1090 King Georges Post Rd. Suite 407, Edison, NJ 08837 908-225-6266 TtCHMCAL ASSISTANCE TEAM FOR EMERGENCY RESPONSE REMOVALAND PREVENTION EPA CONTRACT 68-VVO-0036 21 January 1993 Eileen Helmer U.S. EPA Mail Code 5HSM-TUB7 230 South Dearborn Street Chicago, IL 60604-1602 Dear Eileen, Mark Sprenger requested that I send you the following information: • The exposure model for mink that was used in the NL risk assessment • Life history information for red-winged blackbirds • Data on dietary toxicity of PCBs to mink Enclosed are summary tables with the above information, and a list of references. The soil ingestion rate that was used for mink was obtained from the Wildlife Exposure Factors Handbook, which has only been released as a draft and is not a citeable source. We have tried to obtain a copy of the original study, but have not been successful. We do not have a readily available reference for soil ingestion by passerine birds, but I will try to find one at the library. Also enclosed is a copy of the Cannelton XRF report. This may help indicate what is needed for the Circle Smelting XRF report. Specific information that is needed includes discussion of methods used for sample prep and analysis, the model used for the XMET, and detection and quantitation limits. If you have any questions, please contact me. Sincerely, Roy F. Western, Inc. MAJOR PROGRAMS DIVISION „„ , ._ .„,,.. In Association with Foster Wheeler Enviresponse, Inc., Resource Applications, Inc., C.C. Johnson & Malhotra, R.E. Sarriera Associates, and GRB Environmental Services, Inc.
Transcript
1090 King Georges Post Rd.Suite 407, Edison, NJ 08837 908-225-6266
TtCHMCAL ASSISTANCE TEAM FOR EMERGENCY RESPONSE REMOVAL AND PREVENTIONEPA CONTRACT 68-VVO-0036
21 January 1993
Eileen HelmerU.S. EPAMail Code 5HSM-TUB7230 South Dearborn StreetChicago, IL 60604-1602
Dear Eileen,
Mark Sprenger requested that I send you the following information:• The exposure model for mink that was used in the NL risk assessment• Life history information for red-winged blackbirds• Data on dietary toxicity of PCBs to mink
Enclosed are summary tables with the above information, and a list of references. The soilingestion rate that was used for mink was obtained from the Wildlife Exposure FactorsHandbook, which has only been released as a draft and is not a citeable source. We have triedto obtain a copy of the original study, but have not been successful. We do not have areadily available reference for soil ingestion by passerine birds, but I will try to find one at thelibrary.
Also enclosed is a copy of the Cannelton XRF report. This may help indicate what is neededfor the Circle Smelting XRF report. Specific information that is needed includes discussion ofmethods used for sample prep and analysis, the model used for the XMET, and detection andquantitation limits.
If you have any questions, please contact me.
Sincerely,
Roy F. Western, Inc.MAJOR PROGRAMS DIVISION „„ , ._ . „ , , . .In Association with Foster Wheeler Enviresponse, Inc., Resource Applications, Inc., C.C. Johnson & Malhotra,R.E. Sarriera Associates, and GRB Environmental Services, Inc.
TABLE?
LIFE HISTORY INFORMATION USED IN DIETARY EXPOSURE CALCULATIONSFOR MINK (Mustela vison)
Body Weight: 1 kg (Bleavins et al. 1980)
Dietary Ingestion Rate: 150 g/day (Bleavins et al. 1980)
Home Range: 2630 m stream length, d1850 m stream length, 9 (Linscombe et al. 1982)
Soil Ingestion Rate: 4.2 g/day" (Beyer et al. 1991)
Water Ingestion Rate: 0.10 L/day (Calder and Braun 1983)
Diet: 50% Small Mammals50% Aquatic Biota (Frogs and/or Fish)
"Estimated soil ingestion rate (2.8% of dietary ingestion rate) using red fox data, soil ingestionrates for mink are not available.
TABLE 3. ASSUMPTIONS FOR DAILY INTAKE CALCULATIONS FOR RED-WINGEDBLACKBIRD (Agelaius phoeniceus)
Body weight:
Ingestion rate:
Diet:
Home range:
Soil ingestion rate:
Male, 63.6 gFemale, 41.5 g (Clench and Leberman 1978)
8.4 g/day (Kenaga 1973)*
Primarily aquatic insects (observations made during breeding season;Orians 1980)
73% vegetable matter, 26.6% animal matter (based on year-roundobservations; feed mainly in marsh during breeding season; Bent1965)
Food delivered to nestlings: 97.8% animal matter (by volume;Snelling 1968)
3000 m2 (Nero 1956)
689 m2 in marshes, 2188 m2 in upland areas (Case and Hewirt 1963)
Water ingestion rate: 9 ml/day, male7 ml/day, female6
1 Ingestion rate cited for European blackbird, Turdus merulab Assumed to be negligible based on feeding habitse Calculated using allometric equation derived by Calder and Braun (1983)
Lethal and sublethal effects of dietary PCBs to mink
Species
Mink
Mink
Mink
Mink
Mink
Mink
Mink
Dose
0.64 mg/kg0.096 mg/kg/day1
2.0 mg/kg0.3 mg/kg/day1
5.0 mg/kg0.75 mg/kg/day1
5.0 mg/kg0.75 mg/kg/day1
20 mg/kg3 mg/kg/day1
31.5 mg/kg4.7 mg/kg/day1
48.5 mg/kg7.3 mg/kg/day1
Exposure
16 weeks; Arochlor1254 mixed in food
4 months; fed dietscontaining Great Lakessalmon
16 weeks; Arochlor1254 mixed in food
8 months; Arochlor1242 mixed in food
8 months; Arochlor1016 mixed in food
35 days; Arochlor 1254fed to rabbits, rabbitsfed to mink
35 days; 1254 mixed infood
Effect
Reproductive failure
Nearly complete reproductive failure
Loss of offspring
Complete reproductive failure
Reduction in kit growth, higher kitmortality
LCjo
LCM
Reference
Platonow and Karstad 1973
Aulerich and Ringer 1977
Ringer 1983
Bleavins et al. 1980
Bleavins et al. 1980
Aulerich et al. 1986
Aulerich e( al. 1986
1 Exposure in mg/kg/day calculated based on an average adult bodyweight of 1 kg and ingestion rate of 0.15 kg/day (Bleavins et al. 1980)
REFERENCES
LIFE HISTORY INFORMATIONBeyer, N., E. Conner and S. Gerould. 1991. Survey of soil ingestion by wildlife. Report on
work funded by U.S. EPA and supervised by Ruth Miller, OPPE. In: WildlifeExposure Factors Handbook. (DRAFT) U.S. EPA.
Bleavins, M.R., R.J. Aulerich and R.K. Ringer. 1980. Polychlorinated Biphenyls (Arochlors1016 and 1242): Effects on survival and reproduction in mink and ferrets. Arch.Environ. Contain. Toxicol. 9:627-635.
Calder, W.A. and EJ. Braun. 1983. Scaling of osmotic regulation in mammals and birds.Am. J. Physiol. 244:R601-R606.
Linscombe, G., N.K. Kinler and R.J. Aulerich. 1982. Chapter 31, Mink (Mustela visori). pp.629-643 In: The Wild Mammals of North America: Biology, Management, Economics.J. Chapman and G. Feldhamer (eds.). Johns Hopkins Press, Baltimore, MD.
RED-WINGED BLACKBIRD LIFE HISTORY INFORMATIONBent, A. C. 1965. Life Histories of North American Blackbirds, Orioles, Tanagers and Allies.
Dover Publication, Inc., New York.
Calder, W.A. and E.J. Braun. 1983. Scaling of osmotic regulation in mammals and birds.Am. J. Physiol. 244:R601-R606.
Case, N.A. and O.H. Hewitt. 1963. Nesting and productivity of the red-winged blackbird inrelation to habitat. The Living Bird, Second Annual of the Cornell Laboratory ofOrnithology, pp. 7-20.
Clench, M.H. and R.C. Leberman. 1978. Weights of 151 species of Pennsylvania birdsanalyzed by month, age and sex. Bull. Carnegie Mus. Nat. Hist. 5. (as cited in Dunning1993}
Dunning, J.B. Jr. 1993. CRC Handbook of Avion Body Masses. CRC Press, Boca Raton, FL.
Kenaga, E.E. 1973. Factors to be considered in the evaluation of the toxicity of pesticides tobirds in their environment. Pages 166-181 in: Environmental Quality and Safety.Global Aspects of Chemistry, Toxicology and Technology as Applied to the Environment.Vol. II. F. Coulston and F. Korte, eds. Academic Press, Inc., New York.
Nero, R.W. 1956. A behavior study of the red-winged blackbird. n. Territoriality. WilsonBull. 68:129-150.
Orians, G.H. 1980. Some Adaptations of Marsh-nesting Birds. PrincetonUniversity Press, Princeton, New Jersey.
Snelling, J.C. 1968. Overlap in feeding habits of red-winged blackbirds and common gracklesnesting in a cattail marsh. Auk. 85:560-585.
PCB TOXICITY INFORMATIONAulerich, R.J., R.K. Ringer and S. Iwamoto. 1973. Reproductive failure and mortality in mink
fed on Great Lakes fish. J. Reprod. Pert., Suppl. 19:365-376.
Aulerich, R.J. and R.K. Ringer. 1977. Current status of PCB toxicity in mink and effect ontheir reproduction. Arch. Environ. Contam. Toxicol. 6:279-292.
Aulerich, R.J., S. Bursian, W. Breslin, B. Olsen and R.K. Ringer. 1985. lexicologicalmanifestation of 2,4,5-, 2,3,6- an 3,4,5-hexachlorobiphenyl and Arochlor 1254 in mink.J. Toxicol. Environ. Health. 15:63-79.
Aulerich, R.J., R.K. Ringer and J. Safromoff. 1986. Assessment of primary vs. secondarytoxicity of Arochlor 1254 to mink. Arch. Environ. Contam. Toxicol. 15:393-399.
Bleavins, M.R., R.J. Aulerich and R.K. Ringer. 1980. Polychlorinated Biphenyls (Arochlors1016 and 1242): Effects on survival and reproduction in mink and ferrets. Arch.Environ. Contam. Toxicol. 9:627-635.
Platonow, N.S. and L.H. Karstad. 1973. Dietary effects of poly chlorinated biphenyls on mink.Can. J. Comp. Med. 37:391-400.
Ringer, R.K., R.J. Aulerich and M. Zabik. 1972. Effect of dietary polychlorinated biphenylson growth and reproduction of mink. American Chemical Society. Division of Water,Air and Waste Chemistry. Preprints of papers presented at 164th National Meeting.12(2): 149-154.
Ringer, R.K. 1983. Toxicology of PCBs in mink and ferrets. In: PCBs: Human andEnvironmental Hazards. P.M. D'ltri and M.A. Kamrin eds. Butterworth Publishers,Woburn, MA.
where I equals the amount of soil in the Gl tract.'S equals the ratio of insoluble ash to dry
contents in the Gl tract, F equals the ratio of insoluble ash to dry contents in fescue (thedominant vegetation in the rat's habitat), and W equals the dry weight of Gl tract contents.
Beyer et al. (1991) used scat samples to estimate the fraction of soil in the diet of anumber of species. The equation for this estimation is slightly more complicated, becausedigestibility and the mineral content of the soil must be included:
Y = (b + cX - bX) / (1 - a + aX) (27)
where Y equals the ratio of ash to dry weight of the scat, b equals the ratio of ash to dryweight in the diet without soil, c equals the mineral content of the soil, and X equals the •tfraction of soil in the diet. To solve for X: . . •
X = (b - Y + aY) / (aY - c + b) (28)
. Beyer et al. tested the accuracy.of this equation in a laboratory study of soil ingestionby feeding mice diets with measured, varying soil components. They found a goodcorrelation between the measured and predicted relationships between the ratio of acid-
insoluble ash to dry weight of scat and the percentages of soil in diet.
3.5.3. Results
Beyer et al. estbnated soil ingestion rates for a variety of animals. Rates for thosespecies which are included as, or related to, the representative species in this Handbook are
listed in Table 3-2. Sandpipers, which feed on mud-dwelling invertebrates, were found tohave the highest rates of soil/sediment ingestion (30,18, and 17 percent of diet respectivelyfor semi-palmated, western and stilt sandpipers). Relatively high soil intakes were estimated
for the raccoon (9.4 percent) and the woodcock (9.1 percent) which also feed on soilorganisms. Soil ingestion was lowest for the white-footed mouse, meadow vole, fox, and box
turtle (<2, 2.4, 2.8, and 4.5 percent, respectively). Box turtles, tortoises and some reptiles,however, have been known to intentionally ingest soil, perhaps for its nutrient content(Kramer, 1973, and Sokal, 1971, both as cited in Beyer et al., 1991).
3-15 " 11/19/91
Table 3-2. Percent Soil in Diet Estimated from Acid-insoluble Ash of Scat (Beyer et al.,
1991).
Species
Semipalmatedsandpiper
Western sandpiper
Stilt sandpiper
Raccoon
Woodcock
Canada goose
Least sandpiper
Box turtle
Fox
Meadow vole
White-footed mouse
Scatsamples1
1
1
1
4
7
23
1
8
7
7
9
% Insoluble AshMean/(Range)
56
42
40
28 (13-50)
22 (6.3-40)
12 *. (3.9-38)
24
18 (3.6-49)
14 (4.8-25)
8.9 (4.2-14)
8.5 (5.7-11)
EstimatedDigest. % ofdiet
70
70
70
70
60
25
70
70
70
55
65
Estimated %soil in diet
30
18
17
9.4
9.1
8.2
7.3
4.5
2.8
2.4
<2
Beyer et al.'s data should be used with caution, however, as error was introduced by
estimating variables in the equation (e.g., digestibility), and by the small sample size theyobtained from some of the smaller animals.
Other studies that considered the soil content in the diet of species related to those
represented in this Handbook are presented in Table 3-3. Arthur and Gates (1988 as cited inBeyer et al., 1991) estimated 6.3 percent soil in the diet of jackrabbrts. The digestive tracts ofshorebirds have been estimated to contain from 10-60 percent sand (Reeder, 1951 as cited inBeyer et al., 1991). Sediment has been found in the stomachs of Ruddy ducks and shovelers(Goodman and Fisher, 1962 as cited in Beyer et al., 1991) and white-footed mice (Garten,
1980). Sediment in the gut of tadpoles inhabiting highway drainages may be responsible for
1 For all the sandpipers, the white-footed mouse, and the meadow vole, scat samplesfrom more than one animal had to be combined to create a sample of sufficient quantity forchemical analysis. This is why only one sample was analyzed for each species of sandpiper.
3-16 11/19/91
high concentrations of lead detected in these organisms (Birdsall et al., 1986 as cited inEeyer et al., 1991).
Table 3-3. Percent Soil in Diet Estimated by Various Sources.
Species
Jackrabbit
Shorebirds
Estimated % soil in diet
6.3
10-60
Citation
Arthur and Gates, 1988
Reeder, 1951
3.5.4. Literature Citedt
*>.
Arthur, W. J., Ill; Alldredge, A. W. (1979) Soil ingestion by muie deer in north centralColorado. "J. Range Manage. 32: 67-70.
Arthur W. J., Ill; Gates, R. J. (1988) Trace element intake via soil ingestion in pronghorns andin black-tailed jackrabbits. J. Range Manage. 41: 162-166.
Eeyer, N.; Conner, E.; Gerould, S. (1991) Survey of soil ingestion by wildlife. Report on workfunded by U.S. ERA and supervised by Ruth Miller, OPPE.
Beyer, W. N.; Stafford, C.; Best, D. (In review) Survey and evaluation of contaminants inearthworms from confined disposal facilities for dredged material in the Great Lakes. 24 pp.
Birdsall, C. W.; Grue, C. E.; Anderson, A. (1986) Lead concentrations in bullfrog Panecatesbeiana and green frog R clamitans tadpoles inhabiting highway drainages. Environ.Pollut. (Ser. A) 40: 233-247. .
Garten, C. T. (1980) Ingestion of soil by hispid cotton rats, white-footed mice, and easternchipmunks. J. of Mammal. 61(1): 136-7.
3-17 11/19/91
Goodman, D. C.; Fisher, H. I. (1962) Functional anatomy of the feeding apparatus in •
waterfowl (Aves: Anatidae), Carbondale, IL: Southern Illinois University Press; 193 pp.
Kramer, D. C. (1973) Geophagy in Terrepene ornate ornate Agassiz. J. Herp. 7: 138-139.
Kreulen, D. A.; Jager, T. (1984) The significance of soil ingestion in the utilization of aridrangelands by large herbivores, with special reference to natural licks on the Kalahari pans.
In: International symposium on herbivore nutrition in the subtropics and tropics (1983:Pretoria, South Africa). Draignall, South Africa: Science Press; pp. 204-221.
Mayland, H. F.; Shewmaker, G. E.; Bull, R. C.. (1977) Soil ingestion by cattle grazing crested*!•
wheatgrass. J. Range Manage. 30: 264-265.
Reeder, W. G. (1951) Stomach analysis of a group of shorebirds. Condor 53: 43-45.
Sokal, 0. M. (1971) L'rthophagy and geophagy in reptiles. J. Herp. 5: 69-71.
3-18 11/19/91
uvvtiBB
1090 King Georges Post RoadSuite 407, Edison, NT 08837 1-201-225-6266
TECHNICAL ASSISTANCE TEAM FOR EMERGENCY RESPONSE REMOVAL AND PREVENTIONETA CONTRACT 68 0-0036
.-TO: Mark Sprenger, EPA/ERT
FROM: Scott Crossman, ERT/TAT
THROUGH: Joe Soroka,
DATE: 18 March 1992
SUBJECT: Literature Search on Mink
TDD: 119201003CPCS: 1070 .DCN: TAT-ll-C-157
Below is the information found for the literature search on mink.
^Reproduction. Mink are sexually mature at the age of 10 months.Mating season varies depending upon latitude and among subspecies,but generally occurs from late February to early April. Littersize ranges from one to eight, averaging about four young perbirth. - '
Habitat. Mink occur in a wide range of wetland habitats including:banks of rivers, streams/ lakes, ditches, swamps, marshes andbackwater areas. Different studies have noted population-specificwetland-habitat preferences among various mink subspecies. Theavailability of den sites is another habitat requirement for mink.Den sites may include old beaver lodges, muskrat bank burrows orhouses. Studies have indicated that den availability may effectmink populations, and as more den sites become available inirJcpopulations may increase.
Population Density. Mink populations vary widely depending uponhabitat quality, den site availability, geographic location andtrapping pressures. The following is a summary of mink populationdensities based upon trapping recdrds and mark and recapturestudies:
Roy F. \Vfeston, Inc.MAJOR PROGRAMS DIVISIONIn Association with Foster Wheeler Enviresponse,and R-E. Sarriera Associates
esource Applications, Inc., C.C, Johnson Malhocra, P.C-,
Mink -opul**ions per 10 ha (24 * res)
Number of Mink
2.5 - 4.0"
1.0'
0.4 - 0.2"
0.4*
0.4"
0.8"
0.3"
Jtobitat Type _
cypress-tupelosouthern-Louisiana swamp"better drained bottomlands"Louisiana brackish marshFresh-water marshMontana river in 1957Montana river in 1958
Author
St. Amant (1959)H
n
M
Palmisano (1971)Mitchell (1961)
n
"Based upon trapping records of the number of pelts per area"Based upon mark and recapture studies
Home Range. Male mink generally have much larger home ranges thanfemale mink. During the winter mink "tend to concentrate theiractivity in a more restricted portion of the home range. Maleactivity throughout their entire home range tends to be maximizedduring the mating season*. Using telemetry data, Gerell (1970)calculated home ranges for adult mink along a stream. He foundadult male mink have home ranges averaging 2,630 meters of streamlength with a range of 1/800 to 5,000 meters. Adult females hadhome ranges averaging 1/850'meters of stream length, but variedfrom 1,000 to 2,800 meters. For both sexes juvenile home rangeswere significantly less. Mitchell (1961) estimated home ranges fortwo adult female mink along the Montana river. The first femalehad a home range of 7.8 ha based upon 7 captures and the secondfemale had a home range of 20.4 ha based upon 10 captures. Henoted that the first female's home range was in optimal habitatcontaining excellent cover, with a small drainage ditch runningthrough it. In comparison the second females home range was in amarginal habitat containing limited cover.
Polychlorinated Biphenyls* Mink have been found to be extremelysensitive to PCBs. Platonow and Karstad (1973) noted in ranchedmink as little as 0.64 ppm of PCS (from contaminated meat) in thediet for 160 days caus«d nearly complete reproductive failure,while 3.57 ppm dietary PCB was lethal to adult mink. Bleavins etal, (1980) found in a 247 day study mink fed concentrations as lowas 20 ppm Aroclor 1242 exhibited 100% mortality.
Cited
„ T * u i e r * a n ' - . 1980. PolychlorinatedBleavins M.R. , R-J- ""Jois and -1242) : "Effects on survival and
biphenyls (Ar°.clorfLfc d ferrets. Arch'. Environm. Contain.reproduction in mi«Toxicol., 9:627-635^ ..„•,- -
,0*a a tivi« patterns of ;Vthe;' mink -Mustela visonGar.ll/ --R'-vW^^uto^ - - -. Schreber in sourner» *•
'"' and populations Vn;a Montana
•*-iana/s fur industry, commercial Wildl.
Palmisano, A.W. 1971. Lox»» ^ wildl- and Fisheries Comm. to U.S.Work Unit Rep of Lou^ Orleans District. Mimeogr.Army Corps of Eng. , **
tad L H 1973. Dietary effects ofS on .in,. Can. J. «. Med. , 37:331-
400. ' - '. -. - - - -. '
329 pp.
couldafter
'L'LTTR.P, CA-
CAN ••*h. A*rtr
J. ROS-
ROUGH.:efibrilU..eetroda.
OSBO-P. S.
; 9* kclae com-
. J
Dietary Effects of Polychlorinated Biphenylson Mink
N. S. Platonow and L. H. Karstad*
I ABSTRACTT** Poisoning occurred In 32 mink fed diets con*(lining meat from cows which had been fed aplychlorinated biphenyl (PCB), Aroelor 1234.Xo live kits were produced and all adult minkfed during a 105 day period of feeding a ra-tion containing 3.57 ppm of PCB. At a level ofi$4 ppm of PCB in ration one of 12 mink pro-duced three kits, all of which died during theQrtt day after birth. Clinical signs were lim-ited to weight loss and passage of black tarry
{frees. The gross lesions seen were yellowishJfiKoIoration of the liver and hemorrhage intoj&e abdominal cavity or gastrointestinal tract.ileroscopic lesions were nephrosis, fatty de-generation and necrosis of the lirer, brainJem a, disseminated intravascular coagulation,tod fibrosis of coronary arteries. It is con-cluded that mink are highly sensitive to small(itntities of PCB fed for an extended periodrf time.
RESUMt
On a empoisonne 32 visons en leur servantme ration qui contenait de la viande de vachesnxquelles on avait donne un diphenyle poly-
ore (PCB), 1'ArocIor 1254. On n'obtint au-in petit et tous les visons adultes moururenticours d'une periode de 105 jours durant la-dle on leur servit une ration contenant 3.57
de PCB. A la concentration de 0.64 ppm
fco«rtr»«nu mt Bi«m«dl«al SeUnea uidolph. Gu«l»h. Out aria.
' B*p«r wm» pm«nt*d tn p»rt at th« Fift«nth Annualdna; *f Ui« Canadian F«d»ralien of Bialoirirai So-
de PCB dans la ration, une femelle, sur un to-tal de 12, mit bas trois petits qni moururent enmo ins de 24 heurw apres lear naissance. Lessignes cliniques se traduisirent par une pertede poids et le passage de feces d'apparencegoadronnee. Les lesions macroscopiques con-aistaient en une decoloration jaunatre du foieet «a des hemorragies intra-abdominales ougastro-intestinales. Les lesions microseopiquesetaient les suivants: nephrose, degenerescencegratsseuse et necrose hepatiqaes, oedeme cere-bral, coagulation intra-rasculaire disserainee etfib rose des arteres coronaires.
L*s auteurs en nennent a la conclusion quele vison est tres vulnerable a I'inpestion pro-longee de petites quantites de PCB.
INTRODUCTION
Polychlorinated biphenyl (PCB) com-pounds are now recog-nized as widespreadenvironmental contaminants of aquatic orterrestrial ecosystems. Since fish and vari-ous byproducts made from domestic animalsare generally included in the diet of ranch-raised mink, it seemed to be important toknow the effects of PCB compounds onmink, when included in the diet for an ex-tended period of time.
MATERIALS AND METHODS
Thirty-two ranch mink approximatelyone year old %vere divided into two groupswith four standard dark males, four pastelfemales, and eif?ht standard dark females ineach group. Each yroup of 16 mink was feda separate ration. Rations were preparedfrom two Jersey cows which had been givenorally ten consecutive daily doses of 1 and10 rngm per kgrn, respeczively of a PCB,
Med- Kl. 37 — October, 1973 391
namely Aroclor 125-4', dissolved in a smallamount of olive oil and ntixed with dairyconcentrate. Twenty-four hours after thelast doses wera given, the csws were killed;fat, musculature, liver and kidneys wereremoved, thoroughly ground and mixed withcommercial mink food cereal at a level of24 c'c cereal. The resulting rations contained0.64 ppm and 3.57 ppm of total PCB, re-spectively.
The cows did not have any clinical, grossor histopathological signs of intoxication.If these animals had been slaughtered at aninspected meat plant, they would withoutdoubt have passed as fit for human con-sumption.
Feeding ad libitum was beyun two monthsbefore the breeding season, i.e. on January7 and continued unti l June 17, 1971 (160days) .
Beginning in early March the 12 femalesin each test diet group were presented dailyfor breeding to one of the four males in thegroup, until the female WM found to be re-ceptive and mating took place. The semen ofeach male, collected by vafinal aspirationfrom a just-bred female, was examined mi-croscopically for motile sperm.
The remaining 101 breeding female minkplus males and mink designated for otherexperiments in the institutional herd fromwhich the test mink were taken were main-tained on a commercial mixed ration, pur-chased locally. They were managed dur ingthe breeding and whelping seasons just aswere the experimental PCB-fed animals.For practical purposes, they were regardedas no-treatment controls. During the courseof the PCB experiments 50 to 60 mink fromthis herd were removed for aleutian diseaseresearch, either killed at once for tissuecultures or maintained on other premisesfor infection experiments.
All of these mink wers examined forgross and histopathological lesions.
Necropsies were performed on experi-mental mink which died and those whichwere killed after PCB feeding was discon-tinued. Blood from the heart and samplesof tissues from the major body organs andpsoas muscle were taken for PCB analysis.Portions of brain, lungs, heart, spleen, li-ver, kidney, intestine, adrenal and reproduc-tive organs were fixed in 10 c formalin,processed for histology by paraff in embed-
'Arocior 1254 (mistur* of chlorinat«4 biphtnrl* contain-ing i-trV chiortn*) WBI ««n*roml)< »apli«4 by MonaanuCanada Ltd.. Toronto. Ontario.
392
TABLE T. Lesions Seen in
FCB > p p m j 3.57No of mink 12Sex FNo of deaths 12/12Mean survival
i days j 74Gross LesionsWeight loss 8/12Hemorrhages 9 '12Yellow Liver 6/12,Microscopic LesionsNephrosis 5/12Liver necrosis 2/12Disseminated intra-
vascular coagu-lation 2/12
Brain edema 3/12Coronary arterio-
sclerosis 0/12
Mink Feed PCB
3.57 0.54
M "~^ - l/ll
96 l.-,4
2 '4 0'123/4 Q ' I 22/4 1'12
1/4 1/122.4 0-12
0 4 l ' I 20 4 0 12
i -: :•All mink in this group survived :o L.\C endfeeding period. 160 days
ding and sectioned at approximatelycrons. Hematoxylin-eosin stain was
0.644
M0 ,4
160
0 40/40 , 4
0,40/4
0/40.4
1 4
of the
6 mi-used
tlher befor-?cd. T'.VO o:-;ed d u r i n g .5 breed wa.fleeting -r.~fas too \vea-:|Ail rr.aie= ;•nars were•£t ra t ion cc:joiy one of ;^educed a I:^is female b.
iv of l i re ..
LI.VICAL Sic:The firs; .T.
[?d the racierr-fch was d=•u:. Deaths::ar. All of :h» .rizon were c-* • f ~ .Lax f ,.G4Ja 12!?rTesnt
routinely. Where necessary for study of !•-sions of blood vessels, selected tissue sec-tions were stained by the allochrome proce-dure ( 5 } .
The PCB were extracted from varioustissues as well as from samples of rationsby the method of Grant et (tl • ? :.-i subse-quently analyzed by the methc- ;. Saschen-brecker and Ecobichon (12) using a Micro-Teck. Model MT-220*, gas chromatograph.equipped with a MNi high temperature elec-tron capture detector. The quantitation ofthe PCB was performed by measuring totalpeak area as the detector response, using anInfotronics, Model 208', automatic digitalintegrator equipped with a baseline track-ing and drift corrector. The PCB wereseparated from DOT and its analogs by themethod described by Armour and Burke( I K
RESULTS
REPRODUCTIONFifteen of the 24 females were bred nor
mally. Of the remaining nine, five on thigher PCE diet which were not bred,
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TAI1I.R II. Mean Concentrations (it stand, dcv.) In ppm of Polychlorlmited lilphenyla (PCH) In Varloim TUauet of Mink Ked Italian*Containing liigretllcnt(ii) Contaminated with PCU
PC1I level NumberIn ration of Period1 Hlood
(ppm) Animals3.57 16 — 1.80 ± 1.42
0.64 2'- • 0.71 ± 0.01
0.64 4 0 0.12 ± 0.02
0.64 4 1 0.10 db 0.05
0.64 4 2 0.24 ± 0.03
0.64 2 3 0.06 ± 0.07
Control(0.30 *0.08) 8 — 0.12 ± 0.08
•Period (in months) after withdrawal of PCD ration"•Died during feeding the ration containing 0.64 ppm of PCU
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4.72 ± 3.31
0.52 ± 0.01
1.36 ± 0.45
0.60 ± 0.26
0.90 ± 0.13
0.33 i 0.01
0.32 ± 0.09
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Tcorpora ieutaa present, ir. various ae-of resrression. Ovarian follicles seamed
;!) be present :n adequate numbers during-je breeding1 period, although ^iany of :he-•oilicJes were undergoing: regressive changes'ji mink which died. Females which died be-fore or at the beginning of the breedir.y pe-:iod had quiescent uteri which were juvenile
1 ji app«aranca. Male n-.ink which died dur-! M March and April had histologicai evi-! jence of sperrr.atoffenesis. These which diedI jf were killed later had testicuiar changes
rrpical of post-breeding- rejrassion.No lesions were observed in the adrenal
jiands.
! TISSUE DISTRIBUTION OF PC3j The concentrations of PCS in various! rissues of principal and control mink are! fiver, in Table II.
In the group of mink fed the higr.er 'eve:PCB, the concentrations :: these corr.-
unds were lowest in the biocd and hiyhesiliver. In the same group, the PCS
concentrations in the brain ware relatively' high, especially when one considers that• ihey were higher than in the skeletal muscle.•;The concentrations of PCB in kidneys were'iijner than in the brain, but lower than inItbe hearts and livers.| In mink fed the ration containing 0.64j?pm of PCB. the PCB concantraticr.s ins tissues were similar in al! organs analyzed.JKcept in blood, where the lowest levels.«re detected. Similarly, in mink fed thejMmmercial control ration, the PCB concen-{trations were nearly identical in the anal-jfied tissues, except in blood.j The resolution by gas-chrorr.atography of'Aroclor 1254 standard as well as extracts oftnrious feed or tissue samples (under the'^erating conditions used in the pr2Si;:it
ydy) resulted in up to 18 distinct peaks,jngure 4 depicts the chromatcjraphic trac-jjg of tissue extracts from rr.ink fed theJ-^tion containing 3.57 ppm PCB as wail as.ie tracing of a standard Aroclor 1254.••Vote the decrease of early emerging peaks
'•f tissue extracts, disappearance of peak Sud increase of late emerging peaks.
The data depicted in Fig. 3 and Fig. 6show the ratios between the ir.aan percent-''?ss of peaks from each tissue extract ofroups giver. 3.57 ppm and 0.-34 ppm1 PCB"fl ths percentages of the corresponding"•sak of the standard Aroclor 1234. The base
j-'ne in each cssa is '.:n-i;y. The zaro valiseindicates the disappearance of the peak: thi
Vol. 37 —October, 1973
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values biv.veen uni ty ar.c zaro ir.iicate the. percsr.tejs decrease of :he peak: the valueshigher thin unity incicata the ir.ireases inpercer.:aj? above the value of the jtandarti.
The percentage reductions of the firs:eight pei!-:s and tha percentage increases inthe 3ubs5:uer.: peaks in these "vo figuresare avic*r.t. The first eight emerrir.? peaksware 3:?r.!::car.::y raz-cad in s.'.'. tijsuas.except ir. :!ocd, whera :he reductiir. was notalways =!»-::icar.:. The -i*cand sijht peakswere always sisr.iricar.ti;.- incraased pro-por;:or.a!!y *o the Arcc'.c:- 125-4 standard.These chir.jea were most pror.ov.-:sd in th*liver ar.i '.333: so :r. the blood. Peak 3 wasred'.:esc ~~ zero ir. all tissues, except ir.b'.coc: r:*ik 7 '.va* tha r.axt r.cs: s:rony:yradussd. I- peaks err.arjir.j aft*:- pea-it S.:hs prt^rrticr.ai ~ir:3r.ta?» ir.craase wasr.aarly i:rs:t:y ranted to tha retention tirr.2.i.i. peak I-j increased r.rre than peak £.
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This -3r.der.cy was not observed in blood. Onail chromatograms a substantial percent-a?* '-.crease was noted in peaks 1. and 13.These two peaks are below 0.1* m theArcclor 1254 mixture.
F'gure " shows the ratios expressed si-m-a-iv M Fig. 3 and Fiff. 6 but give the ra-tios :cr both PCB rations fed. anc for com-parisor. :he values of livers of the group fed
RSPRCDVCTIOX AND HEALTH OF COXTSOLMINX
The 101 mink which received the commer-cial ration and which were regarded asno-treatment controls, produced only anaverage of 1,31 kits per female bred (satis-factory production is considered to be fourkits per female1).
When the commercial ration was analyzedthe following results were obtained. Thedata represent means of six samples takenat intervals during the experiment.
No outbreak of infectious disease wasdiagnosed in the hard during these experi-ments. Lesions sirr.i'.ar to those of the prir.-
393
cipal PCB fed groups were net found in theseveral dozer, control mink killed for tissuculture purposes and used ir. aieutian cis-ease research.
DISCUSSION
From the impaired reproduction ar.dmortality observed, and the tissue concen-trations of PCB found, it can be concludedthat the mink were very susceptible to theeffects of PCB compounds as dietary con-taminants. The clinical and gross and mi-croscopic tissue abnormalities were rathernonspecific, so that diagnosis ir. ::::• ofnaturally occurring PCB poisoning wouldbe very difficult. Reproductive failure.weight loss and tarry droppings couldsuggest either starvation or aieutian dis-ease, a comrr.on disease of mink. The PCB-fed mink may have been on an inadequatenutritional plane, but if so. it was not be-cause the diet lacked essential nutrientsbut rather that the mink lost appetite anddid not eat enough of the feed that wasput before them. Aleutian disease wouldbe ruled out on histopathoiogy, however,since the severe hepatitis and n-;V::t:s ofaieutian disease, • marked by pic^Seratlonof plasma ceils, was not seen in the PC3-poisoned mir.k.
The occurrence of liver damage in minkwhich also developed hemorrhagic tenden-cies suggests a breakdown in hemostasisin which prathrombin deficiency may be acausative factor. Blood coagulation studiesshould be rr.ade in future research on PCBpoisoning in mink. Splenic enlargementand increased numbers of megakaryocytMin the splesn may represent compensatoryresponses ir.. attempts to tr.a:r^~:n herr.o-stasis. The cause of the cis:;....--';a«d, !n*travascular coagulation seer, in the tare*mink is ur.known. Hormonal effects ^ofPCB may have been responsible for tr.ereproductive failure. The presence of cor-pora lutea ir. the ovaries of the mink waicfldied during or immediately after the nor-mal gestation period indicates that ovuia-tion did occur but that gestation did no-continue to term. , .. „
Fatty degeneration of the liver ar.o '«•necrosis has beer, described in otnercies treated with PCB ccrr.?r;r.cs.generative kidney lesions a.s: •"3
-link^•.den-stasis;.- be a-.urfies; PCB-ement-cytessatoryherno-
thre*cts of... the-; cor-which
:e nor-ovula-
:id not
, been
M«J-
in other species by some rk-Platonow et a.1 (9) have deacribt^. pe-
'ivascular hemorrhages around coronaryin the hearts of chickens fed Aro-
The coronary arterial lesions seen inthree of the mink in these experiments are^explained but thought to be related to thepCB feeding, since lesions of this type have*ot been seen by the authors previously.Their appearance in two of three minkJhich died between days 105 and 129 ofpCB feeding and in one of four mink killedon day 160 may mean that lesions in theseJital arteries develop as a late response totow-level PCB ingestion. This should be in-vestigated further in mink and also inother species.JThe poor reproduction in the no-treat-*ent controls may be referable to the low£o.3 ppm) but significant amounts of PCBfound in the control commercial ration. Thecontrols, therefore, actually were an un-planned lower PCB treatment group ofmink. Lesions referable to the PCB con-fent of the control ration were not seen inthe mink from this herd which were usedfor research on aleutian disease duringtte course of these PCB experiments. The£w concentrations of dieldrin as well asDDT and its analogs and metabolites in thecontrol ration are for all practical purposesif no toxicological significance.Tit has been suggested that PCB affectenzyme systems that metabolize severalKX hormones, which ultimately leads to
fnproductive failure. Thus, Risebrough etlit (11) reported that the PCB induce theIfctivity of hydroxylating enzyme whichMetabolizes estradiol. The estrogenic acti-Jnty of various PCB mixtures upon the rat
cerus was described by Bitman and Cecil2). Platonow and Funnel (7, 8) reportedut chronic feeding of PCB in cockerelssuited in anti-androgenic effects, asuessed by decreased development of tes-cles and comb. These signs appeared asirly as one week after the start of con-nuous feeding of a PCB-contaminatedlet.[it was recently demonstrated by No-icki and Norman (6) that the post-mito-umdrial hepatic fraction (microsomes -f-Jtosol) from PCB-treattd cockerels and*Hets metabolized testosterone, estradion-
and 4-androstene-3, 17-dione at in-Fused rates. The metabolic rates of theseiree natural steroid hormones were in-
ed approximately three-fold over•eated birds. Platonow, Liptrap and
kl. 37 — October, 1973
Geissinger (8) reported on the t t oforal administration of PCB (Aroclor1254) in the adult boar. The doses of PCBwere such that no gross or histological le-sions could be seen. However, the presenceof significant biochemical alterations wererecorded. These changes consisted in dras-tic reduction of urinary levels of two gona-dal hormones: dehydroepiandrosterone andestrogsn, indicating that PCB have a de-leterious effect upon reproductive activity.The primary site of o,p'-DDD inhibition ofACTE-induced steroidogenesis in the adre-nal cortex appears to be on the ACTH-regulated intramitochondrial conversion ofcholesterol to pregnenolone (4). Since thechemical formulae of PCB's are similar tothat of DDT, it is possible that the hor-monal disturbances due to PCB are locatedat the same site. At least this is a prob-able site of PCB effect in the boar, be-cause pregnenoione is a precursor of de-hydroepiandrosterone in steroidogenesis.
In comparison to domestic animals (8,9, 10, 13) mink seem to extensively meta-bolize the PCB, Aroclor 1254 used in thisexperiment. The changes occurring on thechroma to grams of various tissue extractsindicated significant reduction of the firsteight peaks, with simultaneous increasesof the peaks emerging thereafter. Theorder of appearance of peaks on the FCBchromatogram is related to the extent ofchlorination of biphenyls. Thus, mono-chlorbiphenyls appear before dichlorobi-phenyls, trichlorobiphenyls before tetra-chlorobiophenyls, etc.. and decachloro-biphenyl (the completely chlorinatedbiphenyl) emerges as the last peak. Thereduction of peaks of lower retention in-dicates that mink can metabolize PCB oflower chlorination numbers. Since this re-duction in faster emerging peaks Is muchless pronounced in the cow (13), pig (10),or chicken (8), one might suggest that theextent of PCB metabolism is related to theextent of susceptibility to the toxic *»f *ectof PCB. Mink appear to be more susceptibleto PCB than the domestic animals studied.However, further studies are required toconfirm or reject the above hypothesis.
ACKNOWLEDGMENTS
The authors are grateful to Drs. JoanBudd and Peter Lusis for assistance in ne-cropsies, to Mrs. N. Y. Chen for performing
399
the analyses of PCB, and to Mr. Hugh Bel-cher for care of the experimental mink. Thehistopathology was done while the juniorauthor was a visiting professor at UtahState University.
REFERENCES
1 ARMOUR. J- A. mild J. A. BLTIKE. M«thod for ««.ptratinc polrehlorinattd biph*nyt« from DOT andtla anaion- J. Aaa. off. anairt. Ch«m. S3: T81-.S8.19TO2. BITMAN. J. an* H. C. CECIL. Eitroc«nic activitya. DOT analoru* and polyealoriaatMl biphanyla. J.asrie. Fd Ch«m. II: "W-Uis,,1*7* . . „
3. GRANT. D. L, W. E. J. FHIU.IFS »nd D. C.VlLLENEL'VE, M«aboliam of a poly eh Ion nai«d bi-ph«nyl lAroelor 12541 mixtur* ia the r*L. BulU «n-vir. Contam. Toxic. I: n.2-112. 1971.A. HART. M, M. «n«( J. A. STRAW, studio on th« «»of keuon at o.p'-DDD in to* do* »dr«n»l corttx. t.Inhibition of ACTH-m«dl««d prwnenolont iynth«i».St.nsidi IT: SS9-S7J. 19TV.
5. LILIE, R. D. Alloehrem* lUin. Am. J. elm. Fmth.11: 494. 1951.
S. NOWICKI. H. G. and A. W. NORMAN.h* pa tie m«tabaliam of tottau17-dion*. and «atndioM70 tawith DOT or PCB. Steroida 19: W-1M. ijV
7. PLATONpW, N. S. aad H. S. FUNNCLL, ABIL.dro«cnic-lika effect of polyehkirinatad blo«en»ucockerel*. Vet. Rec. II: 109-110. 1911.
8. PLATONOW. -N. S. and H. S, FUNNELL. TH di*.iributioa aad aom« affect* of polyehlonnaied hi».-nyla lAroelor 12S4) in eoek«r«ii dur;--fMdioc trial. Can. J. eomp. M«d. 1C
9. PLATONOW. N. S-. L. H. KARST. 'SASCRENBRECKXR. Ttaau« dlatriL...;achlorinated binhcnyia (Aroclor 1154, ,nRalation to th« duration of •xpoaur* and __tion on patholocy. Caa. J. com p. M*d. 37: (O-it
10. PLATONOW. N. S, R. M. LlPTRAp ..U IT ft"GEISSINGER. Tht dUtributton and *xemloa .(polyehlorinatad biphenyla lAroelor 12S4) andcf/«e: on urinary gonadal lUrnid l«v«l< in tbtBulL tnvir. Coaum. Toxic. I: 358-3(3. 1972.
11. R1SEBROUGB. R. Wv P. REICHE. D. B. PEAKALL. S. G. HERMAN and H. N. KIRVZN. PoiV.chlorinattd bioh«tiyta in U>* flobai acoayttam. W..tur*. Load. 220: lOfS-U02. IMS.
12. 3ASCHENBRECXC&. P. W. aad D. J. ECOBICRONEjttraction aad ru ehromatoftrapbie aaalnu ifchlorinated inaacticfdw front animal tivtuc. J. acnrFd Ch«m. IS: US-170. 1H7.
13. SASCHEXBRECKER. P. W^ N. S. PLATONOW tMH. S. FUNNELL. Metabolic itudy of ix>iyeaiortnat*4bioncnyit in laetating eowi. Pro*. O.rv Fid.Soc. II: <«. 1971.
