min observation period, for each ovipositional related behavior when associated with C actoblastis cactorum eggs. Measure Behavior Walking Grooming Resting Drumming Drilling Egg laying Total d uration (s) 641 101 309 663 896 1,180 Mean d uration (s) 12.5 6.3 12.87 10.5 81.4 49.2 Rate/ m inute 0.85 0.27 0.40 1.05 0.18 0.40
186 Figure 91. Methodology for selection of nontarget species based on Kuhlmann et al. 2006. Group 5: Natural enemies and other beneficial organisms Habitat Overlap Host with Morphological Affinities Test Species List External Peer Review Final Test Species List Group 1: Taxonomic a ffinities Group 2: Phylogenetic a ffinities Group 3: Ecological a ffinities Group 4: Endangered/ t hreaten o rganisms Group 6 : Organisms of economic values Filters Temporal
187 Table 9 2. List of Lepidoptera species developed for complete host specificity testing of Trichogramm a fuentesi Hosts are grouped by test considerations following Kuhlmann et al. (2006) Host Common Name Family Justifications Group 1 Taxonomic similarity Laetilia coccidivora (Comstock) Pyralidae Species taxonomically related in Florida Melitara prodenialis Walker Native cactus moth Pyralidae Species taxonomically related in Florida Ozamia lucidalis (Walker) Pyralidae Species taxonomically related in Florida Rumatha glaucatella (Hulst) Pyralidae Species taxonomically related in Florida Group 3 Ecological similarity Anaea floridalis Johnson & Comstock Florida L eaf W ing butterfly Nymphalidae Species occurring in the same habitat Danaus plexippus (L.) Monarch butterfly Danaidae Species occurring in the same habitat Melitara prodenialis Walker Native cactus moth Pyralidae Species occurring in the same habitat Papilio aristodemus ponceanus Schaus Schaus s wallowtail butterfly Papilionidae Species occurring in the same habitat Strymon acis bartrami Johnson & Comstock Bartrams H airstreak butterfly Lycaenidae Species occurring in the same habitat Group 4 Rare/Endangered species Anaea trogoglyta Florida L eaf W ing butterfly Nymphalidae Species rare or endangered in Florida Papilio aristodemus ponceanus Schaus Bartrams Hairstreak b utterfly Lycaenidae Species rare or endangered in Florida Strymon acis bartrami Johnson & Comstock Bartrams Hairstreak b utterfly Lycaenidae Species rare or endangered in Florida
188 Table 92. Continued Host Common Name Family Justifications Group 5 Important herbivores Sameodes albiguttalis (Warren) Pyralidae pyralid moth used as biological control of waterhyacinth Samea multiplicalis Guenee Pyralidae pyralid moth used against Salvinia and water lettuce plants Vogtia malloi Pastrana Pyralidae Alligatorweed stem borer, pyralid moth used against alligatorweed Group 6 Economic considerations Agraulis vanillae incarnata (Riley) Gulf Fritillary butterfly Nymphalidae Commonly reared and sold in Butterfly Farms in Florida Anartia jatrophae (L.) Peacock butterfly Nymphalidae Commonly reared and sold in Butterfly Farms in Florida Ascia monuste (L.) Great Southern butterfly Pieridae Commonly reared and sold in Butterfly Farms in Florida Danaus gilippus (Cramer) Queen butterfly Danaidae Commonly reared and sold in Butterfly Farms in Florida Danaus plexippus (L.) Monarch butterfly Danaidae Commonly reared and sold in Butterfly Farms in Florida Dryas iulia (H bner) Julia b utterfly Nymphalidae Commonly reared and sold in Butterfly Farms in Florida
189 Table 92. Continued Host Common Name Family Justifications Group 6 Economic considerations Heliconius charitonia (L.). Zebra Longwing butterfly Nymphalidae Commonly reared and sold in Butterfly Farms in Florida Junonia coenia (H bner) Buckeye b utterfly Nymphalidae Commonly reared and sold in Butterfly Farms in Florida Papilio cresphontes Cramer Giant Swallowtail butterfly Papilionidae Commonly reared and sold in Butterfly Farms in Florida Papilio glaucus (L.) Eastern Tiger Swall owtail butterfly Papilionidae Commonly reared and sold in Butterfly Farms in Florida Papilio polyxenes (F.) Black Swallowtail butterfly Papilionidae Commonly reared and sold in Butterfly Farms in Florida Phoebis sennae ebule (L.) Cloudless Sulfur butterfly Pieridae Commonly reared and sold in Butterfly Farms in Florida Vanessa cardui (L.) Painted L ady butterfly Nymphalidae Commonly reared and sold in Butterfly Farms in Florida
190 Table 93. Parasitism (Mean S.E. ) of potential host species by T richogramma fuentesi Host s pecies (10 eggs ) # r eplicat s % p arasitism # p arasitoids/ h ost e ggs % f emale s Cactoblastis cactorum 20 11 3.9 a 2.9 1.07 b 77 10 Group 1 Melitara prodenialis 13 75 2 f 3.8 0.4 c 60 7 Group 3 Danaus plexippus 9 58 3 d 2.3 0.5 b 68 4 Group 6 Dryas iulia 26 66 5 ef 3.1 0.4 bc 79 5 Junonia coenia 45 30 7 b 1.2 0.3 a 80 6 Papilio glaucus 5 26 5 b 3.8 1.1 c 100 Papilio polyxenes 5 45 2 c 3.0 1.1 b 69 4 Vanessa cardui 48 56 6 d 1.1 0.3 a 75 6 *Means with different letters are statistically significant according to Tukeys Least Mean Comparison test at P
191 CHAPTER 10 CONCLUSIONS Classical b iological control has been an accepted pest management tool worldwide for over a century but its implementation is not without risks. There is currently a divergence of opinions in the U.S. between decisionmakers and biological control practitioners on the appropriate way to assess the risk s asso ciated with the importation and release of entomophagous biological control agents ( BCAs ) The main objective of this dissertation was to improve the permitting process for the importation and release of entomophagous BCAs as implemented in 2007 in the U. S (Hunt et al. 2008). One suggestion made here for the development of an improved regulatory system was the implementation of efficient risk communication procedures during the Pest Risk Analysis (PRA). A survey was conducted on 500 biological control st akeholders among 5 professional affiliations (Federal, State, University, Private Sector, and Environmental groups) on risk communication during the permitting process for entomophagous BCAs. The objective of this survey was to characterize risk communica tion activities during the permitting process of entomophagous BCAs. In addition, the survey identified areas in the risk communication framework, as used in 2007, which could be improved. Results from the survey showed that the frequency of risk communi cation activities needs to be increased between decisionmakers and permit petitioners. Survey respondents wanted additional transfer of information pertaining to the PRA process of entomophagous BCAs. In addition, respondents believed that stakeholders should be involved during the decisionmaking process. The main difficulty of this study was to understand how to implement risk communication procedures within the permitting system. The proposed risk
192 communication procedures put forward in this disser tation are needed in order to improve the efficiency of the risk assessment process without hindering the permitting procedure. Consequently, a comparative study of regulatory systems for the importation and release of entomophagous BCAs in eight countries was done to identify risk communication procedures that could be easily implemented into the U.S permitting system. The International Standard for Phytosanitary Measures (ISPM) # 3 was used to compare important criteria of PRA. This study showed that t he selected countries have different approaches to risk assessment. The U.S. based their analysis on a guilty until proven innocent approach and the U.K. was considered highly risk adverse to the importation of BCAs. On the other hand, Australia based their assessment on a well detailed risk criteria and ranking system. In addition, unlike the U.S., the comparative analysis showed that most of the countries used a form of participatory/collaborative process which included expert consultation and public participation. Based on these findings, a collaborative PRA process was proposed to improve the permitting process for entomophagous BCAs in the U.S. The advantage of a collaborativebased risk analysis process is the increase of transparency during the decision making process. In addition, this type of approach will decrease epistemic uncertainties by using recommendations from experts. In 2007, a permit (PPQ 526) was required by USDA APHISPPQ for the importation, interstate movement, and environmental release of entomophagous BCAs. Information as described in the Regional Standard for Phytosanitary Measures # 12 was also required from the permit petitioner. In addition, recommendations from representatives from the North American Plant Protection Or ganization (NAPPO) were solicited. However, there
193 was not a formalized review committee such as the one used for the permitting process for phytophagous BCAs so that a published decision could be made on all of the petitioners information. To increase decision making transparency, one recommendation of this study was the development of an Environmental Assessment (EA) under NEPA (National Environmental Policy Act) which would publish all relevant information and require public comments. In addition, in t he modified permitting process, an additional step for external review from an expert committee was suggested. During the course of this dissertation, several major regulatory reforms have occurred. In 2009, a proposed rule solicited public comments about the regulations related to permitting conditions as established in 7 CFR section 330 Parts 200212 for entomophagous BCAs (CFR 2001). As of 2011, many of the regulatory changes proposed in this dissertation are currently in place. An EA is required for all permit petitioners, prior to environmental release of entomophagous BCAs (APHIS 2011). In addition, a sort of advisory committee is consulted by USDA APHISPPQ during the decisionmaking process (APHIS 2011). Another recommendation made in this dis sertation for the development of a more efficient permitting process for entomophagous BCAs was the implementation of harmonized sciencebased procedures for the evaluation of risks. Several risk assessment procedures have been published attempting to estimate potential environmental and economic deleterious impacts of proposed BCA s ( van Lenteren et al. 2003, van Driesche & Reardon 2004; Bigler et al. 2006). This dissertation used a real case study to assess environmental risks associated with the use of an entomophagous BCA. The cactus moth, Cactoblastis cactorum (Berg), is an invasive moth native to
194 South America attacking Opuntia cacti in the U.S. Field surveys conducted in Argentina identified an egg parasitoid from the family Trichogrammatidae attacking C. cactorum eggs. However, in 2008, because of the stringent requirements, lengthy permitting process, and time constraint inherent to this doctoral study, natural enemies already present in the U.S. were used for the evaluation. Consequently, field surveys were conducted in six locations in North Florida. Egg parasitoids from the family Trichogrammatidae, Trichogramma pretios um Riley, Trichogramma fuentesi Torre and an unidentified species in the T. pretiosum group were found attacking C. cactorum eggs. When implementing a classical biological control program to control an invasive pest, one major step consists of assessing possible nontarget impacts. In addition, evaluation of important biological parameters is required to assess whether the entomophagous BCA is a suitable agent. Using the same rationale, this dissertation used upto date methods for evaluating environmental risk of T. fuentesi against C. cactorum During risk evaluation, the best rearing conditions for the proposed BCA colony was determined to obtain accurate results. The presence, type of food source, and mating status were determined to have an influence on the fitness and percent of parasitism by T fuentesi on C. cactorum The age range for both the egg parasitoid and target host were assessed and the optimal ages for optimal percent parasitism by T. fuentesi were identified. Data from the functional response experiment of T. fuentesi provided information on the females searching ability and attack rate efficiency. The density independence of T. fuentesi s functional response was compensated by the density dependence of the wasps numerical respons e. This experiment showed that
195 data from the numerical response also needed to be included in the assessment of this proposed BCA. For the evaluation of possible nontarget impacts, s tep by step procedures for the development of host lists usually includ e species with similar biological, ecological, and taxonomic attributes to the target host In this study, the non target host species list used was developed using a modified version of the Kuhlmann et al. (2006) method. One important addition in the pr ocedure was the consultation of an expert group during the development of the test species list. The group of expert s was composed of entomologists, university researchers, federal scientists, and butterfly farm employees. This additional step assured th at all potential nontarget organisms were identified. However, the evaluation of host acceptance and suitability was limited by the availability of the hosts. In addition, the appropriate number of replications was difficult to determine. A consistent assessment of environmental impacts of a potential entomophagous BCA relied on the accurate selection of nontarget species for testing, the appropriate experimental design to run host specificity tests, and the correct interpretation of results. In 2007, the existing permitting process for the importation and release of entomophagous BCAs was considered inadequate by the biological control community. The ISPM # 3 describes responsibility and data requirements of parties involved prior to importation, ship ment and release of entomophagous BCAs. However, it does not provide guidance on rigorous methods that might be used during risk assessment. This dissertation showed that to improve the efficiency of the permitting process, efforts must be given to both the regulatory system and the risk assessment process. There is a need for the development of a harmonized guidance document with robust procedures
196 for the evaluation of potential detrimental environmental impacts from entomophagous BCAs.
197 APPEND IX A INTERVIEW INITIAL LETTER Florida Agricultural and Mechanical University Tallahassee, Florida 323073100 CENTER FOR BIOLOGICAL CONTROL T elephone: (850) 4127062 310 Perry Paige Build. (South) Fax: (850) 5612221 June 11th, 2007 Dr. XXXXXX UF Department of Entomology PO Box 110680, Gainesville, Fl 326110680 Dear Dr. XXXXX, Risk Communication in Biological Control The Center for Biological Control at Florida A& M University is conducting research on aspects of risk analysis for entomophagous biological control agents. To this end we have a graduate student undertaking part of the research. One component of this work is focused on aspects of Risk Communication dur ing the permitting process for entomophagous biological control agents with a view to develop an improved process. This letter is to introduce the student, Ms. Oulimathe Paraiso, who is registered in the joint Cooperative Ph.D. Program between Florida A& M University and University of Florida, with Dr. Stephanie Bloem and myself as her major supervisors. Other members of her committee are: Drs. James Cuda, Norm Leppla, Robert McGovern and Marcia Owens. As part of the research she will undertake a broad s urvey to better understand and quantify current practices and perceptions of both decisionmakers and stakeholders. As the first step in this process of identifying the key issues, she plans to get the perspectives from a limited number of key decision mak ers and stakeholders (20 in totals) in order to help shape the survey instrument that will be distributed to broad spectrum of representatives from both groups. Because of your extensive background and knowledge of the issues, she plans to request your ass istance during this first stage. We hope you will be able to participate in this research. With thanks. Yours sincerely, Moses T.K. Kairo Director/Associate Professor
198 APPENDIX B INTERVIEW PRE NOTICE LETTER Florida Agricultural and Mechanical University Tallahassee, Florida 323073100 CENTER FOR BIOLOGICAL CONTROL T elephone: (850) 4127062 310 Perry Paige Build. (South) Fax: (850) 5612221 June 18th, 2007 Dr. XXXXXXXXX UF Department of Entomology PO Box 110680, Gainesville, Fl 326110680 Risk Communication in Biological Control I am writing to follow up on the letter sent by Dr. Moses Kairo on June 11th 2007 and would like to request your assistance during this initial phase. I am specifically interested in un derstanding the different mental models associated with Risk Communication procedures employed during the importation of entomophagous biological control agents (BCAs). Understanding the different mental models used by decisionmakers and stakeholders will contribute to the development of an improved Risk Communication framework. R isk analysis (RA) the assessment of the likelihood of occurrence of an adverse outcome and the magnitude of the consequences of th is outcome is composed of three major tasks: Ri sk Assessment, Risk Management, and Risk Communication. Guidelines on importation of BCAs have been developed at the international level (International Standard for Phytosanitary Measure # 3) and at the regional level (Regional Standard for Phytosanitary M easure # 12). The USDA Animal and Plant Health and Inspection Service ( USDA APHIS) in the context of safeguarding plant life and health have developed their own guidelines to conduct PRA. Implicit in these guidelines is the need for conducting a risk analy sis prior to importation of BCAs. Thus an important component of the importation process is risk communication. Mental models are representations of real or imaginary situations They are used to anticipate an outcome. The main source of mental
199 models is perception. Mental models shape decisions because they affect decisionmakers perceptions and beliefs. The decisionmaking process can be described as an interactive process between decisionmakers and stakeholders. Risk C ommunication inten ds to supply decision makers and stakeholders with the information they need to make informed independent judgments about risks. Therefore, i t becomes an important tool to prevent unwanted outcomes. Recommendations for better Risk C ommunication are made in the ISPM #3 H owever, while R isk C ommunication is an essential component of the decisionmaking process it is often taken for granted. The purpose of the current research project is to assess the mental models applied by decisionmakers and stakeholders in the context of the Risk C ommunication procedures that take place during the entomophagous BCA permitting process. The approach used is based on methods developed by Morgan et al., 2002. This involves a series of five sequential steps as follow s: 1. C reate an expert model based on scientific data to determine the nature and magnitude of the problem 2. C onduct interviews to elicit peoples beliefs 3. C reate a confirmatory questionnaire to estimate the population prevalence of these beliefs 4. U se the results to determine which inc orrect beliefs must be corrected. 5. E valuate communication by testing the improved communication framework. I am now at the second step and you have been identified as part of a carefully selected group of 20 people. All 20 persons are knowledgeable about t he importation of BCAs. I have developed a short questionnaire and would also like to interview you after you have had a chance to respond. This step will essentially assist me to identify the critical issues in risk communication. Based on the information gathered during this step, I will develop a comprehensive questionnaire which will be sent to a broad range of stakeholders (1000 1500 stakeholders). The short questionnaire is being sent to you electronically and can be completed directly on the web. I will also mail to you a hardcopy if you prefer this medium. Please be assured that your answers to this survey will be completely confidential and they will only be released as summaries in which no individuals answers will be identifiable. Should you have any concerns with or difficulties in responding to this survey please contact me at (850) 4127062 or at firstname.lastname@example.org Sincerely, Oulimathe Paraiso Ph.D. Student
200 APPENDIX C INTERVIEW BOOKLET
209 APPENDIX D EXAMPLES OF MENTAL MODELS FOR THE PERMITTING PROCESS FOR ENTOMOPHAGOUS BIOLOGICAL CONTROL AGENTS IN THE U.S.
210 APPENDIX E REMINDER NOTICE Dear Dr. XXXXXX You were recently sent a survey concerning Risk Communication during the Permitting Process for Entomophagous Biological Control Agents. If you have already completed the survey, we would like to thank you for your time and important contribution. If you h avent had a chance to complete the survey yet, please take a moment to answer the questions at this url address: http://is nri.com/take?i=116471&h=TiOSnMO6tP6KD6p7VlFXWg Your exper tise will help us document and characterize Risk Communication activities during the permitting process. If you have any questions concerning this survey, you may contact me at (850) XXX XXX or XXXXX@ufl.edu Sincerely, Oulimathe Paraiso Ph.D. Student Center for Biological Control Florida A&M University
211 APPENDIX F QUESTIONNAIRE
218 APPENDIX G INSTITUTIONAL REVIEW BOARD APPROVAL LETTER
219 LIST OF REFERENCES ABATE, T., VAN HUIS, A., AND AMPOFO, J. K. O. 2000. Pest management strategies in traditional agriculture: An African perspective. Ann. Rev. Entomol. 45: 631 659. A MALIN, D M P ENA, J. E ., AND D UNCUN, R. 2005. Effects of host age, female parasitoid age, and host plant on parasitism of Cer atogramma etiennei (Hymenoptera: Trichogrammatidae). Florida Entomol. 88: 77 81. ANDERSEN, M. C., ADAMS, H., HOPE, B., AND POWELL, M. 2004. Risk assessment for invasive species. Risk Anal. 24: 787 793. ANDOW, D. A., KLACAN, G. C., BACH, D., AND LEAHY, T. C. 1995. Limitations of Trichogramma nubilale (Hymenoptera: Trichogrammatidae) as an inundative biological control of Ostrinia nubilalis (Lepidoptera: Crambidae). Environ. Entomol. 24: 1352 1357. APHIS (ANIMAL PLANT HEALTH AND INSPECTION SERVICE). 1 996. Options for changes in biological control regulations and guidelines in the United States: A Strawman for comment. National Biological Control Institute, Riverdale, MD. APHIS (ANIMAL PLANT HEALTH AND INSPECTION SERVICE). 2006. Plant protection and quarantine permitting review highlights, DA 200604. [Online] http://ipm.ifas.ufl.edu/pdf/OrganismsPermittingReview.pdf Last accessed 15/12/2010. APHIS (ANIMAL AND PLANT HEALTH INSPECTION SERVICE). 2007. United States Department of Agriculture, Animal Plant Health Inspection Service, Imports and Exports [Online] www.aphis.usda.gov Last accessed 09/01/2007. APHIS (ANIMAL AND PLANT HEALTH INSPECTION SERVICE). 2009. APHIS Web Survey [Online] www.surveymonkey.com/s.aspx?sm=WuAlyS1l8brM_2fUYhlRLISw_3d_3d. Last accessed 11/19/2009. APHIS (ANIMAL AND PLANT HEAL TH INSPECTION SERVICE). 2011. United States Department of Agriculture, Animal Plant Health Inspection Service, Imports and Exports [Online] http://www.aphis.usda.gov/permits/ Last accessed 03/18/2011. AQ IS (AUSTRALIA QUARANTINE INSPECTION SERVICE). 1997. Importing to Australia [Online] www.daff.gov.au/aqis/import. Last accessed 01/31/11. ARROW, K. J., AND FISHER, A. C. 1974. Environmental preservation, uncertainty, and irreversibility. Q. J. Econ. 88: 312 319.
220 ASHLEY, T. P., ALLEN, J. C., AND GONZALES, D. 1974. Successful parasitism of Heliothis zea and Trichoplusia ni eggs by Trichogramma. Environ. Entomol. 3: 319 322. AUSTIN D. F., BINNINGER, D. M., AND PINKAVA, D. J. 1998. Uniqueness of the endangered Florida semaphore cactus ( Opuntia corallicola ). Sida 18: 527 534. BAI, B., AND SMITH, S. M. 1993. Effect of host availability on reproduction and survival of the parasitoid wasp Trichogramma mi nutum Ecol. Entomol. 18: 279 286. BALCIUNAS, J. K. 1990. Australian insects to control Melaleuca. Aquatics 12: 15 19. BAORONG, B., LUCK, R. F., FORSTER, L., STEPHENS, B., AND JANSSEN, J. A. M. 1992. The effect of host size on quality attributes of the egg parasitoid, Trichogramma pretiosum Entomol. Exp. Appl. 64: 37 48. BARRATT, B. I. P., EVANS, A. A., FERGUSON, C M., BARKER, G. M., MCNEILL, M. R., AND PHILLIPS, C. B. 1997. Laboratory nontarget host range of the introduced parasitoids Microctonus aethiopoides and M. hyperodae (Hymenoptera: Braconidae) compared with field parasitism in New Zealand. Environ. Entomol. 26: 694 702. BARRATT, B. I. P., EVANS, A. A., FERGUSON, C. M., MCNEILL, M. R. PROFFITT, J. R., AND BARKER, G. M. 1998. Curculionoidea (Insecta: Coleoptera) of agricultural grassland and Lucerne as potential nontarget hosts of the parasitoids Microctonus aethiopodes Loan and Microctonus hyperodae Loan (Hymenoptera: Braconidae). New Zealand J. Zool. 25: 47 63. BARRATT, B., GOLDSON, S. L., FERGUSON, C. M., PHILIPS, C. B., AND HANNAH, D. J. 2000. Predicting the risk from biological control agent introductions: A New Zealand approach, pp. 59 76 In P.A. Follett and J.J. Duan [eds.], Nontarget Effects of Biological Control. Kluwer Academi c Publishers, Norwell, MA. BARRATT, B. I. P., AND MOEED, A. 2005. Environmental safety of biological control: Policy and practice in New Zealand. Biol. Control 35: 247 252. BENNETT, F. D., AND HABECK, D. H. 1995. Cactoblastis cactorum : A successful weed control agent in the Caribbean, now a pest in Florida? pp. 21 26 In E.S. Delfosse and R.R. Scott [eds.], Proc. VIIIth Inter. Symp. Biol. Control Weeds 2 7 February 1992, Canterbury, NZ. BENSON, L. 1982 The Cacti of the United Stat es and Canada, Stanford University Press, Stanford, CA. BERNAL, J. S., BELLOWS, JR., AND GONZALES, T. S. 1994. Functional response of Diaeretiella rapae (McIntosh) (Hymenoptera: Aphididae) to Diuraphis noxia (Morawiko) (Homoptera: Aphididae) hosts. J. A ppl. Entomol. 118: 300 309.
221 BERNAYS, E. A. 2000 Neural limitations in phytophagous insects: Implication for diet breadth and evolution of host affiliation. Ann. Rev. Entomol. 46: 703 727. BERRYMAN, A. A. 1999. The theoretical foundations of biological control, pp. 3 21 In B.A Hawkins and H.V. Cornell [eds.], Theoretical Approaches to Biological Control. Cambridge University Press, Cambridge. BERTI, J., AND MARCANO, R. 1993. Effect of different food of substances on the reproduction and lifespan of the female of Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). Bol. Entomol. Venezolana 8: 105 110. BIGLER, F., B ABENDREIER D. AND K UHLMANN, U. 2006. Environmental Impact of Invertebrates in Biological Control of Arthropods: Methods and Risk Assessment C AB Int Wallingford, UK BIGLER, F., BALE, J. S., COCK, M. J. W., DREYER, H., GREATREX, R., KUHLMANN, U., LOOMANS, A. J. M., AND LENTEREN, J. C. 2005. Guidelines on Information Requirements for Import and Release of Invertebrate Biological Control Agents in European Countries. CAB Reviews Perspectives in Agriculture, Veterinary Sciences, Nutrition and Natural Resources 1.10. BLAKENEY, R .L. 2002. Providing Relief to Families after a Mass Fatality: Roles of the Medical Examiner s Office and the Family Assistance Center. O.V.C. Bulletin, November 2002. U.S. Department of Justice, Office of Justice Programs, Office for Victims of Crime, Washington, D.C. BLANCH, S., CASAS, J., BIGLER, F., AND JANSSEN VAN BERGEIJK, K. E. 1996. A n individual based model of Trichogramma foraging behavior: Parameter estimation for single females. J. Appl. Ecol. 33: 425 434. BLOEM, K., BLOEM, S., CARPENTER, J., HIGHT, S., FLOYD, J., AND ZIMMERMANN, H. 2007. Dont let cacto blast US Development of a bi national plan to stop the spread of the cactus moth Cactoblastis cactorum in North America, pp. 337 344 In M.J.B. Vreysen, A.S. Robinson, and J. Hendrichs [eds.], Area Wide Control of Insects Pests: From Research to Field Implementation. Springer, Dordrecht, The Netherlands. BLOEM, S., BLOEM, K., AND KNIGHT, A. L. 1998. Oviposition by sterile codling moths, Cydia pomonella (Lepidoptera: Tortricidae), and control of wild populations with combined releases of sterile moths and egg parasitoids. J. Ent omol. Soc. British Columbia 95: 99 109. BLOEM, S., HIGHT, S. D., CARPENTER, J. E., AND BLOEM, K. A. 2005. Development of the most effective trap to monitor the geographical expansion of the cactus moth Cactoblastis cactorum (Lepidoptera: Pyralidae). Flor ida Entomol. 88: 300 306.
222 BLOSSEY, B., SCHROEDER, D., H IGHT, S. D., AND M ALECKI, R. 1994. Host specificity and environmental impact of two leaf beetles ( Galerucella calmariensis and G. pusilla ) for biological control of purple loosestrife ( Lythrum salicaria ) Weed Sci. 42: 128 133. BLOSSEY, B. 1995. A comparison of various approaches for evaluating potential biological control agents using insects on Lythrum salicaria. Biol. Control 5: 113 122. BOIVIN, G., KLLIKER OTT, U. M., BALE, J. S., AND BIGLER, F. 2006. Assessing the establishment potential of inundative biological control agents, pp. 98 113 In F. Bigler, D. Babendreier, and U. Kuhlmann [eds.], Environmental Impact of Invertebrates for Biological Control of Arthropods: Methods and Risk Assessm ent. CAB International, Wallingford, UK. BOSTROM, A., FISCHNOFF, B., AND MORGAN, M. G. 1992. Characterizing mental models of hazardous processes: A methodology and an application to radon. J. Soc. Issues 48: 85 100. BOYD, E. A., AND HODDLE, M. S. 200 7. Host specificity testing of Gonatocerus spp. egg parasitoids used in a classical biological control program against Homalodisca vitripennis : A retrospective analysis for nontarget impacts in southern California. Biol. Control 43: 56 70. BRYAN, M. D., DYSART, R. J., AND BURGER, T. L. 1993. Releases of I ntroduced P arasites of the A lfalfa W eevil in the United States, 1957 88. USDA APHIS Misc. Publ. No. 1504. Washington, DC. BULMER, M. G., AND TAYLOR, P. D. 1980. Dispersal and the sex ratio. Nature 284: 448 449. BURGMAN, M. A. 2005. Kinds of uncertainty, pp. 26 41 In M.A. Burgman [ed.], Risks and Decisions for Conservation and Environmental Management. University Press, Cambridge. CALTAGIRONE L. E., A ND DOUTT R. L 1989. The history of the Vedalia beetle importation to California and its impact on the development of biological control. Ann. Rev. Entomol. 34: 1 16. CARPENTER, J. E., BLOEM, S., AND MARE C F. 2005. Inherited sterility in insects p p 39 68 In V.A. Dyck, J. Hendricks and A.S. Robinson [e ds. ] Sterile Insect Technique, Principles and Practice in Area Wide Integrated Pest Management, IAEA. Springer Dordrecht. The Netherlands. CFIA (CANADIAN FOOD AND INSPECTION AGENCY). 2006. Applic ation, Procedures and Use of a Permit to Import. [Online] http://www.inspection.gc.ca/english/plaveg/protect/dir/d9704e.shtml Last accessed 09/01/2007.
223 CGIAR (CONSULTA TIVE GROUP ON INTERNATIONAL AGRICULTURAL RESEARCH). 2008. [Online] www.cgiar.org Last accessed 06/24/2008. C HARNOV, E L L OS DEN HARTOGH, R L J ONES, W T AND VAN DEN ASSEM, J. 1981. Sex ratio evolution in a variable environment. Nature 299: 27 33. CHARUDATTAN, R. 2001. Biological control of weeds by means of plant pathogens: Significance for integrated weed management in modern agroecology. Biol. Control 46: 229260. CHESS, C., SALOMONE, K. L., AND HANCE, B. J. 1995. Improving risk communication in government: Research priorities. Risk Anal. 15: 127 135. CIA (CENTRAL INTELLIGENCE AGENCY). 2011. The World Factbook. [Online] https://www.cia.gov/library/publications/theworld factbook/ Last accessed 04/01/2011. CLAUSEN, C. P. 1940. Entomophagous Insects. McGraw Hill Book Co., New York, NY. CFR ( CODE OF FEDERAL REGISTRY ) 2001. Plant Protection Act, Commerce and Trade, Exports and Imports [Online] http://www.aphis.usda.gov/brs/pdf/PlantProtAct2000.pdf Last accessed 04/01/2011. CONSOLI, F. L., KITAJIM A, E. W., AND PARRA, J. R. P. 1989 Ultrastructure of the natural and factitious host eggs of Trichogramma gallao Zucchi and Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). Int. J. Insect Morphol. 28: 211 231. CORNELIUS, M. L., GRACE, J. K., AND YATES, J. R. 1996. Acceptability of different sugars and oils to three tropical ant species (Hymen. Formicidae). Anz. Schadlingsdk. Pfl. 69: 41 43. CORRIGAN, J. E., AND LAING, J. E. 1994. Effects of the rearing host species and the host species attacked on performance by Trichogramma minutum Riley. Environ. Entomol. 23: 755 760 COSAVE (EL COMITE DE SANIDAD VEGETAL DEL CONO SUR). 1996. Control Biologico Actas [Online] www.cosave.org/documentos oficiales.php?ver=actas Last accessed 09/01/2007. DAFF (DEPARTMENT OF AGRICULTURE FISHERIES, AND FORESTRY). 2007. AQIS Forms. [Online] h ttp://www.daff.gov.au/aqis/import/application/forms/biological materials Last accessed 09/01/2007.
224 DAHLSTEN, D. L., AND MILLS N. J 1999. Biological control of forest insects, pp. 761 788 In T.S. Bellows and T.W. Fisher [eds.] Handbook of Biological Control Academic Press, San Diego. DANIELS, S. E., AND WALKER, G. B. 2001. Collaboration as a deliberative process, pp. 55 76 In S.E. Daniels and G.B. Walker [eds.], Working Through Environmental Conflict: The Collaborative Learning Approach. Praeger, Westport, Connecticut. DANNON, E. A., TAM, M., VAN HUIS, A., AND DICKE, M. 2010. Functional response and life history parameters of Apalantes taragame, a larval parasitoid of Maruca vitrata BioControl 55: 363 378. DAVIS, J. B. 2007. Inference on th e means and variances from two independent populations, pp. 281 284 In J.B. Davis [ed.], Statistics Using SAS Enterprise Guide. SAS Institute Inc., Cary, NC. DAY, W. H. 1981. Biological control of the alfalfa weevil in the Northeastern United States. Chap. 25 In G.C. Papavizas [ed.], Biological Control in Crop Protection. Allanheld, Osmun & Co., Montclair, NJ. DEFRA (DEPARTMENT FOR ENVIRONMENT, FOOD AND RURA L AFFAIRS). 2000. Non Native Species. [Online] http://ww2.defra.gov.uk/ Last accessed 09/01/2007. DEBACH, P. 1973. Biological Control of Insect Pest and Weeds. Chapman and Hall, London, UK. DEH (DEPARTMENT OF ENVIRONMENT AND HERITAGE). 2007. Permits and Application Forms. [Online] http://www.environment.gov.au/epbc/permits/index.html Last accessed 09/01/2007. DE JONG E. J., AND PAK, G. A. 1984 Factors determining differential host egg recognition of two hosts species by different Trichogramma spp. Meded. Fac. Landbouww. Rijksuniv. Gent. 49: 815 825. D EKAY M., SMALL, M J., FISCHBECK, P S., FARROW, R. S., CULLEN, A KADANE, J. B. L AVE, L B., MORGAN, M. G ., AND TAKEMURA, K 2002. Risk based decision analysis in support of precautionary policies. Pol. Reg. 221 247. DELFOSSE, E. S. 2005. Risk and ethics in biological control. Biol. Control 35: 319 329. DE NARDO, E. A. B., AND HOPPER, R. K. 2004. Using the literature to evaluate parasitoid host ranges: A case study of Macrocentrus grandii (Hymenoptera: Braconidae) introduced into North America to control Ostrinia nubilalis (Lepidoptera: Crambidae). Biol Control 31: 280 295.
225 DJC (DEPARTMENT OF JUSTICE CANADA). 2005. The Plant Protection Act 1990, c.22. [Online] http://laws.justice.gc.ca/eng/. Last accessed 09/01/2007. DICKEL, T.S. 199 1. Cactoblastis cactorum in Florida (Lepidoptera: Pyralidae: Phycitinae). Trop. Lepidoptera 2: 117 118. D ILLMAN, D A 2000 Mail and I nternet S urveys, t he T ailored D esign Method. John Wiley & Sons, Inc ., New York, NY DIXIT, A. K., AND PINDYCK, R. S. 1994. Investment u nder Uncertainty Princeton University Press, Princeton, NJ. DODD, A. P. 1940. The Biological Campaign Against Prickly Pear. Commonwealth Prickly Pear Board, Brisbane, Australia. DONER, L. W. 1977. The sugars of honey a review. J. Sci. Food Agr. 28: 443 456. DOYON, J., AND BOIVIN, G. 2006. Impact of the timing of male emergence on mating capacity of males in Trichogramma evanescens Westwood. Biocontrol 51:703 713. DUAN, J. J., AND MESSING, R. H. 199 6. Response of two opiine fruit fly parasitoids (Hymenoptera: Braconidae) to the lantana gall fly (Diptera: Tephritidae). Env. Entomol. 25: 1428 1437. DUAN, J. J., AND MESSING, R. H. 2000. Evaluating non target effects of classical biological control: Fruit fly parasitoids in Hawaii as a case study, pp. 95 109 In P.A Follett and J.J. Duan [eds.], Non target Effects of Biological Control Kluwer Academic Publishers, Norwell, MA. DUAN, J. J, MUKHTA, A., KAILASH, J., AND MESSING, R. H 1997 Evaluation of the impact of the fruit fly parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae) on a nontarget tephritid, Eutreta xanthochaeta (Diptera: Tephritidae). Biol. Control 8: 58 64. EBBELS, D. L. 2003. Early history of plant health control measures, pp. 929 In D.L. Ebbels [ed.], Principles of Plant Health and Quarantine. CABI Publishing Wallingford Oxon, UK. EILENBERG, J., HAJEK, A., AND LOMER, C. 2001. Suggestions for unifying the terminology in biological control. Biocontrol 46: 3874 00. EPPO (EUROPEAN AND MEDITERRANEAN PLANT PROTECTION ORGANIZATION). 1999. First Importation of Exotic Biological Control Agents for Research under Contained Conditions (PM6/1). [Online] http://archives.eppo.org/EPPOStandards/biocontrol.htm Last accessed 03/24/11.
226 EPPO (EUROPEAN AND MEDITERRANEAN PLANT PROTECTION ORGANIZATION). 2001a. Import and Release of NonIndigenous Biological Control Agents (PM6/2). [Online] http://archives.eppo.org/EPPOStandards/biocontrol.htm Last accessed 03/24/11. EPPO (EUROPEAN AND MEDITERRANEAN PLANT PROTECTION ORGANIZATION). 2001b. Pest Risk Analysis (PM5). [Online] http://archives.eppo.org/EPPOStandards/pra.htm Last accessed 03/24/11. EPPO (EUROPEAN AND MEDITERRANEAN PLANT PROTECTION ORGANIZATION). 2006. EPPO Activities on Plant Quarantine. [Online] http://www.eppo.org/QUARANTINE/quarantine.htm Last accessed 03/24/11. ERMA (ENVIRONMENTAL RISK MANAGEMENT AUTHORITY). 1997a. Import New Organisms in a Containment Facility. [Online] http://www.ermanz.govt.nz/new organisms/findapplicationform/all applications/Pages/default.aspx Last accessed 03/24/11 ERMA (ENVIRONMENTAL RI SK MANAGEMENT AUTHORITY). 1997b. Releasing New Organisms into the New Zealand Environment. [Online] http://www.ermanz.govt.nz/new organisms/f ind applicationform/all applications/Pages/default.aspx Last accessed 03/24/11. ERMA (ENVIRONMENTAL RISK MANAGEMENT AUTHORITY). 2006. New Organisms [Online] http://www.ermanz.govt.nz/. Last accessed 03/24/11. E RTLE L.R. 1993. What quarantine does and what the collector needs to know, pp. 5365 In R. G Van Driesche and T.S Bellows [ eds. ], Steps in Classical Arthropod Biological Control. Entomological Society of America, Lanham Maryland. FAO (FOOD AND AGRICUL TURE ORGANIZATION). 2007. Plant Protection Profiles from Asia Pacific Countries. Food and Agricultural Organization of the United Nations, Rome. [Online ] www.fao.org/docrep/012/i0972e/i0972e00.htm Last accessed 03/24/11. FAIRCHILD, D. 1947. The W orld G rows R ound M y D oor. Charles Scribners Sons, New York, NY. FARIA, C. A., TORRES, J. B., AND FARIAS, A. M. I. 2000. Reposta functional de Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) parasitando ovos de Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae): efeito da idade do hospedeiro. Anais da Entomolgica Sociadade do Brasil 29: 85 93. FARROW, S., AND MOREL, B. 2001. Continuation rights, the precautionary principle, and global change. Risk Dec Pol 6 : 145 155. FASHAM, M., AND TRUMPER, K. 2001. Review of Non Native Species Legisl ation and Guidance. Ecoscope, St Ives, UK.
227 FERNANDEZ ARHEX, V. AND CORLE Y J. C. 2003. The functional response of parasitoids and its implications for biological control. Biocontrol Sci. Technol. 4: 403 413. F ISHER, R C. 1971. Aspects of the physiol ogy of endoparasitic Hymenoptera. Biol. Rev 46: 243 278 FISHER, A., CHITOSE, A., AND GIPSON, P. S. 1994. One agencys use of risk assessment and risk communication. Risk Anal. 14 : 207 212. FISHER, A., AND CHEN, Y. 1996. Customer perceptions of agency risk communication. Risk Anal. 16: 177 184. F ISCHNOFF, B. 1990 Psychology and public policy: T ool or tool maker? Am Psychol 45: 57 63 FISCHNOFF, B. 1995. Risk perception and communication unplugged: Twenty years of process Risk Anal. 15: 137 145. FISCHNOFF, B. 2005a. Risk perception and communication, pp. 463 492 In D. Kamien [ed.], Handbook of Terrorism and Counter Terrorism. McGraw Hill, New York, NY. FISCHNOFF, B. 2005b. Decision research strategies Health Psychol 21: S9 S16. FLANDERS, S. E. 1965. On the sexuality and sex ratios of hymenopterous populations. Am erican Nat. 99: 489 494. FLEURY, F., AND BOULTREAU, M. 1993. Effects of temporary host deprivation on the reproductive potential of Tr ichogramma brassicae. Entomol. Exp. Appl. 68: 203 210. FONTA, C., PHAM DELEGUE, M. H., MARILLEAU, R., AND MASON, C. 1985. Effect of sunflower nectars on the foraging behavior of pollinating insects and quantitative and qualitative analysis of nectar glucides. Acta Oecol. Oecol. Appl. 6: 175 186. FORSSE, E., SMITH, S. M., AND BOURCHIER, R. S. 1992. Flight initiation in the egg parasitoid Trichogramma minutum : Effects of ambient temperature, mates, food and, host eggs. Entomol. Exp. Appl. 62147 154. F RANK, S A 1986. Hierarchical selection theory and sex ratios. I. General solutions for structured populations. Theor Popul Biol. 29: 312 342 F RY J. M. 1989. Natural enemy databank, 1987, pp.118 119 In J.M. Fry [ed.], A C atalogue of N atural E nemies of A rthropods D erived from R ecords in the CIBC Natural Enemy Databank CAB International, Wallingford, Oxford, UK
228 G IBSON, M. 1985 To Breathe Freely: Risk, Consent, and Air. Rowman and Allanheld, Totowa, NJ. GODFRAY, H. J. C. 1994. Host loca tion, pp. 26 81 In J.R Krebs and T. CluttonBrock [eds.], Parasitoids Behavioral and Evolutionary Ecology. Princeton University Press, Princeton, NJ. G ODIN, C ., AND B OIVIN, G. 2000. Effects of host age on parasitism and progeny alloca t ion in Trichogramm atidae. Entomol. Exp. Appl. 97: 149 160 GOLDMAN, L. 1994. Agency Progress in Risk Communication. Symposium on Risk Communication, Annapolis, MD, June 6 8 1994. GORDON D. R., AND KUBISIAK. T. L. 1998 RAPD analysis of the last population of likely Florida Keys endemic cactus. Florida Sci. 61: 203 210. GOW, H. B., AND OTWAY, H. 1990. Communicating with the public about major accidents hazard. H.B. Gow and H. Otway [eds.] Elsevier, London, UK. GRAHAM, J. D. 2001. A future for the precautionary principle? J. Risk Res. 4: 109 111. GREATHEAD, D. J. 1995. Benefits and risks of classical biological control, pp. 53 63 In H.T.M Hokkanen and J.M. Lynch [eds.], Biological Control: Benefits and Risks. Cambridge University Press, Cambridge, UK. GREATHEAD, D. J. 1997. An introduction to the FAO Code of Conduct for the import and release of exotic biological control agents. Biocontrol News Info 18: 117 118. GREENBERG, S. M., NORDLUND, D. A., AND WU, Z. 1998. Influence of rearing host on adult size an d oviposition behavior of mass produced female Trichogramma minutum Riley and Trichogramma pretiosum Riley. Biol. Control 11:43 48. G UANG, L Q ., AND O LOO, G W. 1990. Host preference studies on Trichogramma sp. nr. Mwanzai Schulten and Feijen (Hym: Tri chogrammatidae) in Kenya. Ins. Sci. APll. 11: 757 763. GURR, G. M., AND KVEDARAS, O. L. 2010. Synergizing biological control: Scope for sterile insect technique, induced plant defenses and cultural techniques to enhance natural enemy impact. Biol. Cont rol 52: 198 207. HABECK, D. H., AND BENNETT, F. D. 1990. Cactoblastis cactorum Berg (Lepidoptera: Pyralidae), a phycitine new to Florida. Florida Department of Agriculture and Consumer Services, Entomology Circular # 333. HAGVAR, E. B., AND HOFSVARY, T. 1986. Parasitism by Ephedrus cerasicola (Hymenoptera: Aphidiidae). Entomophoga 31: 337 346.
229 HAMILTON, W. D. 1967. Extraordinary sex ratio. Science 156: 477 488. HAMMOND, W. N. O, NEUENSCHWANDER, P., Y ANINEK, J. S AND H ERREN, H R 1992. Biological control in cassava: a viable crop protection package for resourcepoor farmers pp. 45 54 In R W Gibson and A Sweetmore [eds.], Proc. Semin. Crop Prot. ResourcePoor Farmers, Technical Center for Agricultural and Rural Co Operation (CTA) and Natural Res ources Institutes (NCI). H ARDY, I. C. W., D IJKSTRA, L. J., G ILLIS, J. E. M., AND L UFT, P A 1998 Patterns of sex ratio, virginity and developmental mortality in gregarious parasitoids. Biol J. Linn Soc 64: 239 270 HARDY, I. C. W., ODE, J. P., AND SIVA JOTHY, M. T. 2007. Mating behaviour, pp. 256 257 In M.A. Jervis [ed.], Insects as Natural Enemies: A pratical Perspective. Springer, Dordrecht, The Netherlands. HASSAN, S. A. 1989. Selection of suitable Trichogramm a strains to control the codling moth Cydia pomonella and the two summer fruit tortrix moths Adoxophyes orana, Pandemis heparana [Lep., Tortricidae]. Entomophaga 34: 19 27. HASSAN, S. A. 1993. The mass rearing and utilization of Trichogramma to control lepidopterous pests: A chievements and outlooks. Pestic. Sci. 37: 387 391. HASSELL, M. P., LAWTON, J. H., AND BEDDINGTON, J. R. 1977. Sigmoid functional responses by invertebrate predators and parasitoids. J. Anim. Ecol. 46: 249 262. HASSELL, M. P. 19 78. The Dynamics of ArthropodPrey Systems. Princeton University Press, Princeton, NJ. HASSELL, M. P. 2000. The Spatial and Temporal Dynamics of Host Parasitoid Interactions. Oxford Series in Ecology and Evolution. Oxford University Press, London, UK. HAYE, T., GOULET, H., MASON, P. G., AND KUHLMANN, U. 2005. Does fundamental host range match ecological host range? A retrospective case study of a Lygus plant bug parasitoid. Biol. Control 35: 55 67. HEGAZI, E. M., KHAFAGI, W. E., AND HASSAN, S. A 2000. Studies on three species of Trichogramma. IForaging behaviour for food or hosts. J. Appl. Entomol. 124: 145 149. HEIMPEL, G. E., ROSENHEIM, J. A., AND KATTARI, D. 1997. Adult feeding and lifetime reproductive success in the parasitoid Aphytis m elinus Entomol. Exp. Appl. 83: 305 315. HENNEMAN, M. L., AND MEMMOTT, J. 2001. Infiltration of a Hawaiian community by introduced biological control agents. Science 293: 1314 1316.
230 HENTZ, M. G. 1998. Development, longevity, and fecundity of Chelonus sp. nr. curvimaculatus (Hymenoptera: Braconidae), an egg larval parasitoid of pink bollworm (Lepidoptera: Gelechiidae). Environ. Entomol. 27: 443 449. H ERREN, H R AND N EUENSCHWANDER, P. 1991. Biological control of cassava pests in Africa. A nn. Rev. Entomol. 36: 257 283. HIEHATA, K., HIROSE, Y., AND KIMOTO, H. 1976. The effect of host age on the parasitism by three species of Trichogramma (Hymenoptera: Trichogrammatidae), egg parasitoids of Papilio xuthus L. (Lepidoptera: Papilionidae). Japanese J. Appl. Entomol. Zool. 20: 31 36. HIGHT, S. D., BLOSSEY, B., LAING, J., AND DECLERCK FLOATE, R. 1995. Establishment of insect biological control agents from Europe against Lythrum salicaria in North America. Environ. Entomol. 24: 967977. HIGH T, S. D., CARPENTER, J. E., BLOEM, K. A., BLOEM, S., PEMBERTON, R. W., AND STILING, P. 2002. Expanding geographical range of Cactoblastis cactorum (Lepidoptera: Pyralidae) in North America. Florida Entomol. 85: 527 529. HIGHT, S. D., CARPENTER, J. E., B LOEM, S., AND BLOEM, K. A. 2005. Developing a sterile insect release program for Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae): Effective overflooding ratios and releaserecapture field studies. Environ. Entomol. 34: 850 856. HIGHT, S. D., AND CARPENTER, J. E. 2009. Flight phenology of male Cactoblastis cactorum (Lepidoptera: Pyralidae) at different latitudes in the southeastern United States. Florida Entomol. 92: 208 216. HINZ, J. L., AND ANDOW, D. A. 1990. Host age and host selection by T richogramma nubilale. Entomophaga 35: 141 150. HODDLE, M. S. 2004. Restoring balance: Using exotic natural enemies to control invasive exotic species. Cons. Biol. 18: 38 49. HOFFMANN, M. P., WRIGHT, M. G., PITCHER, S. A., AND GARDNER, J. 2002. Inocul ative releases of Trichogramma ostrinae for suppression of Ostrinia nubilalis (European corn borer) in sweet corn: Field biology and population dynamics. Biol. Control 25: 259 258. HOLLING, C. S. 1959. The components of predation as revealed by a study of small mammal predation of the European pine sawfly. Canadian Entomol. 91: 293 320. HOMMAY, G., GERTZ, C., KIENLEN, J. C., PIZZOL, J., AND CHAVIGNY, P. 2002. Comparison between the control efficacy of Trichogramma evanescens Westwood (Hymenoptera: Tri chogrammatidae) and two Trichogramma cacoeciae Marshal strains against grape moth ( Lobesia botrana Den. and Schiff.), depending on release density. Biocontrol Sci. Technol. 12: 569 581.
231 HONDA, T., AND KAINOH, Y. 1998. Age related fecundity and learning ability of the egglarval parasitoid Acosgaster reticulatus Watanabe (Hymenoptera: Braconidae). Biol. Control 13: 177 181. HOUCK, M. A., AND STRAUSS, R. E. 1985. The comparative study of functional responses: Experimental design and statistical interpretation. Canadian Entomol. 117: 617 629. HOUSEWEART, M. W., JENNINGS, D. T., WELTY, C., AND SOUTHARD, S. G. 1983. Progeny production by Trichogramma minutum (Hymenoptera: Trichogrammatidae) utilizing eggs of Choristoneura fumiferana (Lepidoptera: Gelechi idae). Canadian Entomol. 115: 1245 1252. H OWARTH, F G. 1983. Classical biological control: Panacea or Pandoras Box? Proc. Hawaiian Entomol. Soc. 24: 239 244 HUFFAKER, C. B. 1974. Some ecological roots of pest control. Entomophaga 19: 371 389. HUGUES, R. D., WOOLCOK, L. T., AND HUGHES, M. A. 1992. Laboratory evaluation of parasitic Hymenoptera used in attempts to biologically control aphid pests of crops in Australia. Entomol. Exp. Appl. 63: 177 185. HUNT, E. J., KULHMANN, U., SHEPPARD, A., Q IN, T. K., BARRATT, B. I. P., HARRISSON, L., MASON, P.G., PARKER, D., FLANDERS, R. V., AND GOOLSBY, J. 2008. Review of invertebrate biological control agent regulation in Australia, New Zealand, Canada and the USA: Recommendations for harmonized European system. J Applied Entomol. 132: 89 123. IPPC (INTERNATIONAL PLANT PROTECTION CONVENTION). 1997. Code of Conduct for the Import and Release of Exotic Biological Control Agents. International Standard for Phytosanitary Measures # 3. Food and Agriculture Organization of the United Nations, Rome, Italy. IPPC (INTERNATIONAL PLANT PROTECTION CONVENTION). 2004. Guidelines for Pest Risk Analysis. International Standard for Phytosanitary Measures # 2. Food and Agriculture Organization of the United Nations, Rome, Italy. IPPC (INTERNATIONAL PLANT PROTECTION CONVENTION). 2005. Guidelines for the Export, Shipment, Import and Release of Biological Control Agents and other Beneficial Organisms. International Standard for Phytosanitary Measures # 3. Food and Agriculture Organization of the United Nations, Rome, Italy. IPPC (INTERNATIONAL PLANT PROTECTION CONVENTION). 2007. Pest Risk Analysis for Quarantine Pest including Analysis of Environmental Risks and Living Modified Organisms. International Standard for Phytosanitary Measures # 11. Food and Agriculture Organization of the United Nations, Rome, Italy.
232 IVES, A. R., AND SETTLE, W. H. 1996. The failure of a parasitoid to persist with a superabundant host: The importance of the numerical response. Oik os 75: 269 278. I VES, A R S CHOOLER, S S J AGAR, V J. K NUTESON, S E GRBIC, M S ETTLE, W H 1999 Variability and parasitoid foraging efficiency: A case study of pea aphids and Aphidius ervi Am erican Nat 154: 652673. IRA (IMPORT RISK ANALY SIS). 2007. Import Risk Analysis Handbook. [Online] http://www.daff.gov.au/ba/ira/process handbook Last accessed 03/24/2011. JAEGER C. J., R ENN O., R OSA E. A., AND W EBLER T. 2001. Risk, Uncertainty, and Rational Action. Earthscan, London, UK. JASANOFF, S. 1989. Norms for evaluating regulatory science. Risk Anal. 9: 271 273. JAYANTH, K. P., MOHANDAS, S., ASOKAN, R., GANGA, AND VISALAKSHY, P. N. 2003. Parthenium pollen induced f eeding by Zygogramma bicolorata (Coleoptera; Chrysomelidae) on sunflower ( Helianthus annuus ) (Compositae). Bull. Entomol. Res. 83: 595 589. JOHNSON, D. M., AND STILING, P. 1996. Host specificity of Cactoblastis cactorum Berg, an exotic Opuntia feeding m oth, in Florida. Environ. Entomol. 25: 743 748. JOHNSON, D. M., AND STILING, P. 1998. Distribution and dispersal of Cactoblastis cactorum (Lepidoptera: Pyralidae), an exotic Opuntia feeding moth in Florida. Florida Entomol. 81: 12 22. JUNGERMANN, H., S CHUTZ, H., AND THURING, M. 1988 Mental models in risk assessment: Informing people about drugs. Risk Anal. 8: 147 155. JULIEN, M. H. 1992. Biological Control of Weeds: A World Catalogue of Agents and Their Target Weeds, 3rd Ed. CAB International, W allingford, UK. JULIEN, M. H. AND G RIFFITHS M. W. 1998. Biological C ontrol of W eeds. A W orld C atalogue of A gents and T heir T arget W eeds. 4th Ed CABI Publishing Wallingford, UK KAIRO, M. T. K. 2005. Hunger, poverty, and protection of biodiversity: Opportunities and challenges for biological control, pp. 228 236 In Hoddle, R.G. (Compiler) Proceedings of the 2nd International Symposium on Biological C ontrol of Arthropods, Davos, Switzerland, 12 16 September 2005. US Department of Agric ulture, Forest Service, Morgantown, WV. KALYEBI, A., OVERHOLT, W. A., SCHULTHESS, F., MUEKE, J. M., HASSAN, S. A., AND SITHANANTHAM, S. 2005. Functional response of six indigenous trichogrammatid egg parasitoids (Hymenoptera: Trichogrammatidae) in Kenya: Influence of temperature and humidity. Biol. Control 32: 164 171.
233 KING, B. H., GRIMM, K. M., AND RENO, H. E. 2000. Effects of mating on female locomotor activity in the parasitoid wasp Nasonia vitripennis (Hymenoptera: Pteromalidae). Environ. Entomol 29: 927 933. K NIPLING E. F. 1992. Principles of Insect Parasitism Analyzed From New Perspectives: Practical Implications for Regulating Insect Populations by Biological Means. U.S. Department of Agriculture. Agriculture Handbook No 693. Government Printing Office, Washington, DC. KOCH, F. H., YEMSHANOV, D., MCKENNEY, D. W, AND SMITH, W. D. 2009. Evaluating critical uncertainty thresholds in a spatial model of forest pest invasion risk. Risk Anal. 29: 12271241. KOGAN, M., GELING, D. AND MADDOX J. V. 1999. Enhancing biological control in annual agricultural environments, pp. 789 818 In T.S. Bellows and T. W. Fisher [ eds.], Handbook of Biological Control. Academic Press, San Diego CA KOPTUR, S., AND TRUONG, N. 1998. Facultative ant plant interactions: Nectar sugar preferences of introduced pest ant species in South Florida. Biotropica 30: 179 189. K UBASEK, N K AND S ILVERMAN, G S 2005. Environmental Law, 5th ed. Pearson Education Inc. Upper Saddle R iver, NJ KUHLMANN, U. MASON, P. G., HINZ, H. L., BLOSSEY, B., DE CLERCK FLOATE, R. A., DOSDALL, L. M., MCCAFFREY, J. P., SCHWARLAENDER, M., OLFERT, O., BRODEUR, J., GASSMANN, A., MCCLAY, A. S., AND WIEDENMANN, R. N. 2006. Avoiding conflicts between insect and weed biological control: Selection of nontarget species to assess host specificity of cabbage seedpod weevil parasitoids. J. Appl. Entomol. 130: 129 141. KUMAR, N., KUMAR, A., AND TRIPATHI, C. P. M. 1994. Functional response of Campoletis chlori dae Uchida (Hymenoptera: Ichneumonidae), a parasitoid of Heliotis armigera (H bner) (Lepidoptera: Noctuidae) in an enclosed experimental system. Biol. Agr. Hort. 10: 287 295. LAETEMIA, J. A., LAING, J. E., AND CORRIGAN, J. E. 1995. Effects of adult nutr ition on longevity, fecundity, and offspring sex ratio of Trichogramma minutum Riley (Hymenoptera: Trichogrammatidae). Canadian Entomol. 127: 245 254. LANDIS, D. A., SEBOLT, D. C., HAAS, M. J., AND KLEPINGER, M. 2003. Establishment and impact of Galeruc ella calmariensis L. (Coleoptera: Chrysomelidae) on Lythrum salicaria L. and associated plant communities in Michigan. Biol. Control 28: 78 91. L AROCHE, F B ., AND FERRITER, A P. 1993. The rate of expansion of Melaleuca in south Florida. J. Aquat. Plan t Manage. 30: 62 65.
234 LAUMANN, R. A., MORAES, M. C. B., PAREJA, M., ALARCAO, G. C., BOTELHO, A. C., MAIA, A. H. N., LEONARDECZ, E., AND BORGES, M. 2008. Comparative biology and functional response of Trissolcus spp. (Hymenoptera: Scelionidae) and implicat ions for stink bugs (Hemiptera: Pentatomidae) biological control. Biol. Control 44: 32 41. LAVE, T. R., AND LAVE, L. B. 1991. Public perception of the risks of floods: implication for communication. Risk Anal. 11: 255 267. LI, L Y 1994. Worldwide use of Trichogramma for biological control on different crops: A survey, pp. 37 53 In E. Wajnberg and S. A. Hassan [eds.] Biological Control with Egg Parasitoids CAB International, Wallingford, UK LOGARZO, G., VARONE, L., AND BRIANO, J. 200 9 Cactus Moth, In Annual Report 2009 South American Biological Control Laboratory, United States Department of Agriculture, Agricultural Research Service, USDA ARS, Hurlingham, Argentina. [Online] http://www.ars.usda.gov/SP2UserFiles/Place/02110000/CompleteAnnualReport20 09.pdf Last accessed 04/05/11. LOOMANS, A. M. J. 2007. Regulation of biological control agents in Europe: Review and recommendations in its pursuit of a harmonized system. Report REBCA (Regulation of Biological Control Agents). [Online] http://www.rebeca net.de/downloads/Regulation%20of%2 0Beneficials%20in%20Europe.pdf Last accessed 04/01/2011. LOPES, C. SPATANO, T., LAPCHIN, L., AND ARDITI, R. 2008. Optimal release strategies for biological control of aphids in melon greenhouses. Biol. Control 48: 12 21. LOUDA, S M P EMBERTON, R W J OHNSON, M T AND F OLLETT, P A. 2003. Non target effects The Achilles heel of biological control? Retrospective analyses to reduce risk associated with biocontrol introductions. Ann. Rev. Entomol. 48: 365 396. L UCK, R F ., STOUTHAMER, R N UNNEY, L 1992. Sex determination and sex ratio patterns in parasitic Hymenoptera, pp. 442 476 In D.L. Wrench and M. Ebbert [ eds .], Evolution and D iversity of S ex R atio in H aplodiploid I nsects and Mites. Chapman & Hall, New York, NY. LUNDGREN, R., AND MCMAKIN, A. 2004. Approaches to communicating risk, pp. 1328 In R. Lundgren and A. McMakin [eds.], Risk Communication: A Handbook for Communicating Environmental, Safety, and Health Risks. Battelle Press, Columbus, OH. MAHARIK, M., AND FISCHNOFF, B. 1 992. The risks of using nuclear energy sources in space: Some lay activists perceptions. Risk Anal. 12: 383 392.
235 MANN, J. 1970. Cacti Naturalized in Australia and Their Control. Department of Lands, Queensland, Australia. MANSFIELD, S., AND MILLS, N J. 2004. A comparison of methodologies for the assessment of host preference of the gregarious egg parasitoid Trichogramma platneri Biol. Control 29: 332 340. MARTEL, V., AND BOIVIN, G. 2004. Premating dispersion in the egg parasitoid Trichogramma (Hymenoptera: Trichogrammatidae). Pop. Ecol. 33: 855 859. MARTIN, P. B., LINGREN, P. D., GREEN, G. L., AND RIDGWAY, R. L. 1976. Parasitization of two species of Plusiinae and Heliotis spp. after releases of Trichogramma pretiosum in seven crops. Environ. Entomol. 5: 991 995. MASON, P. G., FLANDERS, R. G., AND ARRENDONDO BERNAL, H. A. 2005. How can legislation facilitate the use of biological control of arthropods in North America? pp. 701 714 In Hoddle, R.G. (Compiler) Proceedings of the 2nd International Symposium on Biological C ontrol of Arthropods, Davos, Switzerland, 12 16 September 2005. US Department of Agriculture, Forest Service, Morgantown, WV. MCEVOY, P. B. 1996. Host specificity and biological pest control. BioScience 46: 401 4 05. MCFAYDEN, R. E. 1985. Larval characteristics of Cactoblastis spp. (Lepidoptera: Pyralidae) and the selection of species for biological control of prickly pears ( Opuntia spp.). Bull Entomol Res 75: 159 168. MCFAYDEN, R. E. 1998 Biological contr ol of weeds. Ann. Rev. Entomol. 43: 369 393. M ESSING, R H 2005 Hawaii as a role model for comprehensive U.S. b iocontrol legislation: T he best and the worst of it pp. 686 691 In Hoddle, R.G. (Compiler) Proceedings of the 2nd International Symposium on Biological control of Arthropods, Davos, Switzerland, 12 16 September 2005. US Department of Agriculture, Forest Service, Morgantown, WV. MESSING, R. H., ROITBERG, B. D., AND BRODEUR, J. 2006. Measuring and predicting indirect impacts of biological control, competition, displacement and secondary interactions, pp. 64 77 In F. Bigler, D. Babendreier, and U. Kuhlmann [eds.], Environmental Impact of Inverterbrates for Biological Control of Arthropods Methods and Risk Assessment. CABI Publishing, Wallingford, UK. MESSING, R. H., AND WRIGHT, M. G. 2006. Biological control of invasive species: Solution or pollution? Front. Ecol. Environ. 4: 132140. M EYER M. A., AND B OOKER J. M. 1990 Elicit ing and A nalyzing E xpert Judgment: A P ractical G uide. Nuclear Regulatory Commission, Washington, DC.
236 MILLS, N. J., AND KUHLMANN, U. 2000. The relationship between egg load and fecundity among Trichogramma parasitoids. Ecol. Entomol. 25: 315 324. MILLS N. J., AND KUHLMANN, U. 2004. Oviposition behavior of Trichogramma platneri Nagarkatti and Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) in patches of single and clustered host eggs Biol. Control 30: 42 51. MILLS, N. J., AND LACAN, I. 2004. Ratio dependence in the functional response of insect parasitoids: Evidence from Trichogramma minutum foraging for eggs in small host patches. Ecol. Entomol. 29: 208 216. MIURA, K., AND KOBAYASHI, M. 1998. Effects of host age on the parasitism by Trichogramma chilonis Ishii (Hymenoptera: Trichogrammatidae), an egg parasitoid of the diammonback moth. Appl. Entomol. Zool. 33: 219 222. M ONJE J. C., Z EBITZ C. P. W. AND O HNESORGE, B. 1999. Host and host age preference of Trichogramma galloi and T. pretiosum (Hymenoptera: Trichogrammatidae) reared on different hosts. J. Econ. Entomol. 92: 97 103. MONRO, J. 1967. The exploitation and conservation of resources by populations of insects. J. Animal Ecol. 36: 531 547. MORAN, V. C., AND ZIMMERMANN, H. G. 1984. The biological control of cactus weeds: Achievements and prospects. Biocontrol News Infor 5: 297 320. MORGAN, M. G., FISCHNOFF, B., BOSTROM. A., AND ATMAN, C. J. 2002. Risk Communication: A Mental Models Approach. Cambridge University P ress, New York, NY. MORRISON, R. K. 1985. Trichogramma spp. Vol. I, pp. 413 417 In P. Singh and R. F. Moore [eds.], Handbook of Insect Rearing, Vol. 1. Elsevier, Amsterdam, The Netherlands. MORTON, J. F. 1966. The cajeput treea boon and an affliction. Econ. Bot. 20: 31 39. MUNYANEZA, J., AND OBRYCKI, J. J. 1997. Functional response of Coleomegilla maculata (Coleoptera: Coccinelidae) to Colorado potato beetle eggs (Coleoptera: Chrysomelidae). Biol. Control 8: 215 224. MYERS, J. H., MONRO, J., AND MURRAY, N. 1981. Egg clumping, host plant selection, and population regulation in Cactoblastis cactorum (Lepidoptera). Oecolgia 51: 7 13.
237 NAPPO (North American Plant Protection Organization). 2002. Guidelines for Petition for Release of Exotic Entomophagous Agents for the Biological Control of Pests (RSPM # 12). [Online] http://www.nappo.org/Standards/Standards(all)/RSPM12Rev201008e.pdf Last accessed 04/05/11. NAPPO (North American Plant Protection Organization). 2006. Official Pest Reports: Detection of an outbreak of cactus moth ( Cactoblastis cactorum ) in Isla Mujeres, Quintana Roo, Mexico. [Online] www.pestalert.org Last accessed 12/27/10. NAPPO (North American Plant Protection Organization). 2008. Official Pest Reports: Eradication of cactus moth ( Cactoblastis cactorum Berg ) outbreak in Isla Mujeres, Quintana Roo, Mexico. [Online] www.pestalert.org Last accessed 12/27/10. NAPPO (North American Plant Protection Organization). 2009. Official Pest Reports: Detection and eradication of a cactus moth ( Cactoblastis cactorum Berg) outbreak in Isla Contoy, municipality of Isla Mujeres, Quintana Roo, Mexico. [Online] www.pestalert.org Last accessed 12/27/10. NATIONAL RESEARCH COUNCIL. 1996. National Science Education Standards. National Ac ademy Press, Washington, D C. NEPA (NATIONAL ENVIRONMENTAL POLICY ACT). 1970. EA and EIS component [Online] http://www.epa.gov/compliance/basics/nepa.html Last accessed 12/27/10. NORGAARD, R B. 2006. Economics of the cassava mealybug [ Phaenacoccus Manihoti ; Hom.: Pseudococcidae] biological control program in Africa. BioControl 33: 1 3. N OYES J. S. 1998. Catalogue of Chalcidoidea of the World. CDROM Series, ETI, Amsterdam, Netherlands NOYES, J. S., AND V ALENTINE, E. W. 1989. Chalcidoidea (Insecta: Hymenoptera) I ntroduction, and review of genera in smaller families. Fauna New Zealand 18:1 91. NUNNEY, L. 1985. Femalebiased sex rat ios: Individual or group selection? Evolution 39: 349 361 NURINDAH, C. B. W., AND GORDH, G. 1999. Effects of physiological condition and experience on oviposition behavior of Trichogramma australicum Girault (Hymoneptera: Trichogrammatidae) on eggs of Helicoverpa armigera Hbner (Lepidoptera: Noctuidae). Australian J. Entomol. 38: 104 114. OECD (ORGANIZATION FOR ECONOMIC COOPERATION AND DEVELOPMENT). 2004. Guidance for Information Requirements for Regulation of Invertebrates as Biological Control Agents. [Online] http: //www.oecd.org Last accessed 03/24/11.
238 OLIVEIRA, H. N., ZANUNCIO, J. C., PRATISSOLI, D., AND PICANO, M. C. 2003. Biological characteristics of Trichogramma maxacalii (Hymenoptera: Trichogrammatidae) on eggs of Anagasta kuehniella (Lepidoptera: Pyralidae). Braz illian J. Biol. 63: 647 653. OLKOWSKI, W., AND ZHANG, A. 1990. Trichogramma modern day frontier in biological control. IPM Practitioner 12: 1 15. ORR, D. B., GARCIA SALAZAR, C., AND LANDIS, D. A. 2000. Trichogramma nont arget impacts: A method for biological control risk assessment, pp. 111 125 In P.A. Follet and J.J. Duan [eds.], Nontarget Effects of Biological Control. Kluwer Academic Publishers, Norwell, MA. PAGE, T. 1978. A generic view of toxic chemicals and simi lar risks Ecol. Law Quat 7: 207 244. PAK, G. A. 1986. Behavioural variations among strains of Trichogramma spp.: A review of the literature on host age selection. J. Appl. Entomol. 101: 55 64. PAK, G. A., AND OATMAN, E. R. 1982. Comparative life ta ble, behavior and competition studies of Trichogramma brevicapillum and T. pretiosum Entomol. Exp. Appl. 32: 68 79. PANDEY, K. P., SINGH, R., AND TRIPATHI, C. P. M. 1984. Functional response of Diaeretiella rapae (McIntosh) (Hymenoptera: Aphidiidae), a parasitoid of the mustard aphid Lipaphis erymisi Kalt. (Homoptera: Aphididae). Z. Angew. Entomol. 98: 321 327. PARAISO, O., HIGHT, S. D., KAIRO, M. T. K., AND BLOEM, S. 2011. E gg parasitoids attacking Cactoblastis cactorum (L epidoptera : P yralidae) in N orth F lorida. Florida Entomol.94: 91 90. PARKER, F. D., AND PINNELL, R. E. 1973 Effect on food consumption of the imported cabbageworm when parasitized by two species of Apanteles Environ. Entomol. 2: 216 219. PARRA, J. R. P., ZUCCHI, R. A., AND SILVEIRA NETO, S. 1987. Biological control of pests through egg parasitoids of the genera Trichogramma and/or Trichogrammatoidea. Mem. Inst. Oswaldo Cruz 82: 153 160. PEMBERTON, R. W. 1995. Cactoblastis cactorum (Le pidoptera: Pyralidae) in the United States, an immigrant biological control agent or an introduction of the nursery industry? Am erican Entomol. 41: 230 232. PEMBERTON, R. W. 2000. Predictable risk to native plants in weed biological control. Oecologia 125: 489 494.
239 PEMBERTON, R. W., AND CORDO, H. 2001a. Potential and risk of biological control of Cactoblastis cactorum (Lepidoptera: Pyralidae) in North America. Florida Entomol. 84: 513 526. PEMBERTON, R. W., AND CORDO, H. 2001b. Nosema (Microsporidi a: Nosematidae) species as potential biological control agents of Cactoblastis cactorum (Lepidoptera: Pyralidae): S urveys for the microsporidia in Argentina and South Africa. Florida Entomol. 84: 527530. PHILIP, M. M., ORR, D. B., AND HAIN, F. P. 2005. Evaluation of biological and biorational control tactics for suppression of nantucket pine tip moth damage in Virginia pine Christmas trees. J. Econ. Entomol. 98: 409 414. P IMENTEL, D L ACH, L Z UNIGA, R AND M ORRISON, D. 1999. Environmental and economic costs of non indigenous species in the United States. BioScience 50: 53 65. PIMENTEL, D., ZUNIGA, R., AND MORRISON, D. 2005. Update on the environmental and economic costs associated with alien invasive species in the United States. Ecol. Econ 5 2: 273 288. PINTO, J. D. 1998. Systematics of the North American species of Trichogramma Westwood (Hymenoptera: Trichogrammatidae). Mem. Entomol. Soc. Washington 22:1 287. P INTO J. D. 1999. Systematics of the North American species of Trichogramma Westwood (Hymenoptera: Trichogrammatidae). Mem. Entomolo Soc Wash ington 22: 140 PINTO, J. D., OATMAN, E. R., AND PLATNER, G. R. 1986. Trichogramma pretiosum and a new cryptic species occurring sympatrically in Southwestern North America (Hymenoptera: Trichogrammatidae). Ann. Entomol. Soc. America 79: 1019 1028. PINTO, J. D. AND STOUTHAMER, R. 1994 Systematics of the Trichogrammatidae with emphasis on Trichogramma, Chap. 1, pp. 1 36 In E. Wajnberg and A. Hassan [eds.], Biological Control with Eggs Parasitoids. CAB International, Wallingford, UK. P INTUREAU, B., P ETINON, S., AND N ARDON C. 199 9. Possible function of substances excreted by Trichogramma and darkening of their hosts. Bul l Soc Zool France 124 : 261 269 PLUKE, R. W. H., AND LEIBEE, G. L. 2006. Host preferences of Trichogramma pretiosum and the influence of prior ovipositional experience on the parasitism of Plutella xylostella and Pseudoplusia includens eggs. BioControl 51: 569 583.
240 PODOLER, H., ROSEN, D., SHARONI, M. 1978. Ovipositional responses to host density in Aphytis holoxanthus (Hymenoptera: Aphelinidae), an efficient gregarious parasite. Ecol. Entomol. 3: 305 311. POMPANON, F., FOUILLET, P., AND BOULETREAU, M. 1999. Physiological and genetic factors as sources of variation in locomotion and activity rhythm in a parasitoid wasp ( Trichogramma brassicae). Physiol. Entomol. 24: 346 357. PPA ( PLANT PROTECTION ACT ) 2000. Plant Protection. Act Commerce and T rade, E xports and I mports, 7 USC 7701 note, Public Law 106 224. [ O nline] www.aphis.usda.gov/brs/pdf/PlantProtAct2000.pdf Last accessed 01/12/ 20 1 1 PPQS (PLANT PROTECTION, QUARANTINE AND STORAGE). 2006. Guidelines for Regulating Export, Import and Release of Biological Control Agents and Other Beneficial Organisms. [Online] http://plantquarantineindia.org/pdffiles/guidlelinesforbeneficial%20organisms_7AU G2006.pdf Last accessed 09/01/2007. QUERINO, R. B., AND ZUCCHI, R. A. 2003. New species of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) associated with lepidopterous eggs in Brazil. Zootaxa 163: 1 10. RABINOVICH, J. E. 1970 Population dynamics of Telenomus fariai (Hymenoptera: Scelionidae) parasite of Chagas disease vectors. IV. Effects of female size on progeny. J. Med. Entomol. 7: 561 567 RAFOSS, T. 2003. Spatial stochastic simulation offers potential as a quantitative method for pest risk analysis. Risk Anal. 23: 651661. RAIFFA, H. 1968. Decision Analysis: Introductory Lectures on Choice under Certainty. AddisonWesley, Reading, M A. RAJAPAKSE, R. H. S. 1992. Effect of host age, parasitoid age, and temperature on interspecific competition between Chelonus insularis Cresso, Cotesia marginventris Cresson and Microplitis manila Ashmead. Insect Sci. Appl. 13:87 94. RAM, A., AND IRULANDI, V. 1989. Influence of host egg on biology of Trichogramma exiguum Pinto, Platner & Oatman (Hym.: Trichogrammatidae). Indian J. Entomol. 51:361 365. REBECA (REGULATION OF BIOLOGICAL CONTROL AGENTS). 2006. General Topics, pp. 7 19 In Final Repor t Regulation of Biological Control Agents Specific Support Action, Sustainable Management of Europes Natural Resources. REBECA (REGULATION OF BIOLOGICAL CONTROL AGENTS). 2011. Projects Description. [Online] http://www.rebeca net.de/ Last accessed 03/24/11.
241 RENDON, P., S IVINSKI J., H OLLER T., B LOEM K., L OPEZ M., M ARTINEZ A., AND A LUJA, M. 2006. The effects of sterile males and two braconid parasitoids, Fopius arisanus (Sonan) and Diachasmimorpha krausii (Fullaway) (Hymenoptera), on caged populations of Mediterranean fruit flies, Ceratitis capitata (Wied.) (Diptera: Tephritidae) at various sites in Guatemala. Biol. Control 36: 224 231. R OBERTSON H. G. 1984. Egg predation by ants as a partial explanatio n of the difference in performance of Cactoblastis cactorum on cactus weeds in South Africa and Australia, pp. 83 88 In E.S. Delfosse [ed.], Proc. VI Symp. Biol. Contr. Weeds, 19 25 August 1984, Vancouver, Canada. Agriculture Canada, Ottawa. ROBERTSON, H. G. 1987. Oviposition site selection in Cactoblastis cactorum (Lepidoptera): Constraints and compromises. Oecologia 73: 601 608. ROBERTSON, H. G. 1989. Seasonal temperature effects on fecundity of Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae): Differences between South Africa and Australia. J. Entomol. Soc. South Africa 52: 71 80. ROBERTSON, H. G., AND HOFFMANN, J. H. 1989. Mortality and life tables of Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) compared on two host plant species. Bull. Entomol. Res. 79: 7 17. ROMEIS, J., BARBENDREIER, D., WACKERS, F. L., AND SHANOWER, T. G. 2005. Habitat and plant specificity of Trichogramma egg parasitoids underlying mechanisms and implications. Basic Appl. Ecol. 6: 215 236. ROMEIS, J. T., SHA NOWER, G., AND ZEBITZ, P. W. 1997. Volatile plant infochemicals mediate plant preference of Trichogramma chilonis J. Chem. Ecol. 23: 2455 2465. ROMEIS, J. T., SHANOWER, G., AND ZEBITZ, P. W. 1999. Trichogramma egg parasitism of Helicoverpa armigera o n pigeonpea and sorghum in southern India. Entomol. Exp. Appl. 90: 69 81. ROWE, G., MARSH, R., AND FREWER, L. J. 2004. Evaluation of a deliberative conference in science. Tech Hum Val 19: 88 121. R UBERSON, J. R ., AND KRING, T. J. 1993. Parasitism of developing eggs by Trichogramma pretiosum (Hym: Trichogrammatidae): H ost age preference and suitability. Biol. Control 3: 39 46 RUESINK, J. L., PARKER, I.M., GROOM, M. J., AND KAREIVA, P. M. 1995. Reducing the risks of nonindigenous species introductions. Bioscience 45: 465 477. SANTOS, S., AND CHESS, C. 2003. Evaluating citizen a dvisory b oards: The i mportance of t heory and p articipant b ased criteria and p ractical i mplications. Risk Ana l. 23: 269 279.
242 SAS INSTITUTE. 1999. SAS/STAT Users Guide, Version 8.01. SAS Institute, Inc., Cary, NC. SCOLES, J., CUDA, J. P., AND OVERHOLT, W. A. 2008. How scientists obtain approval to release organisms for classical biological control of invasiv e weeds. [Online] http://edis.ifas.ufl.edu/pdffiles/IN/IN60700.pdf Last accessed 04/01/2011. S ELIN S., AND C HAVEZ, D 1995. Developing a collaborative model for environmental planning and management. J. Env iron. Manage. 19: 189 195. SENGUE, P. M. 1992. The Fifth Discipline: The Art and Practice of the Learning Organization. Doubleday Inc., New York, NY. SEQUEIRA, R., AND MACKAUER, M. 1988. Effects of parasitism by Praon pequdorum on age specific fecundity and population growth of the pea aphid, Acyrthosiphon pisum Entomol. Exp. Appl. 48: 179 185. SEQUEIRA, R., AND MACKAUER, M. 1992. Covariance of ad ult size and development time in the parasitoid wasp Aphidius ervi in relation to the size of host Acrytosiphon pisum Evol. Ecol. 6: 34 44. SEQUEIRA, R., AND MACKAUER, M. 1994. Variation in selected life history parameters of the parasitoid wasp, Aphid ius ervi : Influence of host developmental stage. Entomol. Exp. Appl. 71: 15 22. SERBESOFF KING, K. 2003. Melaleuca in Florida: A literature review on the taxonomy, distribution, biology, ecology, economic importance, and control measures. J. Aquat. Pl ant Manage. 41: 98112. SCHMIDT, J. M., AND SMITH, J. J. B. 1985. The mechanism by which the parasitoid wasp Trichogramma minutum responds to host clusters. Entomol. Exp. Appl. 39 : 287 294. SCHMIDT, J. M., AND SMITH, J. J. B. 1986. Measurement of hos t curvature by the parasitoid wasp Trichogramma minutum and its effect on host examination and progeny allocation. J. Exp. Biol. 129: 151 164. SCHMIDT, J. M., AND SMITH, J. J. B. 1989. Host examination walk and ovipositing site selection of Trichogramm a minutum : S tudies on spherical hosts. J. Ins. Behav. 2: 143 171. SHANNON, C .E. 1948. A Mathematical Theory of Communication. Bell System Technical Journal, Bell Labs, Murray Hill, NJ. S HEPPARD, A. W., HILL R., D E C LERCK F LOATE R. A., M CCLAY A., OLC KERS T., Q UIMBY Jr., P. C., AND Z IMMERMANN H. G. 2003. A global review of risk benefit cost analysis for the introduction of classical biological control agents against weeds: A crisis in the making? Biocontrol News Info. 24 : 91108.
243 SIDDIQUI, I. R. 1970. The sugars of honey. Adv. Carb. Chem. Bi. 25: 285 309. SIMBERLOFF, D. 1996. Impacts of Introduced Species in the United States. [Online] http://www.gcrio.org/CONSEQUENCES/vol2 no2/article2.html Last accessed 04/01/2011. SIMBERLOFF, D. 2005. The politics of assessing risk for biological invasions: The USA as a case study. Trends Ecol. Evol. 20: 216 222. S IMBERLOFF, D AND S TILING, P. 1996. How risky is biological control? Ecol. 77: 1965 1974 SIMMONDS, F. J., AND BENNETT, F. D. 1966. Biological control of Opuntia spp. by Cactoblastis cactorum in the Leeward Islands (West Indies). Entomophaga 11: 183 189. SIMONSEN, T. J., BROWN, R. L., AND SPERLING, F. A. 2008. Tracin g an invasion: Phylogeography of Cactoblastis cactorum (Lepidoptera: Pyralidae) in the United States based on Mitochondrial DNA. Ann. Entomol. Soc. America 101: 899 905. SITHANANTHAM, S., ABERA, T. H., BAUMGARTNER, J., HASSAN, S. A., LOHR, B., MONJE, J. C ., OVERHOLT, W. A., PAUL, A. V. N., FANG HAO, W., AND ZEBITZ, C. P. W. 2001. Egg parasitoids for augmentative biological control of lepidopteran vegetable pests in Africa: Research status and needs. Insect Sci. Appl. 21: 189 205. SIVINSKI, J. M. 1996. The past and potential of biological control of fruit flies, pp. 369 377 In J.B.A. McPheron and G.J. Steck [eds.], Fruit Fly Pests, A World Assessment of their Biology and Management. St. Lucie Press, Delray Beach, FL. SMITH, S. M. 1996. Biological control with Trichogramma advances, successes, and potential of their use. Ann. Rev. Entomol. 41: 375 406. SOKAL, R. R., AND ROHLF, F. J. 1981. Kruskal Wallis pp. 429 432 In J. Wilson and S. Cotter [eds], Biometry. WH. Freeman and Co. New York, NY. SOLOMON, M. E. 1949. The natural control of animal populations. J. Anim. Ecol.18: 1 35. STILING, P. D. 2002. Potential nontarget effects of a biological control agent, prickly pear moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), in North Ame rica, and possible management actions. Biol. Invasions 4: 273 281. STILING, P. D., MOON, D., AND GORDON, D. 2004. Endangered cactus restoration: Mitigating the nontarget effects of biological control agent ( Cactoblastis cactorum ) in Florida. Rest. Eco l 12: 605 610.
244 STRAND, M. R., AND VINSON, S. B. 1984. Facultative hyperparasitism by the egg parasitoid Trichogramma pretiosum (Hymenoptera: Trichogrammatidae). Ann. Entomol. Soc. America 77: 679 686. STRINGER, L. C., AND REED, M. S. 2007. Land degradation assessment in southern Africa: Integrating local and scientific knowledge bases Land Degrad. Dev 18: 99 116. SUCKLING, D. M., BARRINGTON, A. M. CHHAGAN, A., STEPHENS, A. E. A. BURNIP, G. M., CHARLES, J. G., AND WEE, S. L. 2007. Eradication of the Australian painted apple moth Teia anartoides in New Zealand: Trapping, inherited sterility, and male competitiveness, pp. 603 615 In M.J.B. Vreysen, A.S. Robinson and J. Hendrichs [eds.], AreaWide Control of Insect Pests. Springer, Dordrecht, The Ne therlands. SUH, C. P., ORR, D. B., AND VAN DUYN, J. W. 2000. Trichogramma releases in North Carolina cotton: Why releases fail to suppress heliothine pests. J. Econ. Entomol. 93: 1137 1145. S UZUKI, Y AND H IEHATA, K 1985. Mating systems and sex rati os in the egg parasitoids, Trichogramma dendrolimi and T. papilionis (Hymenoptera: Trichogrammatidae). Anim Behav 33: 1223 1227. SUZUKI, Y., TSUJI, H., AND SASAKAWA, M. 1984. Sex allocation and effects of superparasitism on secondary sex ratios in the gregarious parasitoid, Trichogramma chilonis (Hymenoptera: Trichogram matidae). Anim. Behav. 32: 478 484. TAGAWA, J., YOSHIDA, C., HASHIMOTO, T., AND SUDARE, A. 1987. Effects of mating on the oviposition behaviour of the parasitic wasp, Apanteles glomeratus L. (Hymenoptera: Braconidae). J. Ethol. 5: 37 41. TAKADA, Y., KA WAMURA, S., AND TANAKA, T. 2000. Biological characteristics: Growth and development of the egg parasitoid Trichogramma dendrolimi (Hymenoptera: Trichogrammatidae) on the cabbage armyworm Mamestra brassicae (Lepidoptera: Noctuidae). Appl. Entomol. Zool. 35: 369 379. TAYLOR, P. D. 1981. Intra sex and Inter sex sibling interactions as sex ratio determinants. Nature 291: 64 66. T HOMAS M. B. AND W ILLIS, A. J. 1998. Biocontrol R isky but necessary? Trends Ecol Evol 13: 325 329. THOMSON, M. S., AND STINNER, R. E. 1989. Trichogramma spp. (Hymenoptera: Trichogrammatidae): Field hosts and multiple parasitism in North Carolina. J. Entomol. Sci. 24: 232 240.
245 USBC (U.S. BUREAU OF CENSUS). 2001. Statistical Abs tract of the United States 2001. U.S Bureau of the Census, U.S. Government Printing Office, Washington, DC. VAN ALEBEEK, F. A. N., KONING, C. M., DE KORTE, E. A. P., AND VAN HUIS, A. 1996. Eggs limited, functional response of Uscana lariophaga, egg parasitoid of bruchid beetle pests in st ored cowpea. Entomol. Exp. Appl. 81: 215 225. VAN DEN ASSEM, J., AND VISSER, J. 1976. Aspects of sexual receptivity in female Nasonia vitripennis Biol. Comp. 1: 37 56. VAN DEN ASSEM, J., BOSCH, H. A. J., AND IN DEN PROOY, E. 1982. Melittobia courtsh ip behaviour: A comparative study of the evaluation of a display. Neth erlands J. Zool. 32: 427 471. VAN DRIESCHE, R. G., AND HODDLE, M. 1997. Should arthropod parasitoids and predators be subject to host range testing when used as biological control age nts? Agric. Hum. Val. 14: 211 226. V AN D RIESCHE R. G. AND R EARDON, R. 2004. Assessing H ost R anges for P arasitoids and P redators used for C lassical B iological C ontrol: A guide to B est P ractice. United States Department of Agriculture Forest Health Technology Enterprise Team, Morgantown, W V VAN LENTEREN, J. C. 2000. Success in biological control of arthropods by augmentation of natural enemies, pp. 77 103 In G. Gurr and S. Wratten [eds.], Biological Control: Measures of Success. Kluwer Academic P ublishers, Hingham, MA VAN LENTEREN, J. C., AND BAKKER, K. 1976. Functional responses in invertebrates. Netherlands J. Zool. 26: 567 572. VAN LENTEREN, J. C., BABENDREIER, D., BIGLER, F., BURGIO, G., HOKKANEN, H. M. T., KUSKE, S., LOOMANS, A. J. M., ME NZLERHOKKANEN, I., VAN RIJN, P. C. J., THOMAS, M. B., TOMMASINI, M. G., AND ZENG, Q. Q. 2003. Environmental risk assessment of exotic natural enemies used in inundative biological control. Biocontrol 48: 3 38. VAN LENTEREN, J. C., BALE, J., BIGLER, F., HOKKANEN, H. M. T., AND LOOMANS, A. J. M. 2006. Assessing risks of releasing exotic biological control agents of arthropod pests Ann. Rev. Entomol. 51: 609 634. VELA, L., DELORENZO, C., AND PEREZ, R. A. 2007. Antioxidant capacity of spanish honeys and its correlation with polyphenol content and other physiochemical properties. J. Sci. Food Agr. 87: 1069 1075. VOEGELE, J., PIZZOL, J., AND BABI, A. 1988. The overwintering of some Trichogramma species. Les Colloques de l INRA 43: 275 282.
246 WACKERS, F. L. 2005. Suitability of (extra) floral nectar, pollen and honeydew as insect food sources, pp. 17 75 In F.L. Wackers, P.C. van Rijn, and J. Bruin [eds.], Plant Provided Food for Carnivarous Insects: A Protective Mutualism and its Applications. Cambr idge University Press, Cambridge, UK. WAAGE, J. K. 2001. Indirect ecological effects in biological control: The challenge and the opportunity, pp. 1 11 In Evaluating Indirect Ecological Effects of Biological Control CABI Publishing, Wallingford, UK. W ALDE, S. J., AND MURDOCH, W. W. 1988. Spatial density dependence in parasitoids. Ann. Rev. Entomol. 33: 441 446. WALLS, J., PIDGEON, N., WEYMAN, A., AND HORLICK JONES, T. 2004. Critical trust: Understanding lay perceptions of health and safety risk regulation. Health Risk Soc 6: 133 150. WANDERMAN, A. 1981. A framework of participation community organizations. J. Appl Behav Sci 17 : 27 58. WA PS HERE, A. J. 1974. A strategy for evaluating the safety of organisms for biological weed control. Ann. Appl. Biol. 77: 201 211. WAPSHERE, A. J. 1989. A testing sequence for reducing rejection of potential biological control agents for weeds. Ann. Appl. Biol. 114: 515526. WARDMAN, J. K. 2008. The constitution of risk communication in advanced liberal societies. Risk Anal. 28: 1619 1637. WARNER, D. K, AND GETZ, C. 2008. A socio economic analysis of the North American commercial natural enemy industry and implications for augmentative biological control. Biol. Control 45: 1 10. WIEDENMANN, R. N., AND S MITH JR, J. W. 1993. Spatial density dependence in parasitoids. Ann. Rev. Entomol. 33: 441 466. W ILCOVE D. S., R OTHSTEIN, D., B UBOW, J., P HILLIPS, A., AND L OSOS E. 1998. Quantifying threats to imperiled species in the United States. Bioscience 48, 607 615. WILCOX, D. 2003. The Guide to Effective Participation [Online] www.partnerships.org.uk/guide. Last accessed 04/05/2011. WILLIAMSON, S. H. 2009. Six Ways to Compute the Relative Value of a U.S. Dollar Amount, 1970 to Present. [Online] http://www.measuringworth.com/uscompare/ Last accessed 04/05/2011. WILLIAMSON, G., AND CARUGHI, A. 2010. Polyphenol content and health benefits of raisins. Nutr. Res. 30: 511 519.
247 WILSON, J. A. 1941. Biological control of Diatraea saccharalis in the Florida everglades during 1940 and 1941. Florida Enomol. 24: 52 57. WILSON J. A. JR, AND D URANT J. A. 1991, Parasites of the European corn borer (Lepidoptera; Pyralidae) in South Carolina, USA. J. Agr Entomol 8 : 109 116 WUHRER, B. G., AND HASSAN, S. A. 1993. Selection of effective species/strains of Trichogramma (Hym., Trichogrammatidae) to control the diamondback moth Plutella xylostella L. (Lep., Plutellidae). J. Appl. Entomol. 116: 80 89. WYNNE, B. 1992. Risk and social learning: Reification to engagement, pp. 275 297 In S. Krimsky D. Golding [eds.], Social Theories of Risk Praeger, Westport, CT. YONG, T H., AND HOFFMANN, M. 2006. H abitat selection by introduced biological control agent Trichogramma ostriniae (Hymenoptera: Trichogrammatidae) and implications for nontarget effects. Behavior 35: 725 732. ZIMMERMANN, H. G., MORAN, V. C., AND HOFFMANN, J. H. 2000. The renowed cactus m oth, Cactoblastis cactorum : Its natural history and threat to native Opuntia floras in Mexico and the United States of America. Diversity Distrib. 6: 259 269. ZIMMERMANN, H. G., MORAN, V. C., AND HOFFMANN, J. H. 2001. The renowned cactus moth, Cactoblastis cactorum (Lepidoptera: Pyralidae): Its natural history and threat to native Opuntia floras in Mexico and the United States of America. Florida Entomol. 84: 543 551. ZIMMERMANN, H. G., AND PREZ SANDI, C. M. 2006. Biology and life cycle of t he cactus moth, pp. 9 17 In H.G. Zimmermann and C.M. Perez Sandi [eds.], The Consequences of Introducing the Cactus Moth ( Cactoblastis cactorum ) to the Caribbean, and Beyond. Transcontinental Reproducciones, Azcapotzalco, Mexico. ZUCCHI, R. A, QUERINO, R. B., AND MONTEIRO, R. C. 2010. Diversity and hosts of Trichogramma in the New World with emphasis in South America. Biol. Control 9: 219 236.
248 BIOGRAPHICAL SKETCH Oulimathe was born in Geneva (Switzerland). She graduated with a Bachelor in micro biology from Laval University, Quebec, (Canada) in 2002. In 2003, she was accepted in the masters program in agricultural sciences at Florida A&M University, Tallahassee (FL). Her work focused on the i solation of t oxic compounds from Xylella fastidiosa, the causative agent of Pierces Disease in g rapev ines Following graduation in 2005, she worked for a year under her Optional Practicum Training (OPT) with the U.S. Department of AgricultureAgricultural Research ServiceCenter for Medical, Agricultural and Veterinary Entomology in Tallahassee (FL). During this year she participated in the control management effort of Cactoblastis cactorum (Berg), a pest of Opuntia spp. in the U.S. and Mexico. In 2006, she started the cooperative Ph.D. program in entom ology between Florida A&M University and University of Florida. Her interest focused in conceptual disciplines such as risk analysis and risk communication combined with an interest in applied entomology, particularly in the area of Classical and Augmenta tive Biological Control. From 2007 to 2011, she was the Florida A&M University representative on the Entomological Society of America South Eastern Branch Student Affair Committee and on the Florida Entomological Society Committee. She received her Ph.D. from the University of Florida in the spring of 2011.