Document qd6d1wKa0EEnz7XGkkmoB0z9x

, . - Paf+ 'V* / Polychlorinated Biphenyls Another Long-Life Widespread Chemical in the Environment David B. Peakall and Jeffrey L. Lincer The recent finding that pelagic birds dying on the coasts of Great Britian had polychlorinated biphenyls (PCBs) in their livers in concentrations of several hun dred parts per million (Bourne and Mead, 1969) shows that these compounds are present in the ecosystem in large amounts. Thus, it seems worthwhile to summarize and evaluate our current knowledge of these compounds. Structural and Physical Properties The picture is complicated by the fact that we are not dealing with a single com pound. The basic structure of PCBs is shown in Figure 1. Any of the positions marked with an x can be substituted by chlorine. Widmark (1968) has calculated that of the 210 possible combinations 102 are probable. His criteria for these lim itations are those compounds containing five to eight chlorine atoms per molecule and the number of chlorine atoms per ring differing by not more than one. The commercially available Aroclors (Monsanto Company Trademark) are des ignated by numbers (Monsanto Technical Bulletins). The first two digits represent' the molecular type: 12 - chlorinated bi phenyls; 25 and 44 - blends of chlorinated biphenyls and chlorinated terphenyls (75% biphenyl and 60% biphenyl, re spectively); 54 - chlorinated terphenyls. The last two digits give the weight per cent of chlorine. Thus, Aroclor 1242 is a chlorinated biphenyl containing 42% chlo rine. The biphenyls commercially avail able from Monsanto range from 21% to 68% chlorine. Mass spectrographic studies of Aroclor 1260 (Koeman et al., 1969a) show the presence of 11 isomers: five con taining six chlorine atoms, five containing seven chlorine atoms, and one containing eight. Bagley et al. (1970), studying Aro clor 1254, found 18 compounds: one con taining three chlorine atoms, four contain ing four chlorines, four containing five / chlorines, five containing six chlorines, and four containing seven chlorines. Thus, Ttic auihort are currently at the Langmuir Laboratory, Cornell Uiuvenity, Ithaca, N.Y. the number of compounds present is, for tunately, much smaller than is theoretical ly possible. PCBs are chemically inert, are not hy drolyzed by water, and resist alkalies, acids, and corrosive chemicals. They have low volatility, their boiling points ranging from 278 C for Aroclor 1221 to'415 C for Aroclor 1268 (Penning, 1930). All are stable to prolonged heating at 150 C, and the lower Aroclors can be distilled at atmospheric pressure without appreciable decomposition. PCBs are described as in soluble in water and very soluble in hydro carbon solvents, although no exact figures appear to be available. Nothing is known about the biological decomposition of PCBs, but it is likely that they are more stable than DDT and its metabolites since they lack the ethane component between the aromatic rings, which is the site of action of most of the transformations of DDT. Thus, PCBs have'the necessaryphysical and chemical characteristics for persistence and accumulation up the food chain. Use of PCBs Polychlorinated biphenyls were first de scribed in the literature in 1881 (Schmidt and Shultz, 1881); and the successful com mercial production was achieved by the Swann Company in 1930. In that year the physical characteristics and commerical possibilities of these materials was de scribed by Penning (1930). PCBs are now manufactured by Monsanto in the United States (Trade name Aroclor), Prodelee in France (Phenochlor), and Bayer in Ger many (Colphen). Other manufacturers are located in Japan and the Soviet Union. No figures of the amount of these mate rials produced annually are available, but to judge from the uses listed in Table 1 they may be very large. The uses listed in the table are only suggested applications taken from Technical Bulletin 306 of the Monsanto Chemical Company, and again ' there is no information on the extent to which these recommendations are acted upon by the manufacturers of plastics and resins. The use of highly chlorinated Aroclors for extending the kill-life of formulations containing chlordane,,aIdrin, and dieldrin are mentioned under miscellaneous appli cations of Aroclors. This possibility has been considered in a number of papers. Sullivan and Hornstein (1953), Hornstein and Sullivan (1953), and Tsao et al. (1953) found that the chlorinated terphenyl Aro clor 5460 increased the residual persis tence of lindane. Tsao et al. (1953) consid ered that there was some indication that chlorinated polyphenyl may have a syner gistic effect with lindane. Duda (1957) PCS xx % x> x X XX X X POSSBLE POINTS FOR substitution of chlorine (A) oio Cl-C-CI aI DOT DKHLOROOtPMENfl. Fig. 1. Basic structure of PCBs (Al and possible reactions of PCBs (Bl. DSW 552533 STLCOPCB4090950 sprayed elm saplings with a lindane Aroclor 5460 mixture (1:4) and then tested the residual effect on the leaves to infes tation with elm leaf beetle (Galerucella xanthomelaena). He found that leaves treated with lindane plus Aroclor were protected longer and that the insecticidal effect was more rapid than with lindane alone. Peaks corresponding to Aroclor 5460 have not been detected in the envi ronment. However, under conditions of temperature and flow-rate commonly used for gas chromatographic analysis these compounds would not be detected (Reyn olds, 1970). Nearly 25 years ago it was noted that Aroclor 1242 was one of 175 compounds out of 6000 tested that was effective against mosquito (Aegyps) larvae (Deonier et al., 1946). Lichtenstein et al. (1969) tested the effect of adding a variety of biphenyls and terphenyls to DDT and dieldrin with respect to their toxicity to house and fruit flies. It was found that PCBs had very low toxicity to house flies when given alone but PCBs increased the toxicity of dieldrin and DDT, especially the latter. The effectiveness of these com pounds decreased as the chlorinated level increased. For example, Aroclor 1224- in creased the mortality of fruit flies from 59% to 93%, whereas Aroclor 1268 in creased it to only 77%. Analytical Methods 1) Identification. Since various com pounds with electron-capturing properties have been tentatively identified in atmo spheric samples in the past (Abbott et al., 1966), and some pesticides are capable of hybridizing in the soil to form a new com pound (Bartha, 1969), it is important that the presence of PCBs in field samples be proven beyond doubt. Identification by means of a combination of high resolution gas chromatography and mass spectrom etry has been carried out by three inde pendent laboratories in Sweden (Widmark, 1967), Holland (Koeman et al., 1969a), and the United States (Bagley et al., 1970). Widmark's report states that all peaks were identified by mass spectrom etry (although no data were given), but Koeman and co-workers gave full experi mental details, and Bagley et al. (1970) demonstrated that most chemicals in the eagle samples examined were components of Aroclor 1254. Thus, despite a recent statement by the Monsanto Chemical Company (in Risebrough, 1970) that the case for labeling the peaks in question as PCBs was not proven, there is enough evidence to convince an unbaised scienti fic jury. September. I, 1970 2) Separation. The chemical techniques preliminary to quantitation of residues in samples containing both PCBs and chlo rinated hydrocarbon pesticides fall into two groups--those necessitating the de struction or alteration of one or more of the compounds, and those which do not. Included in the first group is nitration. Treatment with a 1:1 mixture of sulfuric acid nitric acid at 0 C for 5 min destroys or alters aldrin, p,p'-DDE, p,p'-DDD (TDE), p,p'-DDT, and dieldrin such that they can no longer be detected at the original position on the chromatogram. This procedure leaves unaffected PCB, lindane, and BHC (Jensen and Widmark, 1967). A more rigorous nitration with a 1:1 mixture of sulfuric acid-fuming nitric acid for 15 min at room temperature re moves, in addition to DDT and its related products, aldrin, heptachlor, Kelthane, Perthane, Tedion, Telodrin, and Trithion while lindane, heptachlor epoxide, toxaphene, and Strobane are not removed (Erro et al., 1967). Risebrough et al. (1969) reported that this nitration also removed the chromatographic peaks of PCBs. Rey nolds (1969) reported that his attempts to nitrate samples- were not fully successful in that there appeared to be loss of some of the more volatile PCBs while peaks with longer retention times appeared. Armour and Burke (1969) reported that complex chromatograms resulted after nitration which could not be related to the unreacted DDT-PCB mixture, and nitra tion was not pursued as a practical means of separating DDT and PCB for further' tests: Saponification with alcoholic NaOH or KOH will dehyrodrochlorinate Perthane, Toxaphene, DDD, and DDT to their re spective olefins (Archer and Crosby, 1966; Klein and Watts, 1964). Risebrough et al. (1969) reported that PCB peaks are not removed or displaced but gave no data. The second, and in some instances more desirable, group of analytical tech niques allows the special separation of many chlorinated hydrocarbon pesticides from PCBs. Reynolds (1969, 1970) re ported on an activated Florisil column technique which separated heptachlor, aldrin, DDE, and PCB with the first elu tion (60 ml n-hexane) from lindane; hep tachlor epoxide, DDD, and DDT with the second elution (40 ml 50% ethyl ether in hexane). Armour and Burke (1970) devel oped a method utilizing a silicic acidCelite column eluting aldrin and PCB with the first fraction (250 ml petroleum ether), and lindane, heptachlor, hepta chlor epoxide, dieldrin, endrin, p.p'DDE, o,p'-DDT, p,p'-DDT, and p,p'DDD with the second fraction (200 ml acetonitrile hexane methylene chloride-- 1:19:80). Koeman et al. (1969a), using an activated Florisil column, eluted the apolar compounds including DDE and PCB with hexane, and then dieldrin and endrin with 10% diethyl ether in hexane. Mulhern (1968) reported on a method which utilized silica gel-coated thin-layer plates Tabu >. Im m d pafychlariMta4 bipkearfi Hit* ha* Monuata Tachnicd Bvflaba 0/U-ttl lAatarid with which Atachr it aibiMi Aradar atatf . srt* at % Cm Nytiarf ctferdi iiinAiiit Wtytn antat* fthihai iayt ' KIWI i*Mt * N|Ula taaaa Arodar 1241, 1254 1 12U (l-*M Mm 122 t7%) Atadar 1221, 1222. 1242 (11%l . Aradar 1254 (41%) Aradas 1221 & 1240 0*%) Arad* 12M (10-15%) Aradar 12H (10-20%) ItMdatt fUttidnt ta nprava Rasa rattrtaata mi cheated raatuacc. . Cfrptudcuai ti twhsac* rsd-rtf. iaprtv* VMfc-tck md . fhar-tear papsitits Praaura-itftativt adtaara. iacrusa Chawed id - . adatisa ratiostea aari ' rfwiva pattat. ' Pftctrrc md atatawed Art rturfeac. hciaasts raanyth af har-^an raiofacad rasa. OBariaataA rabhar On if hitadiaa aapafrf Crf IWUar nht ,, '' Atadai 1221 (2%) *fadw 1254 (5-11%) Mm 1254 (tig Mm 12U (40%) Aradar I2U (1.5%) Aradar 1212 (S-$0%) Mm 12U (25% af MO Mm 1242 11%) ftttbdtsr. (diwii xstiim. Rasa . ratsAaaca. md iaprrrta daittird ifUtil gap artist. dpam chawed rasbunca. .. fir* rturlaat, ItjacBaa sddn|. flatecuar a pant - fpaatiant. Sapravci mi rttittaaa Msitai* ad Raa rasitiag , DSW 552534 959 STLCOPCB4090951 and a hexane/ethyl ether <98.2) solvent fact many of the original samples con system. The plates were developed, tained little or no p.p'DDT, it was pos sprayed with a silver nitrate solution, and sible to estimate relative PCB values. exposed to UV light. The plates were then Anderson and his co-workers also saponi divided into five horizontal sections. Di- ' fied samples to remove interfering p.p' eldrin, endrin, y-BHC, heptachlor DDT and p,p'-TDE and then quantitated epoxide, p,p'-DDD, p,p'-DDT, o,p'- PCBs as Aroclor 1254 by relating sample DDT, and p,p'-DDE (in that order) were peaks 9 and 10 to the corresponding peaks found in the first four fractions, while of an Aroclor 1254 standard. Reynolds most of the interfering compounds found (1970) employed a method similar to ' in wildlife samples were found in the fifth Koeman et al. (1969a) but based his quan zone. Bagley et al. (1970) used this meth-. titation on an average of two or more od but mentioned that zones three and peaks. In addition, his results were re four contained practically all unknown ported as Aroclor 1254 or 1260 depending components as well as p,p'-DDT (zone on the overall pattern of the chroma three) and p,p'-DDE (zone four). Armour tographic peak profile. It is clear that we and Burke (1969) used precoated (alu are still relatively unsophisticated in our minum oxide) sheets and n-heptane and PCB quantitation methodology and will 2% acetone/n-heptane for solvent sys continue to estimate only relative amounts tems. PCBs (Aroclors 1254 and 1260) and of PCBs in field samples until we synthe DDE were not separated, but p,p'-DDT size the individual PCB components com and p,p'-DDD were completely sep monly found in the ecosystem and are arated from PCB by both solvent systems. able to speak in terms of these individual Another possible means of separating in peaks as we do for most pesticide residues. terfering substances is to use a series of However, it has been kindly pointed out differing polarity columns in the gas chro (Risebrough, pers. comm.) that, with ref matograph at the time of determination. erence to biological significance, the cor This is less time consuming and may be rect order of magnitude and accurate rela useful when operating conditions can be tive amounts of PCB give the essential selected such that PCB peaks are absent information. This is because biological in the region where sought pesticides effects, such as enzyme induction, are re emerge (Simmons and Tatton, 1967). lated to degree of chlorination, and the 3)Quantitation. Koeman et al. (1969a) existing methods give some indication of semiquantitatively measured the residues this' activity. Therefore, thatjnformation in Japanese quail fed phenochlor DP6 by would be lost if stress were* placed only using one of the peaks in a phenochlor upon quantitating individual peaks. DP6 mixture as a standard. Risebrough 4) Magnitude of Error. Ever since PCB (1969) quantitated relative levels of PCBs peaks were recognized for what they are, by assuming that each PCB compound residue chemists and other researchers produced the same peak height with the interested in pesticide residues have electron capture detector as the same asked what magnitude of error is likely to amount by weight of p,p'-DDE. After result from ignoring the presence of summing the heights of the individual PCBs. Only recently have studies either peaks, the total was multiplied by a factor directly or indirectly resulted in an esti derived from measurements of standard mate of this error. Anderson et al. (1969) solutions with electron capture and micro- quantitated p,p'-DDE, p,p'-DDD, and coulometric detectors. Jensen et al. (1969) p,p'-DDT in five egg samples before and reported PCB amounts as the sum of all after saponification. There was no appre PCB components and based the estimate ciable change in the p,p'-DDE, but ap on a combination of mass spectrometry parent p,p'-DDD was reduced by ap and microcoulometric and electron cap proximately 58% and p,p'-DDT by 90%. ture detection. Even with this elaborate Reynolds (1970) looked at a large number approach, these investigators suggest that of samples before and after his PCB- the method is still rough and may be cor Florisil separation and found .hat the rect only within a factor of 2. Anderson et actual p,p'-DDD residue (relative to the al. (1969) devised a method for obtaining apparent residue) represented from 0 to a crude estimate of PCB residues as Aro- 7%.in California gulf (Larus occidentalis) dor 1254 from chromatograms where sep fat, 0% in cormorant (Phalacrocorax auri- aration had not been attempted. On an tus) eggs, 80% in 10 pooled mallard (Anas empirical basis, it was found that Aroclor plalyrhynchos) duck eggs, and from 0 to 1254 couid be quantitated by considering 104% in great blue heron (Ardea cinerea) peak 10 as p,p'-DDT and multiplying that eggs. Respective values for p,p'-DDT value by 10. Since this peak had originally were 0 to 53%, 13 to 41%, 100%, and 18 to been quantitated as p,p'-DDT, and in 102%. Actual residue levels of heptachlor epoxide generally represented a majority of the apparent values. Before and after values for p,p'-DDE and dieldrin were not significantly different. Obviously, the magnitude of error may vary depending on the trophic level sampled and certainly with the area from which a sample is col lected (Risebrough et al., 1968, Jensen et al., 1969). Since the publishing of ade quate separation techniques, there is no reason why there should be any error in pesticide quantitation contributed by PCBs. . Toxicology Despite some recent studies, the toxi cology of PCBs remains rather poorly known as compared to that of the chlo rinated hydrocarbon pesticides. For ex ample, no definite work has been done to establish LD50 values for the various formulations of PCBs. 1) Acute, single dose experiments. Tucker and Crabtree (1970) found that a single dose of 100 mg/kg Aroclor 1254 (stomach tubed in oil) was fatal to two out of three rats, while 500 mg/kg was not fatal to three rats. Aroclor 1268 killed one rat out of three at 500 mg/kg, 1000 mg/ kg, 2000 mg/kg, and 4000 mg/kg. Smyth (1931) found that a 4 g/kg (degree of chlo rination not stated) was.nontoxic to guinea pigs and rabbits, but this appears to be due to the fact that material which was given as a paste passed through the intes tine unabsorbed. Tucker and Crabtree (1970) found that Aroclor 1242, 1254, 1260, and 1268 at a dose of 2000 mg/kg was not fatal to mallard ducks (four groups of three birds each). . 2) Acute, feeding experiments. Mon santo Bulletin 306 states that 100 ppm diet had no effect on rats, although no details of the studies were given. The ex periments of Bennett et al. (1938) in which 0.05 g/rat of 65% chlorine biphenyl-was given orally every other day led to 50% mortality. If one assumes a body weight of 200 g, then the alternate day dose is roughly 250 mg/kg, which can be com pared to oral LD50 for DDT of 113 mg/kg (Frear, 1968). Miller (1944) found that two oral doses of 69 mg of 42% chlorine biphenyl a week apart were fatal to guinea pigs. At'an estimated body weight of 400 g, this gives a dose of 170 mg/kg. Tucker and Crabtree (1970) fed rats diets contain ing 10 and 1000 ppm Aroclor 1254. One rat out of six on the low dose died; this mortality was considered to be due to other causes. Four out of four of the high group died within 53 days and the cal culated intake was 1330-1520 mg/kg. The 960 . BioScience Vol. 20 No. 17 DSW 552535 STLCOPCB4090952 food intake of the 1000 ppm group was excess of 2000 ppm: This finding is in hexobarbital, in vitro rates of aniline hy- only 79% of the control group. agreement with the findings of Dale, droxylation and demethylation of p-nitro- Presst et al. (1970) have examined the Stickel, and co-workers that the brain anisole, and the rate of excretion of tokicity of Aroclor 1254 to Bengalese levels are the best indication of acute dieldrin. These workers studied 10 com finches (Lonchura striata). This is a diffi- toxic levels. pounds ranging in chlorine content from cult species for which to calculate the McCune et al. (1962) found no mor 21% to 68% and found that all the effects dietary intake. Due to their dependence tality with chickens fed 100 or 200 ppm increased with increasing chlorine con upon unshelled food, it is only possible to Aroclor 1242 in. their diet for a 4-week tent. For example, 50 ppm of Aroclor present the PCB-laden food for a few period. On diets of 400 ppm and 800 ppm, 1221 reduced hexobarbital sleeping time hours a day (Jefferies, 1967). Loss by the mortalities over a 4-week period were, by 11%, whereas for Aroclor 1248 and evaporation and loss by spillage must be respectively, 50% and 90%. During the 1268 the figures were, respectively, 35% allqwed for, and increase of weight by def first 3 weeks, the mortality figures were and 48%. Thus, the sublethal effects have ecation must be kept to a minimum. 10% and 50%, respectively. Flick et al. direct correlation with chlorine content, Thus, the calculated dietary intake is sub (1965), using the same material at 400 while the lethal, effects appear to be in I ject to more error than is usually the case. ppm in the diet, had three birds out of 24 versely correlated (Tucker and Crabtree, Presst et al. (1970) also measured the con die in a 3-week period. Koeman et al. in press). Lincer and Peakall (1970) noted centration of PCBs in the liver; they (1969a) found that a diet containing 2000 an increase of the in vitro rate of metabo found that the range was large (i.e., 70 ppm Phenoclor DP6 caused complete lism of estradiol in kestrels fed 0.5 and 697 ppm in birds that died compared to mortality with Japanese quail (5/5) in 5 5 ppm Aroclor 1254 in their diet and also 3-634 ppm in those that survived). These 13 days and rats (8/8) in 1-56 days. demonstrated increased levels of cytoplas authors conclude that Aroclor 1254 has Schoettger (unpublished) found that the mic RNA with the higher dietary level only 1/13 the toxicity of DDT, although 96-hr TLm for Aroclor 1221 using cut using a cytophotometric technique. l the different shape of the mortality throat trout was 1.2 mg/1 and for Aroclor Tucker (unpublished) found that a sin curves--steep with DDT, gradual with 1260 was 60.9 mg/1. In general, they gle oral dose of 500 mg/kg Aroclor 1254 PCB--makes comparison difficult. Jef found that the toxicity of Aroclors was caused regular egg laying of Japanese feries and Walker (1966) found good cor inversely proportional to their percentage quail (Coturnix coturnix) to turn first to relation between calculated dietary intake chlorination and directly proportional to scattered egg production and then to stop and liver concentrations for pp'-DDT in their solubilities. These figures suggest completely for a week. The scattered eggs the Bengalese . finch. However, other that PCBs ^re.two to three orders of mag had shells 9% thinner than normal, but workers (Dale et al., 1963; Stickel et al., nitude less toxic to fish than DDT. 'york thickness returned to control values when 1966; Stickel and Stickel, 1969) have con by Lichtenstein et al. (1969) on house and regular laying was resumed. Mallard sidered that levels in the brain are a more fruit flies found that PCBs were 40-300 ducks dosed with 1000 mg/kg of Aroclor reliable index of toxic levels than those in times less toxic than DDT, the toxicity laid one or no eggs before stopping for l- the liver or whole carcass. decreasing as the chlorine content in 2 weeks. The few eggs laid after the sin De Vos and Koeman (1970) fed Pheno- creases. Thus, it appears that the lower gle large-dose of Aroclor had shells 18% cfor DP6, Clophen A60, and Aroclor 1260 vertebrates and invertebrates are much thinner, and again eggshell thickness was to chickens at a dosage of 400 ppm. Mor less susceptible than mammals to direct normal after resumption of regular laying. tality was complete (20/20) for those birds toxicity from PCBs. The period before ejg laying has been on Phenoclor in 12-58 days and in 13-29 3) Sublethal effects. As with the chlo noted to be increased in the ring dove days for Clophen. The mortality for Aro rinated hydrocarbon pesticides, the most (Peakall, unpublished). It is possible that clor was only 3/20 for a 60-day period. important effects are long-range sublethal the mechanism involved is increased rate This differential effect is unexplained and effects. The pathologic changes in various of metabolism of circulating estradiol in is surprising in view of the fact that all organs are summarized in Table 2. The the liver (Peakall, 1970). Anderson et al. three formulations contain 60% chlorine. table .shows some interesting differences (1969) had suggestive evidence that These workers measured the residue lev between mammals and birds. The most PCBs affected eggshell thickness, al els in the liver, and, for some birds, in the striking finds in mammals are alterations though to a less extent than DDE. Pre brain, for chickens dying during the ex to the liver, whereas fluid in the peri liminary results with ring doves support periment. Although there was consider cardial sac, kidney damage, and reduced this conclusion (Peakall, unpublished). able variation, most of the brain levels spleen was found in birds. were between 210 and 420 ppm, which McLaughlin et al. (1963) found that 25 Levels of PCBs Found in Nature can be compared to 50-80 ppm for DDT mg Aroclor 1242 injected into the yolk (Stickel et al., 1966). On the basis of this sac of chicken eggs caused complete mor Roburn (1965), comparing total chlorine work, PCB is 1/4-1/5 as toxic as DDT. Little difference was noted between the three different PCB formulations, al though the numbers of determinations involved was rather small. If this finding tality, whereas 10 mg caused 95% failure and teratogenetic effects were noted among the young that hatched (beak deformity, edema, and growth retardant). Induction of hepatic hydroxylating en (by concentration cell techniques) with results calculated from gas chromatog raphy, found that some unknown chlorine compounds were present in seveial tissue samples and eggs of wild birds in Great is borne out by subsequent work, it would zymes has been demonstrated in the pi Britain. The first identification of these suggest that differential absorption from geon (Risebrough et al., 1968), rat (Street materials was by Jensen and it is stated the gut or differential penetration of the et al., 1969), and American kestrel (Falco (Anonymous, 1966) that residues in feath blood brain barrier is involved. The liver sparverius) (Lincer and Peakall, 1970). ers collected in Sweden go back to 1944. values were more variable. There was a Street and co-workers studied the effects PCB residues have been reported in wild considerable number in the 200-400 ppm of a diet of 50 ppm and 100 ppm on sleep life from Canada (Holden and Marsden, range, but there were several values in ing time induced by a standard dose of 1967; * -J-enn et al., 1969; Reynolds, September 1, 1970 j DSW 552536 961 STLCOPCB4090953 TABLE 2. Pathologic changes induced by PCBs Treatment Animal liver- . ' Kidney Pericatdaiia and Peritoneum Other Observable Changes Rateteaca* Singfi oral doseof 69 mg 142% d) Guinea Pig Rat Rabbit ' Small fat droplets through lobules, slight to moderate central atrophy, focal necrosis noted a few animals. Essentially normal . No noteworthy changes 'i Adrenals, spleen, and pancreas showed no noteworthy changes. MiOai (1544) 300 mg daily for 6 days (65% a) Rat Ceils swollen, hyaline granolas present most died withia few days. -- laaaett Mat (1935) SO mg daily far vp to 6 months iss a) Rat Enlarged (33% weight increase). Urge number of hyaline globdes cytoplasm. Several died doting experiment. Banaatt 25,50, & 100 ppm diet for IS days (21-68% D Arodors) Rat Incteasa in weight, affect increasing with increasing chlorine content Arodor 1232-10%. 1242-12%, 1254 14%. 1258-24% at 50 ppm. Street et af. (Is press) 100 ppm in diet 200 ppm in (Set 400 ppm diet 600 ppm in diet (Arodor 1242) Chicken No effect No effect Enlarged and mottled Damaged . * Damaged Slight Hydroperieerdium Hydropericardtum Hydropericardium, hydroperitoneum. Enlarged." McCuaaatai. (1952) ./ 200 & 400 ppm m (Set for 3 weeks (42%, Arodor) Chicken No changes noted Paleness at 200 ppm, extensive hemorrhage, and enlargement at 400 ppm. Increased fluid in pericardial sac at tha higher concentration. Paleness of pancreas, en largement of adrenal and smaD spleen at low concen trations. At higher concennations pale cream-colored pancreas, adrenals hemorrhagic. Rick itaL (19651 Variout doses (54% a. Arodor) Bengalese Finch No weight changes Weight was 32.4% of brain * weight for controls and 53.5% for those dying from PCB poisoning. Slight weight increase, a few showed liquid * in pericardia! sac. . .' V Breast at al (la press) 400 ppm in (Set for 60 days (6Q% Q-) Chicken Centrolobalar necrosis (compd 1 and 2). Liver weight increased from 2.76 g/100 g to 4.31 g/100 g (compd. 31. Fatty degen . eration. Tubular dilatation, (compd. 1 and 2} Rare with compd. 3. . Hydropericarifium common with compds. 1 and 2. Rare with compd. 3. Increased porphyria, spleen small with reduction of red pulp and atrophy of white pulp (compd. 1 and 2). Spleen decreased from 0.146 g/100 g to 0.136 g/100g. compd. 3). Vos and Koemaa (In press) *n<Mdtf OP 0 (compd. 11.Ooplioa ABO (compd. 2| mid Arodor 1280 (compd. 3) wttt aid. Oifflirontiol afflicts ootod wdor compd. numbon. AS chiekon 4id m compd. 1 and 2 wnhia 80 Wfs.ufy 15% wrtdttyoo compd. 3. 1970), Germany (Fiuczynski and Wendland, 1968; Koeman et al., 1967), Great Britain, (Holmes et al., 1967; Presst and Jefferies, 1969; Presst et al., 1970; Holden and Marsden, 1967), Netherlands (Koe man et al., 1967; 1969), Sweden (Anony mous, 1966; Jensen et al., 1969), and the United States (Anderson et al., 1969; Risebrough et al., 1968; Risebrough, 1969).. Biological Magnification The figures (mean, range of values, and sample size) given-in the table below are taken from their paper. PCBs (ppm in extractable fat) Baltic Stockholm Archipelago Mussel Herring Seal Guillemot eggs 4J (1.9-8.6) (40) 6.8(0.5-23) (18) 34(16-44) ( 3) 250(I40*360)( 9) 5.2 (3.4-7.0) 5.1 (3J-8.5) 30(16-56) -- (15) ( 4) ( 3) White-tailed Eagle No detailed studies, such as those for' DDD at Clear Lake (Hunt and Bischoff, 1960) and DDT and its metabolites in Lake Michigan (Hickey et al., 1966), have yet been made for PCBs. ` Pectoral musde Brain ' Egp Heron -- -- -- -- 14,000 (8400-17,000) ( 4) - 910(490-1500) ( 3) 540(250-800) ( 5) 9400 ( 1) ^ The. most detailed studies currently The levels in three species of fish in Clear available are those of Jensen et al. (1969). Lake in 1968 were 0.03-0.005 ppm (wet weight) compared to 0.098 ppm in the breast muscle of a western-grebe (Rise brough et al., 1969). Anderson et al. (1969) found that in most fish extracts the levels of PCBs were less than 0.1 ppm, whereas the levels in the eggs of cormorants (Phalacrocorax auritus) were 5-9 ppm. Presst, Jefferies, and Moore (unpublished) found that the livers of fish-eating birds in the British Isles ranged up to a maximum of 900 ppm (wet weight), bird feeders up to 70 ppm, mammal eaters to 50 ppm, and insectivores to 1 ppm. Unfortunately, no average values or prey items were in cluded. The physical properties of PCB and the available residue data clearly indicate that these materials are capable of bio logical magnification up the food chain. DSW 552537 962 ' BioScience Vol. 20 No. 17 STLCOPCB4090954 Ratio of DDT to PCB Significance of Current Levels cide residue analysis. F.D.A. Laboratory In Riscbrough et al. (1968) and Risebrough (197.9) have examined the ratio of total In view of the similarity of PCBs to DDT and its metabolites, the addition of formation Bull. No. 918, II p. _______ 1970. A method for separating poly chlorinated biphenyls from DDT and its anal DDT, i.e., DDT and its metabolites to PCB residues to the environment is rough ogs. J. Assoc. OJJic. Anal. Chern. (in press). PCB. He has found that the ratio DDT/ ly equivalent to an increase of DDE resi Bagley, G. E., W. L. Reichel, and E. Cromartie. PCB was 1-2 in San Francisco Bay and 5 10 for seabirds in the Pacific; in the Gulf dues. However, since a synergistic effect of PCBs on DDT has been demonstrated 1970. Identification of polychlorinated bi phenyls in two bald eagles by combined gasliquid chomatography-mass spectrometry. of California, a region relatively remote in insects (Tsao et al., 1953), this- pos J. Assoc. OJJic. Anal. Chem., 53: 251-261. from contamination, the ratio was 9-10. sibility should not be overlooked in higher Bartha, R. 1969. Pesticide residue interaction Vermeer (quoted in Reynolds, 1970) in western Canada found a DDE/PCB ratio of 7 for California gull tissues and 13 for organisms. The enzyme induction effects of PCBs have been well documented, and carbonic anhydrase inhibition is likely. creates hybrid residue. Science. 166: 1299 1300. Bennett, G. A., C. K. Drinker, and M. F. War ren. 1938. Morphological changes in the great blue heron eggs. In the Baltic the The effect of PCBs on photosynthesis is a livers of rats resulting from exposure to cer ratio was 1-2 (Jensen et al., 1969), al critical experiment that has not been tain chlorinated hydrocarbons, J. Ind. Hyg. though along the west coast of Sweden the ratio was as low as 0.15. In grebes in the British Isles the ratio was 0.4-0.8 done. Wurster's (1968) experiments with phytoplankton should be repeated with PCBs. Toxicol.. 20: 97-123. Bourne, W., and C. Mead. 1969. Seabird slaughter. B.T.O. News. No. 36, p. 1-2. Dale, W. E., T. B. Gaines, W. J. Hayes, Jr., and (Presst and Jefferies, 1969). For sea-bird The most critical area for research on G. W. Pearce. 1963. Poisoning by DDT: Re t s t eggs, Presst et al. (unpublished) found ratios of 0.06 to 0.5. The overall impression is that the PCBs is to discover the major source(s) of escape into the environment. Legislative control of a material escaping as a side lation between clinical signs and concentra.tion in rat brain. Science, 142: 1474-1476. Deonier, C. C., H. A. Jones" and H. H. Incha. 1946. Organic compounds effective against amount of PCB in tissues tends to paral effect of its use may have a different set Anopheles quadrimaculatus. Laboratory- lel that of DDE, at least in local ecosys of problems than the control of pesticides test. J. Econ. Entomol.. 39: 459-462. tems, and the DDE/PCB ratio is lowest near industrial areas suggesting that PCB which are broadcast as a function of their normal use. The evidence suggests that it Duda, E. J. 1957. The use of chlorinated poly phenyls to increase the effective insecticidal life of lindane. J. Econ. Entomol.. 50: 218 is not carried quite so readily to remote is important to find the leak and stop it. 219. areas. Nevertheless, the variation of the Erro, F., A. Bevenue, and H. Beckman. 1967. A ratio is small enough to suggest that the method for the determination of toxaphene routes of dispersal are similar. Since the in the presence of DDT. Bull. Environ. Con evidence points to aerial fallout as the * Acknowledgments 4 tam. Toxicol..2: 372-380. Fiuczynski, V. D., and V. Wendland. 1968. The route of dispersal of the chlorinated hy drocarbon pesticides (Risebrough et al., We are grateful to Dr. Koeman (Uni population of Milyus migrans in Berlin 1952 1967. J. Ornilhol.. 109: 462-471. 1968; Frost, 1969), it is likely that this is versity of Utrecht), Drs. Presst and Jef Flick, D. F., R. G.,O'Dell, and V. A. Childs. also the main route for PCBs. The path feries (Nature Conservancy, Great Brit- 1965. Studies of the chick edema disease. 3. ways by which PCBs escape into the eco system are poorly known, although the possibilities have been recently discussed ian). Dr. Reynolds (Ontario Research Foundation), Dr. Street (Utah State Uni versity), and Dr. Tucker (U.S. Fish and Similarity of symptoms produced by feeding chlorinated biphenyl. Poultry Sci., 44: 1460 1465. Frear, D. E. H. 1968. Pesticide Handbook- , at some length (Risebrough, 1970; Rey Wildlife Service) for advance copies of Entoma. College Science Publishers. nolds, 1970). Since a large number of plastics and resins may contain PCBs (Table l), the most likely route is combus important material. Some of the work presented, here was carried out under NIH Grant ES00306, Dr. T. J. Cade, Frost, J. 1969. Earth, air, and water. Environ ment. 11: 15-33. Hickey, J. J., J. A. Keith, and F. B. Coon. 1966. An exploration of pesticides in a Lake Michi tion of these materials. This supposition Principal Investigator. The review was gan ecosystem. J. Appl. Ecol.. 3 (Suppl): remains to be tested. The possibility that written while one of us (D.B.P.) was an 141-154. it PCBs could be derived from DDT should also be considered. The possibility that this conversion occurs in tissue is most Established Investigator, American Heart Association. Thanks go to Dr. T. J. Cade for reviewing the manuscript. Holden, A. V., and K. Marsden. 1967. Organo chlorine residues in seals and porpoises. Nature. 216: 1274-1276. Holmes, D. C., J. H. Simmons, and J. O. 'G. unlikely for two reasons. First, it has never been detected despite the detailed work on the metabolism of DDT. Second, the References Tatton. 1967. Chlorinated hydrocarbons in British wildlife. Nature. 216: 227-229. Homstein, I., and W. N. Sullivan. 1953. The role of chlorinated polyphenyls in improving only mechanism likely to give rise to a lindane residues. /. Econ. Entomol.. 46: 937 i i ( biphenyl is via free radicals and this is unlikely to occur in tissue. However, in Abbott, D. C., R. B. Harrison, J. O'G. Tatton, 940. and J. Thomson. 1966. Organochlorine pesti Hunt, E. G., and A. I. Bischoff. 1960. Inimical the atmosphere under the influence of cides in the atmosphere. Nature. 211: 259 261. effects on wildlife of periodic DDD applica tions to Clear Lake. CaliJ. Fish Came. 46: UV light, such a breakdown is more prob Anderson, D. W,, J. J. Hickey, R. W. Rise 91-106. able. A possible reaction is shown in brough, D. F. Hughes, and R. E. Christen Jefferies, D. J. 1967. The delay in ovulation pro Figure 1. Tautomeric shift could lead to a variety of isomers of dichlorophenyl, . but it is difficult to envision the formation of more highly chlorinated biphenyls by sen. 1969. Significance of chlorinated hydro carbon residues to breeding pelicans and cormorats. Can. Field Natur.. 83: 89-112. Anonymous, 1966. Report of a new chemical hazard. NewSci.. 32: 612. duced by pp'-DDT and its possible signifi cance in the field. Ibis. 109: 266-272. Jefferies, D. J., and C. H. Walker. 1966. Uptake of pp'-DDT and its post-mortem breakdown ia the avian liver. Nature. 212: 533-534. this route. Since PCBs extracted from biological material match well with higher Aroclors (i.e., 1254), it seems unlikely that PCBs found in nature could be derived Archer, T. E., and D. G. Crosby. 1966. Gas chromatographic measurement of to.Xaphene in milk, fat. blood, and alfalfa hay. Bull. Environ. Contam. Toxicol.. I: 70-75. Armour, J., and J. Burke. 1969. Polychlorinated Jensen, S., and G. Widmark. 1967. Organochlo rine residues, OECD preliminary study .1966 67. Report given at the OECD Pesticide Conference. Jensen, S.. A. G. Johnels, S. Olsson, and G. -from other materials. ' biphenyls as potential interference in pesti Otter'' ' "'cn nOT and PCB in marine September I, 1970 DSW 552538 STLCOPCB4090955 - animals from Swedish waters. Nature, 224:' ` 247-250. Klein, A. Ki, and J. O. Watts. 1964. Separation Mulhern, B. 1968. An improved method for the separation and removal of organochlorine insecticides from thin-layer plates. J. Chro- termining organochlorine pesticide residues in wildlife. J. Chromatogr., 27: 253-255. Smyth, H. F. 1931. The toxicity of certain ben and measurement of Perthane, DDD (TDE), matogr.. 34: 556-558. zene derivatives and related compounds. J. and DDT in leafy vegetables by electron Peakall, D. B. 1970. Pesticides and the repro Ind. Hyg. Toxicol., 13: 87-96. capture gas chromatography. J. Assoc. OJJic. duction of birds. Sci.Amer., 222: 73-78. Stickel, L. F., W. H. Stickel, and R. Christensen. Anal. Chem., 47: 311-316. Penning, C. H. 1930. Physical characteristics 1966. Residues of DDT in brains and bodies Koeman, J. H.. A. A. G. Oskamp, J. Veen, E. and commercial possibilities of chlorinated of birds that died on dosage and in survivors. . Brouwer, J. Rooth, P. Zwart, E. V. D. Broek, and H. Van Genderen. 1967. Insecticides as diphenyl. Ind. Eng. Chem., 22: 1180-1182. Presst, I., and D. J. Jefferies. 1969. Winter num Science, 151: 1549-1551. Stickel, L,, and W. Stickel. 1969. Distribution of a factor in the mortality of the Sandwich tern (Sterna sandvicensis). A preliminary communication. Meded. Riijksfac. Land bers, breeding success, and organochlorine residues in the great crested grebe in Britain. Bird Study, 16: 168-185. DDT residues in tissues of birds in relation to mortality, body conditions, and time. Ind. Med. Surg.. 38: 44-53. - bouwwetensch. Gent., 32: 841-853. , Presst, L, D. J. Jefferies, and N. W. Moore. Street, J. C., F. M. Urry, D. J. Wagstaff, and A. Koeman, J. H,, M. C. Ten Noever de Brauw, 1970. Polychlorinated biphenyls in wild birds D. Blau. 1969. Comparative effects of poly and R. H. de Vos. 1969a. Chlorinated bi in Britain and their avian toxicity. Environ. chlorinated biphenyls and organochlorine phenyls in fish, mussels and birds from the Pollut. (in press). pesticides in induction of hepatic microsomal River Rhine and the Netherlands coastal area. Reynolds, L. M. 1969. Polychlorobiphenyls enzyme. Presented at ACS meeting Septem Nature, 221: 1126-1128. (PCBs) and their interference with pesticide ber 8-12, New York. Koeman, J. H., J. A. J. Vink, and J. J. M. de residue analysis. Bull. Environ. Contam. Toxi Sullivan, W. N., and I. Homstein. 1953. Chlo Goeij. 1969b. Causes of mortality in birds of col.. 4: 128-143. rinated polyphenyls to improve lindane resi prey and owls in the Netherlands in the win 1970. Pesticide residue analysis in the dues. J. Econ. Entomol., 46: 158-159. ter of 1968-1969. Ardea. 57: 67-76. presence of polychlorobiphenyls^ (PCBs). Tsao, Ching-Hsi, W. N. Sullivan, and I. Hom Lichtenstein, E. P., K. R. Schulz, T. W. Fuhre- Residue Rev. (in press). stein. 1953. A comparison of evaporation i I mann, andT. T. Liang. 1969. Biological inter action between plasticizers and insecticides. Risebrough, R. W. 1969. Chlorinated hydrocar bons in the global ecosystem. In: Chemical rates and toxicity to house flies of lindane and lindane-chloriated polyphenyl deposits. J. J. Econ. Entomol., 62: 761-765. Fallout, G. G. Berg and M. W. Miller (Eds.), Econ. Entomol., 46: 882-884. Lincer, J. L., and D. B. Peakall. 1970. Induced hepatic steroid metabolism and - increased cytoplasmic RNA by polychlorinated biphe Charles C Thomas, Springfield, 111., p. 5-23. 1970. More letters in the wind. Envi ronment. 12: 16-27. Tucker, R. K., and D. G. Crabtree. 1970. Hand book oj Toxicity o] Pesticides to Wildlife. U.S. Dept, of Interior, Bureau of Sport Fish nyls (PCB) in the American kestrel (Falco Risebrough, R. W., P. Reiche, D. B. Peakall, S. eries and Wildlife, Resources Publication No. 84. sparverius). Nature (in press). McCune, E. L,, J. E. Savage, and B. L. O'Dell. G. Herman, and M. N. Kirven. 1968. Poly chlorinated biphenyls in the global ecosystem. de Vos, J. G., and J. H, Koeman. 1970. Com parative toxicological study with polychlori 1962. Hydropericardium and ascites in chicks Nature, 220: 1098-1102. nated biphenyls in chickens with special ref fed a chlorinated hydrocarbon. Poultry Sci.. Risebrough, R. W., R. Reiche, and H. S. Olcott. erence to'porphyria, edema formation, liver i 41: 295-299. McLaughlin, J., Jr., G. P. Marliac, M. J. Ver- 1969. Current progress in the determination of polychlorinated biphenyls. Bull. Environ. necrosis and tissue residues. Toxicol. Appl. Pharmacol, (in press). { relt, M. K. Mutchier, and O. G. Fritzlaugh. Contam. Toxicol., 4: 192-201. Widmark, G. 1967. Possible interference by 1963. The injection of chemicals into the yolk Roburn, J. 1965. A simple concentration-cell chlorinated biphenyls, J. Assoc. OJJic. Anal. sac of fertile eggs prior to incubation as technique for determining small amounts of Chem., 50: 1069. . toxicity test. Toxicol. Appl. Pharmacol. 5: halide ions and its use in.the-determination 1968. Determination of number of com 760-771. of residues of organochlorine pesticides. pounds which can result from the chlorination Miller, J. W. 1944. Pathologic changes in ani Analyst, 90: 467-475. ' of biphenyl, and development of a simple sys mals exposed to a commercial chlorinated Schmidt, H., and G. Shultz. 1881. Einwirkung tem by which these may be codified. OECD diphenyl. U.S. Pub. Health Rec.. 59: 1085 von Fiinffach-Chlorphosphor auf das y- report Sweden. 1093. diphenol. Ann. Chem.. 207: 338-344. Wurster, C. F. 1968. DDT reduces photosynthe Monsanto Technical Bulletins 0/PL-311A, Simmons, J. H., and J. O'G Talton. 1967. Im sis by marine phytoplankton. Science. 159: O/PL-306, and O-FF/1. proved gas chromatographic systems for de 1474-1475. MICROBIAL CULTURES COMPLETE BSCS LISTING! GUARANTEED PURITY1 OVER 200 SPECIES AVAILABLE I MOST ORDERS FILLED WITHIN 48 HOURS NOTICE l --------0 BACTERIA AND FUNGI CULTURES-------- 53,00 PER CULTURE - ($2.50 per culture for orders of 6 or more cultures) C s-- BACTERIOPHAGE ---- $4.00 per 5 ml filtered broth suspensions C m-----0 BACTERIOPHAGE CULTURE SETS---------- Available at special prices . COMPLIMENTARY CULTURES One complimentary culture (bacterial or fungal) will be included for each six cultures ordered. Compliments may be selected if so desired. Free CATALOGUE OF CULTURES Available Upon Request DSW 552539 0PRESQUE ISLE MICROBIOLOGICAL ASSOCIATES P.O.BOX 8007*PRESQUE ISLE, PENNSYLVANIA 16505 Telephone 814-833-6373 STLCOPCB4090956