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Diflubenzuron: control of the codling moth, Laspeyresia pomonella L., and the obliquebanded leafroller,… Anderson, Dale William 1980

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DIFLUBENZURON:  CONTROL OF THE CODLING MOTH, Laspeyres  pomonella L.,  AND THE OBLIQUEBANDED LEAFROLLER,  C h o r i s t o n e u r a rosaceana  (Harris)  by lALE WILLIAM ANDERSON B.Sc.  ( A g r . ) , U n i v e r s i t y o f B r i t i s h C o l u m b i a , 1978  A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE  in THE FACULTY OF GRADUATE STUDIES (Department o f P l a n t  Science)  We a c c e p t t h i s t h e s i s as conforming t o the r e q u i r e d s t a n d a r d  THE UNIVERSITY OF BRITISH COLUMBIA December, 1980 @  Dale W i l l i a m A n d e r s o n , 1980  In p r e s e n t i n g t h i s  thesis  an advanced degree at  further  agree  fulfilment  of  the  requirements  the U n i v e r s i t y of B r i t i s h Columbia, I agree  the L i b r a r y s h a l l make it I  in p a r t i a l  freely  available  for  this  thesis  f o r s c h o l a r l y purposes may be granted by the Head of my Department  of  this  thesis for  It  financial  of  gain s h a l l not  PLANT SCIENCE  The U n i v e r s i t y o f B r i t i s h Columbia 2075 Wesbrook P l a c e V a n c o u v e r , Canada V6T 1W5  DECEMBER 16, 1980  or  i s understood that copying or p u b l i c a t i o n  written permission.  Department  that  reference and study.  t h a t p e r m i s s i o n f o r e x t e n s i v e copying o f  by h i s r e p r e s e n t a t i v e s .  for  be allowed without my  -ii -  ABSTRACT Diflubenzuron  ( D i m i l i n ) was e v a l u a t e d as a c o n t r o l  agent f o r the c o d l i n g  moth, L a s p e y r e s i a pomonella L. and obiiquebanded l e a f r o l l e r , rosaceana  Choristoneura  (Harris).  L a b o r a t o r y t e s t s showed c o d l i n g moth eggs to be e x t r e m e l y s e n s i t i v e d i f l u b e n z u r o n , p a r t i c u l a r l y when t r e a t e d t o p i c a l l y tion.  LD50  values f o r 0- to  respectively.  immediately a f t e r  to  oviposi-  and 3 - d a y - o l d eggs were 1.1 and 17.2 ppm,  D i f l u b e n z u r o n was more e f f e c t i v e as an o v i c i d e when kept  s o l u t i o n on egg s u r f a c e s f o r l o n g e r t i m e p e r i o d s .  in  When the compound had  d r i e d on f r u i t or f o l i a g e , r e s i d u a l a c t i o n was e x c e l l e n t a g a i n s t t h e eggs. T h i s a c t i v i t y d i d not decrease over a 10-day p e r i o d and d a i l y s p r a y i n g o f f o l i a g e w i t h water d i d not s i g n i f i c a n t l y reduce o v i c i d a l a surfactant,  action.  Tween 20,  improved the d i s t r i b u t i o n and e f f i c a c y o f d i f l u b e n z u r o n on  f o l i a g e and young a p p l e s but not waxy mature a p p l e s . F i r s t - and 2 n d - i n s t a r c o d l i n g moth l a r v a e were s u s c e p t i b l e to d i f l u b e n zuron when t h e compound was i n g e s t e d from agar d i e t o r a p p l e f o l i a g e .  On t h e  d i e t , the L D g ' s f o r the two i n s t a r s were 48.2 and 8.1 ppm, r e s p e c t i v e l y . 5  A d u l t c o d l i n g moths dipped i n or f e d d i f l u b e n z u r o n s o l u t i o n s adverse e f f e c t s , a l t h o u g h marginal  showed no marked  r e d u c t i o n s i n f e c u n d i t y or egg v i a b i l i t y  were o b s e r v e d . The s e n s i t i v i t y o f the obiiquebanded l e a f r o l l e r  (a r e p r e s e n t a t i v e  r o l l e r s p e c i e s ) was a l s o examined under l a b o r a t o r y c o n d i t i o n s .  leaf-  Egg masses  were i n s e n s i t i v e t o d i f l u b e n z u r o n a t c o n c e n t r a t i o n s as h i g h as 1,000 ppm. However, 100 ppm d i f l u b e n z u r o n proved t o x i c t o f i r s t - i n s t a r duced l o n g e v i t y o f a d u l t moths.  l a r v a e and r e -  In f i e l d t e s t s , d i f l u b e n z u r o n s i g n i f i c a n t l y reduced l e a f r o l l e r to apples.  damage  However, a t comparable a p p l i c a t i o n r a t e s (375 ppm), c o n t r o l was  o n l y 25% t h a t p r o v i d e d by a z i n p h o s - m e t h y l  (Guthion).  the c o d l i n g moth d i f l u b e n z u r o n p r o v i d e d c o n t r o l  In c o n t r a s t ,  against  equal to t h a t o f a z i n p h o s -  methyl i n o r c h a r d s o f Golden D e l i c i o u s and mixed a p p l e c u l t i v a r s . a d d i t i o n o f Tween 20 d i d not enhance the e f f i c a c y o f d i f l u b e n z u r o n .  The Leaf  m i t e - c o u n t s suggested t h a t d i f l u b e n z u r o n was n o n - t o x i c to predaceous m i t e s . The i m p l i c a t i o n s o f t h e s e f i n d i n g s a r e d i s c u s s e d i n terms o f t h e p o t e n tial  of diflubenzuron in the i n t e g r a t e d c o n t r o l of apple p e s t s .  - i v-  TABLE OF CONTENTS  Raje ABSTRACT  i i  LIST OF TABLES  v  LIST OF FIGURES  vii  LIST OF PLATES  viii  ACKNOWLEDGEMENTS  ix  INTRODUCTION  1  MATERIALS AND METHODS  12  A.  R e a r i n g Techniques  12  B.  T e s t Compounds  13  C.  T e s t s on Eggs  16  D.  T e s t s on Larvae  17  E.  T e s t s on A d u l t s  17  F.  F i e l d Tests  19  G.  Statistical  Analysis  RESULTS I.  23 24  Laboratory Studies  24  A.  C o d l i n g Moth  24  B.  Obiiquebanded L e a f r o l l e r  37  I I . F i e l d Tests  42  A.  L e a f r o l l e r Test  42  B.  C o d l i n g Moth T e s t s  48  DISCUSSION  54  LITERATURE CITED  61  APPENDIX  68  7  -V-  LIST OF TABLES Table  Page  I  P e r c e n t hatch and c o n t r o l o f c o d l i n g moth eggs t r e a t e d t o p i c a l l y w i t h v a r y i n g c o n c e n t r a t i o n s of d i f l u b e n z u r o n .  25  II<  Maximum l i k e l i h o o d t e s t s o n , t h e t o x i c e f f e c t s o f d i f l u benzuron to eggs o f the c o d l i n g moth.  26  III  P e r c e n t hatch o f c o d l i n g moth eggs t r e a t e d t o p i c a l l y w i t h 10 ppm d i f l u b e n z u r o n a t v a r y i n g times from o v i position.  27  IV  Perpent hatch o f c o d l i n g moth eggs p l a c e d i n c o n t a c t w i t h 10 ppm d i f l u b e n z u r o n s o l u t i o n s f o r v a r y i n g p e r i o d s of time.  ,. '  29 .  1  V  P e r c e n t hatch o f c o d l i n g moth eggs t r e a t e d w i t h d i f l u benzuron s o l u t i o n s i n the presence and absence o f Tween 20.  30  VI  . . . H a t c h a b i l i t y of c o d l i n g 'moth eggs o v i p o s i t e d on a p p l e s which had been d i p p e d - i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s .  31  VII  H a t c h a b i l i t y o f c o d l i n g moth eggs o v i p o s i t e d on pear < . f o l i a g e which had been dipped in' d i f l ubenzuron/Tween 20 solutions  33  VI IT  M o r t a l i t y o f f i r s t - and 2 n d - i n s t a r c o d l i n g moth l a r v a e . f e e d i n g on agar medium c o n t a i n i n g v a r i o u s c o n c e n t r a t i o n s of d i f l u b e n z u r o n .  34  IX  Maximum l i k e l i h o o d t e s t s on the t o x i c e f f e c t s o f d i f l u benzuron to l a r v a e o f the c o d l i n g moth 7  35  X  P e n e t r a t i o n o f a p p l e s and s u r v i v a l t o 4 t h - i n s t a r by c o d l i n g moth f i r s t - i n s t a r l a r v a e , a f t e r f e e d i n g on a p p l e f o l i a g e sprayed w i t h d i f l u b e n z u r o n / T w e e n 20.  36  XI  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t c o d l i n g moths dipped i n d i f l u b e n z u r o n s o l u t i o n s .  38  XII  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t c o d l i n g moths dipped i n d i f l ubenzuron/Tween 20 s o l u t i o n s .  39  XIII  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y of a d u l t c o d l i n g moths f e d d i f l u b e n z u r o n s o l u t i o n s . "  40  'i  •  . .  .  -VI-  Table  Page  XIV  P e r c e n t hatch o f l e a f r o l l e r egg masses t o p i c a l l y t r e a t e d with varying concentrations of d i f l u b e n z u r o n .  41  XV  Percent s u r v i v a l of f i r s t - i n s t a r l e a f r o l l e r larvae f e e d i n g on pear f o l i a g e which had been dipped i n dif1ubenzuron/Tween 20 s o l u t i o n s .  43  XVI  M o u l t i n g success o f f i v e l e a f r o l l e r l a r v a l i n s t a r s p l a c e d on a p p l e f o l i a g e sprayed w i t h d i f l u b e n z u r o n / Tween 20 s o l u t i o n s .  44  XVII  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y of a d u l t l e a f r o l l e r moths dipped i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s .  45  XVIII  P e r c e n t o f h a r v e s t e d a p p l e s showing l e a f r o l l e r damage.  47  XIX  P e r c e n t o f h a r v e s t e d a p p l e s o f mixed c u l t i v a r s showing c o d l i n g moth damage.  50  XX  Percent o f h a r v e s t e d Golden D e l i c i o u s a p p l e s showing c o d l i n g moth damage.  52  LIST OF FIGURES Figure 1  Schematic r e p r e s e n t a t i o n o f t h e c o d l i n g moth f i e l d test orchards.  2  Schematic r e p r e s e n t a t i o n o f the l e a f r o l l e r orchard.  3  Seasonal abundance o f f o u r l e a f r o l l e r s p e c i e s i n t h e t e s t o r c h a r d d u r i n g summer, 1980.  4  Seasonal abundance o f the c o d l i n g moth i n Summerland o r c h a r d s d u r i n g summer, 1980.  5  European red m i t e numbers, per l e a f and per p h y t o s e i i d , i n the c o d l i n g moth f i e l d - t e s t o r c h a r d s d u r i n g summer, 1980.  field-test  LIST OF PLATES Plate I  C o r r u g a t e d wooden s t r i p s used f o r c o l l e c t i o n c o d l i n g moth l a r v a e .  of  II  R o t a t i n g o v i p o s i t i o n cage used f o r c o l l e c t i o n c o d l i n g moth eggs.  of  III  Broad beans, used f o r r e a r i n g o f l e a f r o l l e r s , i n a greenhouse s i t u a t i o n .  growing  IV  Wax paper cage used f o r experiments on a d u l t moths and f o r c o l l e c t i o n o f l e a f r o l l e r eggs.  V  C l i p - c a g e used f o r t e s t s on f i r s t - i n s t a r c o d l i n g moth larvae  VI  A p p l e f o l i a g e showing f e e d i n g i n j u r y by c o d l i n g moth f i r s t - i n s t a r larvae.  -ix-  ACKNOWLEDGEMENTS The a u t h o r wishes to e x p r e s s h i s s i n c e r e a p p r e c i a t i o n to the members o f h i s committee:  Dr. G.W. E a t o n , Dr. R.H. E l l i o t t ,  Dr. R.D. McMullen and Dr. V . C . R u n e c k l e s .  A special  Dr. D.G.  Finlayson,  thanks must go to  Dr. McMullen and the o t h e r entomology personnel a t the Summerland Research S t a t i o n , f o r i n v a l u a b l e a s s i s t a n c e w i t h the f i e l d t e s t i n g . grateful with this  to my s u p e r v i s o r , Dr. R.H. E l l i o t t ,  I am p a r t i c u l a r l y  f o r h i s a d v i c e and a s s i s t a n c e  project.  I would a l s o l i k e t o thank A g r i c u l t u r e Canada and Dr. E l l i o t t p r o v i d i n g my s u p p o r t .  for  -1-  INTRODUCTION The c o d l i n g moth, L a s p e y r e s i a pomonella L.  (Lepidoptera:  O l e t h r e u t i d a e ) , i s g e n e r a l l y regarded as the s i n g l e most d e s t r u c t i v e o f pome f r u i t s  i n the w o r l d .  Since i t s  i n t r o d u c t i o n to North America  New England c a . 1750, the c o d l i n g moth has spread t o v i r t u a l l y a l l where pome f r u i t s  pest  a r e grown ( S l i n g e r l a n d and Crosby 1915).  regions  A p p l e s and  pears are the main economic hosts but the l a r v a e have o c c a s i o n a l l y found i n peaches, a p r i c o t s , c h e r r i e s and v a r i o u s n u t s .  via  Infested  been  fruit  g e n e r a l l y show the c h a r a c t e r i s t i c symptom o f e n t r y holes clogged w i t h frass.  In u n p r o t e c t e d o r c h a r d s , e s p e c i a l l y i n commercial  p r o d u c t i o n areas  and warmer c l i m a t e s , l o s s e s may r e a c h 95% o r g r e a t e r ( M e t c a l f e t aJL Virtually all  1962).  t r e e - f r u i t p r o d u c t i o n i n B r i t i s h Columbia o c c u r s i n the  Okanagan and Similkameen V a l l e y s , w i t h a s m a l l e r i n d u s t r y i n the E a s t Kootenay.  The c o d l i n g moth t y p i c a l l y undergoes two or t h r e e  per y e a r i n southern B r i t i s h C o l u m b i a .  generations  The mature l a r v a e o v e r w i n t e r  within  t h i n s i l k e n cases underneath l o o s e bark or i n the s o i l and pupate i n the spring. apple.  A d u l t moths begin t o emerge as e a r l y as the blossom p e r i o d o f The peak o f a d u l t emergence o c c u r s t h r e e t o f o u r weeks l a t e r depen-  d i n g on weather c o n d i t i o n s .  Mating a c t i v i t y o f moths i s reduced a t below  15°C, but p r o v i d e d t h a t warm temperatures p r e v a i l , females begin t o o v i p o s i t on f r u i t o r f o l i a g e t h r e e to f i v e days a f t e r emergence.  The newly-  hatched l a r v a e may f e e d f o r a s h o r t time on f o l i a g e b e f o r e e n t e r i n g the f r u i t , o f t e n a t the c a l y x end.  The l a r v a e mature i n a p p r o x i m a t e l y  three  weeks and l e a v e the f r u i t t o pupate on t w i g s , b r a n c h e s , under bark or the s o i l .  in  Peak emergence o f a d u l t s o f the second brood o c c u r s d u r i n g m i d -  -2-  J u l y through mid-August depending on weather c o n d i t i o n s .  In y e a r s w i t h  v e r y warm summers, a t h i r d f l i g h t o f moths may o c c u r i n September,  just  b e f o r e normal a p p l e h a r v e s t i n g dates (Madsen and A r r a n d 1971). As even r e l a t i v e l y low p o p u l a t i o n s o f the c o d l i n g moth can cause s e r i o u s damage, c o n t r o l measures a r e v i t a l . by a r e s i d u a l  The f r u i t must be p r o t e c t e d  s p r a y a t a time when the a d u l t moths a r e p r e s e n t .  In p a s t  y e a r s , compounds such as l e a d a r s e n a t e and DDT p r o v i d e d good c o n t r o l , the development o f r e s i s t a n c e  (Marshall  but  1959; Putman 1962) a l o n g w i t h un-  d e s i r a b l e e f f e c t s on the environment (Newsom 1967) c u r t a i l e d t h e i r  use.  Three organophosphate i n s e c t i c i d e s a r e p r e s e n t l y recommended f o r c o d l i n g moth c o n t r o l .  Azinphos-methyl  (Guthion) i s most w i d e l y used and  i s n o r m a l l y a p p l i e d s h o r t l y a f t e r the appearance o f a d u l t moths.  T h i s com-  pound i s a broad-spectrum i n s e c t i c i d e which does not c o n c e n t r a t e i n food c h a i n s and p r o v i d e s e x c e l l e n t c o n t r o l  i f used c o r r e c t l y .  Its  residual  a c t i v i t y may be as l o n g as two weeks on f o l i a g e , p r o v i d i n g extended c o n t r o l ( M a r t i n 1971).  Furthermore, i t s t o x i c i t y to predatory mites i s  allowing i n t e g r a t i o n with b i o l o g i c a l mite control At the same t i m e , a z i n p h o s - m e t h y l oral  low,  (Downing and A r r a n d 1978).  i s a v e r y t o x i c p e s t i c i d e , w i t h an acute  LDgg ( r a t s ) o f 16.4 mg/kg ( M a r t i n 1971).  The use o f t h i s  insecticide  p r e s e n t s a hazard to t h e a p p l i c a t o r , who must wear a r e s p i r a t o r and p r o t e c tive clothing.  As the high t o x i c i t y and long p e r s i s t e n c e o f  azinphos-  methyl p r e v e n t i t s use w i t h i n seven days o f h a r v e s t , t h e compound i s not recommended f o r c o n t r o l o f a t h i r d c o d l i n g moth brood (Anonymous 1980). Phosalone ( Z o l o n e ) and phosmet (Imidan) a r e a l s o broad-spectrum  insec-  t i c i d e s not b i o l o g i c a l l y c o n c e n t r a t e d ( M a r t i n 1971) and o f low t o x i c i t y p r e d a t o r y m i t e s (Downing and A r r a n d 1978).  to  Both a r e o f lower t o x i c i t y and  -3-  p e r s i s t e n c e than a z i n p h o s - m e t h y l  and can be a p p l i e d up to one day b e f o r e  h a r v e s t on a p p l e s s o l d i n Canada (Anonymous 1980).  C o n t r o l e f f e c t e d by  t h e s e two compounds does not u s u a l l y r e a c h t h a t o f a z i n p h o s - m e t h y l . t h r e e organophpsphates have been observed t o d i s r u p t n o n - t a r g e t including natural  enemies o f the pear p s y l l a , P s y l l a p y r i c o l a  (McMullen and Jong 1967; W e s t i g a r d 1979). moth c o n t r o l a r e h i g h l y  All  species,  Foerster  A l t e r n a t i v e methods o f  codling  desirable.  The a v a i l a b i l i t y o f the s y n t h e t i c c o d l i n g moth sex pheromone p r o v i d e s several  possible control  techniques.  The " c o n f u s i o n " t e c h n i q u e  r e l e a s e o f the pheromone over a g i v e n a r e a i n amounts s u f f i c i e n t males from l o c a t i n g f e m a l e s . t h i s method.  Moffitt  involves to prevent  (1978) demonstrated good p o t e n t i a l  A second t e c h n i q u e i n v o l v e s m a s s - t r a p p i n g and removal  males u s i n g sex pheromone t r a p s .  for  of  Hagley (1978) suggested t h a t t h i s was  q u i t e f e a s i b l e , but Madsen and C a r t y (1979) showed t h a t the t e c h n i q u e was e f f e c t i v e o n l y i f c o d l i n g moth p o p u l a t i o n s were low and i f cluded r e i n f e s t a t i o n .  isolation  In any c a s e , the sex pheromone a l l o w s  pre-  accurate  m o n i t o r i n g o f pest l e v e l s and more p r e c i s e a p p l i c a t i o n o f chemical  controls  (Myburgh et al_. 1974). The use o f b i o l o g i c a l considerable attention.  agents a g a i n s t the c o d l i n g moth has r e c e i v e d  Cox (1932) r e a r e d l a r g e numbers of A s c o g a s t e r  carpocapsae V i e r e c k (Hymenoptera: moth l a r v a e i n New Y o r k .  B r a c o n i d a e ) from f i e l d - c o l l e c t e d  Russ and B r a n d s t a t t e r (1978) r e p o r t e d  progress  i n the r e a r i n g o f A s c o g a s t e r q u a d r i d e n t a t u s Wesm., another i m p o r t a n t o f t h e c o d l i n g moth.  parasite  Voegele e_t aJL (1978) showed a r e d u c t i o n i n c o d l i n g  moth damage a f t e r r e l e a s e o f the egg p a r a s i t e s Trichogramma s p p . tera:  codling  Trichogrammatidae).  In another form "of b i o l o g i c a l  (Hymenop-  c o n t r o l , Solomon  -4-  e t aj_. (1976) showed t h a t b i r d s i n c e r t a i n E n g l i s h o r c h a r d s took up t o 95% o f t h e o v e r w i n t e r i n g c o d l i n g moth l a r v a e .  Microbial  agents such as  B a c i l l u s t h u r i n g i e n s i s and the entomopathogenic fungus B e a u v e r i a  bassiana  have shown l i m i t e d s u c c e s s ( F e r r o n and V i n c e n t 1978).  control  Excellent  has been o b t a i n e d u s i n g g r a n u l o s i s v i r u s a g a i n s t the c o d l i n g moth  (Fischer-  Col br>ie 1978; D i c k l e r and Huber 1978) even though t h i s method has been shown to be somewhat i n c o n s i s t e n t two most p r o m i s i n g non-chemical virus(es)  (Huber and D i c k l e r 1978).  P r o b a b l y the  c o n t r o l methods are t h e use o f  granulosis  and the r e l e a s e o f s t e r i l i z e d a d u l t c o d l i n g moths.  An i n t e n s i v e e v a l u a t i o n o f the s t e r i l e  i n s e c t t e c h n i q u e as a c o n t r o l  measure a g a i n s t the c o d l i n g moth was i n i t i a t e d i n 1962 as a p r o j e c t o f the A g r i c u l t u r e Canada Research S t a t i o n a t Summerland, B.C. expanded u n t i l  i n 1976, 1977 and 1978 i t  i n c l u d e d the g r e a t e r p a r t o f  a p p l e and pear o r c h a r d s i n the Similkameen V a l l e y . encouraging.  T h i s program all  The r e s u l t s were most  D e s p i t e high c o s t s , the economic f e a s i b i l i t y o f the program  was i n c r e a s e d by improved e f f i c i e n c y  i n r e a r i n g and r e l e a s e  techniques  ( P r o v e r b s e t aj_. 1977). The a d u l t moths are r e a r e d on a s y n t h e t i c d i e t a t the Summerland Research S t a t i o n and s t e r i l i z e d w i t h 35 krad o f gamma r a d i a t i o n t e l y p r i o r to r e l e a s e .  immedia-  For optimum c o n t r o l , a r a t i o o f 40 s t e r i l e males  t o one n a t i v e male i s d e s i r a b l e ( P r o v e r b s 1978).  To ensure t h a t t h i s  ratio  i s m a i n t a i n e d , p o p u l a t i o n l e v e l s o f n a t i v e and r e l e a s e d s t e r i l e moths a r e c o n s t a n t l y monitored w i t h sex pheromone t r a p s . t i f i e d by the presence i n t e r n a l l y o f c a l c o o i l  S t e r i l e moths can be i d e n r e d , a dye p l a c e d i n the  l a r v a l d i e t ( B r i n t o n e t aj_. 1969). As the s t e r i l e  i n s e c t t e c h n i q u e i s g e n e r a l l y f e a s i b l e o n l y when  -5-  a p p l i e d a g a i n s t low p e s t p o p u l a t i o n s , c o n c e r t e d e f f o r t s were made to reduce n a t i v e c o d l i n g moth p o p u l a t i o n s to the r e l e a s e o f s t e r i l e moths.  i n the Similkameen V a l l e y  prior  Measures taken i n c l u d e d removal  abandoned host t r e e s and i n t e n s i v e s p r a y i n g o f badly i n f e s t e d  of  orchards.  During the c o u r s e o f the program, i n s e c t i c i d e s were a p p l i e d to areas which w i l d p o p u l a t i o n s o f c o d l i n g moths r o s e to damaging l e v e l s s t e r i l e i n s e c t r e l e a s e ( P r o v e r b s 1978). insecticides s t i l l  in  despite  I t can be seen, t h e r e f o r e ,  that  have a s i g n i f i c a n t r o l e i n e n s u r i n g the success o.f-..a  s t e r i l e c o d l i n g moth program. Although azinphos-methyl  i s c u s t o m a r i l y used when sprays are deemed  n e c e s s a r y , the compound i s p o t e n t i a l l y h i g h l y t o x i c t o the n o n - t a r g e t organisms and t o a d u l t c o d l i n g moths.  applicator,  Because o f t o x i c i t y  t h e s t e r i l e moths and t h e - p e r s o n idi.spensing them, s t e r i l e  insect  to  release  i s c u r t a i l e d i n sprayed o r c h a r d s f o r t h r e e to seven days (M.D. P r o v e r b s , personal communication).  Release o f s t e r i l e c o d l i n g moths must be resumed  at or before c e s s a t i o n of e f f e c t i v e r e s i d u a l p r o t e c t i o n o f the o r c h a r d i s i n c o m p l e t e . i n s e c t i c i d e w i t h good r e s i d u a l  chemical c o n t r o l ,  otherwise  A l e s s t o x i c , .more , p e s t - s p e c i f i c  a c t i v i t y would be p r e f e r a b l e t o a z i n p h o s -  methyl whether used a l o n e or i n c o n j u n c t i o n w i t h s t e r i l i t y - c o n t r o l .  If  r e l e a s e o f aduTt"moths was c o n t i n u o u s , an i n s e c t i c i d e o f lower t o x i c i t y a d u l t moths but e f f e c t i v e a g a i n s t immature "stages would d i s r u p t c o n t r o l to a l e s s e r  sterility-  extent.  The i n s e c t growth r e g u l a t o r d i f l u b e n z u r o n ( D i m i l i n ) may s a t i s f y criteria.  to  When a p p l i e d a t v e r y - l o w c o n c e n t r a t i o n s  (van Daalen e t a]_.  these 1972),  the compound has e f f e c t i v e l y c o n t r o l l e d many p e s t s p e c i e s w i t h i n the major i n s e c t o r d e r s , p a r t i c u l a r l y D i p t e r a , C o l e o p t e r a and L e p i d o p t e r a .  In the  -6-  l a t t e r o r d e r , t h e gypsy moth P o r t h e t r i a d i s p a r  (L.)  ( G r a n e t t and Dunbar  1 9 7 5 ) , the z e b r a c a t e r p i l l a r Ceramica p i e t a ( H a r r i s )  (Tamaki and Turner  1974), the f o r e s t t e n t c a t e r p i l l a r Malacosoma d i s s t r i a  (Hbn.)  (Retnakaran  e t al_. 1979), the D o u g l a s - f i r t u s s o c k moth O r y g i a pseudotsugata  (McDunnough)  ( N e i s e s s e t al_. 1976) and o t h e r pests have been shown to be s u s c e p t i b l e diflubenzuron. Culicidae)  Research conducted on Culex pi p i ens p i pi ens  suggests t h a t r e s i s t a n c e to the growth r e g u l a t o r  much more s l o w l y than t o c o n v e n t i o n a l  insecticides  to  (Diptera: accumulates  (Brown e t aj_. 1978).  There are r e p o r t s which suggest t h a t the c o d l i n g moth may be s e n s i t i v e  to  the compound (Anderson 1978; Audemard 1978; Cranham 1978; Wearing and Thomad 1978; W e s t i g a r d 1979) and the v e r i f i c a t i o n o f t h i s i s the major aim of t h i s  study.  I t appears t h a t d i f l u b e n z u r o n , a stomach p o i s o n , i n h i b i t s  the  chitin  s y n t h e t a s e enzyme i n v o l v e d i n the s y n t h e s i s and d e p o s i t i o n o f new c u t i c l e ( P o s t and V i n c e n t 1973; Mayer e_t al_. 1980).  E f f e c t s on i n s e c t s a r e  n o t i c e a b l e d u r i n g the moult i n t h a t s u s c e p t i b l e immatures a r e  usually  incapable  o f shedding t h e i r e x u v i a e or form t h i n and deformed c u t i c l e s which r u p t u r e easily.  Ovicidal  qualities  have been observed i n t e s t s on mosquitoes  (Muira e t al_. 1 9 7 6 ) , b l a c k f l i e s  (Lacey and M u l l a 1977), armyworms  (Ascher  and Nemny 1976) and the c o d l i n g moth (Anderson 1978; Wearing and Thomas 1978).  A d u l t c h e m o s t e r i l i z a t i o n has been r e c o r d e d f o r mosquitoes  e t al_. 1976), b o l l w e e v i l s s t a b l e f l i e s and horn f l i e s  (Muira  (Moore and T a f t 1975; Johnson e t aj_. 1978), (Wright and H a r r i s 1976).  C l a r k e e t al_. (1977)  showed f e e d i n g d e t e r r e n t p r o p e r t i e s a g a i n s t L o c u s t a m i g r a t o r i a .  The modes  o f a c t i o n o f d i f l u b e n z u r o n as an o v i c i d e , c h e m o s t e r i l a n t and f e e d i n g d e t e r r e n t a r e not f u l l y  understood.  -7-  D i f l u b e n z u r o n i s g e n e r a l l y f o r m u l a t e d as a w e t t a b l e powder.  This  may remain a c t i v e up to s i x months i n s t o r e d g r a i n (McGregor and Kramer 1 9 7 6 ) , t h r e e weeks on l e a f s u r f a c e s  (Sahota and Shepard 1 9 7 5 ) ,  three  weeks i n water ( M u l l a and Darwazeh 1 9 7 5 ) and has a h a l f - l i f e o f about two months i n d r y s o i l .  D i f l u b e n z u r o n i s n o n - v o l a t i l e and q u i t e s t a b l e  extremes o f temperature and l i g h t . at  25°C  ( M e t c a l f e t al_.  I t s water s o l u b i l i t y  in  i s about 1 . 0 ppm  1975).  Research has been conducted i n t o o t h e r f o r m u l a t i o n s as w e . l l .  Taft  and Hopkins ( 1 9 7 5 ) o b t a i n e d 98% s t e r i l i z a t i o n o f a d u l t b o l l w e e v i l s by a p p l y i n g d i f l u b e n z u r o n i n a s p r a y a b l e b a i t o f i n v e r t sugar and m o l a s s e s . L l o y d e_tal_. ( 1 9 7 7 ) a p p l i e d d i f l u b e n z u r o n i n c o t t o n s e e d o i l weevils, with excellent r e s u l t s .  Other o i l  formulations  against  boll  have been i n v e s -  t i g a t e d and hold promise (Johnson e t a l . 1 9 7 8 ) , as d i f l u b e n z u r o n so a p p l i e d might w o r k . a s a c o n t a c t as w e l l as a stomach p o i s o n .  The compound has been  a p p l i e d a g a i n s t mosquitoes as a f l o w a b l e c o n c e n t r a t e and i n a g r a n u l a r form as w e l l as the w e t t a b l e powder ( M u l l a and Darwazeh  1976).  The t o x i c i t y o f d i f l u b e n z u r o n to n o n - t a r g e t organisms appears t o be r e l a t i v e l y low v i z . a c u t e o r a l ppm and  4,650  LC  5 Q  i(fish) =  LDgg ( r a t s ) =  135-195  to be n o n - t o x i c t o p r e d a t o r y m i t e s Kaldor  1980)  and n a t u r a l  4,640  ppm (Anonymous  mg/kg, 1974,  LC^Q  (birds) = I t appears  1977).  (Wearing and Thomas 1 9 7 8 ; Bower and  enemies o f the pear p s y l l a ( W e s t i g a r d  L i t t l e hazard i s p r e s e n t e d to A p i s m e l l i f e r a L.  (Anonymous  no a p p r e c i a b l e a c c u m u l a t i o n o f d i f l u b e n z u r o n i n b i o l o g i c a l ( M e t c a l f e t al_. 1 9 7 5 ) and MacGregor e_t al_. ( 1 9 7 9 )  1979).  1974).  There  food c h a i n s  observed no mutagenic  properties. In many w e l l - m a n a g e d , s a n i t a r y o r c h a r d s i n the Similkameen V a l l e y no  is  -8-  i n s e c t i c i d e s were r e q u i r e d throughout t h e growing season as l o n g as t h e c o d l i n g moth was c o n t r o l l e d by the s t e r i l i t y  program.  T h i s appears  highly  advantageous t o g r o w e r s , but a s e r i o u s d i f f i c u l t y may a r i s e as a r e s u l t . The r e d u c t i o n ' i n number o f i n s e e t i c i d a l sprays. i n - B r i t i s h Columbia o r c h a r d s because o f s t e r i l i t y - c o n t r o l  or integrated control  programs might  i n h i g h e r p o p u l a t i o n s o f a p p l e p e s t s not p r e v i o u s l y o f economic (Madsen and D a v i s 1971).  result  significance  T h i s was demonstrated by G l a s s and L i e n k  (1971)  who conducted a s t u d y on a New York a p p l e o r c h a r d , m a i n t a i n i n g i t f o r 10 y e a r s w i t h no i n s e e t i c i d a l ' s p r a y s .  W i t h i n two y e a r s , damage caused by the  c o d l i n g moth and a p p l e maggot reduced the economic v a l u e o f the crop to n i l . In a d d i t i o n to t h i s , p o p u l a t i o n s o f l e s s e r a p p l e p e s t s such as the plum c u r c u l i o , t h e l e s s e r appleworm and s e v e r a l s p e c i e s o f l e a f r o l l e r s T o r t r i c i d a e ) ' began t o  (Lepidoptera:  rise.  In the i n t e r i o r o f B r i t i s h Columbia t h e r e appear to be f o u r major s p e c i e s o f l e a f r o l l e r s which a t t a c k a p p l e . l e a f r o l l e r Archips argyrospilus  These i n c l u d e the  fruit-tree  ( W a l k e r ) , the European l e a f r o l l e r  rosanus ( L . ) , the t h r e e - l i n e d l e a f r o l l e r Pandemis l i m i t a t a obiiquebanded l e a f r o l l e r C h o r i s t o n e u r a rosaceana ( H a r r i s ) .  Archips  (Rob.) and the These f o u r  s p e c i e s a r e p e s t s on p e a r s , c h e r r i e s , a p r i c o t s and peaches as w e l l ejt al_. 1977; Madsen and Madsen 1980).  (Madsen  Other a p p l e - f e e d i n g l e a f r o l l e r s  are  p r e s e n t i n B r i t i s h Columbia but not as s i g n i f i c a n t pests (Doganlar and B e i r n e 1978, 1979).  A d u l t l e a f r o l l e r s a r e s m a l l , b e l l - s h a p e d moths, the  l a r v a e o f w h i c h f e e d on and r o l l  up l e a v e s o f v a r i o u s p l a n t s .  symptom p r o v i d e s p r o t e c t i o n a g a i n s t n a t u r a l c o n d i t i o n s and i n s e c t i c i d e s  This  foliar  enemies, adverse e n v i r o n m e n t a l  d u r i n g the l a r v a l and pupal s t a g e s .  can be e x t r e m e l y s e v e r e , but i n commercial a p p l e o r c h a r d s f r u i t  Defoliation injury  is  -9-  much more i m p o r t a n t than f o l i a r damage. developing f r u i t l e t s ,  Larvae feed i n buds and on young  c a u s i n g f l o w e r a b o r t i o n and damage to the young  The l a t t e r causes l a r g e u n s i g h t l y s c a r s t o develop as the f r u i t which r e s u l t s  fruit.  matures,  i n downgrading (Chapman 1973).  A- a r g y r o s p i l u s and A. rosanus a r e the most i m p o r t a n t p e s t s , p a r t i c u l a r l y the f o r m e r . their potential  While u s u a l l y o n l y marginal  leafroller problems,  s i g n i f i c a n c e i s c o n s i d e r a b l e , p a r t i c u l a r l y in orchards  where i n s e c t i c i d e use has been reduced by i n t e g r a t e d p e s t management o r sterility-control  f o r the c o d l i n g moth (Madsen 1970; Madsen and Davis 1971).  Both s p e c i e s o v e r w i n t e r i n egg masses and undergo a s i n g l e g e n e r a t i o n per y e a r (Madsen 1970). pink o r p e t a l - f a l l  A. a r g y r o s p i l u s and A. rosanus can be c o n t r o l l e d by a p p l i c a t i o n s of azinphos-methyl, d i a z i n o n or  (Madsen e t al_. 1977)..  P r e s e n t l y , growers a r e  trichlorfon  a d v i s e d to spray . a z i n p h o s -  methyl or d i a z i n o n a t p e t a l - f a l l , to reduce bee p o i s o n i n g (Anonymous 1980). P_. 1 i m i t a t a and C. rosaceana were o r i g i n a l l y c o n s i d e r e d to be u n i v o l t i n e i n the Okanagan' V a l l e y (Venables 1924; Mayer and B e i r n e 1974a,b) though P. 1 i m i t a t a was r e c o g n i z e d as b i v o l t i n e  i n Washington s t a t e  (Mayer  and B e i r n e 1974b) and C_. rosaceana the same i n New York s t a t e (Chapman e t al.  1968).  A t t h e p r e s e n t t i m e , both s p e c i e s appear t o be b i v o l t i n e  the Okanagan and Similkameen V a l l e y s , o v e r w i n t e r i n g as f i r s t - i n s t a r and a r e i n c r e a s i n g i n numbers and economic s i g n i f i c a n c e  in larvae,  ( H . F . Madsen, p e r -  sonal communication; R.D. M c M u l l e n , p e r s o n a l communication; Madsen and Madsen 1980).. During the 1979 growing s e a s o n , which was p a r t i c u l a r l y hot and d r y , p o p u l a t i o n s o f P.' 1 i m i t a t a and C_. rosaceana r o s e t o s e r i o u s l e v e l s and a number o f growers were f o r c e d to spray a g a i n s t the second g e n e r a t i o n .  -10-  A z i n p h o s - m e t h y l was g e n e r a l l y s e l e c t e d .  The problem was most s e r i o u s on  c h e r r i e s because the second brood emerges j u s t p r i o r to normal c h e r r y ting dates.  harves-  Sprays a r e not recommended a t t h i s t i m e , but the presence o f  l a r v a e and l a r v a l  s i l k on the f r u i t n e c e s s i t a t e s f u m i g a t i o n and/or  c l e a n i n g , a t c o n s i d e r a b l e expense.  careful  I f the problem p e r s i s t s , a chemical  o f low t o x i c i t y to h a r v e s t e r s which c o n t r o l l e d T e a f r o l l e r s e a r l y i n l i f e c y c l e s would be a boon t o c h e r r y growers ( R . D . M c M u l l e n ,  spray  their  personal  communication). I t can be seen t h a t l e a f r o l l e r s a r e s i g n i f i c a n t p e s t s i n the Okanagan and Similkameen V a l l e y s and t h a t the p o t e n t i a l o f d i f l u b e n z u r o n as a c o n t r o l agent i s worthy o f i n v e s t i g a t i o n .  F i e l d t e s t s have suggested t h a t  leaf-  r o l l e r s a r e not h i g h l y s e n s i t i v e to d i f l u b e n z u r o n (Wearing and Thomas 1978), but t h e s e were f i e l d  i n v e s t i g a t i o n s which d i d not e v a l u a t e a l l  to f i n d p o s s i b l e a r e a s o f s u s c e p t i b i l i t y .  life  stages  S e n s i t i v e l i f e stages s h o u l d be  c o n s i d e r e d when c h o o s i n g the t i m e , f o r m u l a t i o n and method o f a p p l i c a t i o n sprays.  A. a r g y r o s p i l u s and A. rosanus a r e d i f f i c u l t  i n s e c t s t o study  the l a b o r a t o r y because o f a tendency t o diapause every a l t e r n a t e  of  in  generation.  T h e r e f o r e , C_. rosaceana was s e l e c t e d as a r e p r e s e n t a t i v e t e s t s p e c i e s  of  Tortricidae. £• rosaceana i s a polyphagous i n s e c t which feeds on a l o n g l i s t u n r e l a t e d p l a n t s , but p r e f e r s woody Rosaceae. distributed  i n temperate North A m e r i c a .  of  I t i s n a t i v e t o and w i d e l y  C_. rosaceana appears to be u n i -  v o l t i n e i n c o l d e r Canadian a r e a s , b i v o l t i n e i n B r i t i s h Columbia and n o r t h e r n s t a t e s and t r i v o l t i n e i n Tennessee (Chapman e t al_. 1 9 6 8 ) . l a r g e s t and most fecund t o r t r i c i d s  I t i s one o f t h e  i n North America (Chapman and L i e n k 1 9 7 1 ) .  T h i s s p e c i e s was r e c o r d e d by Venables (1924) as the s i n g l e most  important  -11-  leafroller  i n t h e Okanagan V a l l e y p r i o r to 1918.  over t h e n e x t 60 y e a r s u n t i l difficulties 1973), i t  Its s i g n i f i c a n c e  t h e p r e s e n t r i s e i n numbers.  Considering  encountered i n the c l a s s i f i c a t i o n o f the T o r t r i c i d a e  i s possible that several  declined the  (Chapman  s p e c i e s have been i n v o l v e d w i t h i n the  same n o m e n c l a t u r e . R e i s s i g (1978) o b t a i n e d good c o n t r o l o f the obiiquebanded  leafroller  i n New York a p p l e o r c h a r d s w i t h methyl p a r a t h i o n and methomyl a p p l i e d a t petal-fall  and w i t h a z i n p h o s - m e t h y l , phosmet and methyl p a r a t h i o n  i n J u l y a g a i n s t the second g e n e r a t i o n . a d v i s e d to c o n t r o l  the f i r s t  applied  In B r i t i s h C o l u m b i a , growers are  (overwintering)  g e n e r a t i o n w i t h the same  measures used a g a i n s t the s i n g l e g e n e r a t i o n l e a f r o l l e r s , and t h e second (summer) g e n e r a t i o n w i t h d i a z i n o n (Anonymous 1980).  Orientation  disruption  o f a d u l t males u s i n g the s y n t h e t i c sex pheromone was i n s u f f i c i e n t t o p r o v i d e satisfactory control been improved ( H i l l  ( R e i s s i g e t aj_. 1978) but the s y n t h e t i c pheromones have and R o e l o f s 1979) and have e x c e l l e n t p o t e n t i a l  monitoring of population l e v e l s of t h i s  i n the  pest.  The i n t e n t o f t h i s study was t o e v a l u a t e the e f f e c t i v e n e s s o f d i f l u benzuron as a c o n t r o l agent f o r the c o d l i n g moth.  A second o b j e c t i v e was  to determine the s e n s i t i v i t y o f C_. rosaceana to the growth r e g u l a t o r . both c a s e s , l a b o r a t o r y s t u d i e s on the s e n s i t i v i t y o f d i f f e r e n t l i f e preceded f i e l d t r i a l s  i n Okanagan o r c h a r d s through a t y p i c a l  In  stages  growing s e a s o n .  MATERIALS AND METHODS A. R e a r i n g Techniques C o d l i n g moth eggs and a d u l t s were s u p p l i e d by r e s e a r c h e r s a t the A g r i c u l t u r e Canada Research S t a t i o n , Summerland, B.C. moths were r e q u i r e d f o r c e r t a i n t e s t s so s e v e r a l immature a p p l e s o f mixed c u l t i v a r s . 60% R.H. and 16-hour p h o t o p e r i o d .  Virgin adult  broods were r e a r e d  codling in  The c u l t u r e s were m a i n t a i n e d a t 25°C, A f t e r f e e d i n g i n the a p p l e s , mature  l a r v a e c r a w l e d i n t o and pupated w i t h i n c o r r u g a t e d wooden s t r i p s where they were c o l l e c t e d and s e x e d .  (Plate  I)  The sexed pupae were p l a c e d i n s e p a -  r a t e cages t o complete development to the a d u l t s t a g e .  Eggs were c o l l e c t e d  from a d u l t moths u s i n g a r o t a t i n g o v i p o s i t i o n cage l i n e d w i t h wax paper (Plate II)  ( a f t e r Proverbs and Logan 1970).  Egg masses o f the obiiquebanded l e a f r o l l e r were c o l l e c t e d from Okanagan o r c h a r d s by D r . H.F. Madsen.  A l a b o r a t o r y c o l o n y was e s t a b l i s h e d by  initi-  a l l y r e a r i n g the l a r v a e on an a g a r - b a s e d s y n t h e t i c d i e t composed o f the following constituents  ( m o d i f i e d a f t e r Redfern 1964; B r i n t o n e t aj_. 1969;  Bucher and Bracken 1976; M.D. P r o v e r b s , personal Vitamins: Agar c o n s t i t u e n t s :  communication):  Ascorbic acid Vanderzant v i t a m i n m i x t u r e Agar Casein Cysteine Wheat germ Wesson s a l t s W Choline chloride Methyl c e l l u l o s e Sucrose Soybean o i l 2 M K0H *Mold i n h i b i t o r D i s t i l l e d water  10.0 g 2.5 g 25.0 35.0 2.0 28.0 10.0 1.0 20.0 30.0 3.0 30.0 20.0 550.0  g g g g g g g g ml ml ml ml  -13-  *Mold i n h i b i t o r :  Sorbic acid Methyl parahydroxybenzoate Ethyl alcohol  20.0 g 15.0 g 170.0 ml  A f t e r m i x i n g the agar c o n s t i t u e n t s , the medium was heated to 90°C f o r 5 min and then c o o l e d t o 70°C, when t h e v i t a m i n m i x t u r e and a s c o r b i c a c i d were added.  F i v e t o 10 ml o f medium were p l a c e d i n j e l l y cups (Redfern 1964) o r  w h i t e p l a s t i c cream c o n t a i n e r s (Bucher and Bracken 1 9 7 6 ) .  The c o n t a i n e r s  were covered w i t h paper t o w e l l i n g t o reduce f u n g a l spore c o n t a m i n a t i o n and a l l o w e d t o s e t f o r 12-16 h t o a l l o w e x c e s s water t o e v a p o r a t e . t a i n e r s were then capped and r e f r i g e r a t e d a t - 4 ° C u n t i l sate f o r high m o r t a l i t y ,  The conr  required.  To compen-  two t o f o u r newly-emerged l a r v a e were p l a c e d i n  each c o n t a i n e r and r e a r e d a t c o n d i t i o n s d e s c r i b e d p r e v i o u s l y . method gave a poor y i e l d o f i n s e c t s as o n l y 10-15% o f the l a r v a e s u r v i v e d t o the a d u l t s t a g e .  This  rearing  first-instar  In a d d i t i o n t o t h i s , l a r v a l  development  l a s t e d f o u r t o s i x weeks w h i c h " n e c e s s i t a t e d t r a n s f e r r i n g the l a r v a e onto f r e s h medium two or t h r e e t i m e s .  C o n s e q u e n t l y , a f t e r t h r e e g e n e r a t i o n s on  the s y n t h e t i c d i e t , the l e a f r o l l e r s were r e a r e d e x c l u s i v e l y on broad beans ( V i c i a faba L.)  (Plate I I I ) .  The p l a n t s and a s s o c i a t e d l e a f r o l l e r s  were  r a i s e d i n a greenhouse, under a 16-hour p h o t o p e r i o d but w i t h v a r i a b l e temp e r a t u r e s (20-30°C) a n d r e l a t i v e  h u m i d i t y (70-95%).  Although  difficulties  encountered w i t h t h i s r e a r i n g method l i m i t e d the a v a i l a b i l i t y o f all  t e s t s were conducted on f o l i a r - f e d l e a f r o l l e r s .  Pupae were  insects, collected  from r o l l e d l e a v e s and p l a c e d i n wax paper cages to complete development t o the a d u l t stage ( P l a t e I V ) .  Egg masses were c o l l e c t e d from the i n n e r  f a c e s o f the c a g e s . B. T e s t Compounds Diflubenzuron ( D i m i l i n  25% W.P.; N - ( 4 - c h l o r o p h e n y l ) - N ' - ( 2 , 6 -  sur-  -14-  Plate I.  Plate  II.  Corrugated wooden s t r i p s used f o r c o l l e c t i o n o f c o d l i n g moth larvae.  R o t a t i n g o v i p o s i t i o n cage used f o r c o l l e c t i o n o f c o d l i n g moth eggs.  -15-  P l a t e IV.  Wax paper cage used f o r experiments on a d u l t moths and f o r c o l l e c t i o n of l e a f r o l l e r eggs.  -16-  difluorobenzoyl) Tween 20  u r e a ; Thompson-Hayward Chemical Company) and the  (polyoxyethylene  various concentrations million  surfactant  (20) s o r b i t a n monolaurate) were prepared a t  in d i s t i l l e d water.  The t e r m i n o l o g y " p a r t s per  (ppm)" i s used t o denote c o n c e n t r a t i o n . ,  These v a l u e s a r e  equivalent  t o micrograms o f compound per m i l l i l i t r e o f w a t e r . C. T e s t s on Eggs The c o n t a c t o v i c i d a l  a c t i v i t y o f d i f l u b e n z u r o n a n d / o r Tween 20 was  determined by a p p l y i n g the compound(s) t o c o d l i n g moth and l e a f r o l l e r u s u a l l y w i t h a f i n e " b r u s h . ' The e f f e c t s on egg hatch were r e c o r d e d u n t i l emergence w a s - c o m p l e t e . c a n n i b a l ism o f eggs.  eggs,  daily  Any hatched l a r v a e were-removed t o p r e v e n t  The s e n s i t i v i t y o f c o d l i n g moth eggs a t v a r y i n g  inter-  v a l s a f t e r o v i p o s i t i o n w a s t e s t e d by o b s e r v i n g the e f f e c t s o f - 1 0 ppm d i f l u benzuron on h a t c h a b i l i t y .  Paper t o w e l l i n g soaked i n 10 ppm d i f l u b e n z u r o n  was p l a c e d on c o d l i n g moth eggs f o r - d i f f e r e n t  lengthsof  time i n Order t o  determine the e f f e c t s o f d i f f e r e n t d r y i n g r a t e s on o v i c i d a l numbers o f r e p l i c a t e s  activity.  The  i n each s t u d y depended upon t h e a v a i l a b i l i t y o f eggs,  but each r e p l i c a t e u s u a l l y c o n s i s t e d o f 50 c o d l i n g moth eggs o r one  leaf-  r o l l e r egg-mass, which c o n t a i n e d c a . 50-600 eggs. The o v i c i d a l  a c t i v i t y o f d i f l u b e n z u r o n / T w e e n 20 a p p l i e d t o pear  and a p p l e s was a l s o a s s e s s e d .  The f o l i a g e ,  immature Golden D e l i c i o u s  foliage apples  and mature S p a r t a n a p p l e s were dipped i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s and a l l o w e d t o d r y .  Female c o d l i n g moths were a l l o w e d t o o v i p o s i t on the  t r e a t e d s u b s t r a t e s f o r f o u r to s i x days and the e f f e c t s on egg hatch r e corded a f t e r l a r v a l  e c l o s i o r i was c o m p l e t e .  o f d i f l u b e n z u r o n on f r u i t ,  To a s s e s s the r e s i d u a l  activity  immature Golden D e l i c i o u s a p p l e s were dipped i n a  s o l u t i o n . c o n t a i n i n g 100 p p m " d i f l u b e n z u r o n + 500 ppm Tween 20 and s t o r e d  in  cages f o r 10 days a t 25°C, .60% R.H. and 16-hour p h o t o p e r i o d .  A t t h e end o f  t h i s t i m e , female c o d l i n g moths were i n t r o d u c e d and a l l o w e d t o o v i p o s i t .  To  s i m u l a t e the e f f e c t s o f r a i n o r heavy dew on the e f f i c a c y o f d i f l u b e n z u r o n , pear f o l i a g e which had been d i p p e d i n 100 ppm d i f l u b e n z u r o n + 500 ppm Tween 20 was sprayed t o r u n - o f f w i t h d i s t i l l e d water on a d a i l y D  *  basis.  T e s t s on Larvae As young c o d l i n g moth l a r v a e have been observed t o f e e d on a p p l e  age p r i o r to e n t e r i n g t h e - f r u i t , t h e . t o x i c i t y o f d i f l u b e n z u r o n to i n s t a r l a r v a e was a s s e s s e d .  foli-  first-  Young a p p l e p l a n t s c a . 15 cm high were sprayed  to r u n - o f f w i t h d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s and a l l o w e d t o d r y .  First-  i n s t a r codl i n g m o t h . l a r v a e were p l a c e d i n c l i p - c a g e s which w e r e - a f f i x e d the f o l i a g e  (Plate V).  A f t e r the l a r v a e had f e d on the f o l i a g e f o r one o r  two days ( P l a t e V I ) , they were t r a n s f e r r e d onto a p p l e s .  The number o f  vae which s u c c e s s f u l l y e n t e r e d the apples and s u r v i v e d to the  F i r s t - i n s t a r l e a f r o l l e r l a r v a e were p l a c e d on t r e a t e d pear which had been dipped i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s . v a l was a s s e s s e d over two weeks.  Firsts  lar-  4th-instar  s t a g e was compared with- the s u c c e s s and development o f u n t r e a t e d  leafroller  to  larvae. foliage  Larval  survi-  2 n d - , 3 r d - , 4 t h - and 6 t h - i n s t a r  l a r v a e w e r e ' p l a c e d on t r e a t e d a p p l e f o l i a g e and t h e i r a b i l i t y  to  moult d e t e r m i n e d . E•  T e s t s on A d u l t s One- to t h r e e - d a y - o l d v i r g i n a d u l t c o d l i n g moths and l e a f r o l l e r s  dipped i n o r f e d s o l u t i o n s o f d i f l u b e n z u r o n and/or Tween 2 0 .  were  Four males  and females were p l a c e d i n wax paper cages and a l l o w e d t o d r i n k water o r t r e a t m e n t s o l u t i o n s from soaked c o t t o n w i c k s . and egg v i a b i 1 i t y were a s s e s s e d .  Adult longevity,  fecundity  P l a t e V.  Plate VI.  C l i p - c a g e used f o r t e s t s on f i r s t - i n s t a r c o d l i n g moth l a r v a e .  Apple f o l i a g e showing f e e d i n g i n j u r y by c o d l i n g moth instar larvae.  first-  -19-  F.  F i e l d -Tests  i)  C o d l i n g Moth All  f i e l d t e s t s were conducted a t t h e A g r i c u l t u r e Canada Research  S t a t i o n , Summerland.  The e f f e c t i v e n e s s o f s e v e r a l  diflubenzuron  formula-  t i o n s was • ' e v a l u a t e d a g a i n s t t h a t o f a z i n p h o s - m e t h y l , which was a p p l i e d a t recommended r a t e s .  The f i v e t r e a t m e n t s which were e v a l u a t e d i n c l u d e d 1)  unsprayed check, 2) 47 ppm d i f l u b e n z u r o n , 3) 47 ppm d i f l u b e n z u r o n + 500 ppm Tween 2 0 , 4) 187 ppm d i f l u b e n z u r o n and 5) 187 ppm a z i n p h o s - m e t h y l .  The  c o d l i n g moth t e s t s were-conducted i n an o r c h a r d o f mixed c u l t i v a r s and an o r c h a r d o f Golden D e l i c i o u s .  Both o r c h a r d s r e c e i v e d a s p r a y o f  on May 14, 1980 f o r c o n t r o l o f t h e San Jose s c a l e , perniciosus  (Comstock).  p l e t e block design  diazinon  Quadraspidaotus  In the Golden D e l i c i o u s o r c h a r d , a balanced incom-  ( H i c k s T973) was s e l e c t e d , • w i t h f i v e . b l o c k s , , f i v e  t r e a t m e n t s and f o u r " r e p l i c a t e s  per t r e a t m e n t ( F i g . l a ) .  s i s t e d o f t h r e e a d j a c e n t t r e e s o f the same e u l t i v a r , r e p l i c a t e s by a t l e a s t one " b u f f e r " t r e e .  Each r e p l i c a t e  separated  con-  from-other  In the m i x e d - c u l . t i v a r o r c h a r d ,  the check' t r e a t m e n t was o m i t t e d and i t s v a l u e i n f e r r e d by e x a m i n a t i o n -[oft b u f f e r t r e e s . • The r e m a i n i n g f o u r t r e a t m e n t s e a c h " c o n t a i n e d one r e p l i c a t e o f each e u l t i v a r  ( M c i n t o s h , ' S p a r t a n , D e l i c i o u s , Newtown and Rome), a r r a n g e d  i n a c o m p l e t e l y randomized d e s i g n ( F i g . l b ) .  Sprays were a p p l i e d a t * 21>  2 • kg/cm. u s i n g a c o n v e n t i o n a l  handgun s p r a y e r .  Spray dates were s e l e c t e d  to  c o i n c i d e w i t h t h e ' p e r i o d o f maximum f-1 i g h t a c t i v i t y which was determined w i t h the use o f Zoecon sex pheromone t r a p s (Myburgh e t ajk 1 9 7 4 ) . were a p p l i e d a g a i n s t t h e f i r s t petal-fall  Treatments  brood on May 28, 1980 (11 days a f t e r  peak  o f M c i n t o s h ) and a g a i n s t the second brood on August 7, 1980.  E f f e c t i v e n e s s o f the t r e a t m e n t s was a s s e s s e d on the b a s i s o f p e r c e n t  codling  -20-  Fig.  1.  Schematic r e p r e s e n t a t i o n . o f the c o d l i n g moth f i e l d - t e s t o r c h a r d , (0=buffer, • = c h e c k , »=47 ppm d i f l u b e n z u r o n , A=47 ppm d i f l u b e n zuron + 500 ppm Tween 2 0 , •=187 ppm d i f l u b e n z u r o n , H=187 ppm azinphos-methyl).  a)  Golden D e l i c i o u s  o  •  o  o  •  o  o  • • •  o  •  o  •  o  o  o  o  o  o  o  •  o  a  o  o •  o  •  o  •  o  B  o  •  o  •  o o  •  o o  IS  o  •  o  •  o  o  o  o  o  o  o o  •  o  •  o  o  •  •  o  •  o  •  o  o  o o  •  o  o  o  •  o  o  •  o o  •  o  •  o  a  B a  • •  b)  • • • o  o A  • A  o  o  o  o  BLOCK:  o  o  B B B  •  • • •  (J)  orchard.  o o  1  • • •  • •  o  •  o  • • •  o  •  o  o  o  o  •  o  B  •  o o  •  SI  •  o  •  o o  2  4  3  a 5  Mixed c u l t i v a r o r c h a r d .  o  O O  •  o  •  o  o  •  o  B B  o  •  o  B  O  A  o  o  o  o  O ' O  o  o  o o o o  o  •  o  • • •  o o o o  O  O  B B B  O  O O  o  • • •  o o o o  A  o  o  A  o  o  A  o  • • •  o  o  o  o  o o o o  o  o o o o  o  B B B  O  O 0 • • •  A A A  • • •  o  •  o  •  o  •  o  •  o  B B B  o  •  o  •  o  O  o  o  o  o  o o o o  o  o o o o  o  o o o o  o  o o  A A A  B B B  • • •  \  -21-  moth damaged f r u i t a t  harvest.  i i ) ' Mites Using methods d e s c r i b e d by Downing and A r r a n d ( 1 9 7 8 ) , phytophagous and predaceous mites were c o l l e c t e d from f o l i a g e throughout the summer i n both c o d l i n g moth t e s t o r c h a r d s .  O n ' t h e v a r i o u s - s a m p l i n g d a t e s , 50. l e a v e s were  c o l l e c t e d a t random f r o m f i v e t r e e s i n each t r e a t m e n t . brushed and the t o t a l  The l e a v e s were  numbers o f p r e d a t o r y and p e s t m i t e s e o u n t e d .  Within  each t r e a t m e n t , t h e same f i v e t r e e s were used throughout the summer f o r m i t e col l e c t i o n s . i i i ) • Leaf r o l l e r s  '  The t r e a t m e n t s t e s t e d " a g a i n s t l e a f r o l l e r s  .  i n c l u d e d 1) cheek, 2) 94 ppm  d i f l u b e n z u r o n , 3) 94 ppm d i f l u b e n z u r o n + 500 ppm Tween 2 0 , 4) 375 ppm d i f l u benzuron and 5) 375ppm a z i n p h o s - m e t h y l .  The'orchard selected f o r  this  s t u d y had r e c e i v e d few sprays i n r e c e n t y e a r s and c o n t a i n e d a m i x t u r e o f cultivars.  Because o f the wide host range o f l e a f r o l l e r s  (Madsen and Madsen  1 9 8 0 ) , d i f f e r i n g s u s c e p t i b i l i t y between c u l t i v a r s was not a n t i c i p a t e d and no attempt was made to i s o l a t e the c u l t i v a r s .  As i n the c o d l i n g moth t r i a l s ,  each r e p l i c a t e c o n s i s t e d o f t h r e e a d j a c e n t t r e e s , b u f f e r e d from o t h e r cates.  repli-  The f i v e t r e a t m e n t s were a r r a n g e d i n a randomized b l o c k d e s i g n w i t h  f i v e b l o c k s and f i v e r e p l i c a t e s per t r e a t m e n t .  (Fig. 2).  As 15-20  leafroller  l a r v a e were found per 100 l e a f c l u s t e r s , the i n f e s t a t i o n exceeded the economic t h r e s h o l d o f 10 l a r v a e / 1 0 0 c l u s t e r s spray was a p p l i e d on A p r i l  (Madsen e t al_. 1977).  A single  26, 1980 a t the p i n k bud stage o f M c i n t o s h .  t h i s t i m e , l e a f r o l l e r s were p r e d o m i n a t e l y a t the f i r s t - or 2 n d - i n s t a r E f f e c t i v e n e s s o f t r e a t m e n t s was a s s e s s e d on the b a s i s o f p e r c e n t damaged f r u i t a t  harvest.  At stage.  leafroller  ^22-  F i g . 2.  Schematic representation of the l e a f r o l l e r f i e l d - t e s t orchard. (-=dead or missing t r e e , 0=buffer t r e e , •=check, «=94 ppm diflubenzuron, *=94 ppm diflubenzuron + 500 ppm Tween 20, •=375 ppm diflubenzuron, B=375 ppm azinphos-methyl).  -  • •  \  •  \  •  •  \ •  0  \  m  a  A  o  a  \  \  \ t  0  A/  f  •  ;o  0  0  1  -  0  •  •  0  m «  0  . »  O  •  0  a 0  0  A O  0  A  • 0  0  * 4  0  •  0  •  0  •  0  Block 3 0 0 0  0  B O  0  B  0  f  f  l  O  . 0  0 «  •  •  0  A  0  Block 4 •  B  • 0  0 A  •  0  •  A  • 0  0  0  - - -  0 0  B  •  0  0  0  Block 2  0  A  •  •  •/ 0  Block 1 /  o  B  0  •  a  a o  •  0  f  •  - -  •  1 1  O  0  •  •  B  1 °  0  0  0  0  0 •>«•  -  •  •  -  0  •  •  -  A  0  1  /  0  -  j-  A  A  0  -  A  A  0 / 0 11  •  A  0  0  IB  \  \  0  ,'o  a  0  \  0  0  a  0  0  Block 5  0 0  0  -23-  G. S t a t i s t i c a l  Analysis  M u l t i p l e comparison procedures a r e used e x t e n s i v e l y and o f t e n rectly in s t a t i s t i c a l  a n a l y s i s of d a t a .  incor-  Such t e s t s s h o u l d o n l y be performed  when an experiment c o n t a i n s a l a r g e number o f q u a l i t a t i v e t r e a t m e n t s 1978).  Regression a n a l y s i s  statistical  s i g n i f i c a n c e of s e v e r a l  a regression is s i g n i f i c a n t , their effects, 1976).  all  determining  l e v e l s of a q u a n t i t a t i v e f a c t o r .  When  treatments are s i g n i f i c a n t l y d i f f e r e n t  i n c l u d i n g i n t e r m e d i a t e ones not used i n the experiment  Factorial  in  (Chew  a n a l y s i s o f v a r i a n c e t e s t s the e f f e c t s o f two o r more f a c -  tors simultaneously, other.  i s the a p p r o p r i a t e method f o r  (Little  i n c l u d i n g i n t e r a c t i o n e f f e c t s o f the f a c t o r s on each  When t r e a t m e n t s a r e q u a l i t a t i v e  i n n a t u r e , the a n a l y s i s o f  variance  w i t h p a r t i t i o n e d degrees o f freedom p r o v i d e s a powerful t e s t o f much g r e a t e r r e l i a b i l i t y than m u l t i p l e comparison procedures (Chew 1976, 1980). Experiments i n v o l v i n g q u a n t i t a t i v e t r e a t m e n t s were t e s t e d u s i n g r e g r e s s i o n a n a l y s i s and the method o f o r t h o g o n a l 1978).  polynomials  Q u a l i t a t i v e t r e a t m e n t s were a n a l y z e d u s i n g i n d i v i d u a l  .freedom.contrasts.and.factorial tested.simultaneously.(Li  1964).  The a n a l y s e s o f v a r i a n c e o f the above  i n d i c a t e d s i g n i f i c a n c e a t the 1% l e v e l .  graphical  p l o t s and maximum l i k e l i h o o d t e s t s  The term " h i g h l y  Probit analyses  signi-  involving  ( F i n n e y 1962) were used to  d e t e r m i n e LD^Q v a l u e s f o r c o d l i n g moth eggs, f i r s t - i n s t a r instar larvae.  degree o f  a n a l y s i s was conducted when two f a c t o r s were  t e s t s were conducted a t the 5% s i g n i f i c a n c e l e v e l . ficant"  ( L i t t l e and H i l l s  l a r v a e and 2nd-  To c o r r e c t f o r n a t u r a l m o r t a l i t y , A b b o t t ' s f o r m u l a was  a p p l i e d to the data p r i o r t o p r o b i t a n a l y s i s  (Abbott 1925).  The data o b -  t a i n e d were condensed and t a b u l a t e d i n the R e s u l t s s e c t i o n .  The raw d a t a  and s t a t i s t i c a l  a n a l y s e s were p l a c e d i n c o r r e s p o n d i n g t a b l e s i n the A p p e n d i x .  -24-  RESULTS I.  Laboratory  Studies  A.  C o d l i n g Moth  i)  Ovicide  Tests  Table I shows the c o n t a c t o v i c i d a l  a c t i v i t y of diflubenzuron  t o p i c a l l y w i t h a f i n e brush to eggs o f d i f f e r e n t ages.  When d i f l u b e n z u r o n  was a p p l i e d to 3 - d a y - o l d e g g s , egg hatch was o n l y s l i g h t l y ppm but was s e v e r e l y reduced a t 100 ppm. oviposition)  applied  i n h i b i t e d a t one  Younger eggs (0-60 hours  showed i n c r e a s e d s e n s i t i v i t y to d i f l u b e n z u r o n .  after  Although a r e -  d u c t i o n i n egg hatch was apparent a t 0.1 ppm, 100 ppm d i f l u b e n z u r o n o v e r 95% s u p p r e s s i o n o f egg h a t c h . (Abbott 1925) g r a p h i c a l  After correcting for natural  produced  mortality  p l o t s and maximum l i k e l i h o o d t e s t s were performed  to determine L D ^ v a l u e s f o r young and o l d eggs ( T a b l e I I ) .  The LD  values  o b t a i n e d suggested g r e a t e r s e n s i t i v i t y o f younger eggs to d i f l u b e n z u r o n . such i s the c a s e , the LD  v a l u e f o r c o d l i n g moth eggs would v a r y  a b l y depending on the t i m i n g o f  If  consider-  treatment.  As embryonic development i n the c o d l i n g moth c o l o n y was n o r m a l l y comp l e t e d w i t h i n seven d a y s , t h e s e n s i t i v i t y o f 0 - t o 1 3 2 - h - o l d eggs t o 10 ppm d i f l u b e n z u r o n was determined ( T a b l e I l i a ) .  T h i s c o n c e n t r a t i o n was chosen  because i t produced i n t e r m e d i a t e y e t s i g n i f i c a n t egg m o r t a l i t y .  Regression  a n a l y s i s showed a h i g h l y s i g n i f i c a n t e f f e c t o f egg age on s e n s i t i v i t y diflubenzuron.  Younger eggs were much more s e n s i t i v e , e s p e c i a l l y up t o  60 h p o s t o v i p o s i t i o n . reduced s t i l l  to  further,  When the time between o v i p o s i t i o n and t r e a t m e n t was i t was e v i d e n t t h a t eggs were more s e n s i t i v e  d i f l u b e n z u r o n when t r e a t e d i m m e d i a t e l y a f t e r o v i p o s i t i o n  to  ( T a b l e 11 l b ) .  -25-  Table I.  P e r c e n t hatch and c o n t r o l o f c o d l i n g moth eggs t r e a t e d w i t h varying concentrations of d i f l u b e n z u r o n .  Egg age (days a f t e r oviposition) 3-day-old  Diflubenzuron cone. (ppm)  topically  % hatch (mean i S . E . )  % control  0  78.5 ± 1.7  -  1.0  69.0 ± 2.6  13.2  10.0  47.5 ± 5.1  40.3  100.0  18.5 ± 1.5  76.7  2  (Linear regression highly s i g n i f i c a n t , quadratic regression s i g n i f i c a n t ) 0  68.7 ± 2.5  -  0.01  70.4 - 2.4  0  0.1  50.0 ± 2.5  28.1  1.0  39.3 ± 2.0  43.5  10.0  , 8 . 9 ± 2.1  87.2 •  2i-day-old  100.0 (Linear regression highly c o r r e c t e d a f t e r Abbott 50 e g g s / r e p l i c a t e ,  4.9 t significant)  (1925).  four  replicates/treatment.  50 e g g s / r e p l i c a t e , n i n e  replicates/treatment.  1.1  92.9  -26-  Table I I .  Maximum l i k e l i h o o d t e s t s on the t o x i c e f f e c t s o f to eggs o f the c o d l i n g moth.  Parameter No. e g g s / t r e a t m e n t  diflubenzuron  3 - d a y - o l d eggs  2^-day-old eggs  200  450  Check m o r t a l i t y ± S . E .  21.5 ± 1 . 7  31.3*2.5  Probit l i n e slope ± S.E.  0.95 ± 0.11  0.82 ± 0.05  17.2  1.1  12.1 - 24.6  0.4 - 3.2  LD  5 Q  (ppm)  95% c o n f i d e n c e l i m i t s Heterogeneity f a c t o r  (X ) 2  0.32  16.1  -27-  Table I I I .  P e r c e n t hatch o f c o d l i n g moth eggs t r e a t e d t o p i c a l l y w i t h 10 ppm d i f l u b e n z u r o n a t v a r y i n g times from o v i p o s i t i o n .  Egg age (hours a f t e r oviposition) a)  1  % hatch (mean 15.6  0 - 36  2  i 2.9  36 - 60  18.0 - 2.3  60 - 84  41.3 ± 3.7  84 - 108  39.8  t 3.5  108 - 132  61.1  - 4.9  (Linear-regression highly b)  S.E.)  significant) 5.0 - 2.9  0 - 12 12 - 24  40.0  24 - 36  32.5 - 9.5  36 - 48  60.0 (Linear regression  significant)  50 e g g s / r e p l i c a t e , n i n e  replicates/treatment.  10 e g g s / r e p l i c a t e , f o u r  replicates/treatment.  i 4.1  t 9.1  -28-  In the p r e v i o u s t e s t s , d i f l u b e n z u r o n s o l u t i o n s remained i n c o n t a c t w i t h the eggs f o r upwards o f f o u r hours b e f o r e d r y i n g .  When d r y i n g was  a c c e l e r a t e d with a fan in p r e l i m i n a r y t e s t s , h a t c h a b i l i t y i n c r e a s e d . t h i s r e a s o n , t e s t s were performed t o determine the i n f l u e n c e o f r a t e s on the o v i c i d a l  a c t i v i t y of diflubenzuron.  drying  As i n d i c a t e d i n T a b l e  IVa d i f l u b e n z u r o n was more e f f e c t i v e as an o v i c i d e when kept i n on t h e egg s u r f a c e s f o r l o n g e r p e r i o d s o f t i m e .  For  solution  In t h i s t e s t , the time i n  s o l u t i o n was m a n i p u l a t e d by b l o t t i n g the eggs d r y w i t h paper t o w e l l i n g . However, w i t h t h i s method i t was f e l t t h a t  l e s s : diflubenzuron  would be l e f t on the eggs than i f they d r i e d n o r m a l l y . t e s t was r e p e a t e d , e x c e p t i n t h i s i n s t a n c e  residue  Consequently,  the  time i n s o l u t i o n was e n f o r c e d  by p l a c i n g d i f 1 u b e n z u r o n - s o a k e d paper t o w e l l i n g i n c o n t a c t w i t h t h e eggs f o r v a r y i n g i n t e r v a l s and then a l l o w i n g t h e eggs to d r y . ( T a b l e I V b ) . egg hatch was s i g n i f i c a n t l y reduced as time i n s o l u t i o n  Again,  increased.  The s u r f a c t a n t Tween 20 was used i n l a b o r a t o r y s t u d i e s t o reduce the s u r f a c e t e n s i o n o f t e s t s o l u t i o n s and improve d i s t r i b u t i o n and r e t e n t i o n on various surfaces.  The presence o f Tween 20 a p p a r e n t l y enhanced the o v i c i d a l  a c t i o n o f d i f l u b e n z u r o n s o l u t i o n s a p p l i e d d i r e c t l y to 3 - d a y - o l d and 1 t o 2 d a y - o l d eggs ( T a b l e V ) . Mature a p p l e s dipped i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s showed s i g n i ficant ovicidal  activity,  but not e f f e c t i v e c o n t r o l  (Table V i a ) .  These  a p p l e s were e x t r e m e l y waxy, however, and s o l u t i o n s tended t o bead up and drop o f f even when Tween 20 was added.  When immature a p p l e s were u s e d ,  which were much l e s s waxy, the r e s i d u a l o v i c i d a l was e x c e l l e n t , residual  action of  diflubenzuron  p a r t i c u l a r l y when Tween 20 was i n c l u d e d ( T a b l e V I b ) .  a c t i v i t y was unreduced 10 days a f t e r a p p l i c a t i o n .  Residual  This control  -29-  T a b l e IV.  P e r c e n t hatch o f c o d l i n g moth eggs p l a c e d i n c o n t a c t w i t h 10 ppm d i f l u b e n z u r o n s o l u t i o n s f o r varying periods of time.  Time i n solution (hours) a)  1  % hatch (mean -  S.E.)  0.25  77.5 i  1.3  0.5  69.5 -  1.3  1.0  71.5 t  1.5  2.0  61.5  4.0  40.5 - 3.1  t 1.3  (Log 1 i n e a r r e g r e s s i o n h i g h l y s i g n i f i c a n t , l o g q u a d r a t i c and l o g c u b i c r e g r e s s i o n s s i g n i f i c a n t ) b)  2  0.25  76.0 ± 3.6  0.5  64.2 - 2.8  1.0  60.2 ± 2.6  2.0  58.4  4.0  36.7 - 7.6  t 1.8  (Log l i n e a r r e g r e s s i o n h i g h l y s i g n i f i c a n t , l o g q u a d r a t i c and l o g c u b i c r e g r e s s i o n s s i g n i f i c a n t ) 50 e g g s / r e p l i c a t e , f o u r r e p l i c a t e s / t r e a t m e n t ,  eggs b l o t t e d d r y .  50 e g g s / r e p l i c a t e , n i n e r e p l i c a t e s / t r e a t m e n t , c o n t a c t w i t h soaked paper t o w e l l i n g .  eggs d r i e d n o r m a l l y  after  T a b l e V.  Treatment  a)  1  Percent hatch o f c o d l i n g moth eggs t r e a t e d w i t h d i f l u b e n z u r o n i n the presence (+) and absence (-) of Tween 20.  500 ppm Tween 20  1 ppm diflubenzuron  B  +  -  E  -  -  • F..'  +  Contrasts :  b)  2  10 ppm diflubenzuron  solutions  % ha£ch (mean - S . E . ) 82.0  +  1.8  +  40.5  +  3.0  +  17.0  +  2.1  +  1.6  (B+E) versus (F) - h i g h l y s i g n i f i c a n t , (B) versus (E) - h i g h l y s i g n i f i c a n t .  A  -  -  76.0  B  +  -  76.4  C  -  +  68.9  D  +  +  63.3  E  -  -  +  40.0  F  +  -  +  32.7  F a c t o r i a l ANOVA:  effects effects  50 e g g s / r e p l i c a t e , f o u r r e p l i c a t e s / t r e a t m e n t ,  + + + + +  1.8 2.5 2.5 3.0 2.9  of diflubenzuron - h i g h l y s i g n i f i c a n t . o f Tween 20 - s i g n i f i c a n t .  eggs 3 days o l d a t time of  treatment.  50 e g g s / r e p l i c a t e , nine r e p l i c a t e s / t r e a t m e n t , eggs 1-2 days o l d a t time o f  treatment.  Table V I .  H a t c h a b i 1 i t y o f c o d l i n g moth eggs o v i p o s i t e d on a p p l e s which had been dipped i n diflubenzuron/Tween 20 s o l u t i o n s .  Treatment a)  500 ppm Tween 20.  100 ppm diflubenzuron  A  % hatch (mean - S . E , 8 1 . 0 ± 1.7  B  +  75;3 - 2.1  C  +  73.0 -  Contrasts:  (A) v s . (B+C) - s i g n i f i c a n t . (B) v s . (C) - not ' s i g n i f i c a n t . 67.3 -  b)'  +  Contrasts: - -  ...  1.5'  1.8  +  40.2 - 2.1  +  21.2  t 1.5  19.5  t 1.2  (A) v s . (B+C+D) - h i g h l y s i g n i f i c a n t . (B) v s . (C+D) - h i g h l y s i g n i f i c a n t . ,(C) v s . (D) - not s i g n i f i c a n t .  1 s i x mature Spartan apples per t r e a t m e n t . •12 immature Golden D e l i c i o u s a p p l e s per t r e a t m e n t . t h e a p p l e s i n t h i s treatment were t r e a t e d 10 days e a r l i e r than the o t h e r s .  -32-  o f eggs was a l s o good on pear f o l i a g e  (Table V i l a )  but was even b e t t e r  the l e a v e s were l e f t on twigs p l a c e d i n water ( T a b l e V l l b ) , as t h e s e remained t u r g i d and h e a l t h y much l o n g e r .  The r e w e t t i n g t r e a t m e n t ,  s i m u l a t e d r a i n o r heavy dew, decreased the r e s i d u a l diflubenzuron 1 1  )  ( T a b l e V 1 1 b ) , but a p p a r e n t l y not  ovicidal  if  leaves  which  activity  of  significantly.  L a r v i c i d e Tests . First-instar  l a r v a e showed high m o r t a l i t y when f e d agar medium c o n -  taining diflubenzuron/(Table V i l l a ) . more s u s c e p t i b l e  ( T a b l e V11 l b ) .  S e c o n d - i n s t a r l a r v a e appeared even  M o r t a l i t y o c c u r r e d d u r i n g the m o u l t ;  f e c t e d l a r v a e were unable to shed t h e i r e x u v i a e o r formed t h i n  af-  cuticles  which r u p t u r e d e a s i l y .  P r o b i t a n a l y s i s was conducted to determine LD^Q  v a l u e s f o r both i n s t a r s  (Table I X ) .  s e n s i t i v e than the f i r s t - i n s t a r First-instar  The 2 n d - i n s t a r l a r v a e appeared more  larvae.  l a r v a e which had been f e d d i f l u b e n z u r o n - t r e a t e d  f o l i a g e f o r 1-2 days were p l a c e d on a p p l e s .  apple  The v a r i o u s t r e a t m e n t s had no  s i g n i f i c a n t e f f e c t on the a b i l i t y o f the l a r v a e to p e n e t r a t e the a p p l e s (Table Xa).  However, both d i f l u b e n z u r o n c o n c e n t r a t i o n and time o f  were found t o be s i g n i f i c a n t f a c t o r s (Table Xa).  i n the subsequent s u r v i v a l  of  feeding larvae  S u r v i v a l was a s s e s s e d o n l y t o the 4 t h - i n s t a r because o f  d e t e r i o r a t i o n of apples.  rapid  While most l a r v a e s u c c e s s f u l l y e n t e r e d a p p l e s ,  the m a j o r i t y o f those which f e d on 500 ppm d i f l u b e n z u r o n - t r e a t e d d i d not moult s u c c e s s f u l l y i n t o 2 n d - i n s t a r s . a p p l e s , l e a v i n g a small b l e m i s h ( " s t i n g " ) .  foliage  They d i e d c a . 8 mm i n s i d e  the  Newly emerged l a r v a e which were  p l a c e d on sprayed a p p l e s w i t h o u t p r i o r f e e d i n g on d i f l u b e n z u r o n - t r e a t e d f o l i a g e s u c c e s s f u l l y e n t e r e d and f e d on the f r u i t w i t h no apparent in mortality  (Table Xb).  increase  Table V I I .  a)  H a t c h a b i l i t y o f c o d l i n g moth eggs o v i p o s i t e d on pear f o l i a g e which had been dipped i n diflubenzuron/Tween 20 s o l u t i o n s .  Treatment  500 ppm Tween 20  100 ppm diflubenzuron  A  +  -  79.9 -  B  +•  +  36.1 - 3.3  Contrast: b)'  (A) v s . (B) - h i g h l y  C  94.3 - 2.2 +  48.1 - 9.5  +  +  26.7  +  +  40.1 ± 7.6  Contrasts:  2  3  s i x leaves/treatment,  n i n e leaves/treatment,  2.2  significant.  A B  1  % hatch (mean - S . E . )  (A) v s . (B+C+D) - h i g h l y s i g n i f i c a n t . (B) v s . (C+D) - not s i g n i f i c a n t . (C) v s . (D) - not s i g n i f i c a n t .  leaves not s u s t a i n e d by w a t e r . l e a v e s a t t a c h e d to t w i g s placed i n w a t e r .  t h e f o l i a g e i n t h i s treatment was rewetted d a i l y w i t h d i s t i l l e d  water.  -'4.5  Table V I I I .  M o r t a l i t y -Of a) f i r s t - and b) 2 n d - i n s t a r c o d l i n g moth l a r v a e f e e d i n g on agar medium c o n t a i n i n g v a r i o u s c o n c e n trations of diflubenzuron. ;  Diflubenzuron cone. (ppm) a)  2  3  (mean ± S . E . )  0  15.0  +  5.0  io'  45.0  +  9.6  50  50.0  +  5.8  100  70.0  +  5.8  0  20.0  +  8.2  10  65.0  +  9.6  50  85.0  +  5.0  100  95.0  +  5.0  (Linear regression b)  % mortality  significant)  (Linear regression s i g n i f i c a n t , quadratic regression s i g n i f i c a n t ) ^modified from Anderson ( 1 9 7 8 ) . 2  f i v e f i r s t - i n s t a r l a r v a e per r e p l i c a t e ( j e l l y c u p ) , 4 r e p l i c a t e s / t r e a t m e n t , m o r t a l i t y a s s e s s e d a f t e r 14 d a y s . f i v e 2 n d - i n s t a r l a r v a e per r e p l i c a t e ( j e l l y c u p ) , 4 r e p l i c a t e s / t r e a t m e n t , m o r t a l i t y a s s e s s e d a f t e r 10 d a y s .  -35-  T a b l e IX.  Maximum l i k e l i h o o d t e s t s on the t o x i c e f f e c t s o f to l a r v a e o f the c o d l i n g moth.  Parameter  lst-instar  No. l a r v a e / t r e a t m e n t  larvae  diflubenzuron  2nd-instar  larvae  20  20  Check m o r t a l i t y - S . E .  15.0 ± 5.0  20.0 - 8.2  Probit l i n e slope ± S.E.  0.69 - 0.48  1.28 ± 0.53  LD  5 Q  (ppm)  48.2  95% c o n f i d e n c e l i m i t s Heterogeneity f a c t o r  13.6-171.0 C* ) 2  1.10  8.13 2.2-30.0 0.38  T a b l e X.  P e n e t r a t i o n o f apples and s u r v i v a l to 4 t h - i n s t a r by c o d l i n g moth f i r s t - i n s t a r l a r v a e , a f t e r f e e d i n g on a p p l e f o l i a g e sprayed w i t h d i f l ubenzuron/Tween 20.  Diflubenzuron cone. (ppm)  500 ppm Tween 20  days feeding  % entering apples (mean ± S . E . )  0  -  1 2  87.5 ± 4.8 80.0 ± 4.1  65.0 - 5.0 65.0 ± 6.5  0  +  1 2  85.0 t 2.9 87.5 ± 4.8  70.0 t 4.1 65.0 ± 2.9  10  +  1 2  85.0 t 2.9 . 82.5 ± 4.8  65.0 ± 5.0 60.0 ± 4.1  100  +  1 2  85.0 ± 5.0 70.0 ± 7.1  70.0 ± 4.1 52.5 - 2.5  500  +  1 2  85.0 ± 2.9 77.5 ± 4.8  27.5 ± 4.8 15.0 ± 6.5  F a c t o r i a l ANOVA (on % e n t e r i n g a p p l e s ) : (on s u r v i v a l b)  2  to 4 t h - i n s t a r ) :  between t r e a t m e n t s .  e f f e c t of diflubenzuron - highly s i g n i f i c a n t . e f f e c t o f time f e e d i n g - s i g n i f i c a n t .  0  +  0  75.0  60.0  500  +  0  80.0  60.0  ' t e n l a r v a e and a p p l e s / r e p l i c a t e , f o u r 2  no s i g n i f i c a n t d i f f e r e n c e s  % survival to 4 t h - i n s t a r (mean ± S . E . )  2 0 l a r v a e and a p p l e s / t r e a t m e n t . without f o l i a r feeding.  replicates/treatment.  Newly-emerged l a r v a e were p l a c e d on sprayed a p p l e s  -37-  1  i )  T o x i c and c h e m o s t e r i l a n t t e s t s on a d u i t s  1  The t o x i c and c h e m o s t e r i l a n t p r o p e r t i e s were i n i t i a l l y d i p p i n g the a d u l t s  in s o l u t i o n s c o n t a i n i n g varying concentrations of  benzuron (Table X I ) .  L o n g e v i t y was not s i g n i f i c a n t l y reduced by the  ments, a l t h o u g h males l i v e d l o n g e r than f e m a l e s . unaffected.  a s s e s s e d by diflutreat-  Egg v i a b i l i t y was a l s o  S i n c e the number o f eggs l a i d c o u l d be a f f e c t e d by l o n g e v i t y  o f f e m a l e s , f e c u n d i t y was a s s e s s e d and a n a l y z e d i n terms o f eggs/cage and eggs/female/day.  In both i n s t a n c e s , r e g r e s s i o n a n a l y s i s  r e d u c t i o n i n f e c u n d i t y due to d i f l u b e n z u r o n t r e a t m e n t s .  showed  significant  When these  tests  were r e p e a t e d w i t h Tween 2 0 / d i f l u b e n z u r o n m i x t u r e s , f e c u n d i t y was not r e duced ( T a b l e X I I ) . fecundity,  The t r e a t m e n t s had no e f f e c t on a d u l t l o n g e v i t y or  but egg hatch was s i g n i f i c a n t l y reduced a t 100 ppm d i f l u b e n z u r o n  as compared to 10 ppm.  A d u l t moths f e d d i f l u b e n z u r o n s o l u t i o n s  showed no  r e a c t i o n i n terms o f l o n g e v i t y o r f e c u n d i t y , but egg v i a b i l i t y was s i g n i ficantly  reduced ( T a b l e X I I I ) .  B.  Obiiquebanded  i)  Ovicide Tests  Leafrol1er  The egg masses o f the o b i i q u e b a n d e d l e a f r o l l e r poor hatch even i n the checks ( T a b l e X I V ) . and i t  showed a c o n s i s t e n t l y  Mold was one reason f o r  i s p o s s i b l e t h a t poor f e r t i l i z a t i o n was a l s o r e s p o n s i b l e .  this,  This  c o m p l i c a t e d i n v e s t i g a t i o n s on the s u s c e p t i b i l i t y o f egg masses to d i f l u b e n zuron.  I t was a p p a r e n t , however, t h a t s u s c e p t i b i l i t y to d i f l u b e n z u r o n was  low s i n c e even 1,000 ppm d i f l u b e n z u r o n had no d e t e c t a b l e i n h i b i t o r y on egg h a t c h .  R e g r e s s i o n a n a l y s i s showed no e f f e c t ,  which showed no hatch were e x c l u d e d from a n a l y s i s .  influence  even when egg masses D e s p i t e the presence of  a conspicuous d e p o s i t o f d i f l u b e n z u r o n on the egg masses, the l a r v a e which  Table X I .  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t c o d l i n g moths dipped i n diflubenzuron solutions.  Diflubenzuron cone. (ppm)  Male longevity (days)  Female longevity (days)  (eggs/cage)  Ceggs/J/day)  0  15.2 ± 1 . 3  9.7 ± 1.6  521.3 ± 1 1 7 . 7  14.6 ± 1 . 9  65.9 ± 8 . 8  10  8.9 ± 1.5  9.1 - 1 . 9  248.3 ± 56.3  7.0 ± 0 . 8  69.4 ± 9 . 1  100  9.2 ± 1 . 3  6.8 ± 1 . 5  194.7 ± 13.9  7.4 ± 1 . 0  65.9 ± 9 . 1  1  Fecundity*  Fecundity*  * L i near r e g r e s s i on s s i gni f i cant f o u r males and f e m a l e s / c a g e , t h r e e  cages/treatment.  % hatch  Table X I I .  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y of a d u l t c o d l i n g moths dipped i n diflubenzuron/Tween 20 s o l u t i o n s .  Diflubenzuron cone. (PPm)  500 ppm Tween 20  1  Male longevity  Female longevity  (days)  (days)  Fecundity (eggs/cage)  (A)  0  12.0 + 1.1  9.3 ± 1 .3  550.1  (B)  10  8.6 + 1.2  9.2 i 1 .0  (C)  100  1.5  9.8 ± 0 .8  (D)  100  7.8 + 1.0  10.1 i 1 .1  +  9.3 +  *Contrasts: f o u r males and f e m a l e s / c a g e , t h r e e  t 1.8  Fecundity (eggs/g/day) 14.6 ± 4 . 2  75.6  418.7 ± 4 7 . 9  11.3 ± 2.9  84.6  546.3 ± 53.2  13.8 - 3.5  75.3  374.3 ± 156.2  11.2  t 4.7  64.4  % hatch of (B) s i g n i f i c a n t l y g r e a t e r than (C+D).  cages/treatment.  % hatch*  +  +  +  +  6.5 2.7 2.5 6.0  Table X I I I .  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t c o d l i n g moths f e d diflubenzuron solutions.  Diflubenzuron cone, (.ppm)  Male longevity (days)  Female longevity (days)  (eggs/cage)  (eggs/g/day)  0  12.0 ± 0.9  10.1 ± 0.7  514.7 ± 35.5  13.2 ± 2.0  81.4 ± 3.0  10  11.8 ± 0 . 8  11.5 - 1.0  563.3 ± 129.1  12.1 ± 2 . 3  79.9 ± 3.1  100  11.6 ± 0 . 9  11.7 ± 1.0  603.0 ± 27.6  12.9 ± 0.6  61.6 ± 7 . 3  1  '  :  f o u r males and f e m a l e s / c a g e , t h r e e  Fecundity  * Linear regression cages/treatment.  Fecundity  significant  % hatch*  -41-  T a b l e XIV.  P e r c e n t hatch o f l e a f r o l l e r egg masses t o p i c a l l y t r e a t e d w i t h varying concentrations of d i f l u b e n z u r o n .  Diflubenzuron cone. (ppm)  % hatch  (mean - S . E . )  0  22.9 ± 14.0  57.2  +  1.0  • 1,  37.9 - 17.5  63.1  +  15.0  10  33.9 - 16.5  42.4  +  18.3  100  62.1 ± 15.8  77.6  1,000  36.0 - 15.2  60.0  regressions all  1  % hatch^ (mean ± S . E . )  +  +  3.8 7.1  not s i g n i f i c a n t  egg masses i n c l u d e d i n the c a l c u l a t i o n s ; f i v e  masses/treatment.  those egg masses which showed no hatch were excluded from the calculations.  -42-  emerged were a p p a r e n t l y h e a l t h y and e x h i b i t e d no subsequent i l l ii)  Larvicide  effects.  Tests  When f i r s t - i n s t a r  leafroller  l a r v a e were r e a r e d on pear f o l i a g e  that  had been dipped i n v a r y i n g c o n c e n t r a t i o n s o f d i f l u b e n z u r o n and/or Tween 2 0 , larvicidal  e f f e c t s were apparent ( T a b l e X V ) .  i n the check t r e a t m e n t , f a c t o r i a l  a n a l y s i s o f v a r i a n c e found  i n t e r a c t i o n o f d i f l u b e n z u r o n w i t h Tween 20. a l o n e were not s i g n i f i c a n t ,  D e s p i t e high l a r v a l  The e f f e c t s of  significant diflubenzuron  i n d i c a t i n g t h e importance o f the s u r f a c t a n t  enhancing the a c t i o n o f d i f l u b e n z u r o n .  The m o u l t - i n h i b i t i n g p r o p e r t i e s  d i f l u b e n z u r o n / T w e e n 20 were assessed on the l a r v a l  instars  s u s c e p t i b i l i t y was a p p a r e n t .  Differences  in of  (Table XVI).  though the l i m i t e d a v a i l a b i l i t y o f i n s e c t s p r e c l u d e d s t a t i s t i c a l larval  mortality  Al-  analysis,  in sensitivity  between i n -  5  s t a r s c o u l d not be d e t e r m i n e d . iii)  T o x i c and c h e m o s t e r i l a n t t e s t s on a d u l t s A d u l t l e a f r o l l e r moths dipped i n 100 ppm d i f l u b e n z u r o n showed a s i g n i -  f i c a n t r e d u c t i o n i n l o n g e v i t y when compared to those dipped i n 0 ppm d i f l u benzuron ( T a b l e X V I I ) .  No r e d u c t i o n was observed i n f e c u n d i t y o r egg  viabi1ity. II. A.  F i e l d Tests Leafrol1er  test  The seasonal abundance o f f o u r l e a f r o l l e r i s shown i n F i g . 3.  s p e c i e s i n the t e s t  orchard  The t w o - g e n e r a t i o n l e a f r o l l e r s , p a r t i c u l a r l y C_.  r o s a c e a n a , were the most abundant t o r t r i c i d s , as i n d i c a t e d by the c a t c h e s o f sex pheromone t r a p s .  T r a p p i n g was c u r t a i l e d i n m i d - A u g u s t ,  the c a p t u r e d moths were l i k e l y f i r s t - g e n e r a t i o n a d u l t s Madsen 1980).  consequently  (see Madsen and  T a b l e X V I I I shows the p e r c e n t o f h a r v e s t e d a p p l e s  exhibiting  -43-  T a b l e XV.  P e r c e n t s u r v i v a l o f f i r s t - i n s t a r l e a f r o l l e r l a r v a e f e e d i n g on pear f o l i a g e which had been dipped i n d i f l u b e n z u r o n / T w e e n 20 solutions.  Diflubenzuron cone. (ppm)  500 ppm Tween 20  0  -  44.5 ± 1 1.1  0  +  88.9 ± 11.1  10  -  66.7 ± i 1.1  10  +  55.5 ± " 1.1  100  -  55.5 ± " 1.1  100  +  11.1 ± " 1.1  Factorial  ANOVA:  % survival  1  (mean ± S . E . )  d i f l u b e n z u r o n X Tween 20 i n t e r a c t i o n -  three l a r v a e / l e a f , three leaves/treatment, days o f f e e d i n g .  survival  significant.  a s s e s s e d a f t e r 14  Table XVI.  M o u l t i n g success o f f i v e l e a f r o l l e r l a r v a l i n s t a r s p l a c e d on appl f o l i a g e sprayed w i t h diflubenzuron/Tween 20 s o l u t i o n s . *  difleTuron  one l a r v a / p l a n t ,  T ^ 2 0  five  l  s  t  i  n  s  t  -  ^  instar  3rd i n s t a r  4th i n s t a r  6th  instar  plants/treatment.  i -Pi I  T a b l e "XVII.  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t l e a f r o l l e r moths dipped d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s .  100 ppm diflubenzuron  +  500 ppm 1  Tween 20  Male longevity* (days)  Female longevity* (days)  +  5.8 t 0.6  +  4 . 3 ± 0.6 * Contrast:  f o u r males and f e m a l e s / c a g e , f o u r  Fecundity  Fecundity  in  % hatch  (eggs/cage)  (eggs/^/day)  6.2 t 0.7  1040.8 ± 87.7  4 2 . 5 - 4.7  4 9 . 6 ± 3.5  5.2 - 0.6  850.2 ± 8 2 . 0  42.1 ± 6.9  52.1 - 6.4  s i g n i f i c a n t r e d u c t i o n i n l o n g e v i t y a t 100 ppm d i f l u b e n z u r o n .  cages/treatment.  3.  Seasonal abundance o f f o u r l e a f r o l l e r s p e c i e s i n the t e s t o r c h a r d d u r i n g summer, 1980.  A A A • • • • ••  C rosaceana f\ limitata rosanus  Table XVIII.  Percent o f harvested apples showing l e a f r o l l e r  damage.  Treatment  94 ppm diflubenzuron  375 ppm diflubenzuron  375 ppm azinphos-methyl  A  -  -  -  ' -  B  +  -  -  -  C  +  -  -  +  +•  -  -  . 7;9 ± 0.9  -  +  -  1.9 ± 0.3  D E  ' Contrasts:  (A) (B) (B) (D)  vs. vs. vs. vs.  500 ppm Tween 20  % damage (mean - S . E . ) 15.8 ± 1.4 12.1 -  •  (B+C+D+E) - h i g h l y s i g n i f i c a n t . (C) - not s i g n i f i c a n t . (D) - s i g n i f i c a n t . (E) - ^ s i g n i f i c a n t .  0.9  10.2 ± 0.8  -48-  l e a f r o l l e r damage i n the t e s t o r c h a r d .  The high number o f a d u l t s  present  i s r e f l e c t e d i n a high damage r a t e i n the check t r e a t m e n t , where 15.8% f r u i t damage o c c u r r e d .  At recommended r a t e s (375 ppm) a z i n p h o s - m e t h y l was the ,  most e f f e c t i v e t r e a t m e n t t e s t e d , as o n l y 1.9% i n j u r y r e s u l t e d .  A t compar-  a b l e r a t e s , d i f l u b e n z u r o n reduced the i n f e s t a t i o n , but c o n s i d e r a b l y than a z i n p h o s - m e t h y l .  The lower-  less  d i f l u b e n z u r o n c o n c e n t r a t i o n was l e s s  effec-  t i v e and p e r c e n t damage was not s i g n i f i c a n t l y reduced by the a d d i t i o n o f Tween 2 0 . B >  C o d l i n g Moth T e s t s F i g . 4 shows the seasonal abundance o f the c o d l i n g moth i n o r c h a r d s  a t the Summerland Research S t a t i o n d u r i n g t h e summer o f 1980. were observed t o emerge, the f i r s t in e a r l y August.  i n l a t e May to e a r l y June and the second  Large numbers o f f i r s t - g e n e r a t i o n moths emerged i n m i d -  May, a p p r o x i m a t e l y a t the p e t a l - f a l l sufficiently  Two broods  stage.  F r u i t would not have developed  a t t h i s stage t o p r o v i d e food f o r young l a r v a e .  It is  possible  t h a t the m i l d w i n t e r and hot s p r i n g o f 1979-1980 induced premature emergence of the f i r s t  brood.  Poor s u r v i v a l  o f t h i s f i r s t g e n e r a t i o n may have been  the major reason f o r the low subsequent  infestations.  C o d l i n g moth damage t o h a r v e s t e d a p p l e s from the m i x e d - c u l t i v a r i s shown i n T a b l e XIX.  A check t r e a t m e n t was not i n c l u d e d , but  o f b u f f e r t r e e s gave an approximate v a l u e o f 2.0% damage. is s t i l l  above the economic t h r e s h o l d o f c a . 0.5-1.0%.  v e r y few f r u i t ,  so were not i n c l u d e d i n the a n a l y s i s .  to g i v e b e t t e r c o n t r o l  orchard  examination  While l o w ,  The Rome t r e e s  this set  D i f l u b e n z u r o n appeared  than a z i n p h o s - m e t h y l when a p p l i e d a t recommended r a t e s  (187 ppm), but the two t r e a t m e n t s were not s i g n i f i c a n t l y d i f f e r e n t .  The  presence o f Tween 20 s i g n i f i c a n t l y enhanced the a c t i o n o f 47 ppm d i f l u b e n -  F i g . 4.  Seasonal abundance o f the c o d l i n g moth in*Summerland o r c h a r d s summer, 1980. S p r a y . d a t e s are i n d i c a t e d by a r r o w s .  during  T a b l e XIX.  Treatment  Percent o f harvested apples o f mixed c u l t i v a r s  showing c o d l i n g moth damage.  47 ppm diflubenzuron  187 ppm diflubenzuron  187 ppm azinphos-methyl  A  +  -  -  B  +  -  C  -  +  D  1  —  Contrasts:  e s t i m a t e d check i n f e s t a t i o n = 2.0%.  500 ppm Tween 20  % damage (mean - S . E . ) 2.0  +  0.8  +  +  0.7 0.1  •-  0.4  +  0.1  +  0.7  +  0.2  (A) v s . (B) - s i g n i f i c a n t . (A) v s . (C) - s i g n i f i c a n t . .(C) v s . (D) - not s i g n i f i c a n t .  zuron.  The h i g h e r d i f l u b e n z u r o n c o n c e n t r a t i o n gave s i g n i f i c a n t l y  control  than t h e lower d i f l u b e n z u r o n  better  concentration.  T a b l e XX shows the c o d l i n g moth damage to h a r v e s t e d Golden D e l i c i o u s apples.  The i n f e s t a t i o n i n the check t r e a t m e n t was s i g n i f i c a n t l y  than t h e average o f the o t h e r f o u r t r e a t m e n t s , but no f u r t h e r  greater  differences  between t r e a t m e n t s were f o u n d . C.  Mites The use o f c o d l i n g moth c o n t r o l measures which do not d i s r u p t  mites i s c r i t i c a l  i n a p p l e pest management.  Two f a c t o r s which a r e  predaceous critical  i n d e t e r m i n i n g European red m i t e i n f e s t a t i o n s a r e the numbers o f r e d m i t e s per l e a f  (economic t h r e s h o l d c a . 15-30) and numbers o f red m i t e s per p h y t o -  s e i i d m i t e (economic t h r e s h o l d c a . 10:1)  (Downing and A r r a n d 1978).  t h r e s h o l d s s h o u l d be exceeded b e f o r e p r o c e e d i n g w i t h a c a r i c i d a l  Both  sprays.  As  shown i n F i g . 5, none o f the t r e a t m e n t s used a g a i n s t the c o d l i n g moth r e s u l t e d i n dangerous l e v e l s o f red m i t e s .  The most common p r e d a t o r was t h e  p h y t o s e i i d Typhlodromus o c c i d e n t a l i s N e s b i t t , but o t h e r p h y t o s e i i d and the s t i g m a e i d Z e t z e l l i a m a l i  (Ewing) were p r e s e n t as w e l l .  e x a m i n a t i o n found no e v i d e n c e o f t o x i c i t y t o t h e s e p r e d a t o r s .  species  Microscopic While r u s t  m i t e s were p r e s e n t throughout the s e a s o n , they were not abundant and never exceeded 10 mites per l e a f .  The McDaniel s p i d e r m i t e (Tetranychus  mcdanieli  McGregor) and t h e t w o - s p o t t e d s p i d e r m i t e (J_. u r t i c a e (Koch)) were seldom observed.  T a b l e XX.  Percent o f h a r v e s t e d Golden D e l i c i o u s apples showing c o d l i n g moth damage.  Treatment  47 ppm diflubenzuron  187 ppm diflubenzuron  187 ppm azinphos-methyl  '.A  -  -  -  2.1  B  +  -  -  0.8  C  +  -  D  -  +  '  +  E Contrasts:  500 ppm Tween 20  (A) (B) (B) (D)  vs. vs. vs. vs.  % damage (mean ±*S.E.)  0.8  -  0.5  +  0.4  (B+C+D+E) - s i g n i f i c a n t . (C) - not s i g n i f i c a n t . (D) - not s i g n i f i c a n t . (E) - not s i g n i f i c a n t .  +  +  +  +  +  0.2 0.1 0.1 0.1 0.1  -53-  F i g . 5.  10  J  European red m i t e numbers, per l e a f and per p h y t o s e i i d , i n t h e c o d l i n g moth f i e l d t e s t o r c h a r d s d u r i n g summer, 19,80. Spray dates are i n d i c a t e d by a r r o w s . :  DISCUSSION D u r i n g the l a s t two decades, a p p l e i n s e c t c o n t r o l e v o l v e d away from a t o t a l  r e l i a n c e on i n s e c t i c i d e s  By-incorporating biological  and c u l t u r a l  strategies  (Hoyt and B u r t s  1974).  c o n t r o l s wherever f e a s i b l e ,  w i t h c a r e f u l a p p l i c a t i o n o f more s e l e c t i v e p e s t i c i d e s , chemical measures can be s i g n i f i c a n t l y r e d u c e d .  have  along  control  Pheromone t r a p s and o t h e r  monitoring  t e c h n i q u e s can be used to determine numbers o f pest i n s e c t s and dates peak a c t i v i t y .  T h i s c o n t r i b u t e s t o a r e d u c t i o n i n chemical s p r a y s by a l -  l o w i n g g r e a t e r e f f i c i e n c y and a c c u r a c y i n a p p l i c a t i o n o f p e s t i c i d e s e t al_. 1 9 7 4 ) .  of  Sterile  (Myburgh  i n s e c t r e l e a s e and the pheromone c o n f u s i o n method  show promise and may make v a l u a b l e c o n t r i b u t i o n s to f u t u r e apple p e s t management (Hoyt and B u r t s 1974).  I t seems l i k e l y ,  however, t h a t p e s t i c i d e s  remain the major r e c o u r s e o f the p e s t manager i n the f o r e s e e a b l e Organophosphate i n s e c t i c i d e s t i v e l y f o r control of apple p e s t s . o f many b e n e f i c i a l (Hoyt 1969).  will  future.  such as a z i n p h o s - m e t h y l are used e f f e c T h i s i s due i n p a r t to the  resistance  a r t h r o p o d s , n o t a b l y predaceous m i t e s , to the compounds  Until  t h i s r e s i s t a n c e d e v e l o p s , a p e s t i c i d e may be e x t r e m e l y  n o n s e l e c t i v e and e l i m i n a t e n a t u r a l bamate i n s e c t i c i d e s ,  enemies o f a number o f p e s t s .  The c a r -  f o r example, a r e not as s e l e c t i v e as t h e organophos-  phates (Downing and A r r a n d 1 9 7 8 ) , p r o b a b l y s i n c e the l a t t e r have been i n use f o r much l o n g e r p e r i o d s .  However, c o n c u r r e n t r e s i s t a n c e t o organophosphates  has a l s o been observed w i t h a number o f l e s s e r a p p l e p e s t s i n c l u d i n g rollers,  l e a f h o p p e r s , aphids and l e a f miners ( C r o f t 1 9 7 9 ) .  Therefore,  leafit  is  p o s s i b l e t h a t r e s i s t a n c e may develop i n major p e s t s such as the c o d l i n g moth, p a r t i c u l a r l y i n areas w i t h i n the U n i t e d S t a t e s where growers may have been  -55-  u s i n g organophosphorous compounds f o r over 20 y e a r s  (Croft  1979).  I t i s p o s s i b l e t h a t d i f f e r e n t groups of c h e m i c a l s may be r e q u i r e d f o r c o n t r o l o f a p p l e p e s t s i n the near f u t u r e .  Such a change i n c o n t r o l  measures might r e s u l t i n a d i s r u p t i o n o f b e n e f i c i a l broad-spectrum e f f e c t s .  Until  a r t h r o p o d s because o f  r e s i s t a n t s t r a i n s of these species  a p p l e p e s t management would be much l e s s e f f i c i e n t than i t  i s at  develop, present.  The carbamates and p a r t i c u l a r l y the s y n t h e t i c p y r e t h r o i d s a r e c a n d i d a t e s f o r a p p l e pest c o n t r o l  (Hoyt e t al_. 1978).  likely  The p y r e t h r o i d s  o f low mammalian t o x i c i t y and are h i g h l y t o x i c to many p e s t s p e c i e s and Hoyt 1978).  However, the problem o f n o n s p e c i f i c i t y ,  are  (Croft  particularly  against  predaceous m i t e s , has a l r e a d y been observed (Hoyt e_t al_. 1978; Wong and Chapman 1979). As d e s c r i b e d i n the I n t r o d u c t i o n , f i e l d s t u d i e s have shown t h a t d i f l u benzuron has c o n s i d e r a b l e p o t e n t i a l  i n the i n t e g r a t e d c o n t r o l o f o r c h a r d  p e s t s (Cranham 1978; Wearing and Thomas 1978; W e s t i g a r d 1979). has a low mammalian t o x i c i t y  The compound  (Anonymous 1974) and would pose l e s s  t o the a p p l i c a t o r than most r e g i s t e r e d compounds. i s more s e l e c t i v e than many c o n v e n t i o n a l  hazards  In a d d i t i o n , the compound  insecticides.  Because d i f l u b e n z u r o n  i s n o n - s y s t e m i c i n p l a n t s ( B u l l and I v i e 1978) and a c t s p r i m a r i l y as a stomach p o i s o n (Post and V i n c e n t 1 9 7 3 ) , i t  i s t o x i c to p l a n t - f e e d i n g  but not d i r e c t l y t o x i c t o many p r e d a t o r y and p a r a s i t i c a r t h r o p o d s mous 1974).  P r e d a t o r y m i t e s a r e o f p a r t i c u l a r importance i n a p p l e  (Anonyprotec-  t i o n and s t u d i e s i n New Zealand (Wearing and Thomas 1 9 7 8 ) , A u s t r a l i a and K a l d o r 1980) and the U.S.  (Westigard 1979) suggest t h a t  i s n o n - t o x i c to predaceous m i t e s .  species  (Bower  diflubenzuron  As d i f l u b e n z u r o n i s a l s o o f low t o x i c i t y  t o honeybees (Anonymous 1974), use o f the compound i n a p p l e p e s t management  would l i k e l y  have few a d v e r s e s i d e - e f f e c t s  against beneficial  organisms.-  As m e n t i o n e d , a number o f f i e l d s t u d i e s have i n d i c a t e d t h a t zuron e f f e c t i v e l y c o n t r o l s t h e c o d l i n g moth.  However, t h e s e t e s t s d i d not  e s t a b l i s h which l i f e s t a g e s were most s e n s i t i v e .  When c h o o s i n g the d a t e ,  f o r m u l a t i o n and method o f a p p l i c a t i o n o f s p r a y s , s e n s i t i v e l i f e s h o u l d be c o n s i d e r e d .  The c u r r e n t s t u d i e s  difluben-  stages  i n d i c a t e t h a t t h e eggs and young  l a r v a e o f the c o d l i n g moth a r e t h e o n l y stages s u s c e p t i b l e to d i f l u b e n z u r o n . A d u l t moths showed no marked a d v e r s e e f f e c t s when dipped i n benzuron s o l u t i o n s , a l t h o u g h m a r g i n a l r e d u c t i o n s b i l i t y were o b s e r v e d .  or ' f e d  diflu-  i n f e c u n d i t y and egg v i a -  S i n c e t h e t r e a t m e n t s were l i k e l y more severe than  s p r a y s which moths would encounter i n an o r c h a r d s i t u a t i o n , c o n t r o l o f a d u l t moths seems most u n l i k e l y .  The a p p a r e n t immunity o f a d u l t s  d i f l u b e n z u r o n a t a d i s a d v a n t a g e when compared t o customary  the  puts  insecticides.  A z i n p h o s - m e t h y l , f o r example, i s t o x i c to eggs ( W e l l s and Guyer 1 9 6 2 ) , l a r v a e ( G r a t w i c k e t al_. 1965) and a d u l t s  ( M o f f i t t and White 1972) o f  the  c o d l i n g moth. T o x i c i t y o f d i f l u b e n z u r o n to eggs and l a r v a e o f . t h e c o d l i n g moth must t h e r e f o r e be e x t r e m e l y h i g h i n o r d e r t o match t h e e f f i c a c y o f compounds.  The eggs appear v e r y s e n s i t i v e , e s p e c i a l l y when t r e a t e d  shortly after oviposition. trol  conventional topically  One hundred ppm d i f l u b e n z u r o n p r o v i d e d 95% c o n -  o f 0- t o 2 5 - d a y - o l d eggs, and younger eggs were even more s e n s i t i v e .  The r e s i d u a l  ovicidal  a c t i v i t y o f d i f l u b e n z u r o n was e x t r e m e l y good when  d r i e d on f r u i t and f o l i a g e and 'did not n o t i c e a b l y d i m i n i s h 10 days  after  application.  When t r e a t e d f o l i a g e was sprayed w i t h water on a d a i l y  the o v i c i d a l  a c t i o n was a p p a r e n t l y r e d u c e d , but not t o a s i g n i f i c a n t  C o n s e q u e n t l y , extended c o n t r o l  o f eggs would be expected because o f  it  basis, extent. the  p e r s i s t e n c e o f d i f l u b e n z u r o n on f r u i t and f o l i a g e . The use o f Tween 20 was e s s e n t i a l  in laboratory tests for  improving  r e t e n t i o n and d i s t r i b u t i o n o f t e s t s o l u t i o n s on s u b s t r a t e s .  In the absence  of the s u r f a c t a n t ,  particularly  s o l u t i o n s tended to bead up and drop o f f ,  on waxy mature a p p l e s .  Immature, a p p l e s and f o l i a g e showed b e t t e r  o f aqueous s o l u t i o n s , p a r t i c u l a r l y when Tween 20 was added.  retention  When e v a l u a t e d  i n f i e l d t e s t s , however, the s u r f a c t a n t d i d not appear to improve t h e i n s e e ticidal  action of diflubenzuron.  While one c o d l i n g moth f i e l d t e s t  (mixed  c u l t i v a r o r c h a r d ) d i d suggest such an improvement, v a r i a b l e and low i n f e s t a t i o n l e v e l s c a s t doubt on t h i s .  F u r t h e r m o r e , i n the  heavily-infested  l e a f r o l l e r o r c h a r d , no s i g n i f i c a n t enhancement was o b s e r v e d . many reasons why Tween 20 d i d not perform w e l l  There may be  i n the f i e l d ,  but the  emphasizes the importance o f f i e l d t e s t i n g as a f o l l o w - u p to  result  laboratory  studies. The mode o f a c t i o n o f d i f l u b e n z u r o n as an o v i c i d e i s not w e l l stood.  under-  Younger c o d l i n g moth eggs were more s e n s i t i v e t o the compound than  o l d e r eggs; t h i s was a l s o noted f o r Spodoptera 1 i t t o r a l i s Ascher ejt aJL  (1980).  ( N o c t u i d a e ) by  A t the same t i m e , s e n s i t i v i t y was g r e a t e r i f  the  d i f l u b e n z u r o n remained i n s o l u t i o n on c o d l i n g moth eggs as opposed t o d r y i n g rapidly.  The presence o f Tween 20 i n t h e t e s t s o l u t i o n s a l s o  sensitivity.  increased  I t i s p o s s i b l e t h a t d i f l u b e n z u r o n must e n t e r the eggs w h i l e  s o l u t i o n , s i n c e the compound i s not v o l a t i l e  ( M e t c a l f e_t aj_. 1 9 7 5 ) .  in  Entry  may t a k e p l a c e through the a e r o p y l e s and/or c h o r i o n , i n which c a s e the r e duced s u r f a c e t e n s i o n e f f e c t e d by Tween 20 might f a c i l i t a t e uptake.  However, i t must be remembered t h a t the r e s i d u a l  diflubenzuron  ovicidal  action  o f d i f l u b e n z u r o n was high when the compound had d r i e d on f r u i t and f o l i a g e .  Eggs would c o n t a c t d i f l u b e n z u r o n a t o v i p o s i t i o n and would c e r t a i n l y be m o i s t w i t h s e c r e t i o n s from the a c c e s s o r y g l a n d s , which cement the eggs to o v i p o s i tion sites.  T h i s m o i s t u r e might be s u f f i c i e n t to m o b i l i z e the  a g a i n s t the h i g h l y s u s c e p t i b l e young eggs.  diflubenzuron  Ascher et_ al_. (1980) observed  t h a t a l i q u i d f o r m u l a t i o n o f d i f l u b e n z u r o n was about ten times more a c t i v e a g a i n s t S_. l i t t o r a l i s eggs than was the w e t t a b l e powder f o r m u l a t i o n .  Eggs  ° f L o b e s i a botrana ( T o r t r i c i d a e ) were more s e n s i t i v e to d i f l u b e n z u r o n  at  h i g h e r temperatures  that  (Ascher e_t aj_. 1978).  It is possible, therefore,  under c e r t a i n c o n d i t i o n s c o d l i n g moth eggs may be even more s e n s i t i v e d i f l u b e n z u r o n than r e v e a l e d i n the c u r r e n t s t u d y , s i n c e a l l  to  t e s t s were p e r -  formed a t room temperature ( c a . 25°C) w i t h a w e t t a b l e powder f o r m u l a t i o n . Most d i f l u b e n z u r o n - t r e a t e d c o d l i n g moth eggs developed t o the p o i n t o f e c l o s i o n ( t h e " b l a c k h e a d " stage) but the l a r v a e were unable to break the c h o r i o n and emerge.  I f the c h o r i o n s were opened and the l a r v a e a i d e d t o  emerge, they remained l e t h a r g i c , d i d not f e e d , and i n v a r i a b l y d i e d . c u t i c l e s appeared much more d e l i c a t e than those o f h e a l t h y l a r v a e , e a s i l y and causing- d e s i c c a t i o n .  Since t h i s condition represents  symptoms o f d i f l u b e n z u r o n p o i s o n i n g , i t appears t h a t the compound c h i t i n synthesis  i n the pharate c u t i c l e o f d e v e l o p i n g embryos  i f absorbed d u r i n g e a r l y  Their rupturing  classic inhibits  particularly  embryogenesis.  L a b o r a t o r y t e s t s showed t h a t d i f l u b e n z u r o n was a l s o t o x i c to young c o d l i n g moth l a r v a e .  I f s u f f i c i e n t d i f l u b e n z u r o n was i n g e s t e d , f i r s t -  2 n d - i n s t a r l a r v a e proved i n c a p a b l e o f m o u l t i n g t o subsequent i n s t a r s .  and Since  no t o x i c e f f e c t s were observed except d u r i n g the m o u l t , f r u i t e n t r y may occur b e f o r e d e a t h , l a v i n g a small b l e m i s h . r e a d i l y on d i f l u b e n z u r o n - t r e a t e d f o l i a g e ,  As f i r s t - i n s t a r  l a r v a e fed  i t appears l i k e l y t h a t  significant  -59-.  l a r v a l m o r t a l i t y would o c c u r i n the  field.  In f i e l d t e s t s a t Summerland, d i f l u b e n z u r o n appeared equal t o a z i n p h o s methyl i n s u p p r e s s i n g c o d l i n g moth damage.  U n f o r t u n a t e l y , the c o d l i n g moth  i n f e s t a t i o n was low i n t h e t e s t o r c h a r d s and d i f f e r e n c e s between t r e a t m e n t s were d i f f i c u l t  to d e t e c t .  In a h e a v i l y - i n f e s t e d o r c h a r d , d i f l u b e n z u r o n was  not as e f f e c t i v e as a z i n p h o s - m e t h y l K a l d o r 1980).  f o r c o d l i n g moth c o n t r o l  However, i n t h i s s t u d y sprays were not s y n c h r o n i z e d w i t h  peak a d u l t a c t i v i t y .  As demonstrated i n the c u r r e n t s t u d y , f r e s h l y  p o s i t e d eggs a r e the most s e n s i t i v e s t a g e so c a r e f u l the use o f d i f l u b e n z u r o n . o f a d u l t moths.  (Bower and  timing is essential  Sprays should be a p p l i e d s h o r t l y a f t e r  Other f i e l d t e s t s  sensitive  These masses are covered w i t h a g e l a -  t i n o u s c e m e n t - l i k e s e c r e t i o n from the a c c e s s o r y glands and t h i s This i n s e n s i t i v i t y  substance  i s p u z z l i n g i n view o f  the f a c t t h a t S_. l i t t o r a l i s eggs a r e l a i d i n masses but are s t i l l t i b l e to d i f l u b e n z u r o n (Ascher e t aJL 1980).  I t appears t h a t f i r s t - i n s t a r  a r e s e n s i t i v e t o d i f l u b e n z u r o n when i t  suscep-  L e a f r o l l e r a d u l t s showed a  s l i g h t r e d u c t i o n i n l o n g e v i t y when dipped i n 100 ppm d i f l u b e n z u r o n but no o t h e r a d v e r s e e f f e c t s .  emergence  control.  Egg masses o f the obiiquebanded l e a f r o l l e r d i d not appear  may p r e v e n t a c c e s s to the e g g s .  in  (Wearing and Thomas 1978; Cranham 1978;  W e s t i g a r d 1979) have r e s u l t e d i n e x c e l l e n t  to d i f l u b e n z u r o n even a t 1,000 ppm.  ovi-  solutions,  leafroller  i s i n g e s t e d from f o l i a g e .  larvae  Other  in-  s t a r s may be s u s c e p t i b l e as w e l l , but t h i s c o u l d not be c o n c l u s i v e l y demonstrated.  The l a b o r a t o r y s t u d i e s , t h e r e f o r e , d i d not i n d i c a t e good p o t e n t i a l  f o r d i f l u b e n z u r o n as a c o n t r o l agent f o r l e a f r o l l e r s . leafrollers  r e f l e c t e d the l a b o r a t o r y f i n d i n g s .  Field tests  against  While d i f l u b e n z u r o n d i d  n i f i c a n t l y reduce l e a f r o l l e r damage, a z i n p h o s - m e t h y l  gave c a . f o u r times  sig-  -60-  better control.than diflubenzuron. • Increases  i n numbers o f a p p l e p e s t s might come about i n the f u t u r e  because o f 1) a r e d u c t i o n i n c o d l i n g moth and o t h e r sprays due to  sterility-  c o n t r o l o r i n t e g r a t e d pest management o r 2) t h e use o f n o n - s p e c i f i c  pesti-  c i d e s which d i s r u p t p r e d a t o r y and p a r a s i t i c a r t h r o p o d s .  likely  I t appears  t h a t d i f l u b e n z u r o n would not cause t h i s l a t t e r phenomenon and t h e r e f o r e  is  a s u i t a b l e candidate f o r i n v e s t i g a t i o n regarding control of apple p e s t s . L e a f r o l l e r s are not e f f e c t i v e l y c o n t r o l l e d by d i f l u b e n z u r o n , but the compound has c o n s i d e r a b l e p o t e n t i a l  f o r c o d l i n g moth c o n t r o l .  Since d i f l u b e n -  zuron i s e f f e c t i v e a g a i n s t immature stages but not a d u l t moths, i t s use i n conjunction with s t e r i l i t y - c o n t r o l compounds.  would be l e s s d i s r u p t i v e than c o n v e n t i o n a l  W e s t i g a r d (1979) showed t h a t d i f l u b e n z u r o n was l e s s t o x i c  n a t u r a l enemies o f the pear p s y l l a than were a z i n p h o s - m e t h y l I t might be o f v a l u e to t e s t o t h e r pome f r u i t , p e s t s  to  and phosmet.  f o r s e n s i t i v i t y to d i f l u -  b e n z u r o n , g i v e n t h e l i k e l i h o o d t h a t use o f t h e growth r e g u l a t o r would not greatly disrupt beneficial  organisms.  -61-  LITERATURE CITED A b b o t t , W.S. ticide.  1925. 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E n t . 69: 365-367.  -68-  APPENDIX Table la.  P e r c e n t hatch o f 3 - d a y - o l d c o d l i n g moth eggs t o p i c a l l y with diflubenzuron s o l u t i o n s . 50 eggs per r e p l i c a t e .  Replicate  0 ppm  1 2 3 4  84 76 78 80  70 76 66 64  46 62 38 44  20 16 16 22  X  79.5  69.0  47. 5  18.5  A n a l y s i s of  1 ppm  10 ppm  100 ppm  variance  Source  DF  S_S  MS  Total Treatment Linear regression Quadratic regression Deviations Error  15 3 1 1 1 12  9169. 8 8708. 8 7214. 1 1387. 4 no. 0 461. 0  2902.9 7214.1 1387.4 110.0 38.4  Table . lb. Replicate  treated  75.56* 187.78* 36.11* 2.86  P e r c e n t hatch o f 0- to 2 j - d a y - o l d c o d l i n g moth eggs t o p i c a l l y t r e a t e d w i t h d i f l u b e n z u r o n s o l u t i o n s . 50 eggs per r e p l i c a t e . 0 ppm  0;.01 ppm  0,.1 ppm  10 .0 ppm  1.,0 ppm  100 ppm  1 2 3 4 5 6 7 8 9  78 58 64 72 '58 72 68 70 78  64 78 56 78 76 72 68 74 68  54 50 52 40 44 66 52 46 46  38 46 42 44 46 34 36 40 28  8 0 0 18 14 12 8 6 14  12 4 6 4 2 6 0 4 6  X  68.7  70.4  50.0  39.3  8.9 .  4.9  Analysis of  variance  ( e x c l u d i n g 0 ppm from a n a l y s i s )  Source  DF  Total Treatment log Linear regression log Quadratic regression l o g Cubic r e g r e s s i o n log Deviations Error  44 4 1 1 1 1 40  SS 29280.1 27788.1 26694.4 103.1 250.0 738.3 1491.1  MS V6947; 2 26694. 4 103. 1 250. 0 738. 3 37. 3  F 186.37 * 716.11 * 2.77 6.71 * 19.80 *  -69-  Table Ilia.  P e r c e n t hatch o f c o d l i n g moth eggs t o p i c a l l y t r e a t e d w i t h 10 ppm d i f l u b e n z u r o n a t d i f f e r e n t l e n g t h s o f time from o v i p o s i t i o n . 50 eggs per r e p l i c a t e .  Replicate  0 - 36 h  36 - 60 h  60 - 84 h  84 - 108 h  108 - 132  1 2 3 4 5 6 7 8 9  12 26 32 8 10 22 10 12 8  32 14 18 18 22 18 20 8 12  52 36 64 40 42 28 44 36 30  48 30 34 36 64 40 40 32 34  46 70 56 58 62 72 64 66 56  X  15.6  18.0  41.3  39.8  61.1  A n a l y s i s o f 'v a r i a n c e Source  DF  Total Treatment Linear regression Quadratic regression Cubic r e g r e s s i o n Deviations Error  44 4 •1 1 1 1 40  Table 111 b.  MS_  SS 16108.0 12682.6 11925.0 146.8 5.4 1349.3 3425.4  F_  3170.7 11925.0 146.8 5.4 1349.3 85.6  37.03* 139.26* 1.71 0.06 15.76*  P e r c e n t hatch o f young c o d l i n g moth eggs t o p i c a l l y t r e a t e d w i t h 10 ppm d i f l u b e n z u r o n a t d i f f e r e n t l e n g t h s of time from o v i p o s i t i o n . 10 eggs per r e p l i c a t e  )licate 1 2 3 4 X A n a l y s i s of  0 - 12 h 0 . 10 0 10 5.0  12 - 24 h  24 - 36 h  36 - 48 h  40 50 30 40  20 60 20 30  40 80 50 70  40.0  32.5  60.0  variance  Source  DF_  SS  MS  F  Total Treatment Linear regression Quadratic regression Deviations Error  15 3 1 11 1 12  8969 .8 6218 .8 4961 .3 56 .3 1201 .2 2551 .0  2072.9 4961.3 56.3 1201.2 212.6  9.75* 23.31* 0.26 5.65*  -70-  Table IVa.  .  P e r c e n t hatch o f 3 - d a y - o l d c o d l i n g moth eggs p l a c e d i n c o n t a c t w i t h 10 ppm d i f l u b e n z u r o n s o l u t i o n f o r v a r y i n g p e r i o d s o f t i m e , d i f l u benzuron a p p l i e d w i t h a f i n e b r u s h . 50 eggs per r e p l i c a t e . 2.0 h  4.0 h  Replicate  0.25 h  0.,5 h  1.0 h  1 2 3 4  74 80 78 78  70 72 66 70  74 74 70 68  62 64 62 58  32 46 44 40  X  77.5  69.5  71.5  61.5  40.5  Analysis  of v a r i a n c e  Source  DF  SS  MS  F  Total Treatment log Linear regression log Quadratic regression l o g Cubic r e g r e s s i o n Tori D e v i a t i o n s Error  19 4 1 1 1 1 15  3503.6 3308.8 2689.6 412.6 176.4 30.2 194.8  827.2 2689.6 412.6 176.4 30.2 13.0  63.68* 207.05* 31.76* 13.58* 2.33  Table IVb.  P e r c e n t hatch o f 0 - t o 2 £ - d a y - o l d c o d l i n g moth eggs p l a c e d i n c o n t a c t w i t h 10 ppm d i f l u b e n z u r o n s o l u t i o n f o r v a r y i n g p e r i o d s o f t i m e , d i f l u b e n z u r o n a p p l i e d i n soaked paper t o w e l l i n g . 50 eggs per replicate.  icate  0.5 h  1.0 h  2.0 h  60 62 58 68 66 56 52 48 72  4.0 h  64 60 58 52 58 60 64 62 48  34 54 34 36 48 30 38 22 34  58.4  36.7  1 2 3 4 5 6 7 8  80 72 62 58 74 88 76.. 86 88  64 70 62 74 70 58 52 54 74  X  76.0  64.2  . 60.2  Source  DF  s_s  MS  F_  Total Treatment log Linear regression log Quadratic regression l o g Cubic r e g r e s s i o n log Deviations Error  44 4 1 1 1 1 40  8469 .3 7457 .5 6417 .8 203 .2 694 .4 35 .7 1011 .8  1864 .4 6417 .8 203 .2 694 .4 35 .7 25 .3  73.69* 253.67* 8.03* 27.45* 1.41  9-  A  0.25 h  A n a l y s i s of  variance  -71-  Table Va.  P e r c e n t hatch o f 3 - d a y - o l d c o d l i n g moth eggs t r e a t e d t o p i c a l l y w i t h 10 ppm d i f l u b e n z u r o n i n the presence and absence o f 500 ppm Tween 20. 50 eggs per r e p l i c a t e .  Replicate  (A) 500 ppm Tw20  (B) 10 ppm d i f  (C) 10 d i f + 500 Tw20  1 2 3 4  84 • 80 78 86  48 34 42 38  16 12 18 22  X  82.0  40.5  17.0  A n a l y s i s of  variance  Source  DF  SS  MS  Total Treatment (A+B) v s . (C) (A) v s . (B) Error  11 2 1 1 9  8865.0 8666.0 5221.5 3444.5 199.0  4333.0 5221.5 3444.5 22.1  Table Vb.  f_ 196.06* 236.27* 155.86*  P e r c e n t hatch o f 1- t o 2 - d a y - o l d c o d l i n g moth eggs t r e a t e d t o p i c a l l y w i t h d i f l u b e n z u r o n s o l u t i o n s i n the presence and absence o f 500 ppm Tween 20. 50 eggs per r e p l i c a t e .  Replicate  0 ppm d i f  0 ppm+Tw  1 ppm  1 ppm+Tw  10 ppm  10 ppm+Tw  1 2 3 4 5 6 7 8 9  68 74 82 74 82 76 76 72 80  80 68 82 70 84 74 78 74 78  80 64 76 66 56 72 70 62 74  78 68 70 60 60 54 58 64 58  38 50 48 52 46 32 36 28 30  42 28 40 38 16 42 34 28 26  X  76.0  76.4  68.9  63.3  40.0  I  32.7  Anal ys i s ^ o ; l v a r i ance :  Source  DF  Total Treatment Diflubenzuron Tween 20 D i f X Tw20 i n t e r a c t i o n Error  53 5 2 1 2 48  SS 18469 .3 15862 .2 15480 .4 232 .2 149 .6 2607 .1  MS  F  3172.4 7740.2 232.2 74.8 54.3  58.42* 142.55* 4.28* 1.38  -72-  Table Via.  r.  H a t c h a b i l i t y o f c o d l i n g moth eggs o v i p o s i t e d on mature S p a r t a n a p p l e s which had been dipped i n d i f l u b e n z u r o n s o l u t i o n s i n the presence and absence o f Tween 20. One a p p l e = one r e p l i c a t e .  Treatment  Rep.  eggs hatched  0 ppm d i f  1 2 3 4 5 6  63 114 71 141 58 86  78 144 90 172 76 97  80.8 79.2 78.9 82.0 76.3 88.7 -  1 2 3 4 5 6  79 82 68 95 97 57  107 109 83 118 137 82  73.8 75.2 81.9 80.5 70.8 69.5  1 2 500 ppm Tw20 3 4 (C) 5 6  86 80 72 86 116 71  121 114 103 111 163 91  71 .1 70.2 69.9 77.5 71.2 78.0  +  0 ppm Tw20 (A)  100 ppm d i f +  0 ppm Tw20 (B)  100 ppm d i f +  Analysis of  total  eggs  p e r c e n t hatch  X  81.0  75.3  73.0  variance  Source  DF_  Total Treatment (A) v s . (B+C). (B) v s . (C) Error  17 2 1 1 15  MS 664.2 203.6 186.7 16.0 460.6  101.8 186.7 16.0 30.7  F 3.32 6.08* 0.52  -73-  Table VIb.  H a t c h a b i l i t y o f c o d l i n g moth eggs o v i p o s i t e d on immature Golden D e l i c i o u s a p p l e s which had been dipped i n d i f l u b e n z u r o n s o l u t i o n s i n the presence and absence o f Tween 2 0 . The a p p l e s i n the f o u r t h t r e a t m e n t were t r e a t e d 10 days e a r l i e r than the o t h e r s . One a p p l e = one r e p l i c a t e . eqqs hatched  total  eggs  percent  Treatment  Rep.  0 ppm d i f  1 2 3 4 5 6 7 8 9 TO. 11 12  85 43 61 86 48 82 82 82 79 79 48 89  120 70 84 151 64 135 110 135 113 122 71 123  70.8  1 2 3 4 5 6 7 8 9 10 11 12  42 37 36 24 44 23 38 43 32 42 24 29  86 92 77 68 96 44 95 129 121 124 58 75  48.8  1 2 3 4 5 6 7 8 9 10 11 12  23 26 14 25 47 21 31 12 25?. 19 20 12  102 143 106 93 212 93 99 71 113 135 "'95 51  22.6 18.2 13.2 26.9 22.2 22.6 31.3 16.9 22.1 14.1 21.0 23.5  +  0 ppm Tw20 (A)  v  100 ppm d i f +  0 ppm Tw20 (B)  100 ppm d i f +  500 ppm Tw20 (C)  61.4 72.6 56.9 75.0 60.7 74.6 60.7 69.9 64.7 67.6 72.4  40.2 46.7 35.3 45.8 52.3 40.0 33.3 26.4 33.9 41 .4 38.7  67.3  40.2  21.2  -74-  Table Vlb  (concluded)  Treatment  Rep.  eggs hatched  1 2 3 4 5 6 7 8 9 10 11 12  13 12 16 19 37 27 18 13 10 14 22 13  100 ppm d i f +  500 ppm Tw20 ( a f t e r 10 days) (D)  Analysis of  eggs  p e r c e n t hatch  74 49 63 74 168 122 123 78 56 89 161 72  17.6' 24.5 25.4 25.7 22.0 22.1 14.6 16.7 17.9 15.7 13.7 18.1  X  19.5  variance  Source  DF  Total Treatment (A) v s . (B+C+D) (B) v s . (C+D) (C) v s . (D) Error  47 3 1 1 1 44  Table Vila.  total  SS 19297.6 17810.6 14639.0 3150.9 17.5 1487.0  MS  F  5936.9 14639.0 3150.9 17.5 33.8  175.68* 433.18* 93.24* 0.52  H a t c h a b i l i t y o f c o d l i n g moth eggs o v i p o s i t e d on pear f o l i a g e which had been dipped i n dif1ubenzuron/Tween 20 s o l u t i o n s . One l e a f = one r e p l i c a t e , l e a v e s not p l a c e d i n w a t e r .  Treatment 0 ppm d i f +  500 ppm Tw20  100 ppm d i f +  500 ppm Tw20  Rep.  eggs hatched  total  eggs  percent  1 2 3 4 5 6  155 63 86 26 58 45  200 84 107 29 77 55  77.5 75.0 80.4 89.7 75.3 81.8  1 2 3 4 5 6  10 25 20 27 32 18  23 105 61 59 98 47  43.5 23.8 32.8 45.8 32.6 38.3  X  79.9  36.1  -75-  Table V i l a (concluded). Analysis of Source  variance  .  Total Treatment Error Table Vllb.  DF  SS.  MS  11 1 10  6238.7 5755.3 483.4  5755.3 48.3  F  119.08*  H a t c h a b i l i t y o f c o d l i n g moth eggs o v i p o s i t e d on pear f o l i a g e which had been dipped i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s . The f o l i a g e i n the f o u r t h t r e a t m e n t was r e w e t t e d d a i l y w i t h d i s t i l l e d w a t e r . One l e a f = one r e p l i c a t e , w i t h the l e a v e s a t t a c h e d to t w i g s which were p l a c e d i n v i a l s o f w a t e r .  Treatment 0 ppm d i f • +  0 ppm Tw20 (A)  100 ppm d i f • •• • +  0 ppm Tw20 (B) ;  100 ppm d i f •+•  500 ppm Tw20 (C)  Rep.  eggs hatched  total  eggs  percent  1 2 3 4 5 6 7 8 9  29 35 36 22 34 40 33 27 36  29 36 36 22 41 41 39 28 40  100.0 97.2 100.0 100.0 82.9 97.6 84.6 96.4 90.0  1 2 3 4 5 6 7 8 9  22 44 16 3 12 12 6 7 3  22 52 25 16 34 45 14 21 11  100.0 84.6 64.0 18.8 35.3 26.7 42.9 33.3 27.3  1 2 3 4 5 6 7 8 9  4 20 9 3 9 2 5 5 6  21 40 21 13 29 30 16 29 31  19.1 50.0 42.9 23.1 31.0 6.7 31.3 17.2 19.4  X  94.3  48.1  26.7  -76-  Table v i ' l b (  c o n c  Treatment  "l ded) u  Rep.  100 ppm d i f + 500 ppm Tw20 (rewetted) (D)  A n a l y s i s of  eggs hatched  1 2 3, 4 5 6 7 8 9  10 11 23 7 3 5 10 7 2  eggs  p e r c e n t hatch  14 22 33 25 31 17 17 29 10  X  71.4 50.0: 69.7 28.0 9.7 29.4 58.8 24.1 20.0  40  variance  Source  DF  Total Treatment (A) v s . (B+C+D) (B) v s . (C+D) (C) v s . (D) Error  35 3 1 1 1 32  Table Xb.  total  MS_  SS 35656.5 23257.8 21168.0 1296.5 808.0 12398.7  f_  7752.6 21168.0 1296.5 808.0 387.5  20.01.* 54.63* 3.35 2.09  C o d l i n g moth f i r s t - i n s t a r l a r v a e e n t e r i n g a p p l e s and s u r v i v i n g to 4 t h - i n s t a r . Apples were d i p p e d i n dif1ubenzuron/Tween 20 solutions. 20 l a r v a e and a p p l e s per t r e a t m e n t Treatment  no. e n t e r i n g  .  no. t o  4th-instar  0 ppm d i f + 500 ppm Tw20  15  12  500 ppm d i f + 500 ppm Tw20  16  12  Table Xa,  C o d l i n g moth f i r s t - i n s t a r l a r v a e e n t e r i n g a p p l e s and s u r v i v i n g t o 4 t h - i n s t a r , a f t e r f e e d i n g on a p p l e f o l i a g e which had been sprayed w i t h d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s . Ten l a r v a e and a p p l e s per r e p l i c a t e .  Treatment 0 ppm d i f +  0 ppm Tw20 0 ppm d i f +  500 ppm Tw20 10 ppm d i f +  500 ppm Tw20 100 ppm d i f +  500 ppmTw20 500 ppm d i f +  500 ppm Tw20  Rep. 1 2 3 4  no. e n t e r i n g a p p l e s and feeding to 4 t h - i n s t a r a f t e r 1 day of f e e d i n g on f o l i a g e 9 e n t e r e d , 6 to 8,6 10,8 8,6  no. e n t e r i n g a f e e d i n g to 4th 2 days o f feed  4th-instar  8 entered, < 9,7 7,5 8,6  1 2 3 4  8,7 9,8 9,6 8,7  8,7 8,6 10,6 9,7  1 2 3 4  8,7 9,7 8,5 9,7  9,7 9,6 8,6 7,5  1 2 3 4  10,7 8,8 8,6 8,7  8,5 5,5 7,6 8,5  1 2 3 4  9,2 8,2 8,3 9,4  7,1 9,2 7,0 8,3  A n a l y s i s o f v a r i a n c e (on s u r v i v a l Source  DF  Total Treatment D i f l u b e n z u r o n (A) Days f e e d i n g (B) (A) X (B) i n t e r a c t i o n Error  39 9 4 1 4 30  to 4 t h - i n s t a r ) . SS 143.9 128.4 119.2 6.4 2.8 15.5  MS  f_  14.3 29.8 6.4 0.7 0.52  27.50* 57.31* 12.31* 1.36  -78-  Table Villa.  Replicate  M o r t a l i t y o f f i r s t - i n s t a r c o d l i n g moth l a r v a e p l a c e d on agar medium c o n t a i n i n g d i f f e r e n t c o n c e n t r a t i o n s o f d i f l u b e n z u r o n . F i v e l a r v a e per r e p l i c a t e , m o r t a l i t y assessed a f t e r 14 days o f f e e d i n g ( a f t e r Anderson 1978). 0 ppm d i f  10 ppm d i f  50 ppm d i f  100 ppm d i f  1 2 3 4  1 0 1 1  3 1 3 2  3 3 2 2  3 4 3 4  X  15%  45%  50%  70%  A n a l y s i s of  variance  Source  P£  SS  MS  F_  Total Treatment Linear regression Quadratic regression Deviations Error  15 3 1 1 1 12  8400 6200 4879 172 1149 1200  2067 4879 172 1149 100  20.67* 48.79* 1 .72 11.49*  Table VHIb.  Replicate  M o r t a l i t y o f 2 n d - i n s t a r c o d l i n g moth l a r v a e p l a c e d on agar medium containing d i f f e r e n t concentrations of diflubenzuron. Five l a r v a e per r e p l i c a t e , m o r t a l i t y a s s e s s e d a f t e r 10 days o f f e e d i n g ( a f t e r Anderson 1978). 0 ppm d i f  10 ppm d i f  50 ppm d i f  100 ppm d i f  1 2 3 4  1 0 1 2  3 4 4 2  4 5 4 4  5 5 5 4  X  20%  65%  85%  955?  A n a l y s i s of  t  variance  Source  DF  SS  Total Treatment Linear regression Quadratic regression Deviations Error  15. 3 1 1 1 12  15775 13275 9316 2117 1842 2500  MS 4425 9316 2117 1842 208 i  F 21.24* 44.72* 10.16* 8.84*  -7-9-  Table XI.  Treatment  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t c o d l i n g moths dipped i n d i f l u b e n z u r o n s o l u t i o n s . Four males and f o u r females per cage. eggs total percent Cage Sex L o n g e v i t y (days) hatched eggs hatch X  pm d i f  1 2 3  ppm d i f  1 2 3  ppm d i f  1 2 3  19 19 18 14 17 9  463  677  68.4  467  586  79.7  154  311  49.5  M F M F M F  3 , 14, 15, 16 2 , 3 , 13, 16 3 , 5, 7, 8 3, 9 , 14, 20 1 , 9 , 12, 14 1 , 3, 1 1 , 14  187  340  55.0  223  259  86.1  98  146  67.1  M F M F M F  3, 2, 8, 1, 3, 2,  81  167  48.5  149  211  70.6  162  206  78.6  M F M F M F  14 , 16, 4,13, 15 , 15, 8 , 13, 8 , io, 1 , 3,  3, 2, 9, 1, 7, 9,  18, 15, 17, 13, 16, 4,  11, 5, 11, 9, 12, 12,  14 12 17 13 13 14  65.9  69.4  65.9  A n a l y s i s of variance (fecundity) Source Total Treatment Linear regression Deviations Error  8 2 1 1 6  A n a l y s i s of variance (percent Source Total Treatment Linear regression Deviations Error  SS  MS  f_  280910.3 188135.1 90539.5 97600.0 92775.2  94067.6 90539.5 97600.0 15462.5  6.08* 5.86 6.31*  SS  MS  f_  1484.9 22.2 3.6 18.3 1462.7  11. 1 3. 6 18. 3 243. 8  DF  DF 8 2 1 1 6  hatch)  0.05 0.01 0.08  -80-  T a b l e XI  ,(concluded);  A n a l y s i s o f v a r i a n c e (sex X c o n c e n t r a t i o n , e f f e c t s on l o n g e v i t y ) Source  DF  Total Concentration Linear regression Deviations Cages/concentration Sex Sex X l i n e a r cone. Sex X d e v i a t i o n s cone. Sex X t o t a l cone. Sex X c a g e / c o n c e n t r a t i o n I n s e c t s / s e x and cage  71 (2) 1 1 6 1 1 1 (2) 6 54  Table XII. Treatment 0 ppm d i f  1 2 3  10 ppm d i f  1  +  0 ppm Tw20 (B)  2 3  100 ppm d i f  MS  2318 .4 289 .5 172 .2 117 .4 223 .4 156 .1 0 .3 100 .9 101 .2 133 .3 1414 .7  144 .8 172 .2 117 .4 37 .2 156 .1 0 .3 100 .9 50 .6 22 .2 26 .2  F_  3. 89 4. 62 3. 15 7. 02* 0. 01 4 . 54 2. 28  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t c o d l i n g moths dipped i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s . Four males and f o u r females per c a g e . eggs total percent Cage Sex L o n g e v i t y (days) hatched eggs hatch X  +  0 ppm Tw20 (A)  .  SS  1  +  0/ ppm' Tw20 (0  2  100 ppm d i f  1  3  +  500 ppm Tw20 2 (D) 3  M F M F M F  8, 6, 7, 5, 5, 2,  M F M F M F  5, 5, 6, 7, 2, 4,  M F M F M F M F M F M F  15, 13, 13, 12, 14, 8,  17 17 16 14 15 12  360  547  65.8  403  553  72.9  486  552  88.0  388  461  84.2  378  472  80.1  289  323  89.5  9 , 11, 12, 13 7, 8, 9, 14 5, 7, 8, 13 6, 8 , 9, 12 7, 10, 11» 16 8, 10, 13, 14  381  539  70.7  347  458  . 75.8  509  642  79.3  2, 5, 4 , 9, 5, 8 , 3, 9, 5, 7, 9, 10,  380  499  76.2  36  64  56.2  341  560  60.9  12, 11, 11. 6, 11» 6,  6, 8 , 14 7, 12, 16 8, 8 , 13 8, 11, 12 6, 12, 15 8, 9, 11  6, 10, 12, 13, 8, 12,  12 11 14 16 9 15  .  -81-  T a b l e 'XII  (concluded)  A n a l y s i s of  variance (fecundity)  Source  DF  SS  MS  Total Treatment (A) v s . (B+C+D) (B) v s . (C+D) (C) v s . (D) Error  11 3 1 1 1 8  249428.0 72298.1 24440.1 3472.2 44376.0 177129.9  24099.4 24440.1 3472.2 44376.0 22141.2  A n a l y s i s of  variance (percent  1.09 1.10 0.16 2.00  hatch)  Source  DF  SS  MS  f_  Total Treatment (A) v s . (B+C+D) (B) v s . (C+D) (C) v s . (D) Error  11 3 1 1 1 8  1146.9 604.6 2.2 420.5 181.5 542.3  201.5 2.2 420.5 181.5 67.8  2.97 0.03 6.20* 2.68  A n a l y s i s of  v a r i a n c e (sex X t r e a t m e n t ,  e f f e c t s on l o n g e v i t y )  Source  DF  SS  Total Treatment (A) v s . (B+C+D) (B) v s . (C+D) (C) v s . (D) Cages/treatment Sex Sex X t r e a t m e n t Sex X c a g e / t r e a t m e n t I n s e c t s / s e x and cage  95 (3) 1 1 1 8 1 3 8 72  1321.9 56.4 36.1 5.1 15.2 102.8 0.0 80.4 1014.6 33.9  MS  :  18.8 36.1 5.1 15.2 12.9 0.0 26.8 14.1 ' 4.2  F 1 .46 2.81 0.40 1.18 0.00 6.34*  -82-  Table XIII. Treatment 0 ppm d i f  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t c o d l i n g moths fed d i f l u b e n z u r o n s o l u t i o n s on c o t t o n w i c k s . Four males and f o u r females per cage. eggs total percent Cage Sex L o n g e v i t y (days) hatched eggs hatch X 1 2 3 1 2 3  100 ppm d i f  11 2 3  M F M F M F  9 , 14, 16, 16 8, 9 , 10, 10 7, 9, 12, 15 5, 8 , 9 , 13 8, 11, 12, 15 10, 12, 13, 14  493  567  86.9  427  530  80.6  343  447  76.7  M F M F M F  9, 8, 8, 7, 8, 9,  12, 10, 10, 9, 10, 13,  14, 11, 11, 9, 14, 14,  16 14 14 17 16 17  496  626  79.2  236 ,  315  74.9  641  749  85.6  M F M F M F  8, 7, 8, 9, 7, 9,  10, 9, 10, 9, 11, 10,  12, 16, 14, 12, 14, 12,  14 16 15 17 16 14  453  647  70.0  375  552  67.9  287  610  47.0  A n a l y s i s of variance  DF_  Total Treatment Linear regression Deviations Error  8 2 1 1 ,6  Source Total Treatment Linear regression Deviations Error  79.9  61 .6  (fecundity)  Source  A n a l y s i s of variance  81.4  SS 123952 11734 9129 2615 112217  MS .0 .9 .2 .2 .1  5867 .5 9129 .2 2615 .2 18702 .8  F_ 0 31 0 .49 0 02  (fecundity) DF 8 2 1 1 6  SS  MS  1176.2 737.8 738.7 0.2 438.4  368.9 738.7 0.2 73.1  F_ 5.05 10.12* 0.002  -83-  Table  x 111 ( c o n c l u d e d )  A n a l y s i s o f v a r i a n c e (sex X c o n c e n t r a t i o n , e f f e c t s on l o n g e v i t y ) Source  DF  Total Concentration Linear regression Deviations Cages/concentration Sex Sex X l i n e a r cone. Sex X d e v i a t i o n s cone. Sex X t o t a l cone. Sex X c a g e / t r e a t m e n t I n s e c t s / s e x and cage  71 (2) 1 1 6 1 1 1 (2) 6 54  Table Xiv. Treatment 0 ppm  1.0 ppm  10 ppm  100 ppm  1,000 ppm  SS  MS_  663.8 5.9 1.6 4.3 -36.9 9.4 14.1 0.9 13.4 557.0 41.3  f_  2.9 1.6 • 4.2 6.2 9.4 14.1 0.9 6.7 10.3 6.9  0.48 0.27 0.69 1.37 2.05 0.14 0.97  H a t c h a b i l i t y o f 0- t o 3 - d a y - o l d l e a f r o l l e r egg masses t o p i c a l l y treated with diflubenzuron solutions. One egg mass = one r e p l i c a t e . Rep.  eggs hatched  1 2 3 4 5  0 0 290 216 0  142 54 498 385 125  0.0 0.0 58.2 56.1 0.0  22.9  1 2 3 4 5  376 444 0 54 0  520 533 148 160 116  72.3 83.3 0.0 33.7 0.0  37.9  1 2 3 4 5  301 0 460 36 42  456 48 567 295 414  66.0 0.0 81.1 12.2 10.1  33.9  1 2 3 4 5  469 442 397 0 329  550 550 512 308 490  85.3 80.4 77.5 0.0 67.1  62.1  1 2 3 4 5  0 288 0 93 314  385 502 561 189 427  0.0 57.4 0.0 49.2 73.5  36.0  total  eggs  percent  hatch  X  -84-  T a b l e XIV  (concluded)  A n a l y s i s o f v a r i a n c e ( e x c l u d i n g 0 ppm) Source  DF  SS  MS  Total Treatment log Linear regression log Quadratic regression log Deviations Error  19 3 1 1 1 16  18958..8 2601..2 130..0 605..0 1866..2 16357..6  876..1 130..0 605..0 1866..2 1022..4  Individual  Table XV.  degree o f freedom c o n t r a s t :  f_ 0,.86 0,.13 0,.59 1..83  0 ppm v s . o t h e r t r e a t m e n t s F v a l u e = 1.50  -  S u r v i v a l o f f i r s t - i n s t a r l e a f r o l l e r l a r v a e p l a c e d on pear f o l i a g e which had been dipped i n d i f l u b e n z u r o n / T w e e n 20 s o l u t i o n s . Three l a r v a e per l e a f , one l e a f = one r e p l i c a t e .  Treatment 0 ppm d i f +  0 ppm Tw20 0 ppm d i f +  500 ppm Tw20 10 ppm. d i f 0- ppm Tw20 10 ppm d i f +  500 ppm Tw20 100 ppm d i f +  0 ppm Tw20 100 ppm d i f +  500 ppm Tw20  Rep.  Survival  a f t e r 1 week  Survival  a f t e r 2 weeks  1 2 3  1 2nd i n s t a r 2 2nd 1 3rd  1 3rd 2 3rd 1 4th  1 2 3  3 2nd 2 2nd 3 2nd  2 3rd, 1 4th 1 3rd, 1 4th 2 4th  1 2 3  3 2nd 2 2nd 2 2nd  2 3rd, 1 4th 2 3rd 1 4th  1 -2 3  3 2nd 2 2nd 2 2nd  2 3rd 1 4th 1 3 r d , 1 4th  3. 2nd . . . . 2 2nd 2 2nd- ... .  2 3rd 1 4th 1 3 r d , 1 4th  a l l dead 1 2nd a l l dead  a l l dead 2nd a i l dead  1 2 3 1 2 3  i  fl-  -85-  T a b l e XV  (concluded)  Analysis of variance  (on s u r v i v a l a f t e r 2 weeks)  Source  DF  SS.  Total Treatment D i f l u b e n z u r o n (A) Tween 20 (B) . (A) X (B) i n t e r a c t i o n Error  17 5 2 1 2 12  12.44 7.14 2.77 0.22 4.12 5.30  1  Table ; XVI. Treatment  :  MS 1.43 ••. 1.38 , 0.22 2.06 , 0.44  3.24* 3.14* 0.50 4.66*  M o u l t i n g success o f f i v e l e a f r o l l e r l a r v a l i n s t a r s p l a c e d on a p p l e f o l i a g e which had been sprayed w i t h d i f l u b e n z u r o n / T w e e n 20 solutions. One l a r v a per p l a n t , f i v e p l a n t s per t r e a t m e n t . :  1st i n s t a r  0 ppm d i f + 0 ppm Tw20  5  0 ppm d i f • +• 500 ppm Tw20  3  100 ppm d i f + 500 ppm Tw20  2  :  2nd i n s t a r  3rd i n s t a r  4th i n s t a r  6th  instar  3  3  5  4  4  4  3  5  1  2  4  3  -86-  Table XVII. Treatment  L o n g e v i t y , f e c u n d i t y and egg v i a b i l i t y o f a d u l t l e a f r o l l e r moths dipped i n d i f l u b e n z u r o n s o l u t i o n s . Four males and f o u r females per cage. eggs t o t a l percent Cage Sex L o n g e v i t y (days) hatched • eggs hatch X  0 ppm d i f  1 2 3 4  ppm d i f  1 2 3 4  8, 8 7, 9 7, 10 9, 9 8, 8 9 , 10 7, 8 6, 9  M F M F M F M F M F M F M F M F  2, 2, 1, 6, 1, 3, .. 3, 4, 2, 3, 2 , 3 , 2, 3, 5, 6,  Analysis of variance Source Total Treatment Error  6, 8, 6, 5, 5, 6, 3, 6,  Total Treatment Error Analysis of variance  1214  48.2  417  986  42.3  402  820  49.0  674  1143  59.0  411  726  56 .6  599  1088  55 .1  256  760  33 .7  522  827  63 .1  8 9 7 6 8 6 8 8  49.6  52.1  (fecundity)  7 1 6  DF 7 1 6  F  MS  SS  DF  A n a l y s i s of variance (percent Source  585  245428.0 72580.5 172847.5  72580. 5 28807. 9  2.52  hatch)  642.9 20.5 622.4  (sex X t r e a t m e n t ,  F  MS  SS  20.5 103.7  0.20  e f f e c t s on l o n g e v i t y )  Source  DF  SS  MS  Total Diflubenzuron Cages/cone. Sex Sex X t r e a t m e n t Sex X c a g e / t r e a t m e n t I n s e c t s / sex and cage  63 1 6 1 1 6 48  409.0 25.0 6.2 6.2 1.0 9.0 361 .5  25.0 1.0 6.2 1.0 1.5 7.5  IF 24.04* 4.17 0.67  -87-  Table < XVIII.  P e r c e n t o f h a r v e s t e d a p p l e s showing l e a f r o l l e r damage. s u b p l o t ( o f t h r e e t r e e s ) per t r e a t m e n t and b l o c k . '  Treatment  Tree  Block 1  Block 2  Block 3  Block 4  One  Block 5  X  0 ppm d i f + 0 ppm Tw20 (A)  1 2 3  16.0 19.0 24.7  15.7 15.4 16.1  12.0 8.4 6.6  12.1 26.6 15.2'  13.5 15.0 20.6  15.8  94 ppm d i f + 0 ppm Tw20 (B)  1 2 3  15.9 10.3 13.8  10.7 11.6 20.5  9.0 12.7 14.5  13.3 9.9 7.9  7.3 12.8 11.7  12.1  94 ppm d i f + 500 ppm Tw20 (C)  1 2 3  10.9 10.0 11.5  21.7 10.9 8.9  5.8 3.9 6.1  11.1 6.4 11.1  8.8 10.1 15.3  10.2  375 ppm d i f + 0 ppm Tw20 (D)  1 2 3  7.4 3.4 6.3  7.6 9.8 3.4  13.5 10.6 9.8  9.5 12.3 11.1  5.0 5.1 4.0  7.9  375 ppm azinphosmethyl (E)  1 2 3  1.2 2.7 1.1  0.4 0.7 2.7  2.6 1.9 2.2  1.6 1.0 0.6  3.7 4.0 2.2  1.9  A n a l y s i s of  variance  Source  Df_  SS  MS  F  Total Treatment (A) v s . (B+C+D+E) (B) v s . (C) (B) v s . (D) (D) v s . (E) Blocks B l o c k s X Treatment Error  74 4 1 1 1 1 4 16 50  2608.8 1606.1 715.5 24.1 132.7 271.2 60.1 418.4 524.2  401 .5 715 .5 24 .1 132 .7 271 .2 15 .0 26 .2  38.23* 68.14* 2.30 12.64* 25.83* 1 .43 2.50*  Table XIX.  P e r c e n t o f h a r v e s t e d a p p l e s showing c o d l i n g moth damage. One s u b p l o t ( o f t h r e e t r e e s ) o f each e u l t i v a r was p r e s e n t each t r e a t m e n t  Treatment  Tree  Spartan  Mcintosh  Delicious  Newtown  X  0.0 5.6 0.0  2.0  0.7 0.9 1 .0  0.8  47 ppm d i f + ' 0 ppm Tw20 (A)  1 2 3  0.3 0.0 0.5  4.0 0.6 1.6  47 ppm d i f + 500 ppm Tw20 (B)  1 2 3  0.7 0.7 0.5  1.1 1 .7 0.0  187 ppm d i f + 0 ppm Tw20 (C)  1 2 3  0.2 0.4 0.4  0.0 0.6 1.4  0.4 0.2 0.7  0.4  187 ppm azi.nphosmethyl (D)  1 2 3  0.8 0.2 0.2  2.3 0.0 1.1  0.7 0.8 0.5  0.7  Analysis of  0.5 0.2 1.2  •  variance  Source  DF  Total Cells Treatment (A) v s . (A) v s . (C) v s . Cultivar Treatment Within c e l l s  47 15 3 1 1 1 3 9 32=  (B) (C) (D) X Cultivar (error)  SS 83.95 36.28 17.15 8.17 14.88 0.57 5.93 13.20 47.67  MS  5.72 •8.'17;.  14.88 0i57. 1.47 1.49  F  3.84* 5.48* 9.99* 0.38 1.32 0.99  -89-  Table XX.  P e r c e n t o f h a r v e s t e d Golden D e l i c i o u s a p p l e s showing c o d l i n g moth damage. S u b p l o t s ( t h r e e t r e e s ) were arranged i n a balanced i n c o m p l e t e b l o c k d e s i g n w i t h f o u r t r e a t m e n t s randomized w i t h i n each b l o c k .  Treatment  Block 1  Block 2  Block 3  Block 4  Block 5  X  1 2 3  _  2.8 2.3 1.4  1.4 1.9 1.8  3.6 2.4 2.5  2.2 2.1 1.2  2.1  1 2 3  0.0 1.1 0.7  1 .3 0.6 1.1  1.3 0.5 0.7  0.4 1.2 1.1  -  0.8  1 2 3  0.9 0.0 0.8  1.1 1.2 0.5  _  -  0.8 1.1 1.0  0.6 1.4 0.6  0.8  1 2 3  0.0 0.5 0.0  _  -  0.6 1.0 0.2  0.0 1.1 0.4  0.7 0.8 0.8  0.5  1 2 3  0.0 0.0 0.0  0.7 0.7 0.8  0.5 0.6 1.2  -  0.6 0.0 0.3  0.4  Tree  0 ppm d i f +  0 ppm Tw20 (A) 47 ppm d i f +  0 ppm Tw20 (B) 47 ppm d i f +  500 ppm Tw20  (c5  187 ppm d i f +  0 ppm Tw20 (D) 187 ppm azinphosmethyl (E) A n a l y s i s of  _  .  variance  Source  DF  SS  Total Cells Treatments Treat (adjusted) Blocks Blocks (adjusted) (A) v s . (B+C+D+E) (B) v s . (C) (B) v s . (D) . (D) v s . (E) Exp. e r r o r Trees w i t h i n c e l l s  59 19 (4) 4 (4) 4 1 1 T 1 11 40  34.21 27.13 22.47 17.89 6.84 2.26 2.70 0.0004 0.43 0.008 2.40 7.08  MS  F_  4.47  20.52*  0.57 2.70 0.0004 0.43 0.008 0.22 0.18  3.20* 12.40* 0.002 1.96 0.04  

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