Richard J. Aulerich

Current status of PCB toxicity to mink, and effect on their reproduction

Experiments were conducted from 1968 to 1974 to investigate reproductive complications and mortality in mink fed Great Lakes coho salmon and to ascertain the effects of polychlorinated biphenyls (PCBs) on this fur bearer. The results of mink feeding trials indicated that coho salmon, as such, were not responsible for the loss of reproduction in the adult, or the kit mortality. Mink diets that contained other species of Great Lakes fish caused similar reproductive complications, but to a lesser degree.Rancidity, mercury poisoning and chlorinated hydrocarbon pesticide contamination of the fish were all discounted as being responsible for the problem. The clinical signs and lesions noted in mink that died while receiving diets that contained Lake Michigan coho salmon were very similar to those observed in mink fed on rations that contained supplemental PCBs. These included anorexia, bloody stools, fatty liver, kidney degeneration, and hemorrhagic gastric ulcers. Analyses of tissues from mink that died when fed 30% Lake Michigan coho salmon or 30 ppm supplemental PCB diets showed similar PCB residues.PCB toxicity experiments revealed that mink are very sensitive to these compounds and that the lethal dose varied inversely with the chlorine content of the PCBs although only Aroclor 1254 exerted a detrimental effect on reproduction when fed at a low level (2 ppm) for 8 months. The reproductive failure encountered in feeding mink Lake Michigan coho salmon and Aroclor 1254 was shown to be of a non-permanent nature.

Polychlorinated biphenyls (Aroclors 1016 and 1242): Effects on survival and reproduction in mink and ferrets

Diets that contained various levels of supplemental Aroclor® 1242 or Aroclor® 1016 were fed to mink and ferrets to investigate the chronic toxicity of these PCBs in two closely related species.In mink, Aroclor 1242 was found to be more toxic than comparable or higher levels of Aroclor 1016. The Aroclor 1242 diets caused complete reproductive failure at levels as low as five ppm of the diet. Aroclor 1016 impaired reproduction less than Aroclor 1242. Although fewer females whelped and the four-week kit weights were less than the control animals, no outward signs of abnormalities beyond their smaller size were found in the kits whelped and nursed by dams fed Aroclor 1016.Ferrets were more resistant to the effects of either PCB mixture than were the mink, as noted by the lower mortality rate on the Aroclor 1242 diet and the almost normal level of reproduction on the Aroclor 1016 diet. Feeding Aroclor 1242 at 20 ppm resulted in complete reproductive failure, but was not fatal to adult ferrets. This finding is in sharp contrast to the 100% mortality of adult mink fed the same level. Although the chlorine content is similar in both compounds, Aroclor 1242 has a higher percentage of molecules with five or more chlorines per biphenyl. This difference in higher substituted biphenyl isomer content and/or the reduced levels of contaminants in the Aroclor 1016 mixture may be of major importance in evaluating the toxicity of these compounds.

Dietary exposure of mink to carp from Saginaw Bay, Michigan. 1. Effects on reproduction and survival, and the potential risks to wild mink populations

Carp (Cyprinus carpio) collected from Saginaw Bay, Michigan, containing 8.4 mg total polychlorinated biphenyls (PCBs)/kg and 194 ng of 2,3,7,8-tetrachloro-dibenzo-p-dioxin equivalents (TEQs)/kg, were substituted for marine fish at levels of 0, 10, 20, or 40% in the diets of adult ranch mink (Mustela vison). The diets, containing 0.015, 0.72, 1.53, and 2.56 mg PCBs/kg diet, or 1.03, 19.41, 40.02, and 80.76 ng TEQs/kg diet, respectively, were fed to mink prior to and throughout the reproductive period to evaluate the effects of a naturally-contaminated prey species on their survival and reproductive performance. The total quantities of PCBs ingested by the mink fed 0, 10, 20, or 40% carp over the 85-day treatment period were 0.34, 13.2, 25.3, and 32.3 mg PCBs/mink, respectively. The corresponding quantities of TEQs ingested by the mink over the same treatment period were 23, 356, 661, and 1,019 ng TEQs/mink, respectively. Consumption of feed by mink was inversely proportional to the PCB and TEQ content of the diet. The diets containing Saginaw Bay carp caused impaired reproduction and/or reduced survival of the kits. Compared to controls, body weights of kits at birth were significantly reduced in the 20 and 40% carp groups, and kit body weights and survival in the 10 and 20% carp groups were significantly reduced at three and six weeks of age. The females fed 40% carp whelped the fewest number of kits, all of which were stillborn or died within 24 hours. Lowest observable adverse effect levels (LOAEL) of 0.134 mg PCBs/kg body weight/day or 3.6 ng TEQs/kg body weight/day for adult female mink were determined. The potential effects of exposure of wild mink to contaminated Great Lakes fish were assessed by calculating “maximum allowable daily intakes” and “hazard indices” based on total concentrations of PCB residues in several species of Great Lakes fish and mink toxicity data derived from the study.

Feeding great lakes fish to mink: Effects on mink and accumulation and elimination of PCBS by mink

The effects of feeding Great Lakes fish or fish products (carp, sucker, perch scraps, whitefish racks, and alewife fishmeal) to mink were studied. Growth and furring of mink were normal for all species of fish tested. However, mink fed carp failed to reproduce, and the reproductive performance and/or kit survival in groups fed the perch, whitefish, and sucker were inferior to the control. Only the alewife fishmeal diet supported reproduction and kit survival comparable to the control. PCB residues (as Aroclor 1254) accumulated in mink subcutaneous body fat to as much as 38 times the dietary level, while some individual congeners accumulated up to 200 times. The half-life of PCB in mink adipose tissue was determined to be 98 d. The toxicity of PCBs derived from Great Lakes fish was greater than that observed in previous studies with mink fed comparable levels of technical-grade PCBs.

Contaminants in fishes from Great Lakes-influenced sections and above dams of three Michigan rivers. II: Implications for health of mink

Populations of mink (Mustela vison) have declined in many areas of the world. Such declines have been linked to exposures to synthetic, halogenated hydrocarbons. In the Great Lakes region, mink are fewer in areas along the shore of the Great Lakes and their tributaries where mink have access to fish from the Great Lakes. Recently, there has been discussion of the relative merits of passage of fishes around hydroelectric dams on rivers in Michigan. A hazard assessment was conducted to determine the potential for adverse effects on mink, which could consume such fishes from above or below dams on the rivers. Concentrations of organochlorine insecticides, polychlorinated biphenyls (PCBs), 2,3,7,8-tetrachloridibenzo-p-dioxin equivalents (TCDD-EQ), and total mercury were measured in composite samples of fishes from above or below hydroelectric dams on the Manistee and Muskegon Rivers, which flow into Lake Michigan, and the Au Sable River, which flows into Lake Huron. Concentrations of organochlorine insecticides, PCBs, and TCDD-EQ were all greater in fishes from below the dams than those from above. Concentrations of neither organochlorine insecticides nor mercury in fishes are currently a risk to mink above or below the dams. All of the species of fishes collected from downstream of the dams contained concentrations of PCBs and TCDD-EQ, which represent a hazard to mink. The hazard index for PCBs was less than one for the average of all species from the upstream reaches of the Manistee and Au Sable Rivers, but not the Muskegon. The hazard index (concentration in fish/NOAEC) was greater than 1 for all of the species collected from below the dams, in all three rivers. The greatest hazard index was observed for carp (Cyprinus carpio) downstream on the Muskegon River. Because the concentrations of PCBs used in the hazard assessment were corrected for relative toxic potencies, the hazard ratios based on PCBs should be similar to those based on TCDD-EQ. This was found to be true. Thus, either total PCBs or TCDD-EQ could be used as the critical toxicant in the hazard assessment. However, if uncorrected concentrations of PCBs, expressed as Aroclors®, were used in the hazard assessment, the toxicity of the weathered mixture would have been underestimated by approximately five-fold, and, in that instance, TCDD-EQ would be the critical contaminant for the hazard assessment. The average maximum allowable percentage of fish from above the dams, which would result in no observable adverse effects of TCDD-EQ, was 70%. Based on the average TCDD-EQ concentrations in the fishes, an average of 8.6% of the diet could be made up of fishes from below dams on the rivers. The most restrictive daily allowable intakes were for carp on the Muskegon and steelhead trout (Onchorhyncus mykiss) on the Manistee Rivers. Only 2.7% of the diet could be made up of these two species from influenced portion of the Au Sable River, they would be exposed to 390 μg PCBs and 8.55 ng of TCDD-EQ per day, respectively (Giesy et al. 1994b). Thus, it would take 15.1 or 77 days for mink to receive their total annual dose of PCBs or TCDD-EQ, respectively. At least for chinook salmon, the critical contaminant for the purposes of hazard assessment would be total concentrations of PCBs. Consuming chinook salmon for as little as 2 weeks would deliver the annual allowable dose of PCBs to mink.

Effects of 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) injected into the yolks of chicken (Gallus domesticus) eggs prior to incubation

The yolks of White Leghorn chicken (Gallus domesticus) eggs were injected prior to incubation with either 3,3′,4,4′,5- pentachlorobiphenyl (PCB 126) at doses ranging from 0.1 to 12.8 μg/kg egg or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at doses ranging from 0.04 to 0.64 μg/kg egg. Chicks were subjected to necropsy within 24 h of hatching. The brain, bursa, heart, liver, and spleen were removed and weighed. Assessment of the rate of hatching indicated an LD50±S.E. of 2.3±0.19 μg/kg egg (7.1±0.58 nmol/kg egg) for PCB 126 and 0.15±0.012 μg/kg egg (0.47±0.037 nmol/kg egg) for TCDD. No significant differences in the incidence of developmental abnormalities (structural defects and edema) were observed in TCDD-exposed embryos, while PCB 126 caused significantly more developmental abnormalities at 3.2, 6.4, and 12.8 μg/kg egg than the vehicle control. PCB 126 caused lower hatchling weights and greater relative brain, heart, and liver weights when compared to the vehicle control group at a dose of 3.2 μg/kg egg which is greater than the LD50. TCDD at 0.08 μg/kg egg caused relative bursa weights to be less than those of the vehicle control. A toxic equivalency factor (TEF) of 0.07 was determined for PCB 126 in relation to TCDD based on overt lethality.

Toxicity of 3,4,5,3',4',5'-hexachlorobiphenyl to mink.

Diets supplemented with 0.01 or 0.05 ppm (mg/kg) of 3,4,5,3′,4′,5′-hexachlorobiphenyl (345-HCB) were fed to mink to investigate the toxicological manifestations of this toxic polychlorinated biphenyl congener in a sensitive species. Dietary exposure of mink to 0.05 ppm 3,4,5,3′,4′,5′-hexachlorobiphenyl for 135 days resulted in 50% mortality while no deaths occurred on 0.01 ppm 345-HCB. Clinical signs of toxicity included anorexia, bloody stools, disrupted molting patterns, and thickened, elongated and deformed nails. Ascites and gastric ulcers were present in animals that died. Statistically significant increases in liver, kidney, and adrenal gland weights were found in the 345-HCB-treated mink. Decreases in total and free triiodothyronine concentrations were observed in mink fed the 345-HCB-treated diets and total thyroxine was decreased in the mink fed 0.05 ppm 345-HCB. No consistent histopathologic lesions were found in the thyroid or adrenal glands of the 345-HCB-treated mink, nor were there any statistically significant differences between the 345-HCB-treated and the control mink in serum epidermal growth factor levels, plasma 17β-estradiol and progesterone concentrations, hepatic aminopyrine N-demethylase, and benzo(α)pyrene hydroxylase activities, hypothalamic norepinephrine, dopamine, and serotonin concentrations or in the incorporation of (3H) thymidine by concanavalin-A-stimulated lymphocytes.

Contaminants in fishes from great lakes-influenced sections and above dams of three Michigan Rivers. I: Concentrations of organo chlorine insecticides, polychlorinated biphenyls, dioxin equivalents, and mercury

Fishes of the Great Lakes contain hazardous chemicals such as synthetic halogenated hydrocarbons and metals. These fish can move from the lakes into the Great Lakes tributaries of Michigan. In doing so, they transport concentrationsof contaminants which may represent a risk to wildlife. Concentrations of mercury (Hg), total polychlorinated biphenyls (PCBs), 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQ), total DDT complex, aldrin, endrin, dieldrin, heptachlor, heptachlor epoxide, lindane, hexachlorobenzene, cis-chlordane, oxychlordane, endosulfan-I, methoxychlor, trans-chlordane, and trans-nonachlor were determined in composite samples of fishes from above and below Michigan hydroelectric dams, which separate the fishes which have access to the Great Lakes from fishes that do not. Mean concentrations of total PCBs, TCDD-EQ, DDT, and most of the other pesticides were greater in composite samples of six species of fishes from below than above the dams on the Au Sable, Manistee, and Muskegon Rivers. Concentrations of mercury, were the same or greater above the dams than below. However, this difference was statistically significant only on the Au Sable. Mercury concentrations ranged from less than 0.05 mg/kg to 0.73 mg Hg/kg, ww. Total concentrations of PCBs ranged from 0.02 to 1.7 mg/kg, ww. Concentrations of 2,3,7,8-tetrachlordibenzo-p-dioxin equivalents varied among fishes and locations. The concentrations of TCDD-EQ ranged from 2.4 to 71 μg/kg, ww, with concentrations in carp being the greatest. Concentrations of TCDD-EQ were greater than the concentrations which would be expected to occur, due solely to the presence of polychlorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF), and technical mixtures of PCBs.

Toxic effects of dietary polybrominated biphenyls on mink.

Serial levels of fireMaster® FF-1, a commercial mixture of polybrominated biphenyls (PBB), and tissues from chickens and a cow that had previously consumed PBB were fed to mink to ascertain the chronic effects of the commercial and “metabolized” form of this compound on mink. Diets that contained 6.25 ppm (or more) PBB were lethal to adult mink within 10 months. One to 2.5 ppm dietary PBB fed for 9 months had an adverse effect on litter size, kit weight at birth, and kit survival. The data suggest that the PBB derived from contaminated beef and poultry was more toxic than the original PBB. The clinical signs of PBB poisoning in mink were food rejection, weight loss, an unthrifty appearance, and fatty infiltration of the liver. Based on these findings, mink must be considered highly susceptible to PBB toxicity. PBB residue levels 60 times the amount in the diet were found in the adipose tissue of the PBB-treated animals.

Dietary exposure of mink (Mustela vison) to fish from the Housatonic River, Berkshire County, Massachusetts, USA: Effects on reproduction, kit growth, and survival

We evaluated the effects of feeding farm-raised mink (Mustela vison) diets containing polychlorinated biphenyl (PCB)-contaminated fish from the Housatonic River (HR; Berkshire County, MA, USA) on adult reproductive performance and kit growth and survival. Diets contained 0.22-3.54% HR fish, providing 0.34-3.7 microg total PCBs (TPCB)/g feed wet wt (3.5-68.5 pg toxic equivalence [TEQ]/g). Female mink were fed diets before breeding through weaning of kits. Twelve kits from each treatment were maintained on their respective diets for an additional 180 d. Dietary PCBs had no effect on the number of offspring produced, gestation period, or other measures of adult reproductive performance. Mink kits exposed to 3.7 microg TPCB/g feed (68.5 pg TEQ/ g) in utero and during lactation had reduced survivability between three and six weeks of age. The lethal concentrations to 10 and 20% of the population (LC10 and LC20, respectively) were estimated to be 0.231 and 0.984 microg TPCB/g feed, respectively. Because inclusion of PCB-contaminated fish that composed approximately 1% of the diet would reduce mink kit survival by 20% or more, it is likely that consumption of up to 30-fold that quantity of HR fish, as could be expected for wild mink, would have an adverse effect on wild mink populations.