Wat. Re5. V01. 30, N0. 7, pp.

1711-1723, 1996
~ Per9am0n P11: 50043-1354(96)00054-1
C0pyr19ht • 1996 E15ev1er5c1enceLtd
Pr1nted 1n 6reat 8r1ta1n. A11r19ht5 re5erved
0043-1354/96 $15.00 + 0.00

ANALY515 0 F 7 0 X 1 C 1 7 Y 7 E 5 7 5 0 N DAPHN1A 5URV1VAL

AND REPR0DUC710N

5. A. L. M. K 0 0 1 J M A N * and J. J. M. 8 E D A U X
Vr1je Un1ver51te1t, de 80e1e1aan 1087, 1081 HV Am5terdam, 7he Nether1and5

(F1r5t rece1ved Apr11 1995; accepted 1n rev15edf0rm Fe6ruary 1996)

A65tract--We pre5ent a 5tat15t1ca1 ana1y515 0f 610a55ay5 f0r Daphn1a 5urv1va1 and repr0duct10n, 5uch a5
the r0ut1ne t0x1c1ty te5t that 15de5cr16ed 1n the 0 E C D 9u1de11ne 202. 7he ana1y51515 6a5ed 0n the Dynam1c
Ener9y 8ud9et the0ry and a 0ne-c0mpartment k1net1c5 f0r the t0x1c c0mp0und. 1t 15fu11ypr0ce55 0r1ented.
We c0mpare a f0rmu1at10n 1n term5 0f effect5 0n 5urv1va1 dur1n9 009ene515 t0 var10u5 d1rect and 1nd1rect
effect5 0n the ener9et1c5 0f repr0duct10n. A11 f0rma1at10n5 character12e the effect5 6y a n0-effect
c0ncentrat10n, a t01erance c0ncentrat10n and the e11m1nat10n rate. We c0nc1ude that a11 0pt10n5 1ead t0
51m11ar n0-effect 1eve15.We c0mpare the ana1y515 t0 the 5tandard N0EC/EC50 ana1y515 and c0nc1ude that
0ur ana1y515 15 60th 51mp1er and m0re effect1ve. C0pyr19ht • 1996 E15ev1er 5c1ence Ltd

Key w0rd5--NEC, 610a55ay5, repr0duct10n5, t01erance c0ncentrat10n

1N7R0DUC710N f0r th15 part1cu1ar te5t. H0wever, many feature5 app1y

t0 0ther te5t5 a5 we11. We f1r5t 5ummar15e the re1evant
70x1c1ty te5t5 are 0f 1ntere5t f0r 5c1ent1f1c purp05e5 a5 deta115 0f the te5t 9u1de11ne, then we w0rk 0ut a new
we11 a5 f0r 1e9151at10n and r15k a55e55ment purp05e5. ana1y515 0f the5e te5t5 and c0mpare th15 w1th the
7he n0-065erved-effect c0ncentrat10n ( N 0 E C ) 5t111 5tandard ana1y515.
p1ay5 an 1mp0rtant r01e 1n the 1atter purp05e5, 6ut 1t5
u5e 15 under 1ncrea5ed 5crut1ny 6ecau5e 0f a550c1ated
5tat15t1ca1 pr061em5. Ear11er attempt5 t0 501ve the5e R0U71NE 70X1C17Y 7E575
pr061em5 6y the 1nc0rp0rat10n 0f the n0-effect
7he r0ut1ne t0x1c1ty te5t 0n Daphn1a ma9na
c0ncentrat10n (NEC) a5 a m0de1 parameter 5uffer5
repr0duct10n acc0rd1n9 t0 9u1de11ne 202 0f the
fr0m d1ff1cu1t1e5, that w111 6e d15cu55ed 1ater.
0 E C D (1994) re4u1re5 that y0un9 fema1e5 ( < 1 d) are
5ma11-effect c0ncentrat10n5 (EC5, EC10) recent1y
1nd1v1dua11y exp05ed t0 a ran9e 0f c0ncentrat10n5 0f
9a1ned 1ntere5t, 6ut the5e character15t1c5 are a150 far
te5t c0mp0und dur1n9 a per10d 0f 21 d. 5urv1va1 and
fr0m 1dea1, a5 w111 6e exp1a1ned 1ater. 1n th15 art1c1e,
num6er 0f 0ff5pr1n9 are 065erved 0n a da11y 6a515
we pr0p05e a 501ut10n that 5eem5 free fr0m 5tat15t1ca1
when f00d (9reen a19ae 5uch a5 Ch10re11a 0r
and pract1ca1 pr061em5, 6ut 1t re4u1re5 a rad1ca1
5cenede5mu5) 15 5upp11ed. 7 h e med1a are renewed
chan9e 1n the way that we 100k at t0x1c effect5: a fu11y
three t1me5 a week. 7he temperature and c0ncen-
pr0ce55-6a5ed appr0ach.
trat10n 0f c0mp0und 1n the med1a are a5 c0n5tant a5
Repr0duct10n can 6e affected d1rect1y 6y t0x1c
p055161e (1n the ran9e 0f 18-22°C). At 1ea5t 10
chem1ca15, 0r 1nd1rect1y v1a effect5 0n feed1n9, 9r0wth
rep11cate5 are re4u1red f0r each ch05en te5t c0ncen-
0r ma1ntenance. 7h15 15 6ecau5e the5e pr0ce55e5 are
trat10n, 0ne 0f them 6e1n9 the c0ntr01.7he c0eff1c1ent
1nt1mate1y 11nked t0 each 0ther. 7he Dynam1c Ener9y
f0r the var1at10n 0f the num6er 0f 11v1n9 0ff5pr1n9 1n
8ud9et ( D E 8 ) the0ry pr0v1de5 a mechan15t1c 6a515
the c0ntr01 5h0u1d 6e 1e55 than 0.3.
that ha5 6een te5ted a9a1n5t many exper1menta1 data
7he fema1e daphn1d5 typ1ca11y 9r0w fr0m 0.8 mm
(K001jman, 1993). F19ure 1 91ve5 a 5ummary 0f the
t0 4 mm 6y day 21 dur1n9 exp05ure, wh1ch mean5 an
ener9y f10w5 1n an 0r9an15m that the D E 8 the0ry
1ncrea5e 6y a fact0r 5 1n 1en9th and a fact0r 125 1n
5pec1f1e5 4uant1tat1ve1y.
we19ht.
1n th15 art1c1e we pre5ent and app1y a 5tat15t1ca1
ana1y515 0f r0ut1ne t0x1c1ty te5t5 0n Daphn1a
repr0duct10n 6a5ed 0n 1n519ht5 fr0m the D E 8 the0ry. M0DEL5
51nce th15 te5t 15 the 6e5t 5tandard12ed am0n9 a11
A pract1ca1 pr061em f0r the ana1y515 0f repr0duc-
a4uat1c t0x1c1ty te5t5 and app11ed 0n a r0ut1ne 6a515
t10n data re5u1t1n9 fr0m a r0ut1ne t0x1c1ty te5t 15 that
a11 0ver the w0r1d, 1t 15 u5efu1 t0 w0rk 0ut an ana1y515
var1a61e5 5uch a5 feed1n9, 9r0wth and re5p1rat10n
rate5 are u5ua11y n0t mea5ured. 7h15 make5 1t hard t0
* Auth0r t0 wh0m c0rre5p0ndence 5h0u1d 6e addre55ed. 5e1ect the pathway f0r the effect 0n repr0duct10n that

1711
1712 5. A. L. M. K001jman and J. J. M. 8edaux

app11e5 t0 the c 0 m p 0 u n d te5ted. W e w111 5ee, h0wever,

t h a t 5evera1 p a r a m e t e r 5 t h a t character12e the t0x1c1ty a551m11at10n
0 f the c 0 m p 0 u n d d 0 n 0 t d e p e n d 5en51t1ve1y 0n the 7
5e1ect10n 0 f the c 0 r r e c t p a t h w a y . W e f1r5t d15cu55 1 re5erve5 ]
t0x1c0k1net1c5, t h e n repr0duct10n and f1na11y effect5,
6ecau5e we 11nk effect5 t0 t155ue-c0ncentrat10n5, n 0 t
env1r0nment-c0ncentrat10n5. 7 h e e4uat10n5 d1rect1y
~ ..repr0duct10n
9r0~1nt. ~.~
f0110w f r 0 m the D E 8 t h e 0 r y f0r c 0 n 5 t a n t f00d
den51ty.
F19. 1. 7he p0wer5 a5 5pec1f1ed 4uant1tat1ve1y 6y the DE8
6R0W7H m0de1 f0r an ec0therm w1th 60dy 512e and re5erve den51ty
a5 5tate var1a61e5.70x1c c0mp0und5 that affect repr0duct10n
A1th0u9h 6 0 d y 512e 15 n 0 t u5ua11y m e a 5 u r e d 1n can d0 50 d1rect1y, 0r 1nd1rect1y v1a a551m11at10n, 9r0wth and
ma1ntenance. 7he r0unded 60xe5 1nd1cate 50urce5 0r 51nk5.
r0ut1ne t0x1c1ty te5t5, 9 r 0 w t h ha5 t0 6e c0n51dered
6ecau5e t0x1c0k1net1c5 d e p e n d 5 0 n 6 0 d y 512e a n d
effect5 0 n 5urv1va1 a n d repr0duct10n d e p e n d 5 0 n the 6 0 d y 512e a n d ;) 15 the v0n 8erta1anffy 9 r 0 w t h rate.
a m 0 u n t 0 f the c 0 m p 0 u n d 1n the 60dy. N 0 t 6e1n9 7h15 rate p a r a m e t e r d e p e n d 5 0n tw0 ener9y
mea5ured, 6 0 d y 1en9th ha5 the r01e 0 f a h1dden p a r a m e t e r 5 a5 ~ = r h 9 / 3 ( f + 9), w h e r e the ma1nten-
var1a61e. 1t pr0ve5 t0 6e c0nven1ent t0 w 0 r k w1th ance rate c0eff1c1ent rh 5tand5 f0r the rat10 0 f the
5ca1ed 1en9th, r a t h e r t h a n actua11en9th, 1 =-- ( V / V m ) ~3, v01ume-5pec1f1c m a 1 n t e n a n c e c05t5 (per un1t 0 f t1me)
w h e r e V~ 315 the m a x 1 m u m v01umetr1c 1en9th an adu1t a n d the v01ume-5pec1f1c c05t5 f0r 9 r 0 w t h , 1.e. the
c a n reach. a m 0 u n t 0 f ener9y re4u1red t0 5ynthe512e a un1t
1f f00d den51ty 15 k e p t c0n5tant, the D E 8 t h e 0 r y v01ume 0 f 6 1 0 m a 5 5 . 7 h e 1nve5tment rat10 9 15 the rat10
5tate5 that 5ca1ed 1en9th 6ehave5 a5 0 f the v01ume-5pec1f1c c05t5 f0r 9 r 0 w t h a n d the
fract10n 0 f the re5erve den51ty that 15 a110cated t0
d
d-t 1 = ~ ( f - 1) (1) 9 r 0 w t h p1u5 ma1ntenance. 7 h e m a x 1 m u m v01umetr1c
1en9th d e p e n d 5 0 n ener9y p a r a m e t e r 5 a5 V),;3 = 1~/rh9,
w h e r e the ener9y c 0 n d u c t a n c e t~ repre5ent5 the rat10
w h e r e f 15 the 1n9e5t10n rate a5 a fract10n 0 f the
0 f the 5urface area-5pec1f1c a551m11at10n rate a n d the
m a x 1 m u m 1n9e5t10n rate f0r a n 1nd1v1dua1 0 f t h a t
v01ume-5pec1f1c c05t5 f0r 9 r 0 w t h . W e need the ener9y
1nterpretat10n5 0 f the v 0 n 8erta1anffy 9 r 0 w t h rate
7a61e 1. L15t 0f 5ym6015. 7he 5ym6015 1n the d1men510n-c01umn a n d the m a x 1 m u m v01umetr1c 1en9th 1n the d15cu5510n
5tand f0r: t, t1me: m, ma55; 1, 1en9th (0f env1r0nment); L, 1en9th (0f
0r9an15m); #, num6er 0 f 1nd1rect effect5 0 n repr0duct10n.
5ym601 D1men510n 1nterpretat10n 1f f00d den51ty 15 k e p t c0n5tant a n d 1f the
c m1-3 c0ncentrat10n 1n env1r0nment c 0 m p 0 u n d d0e5 n 0 t affect feed1n9, 9 r 0 w t h 0r
c4 m1 3 rat10 0f c0nc. 1n t155ue and P,a ma1ntenance, 5ca1ed 1en9th a5 a funct10n 0 f t1me
[Q] mL 3 c0ncentrat10n 1n t155ue a m 0 u n t 5 t0 the v0n 8erta1anffy 9 r 0 w t h curve
Pva P L -3 610c0ncentrat10n c0eff1c1ent
N0EC m1-3 n0-065erved effect 1(t) = f - - (f- 16) exp{ - ~,~t} (2)
c0ncentrat10n
c0 m1 3 n0-effect c0nc. f0r 5urv1va1, w h e r e /6 15 the 5ca1ed 1en9th at 61rth, 1.e. 1 ( 0 ) = 16.
repr0d.
CL50,CL50
~ m1-~ 50% effect c0nc. f0r 5urv., N0t1ce t h a t f0r a 6 u n d a n t f00d, we have that f = 1
u1t1mate cL50 a n d t h a t 1t 15 n 0 t 1mp0rtant that f00d den51ty 15
ce50 m1-3 50% effect c0ncentrat10n f0r repr0d. c0n5tant, a5 10n9 a5 f00d 15 a 6 u n d a n t . F 0 r D. m a 9 n a
f1, f1R 9rad1ent parameter f0r 5urv., repr0d.
c. m1-3 t01erancec0nc. f0r * ~ H, at 20°C a n d a 6 u n d a n t f00d we typ1ca11y f1nd 16 = 0.18
R,M, 60rA a n d "2 = 0.1 d -~. F19ure 3 91ve5 the typ1ca1 9 r 0 w t h
/,:* m • 13t-' k1111n9rate
/~° t ~ e11m1nat10nrate curve f0r D. m a 9 n a dur1n9 the 21 day5 0 f exp05ure.
4 -- t01erancefunct10n f0r 5urv1va1
5 5tre55 funct10n f0r repr0duct10n REPR0DUC710N
t t exp05uret1me
/~0./1~ t -~ ha2ard rate 1n c0ntr01, c0mp0und
1nduced E n e r 9 y a110Cat10n t0 repr0dUCt10n 15 a C0nt1nu0U5
f -- 1n9e5t10na5 fract10n 0f pr0Ce55, wh11e repr0dUCt10n 1t5e1f 15 e v e n t dr1ven. 7h15
1t5 max. 91ven 1 mean5 that ener9y a110Cated t0 repr0dUCt10n f1r5t
1 -- 60dy 1en9th a5 fract10n
0f 1t5 max. aCCUmU1ate5 1n a 6Uffer a n d that the C0ntent 0 f th15
16, 1p -- 1 at 61rth, pu6erty 6Uffer 15 1n5tantane0U51y C0nVerted 1nt0 (a C1UtCh 0f)
,• t-• v0n 8erta1anffy 9r0wth rate
rn r ~ ma1ntenance rate c0eff1c1ent e995. Daphn1a m0U1t5 at a fre4uenCy that d e p e n d 5 0n
~ Lt • ener9y c0nductance t e m p e r a t u r e . W h e n a C1UtCh 0 f hatCh11n95 15 de11vered,
9 ener9y 1nve5tment rat10 the fema1e m0U1t5, C0nVert5 the 6Uffer 0 f ener9y
-- 5urv1va1pr06a6111ty
/~,/~m # t ~ repr0d, rate. max. repr0d, rate a110Cated t0 repr0dUCt10n 1nt0 e995 a n d dep051t5
N # num6er 0f 0ff5pr1n9 fre5h1y 1a1d e995 1nt0 the 6r00dp0UCh. 50 the
,t~ t ~ p0pu1at10n9r0wth rate 1nterm0U1t per10d e4Ua15 the 1nCU6at10n t1me. 7he5e
 Daphn1a repr0duct10n t0x1c1ty te5t5 1713

deta115 are 5pec1f1c t0 Daphn1a. We f1r5t d15cu55 t0x1c UP7AKE/EL1M1NA710N K1NE71C5

effect5 0n repr0duct10n, treated a5 a c0nt1nu0u5
pr0ce55, 6ecau5e th15 app11e5 t0 a w1de var1ety 0f
an1ma15 and then d15cu55 deta115 that re1ate t0 the
c1utche5 0f Daphn1a.
Repr0duct10n depend5 0n 60dy 512e and ener9y
re5erve5, 50 1nd1rect1y 0n f00d ava11a6111ty. 7h15
mean5 that 1n a c0ntr01 51tuat10n, where an 0r9an15m
5uch a5 D. m a 9 n a 15 f0110wed 1nd1v1dua11y fr0m an a9e
1e55 than 1 d up t0 21 d at a c0n5tant 0r h19h f00d
den51ty, the repr0duct1ve eff0rt can 6e de5cr16ed 6y a
5at1at1n9 funct10n 0f a9e. Acc0rd1n9 t0 the D E 8
the0ry, the repr0duct10n rate at c0n5tant f00d den51ty
a5 a funct10n 0f 5ca1ed 1en9th 15
(3)
/
f0r 1p < 1 < f , where 1p 15 the 5ca1ed 1en9th at pu6erty,
wh1Ch am0unt5 t0 1p = 0.4 f0r D. ma9na, 7he va1ue
f0r the 1nve5tment rat10 f0r D. m a 9 n a 15 1e55 Certa1n,
6ut 9 "•" 1 15 rea115t1c. 1t m19ht 6e 50mewhat 5ma11er,
6ut numer1Ca1 5tud1e5 1nd1Cate that th15 hard1y affect5
t0x1C1ty parameter5. 5U65t1tut10n 0f the va1ue5
f0r 1p and 9 1nt0 e4uat10n (3) 91ve5
R(1) = 0.53R~,(12 + 13 -- 0.128). 7he parameter J~m
can 6e 1nterpreted a5 the max1mum repr0duct10n
rate, 1.e. the repr0duct10n rate 0f an 1nd1v1dua1 0f
max1mum 512e at a6undant f00d. 1t 15 pr0p0rt10na1 t0
ma1ntenance c05t5 -2, a551m11at10n rate 3 and 1ndepen-
dent 0f the 9r0wth c05t5. F19ure 2 111u5trate5 that
e4uat10n (3), t09ether w1th e4uat10n (2), f1t data we11.
208
168
120 >"
0
e0 0
A5 15 re1evant f0r the 0 E C D t0X1C1ty te5t W1th D.
m a 9 n a ( 0 E C D , 1994), exp05Ure 15 a55Umed t0 5tart at
61rth at a C0n5tant enV1r0nment-C0nCentrat10n C 0f
t0X1C C0mp0Und. We Ch005e t = 0 a5 the 5tart 0f the
exp05Ure per10d and the t1me at Wh1Ch 1 = 1p, 5ay 1p,
a5 the 5tart 0f the repr0dUCt1Ve eff0rt, 50 1(tp) = 1p.
5Upp05e that a650rpt10n 0f the C0mp0Und t0 the
f00d part1C1e5 15 1n5tantane0U5 and that the
C0nCentrat10n 0f f00d part1C1e5 15 C0n5tant. Uptake
Can 0CCUr d1rect1y fr0m the Water and 1nd1reCt1y V1a
f00d, 6Ut 60th Uptake rate5 are pr0p0rt10na1 t0 the
5UrfaCe area 0f the an1ma1, Wh1Ch 15 pr0p0rt10na1 W1th
V::3 f0r an 150m0rph 5UCh a5 Daphn1a. 7 h e d1rect
e11m1nat10n 15 a55Umed t0 6e pr0p0rt10na1 t0 the
5UrfaCe area a9a1n and t0 the C0nCentrat10n 1n the
a4Ue0U5 fract10n 0f the t155Ue [Q]. 7he part1t10n1n9 0f
the C0mp0Und 0Ver the d1fferent 60dy fract10n5,
1nC1Ud1n9 the 11p1d fract10n, 15 a55Umed t0 6e
1n5tantane0U5 a9a1n. 7he Uptake/e11m1nat10n k1net1C5
redUCe5 t0
d~ [Q] = cPvd11af/1 - [Q] 1~f/1 + -d-11n 13 (4)
where c 15 the c0ncentrat10n 1n the env1r0nment
(d15501ved p1u5 a650r6ed t0 f00d part1c1e5), Pvd 15 the
610c0ncentrat10n c0eff1c1ent and /~a the e11m1nat10n
rate. 7he term
ddt 1n P = 31-1 d 1
1n e4uat10n (4) acc0unt5 f0r the d11ut10n 6y 9r0wth.
7he t155ue-c0ncentrat10n 15 n0t u5ua11y mea5ured
1n r0ut1ne t0x1c1ty te5t5, 50 that 1t p1ay5 the r01e 0f
a h1dden var1a61e, ju5t 11ke 60dy 1en9th. 1t pr0ve5
t0 6e c0nven1ent t0 1ntr0duce the 5ca11n9 c4 = [Q]/Pvd,
wh1ch ha5 the d1men510n5 0f an env1r0nment-
c0ncentrat10n, 6ut 15 ju5t pr0p0rt10na1 t0 the
t155ue-c0ncentrat10n. 7he k1net1c5 0f the 5ca1ed
t155ue-c0ncentrat10n reduce5 t0

c4 = c1~,f/1 - c4 ~ 1n 13 (5)
U • • • •
40

• t1me, d
0 A1th0u9h th15 51mp1e f1r5t 0rder d1fferent1a1 e4uat10n
8 4 8 12 16 20 w1th var1a61e c0eff1c1ent5 can 6e 501ved, th15 hard1y
he1p5 6ecau5e the 501ut10n 5t111ha5 1nte9ra15 that mu5t
F19. 2. 7he cumu1at1ve num6er 0f y0un9 per fema1e D.
ma9na, 5tart1n9 fr0m 1nd1v1dua15 0f a9e ~<1 d w1th a 5ca1ed 6e 06ta1ned numer1ca11y. At a6undant f00d, where
1en9th 0f 0.18.7he data repre5ent a c0ntr01 fr0m a te5t that f = 1, the 6ehav10ur 0n1y depend5 0n the 5ca1ed
ha5 6een executed under 0 E C D 9u1de11ne 202.7he curve 15 e11m1nat10n rate /~, re1at1ve t0 the v0n 8erta1anffy
the 1ea5t 54uare5 f1t, a55um1n9 that the repr0duct10n rate 15 9r0wth rate ~;. F19ure 3 111u5trate5 h0w the 5hape 0f
11near 1n the 54uared and cu6ed 1en9th, wh11e 1en9th 9r0w5
w1th a v0n 8erta1anffy 9r0wth rate 0f 0.1 d ~. 7he 5ca1ed the uptake/e11m1nat10n curve depend5 0n the e11m1n-
1en9th at pu6erty 15 0.4. 7he e5t1mated max1mum at10n rate. 1t a150 5h0w5 that the pr0ce55 0f d11ut10n
repr0duct10n rate 15 28.9 d t . 6y 9r0wth 15 1mp0rtant f0r the k1net1c5.
1714 5. A. L. M. K001jman and J. J. M. 8edaux
c4/c
1 •t01erance•
0.8
0°6
0.4
0.2
{~ ~1 1 1 1 1 1 1 1 1
0 0.4 0.8 1 .2 1 .6
F19. 3.7he 5ca1ed t155ue-c0ncentrat10n f0r an 0r9an15m that
9r0w5 at c0n5tant f00d den51ty w1th v0n 8erta1anffy 9r0wth
rate "~. 7he uptake and e11m1nat10n rate fr0m the water 15
pr0p0rt10na1 t0 the 5urface area 0f the 0r9an15m. Uptake
and e11m1nat10n can 6e d1rect1y fr0m the water a5 we11a5 v1a
f00d. 7he hatch11n9 0r191nate5 fr0m a cu1ture 1n th15
examp1e. 7he f1ne1y 5t1pp1ed curve repre5ent5 the 5ca1ed
1en9th, the c0ar5e1y 5t1pp1ed curve 15the 5ca1ed repr0duct10n
rate, the 5011d curve5 the t155ue-c0ncentrat10n5 f0r d1fferent
ch01ce5 0f the e11m1nat10n rate /~a. 7he 5ca11n9 15 5uch that
a11 curve5 have a5ympt0te 1.7he parameter va1ue5 16 = 0.18,
1p = 0.5, 9 = 1 are typ1ca1 f0r D. ma9na at 20°C and t~; = 2
c0rre5p0nd5 w1th 21 d.
EFFEC75 0N 5URV1VAL
7he ha2ard rate depend5 11near1y 0n the t155ue-c0n-
centrat10n, thu5 a150 0n the 5ca1ed t155ue-c0ncen-
trat10n. 7he 1dea 6eh1nd th15 11near re5p0n5e funct10n
15 that each m01ecu1e wh1ch exceed5 the phy5101091ca1
capac1ty t0 hand1e the c0mp0und ha5 an effect that
15 1ndependent fr0m that 0f 0ther m01ecu1e5. We can
re1ax th15 5tr1ct 1nterpretat10n 6y 5tat1n9 that the
actua1 way 1n wh1ch the ha2ard rate 15 affected 15
c0mp1ex and n0n-11near, 6ut that we appr0x1mate the
de5cr1pt10n w1th tw0 term5 0f the 7ay10r expan510n
ar0und the n0-effect c0ncentrat10n. 7h15 appr0x1-
mat10n 15 0n1y va11d f0r 5ma11 effect5, 0f c0ur5e, 6ut
5ma11 effect5 0n the ha2ard rate (and 0n ener9y
parameter5 that w1116e d15cu55ed 1ater) end up hav1n9
c0n51dera61e effect5 0n 5urv1va1 (and repr0duct10n).
7he ha2ard rate and the 5urv1va1 pr06a6111ty are
91ven 6y
/~¢ =/~,(c4 - c0)+
~ ( c , t) = 4 ( c , t ) ~ ( 0 , t)
where c0 15 the n0-effect c0ncentrat10n (f0r 5urv1va1),
/~, the k1111n9 rate and Y(0, t ) = exp{-/~0t} 15 the
c0ntr01 5urv1va1 pr06a6111ty, w1th/~0 6e1n9 the ha2ard
k°/+ rate 1n the c0ntr01, wh1ch 15 c0n51dered t0 6e
c0n5tant. 7he funct10n 4(c, t) w111 6e ca11ed the
funct10n (f0r 5urv1va1). N0t1ce that
10
4(0, t ) = 1 15 the max1mum va1ue f0r the t01erance
funct10n. 7he 1ndex + mean5: (x)+ ~ max{0, x}.
57A71571C5
1
61ven 065ervat10n t1me5 {tt, t2 . . . . . tk} and te5t
c0ncentrat10n5 {ct, c2. . . . . cr}, the num6er 0f dead
0.5 (0r 1mm061112ed) 1nd1v1dua15 1n the t1me-1nterva1
( t ~ , td f0110w5 a mu1t1n0m1a1 d15tr16ut10n w1th
pr06a6111ty parameter 4(cj, t, ~) - 4(% t~). 51nce the
parameter5/~0,/~,, /~ and c0 0ccur 1n the pr06a6111ty
~.1
parameter5 0f a11 065ervat10n t1me5 and te5t-c0ncen-
trat10n5, a11 065ervat10n5 5h0u1d 6e c0n51dered
1 51mu1tane0u51y t0 extract the5e f0ur parameter5 u51n9
2 the max1mum 11ke11h00d meth0d (5ee 511vey, 1975;
McCu11a9h • Ne1der, 1989; Carr011 et a1., 1995).
D1REC7 EFFEC75 0N REPR0DUC710N
We w111 d15cu55 tw0 5119ht1y d1fferent v1ew5 0n the
d1rect effect 0n repr0duct10n rate, 0ne repre5ent5 an
effect 0n 5urv1va1 dur1n9 009ene515 and 0ne an effect
0n ener9et1c5.
Ha2ard m0de1
7he f1r5t v1ew 1nterpret5 effect5 0n repr0duct10n a5
a m0rta11ty dur1n9 009ene515. 7he 5en51t1ve per10d tH
15 a55umed t0 6e 5h0rt and f1xed. 7he t01erance
funct10n f0r repr0duct10n 4R(c, t) can then 6e wr1tten
a5
4R(C, t) = exp - {f + ~"/1~H(t,) d h } -~exp{ - tH/~¢.(t)}
(8)
where ,~¢H 15 the ha2ard rate f0r the 0vum,
wh1ch 15 taken t0 6e a 11near funct10n 0f the
c0ncentrat10n 0f t0x1c c0mp0und 1n the fema1e.
1t pr0ve5 t0 6e c0nven1ent t0 1ntr0duce the
•5tre55 funct10n• 5(c4) = t.//c. = tH1~,H(c4 - c0). =
c d ~ ( c 4 - c0)+, where /~,, 15 the k1111n9 rate dur1n9
009ene515 (K001jman, 1993; 8edaux • K001jman,
1994; K001jman • 8edaux, 1995). N0t1ce that the
k1111n9 rate re1ate5 t0 the t01erance c0ncentrat10n a5
/H/~+H ~- C1~ 1 .
7h15 effect 0n 5urv1va1 15 c0n515tent w1th the D E 8
the0ry f0r a91n9, wh1ch 5tate5 that the ha2ard rate 15
pr0p0rt10na1 t0 the c0ncentrat10n 0f accumu1ated
(6) d15funct10na1 pr0te1n5 1n the 60dy. 7he5e pr0te1n5 are
pr0duced 6y D N A that 15 affected 6y free rad1ca15
that are 1nherent t0 the re5p1rat10n pr0ce55. 7he
re5p1rat10n rate 15 r0u9h1y pr0p0rt10na1 t0 the u5e 0f
ener9y fr0m the re5erve5 (K001jman, 1995). 5ee
(7)
K001jman (1993) f0r a der1vat10n 0f the re1at10n5h1p
6etween a91n9 and ener9et1c5, the re1at10n5h1p w1th
effect5 0f muta9en1c c0mp0und5 and te5t5 a9a1n5t
exper1menta1 data. 7he ha2ard m0de1 f0r the
de5cr1pt10n 0f t0x1c effect5 0n repr0duct10n take5 the
 Daphn1a repr0duct10n t0x1c1ty te5t5
p05tu1ated 5h0rt 5en51t1ve per10d dur1n9 009ene515 a5
the 0n1y d1fference w1th effect5 0n 5urv1va1 0f the
repr0duc1n9 fema1e.
5u65t1tut10n 0f the ha2ard rate 1nt0 the repr0duc-
t10n rate 1ead5 t0
/~(c, t) =/~(0, t)exp{ -- 5(c4)}
= R(0, t)exp{ -- eft• -- c0)+} (9)
where e4 15 91ven 1n e4uat10n (5) and the parameter
cH w111 6e ca11ed the t01erance c0ncentrat10n (f0r
repr0duct10n).
C05t5 m0de1
7he 5ec0nd v1ew 0n d1rect effect5 0n the
repr0duct1ve 0utput 15 an 1ncrea5e 1n the ener9y c05t5
per e99; the 1atter 15 a9a1n taken t0 6e a 11near
funct10n 0f the c0ncentrat10n 0f t0x1c c0mp0und 1n
the an1ma1. Each m01ecu1e that exceed5 the hand11n9
capac1ty 0f the 1nd1v1dua1 ha5 the 5ame effect. 7h15
5tr1Ct v1ew can 6e re1axed 6y 5tat1n9 that the actua1
way 1n wh1ch the c0mp0und affect5 the ener9y c05t
per e99 15 c0mp1ex and n0n-11near, 6ut that we
appr0x1mate the de5cr1pt10n w1th tw0 term5 0f the
7ay10r expan510n ar0und the n0-effect c0ncentrat10n.
7h15 1ead5 t0 the 5tre55 funct10n 5(c4) = c£ ~(c4 -- c0)+,
where the parameter cR w111 6e ca11ed the t01erance
c0ncentrat10n (f0r effect5 0n the c05t5 per e99). 7he
re5u1t1n9 repr0duct10n rate 15
R(c, t) = R(0, t)(1 + 5(c4))-•
=/~(0, t)(1 + cR~(c4 -- c0)+)-• (10)
where e4 15 91ven 1n e4uat10n (5).
80th v1ew5 0n the d1rect effect5 0n repr0duct10n
1nv01ve the 5ame three parameter5: the n0-effect
c0ncentrat10n c0, the t01erance c0ncentrat10n cn, cR
and the e11m1nat10n rate/~a. 7he 1a5t parameter a150
0ccur5 1n the de5cr1pt10n 0f effect5 0n 5urv1va1 0f the
repr0duc1n9 fema1e. 7he tw0 repr0duct10n funct10n5
6ehave numer1ca11y very 51m11ar.
1ND1REC7 EFFEC75 0 N R E P R 0 D U C 7 1 0 N
7he effect5 0n ma1ntenance, 9r0wth and a551m11a-
t10n 1nd1rect1y affect repr0duct10n, 6y the c0n5erva-
t10n 0f ener9y pr1nc1p1e. 7h15 5ect10n eva1uate5 the5e
1nd1rect effect5 0n repr0duct10n, wh1ch w111n0t 0n1y
re5u1t 1n a decrea5e 0f the repr0duct10n rate, 6ut a150
1n a de1ay 0f the 0n5et 0f repr0duct10n. 7he
0ccurrence 0f 5uch a de1ay 15 the 6e5t 1nd1cat10n f0r
1nd1rect effect5.
Ma1ntenance
Many t0x1c c0mp0und5 are 11ke1y t0 affect
ma1ntenance re4u1rement5, rather than repr0duct10n
d1rect1y, wh1ch tran51ate5 1nt0 an 1ncrea5e 1n the
ma1ntenance c05t5. 8ecau5e ma1ntenance ha5 pr10r1ty
0ver 9r0wth 1n the DE8 the0ry, 5uch an 1ncrea5e
1ead5 t0 a reduct10n 0f the 9r0wth rate. 1f the an1ma1
w0u1d c0mpen5ate th15 reduct10n 0f 9r0wth 6y
1715
1ncrea51n9 the a110cat10n t0 ma1ntenance p1u5 9r0wth
and thu5 decrea51n9 the a110cat10n t0 repr0duct10n,
th15 w0u1d 6e 1nd15t1n9u15ha61efr0m the c05t5 m0de1
0f the d1rect effect 0n repr0duct10n, wh1ch ha5 ju5t
6een d15cu55ed. 7heref0re, we n0w a55ume that the
an1ma1 d0e5 n0t c0mpen5ate the reduct10n 0f 9r0wth.
51nce the feed1n9 rate depend5 0n 60dy 512e, the
feed1n9 rate 15 affected a5 we11. Repr0duct10n a150
depend5 0n 60dy 512e a5 we11 a5 0n feed1n9 rate (v1a
the effect 0n re5erve dynam1c5), 50 repr0duct10n 15
a150 affected, n0t 0n1y v1a a reduced a110cat10n t0
repr0duct10n, 6ut a150 v1a a de1ay 1n the 5tart 0f
repr0duct10n.
1n ana109Y w1th the d1rect effect5, we n0w a5-
5ume that the ma1ntenance rate c0eff1c1ent
15 rhc = tn0(1 + 5(c4)) w 1 t h 5tre55 funct10n
5(c4)=c~1(c4-c0)+, where cM 15 the t01erance
c0ncentrat10n f0r ma1ntenance and c0 the n0-effect
c0ncentrat10n; tn0 5tand5 f0r the ma1ntenance rate
c0eff1c1ent 1n the c0ntr01. 7he f0rmu1at10n 0f the
repr0duct10n rate d0e5 n0t acc0unt f0r effect5 0n the
ener9y c05t5 0f e995, 0n1y f0r the reduced ener9y
a110cat10n t0 repr0duct10n.
5u65t1tut10n 0f the effect 0n ma1ntenance 1nt0 the
9r0wth rate 0f e4uat10n (1), and the repr0duct10n rate
1n e4uat10n (3), 1ead5 t0
d
dt 1 = ~ ( f - - 1(1 + 5(c4))) (1 1)
k = (1 + 5(c.))-•- 1 ~ k{9~ -+- -1~ f 1 - - 1p~) (12)
where f 15 the v0n 8erta1anffy 9r0wth rate 1n the
c0ntr01 and 1 = (V/Vm) ~n 15 the 5ca1ed 1en9th. 7he
max1mum v01ume 1•m 1n the 5ca11n9 refer5 t0 the
max1mum v01ume 1n the c0ntr01. 7he c0up1ed
e4uat10n5 (11) and (5) t09ether de5cr16e 9r0wth and
the k1net1c5 0f the c0mp0und.
6r0wth
1f 9r0wth 15 affected, repr0duct10n w1116e affected
a5 we11, 6ecau5e feed1n9 depend5 0n 60dy 512e. 51nce
the 9r0wth rate d0e5 n0t affect the u1t1mate 512e, the
effect 15 a de1ay 1n repr0duct10n. 1n ana109y w1th the
d1rect effect5, we n0w a55ume that the ener9y
1nve5tment rat10 15 9c =90(1 +5(c4)) w1th 5tre55
funct10n 5(c4)---c~(c4--c0)+, where e~ 15 the
t01erance c0ncentrat10n f0r 9r0wth and c0 the
n0-effect c0ncentrat10n.
5u65t1tut10n 0f the effect 0n 9r0wth 1nt0 the 9r0wth
rate, e4uat10n (1), and the repr0duct10n rate,
e4uat10n (3), 1ead t0
d 1 = ~ ( f -- ~ f +9 (13)
dt 1 ) j + 9(1 + 5(c4))
km ((1+ 5(c0)9 +1f1, 10 (14)
/~ = 1 - 1~ •(1 + 5(c4))9 + f " -
1716 5. A. L. M. K001jman and J. J. M. 8edaux
A551m11at10n
1f a551m11at10n 15 affected, 1.e. the 1nc0m1n9 ener9y
15 reduced, repr0duct10n and 9r0wth are effected
a5 we11. 1n ana109y w1th the d1rect effect5, we
n0w a55ume that the ener9y 1nve5tment rat10
15 ~ = ~0(1 - 5(c4)) w1th 5tre55 funct10n
5(c4)=c£~(c4-c0)+, where cA 15 the t01erance
c0ncentrat10n f0r a551m11at10n and c0 the n0-effect
c0ncentrat10n. N0t1ce that the 5tre55 funct10n appear5
w1th a ne9at1ve 519n, rather than a p051t1ve 0ne, t0
m0de1 adver5e effect5 0n a551m11at10n. 7he c0n5e-
4uence 15 that c4 < cA + c0 mu5t h01d, 50 a150
c < cA + c0 f0r a11 ch05en te5t c0ncentrat10n5 c. 7he
c0n5tra1nt 0n the va1ue f0r cA 15 1n fact a 61t 5tr0n9er
than th15, 6ecau5e the a551m11at10n rate mu5t exceed
the ma1ntenance re4u1rement5. 7he D E 8 the0ry
5tate5 that the 1nd1v1dua1 d1e5 6y 5tarvat10n 1f 1t 15
una61e t0 m061112e en0u9h ener9y fr0m 1t5 re5erve5
f0r ma1ntenance purp05e5.
5u65t1tut10n 0f the effect 0n a551m11at10n 1nt0 the
9r0wth rate e4uat10n (1) and the repr0duct10n rate
e4uat10n (3) 1ead5 t0
d t =1 ,/(f(1 - 5(c4)) - 1)
7he tw0 m0de15 f0r d1rect effect5 0n repr0duct10n
and the three 1nd1rect 0ne5 are 111u5trated a5 re5p0n5e
5urface5 a60ve the exp05ure t1me-c0ncentrat10n p1ane
1n F19. 4.
5tat15t1c5
7he tran51at10n 0f the repr0duct1ve eff0rt 1nt0 the
num6er 0f 0ff5pr1n9 1nv01ve5 the m0u1t1n9 cyc1e and
deta115 0f the exper1menta1 pr0t0c01. 7he te5t med1a
are u5ua11y renewed three t1me5 a week and the
num6er 0f 5urv1v1n9 parent5 and 0ff5pr1n9 are
c0unted 0n a da11y 6a515. 7he 1nterm0u1t per10d 15 a
61t f1ex161e and the an1ma1 tend5 t0 5ynchr0n12e 1t5
m0u1t1n9 cyc1e w1th the renewa1 cyc1e; 500n after the
chan9e 0f te5t med1um 1t de11ver5 and m0u1t5. When
c0unt1n9 da11y at 20°C, m0u1t5, and thu5 y0un9, w111
6e 065erved 0n1y three t1me5 a week. We n0w f0cu5
0n th05e 065ervat10n5 at t1me p01nt5 t~*, where a
m0u1t ha5 6een pre5ent. (1f a m0u1t 15 pre5ent, 1t 15
11ke1y t0 6e pre5ent f0r a1m05t a day 6ecau5e m0u1t1n9
0ccur5 ju5t after renewa1 0f the med1um and c0unt1n9;
6ut we w111 ne91ect th15 deta11.) 7he expected num6er
0f 0ff5pr1n9 0f a part1cu1ar fema1e at exp05ure t1me
t~* 1n c0ncentrat10n c1 e4ua15
N(t,*, cj) = 4R(cj, t1*-1) R(0, t) dt
2
~- 4R(c, t,*~)(t,*~, - t~*2)k(0, t~ ~) (17)
7h15 f0rmu1at10n d0e5 ju5t1ce t0 the 6101091ca1 fact
that the a110cat10n t0 repr0duct10n take5 p1ace dur1n9
the 1nterm0u1t per10d pr10r t0 the 0ne that de11ver5
the hatch11n95.
7he m0u1t5 are u5ua11y n0t 065erved, 50 that th15
ana1y515 15 perhap5 t00 deta11ed f0r pract1ca1
purp05e5. We m19ht try t0 ne91ect the m0u1t1n9 cyc1e
and ju5t f1t the num6er 0f 0ff5pr1n9 a5 1f the
repr0duct10n pr0ce55 15 c0nt1nu0u5. 7he fre4uent
2er05 c0rrect the 6r00d5 f0r the r19ht mean, 6ut the
var1ance 151ar9e, 0f c0ur5e. 7h15 91ve5 pr061em51n the
parameter e5t1mat10n. A1ternat1ve1y, we m19ht f1t the
cumu1at1ve repr0duct10n. 7h15 reduce5 the e5t1mat10n
pr061em, 6ut the c0unt5 are n0w 5tat15t1ca11y
dependent. Numer1ca1 5tud1e5 1nd1cate that th15
hard1y affect5 p01nt e5t1mate5.
7he 065erved num6er 0f 0ff5pr1n9 15 emp1r1ca11y
a55umed t0 f0110w the n0rma1 d15tr16ut10n appr0x1-
mate1y, w1th a f1xed c0eff1c1ent 0f var1at10n. 7h15
c0eff1c1ent 5h0u1d 6e 1e55 than 0.25 f0r the t0x1c1ty te5t
t0 6e accepta61e acc0rd1n9 t0 the 0 E C D 9u1de11ne.
7he e5t1mat10n 0f the parameter va1ue5 u51n9 the
max1mum 11ke11h00d meth0d can 6e 51mp11f1ed a5 a
f1r5t appr0x1mat10n 6y a 1ea5t 54uare5 meth0d w1th
we19ht c0eff1c1ent5 1nver5e1y pr0p0rt10na1 t0 1 p1u5
(15) the 54uared num6er 0f 065erved 0ff5pr1n9 (McCu1-
1a9h • Ne1der, 1989; Carr011 et a1., 1995). N0t1ce that
the e5t1mat10n 0f the n0-effect c0ncentrat10n c0 and
the e11m1nat10n rate/~a depend 0n the type 0f effect 0n
repr0duct10n. 7he 5ca1ed 1en9th5, 5ca1ed t155ue-c0n-
centrat10n5 and the cumu1at1ve repr0duct1ve 0utput
ha5 t0 6e 06ta1ned numer1ca11y, u51n9 e.9. Adam5
pred1ct0r-c0rrect0r meth0d (8urden • Fa1re5, 1985).
1f effect5 0n repr0duct10n and 5urv1va1 0ccur at the
5ame t1me, the 065ervat10n5 5h0u1d 6e ana1y2ed
51mu1tane0u51y, 6ecau5e 60th de5cr1pt10n5 have the
5ame e11m1nat10n rate.
REDUCED M 0 D E L 5
Many 0r9an1c c0mp0und5 have an e11m1nat10n rate
that 15 1ar9e w1th re5pect t0 the v0n 8erta1anffy
9r0wth rate (Hawker • C0nne11, 1986), 50 that f0r
/¢~>>); e4uat10n (5) reduce5 t0 c4 = c. 7he 5urv1va1
pr06a6111ty and repr0duct10n rate reduce t0
.~(c, t) = ~ ( 0 , t) exp{ - t/¢+(c - c0)+ } (18)
R(c, t) = R(0, t) exp{ - cH ~(c - c~,)+ ] (19)
= /~(0, t)(1 + cR•(c - c0)+) ~ (20)
7he 5tre55 funct10n5 f0r the 1nd1rect effect5 0n
repr0duct10n are c0n5tant5, 6ut the repr0duct10n rate
5t111 ha5 t0 6e 06ta1ned numer1ca11y.
F0r the chr0n1c Daphn1a te5t we can hard1y ne91ect
the chan9e 1n 60dy 512e dur1n9 exp05ure, 6ut f0r a
num6er 0f 0ther t0x1c1ty te5t5 we can. F0r 1 =.1~ the
5ca1ed t155ue-c0ncentrat10n e4uat10n (5) reduce5 t0
¢ 4 ( t ) = c ( 1 - e x p { - t/¢,}), and the 5urv1va1 pr06-
a6111ty and repr0duct10n rate 6ec0me5 f0r c > c0 and
t>t0= -/¢5~1n{1-c0/c]
 Daphn1arepr0duct10n t0x1c1ty te5t5 1717

ha2ard, c~ = 1.62 #9 1-1 ma1nt., cM -" 6.02 #9 1-1

~[
0
•7

•
t20

W
4.

0 46

0 ~
e

12 16
C0Pc° 28 U 90r1¢•
*¢ 1 ~ e ~ d ~ 1r0e, C1

c05t5, cn -- 1.35 #9 1-1 9r0wth, e~ = 0.61 #9 1-1

; t29

,• .e

0
0 0
¢

~ 12 18
u c0 c" 28 u C00c*
t1mQ, C~

a551m., cA = 9.57 #9 1-1

; t~

r
e M

C0~9 ~ 12 15 2~
t 1 r119 ~ C1

F19. 4. D1rect (1eft c01umn) and 1nd1rect (r19ht c01umn) effect5 0n repr0duct10n. 7 h e var10u5 t01erance
c0ncentrat10n are ch05en 50 that the effect 512e 15 51m11ar. 7 h e e11m1nat10n rate 15 5et e4ua1 t0 the v0n
8erta1anffy 9r0wth rate, /~ = ~ = 0.1 d •, and the n0-effect c0ncentrat10n 15 c0 = 1.5 tt9 1-•].
1718 5. A. L. M. K001jman and J. J. M. 8edaux

/~9 C d 1- ~ /29 C d 1-~

100
ha2ard /. 0 c05t5 /. 0

~04
E
/0.2 = .e
Q 60 0 60
), ),

0
40

20 • 3 20
u
1
2 e eee 1e e De e • ~ . . . .

4 8 12 16 20 4 0 ~2 ~16 20
t1me, d t1me, d

F19. 5.7he mean cumu1ated num6er 0f y0un9 per fema1e Daphn1d a5 a funct10n 0f the exp05ure t1me t0
5evera1 c0ncentrat10n5 0f cadm1um. 7he f1tted curve5 repre5ent 1ea5t 54uare5 f1t5 0f the ha2ard (1eft) and
the c05t (r19ht) m0de1 f0r effect5 0n repr0duct10n. 61ven an e11m1nat10n rate 0f/~ = 0.05 d-~, the e5t1mated
parameter5 (and 5tandard dev1at10n5) are c0 = 0.023(0.004) p91-~, c~ = 0.166(0.006) #91 ~,
/1~m : 13.1(0.2) d-~ and c0 = 0.047(0.002) p9 1 ~, cR = 0.069(0.003) #9 1-~, / ~ = 13.1(0.2) d ~ re5pect1ve1y.

~ ( c , t) = ~ ( 0 , t)exp{/~,/~£ •c(exp{ -- t0/~a} e5t1mated 1n e4uat10n5 (24) and (26). 1t w111 6e hard
t0 1dent1fy 1nd1rect effect5 0n repr0duct10n 1f the
-exp{ - - t]~a}) - ]~,(c -- c0)(t - t0)} (21) exp05ure t1me 15 5h0rt re1at1ve t0 the 1nver5e 0f the
/~a5ma11 v0n 8erta1anffy 9r0wth rate.
= ~ ( 0 , t)exp{ - (t - t0)2k~c} (22)

R(c, t) = R(0, t) 7E575 A6A1N57 EXPER1MEN7AL DA7A

×exp{ - cH~(c(1 - exp{ - t/~.}) - c0)+ }
/~a5ma11
= /~(0, t)exp{ - c f f •(Ctff5a -- C0)+ }
/~(c, t) =/~(0, t)
× (1 + cR 1(C(1 -- exp{ - /]~a}) -
/~a5ma11
= /~(0, t)(1 + c ~ ( c t f : a -- c0)+)-~
7 h e der1vat10n 0f e4uat10n (22) 15 1e55 5tra19htf0r-
ward, 6ecau5e 0ur 5trate9y t0 dea1 w1th env1r0nment-
c0ncentrat10n5, rather than t155ue-c0ncentrat10n5
w0rk5 0ut a 61t m0re c0mp1ex here. We have t0
c0n51der the 11m1t/~a ~ 0, wh11e the 610c0ncentrat10n
c0eff1c1ent Pv0--* ~ , 5UCh that the Uptake rate
[/~d,] = Pva/~a rema1n5 f1xed, and the k1111n9 rate
/~, ~ m, 5UCh that the •k1111n9 aCCe1erat10n• k~t =/~d~a
rema1n5 f1Xed a5 We11. 7he t155Ue-C0nCentrat10n
am0Unt5 t0 [ Q ] ( t ) = [1~d~]ct and the ha2ard rate t0
1~ = k~tC(t -- t0). 7he t1me parameter t0 1n e4Uat10n
(22) ha5 the 1nterpretat10n t0 = [Q]0([/~d,]c)-~, Where
[Q]0 5tand5 f0r the n0-effeCt-t155Ue-C0nCentrat10n.
7h15 t1me parameter 15thU51nVer5e1y pr0p0rt10na1 t0 C.
N0t1Ce that the de5Cr1pt10n 0f the effect5 have 0n1y
tW0 parameter5 1f the e11m1nat10n rate 15 5ma11,
6ecau5e 0n1y k~/cn, ~a/CR and C0/CR, C0/CR can 6e
We 91ve 0ne examp1e 0f a d1rect effect and tw0 0f
1nd1rect effect5, a11 taken fr0m a r1n9 te5t that wa5
(23)
0r9an12ed 6y the 0 E C D . We u5ed the cumu1at1ve
num6er 0f y0un9 per fema1e, t0 fac111tate the
(24)
e5t1mat10n 0f parameter5 and t0 1mpr0ve the
pre5entat10n 0f the re5u1t5. 7he 5tandard dev1at10n5
are meant t0 6e 1nd1cat1ve. An ana1y515 6a5ed 0n
c 0 ) + ) -1 m0u1t1n9 cyc1e5 wa5 n0t p055161e due t0 1ack 0f
(25) 065ervat10n5 0f the m0u1t5.
F19ure 5 5h0w5 the effect5 0f cadm1um 0n the
(26) repr0duct10n 0f D. ma9na, w1th the 1ea5t 54uare5 f1t
0f the ha2ard and the c05t m0de1. 80th m0de15 f1t
we11, the ha2ard m0de1 perhap5 5119ht1y 6etter. 7he
n0-effect c0ncentrat10n5 0f 60th m0de15 d1ffer 6y a
fact0r 0ftw0, 6ut 1n v1ew 0fthe fact that they repre5ent
ju5t 11% and 23% 0 f t h e 10we5t te5ted c0ncentrat10n,
the accuracy 5eem5 4u1te accepta61e. 7he e11m1nat10n
rate c0u1d n0t 6e e5t1mated fr0m the5e data. 51nce n0
cadm1um-1nduced de1ay 0f the 0n5et 0f repr0duct10n
ha5 6een 065erved, we have n0 1nd1cat10n5 f0r
1nd1rect effect5 0f cadm1um 0n repr0duct10n.
7he 5ec0nd examp1e (F19. 6) 5h0w5 the effect5 0f
phen01 0n the repr0duct10n 0f D. ma9na, w1th the
1ea5t 54uare5 f1t 0f the ma1ntenance, 9r0wth and
a551m11at10n m0de1. D1rect effect5 can 6e exc1uded 0n
the 6a515 0f a phen01-1nduced de1ay 0f the 0n5et 0f
repr0duct10n. 7he three m0de15 f1t we11; the mean
dev1at10n 6etween the 065erved num6er 0f y0un9 and
m0de1 expectat10n5 are 5.7, 5.4 and 5.3 f0r effect5 0n
 Daphn1a repr0duct10n t0x1c1ty te5t5 1719

ma1ntenance, 9r0wth and a551m11at10n, re5pect1ve1y. 57ANDARD 57A71571CAL ANALY5E5

7he n0-effect c0ncentrat10n hard1y depend5 0n
the type 0f effect5. 7he e11m1nat10n rate c0u1d n0t 7he 5tat15t1ca1 ana1y515 0f 5urv1va1 data 15
6e e5t1mated fr0m the5e data. 7he 0n5et 0f 5tandard12ed 6etter than that 0f repr0duct10n data.
repr0duct10n 1n the c0ntr01 15 a 61t 1ater than u5ua1, Fre4uent1y app11ed 5urv1va1 and repr0duct10n m0de15
wh1ch ha5 6een taken 1nt0 acc0unt 6y ch0051n9 are 6a5ed 0n the 109-10915t1c funct10n
2p = 0.55.
7he th1rd examp1e (F19. 7) 5h0w5 the 5ame f0r
3,4-d1ch10r0an111ne. 7 h e de1ay 0f the 0n5et 0f 4(c, t) = 1 + ~,cL50(t)) ] (27)
repr0duct10n p01nt5 t0 an 1nd1rect effect. 7he three
m0de15 f1t we11; the mean dev1at10n 6etween the
065erved num6er 0f y0un9 and m0de1 expectat10n5
are 5.0, 4.6 and 4.7 f0r effect5 0n ma1ntenance, (28)
9r0wth and a551m11at10n, re5pect1ve1y.

m9 phen011-1 m9 phen011-1

,20 0, 0.32, 0.56

~ ma1ntenance r0
• •, 0.56 •

a0 ." ~ a0 ." *•

~ ~ 1.8
,_
600 . , ~-
J °01~
# ¢ / / ~ / 0
0 ,0 :;.. . 0 ,0 I "•
u 20 • 1 1 u 20
,, ~, , 3 . 2 ~ ..3.2

• 0 ~ ~ - - - " ° ° °

0 4 8 12 16 20 0 4 8 12 10 28

t1me, d t1me, d

m9 phen011-1

"11 a551m11at1°n
~
•
~
,~e

~0 0
.~ 0, 0.32
0.56

~- 60
." *1.8

0
0 ~0 " • 3.2
• . ee e

0
e 4 • 12 16 28

t1me, c1

F19. 6.7he mean cumu1ated num6er 0f y0un9 per fema1e Daphn1da5 a funct10n 0f the exp05ure t1me t0
5evera1 c0ncentrat10n5 0f phen01. 7he f1tted curve5 repre5ent 1ea5t 54uare5 f1t5 0f the ma1ntenance (upper
1eft), the 9r0wth (a60ve) and the a551m11at10n (1eft) m0de1 f0r effect5 0n repr0duct10n. 61ven an e11m1nat10n
rate 0f/~ = 0.5 d •, the e5t1mated parameter5 (and 5tandard dev1at10n5) are

c0. m91-• c,, m91-~ Rm, d •

ma1nt. 0.41 (0.04) 6.23 (0.36) 18.19 (0.30)
9r0wth 0.60 (0.04) 0.73 (0.06) 17.57 (0.22)
a551m. 0.37 (0.05) 11.34 (0.58) 18.23 (0.28)

WR 3 0 / 7 ~
1720 5. A. L. M. K001jman and J. J. M. 8edaux

#9 DCA1- t # 9 DcA •-1

0, 3.2 0, 3.2
7
ma1ntenance 5.6 7
9r0wth ~ 5.6

68 5e

• •0 c~
• 1

10 ~" 48
• 10
40
0
0
0 • • E 11 0•0

• •• 0•• 18 ~ 28
• •• • 0 18
e8 D
u ~•0.,, u • 600 0
32 "" 32

8 8
8 4 8 12 16 28
e A 8 12 16 20
t1me, d
t1me, d

#9 DCA 1-1

0, 3.2
7
a551m11at10n / ~ 5.6
68

0
40
" . •• 10
00
0

28 • • " 0e0 18
u
1 00~, 1*
" 32

4 8 12 16 20

t1me, d

F19. 7.7he mean cumu1ated num6er 0f y0un9 per fema1e Daphn1d a5 a funct10n 0f the exp05ure t1me t0
5evera1 c0ncentrat10n5 0f 3,4-d1ch10r0an111ne. 7he f1tted curve5 repre5ent 1ea5t 54uare5 f1t5 0f the m0de1
f0r effect5 0n repr0duct10n v1a ma1ntenance, 9r0wth and a551m11at10n. 7he e5t1mated parameter5 are

c0, P91 1 c.,#91 ~ ~,, d-~ /~m,d n

ma1m. 3.84 (1.24) 31.7 (5.6) 0.75 (1.75) 9.50 (0.21)
9r0wth 3.51 (2.55) 3.85 (1.4) 0.90 (3.58) 9.56 (0.19)
a551rn. 3.37 (1.22) 65.8 (12.5) 0.87 (2.15) 9.53 (0.20)

where cL50den0te5 the LC50, 1.e. the c0ncentrat10n at wh1ch 15 the h19he5t te5ted c0ncentrat10n that d0e5
wh1ch the 5urv1va1 pr06a6111ty 15 ha1f that 1n the n0t d1ffer 519n1f1cant1y fr0m the c0ntr01. Exten510n5 0f
c0ntr01, f1 15 the 9rad1ent 0f the c0ncentrat10n-re- the 5tandard 109-10915t1c m0de1 t0 1nc1ude a n0-effect
5p0n5e funct10n, N ( c ) = ~0~/~(c, t ) d t 15 the expected c0ncentrat10n a5 a parameter have a150 6een
cumu1at1ve num6er 0f 0ff5pr1n9 at 21 d, cE50 15 pr0p05ed (K001jman, 1981). 7he m0t1vat10n wa5 t0
the 50% effect c0ncentrat10n (u5ua11y n0tated a5 501ve the pr061em 0f mak1n9 an err0r 0f the 5ec0nd
EC50) and f1R 15 the 9rad1ent parameter f0r k1nd 6y n0t reject1n9 the nu11-hyp0the515 that the
repr0duct10n. K001jman (1981) t1ed the cL~0(t) c0mp0und 1n the te5ted c0ncentrat10n ha5 n0 effect,
parameter t0 the t155ue-c0ncentrat10n, wh1ch 1ead5 wh11e 1n fact 1t d0e5 have an effect. 7he c0n5e4uence
f0r a 0ne-c0mpartment uptake/e11m1nat10n k1net1c5 0f 5uch an err0r 15 that 510ppy te5t pr0cedure5 1ead t0
t0 cL50(t) = cL50~(1 - exp{ - t/~a}) 1. 7he 5urv1va1 h19h n0-effect c0ncentrat10n5, a m05t unf0rtunate
pr06a6111ty 1n the c0ntr01 15 de5cr16ed 6y c0up11n9 0f pr0pert1e5.
~ ( 0 , t) = exp{ - t/~0}. 7he e5t1mat10n 0f LC50 7he parameter5 t0 6e e5t1mated are/~0,/~, cL50~, f1
va1ue5 ha5 6een d15cu55ed 6y H0ek5tra (1993). and c0 f0r 5urv1va1 and N(0), cE50, f1R and c0 f0r
1ndependent fr0m and 1nc0n515tent w1th the5e repr0duct10n.
m0de15, the n0-065erved-effect c0ncentrat10n 7he5e 5tandard m0de15 are pr1mar11y de5cr1pt1ve,
( N 0 E C ) f0r 5urv1va1 and repr0duct10n 15 e5t1mated, rather than mechan15t1c. K001jman (1993) 91ve5
 Daphn1arepr0duct10n t0x1c1tyte5t5
ar9ument5 why 1nterpretat10n5 0f the 109-10915t1c
re5p0n5e m0de15 1n term5 0f 5catter d15tr16ut10n5 0f
thre5h01d va1ue5 are weak f0r effect5 0n 5urv1va1. 7he
ar9ument5 d0 n0t 5eem t0 app1y t0 effect5 0n
repr0duct10n, 6ecau5e we can ea511y 065erve that the
repr0duct10n rate 15 affected f0r each 1nd1v1dua1.
Effect5 0n repr0duct10n 0f a c0h0rt 0f 1nd1v1dua15
cann0t 6e under5t00d 1n term5 0f affected and
unaffected 1nd1v1dua15, wh1ch 15 1nherent t0 the
c0ncept 0f thre5h01d va1ue.
7he 9rad1ent parameter5 d0 n0t have a u5efu1
funct10n. 1n fact, they c0mp11cate the e5t1mat10n 0f
the n0-effect c0ncentrat10n5 c0n51dera61y, 6ecau5e a
free 9rad1ent parameter reduce5 t0 1nf0rmat10n
c0ntent 0f an 065erved effect at a part1cu1ar
c0ncentrat10n t0 ••the n0-effect c0ncentrat10n mu5t 6e
10wer than th15 part1cu1ar c0ncentrat10n••. 1f the
de5cr1pt10n 0f effect5 d0e5 n0t 1nc1ude a free 9rad1ent
parameter, 5uch a5 1n th15 art1c1e, 065erved effect5 d0
c0nta1n 1nf0rmat10n a60ut the va1ue 0f the n0-effect
c0ncentrat10n. 7he u5e 0f the 9rad1ent parameter t0
ca1cu1ate 5ma11-effect c0ncentrat10n5 15 0f 11m1ted
va1ue, 6ecau5e 5uch va1ue5 depend 5en51t1ve1y0n the
deta11ed 5hape 0f the ta11 0f the 519m01d re5p0n5e
funct10n. 7h15 15 pr061emat1c 6ecau5e a 519m01d
re5p0n5e funct10n 0n1y ha5 an emp1r1ca1 6a515, w1th
hard1y any 5upp0rt f0r the deta11ed 5hape 0f the ta115.
1t 15 a1m05t 1mp055161e t0 ju5t1fy a part1cu1ar ch01ce
emp1r1ca11y. Pract1ce teache5 that 1t 15 p055161e t0
e5t1mate a p051t1ve n0-effect c0ncentrat10n a5 a
parameter 6u11t 1nt0 the 109-10915t1c m0de1 1n a60ut
ha1f 0f the re5u1t5 fr0m typ1ca1 t0x1c1ty te5t5, and 1n
a60ut 0ne 0ut 0f ten ca5e5, 1t d1ffer5 5tat15t1ca11yfr0m
2er0. C0n5e4uent1y, 1t 15 0f a 11m1ted pract1ca1 va1ue.
D15CU5510N AND C0NCLU510N5
We a55umed that the f00d den51ty 15 c0n5tant, 50
that the re5erve den51ty 15 a150 c0n5tant. 7h15, 0f
c0ur5e, 0n1y h01d5 1f the an1ma1 15 "1n e4u1116r1um••
w1th th15 f00d ava11a6111ty. 1f f00d den51ty chan9e5, 0r
1f there 15 n0 900d at a11, we have t0 acc0unt f0r the
chan9e 1n 11p1d c0ntent 0f the an1ma1, 6ecau5e the
uptake/e11m1nat10n 6ehav10ur can 6e rather 5en51t1ve
f0r 5uch chan9e5. We a150 need re5erve5 t0 m0de1
e11m1nat10n v1a repr0duct10n, 51nce fre5h1y pr0duced
e995 c0n515t a1m05t exc1u51ve1y0f e995.7he deta115 f0r
effect5 0f chan9e5 1n 11p1d c0ntent have 6een w0rked
0ut (K001jman • van Haren, 1990; K001jman, 1993).
8ecau5e the pre5ent de5cr1pt10n f0r effect5 0f c0m-
p0und5 15 0n the 6a515 0f the t155ue-c0ncentrat10n5,
var1at10n5 1n t1me can 6e taken 1nt0 acc0unt. 5uch
var1at10n5 1nc1ude meta6011c tran5f0rmat10n 0f the
c0mp0und.
50met1me5 a 5t1mu1at10n 0f repr0duct10n 15
065erved at 10w c0ncentrat10n 0f c0mp0und5 that
reduce repr0duct10n at h19her c0ncentrat10n5. 7h15
p00r1y under5t00d phen0men0n 15 ca11ed h0rme515.
50met1me5 1t 15 pre5ent when f00d 15 5upp11ed 1n 1ar9e
am0unt5, 6ut a65ent when f00d 15 5upp11ed 1n
1721
m0derate am0unt5. 7h15 1nd1cate5 that 5ec0ndary
5tre55 15 1nv01ved, at 1ea5t 1n 50me ca5e5. 0 n e 50urce
m19ht 6e p01y5acchar1de5 that 50me 9reen a19a
(Ch10re11a) pr0duce when they 0ccur 1n h19h den51t1e5
and/0r 6acter1a that dec0mp05e ce115 1n p00r
c0nd1t10n5 (a19ae are u5ua11y kept 1n 5t0ck 1n a
c0ncentrated f0rm f0r 50me t1me, wh1ch d0e5 n0 f00d
f0r the1r c0nd1t10n, and te5t med1a are 1e555u1ta61e f0r
a19a1 9r0wth). 8ecau5e 0f the5e uncerta1nt1e5,
h0rme515 15 n0t 1nc0rp0rated 1n the m0de1 ana1y515
and we 6e11eve that 1t can 6e av01ded 6y 0pt1m121n9
te5t c0nd1t10n5, at 1ea5t 1n 50me ca5e5.
7he advanta9e5 0f the pr0p05ed ana1y515c0mpared
w1th the 5tandard meth0d are:
• 7he 6a515 15 much m0re mechan15t1c, wh1ch
fac111tate5
- - t h e 11nk w1th phy51c0-chem1ca1 pr0pert1e5 0f
the c0mp0und5 (e.9. 11p0ph111c1ty)
- - t h e c0mpar150n 0f data fr0m 0ther t0x1c1ty
te5t5 (5uch a5 the 5urv1va1/9r0wth te5t 0f f15h
1arvae), effect5 0f d1fference5 1n exp05ure t1me5,
60dy 512e, etc.
- - t h e c0n5truct10n 0f a 5er1e5 0f c0n515tent
m0de15, ran91n9 fr0m 51mp1e 0ne5 f0r r0ut1ne
app11cat10n5, t0 advanced 0ne5 f0r re5earch
purp05e5.1t 15, f0r 1n5tance, 5tra19htf0rward h0w
t0 dea1 w1th c0mp0und5 that d15appear fr0m the
med1um, a5 a re5u1t 0f de9radat10n. We f1r5t have
t0 m0de1 the d15appearance pr0ce55, 6y a f1r5t
0rder pr0ce55 f0r 1n5tance, and 1n5ert the re5u1t
1n e4uat10n (5). 7h1515 a11 we have t0 d0 t0 arr1ve
at the c0rrect effect-character15t1c5.
- - t h e 1nc0rp0rat10n 0f add1t10na1 1nf0rmat10n,
5uch a5 the 60dy 1en9th at 21 d. 5uch extra
1nf0rmat10n he1p5 t0 1dent1fy the m0de 0f act10n
0f the c0mp0und (50 t0 ch005e 6etween the
1nd1rect effect m0de15) and t0 reduce the
c0nf1dence 1nterva15 f0r the parameter5 that have
t0 6e e5t1mated.
• 7he m0de15 d0 n0t have 9rad1ent parameter5.
7h15 501ve5 the e5t1mat10n pr061em5 0f the
n0-effect c0ncentrat10n5 (5ee 5ect10n 57AN-
DARD 57A71571CAL ANALY515). 7he5e
n0-effect c0ncentrat10n5 are n0w 1nte9rated 1n
the m0de1 a5 a parameter that can a55ume va1ue5
f0r c0ncentrat10n5 that have n0t 6een te5ted (a5
0pp05ed t0 N0EC•5). 7he reduct10n 0f the
num6er 0f parameter5 that have t0 6e e5t1mated
51mp11f1e5 the e5t1mat10n pr0cedure and reduce5
the c0nf1dence 1nterva15.
• 7he parameter5 (n0-effect c0ncentrat10n, k1111n9
rate and t01erance c0ncentrat10n) are 1ndepen-
dent fr0m the exp05ure t1me. N0t1ce that the
5tandard parameter5 (N0EC, LC50, EC50,
9rad1ent5) d0 depend 0n the exp05ure t1me,
wh1ch reduce5 the u5efu1ne55 1n r15k a55e55ment
and the c0mpar150n w1th 0ther t0x1c1ty data.
• 7he pre5ent character12at10n 0f the effect5 0n
1722 5. A. L. M. K001jman and J. J. M. 8edaux
5urv1va1 and repr0duct10n 15 fu11y pr0ce55
0r1ented, wh1ch mean5 that
- - e f f e c t 5 are ana1y2ed a5 a re5p0n5e 5urface
a60ve the exp05ure t1me-c0ncentrat10n p1ane.
7h15 a110w5 the ca1cu1at10n 0f 0ther character-
15t1c5, 5uch a5 the EC50 and 9rad1ent va1ue5.
--effect5 can 6e tran51ated 1nt0 effect5 0n the
p0pu1at10n 9r0wth rate. 7h15 15 n0t p055161e f0r
the 5tandard meth0d, 6ecau5e repr0duct10n 15
n0t treated a5 a pr0ce55, 50 that a de1ay 1n
repr0duct10n cann0t 6e d15t1n9u15hed fr0m a
reduct10n 0f the repr0duct10n rate.
A1th0u9h the var10u5 pathway5 t0 affect repr0duc-
t10n m19ht 6e hard t0 d15t1n9u15h fr0m repr0duct10n
data, the c0n5e4uence5 f0r the p0pu1at10n 1eve1 can
d1ffer c0n51dera61y (K001jman • Met2, 1983). 7he
rea50n 15 that effect5 0n ma1ntenance are fe1t 5tr0n91Y
when the p0pu1at10n 15 at 1t5 carry1n9 capac1ty and
a1m05t a11 0f the ener9y 6ud9et 15 5pent 0n
ma1ntenance, 6ut when the p0pu1at10n 15 1n a
pr0duct1ve pha5e, the5e effect5 are hard1y fe1t 6ecau5e
ma1ntenance take5 0n1y a 5ma11 part 0f the ener9y
6ud9et.
7he p0pu1at10n 9r0wth rate/1 can 6e 06ta1ned fr0m
the 5urv1va1 pr06a6111ty ~ ( a ) at a9e a and the
repr0duct10n rate/~(a) 1n c0n5tant env1r0nment5 v1a
the character15t1c e4uat10n
x 5tandard 0ne, 1t 15 p055161e t0 06ta1n the 5tandard
1=
1 exp{ -- ~t}~(t)R(t) dt
7he a91n9 pr0ce55 can 6ec0me 1mp0rtant f0r the
5urv1va1 pr06a6111ty at h19h a9e5 (5ee the 5ect10n 0n
the ha2ard m0de1).
1f the env1r0nment 15 n0t c0n5tant, we have t0 fa11
6ack t0 the the0ry 0f 5tructured p0pu1at10n dynam1c5
t0 eva1uate the p0pu1at10n c0n5e4uence5 0f the effect5
0f c0mp0und5 0n 5urv1va1 and repr0duct10n. 5ee
Met2 • D1ekmann (1986) f0r an 1ntr0duct10n.
7he c0mpar150n 0f the t0x1c1ty 0f c0mp0und5 w1th
d1fferent effect5 0n repr0duct10n 15 rather c0mp1ex.
0 n e p05516111ty 15 t0 c0mpare the p0pu1at10n 9r0wth
rate a5 a funct10n 0f the c0ncentrat10n, a5 06ta1ned
fr0m 1 = 50" exp{ - / J t } / ~ ( t ) dt. 7h15 ha5 the advan-
ta9e that de1ay51n the 0n5et 0f repr0duct10n are taken
1nt0 acc0unt and that 1t 11nk5 up 6etter w1th the
tran51at10n 0f 065erved effect5 0n 1nd1v1dua1 repr0-
duct10n 1n the 1a60rat0ry t0 p0pu1at10n c0n5e4uence5
0utd00r5. F19ure 8 111u5trate5 h0w the p0pu1at10n
9r0wth rate depend5 0n the env1r0nment-c0ncen-
trat10n f0r the var10u5 m0de5 0f act10n 0f the
c0mp0und.
7he pre5ent de5cr1pt10n re4u1re5 the app11cat10n 0f
rather advanced numer1ca1 techn14ue5.7h15 pract1ca1
hand1cap can 6e e11m1nated 6y app11cat10n 0f
50ftware packa9e D E 8 t 0 x a5 pr0v1ded 1n K001jman
• 8edaux (1996). 7he rap1d 1ncrea5e 0f the
perf0rmance 0f 10w-c05t per50na1 c0mputer5 m1n1-
m12e5 th15 pr061em 0f app11cat10n.
0.8
0.6 .~" C05t5
9r0wth
0.4 ma1nt.
ha2ard
002
c0nc./c0 • .
0 , , , , , , , , ~ , a551m.
0 2 4 6 10
F19. 8. 7he p0pu1at10n 9r0wth rate a5 a funct10n 0f the
env1r0nment-c0ncentrat10n f0r c0mp0und5 that have d1rect
and 1nd1rect effect5 0n repr0duct10n. 7he parameter va1ue5
are the 5ame a5 1n F19. 3.
7he ex15tence 0f 1ar9e data 6a5e5 0f LC50/EC50
va1ue5 w111 d0u6t1e551y de1ay the 1mp1ementat10n 0f
any 1mpr0vement 0n the 5tandard de5cr1pt10n 0f
effect5. F0r c0mpat16111ty rea50n5 w1th the5e data
6a5e5, 1t m19ht 6e 1ntere5t1n9 t0 ca1cu1ate LC50 and
EC50 t1me curve5 fr0m the pre5ent de5cr1pt10n. 7h15
can 6e d0ne 6y numer1ca11y 501v1n9 the e4uat10n5
4(cL50, t) = • and /~(cE50, t) = /~(0, t)/2. A1th0u9h the
pre5ent de5cr1pt10n ha5 1e55 parameter5 than the
parameter5 fr0m the pre5ent 0ne, 6ut 1t 15 n0t p055161e
t0 06ta1n the new parameter5 fr0m the 5tandard 0ne5.
7he 0r191na1 5urv1va1/repr0duct10n data are re4u1red
t0 06ta1n them.
We c0nc1ude that the pre5ent m0de15 f0r the
ana1y515 0f data fr0m repr0duct10n t0x1c1ty te5t5 have
5u65tant1a1 advanta9e5 a60ve the 5tandard m0de15
and mer1t 1mp1ementat10n 0n a r0ut1ne 6a515. 7he
ch01ce 0f the pr0per m0de1 depend5 0n the m0de 0f
act10n 0f the c0mp0und; a dec1510n that 15 up t0 the
u5er. 7he 065ervat10n that the n0-effect c0ncentrat10n
5eem5 t0 6e 1n5en51t1ve t0 th15 ch01ce 15 enc0ura91n9.
Ackn0w1ed9ement5--7h15 5tudy ha5 6een part1a11y 5up-
p0rted 6y 6rant PAD 90-18 fr0m the A1ternat1ve t0 An1ma1
Exper1ment5 P1atf0rm and 6y the 0ECD. 7he 0ECD made
the t0x1c1ty data ava11a61e t0 u5. We w0u1d 11ket0 thank C0r
20nneve1d, W0ut 5106, M1ke Newman, an unkn0wn referee
and the part1c1pant5 0f the 0ECD meet1n9 0f March 27 1n
5heff1e1d f0r he1pfu1 c0mment5 and d15cu5510n5 and
Annemar1e P1e1aat f0r a5515tance w1th the c0mputat10n5.
REFERENCE5
8edaux J. J. M. and K001jman 5. A. L. M. (1994) 5tat15t1ca1
ana1y515 0f 610a55ay5, 6a5ed 0n ha2ard m0de11n9. J. Em,1r.
5tat. 1, 303-314.
8urden R. L. and Fa1re5, J. D. (1985) Numer1ca1 ana1y515.
805t0n: Pr1nd1e, We6er and 5chm1dt, pp. 676.
Carr011 R. J., Ruppert D. and 5tefan5k1 L. A. (1995)
Mea5urement err0r 1n n0n11near m0de15. M0n09raph5 0n
5tat15t1c5 and App11ed Pr06a6111ty 63, L0nd0n: Chapman
and Ha11, pp. 305.
Hawker D. W. and C0nne11 D. W. (1986) 810c0ncentrat10n
 Daphn1a repr0duct10n t0x1c1ty te5t5
0f 11p0ph111c c0mp0und5 6y 50me a4uat1c 0r9an15m5.
Ec0t0x. Env1r. 5afety 11, 184-197.
H0ek5tra J. A. (1993) 5tat15t1c51n ec0t0x1c0109y. PhD°the515
Vr1je Un1ver51te1t, Am5terdam.
K001jman 5. A. L. M. (1981) Parametr1c ana1y5e5 0f
m0rta11ty rate5 1n 610a55ay5. Wat. Re5. 15, 107-119.
K001jman 5. A. L. M. (1983) 5tat15t1ca1 a5pect5 0f the
determ1nat10n 0f m0rta11ty rate5 1n 610a55ay5. Wat. Re5.
17, 749-759.
K001jman 5. A. L. M. (1993) Dynam1c Ener9y 8ud9et5 1n
8101091ca1 5y5tem5. 7he0ry and app11cat10n5 1n ec0t0x1c01-
09y. Cam6r1d9e Un1ver51ty Pre55, pp. 350.
K001jman 5. A. L. M. (1995) 7he 5t01ch10metry 0f an1ma1
ener9et1c5. J. 7he0. 8101. 177, 139-149.
K001jman 5. A. L. M. and 8edaux J. J. M. (1995) 50me
5tat15t1ca1 pr0pert1e5 0f e5t1mate5 0f n0 effect c0ncen-
trat10n5. 30, 1724-1728.
K001jman 5. A. L. M. and 8edaux J. J. M. (Ed5) (1996) 7he
ana1y515 0f a4uat1c t0x1c1ty data. Am5terdam: VU
Un1ver51ty Pre55.
1723
K001jman 5. A. L. M. and Haren R. J. F. van (1990) An1ma1
ener9y 6ud9et5 affect the k1net1c5 0f xen0610t1c5.
Chem05phere 21, 6814593.
K001jman 5. A. L. M. and Met2 J. A. J. (1983) 0 n
the dynam1c5 0f chem1ca11y 5tre55ed p0pu1at10n5;
7he deduct10n 0f p0pu1at10n c0n5e4uence5 fr0m
effect5 0n 1nd1v1dua15. Ec0t0x. Env1r. 5afety 8, 254-274.
McCu11a9h P. and Ne1der J. A. (1989) 6enera112ed
11near m0de15. M0n09raph5 0n 5tat15t1c5 and App11ed
Pr06a6111ty 37, L0nd0n: Chapman and Ha11, pp. 511.
Met2 J. A. J. and D1ekmann 0. (Ed5). (1986) 7he
dynam1c5 0f phy5101091ca11y 5tructured p0pu1at10n5.
5pr1n9er Lecture N0te5 1n 810mathemat1c5. 5pr1n9er-
Ver1a9, 8er11n.
0 E C D (1994) Draft 0 E C D te5t 9u1de11ne 202 part 11
Daphn1a ma9na repr0duct10n te5t t0 6e u5ed 1n the f1na1
r1n9 te5t.
511vey 5. D. (1975) 5tat15t1ca1 1nference. M0n09raph5 0n
app11ed pr06a6111ty and 5tat15t1c5. Chapman and Ha11,
pp. 192.