The Effect of o,p'-DDT 1 on Japanese Quail by A. S. CooKs Monks Wood Experimental Station, Abbots Ripton, Huntingdon, England
T e c h n i c a l DDT i n c r e a s e s u t e r i n e w e t w e i g h t in r a t s (1). The m o s t a c t i v e c o n s t i t u e n t is not p , p ' - D D T 2 but t h e o , p ' i s o m e r , w h i c h , it w a s s u g g e s t e d , c o u l d c o m p e t e w i t h l a b e l l e d o e s t r a d i o l for b i n d i n g s i t e s on t h e u t e r u s . It h a s now b e e n r e p o r t e d (2) t h a t a d m i n i s t r a t i o n of o , p ' - D D T c a n c a u s e o e s t r o g e n i c - t y p e r e s p o n s e s in t h e w e i g h t and g l y c o g e n c o n t e n t of c h i c k e n and q u a i l o v i d u c t s a n d rat u t e r i . If o,p'-DDT is a potent oestrogen, large scale treatment of the environment could produce a serious hazard since the commercial D D T mixture contains 11-29% of the o,p'-isomer (3). Oestrogens increase serum calcium and lipid levels in birds (4) and the effect of o,p'-DDT on Japanese quail (Coturnix coturnix japonica) has been studied, paying particular attention to any changes in these parameters. Experimental methods E x p e r i m e n t 1~ Two g r o u p s of 6 m a l e , a n d two of 6 f e m a l e , J a p a n e s e q u a i l (age 33 d a y s , w e i g h t 6 0 - 8 0 g) w e r e r a n d o m l y s e l e c t e d . Each bird in o n e group of e a c h s e x w a s t r e a t e d d a i l y for 4 d a y s w i t h o , p ' - D D T 3 (5 rag in 0 . 2 ml o l i v e o i l , by i n t r a m u s c u l a r i n j e c t i o n ) . Birds in t h e c o n t r o l group w e r e i n j e c t e d
1 o , p' - D D T : 1,1, 1 - t r i c h l o r o - 2 - ( o - c h l o r o p h e n y ) - 2 (p-chlorophenyl) ethane 2p, p. - D D T : 1, 1, l - t r i c h l o r o - 2 , 2 - b i s ( p - c h l o r o p h e n y l ) e t h a n e 3Aldrich C h e m i c a l C o . , M i l w a u k e e , W i s . 152 Bulletin of Environmental Contamination & Toxicology, Vol. 5, No. 2, 1970, published by Springer-Verlag New York, Inc.
w i t h o l i v e o i l o n l y . Blood s a m p l e s w e r e t a k e n from t h e j u g u l a r v e i n o f a n a e s t h e t i s e d f e m a l e b i r d s 72 h o u r s a f t e r t h e f i n a l i n j e c t i o n , a n d f r o m t h e m a l e s 24 h o u r s l a t e r . Birds w e r e s a c r i f i c e d a n d e i t h e r o v i d u c t a n d o v a r y w e i g h t or t e s t e s w e i g h t r e c o r d e d . S a m p l e s of b r e a s t m u s c l e w e r e a n a l y s e d for organochlorine residues. E x p e r i m e n t 2. Six a d u l t m a l e q u a i l (age 73 d a y s , w e i g h t 94-108 g) w e r e e a c h o r a l l y d o s e d b y m e a n s of a c a t h e t e r , w i t h l0 mg o , p ' - D D T in 0 . 2 ml o l i v e o i l , 3 d o s e s p e r w e e k b e i n g a d m i n i s t e r e d for 3 w e e k s . A f u r t h e r g r o u p of 6 q u a i l w a s s i m i l a r l y t r e a t e d w i t h o l i v e o i l o n l y . Blood s a m p l e s w e r e w i t h d r a w n from t h e b r a c h i a l v e i n p r i o r to d o s i n g a n d a g a i n a t t h e e n d of t h e 3 w e e k p e r i o d . After t h e s e c o n d b l o o d s a m p l e w a s taken, birds were sacrificed and liver and testes were weighed. L i v e r s a m p l e s w e r e a n a l y s e d for o r g a n o c h l o r i n e r e s i d u e s . A n a l y s i s . All s e r u m s a m p l e s w e r e a n a l y s e d for t o t a l c a l c i u m (5) a n d t h o s e from t h e f e m a l e s w e r e a n a l y s e d for t o t a l l i p i d (6). T i s s u e s a m p l e s for o r g a n o c h l o r i n e a n a l y s i s w e r e g r o u n d w i t h sand and anhydrous sodium sulphate and extracted with hot, r e d i s t i l l e d n - h e x a n e , f o l l o w e d by r e d i s t i l t e d a c e t o n e . Extracts were passed through an alumina column and residues determined by gas-liquid chromatography using an electron capture detector.
Results Results for Experiments 1 and 2 are given in Tables 1 and 2 respectively. In both experiments the m e a n testes weight of the treated birds w a s less than that of the corresponding controls ,but because of variation within the groups, neither difference is statistically significant. No significant differences occurred between respective treated and control groups for any of the organs studied. In Experiment 2 serum calcium levels for birds in the o , p ' - D D T group were virtually the same before and after treatment, while in Experiment I, m e a n levels for the treated birds were slightly higher than for the control birds, but the differences were not significant. The percentage gain in weight of females treated with o , p ' - D D T w a s lower (P
T e c h n i c a l DDT i n c r e a s e s u t e r i n e w e t w e i g h t in r a t s (1). The m o s t a c t i v e c o n s t i t u e n t is not p , p ' - D D T 2 but t h e o , p ' i s o m e r , w h i c h , it w a s s u g g e s t e d , c o u l d c o m p e t e w i t h l a b e l l e d o e s t r a d i o l for b i n d i n g s i t e s on t h e u t e r u s . It h a s now b e e n r e p o r t e d (2) t h a t a d m i n i s t r a t i o n of o , p ' - D D T c a n c a u s e o e s t r o g e n i c - t y p e r e s p o n s e s in t h e w e i g h t and g l y c o g e n c o n t e n t of c h i c k e n and q u a i l o v i d u c t s a n d rat u t e r i . If o,p'-DDT is a potent oestrogen, large scale treatment of the environment could produce a serious hazard since the commercial D D T mixture contains 11-29% of the o,p'-isomer (3). Oestrogens increase serum calcium and lipid levels in birds (4) and the effect of o,p'-DDT on Japanese quail (Coturnix coturnix japonica) has been studied, paying particular attention to any changes in these parameters. Experimental methods E x p e r i m e n t 1~ Two g r o u p s of 6 m a l e , a n d two of 6 f e m a l e , J a p a n e s e q u a i l (age 33 d a y s , w e i g h t 6 0 - 8 0 g) w e r e r a n d o m l y s e l e c t e d . Each bird in o n e group of e a c h s e x w a s t r e a t e d d a i l y for 4 d a y s w i t h o , p ' - D D T 3 (5 rag in 0 . 2 ml o l i v e o i l , by i n t r a m u s c u l a r i n j e c t i o n ) . Birds in t h e c o n t r o l group w e r e i n j e c t e d
1 o , p' - D D T : 1,1, 1 - t r i c h l o r o - 2 - ( o - c h l o r o p h e n y ) - 2 (p-chlorophenyl) ethane 2p, p. - D D T : 1, 1, l - t r i c h l o r o - 2 , 2 - b i s ( p - c h l o r o p h e n y l ) e t h a n e 3Aldrich C h e m i c a l C o . , M i l w a u k e e , W i s . 152 Bulletin of Environmental Contamination & Toxicology, Vol. 5, No. 2, 1970, published by Springer-Verlag New York, Inc.
w i t h o l i v e o i l o n l y . Blood s a m p l e s w e r e t a k e n from t h e j u g u l a r v e i n o f a n a e s t h e t i s e d f e m a l e b i r d s 72 h o u r s a f t e r t h e f i n a l i n j e c t i o n , a n d f r o m t h e m a l e s 24 h o u r s l a t e r . Birds w e r e s a c r i f i c e d a n d e i t h e r o v i d u c t a n d o v a r y w e i g h t or t e s t e s w e i g h t r e c o r d e d . S a m p l e s of b r e a s t m u s c l e w e r e a n a l y s e d for organochlorine residues. E x p e r i m e n t 2. Six a d u l t m a l e q u a i l (age 73 d a y s , w e i g h t 94-108 g) w e r e e a c h o r a l l y d o s e d b y m e a n s of a c a t h e t e r , w i t h l0 mg o , p ' - D D T in 0 . 2 ml o l i v e o i l , 3 d o s e s p e r w e e k b e i n g a d m i n i s t e r e d for 3 w e e k s . A f u r t h e r g r o u p of 6 q u a i l w a s s i m i l a r l y t r e a t e d w i t h o l i v e o i l o n l y . Blood s a m p l e s w e r e w i t h d r a w n from t h e b r a c h i a l v e i n p r i o r to d o s i n g a n d a g a i n a t t h e e n d of t h e 3 w e e k p e r i o d . After t h e s e c o n d b l o o d s a m p l e w a s taken, birds were sacrificed and liver and testes were weighed. L i v e r s a m p l e s w e r e a n a l y s e d for o r g a n o c h l o r i n e r e s i d u e s . A n a l y s i s . All s e r u m s a m p l e s w e r e a n a l y s e d for t o t a l c a l c i u m (5) a n d t h o s e from t h e f e m a l e s w e r e a n a l y s e d for t o t a l l i p i d (6). T i s s u e s a m p l e s for o r g a n o c h l o r i n e a n a l y s i s w e r e g r o u n d w i t h sand and anhydrous sodium sulphate and extracted with hot, r e d i s t i l l e d n - h e x a n e , f o l l o w e d by r e d i s t i l t e d a c e t o n e . Extracts were passed through an alumina column and residues determined by gas-liquid chromatography using an electron capture detector.
Results Results for Experiments 1 and 2 are given in Tables 1 and 2 respectively. In both experiments the m e a n testes weight of the treated birds w a s less than that of the corresponding controls ,but because of variation within the groups, neither difference is statistically significant. No significant differences occurred between respective treated and control groups for any of the organs studied. In Experiment 2 serum calcium levels for birds in the o , p ' - D D T group were virtually the same before and after treatment, while in Experiment I, m e a n levels for the treated birds were slightly higher than for the control birds, but the differences were not significant. The percentage gain in weight of females treated with o , p ' - D D T w a s lower (P
