Document 2jODy0X0Rpn2xxvkxN1Zoo75N

v polychlorinated biphenyls in wild birds IN BRITAIN AND THEIR AVIAN TOXICITY Ian 1`nisn, D J. Jmoaih A V W. Mooai Msnkt H'ihhI Experimental Station, Ahhvts Ripton, Huntingdon, England I I 'ABSTRACT flu- amount* ofpolychlorinated biphenyl (PCB) compounds present in (he livers and eggs of wild birds in Britain were determined by gas-liquid chromatography. Most of the $59 specimens came from predatory birds obtained between April 1966 and August 1968. PCB was found in terrestrial species from most regions of Britain, in all the individual and bulked samples of seubird eggs examinedfrom one west coast and two east roast colonies and in most of the freshwater specie* collectedfrom thy mbHunds, east and south of England. The highest liver residues were found In fresh water ftsli-fx'cdinj birds up to ca 900 ppm/ and bird-feeding raptors fup to 70 ppm . The levels present are similar to those ofpp-DDE. An indication of the avian taxied} of one PCB was obtained by feeding Arochlor 1254 to Bengalese finches. The estimated dose rate for 50 mortality at 56 days %vas 254 nig kg day. At this tfose rate the calculated mean liver content m 345 ppm. It has only M3 the toxicity of DOT. but could be more toxic at low doses because it appears to ha\e a more itratlual mortality curve than DD T All birds dying from PCB had enlarged kidneys amt before death same displayed apparent leg paralysis or boslv and wing trembling. It was eonchtded that PC B is unlikely to have caused widespread lethal toxitv in wild predatory birds In Britain, but could he a component cause of the present breeding failure reported In severot species. INTRODICTION Studies of orgnnochlorinc insecticide residues in eggs of sea biros and in livers and eggs of predatory birds have been in progress at this experimental station since the early 1960s (Moore & Tatton. 1965; Preset. 1966). The chemical analyses were made by gas-liquid chromatography, using electron capture detection. In addition __________ 3 fa**** Mm. (I) (ItTOl m. S3*. Pi*SIhki CMnpafty III, Kn|lU-Pnritl in Gfmi BriUta *\ HONS 086963 4 IA ' PAl.m. I). J. JEFFERIES. N. W. MOOftf to peaks corresponds to such organochlorine pesticides as BHC. dicldrin and DOT and its metabolite*, it vs as not unusual for an additional series of peaks to appear on the chromatograms. There were as many as 10 of these peaks, corre-. sponding to unidentified compounds. Roburn (1965) established that these were organochlorine in nature and for some time it was generally supposed that they were either further breakdown or condensation products of the organochlorine pesticides or possibly loose compounds of these with natural products such as protein matter. In 1966. however, Jenson succeeded in identifying a series of such peaks, in the tissues of 2t)0 pike (Esox Indus) from different parts of Sweden and in the feathers of Swedish white-tailed eagle (Hafiaeetus a/hidlfa), as corresponding to polychlorinated biphenyl (PCB) compounds. Following this Holmesero/.(|96?) showed that the long retention time compounds occurring in British wild bird specimens were also PCBs and since 1966 these have been quantified in addition to the organochlorine insecticide residues. This paper gives details of the amounts of PCB that have been found to date in wild birds and their eggs in Britain and also the results from a laboratory experiment in which a PCH was fed to the Bengalese finch (Londutra ttriaia). ANALYTICAL PIUXCDL'XE The material obtained from the laboratory specimens was analysed at Monks Wood Experimental Station by M. C. Frehch. First, all specimens were ground with sand and anhydrous sodium sulphate and extracted with hot n-hexane and acetone. The extract was denned up by the technique ofde Faubert Maunder et al. (1964) and subjected u> alumina column chromatography. The final hexane extracts were analysed using gas-liquid chromatography with electron capture detection. The columns used were packed with Silicone/Epikote on Diatomite CQ. The Arochlor 1254 fed to the laboratory birds gave a series of six main peaks on the chromutogram. As there were no changes in the relative peak heights compared with the standards (M. C. French, in preparation), quantification of the residue was simplified by comparing the height of peak 2 with that of the standard. Confirmation of the quantity was obtained by the second technique of comparing the total area under the peaks. Wild bird specimens were analysed on our behalf by the Laboratory of the Government Chemist. The technique used was similar to the above but quantifica tion of the PCB residue was made more difficult owing to the presence of organo chlorine insecticides These were separated from the PCB-type compounds by silica gel column chromatography prior to gas-liquid chromatography. Other methods of separation are discussed by Holmes et at. (1967) of the Laboratory of the Government Chemist. The lack ofcorrespondence between relative peak heights on the chromatograms of the specimens and of commercial PCB preparations, HONS 0B6964 KX YOU OKINATH) eiPlltSYI.S IN Wu.iv IMKIK in ...... nil in some invianccv ihc complete absence of certain peaks from (he former, prevail* (he determination or ihc amount* prccm b> direct comparison. The assessment of (he amount* of PCM was therefore based on peak heights produced hy the electron capture detector related to standards of pp'-DOE and dieltlrin (nr slmllnr technique used by Rischrnugh </ al I IMS)). As the pattern of PC M peaks is frequently similar for dilferent samples, and as the same method of com putation is applied to all. the values reported reflect the relationship of tne levels between samples rather than the absolute level in etch. The figures shown for wildlife samples should therefore be interpreted in this light. Results up to 10 parts per million have been given to the nearest, between 10 and 30 ppm to the nearest 5 ppm. above 30 ppm to the nearest 10 ppm. and above 200 to the nearest 100 ppm. ST CIVICS OF WILD BIROS Specimen* examined Analyses for PCB have been completed fora total of 196 livers from 33 sr ccics and of 363 eggs from 28 species (see Table I). The specimens were obtained between April 1066 and August 1968. The seabird eggs were collected from So-It Head. Norfolk: Si Abbs Head. Berwickshire: the Mull of Gallo&ay. Wigl.>unvhire. and one of a great skua (Siereorarlus skua) from Handa, Sutherland. 1 hey were part of the annual sample of seabird eggs collected for monitoring ir.*ccticides (Moore R Taiton. 196$: Moore, unpublished). Most of the heron (Arden clnereaI nd moorhen (Gallbnila rhloropus) specimens came from eastern England, being part of the material collected during a detailed study of the heron (Presir. in press). The bird of prey specimens were sent throughout the year from various parts of Britain and represent part of the routine continuous collection being curried out to follow changes in the insecticide residues in British predatory bird* (Prestt. 1967). Most of the eggs were collected w hen fresh and after receipt at the laboratory the entire contents of each egg were blown into a glass phial and stored at - 20 C until analysed. The liters were removed during autopsy and also stored in glass phials at -20'C until analysed. Geographical distribution of PCB Terrestrial --tubs can only be used as indicators of local conditions if it is established that they are resident in a particular area. A total of 138 liver specimens from terrestrial species (f.r. birds occupy ing terrestrial habitats and feeding mainly on terrestrial organisms) has been analysed to date for PCB. All but eighi were from birds of prey: 75 (54 %) of the specimens were from largely sedentary speciesgolden eagle (Aqulla ekrysotlos), sparrow-hawk (Aeclplter nisus), peregrine <Fako IAN PRtSlT, D. J. JEFFERIES, N. W. MOOftli table i *r ttsuAs m intn a> Ka ui w*j> nun k sarrArs <1966 *1 Bpdkipidae Great Cnu] Grebe Fudkrpa crarmrm Slavonian Grebe A. tmrkia UtUc Grebe K nfiwth Fhalacroeoracidae A6Mn Shat Fka/ocrocvrax arist<Mriti Heron Ardea enterra Heron nestfey Bhlern Anmtu sttffons .Whooper Swan Crgmr* nyw Fakxwidac Balearic*!* Haematopodidae Golden Eagle Aqrnla ehry^ftvs Buzzard Buttomutm RooMet*ed Buzzard R. taxpu* Sparrow-Hawk Actifrirer ms*3 Kite Bibas mdrms Hen-Hanier Cwatx cywma Osprey Fmndiom bu/iaetm Hobby ftlrr mhhmfrv Berctrinc F. ptrtgrima Merlin F. emitmAmnmx Kesriel F a!u\ Water Ral ibAu <tqmnktu Moorhen Gsftknda chforuptts Oystercatcher Honmhfms ustndrgmx 4 2 2 |( 14 2 2 J |g J |( j 1 2 | 2 47 | 30 40 *I1 4 1$ Oca900 1-30 I to 0-IS OJ 1-40 0-2 3 5 20 0 0-90 0 25 9* 5 4 05 II rot 20 5 o O- I 0-10 0 0-2 02 1 06 2-4 0-2 015 Ol 2-4 0-3 *0NS 86966 (NH.YCIILOftINATED BIPHENYLS IN WILD BIBOS IN IIBITBIN CWliWM* Seetopacidv lywim Hwelfci min Rjffd* PWwf ChvAan AMdSr WcgdroA Srwitfmj rmrindt Lnbc Iknini GR {#bxmmr Wldhufcj Cn i. rtoftwndu Uttwakc Am nefciTjfa Common Tern Swam h>/.i Sandwich Ton S. mMny> f0 19 Akkiac RazorVTT ^f/c^ ft*nA/ Little Auk Wwaudlr Coitleniot C/rar ewfrwr II 14 Strigidae KanvOwl TjimaBm Little Owl Athem nirfm Tawny Owl Stri**fpn> Long-eared Owl Asia nrt Short-eared Owl A. /towwmm 22 050 90 3 07 I0 3 02 Atcedmidac Kingfisher Atmtr* attfri\ 14 0 4Q KirwWtniitx SwaRow ff/nwii unrnv 1t Corvidae Laniidae Raven Carm mu 20 Carrion (Vw (*. rww*- I0 Rook f. /hy>Hy.i I3 ftGreat Grey Shrike L*mm exmhit*# I *Stunwdae Starling Sturnus rmfgerh I a n tO 5 I I0 I 25 I( 13 5* 14 1-2 t-0 40 1 10 2-4 II 7* 16 5* 4 O-S 4 04 I0 Total 196 363 *Cy$ 6967 Reversed liquid-liquid Partition in Determination of Polychlorinated Biphenyl (PGB) and Chlorinated Pesticides in Water IMP AMU*' l>4 Steen Jeerin' Chr-ir-v. lVnrk.-y c/J-mJ.V/.-. 5-f0f Of.dsK*.Sit-M l nw metheO bawd an an application ol the Hmitm urtrttd llpuld-llouM partition melhed It doacrlbod Ur Ik* extraction at chlarlnatwd pntUd4a lrm wtltr. u Mil mothnd Uit wtltr It pM*4 thrnu|h t filter a arame) eenlelnlnp t mixture ar e-undecan* end (ireewax MM mtirattetrate an Chromoterb w, and abttrbcd peetleldei trt eluted wifi) petroleum I'.ktr (M ml). When detected I) moan* tl t |tt rremitofraphle mltm with in tltdrtn upturn nutter, ten atneltlvlty lull tm/m' tl IlMana with t Rohm and Heat (Si and also found to be rf.s-.rr: ,'vr lb* extraction of chlorinated peuiddn Aram tttcr. Ar- extrecdon at tht Altar material (art, however, poor ractvtry. Oood roaolta both In (itraclioa and recovery from Alter material ware found la a method bttod an the revontd Uquld- #4id partlliotl method olth partition Horn water to hydro- phobifed earner totted wish a 'Jfcjhi'J: miSoea> phut Mowed bp recovery from the column b> meant of a itr- amount ofpetroleum ether <10 mil. Ownto uertwr riAU, many atlempta have been made to mmihttNTAL Memlnt tht pretence of cMorlaalad kydrocarbom la otter. Reaacata aad Eedennt. n-Undecanc. (purum. b.p. 195- Ik two methodt moat often ueed are bated dtker 00 eoo- IM *q ie ehaken wfjl| portione of concentrated aulfurie arid jmout telrtction with tn orituilc aolveat (/, 2) or tdeorptioo a a flhir conuiniat activated oarboo (J). Heart methode a, however, rather time eoanmiat owlnt to a loat ex action lime, both from the water and from the carbon, the ecMklvky at the dWbraot methode deponde meetly pa It way of iktoctkm and the mod volume of water. Thm (Aa and Weyman (/) extracted eavertl hundred Ntcre of dkr with petroleum ether. Hie epead pf extract|o wat J-1 KterAour and rvoovary ofadded paailclde wee 12-lflOJ^ (px) patll the arid h eolorleta. The undeeme it tbm patied throuih a column of eritmted tlundne (12 boon at 230 *C). Petroleum ether (Skellyeolvt I, bp. <0-70 *C) and acetone CM-) are dMDad Ihfoiwh a I-meter column AIM with aadeh and laaulated with alma-erool end atumiiwm foil, lha head pf the column h Ailed M a Mil-heed eondemer adjusted for KX rettux. The eolecnla are lotted on temper cbroeiiete maph pfler evaporation te the eamc dune m hi the analytic. Cbrhdtmi monoataarale (OP 27) waeJhlaJmd from AmtMfcri fiAphtbprlAC Uhotutarica Inc. (Sremoaorb W Iname of compounde waa detected with an dectron affinity ewetor at the 0,1-MO ppb level. Raeoa and Mlddkttm (J) me MHM Aeeeb. HMOd traated, end erid waahed (JohatMnnvfllpV Aulfwfc arid rnt pa. (MarckX at activated earhoa tad 3000 Ikert of water tad extracted k chlorinated hydrocarbon from do carbon with (Mo dem. The rooovery waa 7S-MX and fin LS-ppm tend a readied wkh infrared detection. Aeetyenbach (I) Aeuad UII houta* extraction with ehlorofbrm waa aecaaaaiy tar ptetc recovery at the chlorinated hydcocarbone Item lha atea after prevloucly Cbyiaa the (her material tar two The Wdr cohwm wm 20<m X I em l.d. wkh a tlaaa-Aher dkk O I (IOO-l2Aa) A Idnd madaated caatrifuaaMon tuba waa mad. The am ehromatewaphlc column area koraallicate ahua, IM cm X AJO ten. All the Afcecwafc la waahed with deterpem, heated la a mixture ofeulfurlc arid altrlc arid (4:1) te T and rimed wkh dhllAad water. After dmidirn, Ike Atemwire la checked by riwkiac wkh n-heaene fiwm whn 10 pi ie Infected on the htM*C. aec dwomaWAfepk. The pae cMmwaieemph wkh EC- lecent pcoareee of there two methade aH bo found la an datacuc wm a Variaw Aarcjpaph 204, with a Spacdemaa xdem review byThornbar* and Beehmaa Mi 0-1 mV racerdar. Paper meed wm 12 techarihsur. tar Intention In the prceent work haa been to develop an Oaa Cbriwutaarapky. The cohitwm are HMOS-tnatad ear method than the above mentioned, hi vriOA it oouldte^. Me to reach the peapemd level ef etdorinated hydrd- etene in natural water In Sweden. Wfluiff of tht ohopMlie thtnrtif at tht chloripMtd . and AIM wkh K-.|A0 atarii HMDttneM Chramoaorb W aavaewf wkh ritem 4* mrikyl rikmap tk CN> Aft or IJ5 AMOttWaoea oil <0P n. Tboaarrior im ia akrofew pvriAod Wkh a Ateeh mofeeutef affier*. ON epead la ahowl 30 ml wo nk' utr e and Manor temperature ia JOS and MfcMct* t potiMt mUM ht btttd or tht oonv *Riy titB ftwttB phntpovtitiOR MtthoBe 2te30rivTe,DreDt:fTnSkkmThteimoeoloamf 2a0 egairmnpdtmrat(uerabwoaaucthiwoacan (Milieu at ariaale malarial from water by meant of Preparelire at ephnaa Material. Tea iranaof Carbowax marine XAD (a treat Halted polymer) wat Ant atad by .I NWi ^MRidMiiimI PIffcr VTnNdAH^ ra'f^ll tU^hniWla ilva^lnwJimmlmba Krletiaar via 4TD, 114 AkiHeha TUeemi addrew, Uatteveihieilalla, Kami L Uhnaa. TteOT, hi m hi nmn hi ^ thRllMr Btsk. OwwOaMtI hOOHa^mhowlt^tie mpmnte tfCWkriMavMMHaMomdw W^w k kOTMWII ffiVtOilOtVii^s^fitll hvPvp addMipel aeatoar te caver at aupyort The acetone la evaporated ia a rotevepor apparatae under iliehtly dlmln- L Kxhe end C K. Weyenan, Arw> Cimi., 04, IHQQCCri. A A IteHh aad & M. MMrialaa, AM, M, I12P (IPteh A. W. OrkOmbarli e(W.,"TW 14mlIlk.Own r>4 tlwlurmml ef CMoriaated HyAoearbon Netieidw la Surteee Welere,'' U.A. Depenmeatefthelmerlac Pedwal Water Mhalan Cbairel AMnieuaiioa, Havamter 1044. * 4W. ThetWhurp tad H. Oethiaaa, Amt. Oar, Cl, lk ONt). the catenae material ere waipkad law tea column which Ie thM Biitd it tht oudfetf at Bit wtttf ctattiiiiv by mm of an all-clam Otilno. b laboraieey work, the tower and of the column la oomiacted to a water auction pump and In the laid work, to any other kind of vacuum punte. The vacuum (N fiahm A Hart Oteat * Mnm, 44, M (tea. IT, INCX HUNS 0B696B MUIYTICM. CHtMIOTRY, VOL 44, NO, II, NOVEMOU 10T0 144* t BOLYCMLOBINATEO BIPHENYLS IN WILD BIBOS IN HkIIAin MUNS 086969 MONS 086970 I 1 PIM Vnil.DKINATm HIPIU NYI.S IS WILD ItiftOSIN IIRMAIS |! will) more than 10 ppm PCB present in (he liver have been obtained from all parts of the area represented by the sample and only 12 (27*',,) of the 45 specimens had less than I ppm. Again the egg analyses give a similar result to those from the livers, with < I ppm only being found in 38 07 \,) out of 102 eggs analysed. These findings show that IH'H'has been obtained by freshwater lish-fccdmp species in all parts of the midlands, east and south-east Tngland. and suggest that about t, thirds of (he individuals m this urea may contain PC B in considerable amounts. Murwif.- The Scott Head egg sample contained 10 sandwich tern tSlvnni antf*icensh) eggs analysed individually, and 10 common tern (5. lurumh) egg> bulked: the Mull of Calloway sample contained 7 bulked guillemot (( na aa/yan eggs. 6 bulked shag iPhalacroeorax arlstoteUs) eggs. 6 bulked razorbill ( Ucu ior<(a\ eggs and 6 bulked kittiwakc (Kissa trufactyla) eggs: and the St. Abbs Head sample *5 bulked shag eggs. 8 bulked guillemot eggs. I single guillemot egg. I herring-gull (Lorux awnfatus) egg. 3 bulked razorbill eggs and 7 bulked kittiwake eggs. PCB was present in all the bulked samples of ail species, it was also present in all the 10 individual sandwich tern eggs and the single guillemot, herring-gull and skua egg. These results show PCB to have been present in all of several species sampled at three widely separated seabird colonies round the British coast. 7lie organo- chlorine residues in four species examined in 1967 on both the east and west coasts were; . Guillemot Razorbill Shat KiHiwaLe Sl AMh Hepd Ifast cm# I ret pp-DDC dieMrto Ippm) ' (ppm) ippm) CKO )t||v Setav 7 etas i 6 3 ) 07 1-0 It 02 01 05 1-0 01 Mutt efCaiioKoy Inc.w C'ti'l) KB pp-DOl dicUrm ippm) (ppm) (ppm) 7 es C|gS 4 eggs bens b 7 5 8 20 M 22 0^7 03 05 1-6 02 KB rtlaltil lo did The terrestrial species examined have been divided into live categoric, on the bam or their main food preferences: bird-feeders: mammal-feeder., carrionbird- and maminal-fcedcrs; and insect-feeders. The amount of PCB present appears 10 vary between the categories. This is Suggested by the ditVerenees in the ranges of residues present (T able 2). the highest levels (70 ppm and SO ppm) being in the bird- and mammal-feeders compared with IS ppm and I ppm in the other inn categories. The time differences arc found in the egg residue figures, residues as high as 10 ppm being found in the 84 eggs of bird-feeding species (merlin 3. long eared owl 4, peregrine 2d, sparrow-hawk 48). up to only S ppm in mammal-feeders (barn owl 4, kestrel 3). and none being recorded in the 23 eggs from species in the carrion-, mammal- and bird-feeding category (kite (Mihvt mttna) 3, golden eagle IS, buuard S). The IS golden eagle eggs that contained no PCB were from inland i I mm (pm f MQNS 086971  tabu: > UV UMOUUOt rnULAKDIUOItl If. II..III.UI Ifl. ltl I, MIIa1n _ fcB I Principal fond (ppll Bints Mwmuh Mammals. Birds Carrion Insects tenge (mie-nux) 0-70 OJO 0-1 No. tpccioiaH 11 7S II Species (with number of each shown m brackets) M*r^* (2) Short-eared Owl (3) Spaow-hawl < |*> Tawny Owl (3) Kestrel <47) Barn Owl (2?) Buzzard (10) Kile (3) Rough-legged Burzard (3) Gofckn Eagle (}) Hubby (|) Swallow (|) LhUc Owl (9) u m!ninjw!'i.... .. HONS 0869 ?<i ' POl.Yt III.ORISAII.I) BIPtll.NYLS IN WILD BIRDS IS Bill I AIN 13 eyries: in contrast an eaglet from North Uist. where the golden eagle's prcv mvludes seabird*. proved to contain 17 ppm PCB. The freshwater ftth-feeding birds examined were mostly obtained from the midlands, east and south-east of England, so in Table 3 the liver residues from these hose been compurcd to those in specimens of resident bird-feeding and mammal-feeding species obtained from the same area. The numbers examined of bird- and mammal-feeders, particularly of the former, are small but the results are of a similar order to those obtained from specimens from these same categories TABLE 3 visia aiwptii or era rilatko to out, is mciMtss prom the midlands, sast and whim or ENGLAND PCB Birth Principal fovd Mammals Fish Range tmm man) xpeeimetu Iftcin (with number of each shown in parentheses) 1-40 0-50 0-m 900 6 18 44 Sparrow-hawk <6> Tawny-o! <3* Barn-owl <l?> Bmem (2) Kioffivhcr (12) Great Crested Grebe <4t Little Grebe (1) Heron tJ?i collected from all parts of Britain (see Table 2). The wide range and very high lev eU found in some of the ftsh-feeders. r.g. ca. 900 ppm in a heron from Hathcrsage in Derbyshire and ca. 300 ppm in a heron from Settle in Yorkshire, indicate thut PCB has apparently been very much more readily obtained and or retained by predatory birds in the freshwater habitats in this area of England, than by terrestrial predatory birds in this or any other part of Britain. 4 between the amounts of PCB ami or^onoch/orme instrllchles A sufficiently large sample of heron eggs and kestrel livers has now been examined to provide an indication of the variation in amounts of the organochlorine com pounds present within one species. Figure 3A shows the residues of dieldrin. PP'-DDE and PCB, the only compounds present in significant amounts, found in 101 heron eggs obtained in 1968 from 6 colonies in east England. Of the three compounds the dieldrin residues are the most limited in range and do not exceed 6*5 ppm. whereas those for both PCB and pp'-DDE extend over a greater range to 80 ppm and 26*9 ppm respectively. There is, however, e larger number of eggs w ith < I ppm PCB (60) than with less than the same amount of dieldrin (42) and pp'-DDE (15k suggesting that of the three PCB is generally present in smaller amounts in (hast herons. X MONS 086973 I 14 UN PRESTT. a I. JEFFERIES. N. W. MOURE A similar comparison of the distribution of residues of the same three compounds in 47 kestrel livers is given in Fig. 3B. Again the dieldrin residues hu\c a more limited range compared with those of PCB and pp'-DDE. particularly as two of the specimens with high dieldrin residues (13 and 18 ppm) probably died from dieldrin poisoning (Prestt ei of. 1968). Unlike the heron eggs, however, there is a larger number of liver specimens with < 2 ppm dieldrin (40) than with less than this amount of PCB (22) or pp'-DDE (23), suggesting that there is a greater ratio of PCB to dieldrin and pp'-DDE in the kestrel livers than in the heron eggs. LARORAIORY EXPERIMENTS ON THE TOXICITY OP A PCS Experimental procrtlure The avian toxicity of one of the polychlorinated biphenyls was tested on the Bengalese finch, a domesticated form of the sharp-tailed finch (Lonchura striata) (EstrMlnae). This bird was chosen as it is easily maintained in the laboratory and V/ W HONS 086979 M l> ntLU IKM \ia* been used at Monks Wood Experimental Station for long-term testing of the MibkthaJ and lethal toxiciiics or pp'-DDT (Jefferies. 1967). ;A proprietary cage-bird rearing food (composed of biscuit meal, dried milk, soya htnn meal and maw seed) vs as used as the vehicle for the PCB. The PCB used was one of the more highly chlorinated typev--Arochlor 1254 (biphenyl with 54** w,w of chlorine). This PCB was chosen as the pattern of peaks produced on a gasliquid chromatogram was similar to that found in wildlife specimens examined at Monks Wood Experimental Station. A 20.000 ppm 'master-mix* of this material in the rearing food was made as follows: 4 g of the PCB was weighed into a foil boat and this was then placed in a beaker containing 70 ml of acetone and the solution stirred with a glass rod. The contents of the beaker were then tipped into a 2 lb Kilner jar containing 196 g of the rearing food ground to a powder (as PCB is a resinous material the difficulty of weighing out a definite amount was overcome by altering the weight of rearing food to the amount of PCB weighed out!. The beaker and glass rod were then washed with two separate 20 ml amounts of acetone which were also added to the contents of the Kilner jar. The acetone was evaporated Prom the rearing food for 10 hours at room temperature, when no smell of acetone could be delected. Finally, the `master-mix' was blended thoroughly by turning the jar on a tumbler mixer for 2 hours. A control diet was treated in the same way. Eight other dietary levels from 10.000 to 75 ppm (see Table 4) were made up in Kilner jars by diluting the master-mix* w ith finely-ground rearing food. Each mix ture was then tumbled for 2 hours A fresh 'master-mix* was made at approximately weekly intervals and all mixtures were stored in the sealed jars at 4'C. A test analysis of 0*9567 g of a S.000 ppm mixture showed a concentration of 5.028 ppm PCB. Thirty-eight adult (>. over 4 months) Bengalese finches, mean weight 14-17 0*1 T |. were placed separately in cages 61 x 41 x 38 cm and maintained at 16 C*. Each cage was provided with cuttlefish, grit water and the standard diet of canary and millet seed. The numbers of birds on each of the 10 dietary levels arc indicated m Table 4. The PCBrearing food mixtures were given to the test birds as only part of their diet. This technique was used to overcome the possible problem of the PCB rendering the food unacceptable to some individuals, which may prefer to starve rather than eat the contaminated food and starvation could obscure any real efleets of the PCB. Thus the standard seed diet was removed from each cage at 14.00 hours and one hour's starvation given before the PCB/rearing food mixture was presented for two hours. A mixture of 15 g of PCB/rearing food with 1*5 g of water was given each time, mixed to a dry paste. At 17.00 hours the standard mcu OKI was replaced. It was found necessary to measure the intake of the test diet, as the consumption varied considerably within each concentration fed. An evaporation control, sei up each day. allowed compensation for this loss and the calculation of the PCB consumption for each bird in ;ig/day (see means in Table 4). With this form of feeding, the actuul parts per million of PCB in the total diet are much lower than that in the PCB/rearing food mixtures given. An earlier series [ MQNS 086975 I li * an rarsTT. n. j. jeFPeaiES, n. w. wnu of text using exactly the same maintenance at various temperatures showed that the relationship between lood consumption ire weight of seed minus husks plus dry weight of rearing foodl. body weight and temperature was given by the equation y J95 50-5'25k (where y - mg consumption of dry food/gram of body weight/ day and a - mean daily temperature in C). Using this equation the concentrations of PCB in the total diet (4 414 g of foodrbird/day) were calculated for purposes of comparison with other work (.ter Table 4). ' TABLE a TUI SANOt or TUT ortrs CONTAINING PCS FID TO 31 lAS0eA10av*MAINTAINID KNOALIU HNCNfS pee 54 DATS as past or runs NOSMAL OtST. thi mkan wiwhcd intabb or rrs ASO rnr CAICUSATSO TOTAL DKTABV UVCL ABI ALSO OlVeN f 11 1 ' to/ dietary /ft*/ (ppmi Control 75 230 300 1.000 1.500 2.000 3.000 10.000 20.000 Sombre nf bird* turd $i _6 . 221 21 41 421 21 4I 4- Mean PCB canumptba 0 26 73 125 217 367 502 1.011 1,3)1 1.040 Calculated fatal dietary level (ppm) 0 A 17 2* 49 S3 114 229 302 440 The 3S birds were maintained on the above test diets for 8 weeks. Those still $ aiisc at the end of this period were killed with ether except for 5 test birds (73-2.000 ppm diets) and 3 controls which were starved for 21 hours prior to being killed. &. This was done in order to evaluate the amount of PCB released from the depot fat into more sensitive orggns when a bird it under stress, at in bed weather or after long flights or illness. All birds were examined and the brain, liver, kidneys R and heart weighed. The livers of all test birds and the brains of most were analysed a** for PCB concentration. ' Hr Avian loxiciiy of PCB Seven birds died during the 36 days on the test diets of PCB. Deaths occurred at 17. 18,21. 26. 32,40 end 42 days with no apparent correlation of time with dose rate. No mortality occurred in the control. One of the earlier symptoms of PCB poisoning was an apparent lack of balance at perching. Later three birds which died showed a form of paralysis in the legs. with one leg carrying all the weight whilst the other was splayed out to one side with foot closed. Before death some birds showed body trembling with shaking wings. At the consumption of PCB varied considerably at each dietary level, the birds were regrouped before calculation of the LD30 according to their individuel " . `' \; 1. [ ; -I ;u pm ` MOMS 086976 POLYCHI.ORJNAUI) HIPIIINYLt IN Wil l) HIKIlS IN HNIIAIN IT consumption in mg PCB kg body weight day. Five dosed group' were used besides the control. These were 0-12. 12-JA. 3A 72. 72 144 and > 144 mg kg day, The percentage survival in each group, together with meun dove rate und number m the group are shown in Fig. 4. None of these dose rates produced 100 .. mortality. Tk* hi|kni mortality, one out of two birds V.U. shown at the highest dose rate O: 6 b: 0 C: 0 10 6 52 9 38 2 24 55 101 171 mg/kg/doy 107 161 218 283 ppm r if . Fig. 4. The prromaae suni'al Mhn each of ihc five kne rate grout* and control <al number* in gaeb group; (b) maan deae rate in each group and the calculated mortality curve. The calculated liver concemratiom in ppm wet weight are alio shown (cl. with a mean of 170-7 mg. kg day The factors limiting the production of higher mortality figures were the low toxicity of the material and the apparent unaccept ability of very high concentrations in the food. There was a significant correlation between the concentration of PCB in the food and the amount eaten <y - 0-5327 IMH75JU where x log ppm PCB in test diet and y w grams dry rearing food MONS 086977 IX IAN PHCSTT. D. t. JCFFSKIEA, N. W. MOOAI ! > eaten per day; r -0-6581; DF 30; P < 01%). Thus as the concentration in the test diet increased, the daily consumption decreased. Increasing the top dietary j- level from 20,000 to 40.000 ppm would not have greatly increased the daily intake , of PC B. The mortality curve and regression equation were .alculated by probit analysis ' tire Fig. 4). The equation was found to be y 2-8826 + 0-8807* (where x - log I dose rate in mg/kg/day and y - percentage mortality m prohits) and the estimated U>50 56 days was 2-4042 0-3765 log units (* 255-6 mg/kg day or u total of 14.200 mg/kg for the 36 days). Estimated dose rates lor 75% and V9% mortuhty ; using this equation are 1.462 and 112.100 mg/kg/day respective!). These quantities would he difficult to achieve using the present feeding technique [ Of the five starved PCI* birds, one died'afler 20 hours and a second after 2! \f hours starvation. At this time the remaining three were on their cage floors showing . signs of stress and two were trembling; all starved birds were then killed with ether. , The control birds were perching and one was flying normally after the same period of starvation. ` : There is very lutle published work on the avian toxicity of the PCBs with which r to compare these findings. Flick et ai. (1965) fed Arochlor 1242 at levels of 200 and - 400 ppm in the diet to one-day-old white leghorn cockerels in order to compare the ; response with the svmptoms of chicks suffering from a new pathological disorder - known as 'Chick Edema Disease' (Schmittle el at.. 1958). Unfortunately the ; Arochlor was fed for three weeks only, by which time three out of 24 birds had : died on the 400 ppm diet. This result is somewhat similar to that of the present experiment, where three out of the seven birds which died on diets up to 440 ppm had done so by three weeks. Koeman et ai. (1969) fed a diet containing 2000 ppm of Phenothlor DP6 to 20 Japanese quail (Cotumix coturnix). all of which died in 55 J days. This total dietary level was not achieved in the present experiment but. using the data in Table 4. an approximation would suggest that this might have resulted in an intake of 10.000 /<g'day. Figure 4 shows that this would have produced a similar, almost complete mortality in a similar number of Bengalese finches. * , 1 Boitv it eights. organ weights ami post-mortem findings During the 56-day period of the experiment the controls increased in body weight by mean of 3%. The 20 dosed birds killed at the end of the experiment also had a net mean gain (6 %). Birds dying of PCB poisoning had a net mean loss of 13 which was similar to the mean loss of 14 % shown by the birds which were starved for 21 hours. The weights of the fiver, heart and kidneys of each bird were related to its brain weight (mean 0*453 0-005 g; this forms a better indicator of body size than the more variable body weight) by calculating the percentages organ weight/ brain weight .x 100/1. The liver percentage wu similar in controls (139-5%). PCB birds killed (136-6%) and birds dying of PCB poisoning (143-0%). However. ' \ l I MONS 086978 V n&>C'm.Oft)NATKD MIFIM.NYLS is WILU MMIft IN IlMII MS } )V n PCB binlv tlurvcU for 21 hour*, the liver percentage was considerably decreased (i presumably due to the loss of organ fat. The heart percentage was lightly increased from in the controls to 40-9*,, m the PCD birds hilled and o 4441** In those dying of PCII poisoning. Two of the males which died hud <rt percentages ol 51 and 52",, which are outside the range of normal heart terceniage* for males of this species (Jefferies. 1969). The kidney percentage was imihir in the controls (52-4*,,) and the PCD birds killed (JO-#",,) hut was very much irjftr nt the birds which diedf.VM"*). There was probably a slight reduction in the ulncy weight of the starved birds (28 The most obvious gross pathological change seen on post-mortem examination 4 all the birds dying from PCB poisoning was the enlargement of the kidness. slbO. two of these birds showed considerable amounts of fluid in the pericardial ac. Hydropericard* were mentioned by both Flick ft al. (1965) and Kocman t ct. (1969) in chickens and quail poisoned with PCBs. Flick ft al. also found onsiderable kidney enlargement. Very little depot fat was found in the birds tying from PCB poisoning and none in those starved for 21 hours. CamctMraiion of PCB in the organs of living and dead birds The brain and liver of one of the control birds were analysed at the end of the experiment and no PCB was detected. * The concentration* of PCB in the livers of ihe 20 birds killed immediately alter :he end of Ihe experiment ranged from 3 to 6.14 ppm. that in the seven birds dying men PCB poisoning from 70 to (97 ppm. These concentrations are shown plotted gainst dose rate in Fig. 5. The eoncentrations found in birds killed und birds lying overlap considerably. There was a significant correlation between the dose Mt and the concentration of PCB in the liver (y - I 336 + O-JOOx. where log PPm PCB in liver and x -- log dose rate in mg 'kg day: r * 0-5414: >F 23: P < I *,,). Using this equation the liver concentrations for each m the low rate groups were calculated and are shown in Fig. 4. The variation in the ir PCB concentration at any one d>c rate is extremely large (the 95 confidence anils art 0-9490 unils of y above ami below the regression line--e-.e- for the U730 4255-6 mgkg'day the liver PCB concentration would be 24? ppm with limits from 9 to 3.070 ppm). This may be due partly to variation in fat content since Ihe liver ooctotration in Ihe starved birds, as well as being much higher (69 to 1.214 ppm) .as not so variable (are top of Fig. 5). Howeser. the variation is too great for this 4 be the sole cause. The relationship of the concentration of PCB in the brain to that in the liver at taimined for each group of experimental birds by calculating the percentage, min ppm'tiver ppm x 100/1. In the PCB birds killed at the end of the experiment : was low. a mean of 26-4 6-1 whereas in those dying from PCB poisoning : was over three times higher. *3-7 7-4% In the starved PCB birds the percentage at between the two at 47-6 7-3%. I 1 V m TO WWW'WPTfi M0NS 086979 m m n yu.ni 20 IAN NUMT. I). J. JEFFraiCII, N. W. MOOnr One bird (15 04 g), fed 1.500 ppm PCB and killed at the end of the experiment, was submitted to total body analysis to find the percentage storage of the PCB committed. The brain (0:42 g), liver (043 g) and depot fat (0-23 g) were analysed separately. The rest of the body was weighed (12-20 g) and analysed together. The K* total body weight was made up by gut contents (0-84 g) and feathers (0-72 g). ft PCB content of the brain (tpg: 14 ppm), liver (16 pg: 25 ppm), fat (132 pg; 574 ft MOhlS 086960 jpw w i i g ifm p p B jiip g w p tLii w n i, j i t >"* KM.V(*MU)IUVAltl> HIVIIINYI.S IN WII.IS Him*, IN HHIfAIN 21 ppmI and rest of body (750 //g: 01 ppm) wax 904 /if. This bird consumed 10.24k .if of PCS and so uored 9", of the PCS it hnd eaten. It it stored in the depot and orfan fat of the body. Cotn/turiMnt nf the lo.xicit\ u) l`CB with that of arganm Marine linn ucitk'y Lonf-term experiments on the sublethal and lethal toxicity of DOT and us netaholitet hate been carried out using this species with the iameesperinicni.il chnique. Although nnt cnmpletcd. (he LR 50. 56 days of pp'-DDT has been fouiul to he approximately 1100 mg kf (19 mg kg,day). Thus the PCS Arochlor 1254 has .mis I 'll of the toxicity of DDT (DDT in turn has only I 6 to !/M of the toxicity of dieldrin. De Witt rr at. I960). However. whereas DDT has a steep mortality curse il.e. no deaths at approx. 1000 mg kg. oil dead at 1200 mg kg using 122 birds: 11 and 21 mg/kg day respectively >. this curse is much more gradual is ills PCB. This means that at low dote rates, below Ik mg kg day. PCS mas base a .omparatisely greater toxicity than similar amounts of DDT (i.r. to the 2-10"., fihe population which is sensitise to these low amounts). Dtlrrlbg mortality from PCB poisonirtff On intake of PCB this material accumulates in the organs of the hods. particularly Ihe depot let. in concentrations commensurate with the dose rate. It would appeur that death follows enlargement of the kidneys and hydropericardium. The liter itmains normal in tiae at death as the turd dies before the organ and depot Tat have heen completely used. Some body weight lost occurs, possibly due to lack of feeding prior to death. The PCB from the depot fat which is used at this time appears to go to the brain and increases the quantity there considerably. When a bird containing PCB starves (as would occur in the held during bad weather or illness) the same thing happens and the brain concentration increases. However, the concentration in the liver alio rises due to the reduction in liver weight with Ion of organ fat ithc highest content of PCB in the liter of a bird which died was 740 /ig. 697 ppm the highest content in a starved bird was similar at 670 /ig although the concentration had risen to 1214 ppm). - With materials such as DDT where the correlation between dose rate and liter content is very good (Jefferies It Walker, 1966) and the mortality curve has a steep slope, an indication of whether a bird has died after poisoning from that compound may be gauged from the concentration in the liver. With PCB. however, the opposite is the case. There is a hundred-fold variation in the liver concentration at each dose rate and the mortality curve has a gradual.slope so that over a considerable range of dote rales the lethal range of liver concsggrations is paralleled by a similar range of non-lethal concentrations. For example, the highest lethal concentration in the present experiment was 697 ppm hut one bird on a similar dose rate lived with 6.t4 ppm in the liver. Thus there it no certainty, given the liver residue alone, whether a W l V pi rm MONS 086981 t t 22 ian mfstt. i). j. Jtmsrfcs. m. w. worse particularly large concentration has caused death or not. If of a similar sensitivity to striata, one in thirty birds would die with liver residues from 5 to 360 ppm (mean 40 ppm), one in three with 22 to 1780 ppm (mean 200 ppm) and one in two with 39 to 3070 ppm (mean 343 ppm). It is unlikely that death would occur at amounts much lower than S ppm. A more certain'conclusion about the probability of death from PCB poisoning may he reached ifthe brain is analysed as well as the liver (to obtain the relationship or the liver/brain residues) and the body it given an autopsy paying particular attention to enlargement ot the hydropericardium and kidneys. CONCLUSIONS AND DISCUSSION The majority of our samples tested Tor PCB have come from predatory birds. Studies of organochlorine pesticides have shown that larger residues of these compounds are present in predatory species than in omnivorous or herbivorous birdsl Moore A Walker. 1964: Prestt, 1967). The organochlorine pesticides and the polychlorinated biphenyls arc similar in several ways. The presence of PCB in such a large number of specimens from diverse habitats throughout the country, in relatively large amounts, suggests that they, like the organochlorines. are persistent. Also, both accumulate in the body fat of animals. Thus one might eapect that predatory animals at the lop of food chains would also be the best group to indicate the presence and the geographical distribution of the PCBs and to suggest whether or not they represent a harard to British wild birds. Other studies (Ritebrough tt a!.. 1968; Koeman rr at, 1969; Jensen et a/.. 1969) have suggested that residues of PCBs. like residues of certain organochlorine pesticides, may now be widespread. Our study confirms that they are widely dietributed in Britain. In the terrestrial environment they were present in birds and their eggs from all parts of the country. In seabirds they were found in all species sampled al three coastal colonies, one ofT the Irish sea and two o(T the North sea. The largest amounts discovered were in lake and river birds from the midlands. east and south of England. The highest of these, ca. 340 and ca. 900 ppm, are the largest liver residues of PCB yet reported in wild animals from any part of the world. Like pp'-DDE and dieldrin, PCB it present in larger amounts in the birdfeeding and fish-feeding species. The highest levels found in Swedish waters by Jensen rr al. were In the fish-feeding white-tailed eagle and heron. The level of PCB found was also similar to that of pp'-DDE. The average residues of dieldrin, pp'-DDE and PCB for four predatory species obtained in Britain from 19(3 to 19(8 are given below and show this similarity (data for pp'-DDE and dieldrin, from Prestt in press and for PCB from Table I of present paper). 5 ? 'r i' f ' t l> J f ' ; > j ' j k j t ! : . F POt.VCHlOIMNATt.ll ail>lll:NYU IN WILD MAIMi IN HNIIAIV 33 KCttfCl HftTfW*! |iir N. v 42 57 13v 102 pp'-DbC (ftpm) 30 II 14 5 so 42 titrMri* ippm\ 933 44 VI . 24 included in each |nup n r Table* 2 and f rc%hair iKh^ccden* RiriMcctfcrs* ' MammaUtedcft" PCM tppmt 40 10 4 In view of Ihc Similarities or the distribution and levels of PCB to those of pp >DDE and dicldrin. the immediate question is whether PCB could have caused Ihc effects on predatory birds in Britain previously attributed to the organochtnrine pesticides. As far as lethal poisoning is concerned, there are several reasons fnr considering this unlikely. First, our toxicological studies show one PCB land comparisons with the other published toxicological studies suggest that Ihc tntKitiet of the different PCB compounds may not vary very greatly i to have only about I I) the toxicity of pp'-DOT. DOT hav only I A to I 14 the lethal chronic toxity of dicldrin (De Will rr a/.. I9A0| and our studies have shown that pp'-DDE has an even lower lethal toxicity than DOT to the Bengalese finch. Very frw of the specimens examined are likely, therefore, to have died from either PCB (or DDE) poisoning. As there is no reason to suppose a greater use of PCB in Britain prior to the period during which the present samples were obtained, it i< unlikely it has ever had any widespread acute elFects in Britain. Secondly, w hilc the population crashes of the peregrine and sparrow-hawk were coincident in space and time with the introduction of aldrin. dicldrin and hcptachlor. i.e. in the mid IMOs (Ratdilfe. I96J: Prestt. 1905). the introduction and use of PCB is not. PCB was first used in 1930 and, while it probably became more available to wildlife xfttr the I9J9-45 war, there it no reason to suppose it might suddenly have produced in exceptional effect in the late 1950s. In the same way. while the recovery of the yarrow-hawk and peregrine started in the mid I9t0s following certain bans on aldrin. dicldrin and heptachlor. there is no reason to suppose there may have been a reduction in the amount bf PCB available at this time (it should be remembered dial the dicldrin figures shown above were obtained after the bans). The geographi.91 distribution of PCB appears to be general in the terrestrial environment in Britain, to in this way also there is no connection with the population declines w hich were more severe in the south and east of Britain. Nothing in the ev idence available shout PCB. therefore, contradicts the previously advanced hypothesis that lethal soitoning by aldrin, dicldrin and heptachlor is the most likely explanation of the udden declines in certain birds of prey in Britain in the late 1930s (Jefferies A retit. 190b). As regards tha sublathil effects on bird populations the situation may be different, "here has been a widespread breeding failure among predatory birds in Britain i I ......nmriii. HONS 086983 .'4 UN .'HIM I. It. I. II I linns. N, W. VM*.,,, ,mhI the ks\ in pox w.u years t KaicMlc. iW.t, 1`JMa: l.uckic & Kunlislc. WM. IJukey. \Um mans speues (again particularly ihc predatory buds! have been shown n Ik InNing iluitshelled eggs miicc appnMinatcly f>47 (Kaicmlc. |Vf"h. Make* rt Anderson, laboratory cvjvrmicnls hero show (hat ino rcpiodiivtivc \stcm '! hiuls can lx* adversely aliened by orgunochlormes such as |>|>I iJoll'oMfs, l%7. i'tb'i. I\ ^m, K5M and the same compound has been slum; ccpernuem.dU ( cause lho production Ol` thllt'Shcilcd Cggc ill the American kostiol fufttt ywefMll'itfier K Wicmcycr. K6V). Kcccnt work by Pcakall (iw'i has shot*n that one of ihv mevhumxm by which this mu> he brought about is the induction >!' steroid by dross bung cn/y mes in ihc liver (see discussion in Anderson ft ,tl, As K B is lifts known to produce a similar reaction(Kischrough i7 at., lOhKui H'oms possible that P( II may he one of the component causes of the present hiceiling failure. This breeding failure is probably an important factor advciscly aliening ihe recovery of the bird of prey populations. Andersttn vi a! in their study on the circus of orgunovhlonne compounds on breeding pelicans I/VA-mv/m* rrrthrurhsnttts) and cormorants (fhalutrtHoru.x attrnus), reached a cmiuI.m conclusion that there was no evidence of acme loxiutv from K.'B 01 pp |>|)| . the two mam t impound* present. bin both correlated with change in eggshell thickness H ilt are produced and marketed under a number of commercial trade names re \rothlor. Clnphvn. .md arc available a> liquids, resms or solids 'they .oc insoluble m water, but have a low timic vapour pressure 1 hey already have many mwn: protcilivc coaling", plasticisers, sealers m waterproofing, printing inks, synthetic adhesives. In liquid form they are used as hydraulic fluids. m thermostat*, culling oils: giHiding thuds and they are incorporated into electrical apparatus there are thus many ways m winch they can become widely dispersed in ihc environment into the atmosphere with indiistrud smoke, or in exhaust from air* waft engines amt into rivers an factory effluent. It has even been suggested they may have been incorporated in pesticide formulations to increase (he persistence of insecticides (Reynold*. I*>W). In view of the large number of dilfcrent uses of KH ami its apparent persistence, it i* not surprising that it is now present in a large number of wild birds. Moreover, as it is an industrial pollutant, its control will present an entirely ddlVreni problem to that of the control of the organochlorine pe*<wide*. I i- r i I ACKNOWLEDGEMENTS We are particularly gruteful to I). C. Holmes and the staff of the Laboratory ot' the (iovernment Chemist in London, and M. C. French and P. Freestone of Monks Wood I sperimental Stution. for currying out the chemical analyses. The specimens were kindly sent by interested members ofthe public, many of whom are members of T*- *ott$ 08698, t' <>k* Royal Society lor the Protection of Birds and the British Trust for Ornithology. V A. Hall. A. Jackson and T. Moss provided assistance in the laboratory and D. A. itatcliffe and A. S. Conke kindly commented on the manuscript. ttittaiM is ^Vt'Kv.v |> >k.. IIWAIS. J. J,, KlMHNOl <!. K. W,, tin.HIS. I). f\ A OlNIMiNMN. K. I . 'S>|ii>ilk'HiKc oi chirr tn.ia-tl n)drrcirt>ofl rci^Uluc^ to breeding PclN'tn* ,mtl l .wmt*uniik.' I'm. IM. Air/.. M3. VI 112 i i mii Mai \mMs M, j,, I.i.as. II., (ioniv, li. W.. Hammond, L. W., Komi an, J. A |hmsos, j. H9M). `Clean up of annual Mis and dairy produce for the analysis ofchlorinated lA/Miffd* residues,' Lnnd.. If, lhH-74. It Mot. 3 R.. Mn/h. C. M.. Aiwimii*. V. A. A Rucmm. W. L. (19*0). 'Pcsticidal residue* in animal hsmiCv' Trmt. A. .4*i. Il Ml. ( //. 25ih, 277 V). h Win. 3. II M953). bffevts ofchlonniied hydrocarbon insecticides upon Quail and Pheasants.* J wffW hi Ottm.. . *72 ft. ima.17 I O'Oiii, R (i. A Ciiiiih. V. A. il9Mi. Studiesof the chick edema disease. J.Jmwi* Mint. iM stmptttms produced by feeding chlorinated biphenyl.* PiMt. 5c/.. 44, In mis. J. J A ANP*aw>v O. W. iIVAN). 'Chlorinated hydrsvarhont and eggshell changes in ..ipMrwl and ftsh-eaung Nrdi,' Seiemt. A 1*2, 271 - J. it>Mi>, J I ltd 1 (19*9) rrrfgrim' fr/ttmi P>'ptthifon\. Wisconsin; Unis, of Wucontin Preu. lot mi a, r> <*, Sisimhn*. J. 31. A Tstiov J. OO. 11967). 'Chlorinated hydrocarbon* in British wildhfe ' Wmcc. Cm!.. 216, 227. Minna. D J. A farm, I. H966). Pothmoncm* of Peregrine* and Lanncrs with particular whwiM to tsrumschWtnc residues.' Br. Birth. 99.49 M. MfMttfK, K>. J. k WMMa. C. M. (1966). 'Uptake of pp .POT and its poM>mi>rtem breakdown m the as tan User.' Xmm\ Lmd.. 212.332--4. Minna, f). i. H96t). 'The delay in ovulation produced by pp'*OOT and ns possible significance ni the Meld.' M/i. lit. 266-72. D. 3.11969). induction of apparent hyperthyroidism in birds fed DDT.' Smurf. L.*hI,, Itt. 579.9. %M\, S. (I966i. 'Report of a new chemical hazard.' AVh Seifhi., 32.612. rSMH. . Jummia. A. O.. Otnov M. k Oturmno. C. (1969). 'DOT and PCB in marine annuals from Swedish water*.' Ve/nrc, Lomf.. 224. 247-50. MIMAS, J. H.. Tin Mmsti DC Brawsa, M.C.ket Vok, R. M. (1969). 'OtlorinaicJ Biphenyl* In mb. mussels and bird* Rom the Riser Rhine and the Motherland* coastal area.' \atuu . bmt.. Ill, | I26.g, trait, I. D. A RAtct-tm. D. A. < 1964). `Inicetieide and Scottish GoiMn Eaglet' Br. Bints. 57. blunt. S. W. A Waikir, C. H. (1964). `Organic chlorine insecticide rctidue* in wild bird*/ 1 pmit. 1072-3. ****. V W. A T*nrs, J. O'O. (1965). `Organochlorine miccticKk residues in ihe egg* of sea bud*.' VoTart*. Umt., 207,42-3. Mirn. R. D. A Wiinivui. S. N. (1969). 'DicMrin and DDT: effect* on Sparrow.hawk eggshells and igprodwcthtn.' Srimri*. \. Y., 165.199-200. *mi. I. (1962k 'An enquiry into the recent breeding stntu* of tome of the tmaBer bird* of prey and crow* In Brttein.' Hut StuBy. 12. 196-221. teu. 1.0966). `Studies of recent change* in the Matos of some bird* of prey and fob*feeding bird* m Britain.' J. appI. rf., 3 (Suppi-k 107-11 uait. i. H967), 'InveMigaiiont into poMibie effect* of organochlorine inaectieidM on wiki predator) bird*.' Per. Br. Imrefif. Fm*k. Omf.* I, 26-357 rr. |,, JrtitRHt, D. i. A MAroouAt.o, J. w. (1961). 'PoM-moriem eaamination* of fswir Mownh lmged Buuard*.' ffr. fftoA. 61.437-65. Mar*, t. tin pm). `Organochlorine pollution of river* and the Heron (Arrfra eAtmti L.)' Tech. Mm. im. Un. Conaerv. Not. nai. Retnur.. li. New Delhi. ATCurrr. D. A. (1962). The Matu* the Peregrine in Creat Britain.' Bint Stmly. 10, 56-90. ttttim. D. A. (|967). 'The Peregrine ahueiion in Greet BrMain 1965-66.' Bint Study, 14, 26 ian mtsTT, o. i. jerraiuEs, n. w. moors lUrcum, D, A. (19*79). `Decree** m eggahell weight in certain birda of prey/ Mature. UmH.. mjoi-to ** * JUrNm.oe, L. M. (196V). 'Poiychlorobiphenyk (PCB) and the tmerferenee with pMtietde residue anotyal.' Bull. n>. oenMm. A Toxkitl. (C/5X 4,I2M3. . RlMMOUOM. R. w., RorHl, P., PtAKALL. D. B.. Human, S. G., * KiavtN, M. N. 11961). `PotyeMorinataid biphenyls in the global acoaystam. Nature, Lorn!., 229, 1091--102. Rm-iN. J, 11965). *A simple concentration cell technique for determining small amount! of beild* ions and Ms use in the datarmination of raiidun of organochlorine pesticide*.' . itmfyst, ICNMrrvu, t. C,, EowAaoa, H. M. A Moans, O. (I9SS). `A diaordar of ahiakana probably dua id a low fcid--preliminary report.' J, Am, mi, mti. Au* US, 216-9. B I MQNS 086986