Evaluation in throughts of 2 new formulations of phoxim (25% and 40% EC, Bayer) and results of a small-scale river experiment

EVALUATION IN TROUGHS OF 2 NEW FORMULATIONS OF PHOXIM (25X AND 4O7 EG, BAYER) AND RESULTS OF A SMALL-SCALE RIVER EXPERIMENT Technical Report OCP/IRU 9O.O5 Insecticide Research Unit Onchocerciasis Control Programme Programme de Lutte Contre !'Onchocercose World Health Organisation Organisation Mondiale de la Sant6 Bouak6 - July 199O v,,or'ld Health Organization - onchocerclasis Control Programme - Insecticrcte Res€arch Unit ocP,/IRU,/90.05 Page z ABSTRACTI Authors: M. Ocran, M. Ou6draogo, C. Back Since the production of chlorphoxim, one of the operational OCP larvicides, had been discontinued by Bayer, OCP decided to test 2 new formulations of phoxim: a 40% EC and a 25% EC proposed by Bayer as possible replacement for chlorphoxim. Trough tests with the new samples were conducted at the IRU field station in Soubr6 to evaluate their efficacy. Chlorphoxim ?O% EC, temephos 20% EC and an oldeLlgBT formulation of phoxim 50% EC were tested in parallel wlth the new phoxim formulations for comparison. Results showed the 2 new phoxim formulations to have similar efficacy in troughs. The LDSO-LD95 values were Z4.S- 48.8 m9/L/s and 25.3-50.7 mg/L/s respectively for the 40% EC and ZS% EC. Chlorphoxim was more active with an LD50-LD95 of 7.3-29.0 mg/L/s and phoxim 50% (1987) was less active with an LD5O-LD9S of 29.5-66.3 mgll/s. the LDSO-L095 for temephos was 24.3-94.8 mT/L/s. phoxim 40% EC formed a more homogenous and stable emulsion when added to water than the 25% EC. For this reason, added to the fact that its solvents are less corrosive, the 40% EC phoxim formulation was selected as the candidate operational formulation. In a small river with a discharge of 1.6 m3ls the 40% EC phoxim killed 1OO% of S. damnosum s. l. larvae for a distance of 150 m downstream of application. The dosage rate was 60 mg/t/s, equivalent to 0.15 L of formulation per msls of discharge. RESUME Auteurs: M. Ocran, M. Ou6draogo, C. Back La firme Bayer, qui a arr6t6 la production de chlorphoxime, l'un des larvrcides utilis6s d oCP, a propos6 comme remplacement 2 nouvelles formulations de phoxime: 40% a.i. CE et 25% a.r. CE. Ces deux formulations ont 6t6 test6es en troughs (circuit ferm6) paralldlement avec le chlorphoxime, le t6m6phos et une formulation de phoxime CE a 50% propos6e en 1987. Les r6sultats en troughs ont d6montr6 que les deux nouvelles formulations de phoxime ont une efficacit6 6quivalente. Les valeurs de DLso-DL95 6taient de z4.s-48.9 mg/L/s et 2s.3-s0.7 m9/L/s pour la 40% CE et la 25% CE respectivennent. Le chlorphoxime 6tait plus efficace avec une DLSO-DLgS de 7.3-29.0 m9/L/s, et le phoxime SO% CE ( 1987) moins efficace avec une DLSO-D195 de 29.5-66.3 m9/L/s. La DLSO-DL95 du t6m6phos 6tait de 24.3-94.8 m9/L/s. Apres m6lange dans l'eau le phoxime 40% CE a form6 une 6mulsion plus homogdne et stable que la formulation A 2S%. pour ces raisons, et tenant compte d't fait que ses solvants sont moins corrosifs, le phoxime 40% ct a 6t6 retenu pour une 6valuation op6rationnelle. Un es.;ai en rivi0re A petite 6chelle a 6t6 effectud avec la formulation A 40% sur une riviAre d'un d6bit de 1.6 m3ls. La dose appliqu6e 6tait de 60 mg/L/s correspondant A 0.15 L de formulation par m3ls. Une mortalit6 larvaire de lOOx sur une distance de 0.15 km a 6t6 obtenue. *This document do€s not constitutequoted or translated without thc a forma l pub l ication egreernent of Wor'ld It should not b€ r€viewed, abstracted, Heslth Organrzetion. Authors a'lone areresponsible for views expreBscd in gigned art.ic.les wor'ld H€alth Organization - Onchocerciasis Control Programme - Insecticide Research Unrt ocP,/IRU,/90,05 Pag€ 3 EVALUATION ON S. DAMNOSUM S.L. LARVAE OF 2 NET{ FORMULATIONS OF PHOXIM IN COMPARISON WITH CHLORPHOXIM, TEMEPHOS AND PHOXIM sOX EC (FL6OOA) 1987 PRODUCTION 1. INTRODUCTION In 1971 and 1972 nine organophosphate rnsecticides were tested to find a replacement to DDT for the control of Simulium damnosum s.l. in West Africa (Le Berre et al. 1976). Temephos emerged as the best Simulium larvicide in terms of both efficacy and selectivity (D6joux et a|.1973, Lauzanne 1973). Therefore it was selected for control operations of the OCP in 1974. initial tests are reviewed in detail in Quill6v616 et al. (1976). Chlorphoxim and phoxim also showed some potentiaf, and when resistance to temepho-s of S. damnosum arose in 1979, chlorphoxim was selected as a replacement. Resistance to chlorphoxim also soon showed up in the southern part of C6te d'Ivoire and B.t. H14 had to be used massively to control S. damnosum s.i. populations of that area. Fu rther north resistance to temephos then occured among the savanna species of S. damnosum, but there was no cross-resistance to chlorphoxim, which remained operational until 1990 in those areas. Bayerl, the producer of both phoxim and chlorphoxim, had announced since 1986 its intention to stop the production of chlorphoxim. Replacement insecticides were actively screened by IRU/OCP, among which new phoxim formulations. Permethrin and carbosulfan had in the meantime become operational and showed no cross-resistance with temephos or chlorphoxim. In 1987 three new formulations of phoxim proposed by Bayer were tested by IRU/OCP and gave good results in troughs, but with an effective dose twice as hrgh as for chlorphoxim. (Kurtak et al. 1987). The safety margin for the non-torgr-,t fauna was very small with phoxim according to gutter test results (Wahle 196r ,. In 1988 two piroxrm formulations were tested in rivers. It was evaluated that one of them could be used operationally, but at a dose between 0.12 and 0.18 L/m3/s; and with a relatively short carry (Renaud et al. 1988). Chlorphoxim is not available anymore in 1990. and there is no readily available insecticide to replace it. Consequently, it was decided to intensify the screening of phoxim formulations by iRU/OCP and Bayer agreed to provide two new candidate formulations. The results of the evaluation in troughs and of a small-scale river test are presented below. y{orld Health organization - onchocerciasis controt Programme - fnsectrc.ide Research unlt Batch 01 2-N Batch 01 1 -N (2 litres)(2 litres) OCP,/IRU/90. O5 Pa96 { AS and 2. MATERIALS AND METHODS 2.1. Trouqh Tests Bayer provided 2 formulations of phoxim described below: 1. Phoxim 40% EC (OZ/OS/90) 2. Phoxim 25x EC (OZ/OS/9O) The 40% solvents, while petroleum. They The two new phoxim formulations were yellow color. The specific gravity of the 4Ot 0.970 at 26'C. EC formulation contains ethanol and petroleum distillatesthe 25% EC formulation contains cyclohexanon, isophoron were tested in parallel with the following formulations. 1. Chlorphoxim 20% EC Batch FL 8143 (03/90) 2. Temephos 20% EC (1989 production) 3. Phoxim 50% EC Batch FL 600A (xytol sotvent) Detailed protocol for the trough tests is provided in Annex. The tests wereperformed at IRU field station in Soub16 (Southern C6te d'Ivoire). Water fromNani6 river was used for the tests. Turbidity was between 20 - 25 JTU, pH was7'2 and water temperature between 25'c and 28'c. Larvae were collected from the vast breeding site of Koperagui on the Lower sassandra River. 2.2. Rlver Test A small scale river trial was carried out on the Nko River (s.32'N 3. 53,w)(Southern C6te d'Ivoire). Protocol for river tests ts provided in Annex, andtreatment parameters are included in Table 2, The target dose was 60 mg/L/s a.i.(= 0.1 mg/L x 1O min). 3. RESULTS & DISCUSSION 3.1. Physlcal characterlstics rdentrcal in appearance, with a pale EC was 0.950 and that of the ZS% EC When mixed with water during the preparation of the sotutions for thetrough tests, both formulations dispersed well. After 24h the 40% EC solution was stable, but a thin oily film was observed on the surface of the 25% EC solution. v{orld Health Organization - Onchoc€rciasis Control Programme - Insecticide Research Unrt OCPl I Rtr/ 9(' 05 Page 5 3.2. Trouqh Tests Results of the trough tests are shown in Table 1 and plotted in Fig 1. Detailed data and probit analysis results are contained in Figs 2 - 6. In troughs both new phoxim formulations have the same activity with estimated LDSO-LDgg values of 24.5-6a.8 mS/L/s and 25.3-67.9 mg/L/s respectively for the 40% EC and 25% EC formulations. The 1987 lot of 50% EC phoxim was less actlve with LDSO- LD99 values of 29.5-93.4 m9/t/s. Chlorphoxim was more active than phoxim with the LD50-LD99 estimated at 7.3-51.5 m9/L/s. Finally temephos was the least active of the insecticides, with the LDSO and LD99 estimated at 24.3 and 166.6 mg/L/s respectively. In operationat conditions the relatively low activity of temephos is compensated by interactions with the silt particles that provide for a very long carry downstream (Ott et al. 1989). This behaviour seems specific to temephos. 3.3. Selection of the candidate ohoxim formulatlon for rlver testlnq and eventual ooerational use In terms of specific gravity and stability after mixing, the 40% EC phoxim formulation proved slightly better than the 25% EC, but otherwise the spontaneous dispersion and activity in trough was equivalent. Considering these facts the 40% EC formulation was selected for the following reasons: - its higher concentration of active ingredient compensates for an activity which is inferior to current operational chemical insecticides. Therefore the dosage rate will be in the same range as for these other insecticides, which lowers the cost of ferry flights. - it does not contain cyclohexanone. Chlorphoxim formulations used to contain cyclohexanone, w hich, it seems, contributed to corrosion of the seals of the spraying equtpment. In addition chlorphoxim formulations had a very penetrating smell which incommodated flying personnel. 3.4. River Tests As explained above the 40% EC phoxim formulation was selected for a river trial. The dose applied was 0.1 mg/L/10, or 60 mg/L/s a.i.. equivalent to 0.15 liters of formulation per m3ls of discharge. The product was mixed in 1O L of water and applied with a Hudson Sprayer at 50 metres above the line of breeding sites. There was 1OOx mortality for 0.15 km, but there was no breeding site downstream to monitor mortality. Details of the test conditions are summarised in Table 2. Larger quantities of the formulation will be needed for a full evaluation of performance in rivers. Wor'ld Health Organization - Onchocerciasis Control Programme - InsecticidE Research Unit ocP/IRU/9C O5 Page 6 One of the main reasons why chlorphoxim was initially preferred over phoxim was the rather short carry of the latter. River tests (Renaud et al, 1988) concluded that for an experimental sOx EC phoxim formulation the operational dose would range between 0.1 and O.2 m9llx10 min. The experiment on the Nko river confirmed that 0.1 m9/Lx10 min gives 100x control of mature larvae close to the point of treatment, but the operational dose for an acceptable carry remains to be determined. 4. CONCLUSION AND RECOMMENDATIONS The trough tests in Soubr6 have shown that the new 25% and 40% EC formulations are equally active. The 40% EC formulation is preferred because of its higher content of active ingredient, because of its solvents and slightly superior physical characteristics. The trough tests confirmed earlier studies and establish that phoxim is 2 to 3 times less active than chlorphoxim. Therefore a minimum field dosage of 60 mg/L/s is to be envisaged. At this dosage phoxim 40% EC gave total control of larvae close to the treatment point in a small-scale river test. A larger quantity of phoxim 40% EC is necessary for f ull scale river tests which will determine the operational dosage and carry of this formulation. A sample of 500 L has been requested from Bayer for this purpose. Considering the equivalent activity of the 25% EC and 40% EC formulations, and the short shelf- life of chlorphoxim, a new sample of 50% EC commercial phoxim (Volaton) has been requested from Bayer. Evaluation results will be presented in a later report. World HeaIth Organizatron - Onchocercra6is Control Programme - Insecticide Research Unit Renaud, P., R. Meyer, M. Ocran, D. Kurtak (1g8g). Further formulations of phoxim (OMS 1170) as larvicides against s./. in West Africa. IRU/OCP Technical Report 4/eB. ocP/IRU/90.05 Page 7 5. LITERATURE CITED D6joux, C. & J. J. Troubat. (1973). Etude en Laboratoire de la toxicit6 sur la faune aquatique non-cible de nouveaux rnsecticides employ6s en lutte Anti- simulie. 16re et 2erie parties. convention N' v2-181-91. oMS/oRSToM du 29- xII-1 971. Kurtak, 0., P. Renaud, B. Wahle. 1987. Evaluation of phoxim (OMS 1 170) as a larvicide against Simulium damnosum s.l. in West Africa. IRU/OCP Technical Report 12/87. Lauzanne, L. (1973). Etude au Laboratoire de la toxicit6 sur la faune aquatique non-cible de nouveaux insecticrdes employ6s en lutte Anti-Simulies. Convention N' 'v2 - 181-81. OMS/ORSTOM du 29-XII-1971. Ott, D'E., R.B. March and M.S. Mulla. 1989. Adsorption dynamics of aqueous temephos emulsions onto and from r-iver bed silt particles. Bull. Soc. Vector Ecol. 14: 282-290. Quill6v616, D., H.Escaffre, B. Pendriez, S. G16baut, B. Duchateau, c. w. Lee and J. Mouchet. Control of Simulium damnosum, vector of Human Onchocerciasis in West Africa. III. Application by Aeroplane during the wet season. Methods of Application - New Formulations. IIIA. Summary Table of Tests b y Classic and Aerial Application of New Insecticides and New Formulations. WHO/VBC/76.616. evaluation of two Simulium damnosum Wah le B. 1987. Short-Term Effects of Phoxim (OMS 1 170) on Non-Target Aquatic Organisms. IRU/OCP Technical Report 16,/e7. TABL-E 1. COMPOSTTE RESULTS OF BIOA.SSAYS wrTH 2 NEW FORMUTITIONS OF PHOXIM (OMS I170) CoMPARED WtrH CHLORPHOXIM, TEMEPHOS AND PHOXIM FLoOOA (1907) - I g{, MORTAUTY FOR MATUFIE LAFIVAE f FORMULATION CST (N) cooE Correnlrgtbn lor Ueo<poure 7.5 30 TH7 i rso-os T90-r2 ! T90-t5 LC5O i uCsS (0591,c. t.)' (#%C.1.) 21.5 48.8 ' 1zt.*27 e1 (4.t47.2) 25.3(n.Fs.t) LC99 $;%tC.l.) Nbr ot T6t8@ 1n PHOXM (BAYER) (oMs r r70) &ohEc (o2oseo) LOT 012-N PHOxlM (BAYER) (oMs rr70) 25C6 EC (0205s0) LOT 01 1-N CHLORPHOXM (BAYER) (oMs 1827) 20q5 EC (le8s) LOT FL 81'tg Temp. Water pH Turbdity Conductivity lTeo-r3 | I (u5) T90-1 1 T90-16 =25o -2B. = 7.2 =2O-25JTU = 93 6 rE - 111.3 /,rS fi.7 67.6 (tto.4-77.1) | (5O &-fig5) e{.8 (5O I -l@.6 )l 51.5 ( -) m./t(-) 188.6 ( -) 3.6 (s17) 3. (85s) so.6 (s{t) o s2) 66 90.8 tw) 1 I I i I I I l ( I l_ I I I I 2 3 T9m T90-to 1.2 58S ? o) (8t 0m (3@) 36.9 (325) 3 2z2.o (@) trt.9(ffi) .2e) 71 (31 18.8 (2rr) 34.3(ul) 9,t.5(87) 7.3 2e.5(-) 21.X(-) 29.0(-)2 6G.3(-) 94.8(-) (2.1-t1.3) s5.8 (5s) 5 46 (1t N = Num0pr ol brvaeterc.d 2. Heterqeteity * tWn to albv tor a*cu&'tbn ol confi/erc* intervalc F,: 1 SUN/N/AR\' CF RESULTS CC F + i 1 o q I PHO)oM (BAYER) (oMs 1r70) 50q6 EC 1*7 LOT FL6@A rT9O-1/t T90-17 1 TEMEPHOS oMs 780 2OC,6 EC 3-76 16 t 1 I ) i I i + I TABLE 2. TECHNICAL DATA FOR RIVER TEST WITH PHOXIM 4O%EC TREATMENT DATA Test Number: Date: Formulation: Application Method: Duration of Application: Volume applied: Distance downstream of breeding sitee: Time between Application and observation: River: Discharge: Water Temporature: Water pH: Water turbidity: Water conductivity: Fpo0614A 14June 19OO Phoxim 40% EC (Bayer), batch 012-N-020L90 Hudson Sprayer, from ground 10 min 0.232 mL of formulation + 10 L of wator 5O m, 15O m 24h Nko River 1.55 mUs 250C 6.1 5 JTU 57.6 /,€ Rssults 100% mortaliU of S. damnoeum larvae at 50 m and 15O m ottr- o\E-a!90 o-vq= -"2G9)Z{- E.< ooc oo oE{OtroO_ k oH 3 8.6 * 6^^r't tr ooE d* L - e9}€\." ^ , .-.: o !?G .i 3H D oO--, o€3 re- rEv' c .; -.o,-i xoi<; lrJ -a oo o. ..6^ 6^=.3:-H x -i o tgEJo I5_jO 5 =-od't >o-OEd)9ooo_ - E6=- '!N(J o € -qr.E dEE ?, PEEE -= 'E= d Y x EasE= olL \ tJ: E oq x o 0{ o -o E o a 'a od (, o >i ,a tsn oa{ J:G io, d- 9\i ', -o q': -:F\ 6- 6rlfbo .t el t J d L d .J HH- .iR {\[:i cl l"i .1 .ri{l-II, !>;.oG-o o.tr -jio6 A:" q,L)T ra aqDd( i^ _o oI(J NI(J o(,=o. oz zllL)_U t\\o q) tot at.\\o q) bO trq ittt,*it$ii. i _.'1'i*# Ii '=e.iE-2!sl ; cY-PF sF ;Exts: 'o =-;x*IEl ii tE u;;qi €txEisE ef;ii3.; if : ii8".N Ia s€ 3r2 t ln'it5:: EE==I=rP :E;3! itrii iif$Eiri EHH iF€is s;E*!;si {iEEi +E; :; !€ Ei=:Sia *-Bi;5*1E€5: IE:saEE?i ;Hfi;:H ,Ei=' Fu;$;a!g; El=rs=E!.8;; i; qf qf f EE ?t;EiE i;iEi E!;gq!E$$ i$3EE; ; lE: g ti $is=: i'a .ssixII niiEsi iggistriE sliiBgi&5€ N cc : C o oI d cl c c. @ o -o oBO\t. 3<o /- -; 'O -- >.rlE zi o0-o -Ei.- ^9'o 66v* .t -r o :6lr=s: =v!, o@ .I "3€3 o -o o o C' 6o t) O U) l& 6 o c 2 E -}( (g E o 6 >\@ -o@ rr o (! o L oL 6 60\ .9)St a(\ =o- oh!u -cxi xo.r E sQu- -i >::EU! ot- d o- ul ci eat- ,q \o *.ul c3 .E cir- a2 o GI o E E xo a o, q o E E o O o E.oo q o! =, qt o 6t o oz Q Lo 1Jo() a! L c t\ E 6 E E 6 a.t o dE ot ei€ RP a6o-'7 of 1xr! n<-.rQZ (t)r- o\ x o n o(n r! t LJ AC>9 vo o(, tr^ X>rotr =oo-Eo- N. ?D (P j & -t rD r* SD 5 d J J -I .J u, c-l a, tY J + 5 * _o ! It I I I Fi I I I I t I ti{bift\o t\ q) 60 s\ fn cri ! v J2 oo -o 't, o c E^F€ .qRo-9\Og\o E. €6 =' 8s=-oo.lJ-P .Etx cs .oJ}ED 6Ql !E99 gp -\ oqJ .i t) .i *s oo e io .i({o -tr:l -ai ^ aos '-ca 2 5luUG=cz .9 -dEE() 5J N.A5co .9io - 9o.-J aq-tr o\o Cr O\h- /\ (JgJ Eo x€ 'i ood<J oo--:E\9-oo X-|*E-h S tsq;ggq otE )Yo-o - oo5EE-o!-6 0* '. i 6oOc)r JE \:/F;dEO !gE: n*$N'E tr ,t9o :? oo-i oE r .€: .E.E:&E -6. €EO o-:g.?; ?5-\L .Y o OL<Ee € L ol& h .: 5 =6(/) ;.U o o al 5OAC?o qtr:, a) -o G Va E +TE>o tr 00 q .o(J .o6>. E =o c, 556 q .2x. I!.o .: EE7 ..\O O9dE q\t O\oL' =\oo ! :6 q) :!' oLoO . -l( rct\ o =od 2otrqt o oc\ rc6. OE I 6()y AO e60 ': <s t o lC o o o s,(,) o o :, oq ri. a o xo q o () o tJ) t! (r) ca Iu ruo 0.oz il vl z(J Ea.ro € c{ o Fd8d(a mO- E3 odE z3 Q=d EUQZ o 'ti a{ 5 t o L q o c{ t\ v-c o9tr6r;Ytr 6t, tro E EoO q) =n lu E oz xo a o U ho o o, .9 x oF o f o q 6.o 'iltd,,rt.i,&. ifi lt (-l a.lo o\ € al o- sl o o dd 1.| E 6Lo o c.t \ Frq. 2 RESULTS OF PROEtT ANALYSIS 5 iterations CHI-z CRITERION = 0.1 HETEROGENOUS DATAIog LD99 = L8l2L8 l.?06 ( < 2.003LD99 = 64.89062 50,802 ( < 100.593 Iog LD95 =LD95 = I 48 Iog LD50 = l. 3E552LD50 = 24.{638{ ( 1.828 < 67.283 1.329 < < 1.44421.338 < < 27.787 s.E.b 0.167 DATA TRANSFOR}'S 1.607 (40.412 < 6b80? 76107 ,.ntercept s Iope a b -? .625 5.491 DEGF =CHI-2 =Probabrlity = HETER =T-VALUE= GFAC = 8 153. 365 0. 000 19,171 2. 310 0. 094 TITLE:C9006044 PHOXIM 401 EC LOT 012-N (02059U) (BAYER) LODE tRLr Cgi)i2SUBTITLE:KOPERACUI, R. SASSANDRA 04 - 07.06.90Dose unlts:t{C/LControl : 0.0 : DOSE TESTED DEAD LOG CORR DOSE DEAD CORR. PRED. CHI-SQ PROBIT PROBIT CONTRIi coft i PEHtDATA * L,778 t,778 1.77a L.4i7 t.47 7 1.47 7 L,477l. 1?6 l.176l. 176 0 0 0I 5 1 8I 7 2 oBs 99. 00. 00. 66. 78. 77, 46. 2L.' 15. 4." I 2 3 4 5 6 7 8 9 l0 60 39260 38s60 16430 34230 28330 24030 3?715 301t5 31315 333 368 385 r64 226 223 155 153 66{9 l4 43 l5 99 42 l9 259l 73 388. 384. 163. 7,319 6.012 7, r39 ?. t39 ?.139 5.486 5.486 5..186 5. {85 3.834 J, 334 1.834 i,7 5. ,l 5.7 5.7 4.9 4.2 3.9 3.2 0 5 0 0 0 0 0 0 0 226 223 185 153 66 49 () ! 1 I IJi 99, u 99, s 99. 7 6b. I 7!, \ ??.1 It). 92t.9lr. i 88 36 't6 06 50 90 76 l4 26 .80 3. 55 I9.77 c9€rxg4A H.OXIi 4O7 EC LoT 012-H (@9590) (8d/ER) CDE rRU C9012 Kopmqcul, R. sASsAloRA 04 - 07.06.90I 19 -i_: 7 -lc 19C 6 1 .{ F c d o, {. zo o. L cr JL --l 3 - 1l- 2 -2 B a Loc DosE ( rn r'lc/L ) Fig. 3 NESULTS OF PROBIT ANALYSIS 6 rterat rons CHI -2 CRITERION = 0 ' I HETEROCE!{OLIS DATAIog LD99 - I.83017 1.705 < < 2,O77 LD99 = 67.634114 50.?50 ( ( ll9.5l7 Iog rP95 =LD95 = Iog LD50 = 1,40237LD50 . 25.25603 1.70484 1.607 ( ( I.53750.68095 40.433 ( < 77.Ii8 L316 ( ( 1.,178 20.800 ( ( 30,086 int ercept slope a : -2,626b . 5.{38 S.E.b . 0.15{ DEGF =t:Ht-2 -Probabrlrty = HETER =I -VALUE- GFAC. 8 249. 320 0.000 31. 165 2.310 0. 133 TrTLE:C9006048 PHOXIM 25: ric LOT 0rl-N 1020590) (BAyER) CODE IRU C90lt SUBTI TLE: KOPERACUI , R. SASSANDRA 04 - 0? . 06 , 90Dose unrts:MG/LControl : 0,0 Z DATA TRAN5SOR5S OBS LOG DOSE CORR DEAD CORR. PRED. CHI-SQ. PROBIT PROBIT CONTRIB. CORB. PERC.DATA DOSE TESTED DEAD t.778 C9S604B PtDXIl{ D4 EC LoT 011-t1 (028590) (SflYER) CSE IRU C?€lt KopERACUI, R. SASSATORA 04 - 07.06.90 o -2 60 30 30 30 15 l5 15 7,5 544 42L 335 3162ll 404 r50 347 355 345 s42 42L 334 239 158 L52{3 37 32 4 78 7A 77 77 76 ?6 7b ?5 542.0 420 ,5 334.0 239.0 158. 0 ?.680 8.039 7,749 5.695 5.671 ,1 .685 1,437 3. 755 3.659 2.730 7,O43 ?.043 7.043 5,407 5,407 5.40? 3.770 3.?70 3.770 2. 133 7.66 7 .82 5,L2 13.62 r. rb 142.31 .r8.5r 0.02 1.37 r5.16 t I 3 4 5 6 7 a I 10 60 60 152.0 43.0 3?.0 32.0 4,0 1., r.7 r.4 l.{ r,4r.l r.1 r.1 0.8 99 100 99i, 71 37. 2A. 10. 9,l. 6 0 6I (; 7 7 0 2 6 9 7 6I 6 7 7 0 2 99 99 99 75 74 37 ?8 IOII I ? 6FI .To ; aZo o(4U c! o< 7e 36 1e I 2 E 3 a B ) 4 tOC t€SE I :n,{C/L I Fig, I Iog LD99 =LD99 = 5 r.71 1,47 log LD95 = 1,46305LD95 = ?9,04326 RESULTS OF PROBIT ^NALYSIS5 rterat ions CHI-2 CRITEITION = U. I HETEROGENOI.'S D.1 TA1.324 ( ( 4.53321.090 . < J{123.05:' 1.166 ( 3.42714.647 < ( 2671.{,85 S,E.b 0. 130 :PHOXIME 2O'" EC LOT !'L6I+3 SASSAT\iDRA 0t - 0i /06 190DATA TRA\SFORMS 556 160 Iog LD50 LD50 = II 63r367 92 ( < l.l55( ( l{.301i.2 0.38s2 ,126 lntercept a : s l,)pe b = DEGF =|HL-Z =Probabr.lrty. HETER. T - VALUE= GFAC = 1 8 0 4 0 2.633 2.742 29 .04 0.00 14.52 4,30 0.60 T I TLE: C9OO604D SUBTITLE:KOPERAGU Dose unrts:MC/LControl i 2.0 X CHLORI, R. (03/90) IBAYER) CODE IBU DATA DOSE TESTED c9m6e4o cHLmpHoxtnE 202 EC toT FL8t43 (03,196 I r3rr,iE[,r CoDE IRU CSli(OpERAcuI, R. SASSA|DRfi 84 - E7,€6r,:0 l0 36615 3r9i,5 3.\l3.75 399 362 229 2t2 9rl 1i ?6 75 i1 36r,I 7227.2 s208.6 587.9 I 560 rtz 229 6.683 5.858 s.033 t,207 10. l3 17.30 1.53 0. 09 7t,2 54.5 22.O DEAD OBS 95. 7L 55 23 LOC DOSE CORR DEAD CCRR. PRED. CHI-SQ PROBIT PROBIT CONTRIB CORR, PERC. s t 2 3 4 I 1.45 1.1{ 0.66 0.5 98. I283 o 1 o F @o(I o Z r c, x.6I ,-A u ct 6 o 70 5 4 I 1,' I 'iro ,l : J I 4 z -c 0 LOG reg ( rn ttC/L ) Frp.. 5 RESULTS OF PROBIT ^NALYSIS5 iterations CHI-2 CRITERION = 0.1HETEROCENOUS DATAIog LD99 . 2.22L60 ERR LD99 = 166.5?02 ERR log LD95 = I.97689 ERRLD95 = 94,81763 ERR Iog LD50 = 1.38626 ERRLD50 = 24.3366? ERR lntercept s lope a. b I 2 139 785 S.E,b = 0.193 DEGF =CHI-2 =Probabrlity =HETER = T-VALUE= CFAC = 1 61.000 0.000 6 1. 000 L2 .7 LO 47.309 TITLE:T900605c TEMEPHOS ( I989) 202 (KORHOGO)SUBTITLE:KOPERAGUI, B. SASSANDRA 05.06.90 (PROCIDA) CODE IRU C9OI4Dose units: t'IG/LControl i 2,1 I DAT,{ TRANSFORMS x OBS LOG DOSE CORR DEAD CORR. PRED. CHI-SQ PROBIT PROBIT CONTRiB CORR PERCDATA * I 2 3 60 29730 41015 324 281 194 L22 94. 6 47 ,3 37 .7 78 /b z8 I8 I1 o.7 9.1i.i 6II 599 904 650 6.091 17.115.2s3 32 . 50 4. { ls 1l . 39 94 46 36 DOSE TESTED DEAD Tgffiorc TtltEpHG (1S9) KPERMUI L,7 1.4 1.1 5 2 3 20tr (K0$nG0) (FR0CIDA) CIDE IRU C9014 , R. SA€StrtoRA 6.S.90 9' t,, 1e€ I I i 30 6 .t F oo & rq) E I) d +10 .l 5 I 1, I f I -z 3 2 c 0 c LOC Df'E ( rn l{CiL ) o Frg. 6 RESULTS OP PROBIT ^NAI,YSIS6 r.teratrons CHI-2 CRITERION = 0.1 HETEROGENOUS DATAIog LD99 = 1.97030 ERR LD99 = 93.38928 ERR log LD95 = 1 ,8237 1LD95 . 66.63646 Iog LD50 = 1.46991LD50 = 29.50527 ERR ERR ERR ERR intercept s Iope -l,,,344.649 s.E.b.a=b= DEGF =CHI-2 =Probability = HETER = T-VALUE: GFAC = I 531 000 531 ?10 693 Dose unrts:Mc/LControl i 2,0 Z 0 .229 DATA TRANSFORMS LOC DOSE CORR. DEAD LOG DEE ( rn ttCi L ) 55 0 55 L2 2L TITLE:T900607C PHOXIn (193?) 501 cE (369226) LOT EL 600A (596A) (BAyER)SUBTITLE: KOPEB]IGUI , R. SASSANDRA 0?,06,90 oBs CORR. PRED. CHT.SQ PROBIT PROBIT CONTRIB CORR PERCDATAI I z 3 -2 DOSE TESTED DEAD T9006O7C ptlilItt (1*7) 5BZ CE (e69%) r0T FL 690A (596,A) (BAVER) KmER*Ut, R. SASStrt0RA 07.06.98 a/ 60 53830 32515 228 s16 L24 42 95.9 138,2 118.4 I 776 477 176 15.6 20.0 38. 3 6. 732 6. {33 3.964.666 5 .034 27 .O3 4. 038 3. 634 19. 64 9s.8 36. I 16.8 F Itro Z b o Z,oI n !I 6 I 2 2 i 'lss 1ge i i lzo I 5 30 I 1.1 I i I 3 .,1 0 I l' I + I B ANNEX OCP PROTOCOL FOR CLOSED-CIRCUIT TROUGH TESTS BRIEF DESCRIPTION. Thia mcthod, Hhich Hr8 dcvclopcct for OCP by Dr. H. Jamnback, is baEect on e syatem that h€ and Mr. R. Gaugler uscd .xt€nBivcly in Nck York gtrt.. It ha6 b€en further modified by the OCP Insecticicte Re- B€arch Teen in Loio, Togo. Larvea col'lectcd in thc fie'ld rre allowed to attach on a r€movablc plate 'lying in thc bottoot of a trough where Hat€r is r€circulatad by a pump. The plate is transfgrred to a s.cond trough where thc ina€cticidB has e'lready been mixed into the recircu'lating wrter. After 10 ilinutes the platc ie trensferrcc, to a third trough with clean Hat€r. l.{orta'lity is countect after 24 hours. TECHNICAL DETAILS. The troughs uBed ar€ constructcd of aluminiulr and painted xhlte. The troughs are Eo cm 'long, 15 cli wide and I cm d€ep. They are c]osgd at the uppar end. At the low€r end, a conEtriction or'lip, 15 cm long, 8.8 cm wlde end 2 cm deep is attrched. A removable metal p'latE reats in the bottofi ofthi6 lip' water ia pumpcd into th€ clos€d end of the trough and flowa over th€ plat6 in the lip into a reEervoir, to bo aspirated by thc pump again. SmaIl e]ectric centrlfugal pumps with a crischerge of ebout 22 L/mln are uaed, end the velocity of water ov€r the rttachilent plat€ la about 70 ci./a. For the attachnent and observetion systems, the reEervoir is a plactic waEte-bucket holding 60 L of water. For the exposure System, a 10 L enam€lware pot is ug€ct at a reasrvoir to raduce th€ amount ofinsecticidc nsedod, Rivor wet.r ia usod rt rIl atrgor for routina tarts. Its chtrlcterirtics(temperature, pH, turbidity and condqctivity) arc measurcd and noted. Larva€ ara collacted by removing their naturr'l aupporta frdr thc riv€r. Thay are transportcctin p'lastic bags p'laced in rn lcebox. cero is taken to 6€paratc th€ larvae from the ice by a layer offresh leavos or graaa, Although thc equipm€nt can bc us€d at thc riveraidB using a portable genera- tor, this type of tclt iB usua'lly don€ in a 'laboratory which may ba at Bome distanca from thc col-lecting point. The vegetrtion rith the rttached larvac is placccl in the attrchment trough. The larvae natu- rel'ly ]eave the vcgetation to Beek a Btrong€r currcnt anct thus attach to thc p'late of the 1ip, xhich has the highest valocity ln tha trough. The lrrvae ar6 left for 24 h to enEure firm attachment. Lar- vae ihich pupate during this period ere eliminated just before exposura. The plate uaue]Iy containg about 10O oatur€ 'larvae and sev€rr'l hundred young larvae, The larvicide formulations are approprirtely diluted with clistilled water immedJltely prror to the test. The quantity neceasary for the lowcst concentration is added by pipette to the r€s€rvoir of the exposure trough r{ith tha pump running. The ina€cticide is alloHed to run for 5 min to obtain a uniform diapersa'|. Additiona'l lnsectlcido iB added to th€ Bame syEteil to obtain th. highar concentrr- tiona in a tests Geri€t. The plat€ from tlre ettachment trough ig now carafully lift.cf out and plac€d in the cxposure trough for a defirrd pcriod, u3ually tO rin. Larvae cletaching Gturlng tho oxpoaur. pcriod rre cought in e ned rttach.d to thc lip of the trough. After thc cxPo3ura pcrlod thc pl.tE and tho n€t ara tranaferr€d to a holding trough. At the end of e 2a h observation pariod, th€ detach€d lervae in ths net, as well as eny dead onea still at- tached to the plate ar€ prBserved in alcotrol for counting. A first collection is generally made 2 h eft€r exposure Eince most ]ervre c,etach in this p€riod anct the'larvae ilty decompose if left ct€ad too 'long' Th€ living larva€ ere klllect witlr hot Hat€r and also preEerved in a'lcohot for counting. The number of expoaed, living, and d€rd lariae {6 estrb'lishecl separat€ly for mature 16th-7th inBtars) andyoung (lrtt-sth) larvas. Larvea exposad to incocticide rnd pupatlng cturing thc teste rrc not countod cxc€pt for thc control ih.rG they ara includsct ln th. nuf,ber Burvivlng, The usual conlormation of a te6t6 i6 r range of 5 conc.ntrutions and en untraeted control with 2 rcp]icate3 tt arch conc.ntrltion. P.rcent mortrlity iE cr'lcu'latcct et erch cono.ntretion lnct th€ concentration/mortallty relrtion is oBtrbllsh.d by probit analysia. ANNEX OCP PROTOCOL FOR RIVER EVALUATION OF INSECTICIDES BRIEF DESCRIPTION An adequate etr€tch of riv€r is firat aurvey€d on foot, by boat or by he.ticopter. potentia.l breeding sites are proapected for the presancc of eggs, larvae and pupae of Sinuliun dannosun s,1. Individual supports (IEaves, blades of graBs, branches, roots) are marksd xith numberect co.loured plastic strips. The d€naity of ]arvag ia noted for each support. Relevant factors including t€mp€raturo, turbic,ity and pH of the wBter are recorclBct. The breeding sitEs arg nappcd and their distance downstrean from treatment point calculated. wh€n the prospection is done from ground lev€1, a threed diBtance metor is used to ce'lculate distances downstream of brecding sit€s from the treatn€nt point. The discharge of tho river at or nelr th6 point of treatment is dctemined either by reading a water gauge (if one {g available), or by measurements Hith a curr€nt [€ter. The amount of formulation rEquired to give the desired dos€ is then ca'lculated. It is usually dilutBd with a convenient amount of river water just befor€ application. Th€ actual epplication may be cerried out by pouring or with r hend prosaur€ sprayer in the case of Bmall atreamE. A boat and a pierced drum may also b€ used, In th€Be tHo caBeB, th€ app'lication can b€ timed to'last 1o minutes. The application may also b€ done from the air using r helicopt€r. Th6 inBecticid€ is then sprayed fron the cabin with an ext€nded nozz'le, or Bprayed into the river using the operational spraying system rhen the vo'lume of insecticide axceeds 15 L. The insecticide is applied in one or severa'l swaths covering from bank to bank at some distance above the first breeding site in such a way that the insecticide wavB covera uniformly th€ width of the river. The cffect of thc lns€cticid€ treatil€nt iE monitored 21 or 48,h after application. Larval density ls .vrluetcd eg.ln for cech of th€ previously 6.rkGd Bupports, when posEibIe, aamples in carnoy ar6 taken for cytotaxonomical id€ntification, susceptibi Iity tests are carried out with current OCP opgretiona'l insecticide6. and