Cytotaxonomy of S.damnosum complex: January- December 2001

|t- IIt WOR.LO HEA.LTH (,R,GANIZA.TI(,N A'IGNI@6N (,RGANISATI()N MONOIALE DE LA. !iANTE IE'Oi OE L'ftQUC t @ SUBJECT OMS / ONCHO, B.P. 36 KARA, REPUBLIQUE TOGOLAISE T6t. 60.60.75160.06.26 (Fax.60.05.26) Cytotaxonomy of S. damnosum complex - George K. Fiasorgbor (Technical Officer) - Coulibaly Yaya(Technician) - Sedou Naniogu6 (Technician) PERSOI\NEL: SIGNATTIRE OF RESPONSIBLE OFFICER : PERIOD COVERED : January - December 2001 DATE OF SUBMISSION : 08 February 2002 CONTENTS 1. INTRODUCTION 2. CYTOLOGICAL IDENTIFICATIONS 3. MATERIALSA{ETHODS 4. RESULTS 5. TT{E DISTRIBIJIION OF THE CYTOSPECIES OF TFIE SIMULIUM DAMNOSUM COMPLEX IN THE YEAR 2001. 6. RESEARCH CONTRIBUTION FOR THE JPC 2OO1 : ZONE EAST 7. OTHER TECHNICAL REPORTS 8. MEETINGS 9. OBSERVATIONS/COMMENTS lO.ACKNOWLEDGMENTS 2I. INTRODUCTION The present report covers the period from January to December 2001. Throughout the y"ur, -u.h attention was concentrated on the laboratory rearing of the different members of ih. S. du.r,orr- (s.1.) complex started in March 2000. The study finally ended up in July ZOO1, tt u. "o*"ing a total of l7 months. Sufficient larvat and adult samples of S. damnosum (s.1.) complex were realised during the four main seasons of the year : 1. Rising water level (Mont6e des eaux) 2. High water level (Hautes eaux) 3. Low water level (D6crue "periode d'harmattan) 4. Lowest water level (Etiage) Cytotaxonomic determination of the reared larvae and morphological identification of the adult flies were carried out by members of the Research Team at Kara. The greater part of the reared samples (larvae and adult flies) were however sent to the DNA laboratory in Ouagadougou for-DNA determinations and also to be kept as reference samples for the future. 2. CYTOLOGICAL IDENTIFICATIONS Apart from the cytological determination of the reared larvae as stated above, routine identifications were also carried out on the lanrae of S. damnosum (s.1.) from all the river basins still under larviciding control in the entire onchocerciasis Control Programme Area' In the East, these include the five major river basins (Volta, Mono, Asukawkaw' Oueme' Niger) and their tributaries. In the west, the rivers include the Niger in Mali, the Niger in Guinea (Conakry), the Milo in Guinea (Conakry) and the Sassandra in lvory Coast' Some identification results of reared larvae from catching sites chosen in the initial Programme Area were as well embedded in this report' In the year 2001, a total of Two Thousand one Hundred and Eighty Seven (2187) larvae of S. damnosum 1s.i.; were cytologically identified by the two cytotaxonomy teams of the OCP (Kara and Odienne). 3. MATERIALS/TIETHODS Materials Specimens on which this report is based came from three (3) main sources : a) Specimens from routine checks by the operational Bases. b) Specimens from susceptibility tests. ;i i'p""i-.t t originatingfrom the laboratory rearing of members of the S' damnosum (s.1.) comPlex Methods Larvae of the S. damnosum species complex were preserved in carnoy's fixative (3 parts absolure alcohol f p*t glu"i"l acetic acii;. The samples were stained by feulgen and chromosom. pr"p*utions made as described by Dunbar (1972\ and Boakye (1988)' Identifications were made using the chromosome maps of Vajime and Dunbar (1975) ; Meredith et al. (1986) and Surtees et al' (1988)' I 34. R.E,STJLTS CYTOLOGICAL IDENTIFICATIONS (200 I ) 5. THE DISTRIBUTION AI\D MOVEMENTS OF THE CYTOSPECIES IN THE YEAR 2OO1 S. sirbanum : As shown on the distribution map and the summary of the species composition E-y "ytotu**omic identification, S. sirbanum was recorded on all the river. basins still under larviciding control in tte y"u.200L Lik" in the past this species has the largest distribution range in the northern part of the Programme. It makes dynamic movements towards the south in the dry season, especially in the harmattan' S. damnosum (s.s.) : In the year 2001, S. damnosum (s.s.) was registered in sympatry with S' sirbanum on atmost all the river, *ith th. .*""ption of the Pru and its tributaries on the western flank of the volta Lake in Ghana and also on the eastern tributaries of the Niger River in Benin (the Sota, the Alibori and the Mdkrou)' As usual, S' damnosum (s's') was found mainly in the transition zone between the forest and the savanna' S. squamosum : Was observed in the year 2001 on the eastern tributaries of the Volta Lake in Ghana and Togo and on the Mono and its tibutaries in Togo as well as on the Sassandra in C6te d'Ivoire. S. yahense : As in the year 2000 the breeding site of Tsatsadu Falls on the Tsatsadu River in C[i-u i. tn" only poinifrom where S. ]rahense was again registered in 2001' S. sanctipauli (s.s.) : was recorded at Bac-Semien on River Sassandra in c6te d'lvoire' S. soubrense : The "Beffa" form of S. soubrense was observed in the year 2001 on the Oueme River in Benin at Agolin-Pahou ZONE WESTZONE EAST TOTAL TDENTTFIEDBASINSTOTAL IDENTIFIEDBASINS r52 260 63 Niger (Guinea-C) Sassandra Milo lger 131 391 145 29'.1 Asukawkaw Volta Mono Oueme 5s8r629TOTAL 2187GRAND TOTAL a 46 RESEARCH ACTIVITIES IN THE EASTERN ZONE JPC 2OO1 The Applied Research and Environmental Monitoring Team (AREM) based in Kara, Togo carries but research in four main disciplines in the year 2000. These are (1) Larval Cyltaxonomy (2) Laboratory Rearing of members of the Simulium damnosum complex (3) Insecticide Tests and (a) Fietd Training of National Entomologists in OCP Vector Control Unit Entomological Evaluation methods. 1. CYTOTAXONOMY As compared to other methods of identification (morphotaxonomy, DNA probes, Heteroduplex Analysis etc), larrral cytotaxonomy still remains the most reliable means of identiffing members of the Simulium damnosum (s.1.) complex. As a result, OCP depends heavily on "ytotoonomic identification results for both operational and research purposes. The inversions recorded on the banding pattern of the giant salivery gland polyene chromosomes could be utilised to determine resistant cytospecies and also to follow up the movement of reinvading flies (Study of Insecticide Resistance and reinvasion). CYTOTAXONOMIC IDENTIFIATION RESULTS (2OOO I In rhe year 2000, a total of Eight Thousand Three Hundred and Ten (8310) larvae of S. darnnosum (s.1.) were cytologically identified by the two cytotaxonomy teams of the OCP (Kara and Odienn6). RESULTS CYTOLOGICAL IDENTIFICATTONS (2000) ZONE EAST ZONE WEST BASSINS TOTAL IDENTIFIED BASSINS TOTAL IDENTIFIED VOLTA 2767 MILO 104 MONO r532 SANI(ARANI 208 ASUKAWI(AW 338 NIADAN 74 OUEME 2096 TINKISSO 318 MGER (B6nin) 154 MGER (Mali) 131 KABA 6 SASSANDRA 219 NIGER (Guinea) 363 TOTAL 6887 1423 GRAND TOTAL 8310 5YEAR 2OOO S. sirbanum : In the course of the year 2000, S. sirbanum was registered on all the river b*i". in th. OCP with the exception of the Kaba river in Guinea and the tributaries of the Volta Lake in Ghana/Togo. It has a large distribution range in the northem half of the Programme and makes periodic movements to the south in the dry season, especially in the harmattan. S. damnosum (s.s) was recorded in sympatry with S. sirbanum on almost all the rivers in the OCP "*-pt o, the Kaba in Guinea, Niger in Guinea and the Niger in Mali. It is found mainly in the transition zone between the forest and the savanna. S. squamosum: This species was observed in the West on the Kaba, Niandan, Milo, S-k**i *d Sassandra. In the East, it was registered in its natural habitat in the mountaneous forest regions east of the Volta Lake in Ghana and Togo. The main rivers are Asukawkaw and its tributaries, the Mono and its tributaries, and on the tributaries of the Kara and the Mo. Few cases of S. squamosum were also found on fuver Zott, a tributary of the Oueme River in Benin. S. vahense : The breeding site of Tsatsadu Fulls on the Tsatsadu River in Ghana is the only point from where this species was identified in the course of the year 2000. S. soubrense : The "Beffa" form of S. soubrense was observed in the year 2000 on R. Mono at f*ar-pto* in Togo. In Benin it was registered on the Oueme fuver at Agolin-Pahou, M'betekoukou, Atchakpa, and Gba. On the Okpara, it was found at Bassa-Barrage and Kaboua. No case of S. soubrense has ever been recorded in Ghana since the inception of OCP operations in the country in 1974. S. sanctipauli : The "Djodji" form of S. sanctipauli still remains extinct in the Eastern Zone .ince tg88. Before this date, this species was recorded mainly on the Asukawkaw River and its tributaries in Ghana and Togo. 6CoMPoSITION DES ESPECES (2000) CYTOTYPESBASSINS / RIVIERES S. squamosum S. sirbanum S. sirbanum, S. damnosum (s.s.) S. sirbanum, S. damnosum (s.s.), S. squamaosum S. sirbanum, S. damnosum (s.s.), S. squamaosum S. sirbanum, S. damnosum (s.s.), S. squamaosum S. sirbanum, S. sirbanum, S. damnosum (s.s.) S. damnosum (s.s.), S. squamosum, S. sirbanum S. sirbanum, S. damnosum (s.s.) S. sirbanum, S. damnosum (s.s.) S. sirbanum, S. danmosum (s.s.) S. sirbanum, S. damnosum (s.s.) S. sirbanum, S. damnosum (s.s.) S. damnosum (s.s.), S. sirbanum, S. squamosum S. sirbanum, S. damnosum (s.s.), S. squamosum S. squamosum, S. yahense, S. damnosum (s.s.) S. squamosum, S. damnosum (s.s.), S. sirbanum, S. soubrense S. damnosum (s.s.), S. squamosum, S. sirbanunt S. damnosum (s.s.), S, sirbanum S. damnosum (s.s.), S. sirbanum, S. soubrense, S. squamosum S. damnosum (s.s.), S. sirbanum, S. soubrense S. damnosum (s.s.), S. sirbanum KABA MGER (GUINEE) TINKISSO NIAI{DA}I MILO SAI{KARANI MGER (MALr) BANI SASSA}IDRA PRU DAI(A WHITE VOLTA OTI PRINCIPAL KERAN KARA MO LAC VOLTA HT. MONO BAS MONO HT. OUEME BAS OUEME OKPARA MGER EST (BENIN) 7[T =UJ :D! o a -t oz o =+, I n7< =\tr, -_\ : I I I I I Fa UI FJ o o j lJ.lr oz o E a m ,\ t I I=(L:5 r o o aJ ,(t oooN o UIJJ uIf z z o lu o I =o z o x F ol- o o z o IF o I lJ. F z lII a I ,,1 ? .(-\rj ''rr i- '(I \l l\- <1 I I I / \\,\ :\ x I( ! Itl ,l - t' ,., \ I \ I r\, ) ,/).. / -,i r,, .r() -ir EI. o_ 4\ /" /\ ( I( .4 s( ejt' - ',,,' 'l^ { -tIs i1 a (-l 'r t \(/ dt oo I ( t \ !- \ ri\(\ tJ- z<..)( i I I I)\. '__ / _ ,l/iI[_ -\ ,'t i ,: !.:- l;.! ( .xt '*,=, \ E: IIJ..o) \ I ,.\ 'v' A,/.arr (f {\ 3-.5H<EO<AU)o> EIII ,' Ir .oi' -)\r _\-) l) ,r-E- oP :d 3e 'ee a-b o€ &:5rog \cos \_. 82. LABORT{TORY REARING OF MEMBERS OF THE SIMULIUM DAMNOSUM COMPLEX IN THE YEAR 2OOO. Single egg-batches of three members of the S. damnosum (s.l) complex were successfully ."rr.d to adult flies in the laboratory in 1995 and 1996. These were the Beffa form of S. soubrense from M'b6t6koukou and Kaboua on the Ou6m6 and Okpara rivers in Benin; S-. yahense from Tstsadu Falls in Ghana and S. squamosum from Kouniohou on R. Ban-Hou in Togo. The main objective of the rearing exercise in 1995 and 1996 at M'b6t6koukou and Okpara was to p.ou-id. enough samples pf dark-wing tuft adult females and larvae of the S. damnosum (theobald) comple* forDNA, taxonomic and cytotaxonomic determinations. Before this date, these dark-wing tuft adult females from the Ou6m6 and Okpara rivers in Benin were scored by the DNA technique iui savanna cytospecies (.S. damnosum (s.s.) and S. sirbanum). The DNA determination was contrary to all previous cytotaxonomic ia*tifrc"tion of the same populations as the 'Beffa" form of S. soubrense, which infiltrates into the OCP area from Nigeria each year during the rainy season. It was therefore an attempt to investigate this discrepancy between the DNA and cytotaxonomic identifications that VCU hierarchy called for the rearing of wild-caught females of the S. damnosum complex in the Ou6m6 Basin to provide more samples to confirm once and for all the actual identity of these biting adult females. Rearing work was also undertaken on the Comoe river in Zone West between 1995 and 1996 with S. sanctipauli (s.s) as the target cytospecies. In the year 2000, the same exercise was carried out in the two zones (East and West) with the saml objective. In addition, the rearing was supposed to cover all the various members of the S. damnosum complex in the OCP area. This was to provide enough reference samples as the Programme draws to an end in2002. We report below the full cytotaxonomic identification results of the study in the different river basins in Zone East. 9PROGRAMME D'ELEVAGE DES SIMULIES EN ZONE EST SAISON SECHE DECRUE (Periode d'ha rmattan) ETIAGE ASUBENDE R. PRU SUGU R. VOLTA BLANCHE EKUNDIPE / Doqonkade R. DAKA TSATSADU.FULLS R.TSATSADU BAGAN R. MO TINKIRO R. ASUKAWKAW LAN ZANDA R. KARA KPOBA R. MONO TITIRA R. KERAN LANDA MONO R. MONO TOUTIONGA R. OTI MBETEKOUKOU R. OUEME BATIA R. PENDARI KABOUA R. OKPARA KEROU / KEREMOU R. MEKROU BETEROU R. OUEME GBASSA R. SOTA AL SAISON DES PLUIES MONTEE OES EAUX HAUTES EAUX ASUBENDE R. PRU SUGU R. VOLTA BLANCHE EKUNDIPE / Doqonkade R. DAKA TSATSADU-FULLS R.TSATSADU BAGAN R. MO TINKIRO R. ASUKAWKAW LANDA-POZANDA R. IGRA KPOBA R. MONO TITIRA R. KERAN LANDA MONO R. MONO TOUTIONGA R. OTI MBETEKOUKOU R. OUEME BATIA R. PENDARI KABOUA R. OKPARA KEROU / KEREMOU R. MEKROU BETEROU R. OUEME TOUTIONGA R. OTI TITIRA R. KERAN r'--' t. I l0s 6 o o o o o o o o o o o oo o o o o o o o o o utI o o o o o o o o o Doo o o o o o o o o $ 6u o o o o o o o z =o o o o o o o o o o FoF o o o o o t = UJ UJ Llt TIJ uJ ul TU uJ IrJ ut E ozo t- r() ci t-lo ci t- ro C' Fro ci t- r^() C' t(,I\ C; t{)\o tot\ o rot- CJ o F tr to(Y) C; to(o o lo(r) Ci ro(r) c; lo(7) Ci @od @o c(i @q @ @o C; =UI ozotrf,oF (\l ozo trf,o (Y) oz o l o \r ozo Fl oF lo oz o Ff, o o E, Yz F (r) o E, !zz F t oE Yz F r() o G, =z IJJtrt! = tr tro tro tro tro tro 3g 3 $la 3g 3g la =g =g fa =g 3g :)o l.l.g dt oo(o o oo(oFo oo(oFo oo(oFo oo(oFo (\t (Y) ol(Y) ol(Y) 6lc) J =oo to FJo Jo Jo Jo FJ o =g tsg lo =$ =sfo =g =g :)o =g =sf,o uJ o oo c.jo cj6l oo(fiod6l oo('io c;N oo(fJo Ci6l oo (Yto c,N oo +od oq sq @ oo $oq, oodod o.oo J UJa F CI' UI IIJz oN zlI TIJ o F IIJJ o lrJ Jf = o o UIa uI o uJ ulb o UJ uJz2 oo o ulo l.L F 5f ts o. o ll,l E, ll s 6 a o 6lN 6l o F- o o o ro lo o6l (\t o (o(o N l() r* (Y) o(o (f) o c! o o ro N o o o o o o o o o o o o o o dto cn o o o o o o o o o o o o o o o o o o o o o o o o o o o o o utI o o o o o o o o o o o o o o o o o o o o o o o o o o o o o foo o o o o o o o o o o o o o o o o o o o o (o o o o o o o o o m E, qt o o o o o o F s o o o o o 6t o o o o o o @ (o @ F- o (f) o o z =o r rr @ ro CD \t q o, rt r s \r o o o o o o o o o o o o o o o Fo CD (o (o o o o o (o !t to to s sl o o o o o (o @ (o @ F* o ol o o = u.l ts = = B E E 3 = E 3 B 3 3 = 3 uJ t! UJ ]U TU T.lJ ul ul UJ ul IIJ uJ uJ ul Eoz o F - F-I t-I FI F-I t-I FI F-I FI r*I FI t-I l'-I t-I F-I t-(o CJ Fq o Fq o t-q o F-q o Fq o F(o c, oq oq oq r o u? o u? o u? o u? o tr c\l o, F- 6lol r- 6lq F- 6l(D F- 6lq t- C{ CD F- C\lq r- Nq l.* Nq r- Nq t- ol CD F- Nq F- olq F- 6lq F- 6lq t- oo oi oo oi oo oi oo ot oo o; oo o; oo oi o CD CJ o o) C' o CD Ci o CD C; o CD CJ o ct) ci o CD o f EI UJoz UJ @Da N lUoz uJ cofa (r) u.taz uJ cofo $ IUoz TU cof U' (o UJ oz UJdIla t- ]Uoz uJ cola @ uloz ul @la o uJoz ul @fa t.rJoz TUdtla N UJoz UJ cofa lUoz ul(n la 6t T'Joz UIdl :)a (?) t,Joz[! co =a ta, UJoz tU Cnfa (o tIJoz ul d)l U) z o dI C\t z o dt (f) z o dl s z o d) to z o dl (o z o dt F z o d) F dt c{ F dI (f) dt s l- d) (o :< l- dI F- dI @ F dl UId Lll = tr fE. o- f E.(L l E.(L f G(L f E.(L l E.(L ft o- l E.(L l G.(L :) E.(L l E. o- l E.(L l G. o- f E.(L f E.(L o = o = o = o = o = o = o = tr -)oz uJ(L tr -oz UJ(L e. -oz lrJ(L tr -oz[!(L tr -oz tU(L G, -oz[!(L E, '1oz UJ(L ll. ul6 C!o J C\lo J 6to J No J olo J No J 6lo J No J No J No J c\Io J (\lo J No J No J C\lo J oosoo oosoo ooq oo ooq oo oosoo ooq oo oo$oo ooNFo oo6tFo oo C\lFo ooNt-o ooNF o ooNt-o ooNFo =oo dl FJo FJ o FJo l-J o FJ o FJ o FJ o FJ o FJ o FJ o FJo FJ o FJ o FJ o FJ o FJ o FJo l-Jo F -Jo FJ o FJo FJo FJ o FJ o FJ o FJ o FJ o t-) o FJ o uJl- a oodod oo(ciod oodod oodod oo @q to oo(ood oc(oq to oo((,od oodod oo((,od oo Fq o C\l oq i-o CJN oq t-o oN oo F-o C'N oo F-o CiN oodq c\l oo @q c\lo oo(r;o c\io oo(('q No oo @o c.io oo((' o. c!o oodq C\to oo F-o CJo oo l-o (c'o oo F-odo oq r-odo oq r-o((,o oq f-q(oo oq r-q(oo \ (L o o J UIo Fo tu lrJz oN z ul xf ul o uJo IIJ lrJFz o = J o UJ J3 =o o ulo trJ o uJ uJ -o o ul ulzz oo o lrJ o lr. F Jfb(L o IIJ E, t2 o o o o o o os s o o o o o o o |r) N o o o6l o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o l() t- lf) l() (\t o or o o o o (Y) 6l (f) o o o o o o o l.() o o o o o o o o o o s to F to ro (\l o o o o o o o c) 6l t (, l'J T,J lU T,J lU ul UJ ul T'J IU lrl UJ ul TU uJ ul TU tU @ 6,i @ c\i @ 6i @ r e.i @ c.i @ u? Fc! Fc! Fol Fc! r t-c! Fc! otI otI olI NI (\tI olI oq oa oa oa oq t-@d N ro o; 6l r{) oi 6t r() oi (\l rr) o; Nlo o; N l() o; @ o? CD @ o) o; @ CD ot @ o) o; @(D oi @ o) ot fo E,Y ul = 6l :)o G,Y uJ = c) l o E.Y tU = (o fodYtx = t- f o E,Y uJ = coo o-Y ioz so(L oz 5 (f) oz so o- oz 5 ral oz so o- oz 5 N oz so(L l<lalz l5 \r oz soG oz 5 t* oz so o- oz 5 d E (\l tr EF (Y) E, tr s E, trF (o G. F tr l'- G. tr f o E.Y Ul l o G.Y]U = fo E,Y TIJ = fo E.Y tU = lo G,Y UJ = ozo = E. Y E s t }z t $ t $ t $ z E. lUY z E. u,Y z u. tUY z t tUY z E. trJY z E. uJY oo(o =z oo(o =z oo(t) =z oo(Y) =z oo(v) =z ooF-z = ooN E. o oo(\t E.o ooN G. o ooN E. o ooNt o oo C\l E. o oo(\lv o o6tY o 6loAIY o (oo6tv o $o(\lY o roo c.{ :z o G, lUIz E. UJIz E. UIIz E uJIz E. tUIz o z. o = FJ o FJo FJ o l-J o FJ o FJ o FJ o FJ o FJ o ]-J o FJ o FJ o oo F.o cd(\l oq F-od C\l oq F-o cd6l oq F-o .d6l oo F-o cd6l oq r()o C'6l oodo €i oolriq (?) oq lr)o(rt oo(oo o;o oo((,o o;o oodo o;o oo F-q (7) oo F-o ct oo l-o (?i oq t*o €i oo F-o ci oo t-o Gi l3 o o dl oa o o o oo oo o o o o o CO o oooo (o (?)(f) (7' o ^l(ol(ot vl tr) ooooo C\l o oo oo oo o o o o o I;l=1" l" o ol o I oo$oooo oT'J - o ooooo oooo o o o o (oCDooooo oooo(o I loo Io I ;fT"]I o o o I o o ooooooo ol o dlt a o ooooo @ o crl s I o ooo@cno oooooo o @ oFo6l o@o t o o oooooo@N oooooo o o @ ort-o @o(oo I (f) @ or (oto@CDo@ ooooo(o =nJ uJ uJ ulUIuJuJuJ = E 3 = UI UJulTUuJ]'JUJ 3UJuJ E o2 o |r)q 6l lo9 6,1 (\r\ ol 6l\6l to a6l tr) a6l ro a(\l c\l ol o I 6r\6l 6lt-C\l\ c\l ol c! o I 6lNd I (\l6l o a t- c.l N @(7) C' rrI c!roc,l r Fq N6t F-c! 6lGi @ CD o I @ o) o @q o I @q o I @(Y) o @(t) o @(?) CJ f ur J tu G, u.l =tt E ro c! @ N @q (?) N tr o(f) F*flo trJ =ull o r,() c\l @ oo(o f tU =ullo oo cdq t*\ F- t- foYf cn = tU ul =[Ufo oo(o fo UJ =trlflo oo cdq t-\l'- (o fo :z oo cdo @ = IU f Lll =tUl o F-\r- l() f oYf @q s =oY tUf o F-\ F- oo(O2 ul oodo = fo t-\ F- ul o l.Lg(o 26 at (o o fY tU a(o UJq o(?) f-l o ul = o r() l{) c! @ G, o(?) F- f =tu r{) @ t o (o l o(?) f-l LO rr)q E (L Y lo o(7) r- , tU =uJl o lr) (L 6 =fY ul @ g a J o r{) a @ (o $ o rO FJ o @ @ (L 6 lY ul lr) J la @ FJ g o oI J (f) t- J criq rf)(\I oz J ozo = oo6rz = ozo2 @q @ c\r ooNz = oz cdq rr)6r z o = ozo l= @|r) ootf o o @q ol @ ot r.() @q N N l o oosl o trJ =l o oo @ ot <l F uldI ul =uJflo oo cdq C{C! @ oi (t, ul =tu oq @q 6rN (\t l C o; tU oq @o 6iN j o x = oo o,o o r() @ o; lr) fo o oo o d) h') o) (f) FJ o oo o;o ci N 6l l ola J o I E r{) @ o; oo CD ml> t- oo CDq o r() oq CD r f ola oo O)o cdo o$ FJo (o F- s aJJ lJ- os J t-) oo O)o crto (o a)J lJ- fo aF al- No cnl- aIF (o t- $ oo CDo ro (") l- N aJJ tL oo otodo orc\rc!o oodo c.i c\I @q (O c\r cdo <.i N oq @o c.io oo cdq No oq @o lr!l' @q 6lo (L o o .J lrJo l-o ul r,Jz oN z ul xf ul a UJo a ult-f I z ul o UJ J f = o a ulo UJ o ulJ ulb o u.l llJzz oo o I'Jo lr. F J =ts o- o uJ u o o o ooo ooo o o r() =J, @o t4 s 6 o o o o o o o o o o o o I o o F(o C\l o ot (\r o oo 6l olo io dl oo o o o o o o o o o o o o o o o o o o o UII o o o o o o o o o o o o o o o o o o o foo o o o o o t- o o o o o o o o o o o o o dltr o o o o o o o o o to CD o @ t- (D o I o t- o (D @ o o o o o t* tr) o o o rr o o l-oF o F- o CD @ t- o o lo CD t* t- @ F (Y) o o 6l o B UI ul uJ t,J uJ TU TU T'J trj I'J tx ul Lu uJ uJ UJ 3 3 E E Eoz o @q q)q @q? @q @q lot- c, t()l'* ct ro\o F-irl CJ t-|o Ci olI 6lI NI olI r 6lI FI I FI I t-I I trI a EF F@ (c' t*@d F*@(o F@ ((J F-@ (c, @q @ @od @o cd to(Y) o ro(o c; @ ol CD @ o, oi @ CD oi @(t) oi @ CD o; N ol F (\tq t- 6l o) l- 6tq F :) u.l dl o o-Y 6l dt o(L Y (7) co o(LY \r (I) o(L Y (o @o(LY (f, o E, =zF s o E. !zz F ro o G, =ztr (7) oz o t-f oF lo oz o trl oF tr tr tr ol G, EF (7) E. tr tt E, EF to E. tr TIJoz UJdIlo (f, uJoz tU cola s TUoz lIJ 00la ro uloz tudtf U) trl e, EI tr ozo = ozo = oz o = ozo = ozo = 3 s =Yfa =g 3 sfa 3g =$la oFo tro z tr ulY z tr uJY z E, IIJY z E, uJY z G, TTJY ft o- l E. o- lE o- D E.L lt EIo ool'*z = ooi-z = oot-z = ooFz = oot*z = 6l(o J oo(o o oo(oFo ooNY o oo c\l}l o oo(\lY o oo6tY o oo6l!z o 6lo (Yo J No J (\ro J z6o tr! ozo = oz o = oz o = ozo = ozo = oz o = =g =$f,a 3g =g f(n tsg =sfa FJ o FJ o FJ o FJ o FJ o FJ o J o FJ o FJo FJ o uJ o oo o $o oo ci +o oo o $o oo C, Io oq o .(t.o oo CJ ('t oo ci (.J ood (.i oo o l- oo o F- oo ct cd oq o cri oo o cd oo C' CC; oo C, cd oo o (l' oo CJ CJ oo CJ d oo ci d o-o o J UJo Fo tu TIJz oN z LlJ 2 FF =G g lllftr olua z UJ o Lrl J f =oo u.la uJ o IIJJl! b o IJJ lrJzz oo o tuo IL F Jf E(L o lrJt I zl =l lo l5 3. TESTS DE SENSIBILITE OU MESURE DE LA SENSIBILITE Dans le cadre des activit6s de contr6le de la sensibilit6 aux insecticides pour le maintien de l'effrcacit6 de la lutte antivectorielle, des tests ont 6t6 r6alis6s avec les organophosphor6s (Abate, Pyraclofos et Phoxime) qui sont les plus utilis6s parmi les 7 tarviciAes ei vis-ir-vis desquels les larves de simulies peuvent d6velopper plus rapidement la r6sistance. En Zone Est la sensibilit6 avec la Perm6thrine, le Carbosulfan, I'Etofenpros et le Bt Hl4 est bonne d'une manidre g6n6rale la sensibilit6 observ6e vis-i-vis du T6m6phos est bonne ou interm6diaire dans les bassins de l'Oti, Ou6m6 etLac Volta avec des CL 95 allant de 0,378 mg/l i 0,947 mg/I, elle s'est am6lior6e pendant I'ann6e 2000 dans le bassin du Mono otr la C 95 atteignait 1,151 mg/l i Ekpla sur le Sio. Avec le Pyraclofos la sensibilit6 est bonne ou interm6diaire sur tous les points avec la CL95 allant de 0,036 mg/l i 0,051 mg/l. La sensibitit6 avec le Phoxime est trds bonne sur toute l'aire trait6e enZone Est malgr6 la pr6sence de populations simulidiennes peu sensibles sur la Mdkrou (CL95 : 0,064) et la Pendjari (CL95 = 0,052). 4. TRAINING Since the creation of the Onchocerciasis Control Programme (OCP), training has become an integral part of its development in order to produce skilled personnel to carry out its many activities at all levels. - In the year 2000, the East had completed the training of the laboratory assistants of the Operational Bases of Kara, Atakpam6, Bohicon and Parakou. They had all been granted dissection codes and authorized to take part in the routine weekly dissections at all these bases. From 5 May to 15 June 2001, the national entomologist of Ghana received training in atl technical activities of the Vector Conhol Unit at Kar4 Hohoe, Kintampo and Atakpam6. From 16 June to 31 July 2001, the national entomologists of Benin and Togo also received the same training at Kra" Atakpam6, Bohicon and Parakou. Two laboratory auxiliaries (1 from Benin I from Togo) were trained as boatmen in the year 2000 to reinforce ground larviciding treatments of river basins where the entomological and epidemiological results still remain poor in the Zone East. A third boatman from Ghana will be trained in 2001. Two technician entomologists from Equitorial Guinea (Malabo) received nvo weeks training in larval prospections and ground larviciding treatments at Kara in the course of the year 2000. l5 7. OTHER TECHNICAL REPORTS In January 2002 the senior author of this texte wrote two other technical reports. These were: i) Laboratory rearing of the various members of the S. damnosum complex in the Eastern Zone of the Onchocercias Control Programme in West Africa 2000 and 2001. ii) Research contribution for the JPC 2001 : Zone East. 8. MEETINGS - The senior author participated in the meeting on Operational Research and Stategy in Ouagadougou from 9 to 1l April 2001. - Mr Yaya Coulibaly and the senior author participated in the Annual ZonalVCU Meeting at Bobo-Dioulasso from22 to 26 October 2001. 9. OBSERVATIONS / COMMENTS - Six cytospecies of the S. damnosum complex (S. squamosum, S. yatrense, S. damnosum (s.s.), S. siiUanum, S. sanctipauli (s.s.), and the Beffa form of S. soubrense were recorded in the course of the year 2001. - As already signalled in the 1998, 1999 and 2000 reports, more cases of the backcross (hybrid) beiween S. damnosum (s.s.) and S. sirbanum were again registered in the year 2001. - The hybrid cases were recorded on the following rivers in the Zone East : the Pru, Kara, K6ran, Oudm6, Daka and the White Volta at Sugu in Ghana. 10. ACKNOWLEDGMENT We are sincerely grateful to alt members of the Vector Control Unit (VCU) and in particular to Dr L. YAMEOGO (Chief VCU) ; Dr L.K.B. Akpoboua (ATOA/CU) ; Dr H' Agoua (CAT Zone East) ; Mr M. Sarr (CAT Zone West) ; Dr Y. Bissan (CRE) ; Mr Ake Assi (Coordinator/Aerops) ; the Chiefs of Operational Bases and their assistants as well as the entire staffof the two aerobases at Kara (Togo) and Odiennd (C6te d'Ivoire) including all the Evergreen personnel who had provided materials or assisted in one way or the other to make this work a success. 50 rrt c) F.t -, -{ E -{ E(J FI F c)o =toa - -{ 5z E' rrt tn Hr, FI c) tn ?n 4 a -lt rnz ,{ E6) -16za o 5o -1 xozo =-o ttt tn l: o C)t t 50 EFa9z zz Erl rrt NOoo JJ o Doo D2 o a = F o E DC{ 2 -a llt7 o s -lll m E D2 -I - -o m v 2 -ol[ v - D E Dn .ll F Clll -{o =6lm7 no{ oI! Da D o C llt - llt F Dn .ll .ll FCil -{o - o - o F D .ll nnFC n2{o DnnFC n -{o F D o oF{ D lllo{ DnnFCllt -{o F D o oF{ D o C mo{ - o F nnFClll2 .{ o Illt a - F DnnF m - -{ o I Da D ec DnnFClll -{o oj ! 1 - a! D? 1 I m v m o Doo D -I a D = F o - -o n v o _c z lltllt E D o CFlt - -6l llt v o C llto{ 2 -oI[ v llto{ o C n - m o Clt - lll - o2 o so oF{ D oF{ D o?{ D oF .{ D a oF{ D oF{ D E Do 2 - I(,l N 6l ol Noo (,lj tlN j 5o I Llil N(,l 5 No{ I ul{ 5(rl oN I oN I5o N5(,l { o{ DF o{ tgl Not o (.1 o (,lN INgl j o(,l No o o (, rl J tr D -2 o (,o Ngr{ 5I (.1 I5 ul ogl o{ NI gl 5tl (,lgl I5{ I(., t,| N5N I vE NN o o o o o o o I I o t o(o o o o o o a oC D o o o o o o o o o I(rl o o o o o DElll I(o o o o o o o o o o o o o o o o Doo o o o o o o o oj o o o o o o o o oE D I(o o o N No o N I ut 5 o gt N gl N o P t2 @ c2 z or{ o! mI mo oo =!o @4<=qo =2#o r.ll tsqa= =L'1- b836 o m{ mv ==I oz ch o oit oN 6 6 at oN ct oN ct N ct oi! clit Ct N CIN o 5 ctiJ cl 6' ct o 5 ctN cl ol cl cl (., C!IS ct G' CIN CI (., CtN Ct (, oN o ot a N o o!o CI ct Nit oN cl NN CIit o o(o o ct N NoN CI oit ct o o ro Ct Ct N oit ct 5 o ot ct 5 CIg, Ct NN oi! C' NIS clN o NN ctN o o ro o c) ao 6 ct rO ct cl !o CI o to o ct Nql o o (,t o o ot ct ct c, m ! a m =m 2o mo LocltD ol .,l PNo5 No o 5 ov5 N I! m oC o 5 ov5CIo x a o 5 ov5oo x v oI ouotoo x v o 5 ovNoo x 7 o ovN6o x v o ovNCIo x a o 5 ovNoo I v o ovNoo I v o o7Noo F7 o o7No6 x v o ovNoo x v or ovNoo x v o o7Noo x 7 o o 6l xoCxoC3 @oC o o (Doo oI o o (Doo oI o o d)oo oI o o (r!oo o! oI o altoo oI oq o (,too o = o o oloo o = o o 6loo o = o o 6!oo o = o o 6!oo o = o o (,loo o = o o 6lo6 o = oI o C!oo oj< =@ o o 6loo oI =@ o o (D6o oI m . o o (D6o o = o m o o (Doo oj o 5 o oroo ! mzo 4. o o Noo ! m =o 4, o o Noo ! mzo 4, o o N6o ! m =o v oI o Noo ! m2o 7 v m @ t, t2 z @ m1l v s m7 m ?I x1' m oc t2 x z II 2 o ?x = @oco @oco @oCo o =o o2 o Ezo !o N ot m m F2o !o N 5 m m E2o !o N ot mt- m E2o !o N N m m E2o !oN - m m xocxoc =@oC oj @ mo o - z o o - 2 o oI !oz{ ,oc =mv o =!oz voC =mv oI !o = voc =mv oC =o2 o oC =o =o ot mt- m ocI o =o m m s ocI ozo 5 m m s oC =ozo al m m ocIozo N m m s ocI o =o oc =ozo ocj oz o oCI o =o o c, =o2o !ovo @ j 5 m m s @ = o, m m s @ j N m m @ = m m tr mC !a6t5 @leto (o\ol ot ol6t ao\6r6t Or6t @ '6r 6t(D6t Jo ot ol6t -@(,t (D 6r Jo(,r ool Ja or(,t(.t ol6 Jo(,r(,(.1(,t(.t -ao(,l o cr Jogr arr6t Jo or 6r(D Jo or (D ol Jo ol ot .D P olooo I ot6to6t oL6(.,(,t 5 oLo olG'5 Ig, ol aD ol P(., (D 6! ol PG' o6r(,l P GT orooo P ol(,rooo .o(, otoocl Pq,(,looC! -oo,qlo6 ct IG'(,rooCI P6t(,loocl P(,, orooct P(,,(,rooct .oG'gr o €lct -oo, anooa oaolol6t -o(oaoo o @ooo .P @oocl P(oooo 5 = =o o 'ro 5 rO ao ro m P6 oloo m .9cl otCIo m P(o 6! ol ot m i$ql(., o,5 m irrgl o,ql5 m is ololo! m .N oooo m i\toooo m i., ol ar! ot m i\! ar! ot(r! m i., ol(,l 6) m 'N(D ot6) m i.ro o!o m Pgl ogl ol a.l m P ol6o6o m -oolqloo m .P(,lgroo m P56ro(,r ol m .P56!(rr 6!(D m oLolocto m P ot ol(r! ol .D m -o(,r Oto O)o m P ol6t(rr6l(D m -oot ot ar!o6r m P ot(D 6to6l m -6gt 6l ol6) ol m Pgl 6l(D 6l(D m .Pgr 6! ol6l o! m -oor(,l ol ot ol m _o(,r(,r olct6l m .9 orC'6l olol m P(o 6ro(D m 'or oooo m inooo6 m i,oooo m irroooo m N Na.r (,to 4., 5 6 6 N o o N o A o N6 ('r ('r N6 N 6 or N 5 5 5 ('r Nor olo (D cl a, o ct a, 5 N N No 19o! 4., 5 (D a, N 6 6 N CD 5 o N ot (,l No N 6 ol N 5 5 5 ol N5 N@ 6t ct o 6 6 - m t-5tlollE,llol I url lml totl2l lql HIl=lt-ll-{ltot lElt-JIolt>tl=ll2lt-Jl.,,l lEl lalol sl t(o_1t>t I Urll anl Itt lloll@ll>l ;IJJlAl E IEI sl\lcrlol4lf I ID 9,z o U)t mo ma o o =!o @2<=TO =2#o 711 sq@a = lt,z,- "88 36 o m mv =z =oza \0 o{ o(,, o o{ o(,, o o{ o o, o o5 o c) o(.) o o o(, o o (.)o o o o(, o o (.)o o o o)o o o o)o o o) o (), o o(, o (, o o(, o (., o o(,, o o,o o G' o N Or o(o o N or o @ o N Or o(o o 5 oN o 5 oN o 5 oN o 5 oN o 5 oN o \l o o @ o o { o o { o o { o o { o o o(.' o o o(., o o o(o o o oN o N) o 0{ m ! 7 z U' mo -{ Lo @ @(,,(, t-ooo o @ m ot-{ 5o o - 5o o 1- t- 5o or{ t- 5o or -{ t- N{ ot-{ t- Ng) ot-{ r NN or -{ |- NN ot- -.1 t- or o|- -{ Ol ot- -{ @ a0oo ot-{ ID(ooc, ot- -{ ID(ooc, ot- -.1 @(O oo ot-{ tr(ooo =t! ? or -.1 oxG'No -{ m = ?or -{ oxG)No o -.1 m or{ ox(, No ol- -.1 oxG)oo ot- -{ oxNoo ot-{ oxNo cl ot- -t oxNoo o|.{ oxNoo or{ oxNoo or -{ oxNoo o -{ oo Otoo ?or -{ oo5oo ot- -.1 oo5oo ?or -{ o o5oo ?ot- -{ oo5oo ?ot-{ oo5oo ?or -{ oo(r,oo or+ oo o o{ oo o or .l o n Otoo r 7 m E U, 9.z @ ffi .Tt v,{ a{ oC U'{ tn{ oc U'{ tn{ oC 0 ECt-c IDoz o oI tr ct,(n @ a tn o x o x {I -.1 m or -{ = + =m ot- -{ = t =m ot- -{ = I =m ot-{ E -; m or{ -{ xoC mz -.1 { xoC mz -{ { xoC mz{ x mv z x ma z x mv z x m7 z x mT z x m7 z x m7 z =o =o =o =o =o =o =o (D oc r m IDoC r m x v n m7 m o -{ (tt -{ oC 1t t-t- Ut or m!- @ -.1 (t, i oC .Tt t- a G' m!- U' -{ a{ oC 'Tt rr(r, m!- € mo ID m -{C -lcx! mz m x moI m tD x! U'(r, x! a at, mxCzo ! m N mt- m mxczo ! m mr m U'C oc OI mr m U,C oc s mt- m U,Co C G' mt- m (n C o C N m - m s U'Coc mr m xoCz{ oc o oC o 7 xoCz{ oC o oC (D v xocz -{oCo oC (D ? ox o,oo -{ =v 3 o = =n or mt- m = =, 5 mt- m =I7 G) mt- m = =7 N mt- m = =n mr m xoz q, o z tr o z or m|. m (D o z 5 m m (D o z(., mt- m ID o z N m -m =o t-m + mng m t- m + mn o m 0(- u, oz oz tr mC { (.) G'(., G) .\t (.)(., 4.,(., _{(,(,(.,(, -{o(o @(o(l' -@ oooo -o @ G)(.,(.) -@or o,(r, o) -ooor (D o)Ol -@t(o(o(o @L5(o(o @ -(o CD oroo -@@qloo (o to Oloo -(o CD Oloo -(0o Oroo -o o) o) o)o -o (t) o)(no -o CD(D(,)(D -o C')(n o)(n (o -@ @ G) G' G) -(0(oo(.,(.)(.) Jo(o CD(.,(.)(., -(o(oo(,(.,(,, -@(oo G) G'G' @ io @(.,(,(., @\o5{ G) -(oooooo -(oooooo -(oooooc, -(ooooocl Joooooo -(o ooo -(oN (n CD{ -(oN o)(n{ -(o co @(., G'5 E -{ -{ o P(.,o(,(.t G' -oG' o G'(., G) -o(.,o(.,q,(., -o5G' a.)(.,5 o .G' oo(., o) GI -o(., 6 o,(., G' -o(., CD(., G'6 -o(., @(.,(.,(.) -oN o)oi{ IN ol <,,{ oio @ o, a.)(, -o(o @(.,(.,(., _o(o @(.,g) GI -o(o @(,, G'(,, -o(o @(,(,(., -oo) o,Olo _o(D o) C')o o b, (n o)o o -o! (t) o)(D o)(D{ L (n(r,{ '- o) o){ (Do){ :r (n(n{ o)o){ oi,5oo o -o, (D o) o){ I o)(D o)(t){ -oo) o)(D o,{ -o(n ct,(n ct,{ -oo) o,(n o){ b orooo ie o) o) o) i.) o) oi o) P Ol o)(n C" C') t-oz o -.16 m m m m m m m m { = € = € = { m m m m m m m m m m m m m m m m m m m m m € 5 o Ol (.) (., (o @ g) (o 6 N G's (., o) o,N @ (.' o N o o (n o 5 o E or 5 G' -{o{ r Or N N G) @ 0 =z (o @ o) (o o N (.)5 (.) s (.)N o (D o N o G) o (n @ s o 5 qt 5 G' tt, n ID N U,oC 5 @ Tm @ oa a o TD 5 oN o A oN o 5 oo o o o o J NN o OI o NN o OI o NN o Or o NN o or o N 19 o or o NN o(., o o -{ m N{ o N) o o o o (t) o o (n o o o) o o o) o o oN o o o(t) o(, o o(n o (.,o o o { o o oN o o N ot o5 o N Ol o5 o N ot o5 o N Or o5 o N Or os o (n o or o @ o N o o cn o N o (.,o o o o(o o o o N o o z v, mo{ I m =m -t m3 m INo5 No PNo5 No t-oo c,o @ m o s o =u, m = o -{ m o H z =z{o OI =z{o Or =z{o Or =z{o OI =z{o OI =z{oqr =z{oOr =z{oor =z{o =z{oo =z{oo =z{eo =z{eo =z{oo =z{oo =z\.1oo =z{oo =z{oo =z{oo =z{oo =z{oo =z{oo =z{oo =z(.)o =z(.)o =zNoo =zNoo =zNoo =zNoc, = zNoo =zNoo @ m 'Tt U' o aD o (tt 6 at 6 U' 6 9,o att o 9,o =oC ooxo o oxo =oz o =oz o =oz o =oz o =oz o =oz o =oz o =oz o =oz o =oz o =oz o =oz o oC oc =oz o =oz o =oz o =oz o =o =o =oz o 4, m n m mx! E mx! E mx!t- mx!t mx!r mx! E x J =oz oCo ID =oc o tDt-o Eox o =m t! n7' o m 0oxo =m ID vv o m x! m|- mN x! o TD OT mr m x!o(D 5 mr m x! o(D (, mr m x! o TD N mt- m x!o(D m -m 0(- v t- 0(- v F 0(- v t- o(- v F oL T E o(- v F oC -o tn U'o oc t-o U, U'o t- z o =oz o or mr m r z o =oz o 5 m|- m - z o =oz o(.) mt- m t- zo =ozo N m m Ezo =ozo mr m Ezo =oz o mr m tr mC o)iooooo (,) -ro oooo -o)(o oooo I(ooooo (t iooooo -ot(o oooo -(nCI, @(.,(.,5 o,\ o) ctr o) CD -o@ G)(.) G' G) -o,(o(,,(,) a.)5 -o)(0(.,(.)(.'5 -o)(o o) o)6) -o)@ o)(D o)6) I @ CD o) C')6) -(n@ CD o) oi6) -o)@ o)(D(n 6) -o)@ CD o) o) o) p(ooooo (n iooooo o) -@ oooo -o)r0oooo -o)(0oooo (n iooooo !o{5(o ro(o @\5(o(o(o @il@(,, o,5 @ -Ol @(,(,5 -6Olo(.,(.,5 !o Oro(.,(.,5 -@grq)(,(,)5 P Or@ o,(.,5 r IIo P6 6) ct,{ o to C') o){ -oo o, o){ -o@ o) o){ I @ o, o){ -oqt o) CD\t -o@ o) o){ oiooooo b OIooo -o, C') o) o)6) -o, (n o)(t) 6) -(,r @(.,(., o -or @G'(., G) irr @(.,(, 6 -(,r @(l)(., o or@G'(., o -Or @ C.,(, 6 -ot ooo -o ooo -o ooo -ol ooo b,ooo b,ooo -o OIooo -o (rtooo ie5(o(o(o ie5(o(o(o ie5(o(o(o ie5(o(o @ ie5(o(o(o i.,5(o(o(o t- oz o -{6 m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m{ (,, or A N6 N){ Or 5 N G' o) o (,t @ (,I o (o NOr N 5 6 (,5 @ J 6 o o o { 5 -to -{ t- N (., 5 { (., j (.) ct o or @ (,l o (o NOI N 5 o (.,5 o =z N o $ U' n !0 N { 6 5 (., N @ o o { aoE + m @ @ U,(t o ID E EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE E :i, t : .. I s.= .1-" . = ,,.; it :::. i I a. lllIrlrIIItll lrIllllIlll rIlllilIllllillllll Itlll @ a2 =oz o o! mo rII U, o o =!o trE<=v9 H5 v11 @Q>acn= = (-, 1- "ts8 =o o m mv = =I ozo No @Iz acx =x = @!lilo EI U, oo = .o o @2<=TO =7#on'n sqa= =L,z,- L836 o m In n _=z =oz U, N o5 o o o o o o(., o o o(., o o N o) o o N(.) o o NG' o 0 N(., o o 6 o o No o5 o No o5 o No o5 o No o5 o 0 -lm =@ mo+ ! a -{ m =m Lo @ @(, G -oNo5 No roooo U, m Fn m @ U' 9.z t U'C x €x FaCx =x Ea L x =x FaC x{x =I! acx €x =ID acx €x H -{ m = o -{ m E E E r G'{ t-(.,5 rq,5 r(.)5 (.,5 r(,5 t-q,5 -(.,N t-G)N rG'N -(.,N CD m 'Tl |-(, @ t- G){ U'Cx €x{ @Cx =x{ U'cx €x € 7 m n m =mzC o ID z -oC o(D z I oq otr z -oC o ID z -oC otr z I oc otr z + oC CNCx €x € =mzC @ o I =zaC I =zaC o TD o(- oo! o(- o0g o(- ogg o(- oo = 0(- oo! =zx vo Or m -m =zx 7o 5 m m =zx vo(.) mt- m Izx vo N mr m C, mC -{{ q) CD{ { b,o(,(.)5 -{(,) @(o @@ -{o)(o(o(o 6 -{ ct,(o(0(o 6 -{ ct)(o(o(o @ \l b,(o(o(oo ob @(.,(,, 5 6b co G'(.,5 -@o@ o, GO5 -@o6(.t G'5 E J-l J...lo -{ot5 @ @ @ \l\ o, cl){ -o{ C'Iooo -o{(,ooo o\ Otooo rozo -{6 oL @ @ro(o -o or Oloo -oor(Jroo o\ @(.,(,5 -o or @(.,(,5 -oOI@(., G'5 -oor@ G'(.,5 -o or @(.t(,5 -oOI@(., G'5 o\(,tooo m m m m{m m m m m m m m m m OI 5 5 o (.) 5 N$ (.)o N)or 5 (, 5 -{o -{ r 0 =z(., 5 o) @ (.) CN u TD (,r 5 $ o @ (o N(, 5 (.) s U'oc I m (r, @(r, @o @ illllllllllllll fl.:il ilI HHEE li:\ i r:: i @ a z oc m =m o! mo nIa o o =!o @2<= -o #o n'rl Bq@A zF "88 36 o m mv = = =oza NN o5 o o (n o ot o (D o OI o (D o ot o o) o OT o o o(, o o(., o o N oN o o(., o o (D oN o { o Or o { o(,l o { o(rt o { o or o o o o (o o or o (0 o OI o (o c, or o (o o OI o (D o ot o o(.) o o (o o(., o (l, o o, o (o o(, o (o o(.) o (o oG' o @ o(., o 0 -.1 m ! v -{ m =m =@ mo -.1 Loo(o(.)(r! -oNo5 No t-oo oo U, m 6c m =m 6-q m =m oC m =m oC m =m oc m =m oC m =m oC m =m oC m =m oC m =m oc m =m =ID oC m =m =@ oc m =m { m = oc m =m o{ m oC m =m 6-E m =m oc m =m oC m =m oC m =m 6c m =m 6-q m =m 6',c m =m 6c m =m 6C m =m oc m =m oC m =m oC m =m 6',C m =m r n m E U'2z oC\l5o ox! n oc{(.)o ox! n oc{(.,o o x! n oc{(.,o ox! 7 oC{G'o ox! 7 oc{(.,o ox! 7 oC{No o x! v oC{ o {oc U' @ oc{ o -{oc U' 9, oC @oo oc m =m oC{oo oq m =m oC{oo oc m =m oC{oo oc m =m oC{oo oc m =m oC{oo oC m =m oC o,oo oC m =m oc(noo oC m =m oE o)oo oC m =m oC o)oo oC m =m oC o)oo oC m =m oC o)oo oC m =m oc5oo oC m =m oC5oo oc m =m oC5oo oc m =m oC5oo oc m =m oC5oo oC m =m oC5oo oC m =m @ m .Tt v mv m o(- tr{ x TDoE ot mt- m x TD o 5 mt- m x TD oC (., mr m x ID oC N mr m x (D oC mr m TD @ U' @ 7v o m = m 4, tr vn o m € m7 tp vv o m o(D o o trz ! I oC o o tr ='o I oC o o trz t I oC oo trz ! I oC oo trz ! I oc =trm{ mxocx oc(,t m|- m = (D m{ mx oCxoq 5 mr m = (D m -{ mxoC xoc (^) mt- m =trm -{ mx oc xo C N mt- m = (D m -{ mxocxoC m -m x! x! 6 TD o ID m{ mn oC{ mt- m (D m -.1 m uoc OI m m (D m{ mv oC 5 m m TD m -{ mvoc(,, ml- m u, m -{ m7oc N m -m q, m -{ m noc mt- m tr mc -{aoooo -(oN Oroo _(oN Oloo -(oN OIoo -(oi,(,oo -@N(rtoo -(ON co o)G'5 Jo(,)(,)(,(,5 (oi,G' o, G'I o, io5 ao @(o -{o)(.,(.,(,5 { brG' G'(.)5 { b,(,(.,o5 { 'o(, 4.,(, 5 -{(t)(,)(.,(.,5 -{\l o) o) o)o {\ ct(n(n(D \l\ o)(t) Ol ct) .\l{ o) o) o) C', -{{ o)(ng) ct, -{(l' o, o) o) -ro @(.)(.)5 -'o o(,(.,5 -(g 6 G)G'5 -@ CD(.,(^,5 -(o o(),(.,5 -(o o(,,(,5 t- = =o N -o, OIoo N{ gr o)(D N\t o)(D o, .I'{ o)g) o) !e{ c)(D(D J\,{ o) o) C,) N\ G)(.,(.)5 N\(n(ng){ N\ o) CD ot{ NL(,(.,(,5 NL(D(D(r,{ NL o) o) o){ N5 o, o) o){ N5g)(D(D{ NL(n(ng){ NL5 al, @ ao N55(o(o @ N55 ro(o(o l\,L5 ao(o(o NL5@(o @ J.,(.) o) o) o) Ot NN(D(n(n ot NN CD ot(D C" !.,N o) o) o) o) !eN C,) o)(n ot NNg)(D(D(n NN o)(n o, ot 1-oz o{ 6 m m m m m m m m m m m m m m m m m m m m m m m m m m m m{ o o O) 5 { (.,OI Oro 5 N5 5 { Ot5 (,,$ G'G' o o (,r N (,) { -.t { o o o (o -to -{ t- o o o) s { G 5 (., 5 N5 o No o o (,l N (., (., o =z N5 5Ol 5 o N 5 { { o @ o (0 U' v @ aoC T m o @ @ G' (., o N5 (J U' o u, ( ?i.: m @9, =z6 mv mo @ 9,z z 6 m n =tr CJ'! mo ma o o =!otrq<=vo H5 v11 sqaa =L)z,f "88 36 g m{ mv =z{ ozo a! m o ma o o =!otrq<= ,o9. fr5 v11EQ>aaa =LJz,- "ts8 36 o m mv =z J oz U' N(, o -.t m (o o\l o (o o{ o (I, o{ o (o o{ o (o o{ o (o o{ o (o o{ o (o o{ o N(o o or o N(o o or o N(0 o Or o N(o oqI o N(I, o Or o N(o o Ol o N(o o OI o o(o o5 o o(o o5 o 2 @ mo -.1 ro o0 o U, m t- v m z 6 m7 z o m7 @ CI'Iz z 6 mv z o ma z 6 m7 =6 mv z 6 m7 z 6 m a z o mT z o mv z 6 m7 z 6 m n z 6 mv z 6 mv z o m7 =o mv z 6 mv z oroo z (,Ioo z oroo z oroo z oroo z Otoo z otoo z (r! oo z =(.,oo z =G'oo z =(.,oo z =o,oo z =(.,oo z =(,oo z =(l'oo z U,Noo z @Noo E m 'Tt @ o -.1 7 m7 m tr(D ov trt! ov trI! o v tr IDo7 trt! ov tr TD o7 tr TE o 4, tr @ o7 =mx7oC =mx noE =mx7oC =mxvoC =mx7oC =mxvoC =mxnoC U' o -{ tr TD ov (,) mr m trI! o7 mr m tr oo7 (o mr m tr(D o n o mr m tr(! ov{ mr m tr @ o n o) mt- m tr ID o v 5 mr m tr ID ov mt- m x mv m =oC{ mt- m x m n m =oC o) mr m x m n m =o C Ol mr m x mv m =oc 5 mr m x m7 m =oC(.) m|- m x m n m =oc N) mr m x m7 m =oc mr m o TD @(rt m OI mt- m o TD U'a m 5 mr m tr mC i.) @ @ @@ ie(o(o(oo i.,(o(o(o @ i\) @(0(o @ ie(o(o(o 6 irl(o(o(o(o .N (o(o(o(o irl(I,(o rI) @ Loooo boooo -5 oooo Loooo Loooo Loooo Loooo -@ OIoo -@ Oroo F = =o N '@ o) o) o) O) N '6 C') o) o)g) N tog,(D(ng) Niog) o) o) C') N to CD(Dg) o) !s @ o)(,) o)Ol N @(D o)(n 6) N @(n(D o)o J.' @(.,(.) 6 J\' €D(.)(., 6 !., @(.,(.) 6 !., @(,(., 6 N @ o,(., 6 !., @g, a.)o Ju @(,,(.) o (.,b5(o(o @ G'b5(o @@ |-oz o + 6 m m m m m m m m m m m m m m m m m m = N o 6 6 o o o o N CE N { A 5 (o or o { o -{ t- o =z N (n @ CD o o o o N @ N { 5 5 (o (,r o Lv@ o oc I m v, @ @ U' o ID ffiffi *$lK f":.n $H]$ s-t\_\ i:,:= lii iii ilffi llllill II I IlIIITlltlll E E H oboooo o -ol N Oroo t-ooo otn m E H z z 6 mv =tr z 6 m u =tr E E E H - = =o t-ozo -{6 € € m = (, -{o -{ t- N@ o =z (., Iv TD @oC I m a @ U, @otr lt HH HH E EIE E E E :t. I ffiE tiffi .: .r.,'ti wfi I |:: l,"t :,:::x.j f _.tr: i tlIl ITll EE EE EE t I (D L =(D z Iz 6 rfl7 ID L =J- C z 6 rfl v oC z m m a! rflo m @ o o =!o @2<=;o #ov'n oQ>a9,4z= "ts8 36 o m ra n =z =oz U, @! m o ma o o =!o s4<= vO H5 n'Tl oQ>-9,4zF "88 36 g rfl m7 =z -{ ozo N5 N o{ e Nq, o o) o N o OI o N o) cl5 o NN o GI o 0 m I -.1 m o -{ m o{ m z aD mo -{ -oooooo o -o oooo o -o oooo oboooo Iooooo t-oooo @ lTl t- v m @ z z 6 m IDoG'oo @ oct- m tD z z 6 m TD oG'oo TD oc -m (D z z 6 m @ o(.,oo ID oct- m (D z z 6 m tro G)oo tD oct- m (D =z 6 m troG'oo @ oCr m tr a 9,z ID fr 'Tl v =?m7 m 3 o z o U'a =o z o @ @ =0 z 0 U' @ =o z o (r, @ =o z 0 @ U, tr mC -o@(., G' o,6 -o@(.,(,, G)o -oo(., G)(., o -o@(.,(,,(., 6 -oct(.,(.t ot6 E = =o -{o)G'(., G'(., = -{o) G) G'(.)(,, € -{ot(,(, ar,(, = -{o) G'(., G) G' € -{otG'(., G) G' = oz o -{6 m{ @ { N o o -{o -{ t- 5 o 6 0 =z 5 { (o G' o 2,n@ U'oc I m (rt U,a U' otr N(D o CD o G)o o or o N{ o5 o x, @ o{ o o{ o o) o o o(,l o o o OI o I{ m o{ m o -.1 m o -{ m o =m o{ m z U, mo{ -oooooo -oooooo -oooooo -oooooo -ooooo cl s -oooooo -oooooo roo oo U, m t- v m + C{ z 6 m = .Tt Noo + c -{ z 6 m = .Tt Noo I -{z 6 m = .Tt Noo I C{ z o m Iz(.,oo I c -lz 6 m -z(.,oo I C{ z 6 m rz(,,oo I C{ z d m TzNoo o @ 9,z I! m -ll = .Tt oC = 'Tt oC = 'tt oc z o mv o sz m m 6 ov =ov m z o m7 oC z m m 6 o7 =ov m z o mv og z m m 6 o n =o7 m z o m7 oc z m m =tro T = mv m tr mC (o\@ G)(., G) -o(., @(,(t(, € -(o\t @ G) ol G' -o(,qt(,,(.,(.,{ -@{6 G' G)(.) I(.) @(,, a.)(,, € I5G'(., G)(., oL(n(n(n \.1 = ob(.,(,(.t G' -o5(Dg)(n{ € o '5 G)(., 00 G) -o5 o)(n C"{ € P (rt o) ol(D -o{ o) C,) o) o) € r JJo t- oz @ =o m € N(.) No) (,,o G'6 o No o io{ t- N OJ o 3z N(n (.)o (.,ot (D Nor or U, 7(D ooC r m U' U, U' CI' o ID N(., o(t) o N G) o o) o N o o) o N o o) o N o(,) o N o C') o No o o) o No o(D o No o o) o No o o) o o -{ m :z @ mo -{ E E 5 P P E H (p m U' @ B E E z E z o oroooc o oC ! m ID tr 1' m ID tr (D o a m = mz Ip o a m =mz TD o @ m =mz q, o U' m =mz 6 m U'o o 6 m @o o 6 m U,o o 6 m U'oo tr mC -@(.,o G)G'(., {i.l o, o) o){ -{N(D o)(D{ -{o(D o, o){ -{o o) o) o){ _{o cr,(n o){ {b o)(n o)\t _{(o(o @(o(0 \t io(o @(o(0 j.l(o(o(o(o(o -{(o(o(0(o(o t- = _{o -{o)G'(.,(.)(.) j.| o @(., 4.,(.t -{o CD 4.,(r,(.) -{{ C" o){ -{{ (D cr,{ \.1\ g) o){ --.1{ o) o){ .\to@(.,(.,A -{o oo(.,(.,5 -{o CD G'(.)5 -{o @(,,q, 5 t-oz o + 6 = = = = = = = = = € € m = N or (,l ao NN N5 N(., N)(o N -{o{ 1- G) 6 6 { N N@ N o =z N (r, 7 @ NN (t, oC + m I o g) U, o @ U' otr IEIEEIIIIEE EEHHEHHEHEE ffNNIffiIEII HEHHHHHHHHg HEHEHEHHHHH E E E :. l i : i;!;' I .:. aa::ii lr'., , iil $if4 :rl i,1,,.' :i :N : . ,::;l ruE @ I = =o @ Lz a aa z0v @! mo ma o o =!o @9<=no =>#o z'11EQ>-cna =(Jz,- "ts8 36 o m{ mv =z{ oz @ a! mo rflo o o =!o @2<= v9 H5 v'lt sqoA -="88 =o o m{ rTI v =z{ oza N No o{ o (D o OI o (., o5 o o{ m o =m o -{ m z U, mo{ o -o oooo oboooo Iooooo m =t-o =t-5eo = t-o =t-5oo =ro =t-$oo a =o n o(D mg oc oo L =ro =-o E o7 o @ m0oC ooc Eov o(! m0oc ooc = -o E H m o oooo -o oooo -o oooo E = =o (oL G)(.,(., G) = (oLG'(.) G)G' € -@5(,(.,(.,(., = E H N N)o NN -{o -{ t- (.) 0 =z qt No NN 9,v @ @oC + m a an(n U' o u, H E E E E E II il,i l:l j ll IT l *m i i.i.l I"1 : ::r. - l i::A ! I tr ltt = =I o D{ I o zo o { o{ Dx o z o = I o C lrlo D z =lltlll Noot !fr r: oo! t - e 'aey- a TN.t\t l-i7'N o o 5S 8'g 8P99. oS U' ffiwmmEE @<ac)ao >>,o>=>3ig''q.dL (^,(r(roooo .i i \) TI h, u. ,...., m x .Uz 8sr C N o\ { t (n. 'tzz 16 r-s9