Final report: vectoral role if Simulium damnosum complex species in Togo and Benin

f - i' Jôrg GRTINEWALD r z/tz//6 lisu02AA fa:;f lnt,,,,',- 0ctober 1985 Ë.4'j o- Final v'n vEcroRrAL -ndr.e or srMulrt M DAl.{NosttH cu.{plEx sPEcrEs rN roco AND BENIN (Synthesis of the reports of various consultants) Precise infornation on the vectorial role of the vector species in Togo and Benin would be invaluable in planning Èhe aerial spraying operation to conErol Slnulium in the Southern Extension area. That also applies to betÈer ffindingthesignificanceoftheATP/ABRva1uesinre1at,iontoEhe epideoiology of onchocerciasis. Accordingly the following reports and documents have been analysed: Barbiero(1982), Cheke (1983, 1984), Cheke & Garms (1983), Garms eÈ al. (L979, 1980, 1982), Garms & Cheke (in press), Ourar (1982) Renz (1982'1983). The comparison of the results of these documents is hampered by several facts: - The main subject of the studies carried out by the various consultants ras not only Èhe vecÈorial role of the vector species concerned buE also reinvasion and expertnenÈaI treatnents, species differentiaEion, dispersal, experimental infections, and idendification of animal Onchocerca species found nithin Èhe vectors. - Therefore the various consultants calculated and presented their resulEs in differenÈ ways. Ihe catching poinÈs for which detailed data is given vary; !tsR/lffP values are rarely presented and are nearly impossible to calculate from the daÈa given; the ratio nulliparous/parous flies is often unknown; furthermore one consultant calculates L3/10@ parous females, the oEher L3/1000 biting feoales. - All studies were carrled out in the rainy season, occasionalLy extenting into the beginning of the dry season (Barbiero 1982,0nar 1982, Renz 1982). Dry season data, however, based on a detailed species differentiation is missing. Entomological evaluation data collected by VCU identifies the various Yect,or species by the ving tuft colours only. Since they vary in Togo and Benin from dark to pale within.S,=_ggg!legsS/§=__sa4ggjggli and §._qq-mmosum_ they are not, reliable for the idencification of the vectorial role of those species. fire occurrence of the different vector species and their respectlve contribution to reinvasion is vell documented (see reports Eentioned above, besides tJalsh 1984, Baldry 1984) and partly published (Garms et al. 1982, Cheke & Garms 1983' Garns & Cheke, in press). Hovever, the Èransmission of 0. volvulus- by the various vectors in dlfferent blo{limatic zones of Togo and Benin is not fully understood: - to what extent contribuEes each vecEor species to the transmisslon? 7 aI 2 - to vhaE exÈent varies Èhe vectorial roLe of the various Yector speciesin different bio-climatic zones? In order to compare the various daÈa collected by the consulÈants it was necessary to recalculaEe then on a coEnon base: TABLE.I shoys this data for 12 catching points from Gbassé in Èhe north to Tététou in the souÈh. The data given is: - the total number of flies dissected by each consultant for each site, - Èhe proportion of each vector species at a Siven site (species composition), - the Transmission Potent.ial (T.P.= number infective females x mean number of L3 per female), - the proporrion of the Transmission PoÈential of each vecÈor species at each site, - the mean number of lii per 1000 parous females (it would make more sense to calculaÈe the L3 per 1000 biting flies; this, horuever rras not possible since neiÈher Garms nor Cheke give data on Ehe total number of flles caught, they dissected parpous flies only), - the parasitic load as mean number of L3 per infective fenale. TABLE 2 is a summary of Table I comparing the data of NorÈhern and Southern Togo âna genin. (One should keep in mind that the Transmission PotentiaL data is not absolute but relative to facilitate comparisons. ) TABLE 3 shows the consulÈants data in comparison to the data collecÈed by VCu at Etre-sanre catching points and during the same t,ime period. In @! epidemiological data is given for vilLages close to Èhe catching sites. Hovever, not all of this mostly pre-control data is reliable and the data for the Comorunity Microfilariae Load (CMFL) are estinates. The table also shows the contribution of each yector species to the transmission and their proporÈion of the total S. damnosun s.1. population. the VCU data on ABR/ATP in 1982/83 is also presented In the text Èhe abbreviations da/si is used for _§r_dqsgo-gggr/S. sirbanum, q for S. uanosum and so/sa for @/S. sanctipauli. DISCUSSION Reliability of VCU data in regard to Èhe torial role of vector species. As all entomological data of VCU has now been stored and is available by STAT at Ouagadougou it, was possible to compare for a given tlne period consultants daÈa with that of VCU. (EnÈonological data stored by STAT should be analysed carefully as some errors uere detected during this analysis. However they were f 3 corrected uith the help of STAT.) The results are glven in Table 3. Comparing consultanÈs data ulth that of VCU lt becane obvious that VCU data ls not reliàble, that ls to say that the species ldentlfication according to Èhe wing tuft colours leads to a rrong inÈerpreÈation of the conÈribuEion to Ehe traàsmission and vectorial role of a vector species at a Siven siEe in Togo and Benin. The urost northern catching point where consultants found so/sa to carry infective larvae (Cheke tggÀ) was Tapounde in Togo. VCU for Èhe same period did nor idenrify 04/05 flies there. Howver, VCU found flies of this wing tuft category to be infective at. Titira which is siÈuated not far from Tapounde. From VCU data it is impossible to assess the vectorial role of e9-. At, sites like landa Pozanda, Bétérou, Mo, Bagan and Fazao sqrs contribution to the transnission is very inporÈant. From the VCU data iÈ can only be concluded thar females belonging to Èhe ning tuft category OLIO? are responsible for Èhe transmission àf ànchocerciasis aE those sites. No differentiation between pale wing Euft g and savanna species can be rnade. During Èhe months May-September, 1983 VCU found some transnission by so/sa at Landa Pozanda, uhile consultants missed that observation, due to a low proportion of so/sa (O.72 and 1.12) in the catches and due to an insufficienE àumber of dissecte«I flies (55 and 51 flies' resPectively). Similar nisleading resulÈ vere observed for Tététou. Although none of the consultants found da/si to contribute to the transmission in 1982, VCU observed an important transtlElon by OI/O2 f1ies. In 1983 both, consultants and VCU found da/si and 01/02' resPectively to be the only vector. As mentioned above the interpretation of VCu data has to be done carefully and all observaÈions have Èo be taken inEo account. For instance, in August 1981 VCU calculated 2,999.8 infective larvae/1000 parous females (L3/TII.P.) of the wing tuft caÈegory 03 at Tététou. However Ehis data is noE significant; although 71805 flies were caught only 4 females 03 vere dissected, 3 of which were found to be parous and only I female uas found to be infective carrying 9 infective larvae. It would be nise to remove from the data base diskettes all those insignificant data. Variation in the species compoli!&E_ All consultants observed seasonal changes in the composition of f1y populatlons at various sites which however were consistenÈ on the basis of the events of previous years. Nevertheless unexpected observaÈions uere nade at Îététou and Djodji. - ltt letétou da/si sas t,he predominant species pair during the dry season and nearly dlsappeared during the rainy season (May 1981: 802 da/si, August 1981: 3Z; Garms eÈ al. f981). In contrast so/sa was rarely found during the dry season buÈ increased gradually during Èhe rainy season comprising 1002 at the end of August 1981 (Garns et, al. 1982). However, this was not observed during the 1983 rainy season when all flies identified were da/si (Cheke 1983, Renz 1983), a resuit vhich uas reproduced in the larval idêntiflcaÈion. (During the dry season prior to this all samples uere so/sa.) I 4 - Djodjl is known for its mixture of conmon until the experlmental ÈreatEent in treaÈnenÈ sq becane the doninant species in both larval samples and adult ldentification (Garms et al. 1982) and it uas not until 1983 that so/sa population began to recoyer and come back to lts original size (Cheke 1983, Renz 1983). Parous and infection rates The PAROUS RATE of the fenales varied siginificanÈly from one site to the other, naidly due to the varylng proportion of old reinvading flies. This explains the high parous rates observed for sq at landa Pozanda and Mô à Mô (between 702 and 1002), uhich is ln conÈrasÈ Èo -the 1or.r parous rate of the same vector at Djodji(152 Renz 1982, 1983). The proporÈion of infected and infective parous flies varied also largely from one site to the other and for the different vectors but does not. show a cLear-cuÈ picture. This is particularly true for the propotion of infectiveflies. In general INFELCIION RATEs uere high for dalsi (Garms & Cheke, in press: 28.82; Renz 1983: 20.12) and so/sa (Garms & Cheke, in press: L9.22; Renz 19832 23.52) regardless of the situation or site. Renz assumes Èhat Èhe high infection rates of so/sa at Djodji and Tététou r.rhich were higher than those observed at Bétérou and Kaboua uere probably due to an increased proporÈion of blood meals on microfilarial positive persons at those sites. The infection raÈes uere lovest for sq (Garms & Cheke, in press: 10.62; Renz 1983: 7 .82). Ttre proportion of INFECTIVE FLIES was highest for dalsi (Garms & Cheke, in press: 9.323 Renz 1983:7.62), followed by so/sa (5.97" and 4.32, respectively) and vas lowest for sq (3.82 and 3.32, respectivelÿ). The mean number of INFECTM LARVAE (L3) per fly nas about similar for both Iga/si (Garms & Cheke, in press: 2.2t Renz 1983: 1.9; all consulÈants data: 1.8) and sq (Garms & Cheke, in press: 2.4; Renz 1983: 1.9; all consulÈants data: 2.0) but. was two to three Eimes higher for so/sa (5.6, 6.6, and 4.6, respectively). :It ls interesting to note that the mean number of L3 per fly tÿas very similar ifor all vectors in the savanna of northern Togo and Benin (ga/"!: 1.6; g: 1.9; 1 so/sa: 1.7) but differed in the souÈhern parts (&lai: 1.6;-911 2.1; so/sa: 5.7). So/sa in northern savanna areas shows the ilsavannatt paÈtern of transmission with a low parasitic load while in the southern parts iE shows the "forestfr type of transmission with a high parasitic load (Tab.2)(Duke 1969, iPhilippon L977). VecEorial role of vector species The vectorial role of each vector species and the inÈra-specific variations of Èhe vectorial efficlency of the sane vector in different bio-climatic zones are still dlfficult to assess due to an insufficient presentation of data in the consultantst reports and due Èo the unknown dlstribution of the parasite sÈrains. Nevertheless, one can draw some conclusion from Èhe data combined in lables l, 21 4 and Figure l. sq and so/sa, vith Èhe latter nore June 1981. For a long tlme after I t ( 5 S. danlosuo/S. sirbanuq is an efficient vector parLicularly in norÈhern parts ofi@contribuÈionÈothetransmissionwasnearJ0Zgy1ng fràn low raÈes ar Bagan to 1002 at Gbassé. At some catching pointsEE-ouLhern parts da/si was often not presenÈ or occurred in weak populations only and tnerefô-e played no or hardly any role in the transmissi_on (Fazao' DJodji, Kaboua) (Fig.i). Ar other plâces-in Èhe south such as Tététou and Alamassou da/si oc".sionally contributed 612 to 1002 to the transmission (Tab.1). Ttre proporÈion of da/si contribuElng to the transnission varied at some places from y"àr to year.afr6t6tou no Èransmisslon by da/si was observed during the rainy season iggZ r*,ife in }day-JuLy 1983 Cheke ( tgBilfound da/si to !g the only vectôr. During the period of invesÈigation however, iEs general contribution t'o the transnission was 8Z onlY. S. squamosum is not regarded as an efficient vector by Renz (1982, 1983) and ffia2)whi1eGarmsandChekebe1ievesqtobeanimportantvect,or/-particularly in the reinvasion areas in the Mô and Keran valleys and at, Beterou.Àt Bagan it contributed 81.52 to the Èransmission although its proPortion on the total fly population was 46.37. onl-y. At Hô à Mô, Landa Pozanda and Bécérou its contribution to the transmission vas 292, 352, and l3Z respecEiveLy' representing 782, 42| , and, L4Z of. Èhe total biting population (Fig.1). In southern areas at Kaboua, Djodji and Tétdtou §g. rras not found to play a rolein the transmission. Horever, at Alamassou lt contributed to a certain exEent(262) to the Èransmission and at Fazao where sq represelted the predominant species (942) it was aLso the most inportant vector (967"). Comparing Èhe daEa of all consultants one has to admit thac sq ls indeed an efficienÈ.veqtor particularly in reinvasion areas. Ttre low infecÈion rates of gg found by all consulÈants is explained by Garms & Cheke (in press) by a low population density of man in the mountainous areas in Togo (the nain source of reinvasion), bÿ a higher degree of zoophily of sg Èhan- of da/si and/or by a reduced susceptibility Eo infecÈion. After the experimental inflction studies by Renz (1982) and Barbiero (1982) one must assume that' there is some evidence for a reduced susceptibility. Honever, that does noE exclude some influence of the tuo other factors Eentioned by Garms and Cheke. The low infection raÈe observed is compensated by a tfigh biting rate in the reinvasion areas which results in a higher transmission*). S. soubrense/§:_sancl[auli is also a dangerous vector of O..volvulus. In soffiern parts omgo antBenin more than 9OZ oL the transmissi-on was due Èo ;;l;;; â*[àpi-"t Alaiàssou (this, however may be due to the spa{e data for this siÈe). In northern parÈs its contribution to the transmission was lower than in the south but uas sÈill remarkable. As iar north as Tapounde and TiÈira it represented 342 and 2lZ of the EoÈal biting fly population and it contributed 152 and 212, respectively Eo Ehe Eransmission (Tab.1 ). Ttre significance of the disease transmission ln regard Èo the differences populations or strains in Èhe various vecÈor species still s is also true for the EPIDEIIIOTOGICAL SITUATION ln Togo and beÈueen remains Benin. 0 volvulus unclear. Gebassé seems to be the only place with a clearly identified *) It seems that in the Mô valley, at Landa Pozanda and Bêtêrou Èhe role of ggis similar t.o that aE Guêna, Burkina Faso (Philippon 1977). -6 ttsayannatt-onchocerciasis transoitÈed by da/si. It fits uell in Dr Renmets regression llne of the correlation of the prevalence of blindness and the CMFL in savanna areas (Fig.2). However, there are also other places nhlch uould fit into the picÈure of ttsavannatt-onchocerciasis but nhlch in fact after Dr Dadzie are places of a nixture of two or tuore Onchocerca populations. Bagan is such a place of mixqd onchocerciasis mainfy transnitted by sq (822) buÈ also by da/si (18U), there the CMFL afÈer Dr Dadzie shows an increaEfng trend whlle Èhe CMFLAC is declining. AÈ Hô à ltô atttrough .dalsi is the nain vector (712) beside q (292) theprevalence of blindness is rather low (0.92) conpared to the CMFL (33). lhis is also t.rue for Bétérou. AÈ Bétérou and also at Alanassou all Èhree specles/species pairs are lnvolved in the Eransnission, da/si belng the most importanÈ yecÈor. Titira and Landa Pozanda are also places of mixed onchocerciasls transmitted by da/si and so/sa, and da/si and q§, respecEively. At both Tittra and l,anda Pozanda one would expect a higher prevalence of blindness considering the high CMFL of 80 and 50, respectively (Tab.4, Fig.2). After Dr Dadzie rforestr'-onchcerciasis is only found at Djodji and Kaboua where so/sa is the main vector. In the south at, Tététou rrere a CMFL of 9.2 and a prevalence of blindness of 3.72 was observed so/sa conÈributed most to the Èransmission (922, beside ga/si contribuEing 82) carrying 4 rimes more infective larvae than da/s__i. One can assume that these diferênces are due Èo the physiology of the vecÈor species and noÈ to differenÈ strains of Ehe parasite. Unfortunately, nost of the epidemiological precontrol data is not reliable and only very fev data on G.IFLAC is available. Furthermore sufficient lnformation is lacking on the nature of different populations/strains of O. volvulus and their associaÈion with the different vector species. Above thaÈ Èhe degree of zoophily/anÈhropophily of the vectors and the role of 0nchocerca of animal origin remains unclear. Therefore at presenÈ the relation between population/strain of the parasite and the nunber of infective larvae transmitÈed by the vectors and the establishment of infections of varying severity in the hunan host can not yet be explained. Reinvaslon The main conctusions one can draw from the consulÈants reporÈs including those by Baldry (1984) and I'lalsh (1984) were t.hat: - There are probably many sources of reinvasion for the various regions in Togo and Benin. - Some evidence vas furnished to indicate Ehat flies reinvading norÈhern Benin (da/si) could not have been coming from the east (Nigeria) but from Mono(small number of early flies) and particularly from the Ouémé and its tributaries, rhe Okpara and Térou (most flies, nainly in August). - Reinvasion is certainlÿ the result of long-distance movements of flies assoclated uith the prevailing south and south-westerly vinds. - In Togo in Èhe Mô and Keran valleys reinvasion is mainly from the Mono and Anid river systetss and from Èhe nontainous areas LocaÈed south and south-west of Mô and landa Pozanda as far as 200 ko (R. Asukawkau). - Reinvading flles can arrive from more Èhan one direction due to changes o7 of surface vinds from south-east Eo souÈh-uest durlng the Donths fron May to SepÈenber. CONCLUSION The studies carried out by the consultants showed that all Yector species da/si, sq, and so/sa contribute considerably to the Èransolssion of 0. volvulus in Togo and Benln dependlng on the season and area. S. is a very efficient vector in northern Togo and Benin but It also contribuÈes considerably Eo the Lransmission in the souEh if presenÈ. S. squamosum and S. soubrense/§_._ganc@gli--are also efficient vectors in central-T6!6aîil in souttrern arèas, respectively. However it remains unclear as to uhich extent the two latter species Broups cont.ribute to the transmission of the seyere savanna forn of onchocerciasis. The nature of transmission is further complicated by variations beÈween years and seasons in the vector species and age composit.ion of the vector populations. VCU data based on the vector identification by the colour of the wing tufts only is not suitable for the evaluation of the vecÈorial role and the transmission potential of each vector species in logo and Benin. Consequently false species identification and mis-inÈerpretaÈion of data occur caused by mixed populations of pale, nixed and dark wing tuft flies within so/sa and gg in Ehis area (after Reni (19S2) 822 o1. sq females and 167" o1. so/sa Eelong to the Ol/O2 wing tuft category). Howeverr-a reliable species identificacion is crucial for the analÿsis and inÈerpretaËion of the data collected by VCll and for the correlation of EPI and VCU daÈa, even if the 0. volvulus population cannoE be identified yet. RECOTIMENDATIONS It is therfore recommended: - to select in Togo and Benin a couple of key çatching-points t,ogeEher with EPI and STAT (porbably Titira, Landa Pozanda, Bétérou, Mô, Bagan' Djodii' Tdtdtou, and perhaps Fazao). - to identify correctly Èhe vector species and their !tsRs and l'lTPs at those sites during the whole year (so far, correct species identification by consultants uas carried out during the period of reinvasion only). - species differentiation should be made by the identification of the ratio thorax/antenna Length and by the form of the antenna segments. Material for iso-enzyme and cuticula-hydrocarbon analyses shouLd be collected as welL as infective larvae of 0. volvulus for DNA probes. EvenLually sinple lso-enzyme analysis could be employed at a sector or sub-secÈor to separaÈe sq from d'a/si and gg@ lf necessary. Ttrere are seyeral possibillties to carry out such a study: 1. VCU collected and alkohol preserved parous flies could be identified and dissected by neàns of a 1§A (Drs Cheke, Garms or by the nriter). The materiaL tI should sEill be available at lana Kara. Ttre results should be presented on VCUifichest, rather than ln a report to allow the storage and analysis of data by STAT/VCU. 2. A reltable VCU technlcal assistant could be trained in the identlficaÈion of the vector species (by Mr trlilson or by Dr Garns) and if necessary in iso-enzyme analysJ.s (by Dr Meredlth or Mr Boakye). 3. It vould also be possible to have the study being carried out,_by Dr M. Denke of the r'Institut Nationale drHygiene" at Lone on a cons$ltancy. Dr Denke would also need a Eraining ln species ldentificaEion and iso-enzyne techniques. Besides Dr Denke ,outd be able to identify in the flies animal filariae which would give some evidence of che exÈent of zoophilic behaviour of Èhe vector specieà. This sÈudy could also include f1y catches on some animal -baited traPs. Furthermore it is recomnended to study the species distribution of larval populations, parÈicularly during Èhe dry season (preferably from February toÀpiif). The rànaining breeding sites should be napped in view of future Iàrviciding canpaigné during [tris season. There is founded hope t'hat so/sg popularlonà couid be substaàtiatty reduced vlÈh long lasting effect by only few treatmenÈs. ACKNOI.ILEDGEHENI§ I would like to express ny Ehanks to Drs Dadzie, Remme and Mr Zerbo for very useful discussion. I an parEicularly thankful to all members of STAI, Dr Remne' Messers. Dupuy, NrGadjaga and Soumbey for Èheir paEience' help and support. Furthermore I nish to-eipress ny thanks to Drs Samba, Philippon and Subra for their supporE. I Kar, dat 6,1 sq q o KANDI NATI TIN GOUo :\ :) 5O Km Fig.l Locatron ol catchrng sites LAMA - Ma ; Mô gridge Landa-Pozanda Bagan Kérémou Alrbori Zougou Gbassé Landa- Mono 8étérou Fazao Kaboua Oiodii Tététou Goubi Alamassou M'bétékoukou Kerou Oti -trtiounga ,Nabou lgou Tirifa Tapaoundê Tipaoti Pouda Tchelenga QfP \1rr6{ar., @ @ @ @ @ o @ @ o o @ o @ @ o @ @ @ o o @ @ @ @ o a .l ,{r a ',1 , \ I\) , ,a(), It FIGURE I a üONI'RIBUTION ÏC TRâNS!'IISSION ti ËÉ îl ÊlÉD(9Hh zz ! o^ Ex =J! o L o{. JItz -33-()fr @ \oo ol- oN ,F oo ,F o|o oa oN a o t,l o 0- lrl 1' o o+. o! L o rF a =, o oÉ, (rf) V) E § à \ § a a ts § q § § § § \ Eï È t-) § §aU o @ tr o|@Fr..OO'trONoN §§\ a I (r §\d{ I I tr: r§ tr'§ tq\I h ù § * t,\t _§ x\r x tr xt d x t È I .ï\ t \' \ x§ Ix x ÿ§ .\§ NI x .l x x x o t ü II x XX x x o o sseuPullq lo ecuelD^eJd'PuDls tN ! oÿl N qt\@lfl oo<11@Nr É6lft@NÉNN €- t/to?4l,t-(Drftul$rôÉlflMo{ t t4!t @ oN n (.1 o t1 N!t o o !1 t\ n a4 oNo§ontlllüI ll u lt ll lt lt llEll UJ II l![ I <EIÈll .( Illl- ll TI NI II!I ltz1IIIIINI lt fl lt u ll llI Ell uJ llla ll llû[ .IIIFI IIl TtIII!II!III! 0I T üIIIII llI TI uI llI JCt <r ul l-EOJFl! oêoo oo?3o oeoo oGoc. oooo r\ôoo NdîN @<qd mUl{t@noa\ Nooo NNTlO oo oo oo oo oo oo tÎo <o ÿro aO oooo <-oÔ ôooo €8 11i100 <rtâ.30 al/, à-OO rJoo oêo ô o.f 000 ooo o 4..3 ooo ooa o o.3 ooo ooo ao E 1rlù },i) (! l- Ê û Jf û- l-()2 <t o o lr,(Il z trt tl(üI o 1, o u IJ IL !âJ ulrlÈ aJ) J o. oeoo o].r@o oÇNN oo rioéo o-on oo oo r oo oô ôo oo I00 oeooo orl .c00 ({€ r oo ({d oo @ oN oo r'l o aO o NON O6(Dd@ c.t<n OfrNO ü)r1({o({14l.O Éd@l? rJt ocoo <G<ô <o11 êtiN t'rOF.O <OaO <ç I oo{}1 o o !ô o o @ N o o ç looE)oo .J, oa\ !tN o rtÔ{o oomoçÊ(l o o(} I oo N O]lONtt cù)M(DOq'rlam@ üt N ÈN o D o n o I o{ o({ooNNON',t nl\rîo <t€t\Ot§€N oooo ôNnO not nN oec{o !1 @ I oo -< di €Ç I Ô{Nd Nl.!1 r ul r 100 <rN oON(l oo oo z N À t- o- o !1oo -itN ?1 0aotô { on I o{ o66 F. tft { oo.? oooooo ooo À€1 utE CL I N z it €11 r)N aô rtot no oo oo <({<1") À-oNü)dl/rO !tÈiâo <odtoF{rto ooÇo I N: lv) o rfl{ - '8..{to -@ôrt \l{G N6 -N Na\ .t4t1 { .ôa@@ €- -E t.Ià8 z 'tC+, I I \ <ar\ù{t\G (9 O< E ",r,.,, É.,. n t4 o @ -Noi^f,\ @OOI)\rlG<@ô@lJt\t\FOÀt\t\I! rÊaEoGo ft \@ oô ùlôt\ @ ul (0 11 |') o @@(jlr\\ < 0r€€ dfioou(J t-2 oÀ oz f ot- <t(J N14O@{ {t'{O NnÇN{7rÈ)t { h.tiOrtooo ooo oooooo 0- o $N tt CD o tl lrt{ l:l<N-oNn€@€Élrl @o{ o<toÊ'rZt\cl Nl\. lflN(Dct.@\tâÿ,G-\({(D(0cor\\ ôd I .â I I2i6<Ültr(t JOGO(Jq GD 6ooç <@ coE-HlFO FCI t @ ldÇo@z\fo o< <ll&o FCI E :l2 Irtr o o =ah clzE <lê UI I I I I I I I I I I I I I I I I I I I I I I I t t I I I !t I ,: a ll[{[.üq ll ll ltI itIl! çIt{tl <t llI ll ll ll llIN[.It ct[€ ll ll ll llI rl I ll llI ll ll uIIe{ ll llII fl ll llIilt1[{fl:( ll !l t!( I; N!l .r,{ lld rl (\t frlFt cÉl Ê. rl ;r t' l!I llIir oll. l! < il rl ll ilI rlIt1iln(< II ll flIIIt€ll .I.3INI It llII lt llI ll llI lt llI ll ll ll il !t rl ll llIil J i! h:l crl l- € tr, €({ tî n zzü IJJlr, ÈI:r-ofaooOF o À rfl t\HllJII Elü ld <! ll l!üllxlltl F ll trJOllrL zil< [ ]1 ÀollJ. .r IOü l-\lllrl llmIztlI rI ll .No[ùfllollFoll .I ollI T III ll ü lt lt ll TI! ll ItI lt üIIIlI TI üI ü lt ltIII llI llIIIIIII üIII uI nIII ll ll llI ll h l! ll l: !l( h tl ll :t .l II lr lt llIII lt ll lt llI T ll flIII llIII ltI lll ll llII r. F. Q { oqt N 6 Fr N lr') 0-N l- q 14 rf) N 1{ E, UJ û. It', {L r È. c{ { o € , ü IJJlr, üIrtsoüooo E lr,lr 6 f0 c{ lro o lIz oIu, ollEllU <tüflllr cjXlÀ (,lllui .ü ut llJc È3È(J ; llI ti llI llI ilIl ltII llIII ll üI! E lr, IJ. it ù, 0. mÀJ.IF' Ef z <t r!g. a o Efa o z L <r o o N 0- t- o a- UJ 'I{ i l..t Y ? oô ooo «mo Nt\/, t\{!1|âa@qtO@aÉN|0lrlt N11 oo0 ooo <G5t C,ld« €< !aOMlr)< @ ù'l u'! tr'l !'t<Oti @{G,«(0O.3 lr'l oooo -Où?(tî N<Ç 100oa4 <to€( oorl((t< ôt <t t1 §t t\<ot'O<1,)<{!r I dot\- clN!Ô- N C3|1 I &r@{€ <tt.!c t{ o EUJ.lrU! .o l-HoâF Ntî]1 (f tf o G Nô tl 1"1 o @a\O onrt(D60@@60(0û.O(,.6O ({ 6 @iN @ltl\ oooll\(GÉ@O.qlrrt\t\F(DÀNF tdaqcuÉc @o € o <(Do\zr\ <lt NN!'tNan cl @\MMt a\N(D(D aOÉ\\ ê+, I O I IZ t@rûlrr J(oqqoü 6o{st -@ <oîta HlFO F(J rto @ trl <fo@z\foCtÉct0. û. F(J @ 0o. (l6t\ t0 Ir, a ù/, toa @oO+r\\ <c{o É0claoo OOOdist!t <- 1..]ô ô,L :ct-tr\ oll\J <t 0- q. It- t4 t4 IL .J, IL IF Nrl .3J où o oÇnOC{!/rNiÉÉoo< (1 O. (.1 O tO <r N OOOOO a a1 tlt ôo I o.f s o I NN(,.ooô ooo N lli .f <r - OO.â t\ O ooo t4 0t1 c{ ooo o ooo.r.n{ I o1? ooo r oo o o ooooo ooo <tt- a lu €NÔ,Nc0i@N(D@ ooo <{Noljl oo I oo o{ tUJ. LL(n .o FHoaF eooo ôrâ@( !1 0ôo t] nç( OOONs <trrll ql.3lflrù'ttî tt {( tl ç<( aG@( o o 0- OIOF orlI,J t- th 0. aF o]4OJ û. an À Il- Ùl ]1\J oE. À an il ilIIII ilII ilIIIIII fi( il ltIIIIII llI ltIIIIIII ll ltII ooo oôo NmOOI€ ûrfANÉ6ljl€(DS< ooo o., oooo t- <ra ot-z ,-Jf U!zoù ôo IrlN {({ o M € Fz o 0- oz I(J t- (J IIII ll ltI I l-t al'a al f^ rtIa ,,).,r' ltIII ll( ll ll ll ll ll llII il lt llIIII n ItIII ll lt llI lllIII rlIIIII ll llIIII ll ll tl llI ltIIIII lt ltIII ltIII lt lt llI llIIII ltI lt ltII flIII lt llIII üI ll ll llIII lt llI ltIIIIt üI .:..:.ù. (t o r'i N ot (.r r.'!.3 c.lNt.ü€(l14ù<r{ù'l .fd C.l Cl t'' lrl ül O < tl { F. tl €æo(.rstr\oçrlulnoo C.l C I d (.1 c{ }1 11 C.l Cl .{l€tlF\O{(DOt (IlOlî.Nr.OêOmn Ol<<G.-oo N N{ o@N(t' <- Oa< O,3<tNln(D{+U'l-i11<t .Nçô .<1<t0(t < .:.O!lr.OO(r€nC{}1O ,3.fNooNn . §F)dd lf) d ao.Snmo!){€€{ol4<N .0' C,l d qt OO<r.'UINOOOr\€Ocr . oD+n € ô.3<Ot'1001?$emNel i SôaG hÇOO:@ .{ o It', t\ 14 N t'l û. O <t Ol Ot trl N e.l{qn.3ON-O<<P} a<ct14.{)lrlioêo.)N €F.O(D.{rNNnnNOO .J' ti E 'nIl:C(r oÈ fOtÀ>F- N' oz\oo t!F J :lû-x EOn '.L tJ- o rIÛN - ,i Üro\(aû rrtl t 0ltIIIII üIIII f,I 0I ü !l OC.Onqd .fON{NN zOlla6o\ooHtlç a z <t tZ r- (fFU c F 'iü N.-t'- oz\ooUE â z J ,trr Nj UJ uj. 7- ii Hll zl r.! !lùll U\ll<t o it:À oIEOilF- ll it <t ll all <llJOlllrIrEI [U il xllùlliJ ll lltt ltltl i? trl ({ û O ln tî !1 N (I. <t O N(.l.olf!c.tM .<? .!'t .? oa a ? a Nf @a G Ël N t4 <tÈ 'a(f OZOê i€o€M<t<O <+ Ot4N<Nilstu)OG-fm!1t1 0 t4rt].10cl(!rLIO<< OflJruJ( OE <tHOOOO<t<tü ZOI'1<tt-OOHQIJ<<fCOâElrlc<Fzt- oNlIo<lF dü dlt x <f UI O <l qt <t D J lrtoor-JÈEtrrL:4o<lt- tdo JJ I t-!! ,r' .1a.,J'-,1.. ..1 ' ' Jr \r ': r'-: +7r i* Y II llI ilIIII! $ ü üI lt flI llIII itIII lt üI ü lt I t i I