Some Environmental Factors Influencing Rearing of the Spruce
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S AN ABSTRACT OF THE THESIS OF Gary Boyd Pitman for the M. S. in ENTOMOLOGY (Degree) (Major) Date thesis is presented y Title SOME ENVIRONMENTAL FACTORS INFLUENCING REARING OF THE SPRUCE BUDWORM, Choristoneura fumiferana (Clem.) (LEPIDOPTERA: TORTRICIDAE) UNDER LABORATORY CONDITIONS. Abstract approved , (Major Professor) The purpose of this study was to determine the effects of controlled environmental factors upon the development of the spruce budworm (Choristoneura fumiferana Clem.) and to utilize the information for im- proving mass rearing procedures. A standard and a green form of the bud - worm occurring in the Pacific Northwest were compared morphologically and as to their suitability for mass rearing. " An exploratory study demonstrated that both forms of the budworm could be reared in quantity in the laboratory under conditions outlined by Stehr, but that greater survival and efficiency of production would be needed for mass rearing purposes. Further experimentation revealed that, by manipulating environmental factors during the rearing process, the number of budworm generations could be increased from one that occurs normally to nearly three per year. For the standard form of the budworm, procedures were developed for in- creasing laboratory stock twelvefold per generation. Productivity of the green form was much less, indicating that the standard form may be better suited for laboratory rearing in quantity. Recommended rearing procedures consist of the following steps. Egg masses should be incubated at temperatures between 70 and 75 °F and a relative humidity near 77 percent. Under these conditions, embryo matur- ation and hibernacula site selection require approximately 8 to 9 days. The larvae should be left at incubation conditions for no longer than three weeks. A pre- conditioning temperature treatment, one week at 45°F, appears necessary before larvae in hibernacula are exposed to overwinter- ing conditions. The cold rest requirement is fulfilled partially by holding the larvae for 52 days at a constant temperature of 33 °F. This period appears to be optimal in terms of survival and minimum storage time. Completion of the resting period is brought about by constant ± illumination on a fluorescent light table at a temperature of 75 3 °F. Approximately 26 days is required for total larval issuance. 2 Newly issued larvae should be collected daily and confined in half - pint ice cream cartons containing newly prepared or freshly thawed, agar - coated Douglas -fir seedlings. The optimum number of larvae per carton has not been determined; however 20 per feeding container appears to be the maximum for uninhibited development. The first food change is governed by the rate of deterioration of food and is usually necessary 10 days after larval introduction. Two or three more food changes may be required; this will depend on the number of surviving larvae. When held at nearly 75°F, most larvae mature and pupate in approximately 30 days. Adult eclosion occurs within 7 to 8 days; absorbent substrate was found useful in aiding eclosion. Satisfactory mass mating and oviposition can be obtained by using a large, walk -in cage containing young potted Douglas -fir trees. Egg masses can be concentrated on one or two trees at one corner of the cage by use of low intensity light at this corner. Studies of photomicrographs of sectioned thoracic cuticles of 1 -day- old pupae and empty pupae cases.. revealed groins dissimilarities between the two forms of the budworm. The outer region of the standard form's cuticle was markedly darker than the green form which suggests a differ- ence in exocuticle composition. However, no significant variation in total cuticle thickness or exocuticle thickness could be determined. No dif- ferences were found in the cuticle of mature larvae of the two forms. Characteristics of the cuticle were noted in detail. The ventral portion of the thoracic cuticle is discontinuous. Each appendage is separated from internal structures by a membrane appearing to be an ex- tension of the outer cuticle. Integument rigidity is maintained through a bridle structure which is fastened to the outer cuticle of each forming appendage. The type of bridle associated with the forming wings was similar in both forms. The various bridle structures varied in size be- tween the two forms. -+ SOME ENVIRONMENTAL FACTORS INFLUENCING REARING OF THE SPRUCE BUDWORM, CHORISTONEURA FUMIFERANA (CLEM.) (LEPIDOPTERA: TORTRICIDAE) UNDER LABORATORY CONDITIONS by GARY BOYD PITMAN A THESIS submitted to OREGON STATE UNIVERSITY in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE June 1963 APPROVED: Associate Professor of Entomology Chairman of Department of Entomology Chairman of School raduate ommittee Dean of Graduate School Date thesis is presented 2,z /, /G 2- Typed by Helen Sewell ACKNOWLEDGMENTS Acknowledgment should first be made to the Pacific Northwest Forest and Range Experiment Station, U. S. Forest Service, and the Department of Entomology, Oregon State University. The cooperative agreement between the two organizations was the basis for this study. The author is grateful to several people whose guidance and assitance made the completion of this study possible. Specific acknowledgments are due, as follows: To Dr. Julius A. Rudinsky, Professor of Entomology, Oregon State University, for advice and criticism on all phases of this study. To Dr. Paul O. Ritcher, Chairman, Department of Entomology, Oregon State University, for assistance in planning this research. To Mr. Valentine M. Carolin Jr., Entomologist, Pacific Northwest Forest and Range Experiment Station, for his constant guidance and encouragement throughout the entire period of study. To Mr. Robert L. Furniss, Chief, Division of Forest Insect Research, Pacific Northwest Forest and Range Experiment Station, for constructive criticism of work plans and for reviewing the manuscript. To Dr. Clarence G. Thompson, Insect Pathologist, Pacific Northwest Forest and Range Experiment Station, for identification of pathogenic microorganisms encountered during the rearing program. To Dr. Roger B. Ryan, Entomologist, Pacific Northwest Forest and Range Experiment Station, for drawing Figure 5. To Mr. Wallace C. Guy, Pacific Northwest Forest and Range Experiment Station, for photographic assistance. To Mr. Thomas Onsdorff, Associate Professor of Food Technology, Oregon State University, for making available the walk -in coolers used in this research. TABLE OF CONTENTS Page INTRODUCTION 1 Status as a Forest Pest t , 1 Justification for Study 2 3 . Objectives. ., . , .6 REVIEW OF LITERATURE . r 4 Life History . -, k. e 4 Diapause - 6 Rearing Techniques e ,. .. .. 11 MATERIALS AND METHODS w Y 16 Obtaining Experimental Stock . 16 Laboratory Rearing Conditions 17 Mating and Oviposition . , - 18 Walk -in cage 18 Gallon jars r 4 . .. ., 18 Plastic boxes _ . 19 Incubation and Pre -diapause .. 21 Petri dishes - - 21 Ice cream cartons . 21 Light table , , . .. 23 Growth chambers 23 Diapause Is . , 24 Walk -in coolers i 24 Growth chamber modified for variable temperatures . 24 Post -diapause . 27 Handling procedure . , 4 , 27 Natural food . 28 Modified food _ t - 29 Feeding procedure - 30 Preparation of Cuticle Sections . .. 32 Larvae and pupae ti M 32 Pupal case - 35 TABLE OF CONTENTS - Continued Page RESULTS 36 Preliminary Study 36 Environmental Studies 39 Fecundity and incubation , 39 Hibernacula selection 53 Incidence of non -diapause 54 Pre -diapause treatments ...... .. , . 55 Effects of a fluctuating . cold storage temperature . 64 Role of temperature and light in terminating diapause . 66 Effect of a thermal shock treatment during diapause development 72 Modified food ,. .. ,r 75 Comparative Morphological Studies 4, 83 DISCUSSION - . ., 93 98 CONCLUSIONS . - - LIST OF FIGURES Figure Page 1. Large walk -in cage used for mass mating and oviposition , 20 2. One gallon jar mating confinements 20 3. Stainless steel tray and individual plastic boxes used for individual mating confinements 22 4. Modified half -pint ice cream carton with egg -bearing needles cemented to lid 22 5. Diagrammatic view of the essential parts of the temperature programming device 26 6. Wooden rack used to hold ice -cream cartons during feeding procedure , . 31 7. Relation of percent egg survival to relative humidity at three chamber temperatures as compared with standard rearing conditions 52 8. Effects of various pre -diapause treatments on larval issuance following 6, 8, 10, and 12 weeks cold rest (Green form) - - 62 9. Summary of larval survival during diapause development after 4 different periods of cold rest, as determined by the mean percent survival for various pre -diapause treatment groups. (Green form) . .. , . 63 10. Effect of fluorescent and incandescent illumination on diapause termination in larvae receiving different periods of cold rest. (Standard form) . 71 11. Photomicrograph of a 10 micron X- section (79.3X) through the posterior margin of the prothorax of a 1 -day old pupa (Standard form) ' 85 12. Photomicrograph of a 10 micron X- section (16X) through the posterior margin of the prothorax of a 1 -day pupa - old (Standard form) , 85 13. Photomicrograph of a 10 micron X- section (79.3X) through the posterior margin of the prothorax of a 1 -day old pupa (Green form) 86 LIST OF FIGURES - Continued Page Figure 14. Photomicrograph of a 10 micron X- section (16X) through the posterior margin of the prothorax of a 1 -day old pupa (Green form) . 86 15. Photomicrographs of a 10 micron