3
T h e Effect o f Estradiol Cypionate (ECP) on Ovarian Follicular Development a n d Ovulation i n Dairy Cattle Jacob Thundathil, P . Kastelic, a n d Reuben J . Mapletoft ABSTRACT Th e efficacy of estradiol cypi- onate (ECP) f o r synchroniznrg ovarian follicular development w a s deter- mined i n lactating Holstein-Friesian cattle. I n Experiment 1 , 1 3 cattle were given simultaneous intramus- cular (IM) injections o f 1 0 0 mg pro- gesterone a n d 0 (control), 0 . 5 o r 1 . 0 mg ECP o n D ay 3 , after a synchronized ovulation (Day 0 ) . Maximum diam- eter o f t h e dominant follicle o f Wave 1 w a s significantly larger i n control cattle than i n those given 0 . 5 o r 1 . 0 mg ECP (means: 15.7, 13.2, a n d 12.9 mm, respectively). Mean d a y o f emergence o f Wave 2 w a s significantly later i n controls than i n those given 1 . 0 mg ECP, with t h e 0. 5 mg group intermediate (Days 10.2, 8 . 8 a n d 9.5, respectively). I n Experiment 2 , 1 4 cattle were given a CIDR-B a n d I M injections o f 1 g ECP a n d 5 0 mg progesterone with- o u t regard t o stage o f cycle (treat- ment = D a y 0 ) . On D a y 8 , t h e CIDR-B w a s removed a nd 5 0 0 ,g clo- prostenol injected, I M . Mean days o f wave emergence (Day 3.4; range: - 2 t o 7 ) a n d ovulation (Day 12.1; range: 1 0 t o 1 4 ) indicated that ECP h a d limited efficacy f o r synchroniz- i n g follicular development a n d ovu- lation i n dairy cattle when given a t random stages o f t h e estrous cycle. RESUME L'efficacite d u cypionate d'estra- diol (CPE) pour l a synchronization d u developpement folliculaire ovarien chez l a vache Holstein- Friesian en lactation a e t e deter- mine. Dans l a premiere experience, 1 3 vaches o n t recu simultanement p a r voie intramusculaire (IM) 1 0 0 mg d e progesterone e t 0 (groupe temoin), 0 , 5 o u , 0 mg d e CPE a u jour 3 suivant u n e ovulation syn- chronisee (jour 0 ) . L e diametre maximal d u follicule dominant d e l a premiere vague folliculaire etait significativement plus gros chez l e s animaux temoins q u e chez ceux ayant recu 0 , 5 o u 1 , 0 mg d e CPE (moyennes d e 15,7, 13,2, et 12,9 mm , respectivement). L e jour moyen d'apparition d e l a deuxieme vague folliculaire etait significativement retarde pour l e groupe temoin com- parativement a u groupe ayant recu 1 ,0 mg d e CPE, alors q u e l e groupe ayant requ 0 , 5 mg d e CP E etait interme'diaire (jour 10,2, 8,8, e t 9,5, respectivement). Dans l a deuxieme experience, 1 4 vaches o n t recu un CIDR-B e t d e s injections I M d 1 mg d e CPE e t 5 0 mg d e prog- esterone sans egard a u stade o u elle s e trouvait dans leur cycle estral (jour d u traitement = jour 0 ) . Au jour 8 suivant l e traitement, l e CIDR-B f u t retire e t 5 0 0 , g d e clo- prostenol administre I . L e jour moyen d'apparition d e l a vague fol- liculaire (jour 3,4; etendue: - 2 a 7 ) e t d'ovulation (jour 12,1; etendue: 1 0 ' a 1 4 ) montre q u e l e CPE a u n e efficacite limitee pour synchroniser l e developpement folliculaire e t l'ovulation chez la vache laitiere lorsqu'il est administre a u hazard a u cours d u cycle cestral. (Traduit p a r docteur Serge Messier) Several estrus synchronization regi- mens include treatment with exoge- nous estrogens (e.g., estradiol valer- ate, estradiol benzoate o r estradiol-17B). Historically, estrogens were included i n synchronization regimens to induce luteal regression (1). However, estro- gens have recently been shown t o cause atresia o f antral follicles a n d effec- tively synchronize t h e emergence o f a ne w follicular wave (2). Th e only estrogen licensed f o r u s e i n cattle in Canada i s estradiol cypionate (ECP, T h e Upjohn Company, Orangeville, ON). To o u r knowledge, t h e u s e o f ECP i n a n estrus synchronization reg- imen h a s never been reported. Tw o experiments were conducted t o determine t h e efficacy o f ECP f o r synchronizing ovarian follicular development i n cattle. T h e objective o f Experiment 1 w a s t o determine the efficacy o f 2 different doses o f ECP (injected 3 d after ovulation) f o r inducing atresia o f t h e dominant folli- o f t h e first follicular wave (DF1) a n d f o r synchronizing emergence o f th e second follicular wave. T h e objec- tive o f Experiment 2 w a s t o determine t h e synchrony o f ovarian follicular wave emergence a n d ovulation fol- lowing ECP treatment a t random stages o f t h e estrous cycle ( i n cattle implanted with a CIDR-B). I n both experiments, cattle were utilized i n accordance with t h e guidelines o f t h e Canadian Council o n Animal Care (3 ) a n d with t h e approval o f t h e Leth- bridge Animal Care Committee. I n Experiment 1 , 1 3 lactating Holstein-Friesian cattle were used. T h e average number o f lactations w a s 1 . 8 (range: 1 t o 4 ) a n d t h e average interval from calving t o t h e start o f t h e experiment w a s 1 3 9 d (range: 4 0 t o 215). Transrectal ultrasonography w a s conducted with a B-mode diag- nostic ultrasound scanner with a Department o f Herd Medicine and Theriogenology, Western College o f Veterinary Medicine, University o f Saskatchewan, Saskatoon, Saskatchewan S7 N 5 B 4 (Thundathil, Mapletoft); Research Centre, Agriculture a n d Agri-Food Canada, Lethbridge, Alberta T1J 4B 1 (Kastelic). Correspondence t o : D r . J.P. Kastelic, Lethbridge Research Centre, Box 3000, Lethbridge, AB T I J 4B1; Telephone: (403) 317-2236; Fax: (403) 382-3156; e-mail: [email protected] Lethbridge Centre Contribution N o . 3879732 Received February 8 , 1998. C a n J V e t R e s 1997; 6 1 : 314-316 1 4

The Effect of Estradiol Cypionate on Ovarian Follicular Development in Dairy Cattle

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T h e E f f e c t o f E s t r a d i o l C y p i o n a t e ( E C P ) on O v a r i a n F o l l i c u l a rD e v e l o p m e n t a n d O v u l a t i o n i n D a i r y C a t t l e

J a c o b T h u n d a t h i l , J o h n P . K a s t e l i c , a n d R e u b e n J . M a p l e t o f t

ABSTRACT

Th e e f f i c a c y o f e s t r a d i o l c y p i -

o n a t e ( E C P ) f o r s y n c h r o n i z n r g o v a r i a n

f o l l i c u l a r d e v e l o p m e n t was d e t e r -

m i n e d i n l a c t a t i n g H o l s t e i n - F r i e s i a n

c a t t l e . I n E x p e r i m e n t 1 , 1 3 c a t t l ew e r e g i v e n s i m u l t a n e o u s i n t r a m u s -

c u l a r ( I M ) i n j e c t i o n s o f 1 0 0 mg p r o -

g e s t e r o n e a n d 0 ( c o n t r o l ) , 0 . 5 o r 1 . 0 mgECP o n Day 3 , a f t e r a s y n c h r o n i z e d

o v u l a t i o n ( D a y 0 ) . Maximum d i a m -e t e r o f t h e d o m i n a n t f o l l i c l e o f

Wave 1 w a s s i g n i f i c a n t l y l a r g e r i nc o n t r o l c a t t l e t h a n i n t h o s e g i v e n 0 . 5o r 1 . 0 mg ECP ( m e a n s : 1 5 . 7 , 1 3 . 2 ,a n d 1 2 . 9 mm, r e s p e c t i v e l y ) . M e and a y o f emergence o f Wave 2 w a ss i g n i f i c a n t l y l a t e r i n c o n t r o l s t h a ni n t h o s e g i v e n 1 . 0 mg E C P , w i t h t h e

0 . 5 mg g r o u p i n t e r m e d i a t e ( D a y s1 0 . 2 , 8 . 8 a n d 9 . 5 , r e s p e c t i v e l y ) . I n

E x p e r i m e n t 2 , 1 4 c a t t l e w e r e g i v e n a

CIDR-B a n d I M i n j e c t i o n s o f 1 mg

ECP a n d 5 0 mg p r o g e s t e r o n e w i t h -o u t r e g a r d t o s t a g e o f c y c l e ( t r e a t -m e n t = D a y 0 ) . On D a y 8 , t h e C I D R - Bw a s r emove d a nd 5 0 0 ,g c l o -p r o s t e n o l i n j e c t e d , I M . Mean d a y so f wave e m e r g e n c e ( D a y 3 . 4 ; r a n g e :

-2 t o 7 ) a n d o v u l a t i o n ( D a y 1 2 . 1 ;r a n g e : 1 0 t o 1 4 ) i n d i c a t e d t h a t ECPh a d l i m i t e d e f f i c a c y f o r s y n c h r o n i z -i n g f o l l i c u l a r d e v e l o p m e n t a n d o v u -

l a t i o n i n d a i r y c a t t l e when g i v e n a trandom s t a g e s o f t h e e s t r o u s c y c l e .

RESUME

L ' e f f i c a c i t e d u c y p i o n a t e d ' e s t r a -d i o l ( C P E ) p o u r l a s y n c h r o n i z a t i o ndu d e v e l o p p e m e n t f o l l i c u l a i r e

o v a r i e n c h e z l a v a c h e H o l s t e i n -F r i e s i a n e n l a c t a t i o n a e t e d e t e r -

m i n e . D a n s l a p r e m i e r e e x p e r i e n c e ,

1 3 v a c h e s o n t r e c u s i m u l t a n e m e n tp a r v o i e i n t r a m u s c u l a i r e ( I M )1 0 0 mg d e p r o g e s t e r o n e e t 0 ( g r o u p e

t e m o i n ) , 0 , 5 o u 1 , 0 mg d e CPE a uj o u r 3 s u i v a n t u n e o v u l a t i o n s y n -

c h r o n i s e e ( j o u r 0 ) . Le d i a m e t r em a x i m a l d u f o l l i c u l e d o m i n a n t d e l ap r e m i e r e v a g u e f o l l i c u l a i r e e t a i ts i g n i f i c a t i v e m e n t p l u s g r o s c h e z l e sanimaux t e m o i n s q u e c h e z c e u x

a y a n t r e c u 0 , 5 o u 1 , 0 mg d e CPE( m o y e n n e s d e 1 5 , 7 , 1 3 , 2 , e t 1 2 , 9 mm,r e s p e c t i v e m e n t ) . L e j o u r moyend ' a p p a r i t i o n d e l a d e u x i e m e v a g u ef o l l i c u l a i r e e t a i t s i g n i f i c a t i v e m e n tr e t a r d e p o u r l e g r o u p e t e m o i n c o m -

p a r a t i v e m e n t a u g r o u p e a y a n t r e c u1 , 0 mg d e C P E , a l o r s q u e l e g r o u p e

a y a n t r e q u 0 , 5 mg d e CPE e t a i ti n t e r m e ' d i a i r e ( j o u r 1 0 , 2 , 8 , 8 , e t 9 , 5 ,r e s p e c t i v e m e n t ) . D a n s l a d e u x i e m ee x p e r i e n c e , 1 4 v a c h e s o n t r e c u unCIDR-B e t d e s i n j e c t i o n s I M d e

1 mg d e CPE e t 5 0 mg d e p r o g -e s t e r o n e s a n s e g a r d a u s t a d e o u e l l es e t r o u v a i t d a n s l e u r c y c l e e s t r a l

( j o u r du t r a i t e m e n t = j o u r 0 ) . Auj o u r 8 s u i v a n t l e t r a i t e m e n t , l eCIDR-B f u t r e t i r e e t 5 0 0 , g d e c l o -p r o s t e n o l a d m i n i s t r e I M . Le j o u rmoyen d ' a p p a r i t i o n d e l a v a g u e f o l -l i c u l a i r e ( j o u r 3 , 4 ; e t e n d u e : - 2 a 7 )e t d ' o v u l a t i o n ( j o u r 1 2 , 1 ; e t e n d u e :1 0 ' a 1 4 ) m o n t r e q u e l e CPE a u n e

e f f i c a c i t e l i m i t e e p o u r s y n c h r o n i s e rl e d e v e l o p p e m e n t f o l l i c u l a i r e e t

l ' o v u l a t i o n c h e z l a v a c h e l a i t i e r el o r s q u ' i l e s t a d m i n i s t r e a u h a z a r da u c o u r s d u c y c l e c e s t r a l .

( T r a d u i t p a r d o c t e u r S e r g e M e s s i e r )

S e v e r a l e s t r u s s y n c h r o n i z a t i o n r e g i -mens i n c l u d e t r e a t m e n t w i t h e x o g e -n o u s e s t r o g e n s ( e . g . , e s t r a d i o l v a l e r -

a t e , e s t r a d i o l b e n z o a t e o r e s t r a d i o l - 1 7 B ) .H i s t o r i c a l l y , e s t r o g e n s w e r e i n c l u d e d

i n s y n c h r o n i z a t i o n r e g i m e n s t o i n d u c e

l u t e a l r e g r e s s i o n ( 1 ) . H o w e v e r , e s t r o -g e n s h a v e r e c e n t l y b e e n s h o w n t o c a u s e

a t r e s i a o f a n t r a l f o l l i c l e s a n d e f f e c -t i v e l y s y n c h r o n i z e t h e e m e r g e n c e o f a

ne w f o l l i c u l a r wave ( 2 ) . Th e o n l y

e s t r o g e n l i ce n s e d f o r u s e i n c a t t l e i nC a n a d a i s e s t r a d i o l c y p i o n a t e ( E C P ,T h e U p j o h n C o m p a n y , O r a n g e v i l l e ,

O N ) . To o u r k n o w l e d g e , t h e u s e o fECP i n a n e s t r u s s y n c h r o n i z a t i o n r e g -i m e n h a s n e v e r b e e n r e p o r t e d .Tw o e x p e r i m e n t s w e r e c o n d u c t e d

t o d e t e r m i n e t h e e f f i c a c y o f ECP f o rs y n c h r o n i z i n g o v a r i a n f o l l i c u l a r

d e v e l o p m e n t i n c a t t l e . T h e o b j e c t i v e

o f E x p e r i m e n t 1 w a s t o d e t e r m i n e t h ee f f i c a c y o f 2 d i f f e r e n t d o s e s o f ECP( i n j e c t e d 3 d a f t e r o v u l a t i o n ) f o r

i n d u c i n g a t r e s i a o f t h e d o m i n a n t f o l l i -c l e o f t h e f i r s t f o l l i c u l a r wave ( D F 1 )a n d f o r s y n c h r o n i z i n g e m e r g e n c e o f

t h e s e c o n d f o l l i c u l a r w a v e . T h e o b j e c -t i v e o f E x p e r i m e n t 2 w a s t o d e t e r m i n e

t h e s y n c h r o n y o f o v a r i a n f o l l i c u l a rwave e m e r g e n c e a n d o v u l a t i o n f o l -l o w i n g ECP t r e a t m e n t a t randoms t a g e s o f t h e e s t r o u s c y c l e ( i n c a t t l ei m p l a n t e d w i t h a C I D R - B ) . I n b o t h

e x p e r i m e n t s , c a t t l e w e r e u t i l i z e d i na c c o r d a n c e w i t h t h e g u i d e l i n e s o f t h eC a n a d i a n C o u n c i l o n A n i m a l C a r e ( 3 )a n d w i t h t h e a p p r o v a l o f t h e L e t h -

b r i d g e A n i m a l C a r e C o m m i t t e e .

I n E x p e r i m e n t 1 , 1 3 l a c t a t i n g

H o l s t e i n - F r i e s i a n c a t t l e w e r e u s e d .T h e a v e r a g e n u m b e r o f l a c t a t i o n s w a s

1 . 8 ( r a n g e : 1 t o 4 ) a n d t h e a v e r a g e

i n t e r v a l f r o m c a l v i n g t o t h e s t a r t o f

t h e e x p e r i m e n t w as 1 3 9 d ( r a n g e : 4 0t o 2 1 5 ) . T r a n s r e c t a l u l t r a s o n o g r a p h ywas c o n d u c t e d w i t h a B-mode d i a g -n o s t i c u l t r a s o u n d s c a n n e r w i t h a

D e p a r t m e n t o f H e r d M e d i c i n e a n d T h e r i o g e n o l o g y , W e s t e r n C o l l e g e o f V e t e r i n a r y M e d i c i n e , U n i v e r s i t y o f S a s k a t c h e w a n , S a s k a t o o n , S a s k a t c h e w a n

S7 N 5 B 4 ( T h u n d a t h i l , M a p l e t o f t ) ; R e s e a r c h C e n t r e , A g r i c u l t u r e a n d A g r i - F o o d C a n a d a , L e t h b r i d g e , A l b e r t a T 1 J 4B 1 ( K a s t e l i c ) .

C o r r e s p o n d e n c e t o : D r . J . P . K a s t e l i c , L e t h b r i d g e R e s e a r c h C e n t r e , Box 3 0 0 0 , L e t h b r i d g e , AB T I J 4 B 1 ; T e l e p h o n e : ( 4 0 3 ) 3 1 7 - 2 2 3 6 ; F a x : ( 4 0 3 ) 3 8 2 - 3 1 5 6 ;

e - m a i l : k a s t e l i c @ e m . a g r . c aL e t h b r i d g e R e s e a r c h C e n t r e C o n t r i b u t i o n N o . 3 8 7 9 7 3 2

R e c e i v e d F e b r u a r y 8 , 1 9 9 8 .

C a n J V e t R e s 1 9 9 7 ; 6 1 : 3 1 4 - 3 1 61 4

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7 . 5 MHz t r a n s d u c e r ( D y n a m i cI m a g i n g , M o d e l MCV, L i v i n g s t o n ,

S c o t l a n d , U K ) . J u s t p r i o r t o t h e s t a r to f t h e e x p e r i m e n t , a l l c a t t l e h a d a n

u l t r a s o n i c a l l y d e t e c t a b l e c o r p u sl u t e u m a n d n o n e h a d u l t r a s o n i c a l l yd e t e c t a b l e p a t h o l o g y o f t h e u t e r u s o ro v a r i e s . A l l c a t t l e w e r e g i v e n a n IM

i n j e c t i o n o f 5 0 0 , u g c l o p r o s t e n o l( E s t r u m a t e , M a l l i n c k r o d t V e t e r i n a r y

I n c . , A j a x , ON) t o i n d u c e l u t e a lr e g r e s s i o n . T h e r e a f t e r , u l t r a s o u n d

e x a m i n a t i o n s w e r e p e r f o r m e d o n c e

d a i l y t o c o n f i r m o v u l a t i o n ( D a y 0 ) .T h e c a t t l e w e r e r a n d o m l y a l l o c a t e d t o3 g r o u p s a n d g i v e n t h e f o l l o w i n g b yI M i n j e c t i o n o n D a y 3 : 1 0 0 mg p r o g -

e s t e r o n e ( S i g m a C h e m i c a l C o . L t d . ,S t . L o u i s , MO) i n s e s a m e o i l ( c o n t r o lg r o u p , n = 5 ) ; 1 0 0 mg p r o g e s t e r o n ea n d 0 . 5 mg ECP ( n = 4 ) ; o r 1 0 0 mg

p r o g e s t e r o n e a n d 1 . 0 mg ECP ( n = 4 ) .I n j e c t i o n s o f p r o g e s t e r o n e a n d ECPw e r e g i v e n a t s e p a r a t e l o c a t i o n s .

P r o g e s t e r o n e w a s i n j e c t e d t o p r e v e n t

a n e s t r o g e n - i n d u c e d l e u t i n i z i n g h o r -

mone s u r g e . D a i l y u l t r a s o u n d e x a m i -

n a t i o n s w e r e c o n d u c t e d f r o m D a y 0 t oDay 1 2 , a n d t h e s i z e a n d l o c a t i o n o f

f o l l i c l e s : 4 mm w e r e r e c o r d e d . T h ee m e r g e n c e o f Wave 1 a n d Wave 2

w e r e d e f i n e d a s t h e s y n c h r o n o u sg r o w t h o f s e v e r a l f o l l i c l e s , 4 t o 5 mmi n d i a m e t e r ( 4 ) . A n a l y s i s o f v a r i a n c e

( 5 ) w a s u s e d t o d e t e r m i n e i f t h e r e w a sa n e f f e c t o f t r e a t m e n t g r o u p o n t h ed a y o f e m e r g e n c e o f e a c h w a v e a n do n t h e maximum d i a m e t er o f D F I . AL e a s t S i g n i f i c a n t D i f f e r e n c e ( L S D )t e s t w a s u s e d t o d e t e c t d i f f e r e n c e s . AG e n e r a l L i n e a r M o d e l s (GLM)r e p e a t e d - m e a s u r e s a n a l y s i s o f v a r i -a n c e ( 5 ) w a s u s e d t o d e t e r m i n e t h e

e f f e c t o f t r e a t m e n t , t i m e , a n d t h e

t r e a t m e n t - b y - t i m e i n t e r a c t i o n f o r t h ed i a m e t e r o f DF1 o n D a y s 1 t o 1 2 .

I n E x p e r i m e n t 1 , t r e a t m e n t w i t h

ECP e f f e c t i v e l y s u p p r e s s e d t h e

g r o w t h o f D F 1 . T h e e f f e c t o f t r e a t -m e n t g r o u p o n wave e m e r g e n c e w a s

s i g n i f i c a n t f o r Wave 2 ( P < 0 . 0 3 ) b u t

n o t f o r Wave 1 ( T a b l e I ) . Wave 2

e m e r g e d o n D a y s 9 t o 1 1 ( c o n t r o l s ) ,D a y s 9 o r 1 0 ( 0 . 5 mg E C P ) , o r D a y s 8o r 9 ( 1 . 0 mg E C P ) . Mean d a y o f e m e r -

g e n c e o f Wave 2 wa s s i g n i f i c a n t l ye a r l i e r i n c a t t l e g i v e n 1 . 0 mg ECPt h a n i n c o n t r o l c a t t l e ( D a y 8 . 8 v s

Day 1 0 . 2 ) . H o w e v e r , e m e r g e n c e o f

Wave 2 i n c a t t l e t r e a t e d w i t h 0 . 5 mg( D a y 9 . 5 ) w a s n o t s i g n i f i c a n t l y d i f f e r -e n t f r o m t h e o t h e r t w o g r o u p s . T h e r e

TABLE I . Mean ( ± s t a n d a r d d e v i a t i o n ) d a y o f e m e r g e n c e o f Waves 1 a n d 2 a n d maximumd i a m e t e r o f t h e d o m i n a n t f o l l i c l e o f Wave 1 ( D F 1 ) i n c a t t l e g i v e n p r o g e s t e r o n e ( 1 0 0 m g ) a n de s t r a d i o l c y p i o n a t e ( E C P ) o n Day 3 ( o v u l a t i o n = Day 0 ) i n E x p e r i m e n t 1

ECP ( m g )

E n d p o i n t 0 0 . 5 1 . 0

N o . o f c a t t l e 5 4 4

Wave I e m e r g e n c e ( D a y ) 0 . 2 + 0 . 8 - 0 . 8 + 0 . 5 0 . 6 ± 0 . 8Maximum d i a m e t e r D F 1 ( m m ) a 1 5 . 7 + 1 . 8 b 1 3 . 2 + 0 . 9 c 1 2 . 9 + 0 . 8 c

Wave 2 e m e r g e n c e ( D a y ) d 1 0 . 2 + 0 . 8 b 9 . 5 + 0 . 6 b c 8 . 8 + 0 . 5 ca E f f e c t o f t r e a t m e n t ( P < 0 . 0 2 )k W i t h i n a r o w , m e a n s w i t h o u t a common s u p e r s c r i p t a r e d i f f e r e n t ( P < 0 . 0 5 )d E f f e c t o f t r e a t m e n t ( P < 0 . 0 3 )

w a s a n e f f e c t o f t r e a t m e n t o n m a x i -mum d i a m e t e r o f D FI ( P < 0 . 0 2 ) ,

w i t h a s i g n i f i c a n t l y g r e a t e r d i a m e t e ri n t h e c o n t r o l g r o u p t h a n i n t h e o t h e r2 g r o u p s . F o r t h e d i a m e t e r o f D F I o n

D a y s 1 t o 1 2 ( F i g u r e 1 ) , t h e r e w a sa n e f f e c t o f t i m e ( P < 0 . 0 0 0 1 ) a n da t r e a t m e n t - b y - t i m e i n t e r a c t i o n

( P < 0 . 0 0 4 ) b u t t h e e f f e c t o f t r e a t m e n tw a s n o t s i g n i f i c a n t ( P > 0 . 4 ) .Th e r e s u l t s o f E x p e r i m e n t 1

a p p e a r e d p r o m i s i n g a n d w e r e t h e

i m p e t u s f o r c o n d u c t i n g E x p e r i m e n t 2 .I n E x p e r i m e n t 2 , 1 5 l a c t a t i n g

H o l s t e i n - F r i e s i a n c a t t l e w e r e u s e d .

T h e a v e r a g e n u m b e r o f l a c t a t i o n s w a s2 . 1 ( r a n g e : 1 t o 4 ) a n d t h e a v e r a g ei n t e r v a l f r o m c a l v i n g t o t h e s t a r t o f

t h e e x p e r i m e n t w a s 9 8 . 8 d ( r a n g e : 5 4

t o 2 1 3 ) . A l l e x a m i n a t i o n s w e r e c o n -

d u c t e d w i t h t r a n s r e c t a l u l t r a s o n o g r a -

p h y , a s d e s c r i b e d i n E x p e r i m e n t 1 .None o f t h e c a t t l e h a d u l t r a s o n i c a l l y

2 0 7

E

c o

0

1 5 -

1 0

5 .

0

d e t e c t a b l e p a t h o l o g y o f t h e u t e r u s o r

o v a r i e s . On t h e f i r s t d a y o f t h e e x p e r i -m e n t ( d e s i g n a t e d a s Day 0 ) , a l l o f t h ec a t t l e h a d a n u l t r as o n i c a l l y v i s i b l ec o r p u s l u t e u m ( b u t t h e s p e c i f i c d a y o f

t h e c y c l e w a s u n k n o w n ) . A C o n t r o l l e d

I n t e r n a l D r u g R e l e a s e d e v i c e ( C I D R - B ,

V e t r e p h a r m C a n a d a I n c . , L o n d o n ,

ON) w a s i n s e r t e d i n t r a v a g i n a l l y a n dt h e c a t t l e w e r e g i v e n 1 mg ECP a n d5 0 mg p r o g e s t e r o n e ( s a m e s o u r c e s a si n E x p e r i m e n t 1 ) b y I M i n j e c t i o n i ns e p a r a t e l o c a t i o n s . The CIDR-Bd e v i c e c o n t a i n s 1 . 9 g p r o g e s t e r o n e

t h a t i s r e l e a s e d a n d a b s o r b e d ,i n c r e a s i n g s e r u m p r o g e s t e r o n e c o n -

c e n t r a t i o n s . On Day 8 , t h e CIDR-Bw a s r e m o v e d a n d 5 0 0 , g c l o p r o s t e n o l

was i n j e c t e d I M . T r a n s r e c t a l u l t r a -s o u n d e x a m i n a t i o n s w e r e c o n d u c t e d( a s d e s c r i b e d i n E x p e r i m e n t 1 ) o n c e

d a i l y f r o m Day 0 t o t h e d a y o f o v u l a -t i o n f o l l o w i n g c l o p r o s t e n o l t r e a t m e n t .

_~~~~~a.aO'e._, , #

mu

',d of

F ~ ~ ~ ~ ~ C o n t r o l---ECP 0 . 5----ECP 1 . 0

0 5 1 0 1 5

D a yF i g u r e 1 . M e an d i a m e t e r o f t h e d o m i n a n t f o l l i c l e o f Wave 1 on D a y s 1 t o 1 2 ( o v u l a t i o n =

D a y 0 ) i n 1 3 c a t t l e i n E x p e r i m e n t 1 . On D a y 3 , a l l c a t t l e were g i v e n 1 0 0 mg p r o g e s t e r o n e a n d

t h e f o l l o w i n g d o s e s o f ECP ( e s t r a d i o l c y p i o n a t e ) : 0 mg ( C o n t r o l ) , 0 . 5 mg ( E C P 0 . 5 ) , o r 1 . 0 mg(ECP 1 . 0 ) . T h e r e w a s a n e f f e c t o f t i m e ( P < 0 . 0 0 0 1 ) and a t r e a t m e n t - b y - t i m e i n t e r a c t i o n

( P < 0 . 0 0 5 ) , b u t t h e e f f e c t o f t r e a t m e n t wa s n o t s i g n i f i c a n t .

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I n t h i s e x p e r i m e n t , o v u l a t i o n was

n o t d e t e c t e d b y Day 1 4 i n 1 cow a n d

s h e w a s r e m o v e d f r o m t h e s t u d y . I na n o t h e r c o w , a n o v a r i a n f o l l i c l e t h a tw a s 8 mm i n d i a m e t e r o n Day 0 c o n -

t i n u e d t o g r o w , b e c a m e t h e d o m i n a n t

f o l l i c l e , a n d r e a c h e d a maximumd i a m e t e r o f 2 3 mm p r i o r t o o v u l a t i o n

o n D a y 1 0 . I n t h i s c o w , Wave 1 w a s

e s t i m a t e d t o h a v e s t a r t e d o n D a y - 2 .

I n E x p e r i m e n t 2 , t h e r e w a s c o n s i d -

e r a b l e v a r i a b i l i t y among c a t t l e i n f o l -l i c u l a r d y n a m i c s , w i t h r e l a t i v e l y p o o r

s y n c h r o n y o f wave e m e r g e n c e a n d

o v u l a t i o n ( T a b l e I I ) . Wave e m e r g e n c e

r a n g e d f r o m D a y - 2 t o Day 7 a n d o v u -

l a t i o n o c c u r r e d o n D a y s 1 0 , 1 1 , 1 2 ,1 3 , o r 1 4 i n 1 , 5 , 3 , 4 , a n d 1 a n i m a l s ,r e s p e c t i v e l y . T h e r e f o r e , t h e r e wasi n s u f f i c i e n t s y n c h r o n y o f w a v e e m e r -

g e n c e a n d o v u l a t i o n t o w a r r a n t t h eu s e o f ECP i n a n e s t r u s s y n c h r o n i z a -

t i o n p r o g r a m . H o w e v e r , i t ma y b eu s e f u l i f a d d i t i o n a l t r e a t m e n t i s g i v e n

t o s y n c h r o n i s e o v u l a t i o n .A l t h o u g h e s t r a d i o l - 1 7 B ( E - 1 7 8 ) i s

t h e m o s t p o w e r f u l n a t u r a l e s t r o g e n i nm a m m a l s , i t h a s a s h o r t h a l f - l i f e ( 6 ) .I n o n e s t u d y ( 7 ) , a n IM i n j e c t i o n o f

5 mg E - 1 7 8 r e s u l t e d i n p e a k p l a s m a

e s t r o g e n c o n c e n t r a t i o n s b y 6 h a f t e rt r e a t m e n t ( t i m e o f f i r s t p o s t - t r e a t m e n t

b l o o d s a m p l e ) , w i t h a d e c l i n e t o b a s e -

l i n e c o n c e n t r a t i o n s b y 4 2 h . E s t e r s o fE - 1 7 B h a v e a much l o n g e r h a l f - l i f e .F o r e x a m p l e , w h e n e s t r a d i o l v a l e r a t ewas i n j e c t e d I M i n c a t t l e , p l a s m ae s t r a d i o l c o n c en t ra t io n s w e r e e l e v a t e d

w i t h i n 1 2 t o 2 4 h a f t e r t r e a t m e n t a n d

r e m a i n e d a b o v e b a s e l i n e c o n c e n t r a -

t i o n s f o r a p p r o x i m a t e l y 1 2 0 h ( 8 ) .T r e a t m e n t w i t h a v e r y l a r g e d o s e

( 1 0 mg) o f ECP r e s u l t e d i n serumE - 1 7 B c o n c e n t r a t i o n s p e a k i n gb e t w e e n 1 3 a n d 3 1 h a f t e r t r e a t m e n t ,a n d r e m a i n i n g a b o v e maximum p h y s i -

o l o g i c a l c o n c e n t r a t i o n s ( d e f i n e d a s

maximum c o n c e n t r a t i o n s p r i o r t o

e s t r u s ) f o r a n a v e r a g e o f 1 7 0 h a f t e rt r e a t m e n t ( 6 ) . T h e d o s e s u s e d i n t h ep r e s e n t e x p e r i m e n t s ( 1 . 0 a n d 0 . 5 m g )w e r e 1 0 o r 2 0 t i m e s l o w e r t h a n t h a tu s e d i n t h e p r e v i o u s s t u d y ( 6 ) .A l t h o u g h e s t e r i f i c a t i o n p r o l o n g s t h e

h a l f - l i f e o f E - 1 7 B , i t a p p e a r s t o r e d u c e

i t s e f f i c a c y f o r s y n c h r o n i z a t i o n o f f o l -

TABLE I I . M e a n , s t a n d a r d d e v i a t i o n ( S D ) a n d r a n g e f o r f o l l i c l e d i a m e t e r , wave e m e r g e n c ea n d o v u l a t i o n f o r 1 4 c a t t l e g i v e n 1 . 0 mg e s t r a d i o l c y p i o n a t e ( E C P ) , 5 0 mg p r o g e s t e r o n e a n d a

CIDR-B ( d e s i g n a t e d a s D a y 0 ) . On D a y 8 , t h e CIDR-B w a s r e m o v e d a n d c l o p r o s t e n o l ( 5 0 0 , u g )w a s g i v e n ( E x p e r i m e n t 2 )

M e a s u r e m e n t Mean SD Minimum Maximum

Wave e m e r g e n c e ( D a y ) 3 . 4 2 . 1 - 2 7

D o m i n a n t f o l l i c l e d i a m e t e r , D a y 8 ( m m ) 1 2 . 6 3 . 9 6 2 3D i a m e t e r p r e - o v u l a t i o n ( m m ) 1 6 . 7 2 . 2 1 4 2 3

O v u la t i o n ( D a y ) 1 2 . 1 1 . 0 1 0 1 4Wave e m e r g e n c e t o o v u l a t i o n ( d ) 8 . 5 1 . 6 6 1 2

l i c u l a r d e v e l o p m e n t a n d o v u l a t i o n .

T r e a t m e n t w i t h E - 1 7 B c o n s i s t e n t l y

s u p p r e s s e d o v a r i a n f o l l i c l e s , w i t h

s y n c h r o n o u s emergence o f a ne ww a v e , o n a v e r a g e , 4 . 3 d a f t e r t r e a t -ment ( 2 , 7 , 8 ) . I n c o n t r a s t , e s t r a d i o l

v a l e r a t e s u p p r e s s e d f o l l i c l e s l e s s c o n -

s i s t e n t l y a n d e m e r g e n c e o f a new f o l -l i c u l a r w a v e s e e m e d l e s s s y n c h r o n o u s

( 9 ) , s i m i l a r t o t h e r e s p o n s e i n E x p e r i -

m e n t 2 .I n c o n c l u s i o n , w h e n l a c t a t i n g d a i r y

c a t t l e w e r e g i v e n 0 . 5 o r 1 . 0 mg ECP( p l u s 1 0 0 mg p r o g e s t e r o n e ) 3 d a f t e ro v u l a t i o n , g r o w t h o f DF1 was s u p -

p r e s s e d a n d e m e r g e n c e o f Wave 2 w a s

s y n c h r o n o u s . H o w e v e r , w h e n 1 . 0 mgECP ( p l u s 5 0 mg p r o g e s t e r o n e a n d a

C I D R - B ) w e r e g i v e n a t r a n d o m s t a g e s

o f t h e e s t r o u s c y c l e , wave e m e r g e n c e

was a s y n c h r o n o u s ( r a n g e 2 d b e f o r e

t r e a t m e n t t o 7 d a f t e r t r e a t m e n t ) . F u r -

t h e r m o r e , f o l l o w i n g CIDR-B r e m o v a la n d t r e a t m e n t w i t h c l o p r o s t e n o l ( 8 d

a f t e r ECP t r e a t m e n t ) , o v u l a t i o n was

p o o r l y s y n c h r o n i z e d ( r a n g e : 2 t o 6 d

a f t e r CIDR-B r e m o v a l ) . T h e r e f o r e ,t h e p r o l o n g e d h a l f - l i f e o f ECPa p p e a r s t o m a k e i t f a r l e s s e f f i c a c i o u st h a n E - 1 7 1 7 f o r s y n c h r o n i z i n g f o l l i c u -l a r wave e m e r g e n c e a n d o v u l a t i o n i nd a i r y c a t t l e .

ACKNOWLEDGMENTS

The s u p p o r t o f A g r i c u l t u r e a n dA g r i - F o o d C a n a d a , t h e C a n a d a / A l b e r t a

L i v e s t o c k R e s e a r c h T r u s t , I n c . , a n d

T h e U n i v e r s i t y o f S a s k a t c h e w a n , a r e

g r a t e f u l l y a c k n o w l e d g e d . W e t h a n k

M a l l i n c k r o d t V e t e r i n a r y I n c . , f o r p r o -

v i d i n g E s t r u m a t e , V e t r e p h a r mC a n a d a , I n c . , f o r p r o v i d i n g CIDR-Bd e v i c e s , B y r n e Cook a n d M a r c e l oM a r t i n e z f o r a s s i s t a n c e w i t h d a t a

a n a l y s i s , F r a n c e s C o t e f o r w o r d p r o -

c e s s i n g , a n d S h e i l a T o r g u n r u d f o r

g r a p h i c s .

REFERENCES

1 . ODDE KG. A r e v i e w o f s y n c h r o n i z a t i o no f e s t r u s i n p o s t p a r t u m c a t t l e . J Anim S c i1 9 9 0 ; 6 8 : 8 1 7 - 8 3 0 .

2 . BO G , ADAMS G P , PIERSON RA AN DMAPLETOFT R J . Exogenous c o n t r o l

o f f o l l i c u l a r wave e m e r g e n c e i n c a t t l e .T h e r i o g e n o l o g y 1 9 9 5 ; 4 3 : 3 1 - 4 0 .

3 . CANADIAN COUNCI L ON ANIMAL

CARE. G u i d e t o t h e C a r e a n d Us e o f

E x p e r i m e n t a l A n i m a l s , 2 n d e d . V o l 1 .O t t a w a : B r a d d a P r i n t i n g S e r v i c e s , 1 9 9 3 .

4 . GINTHER O J , KASTELIC J P , KNOPFL . C o m p o s i t i o n a n d c h a r a c t e r i s t i c s o f f o l -l i c u l a r w a v e s d u r i n g t h e b o v i n e e s t r o u s

c y c l e . Anim R e p r o d S c i 1 9 8 9 ; 2 0 :

1 8 7 - 2 0 0 .

5 . SAS/STAT USER'S GUIDE. V e r s i o n 6 .F o u r t h E d i t i o n , V o l u m e 1 . SAS I n s t i t u t e ,C a r y , N C , 1 9 9 0 ; 8 9 0

p p .6 . VYNCKIER L , DEBACKERE M, DE

KRUIF A , CORYN M. P l a s m a e s t r a d i o l -1 7 B c o n c e n t r a t i o n s i n t h e c ow d u r i n gi n d u c e d e s t r u s a n d a f t e r i n j e c t i o n o f

e s t r a d i o l - 1 7 1 3 b e n z o a t e a n d e s t r a d i o l - 1 7 1 3c y p i o n a t e- p r e l i m i n a r y s t u d y . J V e t

P h a r m a c o l T h e r a p 1 9 9 0 ; 1 3 : 3 6 - 4 2 .7 . BO GA, ADAMS GP, PIERSON RA,

TRIBULO HE, CACCIA M, MAPLE.TOFT R J . F o l l i c u l a r w a v e d y n a m i c s a f t e re s t r a d i o l - 1 7 1 7 t r e a t m e n t o f h e i f e r s w i t h o r

w i t h o u t a p r o g e s t o g e n i m p l a n t . T h e r i -

o g e n o l o g y 1 9 9 4 ; 4 1 : 1 5 5 5 - 1 5 6 9 .

8 . BO G , ADAMS G P , CACCIA M, MAR-TINEZ M, PIERSON RA, MAPLE-

TOFT R J . O v a r i a n f o l l i c u l a r w a v e e m e r -g e n c e a f t e r t r e a t m e n t w i t h p r o g e s t a g e n a n d

e s t r a d i o l i n c a t t l e . A n i m a l R e p r o d u c t i o nS c i e n c e 1 9 9 5 ; 3 9 : 1 9 3 - 2 0 4 .

9 . BO GA, ADAMS G P , NASSER L F ,PIERSON R A , MAP L ETO F T R J . E f f e c to f e s t r a d i o l v a l e r a t e o n o v a r i a n f o l l i c l e s ,e m e r g e n c e o f f o l l i c u l a r w a v e s a n d c i r c u -l a t i n g g o n a d o t r o p i n s i n h e i f e r s . T h e r i -

o g e n o l o g y 1 9 9 3 ; 4 0 : 2 2 5 - 2 3 9 .

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