Document 7OqrRM986KvmEqa1L0JMVOVe6

PCB LEVELS IN EGG YOLKS ASSOCIATED WITH EMBRYONIC MORTALITY AND DEFORMITY OF HATCHED CHICKS Casimir Francis Tlmasonis, Brian Bcsji. and Frederick Donald Baker Division of Laboratories and Research. ' Aew York State Department of Health Albany, A'. Y. 12201 The effect of exposing White Leghorn hens for six weeks to 50 ppm of Aroclor 1254 in water was investigated. While egg production decreased temporarily, no significant effects were noted on egg weights or fertility. Incubation of eggs from exposed hens indicated that this polychlorinated biphenyl (I'CB) did not interfere with gametogenesis and fertilization. As Aroclor 1254 accumulated in the yolk, however, embryonic development was arrested at progressively earlier develop mental stages. When I'CB was removed from the water there was an initial rapid decline in the PCB content of the yolk and a slow return of the normal develop mental pattern of the embryo. Leg, toe. and neck deformities were present in many of the chicks hatched from eggs in which yolk PCB level was 10-15 ppm or more. The gjoss appearance of the liver and the presence of hemorrhages suggest that the liver and the vascular elements may be targets for PCB action. . Introduction The widespread distribution of polychlorinated biphenyls (PCB) in the environment has become a matter of growing concern (Peakall and Linccr l.l>70. Risebrough ct at. 1968). This concern is based, not only on the apparent involvement of PCB in the die-off of some fish and waterfowl, the abnormalities noted in young terns from Long Island Sound (Hays and Risebrough 1972), and the reproductive failure of other avian wildlife species, but also on its cumulative character as one proceeds up the food chain to inan. Sufficient evidence exists to indict PCB as cnibryotoxic in rabbits (Grant ct al. 1971) and to implicate it in chromosomal alterations (Peakall ct at. 1972), metabolic derangements (Bailey and Bunyan 1972, Platonow and Funncll 1972. Pardiui 1971, Vos <7 al. 1971), and dermatological disorders (Vos and Beems 1971) of experimental animals. Yuslto, a poisoning caused by PCB contamination of rice oil, has established its toxicity to humans (Kuralsune ct at. 1971). For an extensive review of PCB toxicity, see Fishbein (1972). Studies of the acute and sublethal toxicity levels of PCB have been done oil a number of avian species and have established the rather high dosage requiied to kill birds (Koeman ct al. 1969, McCune ct al. |9o2, and McLaughlin cf al. 1963). Peakall (1971) has shown Archives nf KnvironnirnLil Contamination and Toxicology Vol. I, No. 4, 107.1, <) J07J by Springer -- Vcrlap New York'lnc. DSW 363889 STLCOPCB4098456 mrifc . . ITA LITY HICKS Donald Raker f i i I f I : \ i lo 50 ppm of Aroclor ased temporarily, no ubation of eggs from 2B) did not interfere mulatcd in the yolk, vely earlier devclop; was an initial rapid the normal developwere present in many 10-15 ppm or more, rages suggest that the f 1 in the environment 0, Risebrough et al. Df PCB in the die-off ns from Long Island ` othc-r avian wildlife food chain to man. rant dal. 1971) and abolic derangements 1. Vos ct al. 1971), al animals. Yusho, a s toxicity to humans xe Kishbein (1972). n done on a number 3 kill birds (Kucman ill (1971) has shown PCB in Egg Yolks and Embryonic Mortality of Chicks 313 that egg shell thinning did not occur in ringdoves. Previously, Peakall and Linccr(1970) cited Tucker's unpublished data that following PCB administration to Japanese quail and mallards, not only egg shell thinning but.also a temporary cessation of egg laying occurred. A depressed growth rate of newly hatched chicks was noted with PCB levels of 50-150 ppm in feed (Rehfcld cl al. 1971). The PCB content in the eggs of pelicans and cormorants has been reported (Anderson cl al. I960); in the latter case, there is some evidence link ing the increase to the birds' feeding habits (Zitk'o and Choi 1972). In pheasants, while fertility and hatchability of eggs, mortality of adults, and eggshell thickness were apparent ly not affected by PCB. the total reproductive success was depressed (Dahlgren and Linder 1971). Peakall et al. (1972) have found that exposure to 10 ppm of Aroclor 1254 for three months had no effect on the development of eggs of ringdoves; however, six months later embryonic mortality was nearly 100 percent. Scott cl al (1971) found a significant decrease in the hatchability of chicken eggs after treatment for eight weeks with 10 ppm of Aroclor 1254; treatment with 20 ppm caused complete failure, with mortality occur ring immediately before hatching. This report presents preliminary data on the effects of six weeks' exposure to 50 ppm of Aroclor 1254 in water upon the egg production of mature White Leghorn hens, the PCB levels in the yolks, the fertility of the eggs, the viability and hatchability of the embryos, and the deformities present in the hatched chicks. Data arc also presented for the following 20-week clearance period on PCB content of the yolks and the viability and hatching percentages of the embryos. Materials and methods Twenty-four White Leghorn hens, five months old, selected as active layers, were legbanded and randomly segregated into a control and an experimental dock. Each flock initially had 12 hens plus one rooster. Water and breeder crumbles (egg-laying mash) were supplied ad lib. The egg production of each group was between 70 and 75 percent. " Experimental hens were placed on a water ration containing 50 ppm of Aroclor 1254 (Monsanto. St. Louis, Mo. 63121). The water ration was prepared by emulsifying 0.56 g of Aroclor 1254 in 15.0 g of Tween SO (polyoxyethylene sorbitan mono-oleate: Sigma Chemical Co., St. Louis. Mo. 631 IS) at 60 C for ouc-half hour using the Pyro-multimagnestir (Lab-Line Instruments, Inc., Melrose Park, III. 60160). Subsequently, 100 ml of glass-distilled water was added and the mixing continued for another one-half hour. The resultant emulsion was then stirred into 11.5 liters of water prior to being placed in the water dispenser. The average daily intake of water by both the control and experimental hens was recorded over the six-week exposure period and for two weeks thereafter. Egg pro duction was recorded daily, and the eggs were weighed. During the first four weeks, four eggs were randomly selected from those collected daily from each flock, hard-boiled, and stored in a freezer for PCB analysis. After the fourth week this was done weekly. DSW 363890 STLCOPCB4098457 314 C. F. Tumasonis ct at. Eggs not reserved for PCB analysis were placed in an incubator-hatcher and candled on the fifth, twelfth, and seventeenth day of incubation. Nonfertile eggs were discarded. When candling showed the presence of a dead embryo, the egg was opened, from the de gree of embryonic development plus the knowledge of the day the egg was set in the incubator, a reasonable estimate of the age of the dead embryo was possible. Fertility, viability, and hatching percentages were determined for each week's batch of experimental eggs. Equivalent data on control eggs were obtained during the first four weeks of the Study (135 control eggs) and weeks 15-l.S of the clearance period (158 control eggs). Percentages of nonviable embryos and hatched chicks w-crc based on the number of fertile eggs in each week's set. While no experimental hens were sacrificed for analysis of their tissues, spot checks of the blood of four hens on days 11 and 24 on PCB confirmed that it was being picked up. This was no longer considered necessary when yolks continued to show increasing PCB levels. After six weeks, when the hens were returned to the control diet, the rate and the degree of PCB clearance were determined hy analysis of the yolk. Viability and hatchability percentages were recorded to determine the relationship between PCB levels of the yolk and its interference with normal development. One experimental hen died during week eight off PCB and a second during week 13 off PCB. Both were autopsied, and their brains, livers, and muscle tissues were analyzed for PCB residues. To compare the PCB levels in the yolks with those in the hatched chicks, analyses of the liver, brain, and breast muscle were projected. However, in week three on PCB, hatching in the experimental Hock ceased and did not resume until several weeks off PCB. Data on PCB levels in hatched chicks will be presented in a subsequent paper. ' V ' PCB was extracted from yolks and tissues by the method of Richardson et al. (1971). Yolks were used instead of whole eggs because of the lipophilic nature of PCB. Assay was by thin-layer chromatography. Cl500 Silica Gel acid-fast plates (Shlcicltcr and Schucll, Inc., Keene, N.H. 03431) were spotted with samples and standards and were developed in Nanogradc hexane (Mullinckrodi). Subsequently plates were sprayed with silver nitrate dissolved in water and glycerol and then irradiated with ultraviolet for 15 min. Spot densities were determined on a V1S-UV Chromatogram Analyzer (Parrand Optica! Co., Inc., Mt. Vernon, N.Y. 11151) using the wide slit. Details of the modified method for PCB analysis of egg yolks and tissues have been published elsewhere (Bush and Lo 1972). Results I. Weight of hens. At the start of the study, the average weight of the hens of control and experimental tlv>cks was 3.0 and 3.7 lb respectively. Atlci six weeks'exposure there was virtually no change. At the termination ot the study, after some fluctuations, con trols averaged 3.5 lb and experimental hens, 3.4 lb. Slight weight loss had occurred in six of the ten surviving control liens and in nine of the ten surviving experimental hens. --r- DSW 363891 STLCOPCB4098458 itchct and candied on eggs were discarded. opened. From the dche egg was se( in the vas possible. Fertility, batch of experimental irst four weeks ol the d (158 control eggs). t the number of fertile tissues, spot checks of t was being picked up. v show increasing PCB >1 diet, the rate and the t. Viability and hatchbetween PCB levels of ucntal hen died during re autopsied. and their chicks, analyses of the three on PCB, hatching .1 weeks off PCB. Data aper. ichardson ct al. (1971). rure of PCB. Assay was (Shleicher and Schucll, > and were developed in ayed with silver niirate iolet for 15 min. Spot r (Farrand Optical Co., >dificd method for PCB (Bush and Lo 1972). 1 } | ' : , i j : ' ` . . ( | =_ j i :- ; ; , :t of the liens of control x weeks' exposure there some fluctuations, conloss had occurred in six ring experimental hens. ; ; j j i PCB in Fgg Yolks and Embryonic Mortality of Chicks 315 2. Water consumption. Average daily water consumption per hen over the first two months of the experiment was 212 ml for the control group and 177 ml for the experi mental hens. While this difference was present from the start of the experiment, it did not affect egg weights. (Average egg weights during this period were 52.8 g for the controls and 53.5 g for the experimental Hock). 3. PCB levels in dead experimental hens. The two experimental liens that died during the course of this study had been off PCB for more than seven and twelve weeks respec tively. Their livers, brains, and muscle tissues were analyzed for PCB residues, and the levels obtained showed a decline, less in the liver titan elsewhere. The levels were (in ppm): liver, 56.2, 42.5: brain, 52.7, 20.0: muscle tissue, 12.0.3.2. It is as yet unknown, whether these arc toxic levels in the White Leghorn hen. In three hens sacrificed on the conclusion of the study, PCB residues averaged 2.9 ppm in the liver and less than 0.5 ppm in brain and muscle tissue. 4. Egg production and weight. The average daily production of the control (lock was 8.6 eggs during the first six weeks and 7.3 eggs in the subsequent twenty weeks. This de cline reflected three cases of bumblefoot, as well as the normal decline in egg production occurring during warm weather. The average number of eggs collected daily from the experimental flock for each week of the study is shown in Table I. Two experimental hens died, the first during week seven off PCB, the second during week 13 off PCB. While no definitive cause could be assigned for these deaths, autopsy findings indicated that the hens had been unproductive for an undetermined time. The loss of these hens and the influence of warm weather, however, arc not sufficient to explain the temporary decline in egg production of the experimental flock during weeks 3 to 1 2 off PCB. The average weight of eggs from the control and experimental hens over the 26 weeks was 53.8 and 52.9 g respectively. No apparent thinness or fragility of experimental eggs was observed. 5. Egg fertility. The percentage of egg fertility of (he experimental flock for each week of the study is given in Table I. The consistently high percentages indicate that the PCB does not interfere with fertilization. Fertility of control eggs during the first four weeks of the study averaged 92.3 percent; during weeks 15 to 18 off PCB. 93.0 percent. 6. Nonviahility of chick embryos by week of incubation. The percentages of nonviablc embryos in incubated eggs from experimental liens are given in Table I.The distribution of nonviablc control embryos for each of the three weeks of incubation during tire first four weeks of the study was 4.6, 0.5, and 10.1 percent. During weeks 15 to 18 olf PCB, these control percentages were 5.2, 0.6, and 8.3 percent. DSW 363892 STLCOPCB4098459 1-. ' . 316 ' C. F. Tumasonis ct al. Table I. Effect of PCD on c.qy production, fertility, hatching, and embryonic viability of White Leghorn hens* Week Average daily egg production of flockb Fertility percentage Nonviahle embryo percentage Hatching by incubation percentage1' ' 1 week 2 week 3 week During 6 wk on 50 ppm Aroclor 1254 1 9.9 2 9.3 3 7.7 4 9.0 5 8.4 6 8.3 98 0 100.0 100.0 95.0 95.0 95.0 82.7 34.4 0 0 0 0 8.6 1.3 0 11.2 43.2 98.2 8.7 0 9.1 40.0 56.8 1.8 0 64.3 90.9 48.8 0 0 During 20 wk following the exposure period I 8.3 2 8.4 3 6.4 4 8.1 5 7.1 6 6.1 7 4.4 8 4.9 9 6.3 10 5.2 11 5.6 12 5.9 13 6.0f 14 7.1 IS 6.3 16 7.3 17 7.0 18 6.9 19 . 7.0 20 7.0 85.7 95.0 100.0 100.0 94.0 100.0 90.0 91.6 100.0 91.6 100.0 100.0 8.1.2 94.1 85.7 94.1 90.8 100.0 .94.4 95.0 0 0 0 0 11.84 0 0 0 6.2 9.1 26.6 46.1 61.5 75.0 66.6 77.7 85.0 84.6 82.5 89.4 100.0 94.5 24.0 7.0 29.4 33.3 22.0 0 0 0 0 . 7.7 7.7 6.25 8.3 0 5.0 7.7 11.7 0 0 5.5 76.0 93.0 47.1 66.6 33.3 54.5 50.0 72.7 20.0 15.4 7.7 6.25 8.3 0 0 7.7 5.8 0 0 0 0 0 11.7 0 44.5 45.5 43.8 18.1 53.3 . 30.7 23.1 0 16.6 22.3 . 10.0 O' 5.3 5.3 Control values are given in the text under the appropriate headings. bPcr group of twelve hens initially, redueed by deaths in weeks 7 and I 3 off I'C'H. cllatching percentages do not include clucks that "pipped" hut died before hatching. `Hatching not normal; shells were removed to assist chick. *One experimental hen died Indore completion of the seventh week olf I'CIJ. ^Second experimental hen died before completion of the thirteenth week olf I'CU. DSW 363893 STLCOPCB4098460 ^ i jy . Vr. o percentage ation k 3 week 0 64.3 90.9 48.8 0 0 PCB in Egg Yolks and Embryonic Mortality of Chicks - 317 In incubated eggs from experimental liens, complete mortality of first-week embryos occurred during the first week off ECU (Fig. 1). For chick embryos aged 8 to 14 days of incubation, peaks of high mortality were observed in eggs obtained during weeks 4 and 5 on PCB and again during weeks 3 to 12 off PCB. Third-week embryos also exhibited two peaks of high mortality: in eggs obtained from weeks 2 to 4 on PCB and weeks 7 to 16 off PCB. With the removal of Arochlor-I 254-contaminated water from the experimental flock, a withdrawal effect was observed in the percentage and time span that nonviable chick embryos aged S to 14 and 15 to 21 days were found. ' 7. Hatchability. The hatching percentages of experimental eggs arc given in Table I and arc correlated with yolk PCB levels in Fig. 2. For control hens, the average hatching per ccntage was 84.7 during weeks 1 to 4 on PCB and 83.9 during weeks 15 to 18 off PCB. 0 0 0 0 11.7 0 44.5 45.5 43.8 18.1 53.3 30.7 23.1 0 16.6 22.3 10.0 0 5.3 5.3* I- PCB. hatching. 1. f PCB. i i ! i t !I| ( Week ol set Fig. I. Total percentage of nonviable embryos for each week's set. Each bar represents the cumulative percentage of dead embryos of incubation ages, one, two, and/or three weeks found in that set. DSW 363894 STLCOPCB4098461 :. . i 'i 1 318. '. C. F. Tumasonis ct al. Hatching of experimental eggs dropped sharply during the second week on PCB. During . the next eight weeks, no chicks were hatched. During the filth week off PCB two chicks were hatched and were assisted by having their shells removed. Four weeks later, normal hatching of chicks resumed, but the number remained low. The percentage increased pro- gressivcly. however, and by week 17 off PCB it had returned to normal levels. 8. PCB levels in yolks. Fig. 2 shows the PCB levels in yolks of eggs from experimental hens over the 26-week span. During the exposure period these levels increased rapidly. . This suggests not only that PCB accumulates within'thc hen but also that the egg can serve as an excretory mechanism for PCB (Cecil ct al. 1672. Dalilgren ct al. 167 I). These levels also show a close correlation with embiyo nonviability. Moreover, as the PCB level dropped, mortality of the nonviablc embryos occurred at a later time during the incubation period (Fig. i). ` It is apparent that PCB clearance is a protracted process (see Fig. 2). Wien Aroclor 1254 was removed from the water ration of the experimental hens, the yolk levels of PCB '- -. Week of SCI Fig. 2. Relationship between the hatchahility of embryos of each week's set of eggs from experimental hens and the average Aroclor I 254 level in yolks of that set. DSW 363895 STLCOPCB4098462 ek on PCB. During, >ff PCB two ducks vccks later, normal tape increased pro levels. __ 'ront experimental increased rapidly. t the egg can serve 971). Tliesc levels PCB level dropped. incubation period 2). When Arodor yolk levels of PCB i I i i i V ? ' >r r ` ' . 1 *. i\ i i I11 .t..-;y.Aj>.y- s ri` fr-itifffr tiki j,iiiih i iui._ PCB in Egg Yolks and Embryonic Mortality of Chicks 319 dropped quickly for five tveeks and then more gradually over the remainder of the experi mental period. After 20 weeks off PCB. residues in yolk were still present, but only at 0.7 ppm. Since normal hatching percentages were obtained in sets beginning 17 weeks off PCB, it scents apparent that yolk levels under 10-15 ppm have no adverse influence upon cluck embryo development. 9. Condition of chicks hatched front eggs of experimental hens. Chicks hatched from eggs laid by the experimental hens during the first week of exposure were normal. In the set laid the end of the second week along with the drop in the hatching percentage, a de formity characterized by short, bowed legs and crooked and/or clenched toes appeared in two chicks. During the next 12 weeks (three to six on PCB* one to eight off PCB) only two chicks were hatched, both from the set laid in the fifth week off PCB. Both chicks showed this deformity. Chicks hatching from eggs laid during weeks 9 to 13 off PCB . began to show a moderation in the decree of leg and toe deformity and in the percentage of chicks affected. In sets front weeks 14 to 18, toe deformities were usually found only on one foot. ' Abnormal curvature of the neck and head position were present in several chicks hatclung front eggs laid during weeks 11 to 16 off PCB (Fig. 3). In such chicks from sets set of eggs from Fig. 3. Toe and neck deformities present in one-day-old chicks hatched from eggs bid by White Leghorn hens I (i weeks off PCB. The incurving of the toes is typical of many of the chicks hatched from eggs of experimental hens during the course of the study. i DSW 363896 STLCOPCB4098463 320 C. F. Tumasonis ct al. Jaid during weeks 11 and 1 2. this condition interfered with eating and drinking activity, and the chicks died within a week. In chicks from sets laid during weeks 13 to l(>, the condi tion lasted only two to three days before a near-normal head and nock position was attained. These chicks have survived. This condition was not present in chicks from later sets, although several with deformed toes on one foot were noted. Control chicks showed no anatomical defects. Other external conditions noted in chicks hatched from eggs laid during weeks 5 and 9 to 12 off PCB were small size; limited walking activity with absence of flight or scatter reaction; difficulty in breathing, deglutition, and eating; irregular, tufty, and dark dowrv feathcrs;and body tremors. These were not noted in all chicks and were not evident in any chick hatched from eggs laid during or after the thirteenth week off PCB. On autopsy or sacrifice of chicks hatched from eggs laid during weeks'5 to 12 off PCB. the following conditions were noted; extreme hypotrophy of breast muscle tissue; en largement of the gall bladder, enlargement and an abnormal brownish yellow color of the liver, small areas of hemorrhage in the muscle tissue, clotted blood in the abdominal cavity, congested cloacal area, and- resorbed yolk sac engorged with mucoid material. These anomalies were not always present, and with the exception of the pale color of the liv.er, they were not found in any chicks from subsequent sets. Many of the conditions noted are in substantial agreement with those found by Rehfcld et at. (1972) in one-dayold chicks fed Aroclor 1 248. ` Discussion While embryonic deaths do occur during the incubation of normal eggs, the greatly increased percentages of nonviable embryos in eggs from exposed liens during each week of development demonstrate the lethal effect of PCB. The data also show that as PCB levels in the yolk increase, embryonic development terminates at progressively earlier stages. All embryos, however, had developed for at least 48 to 72 hours of incubation, even in sets where mortality of first-week embryos was 100 percent. At this stage, the yolk sac and vitelline circulation have developed sufficiently to begin the transport of deutoplasmic materials to the embryo. The close correlation of the PCB content of the yolks with the nonviability of embryos from these eggs emphasizes the effect of PCB. One unexplained result of this experiment is the absence of nonviabilily in the first-week embryos of sets laiJ during the weeks 8 to I I off PCB. While the same anomaly in three other isolated weeks (week 3 on PCB. weeks 16 and 20 off PCB) falls within the possibilities of experimental, variation, this suslaineJ phenomenon remains hafiling. The manner in which IVU interferes with the developmental ptoccss is as yet unknown. Pardini (1971), using beef hearts, has demonstrated in vitro that PCB inhibits the milochondtial electron transport system on the substrate side of cytochrome C, i.e., NAD11- DSW 363897 STLCOPCB4098464 _____^d'lHi--------r~^-r- r n-`lniriiv---------- i - I t y- - i r t Tit rr! --- ' it iium'Ii i> VimVi <n ri i li-'V'iW* i h *' ni it' i_*j drinking activity, i(o 16, the conditcck position was i chicks from later rol chicks showed uring weeks 5 and of flight or scatter y, and dark down not evident in any L s 5 to 12 off PCD, muscle tissue; cnh yellow color of > in the abdominal mucoid material, e pale color of the of the conditions 1972) in one-day- l eggs, the greatly during each week show that as PCB ogressively earlier f incubation, even itage, tire yolk sac t of deutoplasmic the yolks with the mviability in the the same anomaly B) falls within the remains bafiling. s as yet unknown, inhibits the mitoie C, i.c., NADII- PCB in Fgg Yolks and Embryonic Mortality of Chicks 321 cytochrome C reductase and succinatc-cytochromc C reductase. Peakall ct at. (1972) sug gests that chromosomal alterations caused by Aroclor 1254 can terminate embryonic development in ringdoves. Some of the toxic actions of PCB noted in chickens (Plick cl at. 1965, McCunc ct at. 1962, and Vos and Kocman 1970) may also affect tissue and organ anlagcn during embryogencsis. McCunc c( at. (1962) have shown that four weeks' exposure "to 100 and 200 ppm PCB caused no mortality in chickens. Flick cl at. (1965) have repotted that 3 out of 24 chicks died during three weeks' exposure to 400 ppm Aroclor 1242. Rchleld ct at. (1972) have concluded from their studies of the effects of Aroclor 124S that the maximum tolerance of one-day-old chicks is approximately 50 ppm. McLaughlin ct at. (1963) have shown that 25 mg of Aroclor 1242 injected into the yolk sac of chicken eggs resulted in complete mortality; 10 mg resulted in 95 percent failure to develop, and in teratogenic effects. Cecil ct at. (1972) have studied the effects of a number of PCBs in laying hens. With 2 and 20 ppm of Aroclor 1254 in feed for eight weeks, egg weights and fertility remained un changed, but hatchability and egg production declined. Reduction in halchability was greater with 20 ppm of Aroclor 1232, 1242. and 1248 in teed than with 20 ppm of Aroclor 1254. These studies suggest that the toxicity of PCB is age-, dose-, and type-re lated. While PCB can be retained and accumulated in the body with chronic exposure (Berlin 1970), their elimination does occur mctabolically (Bagley et at. 1970. Cecil ct at. 1972, Kocman et at. 1969, Rehfcld ct at. 1972, Reynolds 1969) as well as by excretion (Cecil ct at. 1972, Dahlgren ct at. 1971). Figure 2 and data on PCB levels in aulopsied and sacrificed hens suggest that effective PCB clearance can be attained. Although chromato graphic studies demonstrate that PCB is metabolizable (Bagley ct at. 1970, Cecil ct at. 1972, Kocman ct at. 1969, Rehfcld ct at. 1971. and Reynolds, 1969), only minor changes in Aroclor 1254 take place in the cockerel (Plalonow and Funncll 1971). Preliminary analytical studies point to sex-related differences in the metabolism of Aroclor 1254 in White Leghorn liens and roosters (Bush and Dell'Acqua 1972). Our observations on chicks hatched from Cggs kid during the weeks off PCB arc in substantial agreement with several of those by Rehfcld ct at. (1971) on one-day-old thicks on an Aroclor 124S regimen. Respiratory and swallowing difficulties were in evidence. Small discrete areas of hemorrhage were noted in muscle tissue, and dotted blood in the abdominal cavity suggested the involvement of PCB in vessel degeneration. Enlargement and discoloration of the liver indicated an effect of IX-'B on hepatic (unction. In addition, we have noted deformities in the legs and toes and an abnormal head position and curvature of the neck. Exposure of White Leghorn hens and roosters to 50 ppm of Aroclor 1254 in water for six weeks did not affect their ability to produce functional gametes, nor did it repress sex ual activity. The failure of the embryos to develop and to hatch, as well as the deformities " . i? T T 4 mji DSW 363898 STLCOPCB4098465 322 C. F. Tuniasonis ct al. present in hatched chicks. is attributable to the accumulated levels of I'd! in the yolk. However, levels found in the yolks after 17 weeks had no adverse effect on embryonic viability. This evidence raises the possibility that excretion of PCB via the egg is a signifi cant cause of the population decline of several species of waterfowl and avian wildlife. Acknowledgments The authors arc grateful to Dr. Melvin K. Abclselh for his discussion and critical review. The excellent technical assistance of Richard Langcnbaeh. Janis Barron. Diane Gould, and John Snow is gratefully acknowledged. References Anderson. D. \V,, J. J. Hickey. R. \V. Risebrough. D. F. Hughes, and R. H. Christensen: Significance of chlorinated hydrocarbon residues to breeding pelicans and cormor ants. Can. Field-Naturalist 83.91 (1969). Baglcy, G. E., W. L. Reichcl. and E. Cromartic: Identification of polychlorinated biphenyls in two bald eagles by combined gas-liquid chromatography-mass spectro metry. J. Assoc. Offic. Anal. Chem. S3, 25 I ( 1970). Bailey, S., and P. J. Bunyan: Interpretation of persistence and effects of polychlorinated biphenyls in birds. Nature 236, 34 (1972). Berlin, M.: I'CB effects on mammals. National Swedish Environ. Protection Board 43-50 PCB Conf.. Sept. 29, 1070, Solna. Sweden. Bush, B.. and R. Dell'Aequa: Personal communication (1072). Bush, B.. and F. C. Lo: Thin-layer chromatography for quantitative polychlorinated biphenyl analysis. J. Chrbmatog. 7, 377 ( 1973). Cecil, H. C., J. Bitman, G. F. Fries, L. 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