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This study evaluated the effects of dietary methylmercury (MeHg) on immune system development in captive-reared nestling American kestrels (Falco sparverius) to determine whether T cell-mediated and antibody-mediated adaptive immunity are targets for MeHg toxicity at environmentally relevant concentrations. Nestlings received various diets, including 0 (control), 0.6, and 3.9 µg/g (dry wt) MeHg for up to 18 d posthatch. Immunotoxicity endpoints included cell-mediated immunity (CMI) using the phytohemagglutinin (PHA) skin-swelling assay and antibody-mediated immune response via the sheep red blood cell (SRBC) hemagglutination assay. T cell- and B cell-dependent histological parameters in the spleen, thymus, and bursa of Fabricius were correlated with the functional assays. For nestlings in the 0.6 and 3.9 µg/g MeHg groups, CMI was suppressed by 73 and 62%, respectively, at 11 d of age. Results of this functional assay were correlated with T cell-dependent components of the spleen and thymus. Dose-dependent lymphoid depletion in spleen tissue directly affected the proliferation of T-lymphocyte populations, insofar as lower stimulation indexes from the PHA assay occurred in nestlings with lower proportions of splenic white pulp and higher THg concentrations. Nestlings in the 3.9 µg/g group also exhibited lymphoid depletion and a lack of macrophage activity in the thymus. Methylmercury did not have a noticeable effect on antibody-mediated immune function or B cell-dependent histological correlates. We conclude that T cell-mediated immunosuppression is the primary target of MeHg toward adaptive immunity in developing kestrels. This study provides evidence that environmentally relevant concentrations of MeHg may compromise immunocompetence in a developing terrestrial predator and raises concern regarding the long-term health effects of kestrels that were exposed to dietary MeHg during early avian development.

American kestrels (Falco sparverius) were fed meat diets containing 0, 3, 6, or 12 ppm (dry weight) methylmercury chloride. Birds fed the 12-ppm diet started to show signs of neurotoxicity after 26 days and all died in 39-49 days. One male kestrel fed the 6-ppm diet died after 75 days of exposure and several others showed signs of neurotoxicity after 45 days. None of the birds fed the 3-ppm diet died or showed signs of toxicity. After 59 days of exposure, mercury concentrations in the liver, kidney, and blood of nonreproducing kestrels increased with increasing dietary concentration. Tissue concentrations of mercury also steadily increased over time in birds fed diets with 6 ppm mercury, which were necropsied at 8, 15, 29, or 59 days of exposure, reaching mean total mercury concentrations of 57, 46, and 45 ppm (wet weight) at 59 days in the liver, kidney, and whole blood, respectively. Two pairs of kestrels at each dietary concentration were allowed to breed. Eggs averaged 8.3 and 18.1 ppm (wet weight) total mercury from birds fed 3- and 6-ppm diets, respectively. Feathers grown during mercury exposure contained high concentrations of mercury: Birds fed 3- and 6-ppm diets contained 275 and 542 ppm total mercury, respectively.

Certain contaminants alter normal physiological function, morphology, and behavior of exposed organisms through an endocrine mechanism. We evaluated how the herbicide linuron, an endocrine-active compound, affects physiological parameters and secondary sex characteristics of the American Goldfinch (Carduelis tristis). When administered at relatively low doses (control, 1.0, 4.0, and 16.0 μg linuron per gram of body mass per day), linuron delayed prealternate molt progression in a dose-dependent manner. At the high dose level, linuron exposure lowered hematocrit and female plasma thyroxine concentrations and increased body mass. Neither plasma testosterone concentrations nor the color of plumage or integument of birds in the treatment groups were different from those of the control group. Overall, the physiological effects that were measured suggested disruption of thyroid function. These results highlight the importance of continual monitoring of avian populations for potential effects of exposur...

The Tri-State Mining District (Oklahoma, Kansas, and Missouri) is contaminated with Pb, Cd, and Zn from mining, milling and smelting. Metals have been dispersed heterogeneously throughout the District in the form of milled mine waste ("chat"), as flotation tailings and from smelters as aerial deposition or slag. This study was conducted to determine if the habitat has been contaminated to the extent that the assessment populations of wild birds are exposed to toxic concentrations of metals. American robins (Turdus migratorius), northern cardinals (Cardinalis cardinalis), and waterfowl had increased Pb tissue concentrations (p0.05) compared with Pb tissue concentrations from reference birds, and the exposure of songbirds to Pb was comparable with that of birds observed at other sites severely contaminated with Pb. Mean activities of the Pb-sensitive enzyme delta-aminolevulinic acid dehydratase (ALAD) were decreased by50% in red blood cells in these birds (p0.05). Several birds had tissue concentrations of Pb that have been associated with impaired biological functions and external signs of poisoning. Cadmium was increased in kidneys of songbirds (p0.05), but no proximal tubule cell necrosis associated with Cd poisoning was observed. Zinc concentrations in liver and kidney of waterfowl were significantly higher (p0.05) than reference values. The increased environmental concentrations of Zn associated with mining in the District accounted for the pancreatitis previously observed in five waterfowl from the District. The District is the first site at which free-flying wild birds have been found to be suffering severe effects of Zn poisoning.

In order to reveal patterns of reproductive aging in birds we focus on a short lived species, the Japanese quail and the American kestrel, which has a life span of medium length. Quail have been studied extensively in the laboratory as models for understanding avian endocrinology and behavior, and as a subject for toxicological research and testing. In the lab, Japanese quail show age-related deterioration in endocrine, behavioral, and sensory system responses; the American kestrel is relatively long lived and shows moderate evidence of senescence in the oldest birds. Using data collected from captive kestrels at the Patuxent Wildlife Research Center, a database was designed to document selected parameters over the life cycle of the kestrels. Life table data collated from many species indicate that longer lived species of birds show senescence in survival ability but this pattern has not been established for reproductive function. We suggest that useful comparisons among species can be made by identifying stages in reproductive life history, organized on a relative time scale. Preliminary data from quail and kestrels, admittedly only two species, do not yet indicate a pattern of greater reproductive senescence in longer-lived birds.

Energy budgets have proven to be a valuable tool for predicting life history from physiological data in terrestrial vertebrates, yet these concepts have not been applied to the physiological effects of contaminants. Contaminants might affect energy budgets by imposing an additional metabolic cost or by reducing the overall amount of energy taken in; either process will reduce the energy available for production (i.e., growth or reproduction). This study examined whole animal energetic effects of polychlorinated biphenyl (PCB) exposure in white-footed mice (Peromyscus leucopus). Exposure to PCBs is known to reduce concentrations of plasma thyroid hormones, and thyroid hormones exert strong control over the rate of energy metabolism in mammals. Peromyscus leucopus that were proven breeders were fed PCBs in their food at 0, 10, and 25 ppm. Through lactation, offspring were exposed to PCB from conception and were maintained on the maternal diet to adulthood. No effects were seen on energy metabolism (O2 consumption, measured in adulthood) or on growth, but there were large dose-dependent decreases in thyroid hormone concentrations, particularly T4. The apparent disparity in our data between unchanged metabolic rates and 50% reductions in T4 concentrations can be rationalized by noting that free T3 (the fraction not bound to plasma protein) in treated mice was not significantly different from controls and that metabolism is most strongly influenced by free T3. Overall, this study did not demonstrate any energetic consequences of PCB exposure in P. leucopus at dietary concentrations up to 25 ppm.

We looked for evidence for the hypothesis that exposure of female birds to polychlorinated biphenyls (PCBs) results in alteration of blood steroid hormone concentrations and alters subsequent hormone transfer of steroids to eggs. Eggs of three-egg clutches were collected from a PCB-exposed common tern (Sterna hirundo) colony (Ram Island, Buzzards Bay, MA, USA) and from a relatively clean colony (Bodkin Island, Chesapeake Bay, MD, USA), and were analyzed for concentrations of organochlorine contaminants and steroid hormones (17beta-estradiol, 5alpha-dihydrotestosterone, testosterone and androstenedione). There was no relationship between total PCBs and steroid concentrations considering all eggs together, considering eggs of different laying order or considering differences between sequentially laid eggs in a clutch. Similarly, concentrations of di- and tri-chlorinated biphenyls and steroids in eggs were not related. The concentrations of PCBs, mercury and selenium were below estimated thresholds for toxicity to embryos. Maternal steroids, except estradiol, were present in yolk of all eggs, with increasing concentrations in the second and third eggs laid. Our data provided no evidence for a maternal toxicological event that might alter the amount of maternal steroid hormone transferred to eggs.

Energy budgets have provided physiological ecologists with a vital link between environmental variables and individual performance and should also prove useful to ecotoxicologists in understanding the effects of sublethal exposure in the field. Exposure to toxic compounds is likely to be metabolically expensive and may result in a trade-off between energy spent to detoxify and excrete contaminants and energy allocated to growth or reproduction. To quantify the energetic cost of polychlorinated biphenyl (PCB) exposure, we fed captive white-footed mice (Peromyscus leucopus) diets containing PCBs (2:1 Aroclor® 1242:1254) at levels of 0, 0.1, 10, and 25 ppm (mg PCBs/kg food). After six weeks on the diets, there were no differences in food intake (g/d), diet digestibility (%), or body mass related to the level of dietary PCBs. This indicated that short-term exposure to PCBs did not cause a detectable increase in energy need as measured by voluntary food intake. We continued to feed mice the PCB-containing diets for one year, at which time we repeated the food intake trial, and also measured oxygen consumption at 20 and 30°C. After one year, all mice had gained mass, but mice on the 25-ppm diet tended to be heavier than mice in the other groups. Compared to the control group, mice on the 25-ppm diet had higher food intake (4.1 vs 3.7 g/d; p = 0.06) and higher oxygen consumption at 30°C (40.1 vs 36.6 ml O2/h; p = 0.01). These results suggest that there is an energetic cost to long-term contaminant exposure that, when combined with other environmental stresses, may influence patterns of energy acquisition and allocation.

Mining and smelting in the Southeast Missouri Lead Mining District has caused widespread contamination of soils with lead (Pb) and other metals. Soils from three study sites sampled in the district contained from approximately 1,000–3,200 mg Pb/kg. Analyses of earthworms [33–4,600 mg Pb/kg dry weight (dw)] collected in the district showed likely high Pb exposure of songbirds preying on soil organisms. Mean tissue Pb concentrations in songbirds collected from the contaminated sites were greater (p < 0.05) than those in songbirds from reference sites by factors of 8 in blood, 13 in liver, and 23 in kidney. Ranges of Pb concentrations in livers (mg Pb/kg dw) were as follows: northern cardinal (Cardinalis cardinalis) = 0.11–3.0 (reference) and 1.3–30 (contaminated) and American robin (Turdus migratorius) = 0.43–8.5 (reference) and 7.6–72 (contaminated). Of 34 adult and juvenile songbirds collected from contaminated sites, 11 (32 %) had hepatic Pb concentrations that were consistent with adverse physiological effects, 3 (9 %) with systemic toxic effects, and 4 (12 %) with life-threatening toxic effects. Acid-fast renal intranuclear inclusion bodies, which are indicative of Pb poisoning, were detected in kidneys of two robins that had the greatest renal Pb concentrations (952 and 1,030 mg/kg dw). Mean activity of the enzyme delta-aminolevulinic acid dehydratase (ALAD) in red blood cells, a well-established bioindicator of Pb poisoning in birds, was decreased by 58–82 % in songbirds from the mining sites. We conclude that habitats within the mining district with soil Pb concentrations of ≥1,000 mg Pb/kg are contaminated to the extent that they are exposing ground-feeding songbirds to toxic concentrations of Pb.

We hypothesized an energetic effect of the pesticide Aldicarb (an acetylcholinesterase inhibitor) through its action on gut innervation, and hence digestive efficiency, and tested this in deer mice (Peromyscus mamculatus Rafinesque) exposed to both the pesticide and cold temperature. Growing deer mice were exposed to Aldicarb in their drinking water at concentrations of 0, 10, and 10,000 ppb and housed at 11 and 23°C. There were no changes due to Aldicarb in several measures of physiological energetics, including energy intake (food consumption, digestive efficiency, gut size) or in the allocation of energy to growth or activity. Two of eight measures of gut size (length and weight of the large intestine) increased significantly with exposure to Aldicarb at the high concentration. Also, there was significant interaction effect of exposure to Aldicarb and cool temperatures on the ratio of mucosal to serosal mass in the small intestine, such that mucosal mass increased when mice were exposed to low doses of Aldicarb at low temperatures but not under other conditions. Although increased absorptive area of the gut (i.e., size or mucosal mass) can be a compensatory response to energy shortfalls, there was no indication of that cause in Aldicarb-exposed mice. Cold-temperature treatment did increase energy costs, however, and both food consumption and gut size increased in mice housed at 11°C, so that digestive efficiency and allocation to growth and activity were no different from those of mice housed at warmer temperatures. Thus, Aldicarb is unlikely to produce any chronic effects on these mice in the wild through disruption of digestion or other causes of energetic shortfall.

The hydrolysis by nuclease P1 of the 16 common deoxydinucleoside monophosphates was examined. The rates of hydrolysis of phosphodiester bond differ by more than two orders of magnitude; dinucleotide monophosphates of the type d(TpN) being most resistant and d(GpN) being next most resistant. The profiles of a mixture of the 16 common dinucleoside monophosphates and of DNA after partial hydrolysis by nuclease P1 and simultaneous treatment with acid phosphatase were compared. The resultant profiles are very similar, except for the appearance of 5-methyldeoxycytidine in the latter. Similar profiles are also obtained from a mixture of dinucleoside monophosphates and from DNA exposed to ionizing radiation beforehand. The 8-hydroxyguanine lesion and a formamido remnant of thymine appear in both profiles as a modified nucleoside and as modified dinucleoside monophosphate respectively. These results suggest that certain radiation induced DNA lesions can be selectively postlabelled based on their resistance to hydrolysis by nuclease P1. The nature of the nuclease P1-substrate interaction is discussed.

American kestrels (Falco sparverius) were fed diets containing methylmercury chloride (MeHg) at 0, 0.6, 1.7, 2.8, 3.9, or 5.0 µg/g (dry wt) starting approximately eight weeks before the onset of egg laying. Dietary treatment was terminated after 12 to 14 weeks, and unhatched eggs were collected for Hg analysis. Blood samples were collected after four weeks of treatment and the termination of the study (i.e., 12–14 weeks of treatment). Clutch size decreased at dietary concentrations above 2.8 µg/g. The average total mercury concentration in clutches of eggs and in the second egg laid (i.e., egg B) increased linearly with dietary concentration. Mercury concentrations in egg B were approximately 25% lower than in the first egg laid and similar in concentration to the third egg laid. Mercury concentrations in whole blood and plasma also increased linearly with dietary concentration. Total Hg concentrations in June blood samples were lower than those in April, despite 8 to 10 weeks of additional dietary exposure to MeHg in the diet. This is likely because of excretion of Hg into growing flight feathers beginning shortly after the start of egg production. The strongest relationships between Hg concentrations in blood and eggs occurred when we used blood samples collected in April before egg laying and feather molt. Environ. Toxicol. Chem. 2010;29:2206–2210. © 2010 SETAC

A physiologically based toxicokinetic (PBTK) model was developed to describe the uptake, distribution, and elimination of methylmercury (CH(3)Hg) in female American kestrels. The model consists of six tissue compartments corresponding to the brain, liver, kidney, gut, red blood cells, and remaining carcass. Additional compartments describe the elimination of CH(3)Hg to eggs and growing feathers. Dietary uptake of CH(3)Hg was modeled as a diffusion-limited process, and the distribution of CH(3)Hg among compartments was assumed to be mediated by the flow of blood plasma. To the extent possible, model parameters were developed using information from American kestrels. Additional parameters were based on measured values for closely related species and allometric relationships for birds. The model was calibrated using data from dietary dosing studies with American kestrels. Good agreement between model simulations and measured CH(3)Hg concentrations in blood and tissues during the loading phase of these studies was obtained by fitting model parameters that control dietary uptake of CH(3)Hg and possible hepatic demethylation. Modeled results tended to underestimate the observed effect of egg production on circulating levels of CH(3)Hg. In general, however, simulations were consistent with observed patterns of CH(3)Hg uptake and elimination in birds, including the dominant role of feather molt. This model could be used to extrapolate CH(3)Hg kinetics from American kestrels to other bird species by appropriate reassignment of parameter values. Alternatively, when combined with a bioenergetics-based description, the model could be used to simulate CH(3)Hg kinetics in a long-term environmental exposure.

Fifty-nine adult male American kestrels (Falco sparverius) were assigned to one of three diet formulations including 0 (control), 0.6, and 3.9 µg/g (dry wt) methylmercury (MeHg). Kestrels received their diets daily for 13 weeks to assess the effects of dietary MeHg on immunocompetence. Immunotoxic endpoints included assessment of cell-mediated immunity (CMI) using the phytohemagglutinin (PHA) skin-swelling assay and primary and secondary antibody-mediated immune responses (IR) via the sheep red blood cell (SRBC) hemagglutination assay. Select hematology and histology parameters were evaluated to corroborate the results of functional assays and to assess immunosuppression of T and B cell-dependent components in spleen tissue. Kestrels in the 0.6 and 3.9 µg/g MeHg groups exhibited suppression of CMI, including lower PHA stimulation indexes (p = 0.019) and a 42 to 45% depletion of T cell-dependent splenic lymphoid tissue (p = 0.006). Kestrels in the 0.6 µg/g group exhibited suppression of the primary IR to SRBCs (p = 0.014). MeHg did not have a noticeable effect on the secondary IR (p = 0.166). Elevation of absolute heterophil counts (p

Changes in the density and distribution of Ixodes dammini, Spielman Clifford, Piesman & Corwin were assessed in Wisconsin by examining hunter-killed deer for ticks in 1981 and 1989. Deer examination sites were distributed widely across the state and included 23 sites in 1981 and 15 sites in 1989; 10 sites were visited both years. Between 1981 and 1989, I. dammini became more abundant throughout its range, and I. dammini range expanded into the southwestern portion of Wisconsin. I. dammini was not found in the eastern one-third of the state. When compared with 1981, the 1989 survey revealed significant increases in the proportion of infested deer in the southwest (Arena), higher levels of infestation of individual deer (number of ticks per individual deer) in the central west (Bangor-W. Salem), and significantly less disparity between the proportions of infested male deer and infested female deer in the north (Spooner) compared with sites farther south. These results are consistent with a pattern of I. dammini range expansion from north to south, followed by an increase in population density in the colonized areas. Dermacentor albipictus Packard was present throughout the range of I. dammini and at sites in the northeastern quadrant of Wisconsin where I. dammini was not found. The range of D. albipictus did not change between the survey years, but its population density increased significantly at sites in the north. There is no evidence for interaction between the two tick species that might affect tick distribution or population density, nor can the greater number of I. dammini found in 1989 be attributed to increased tick activity because of warmer weather; temperatures were cooler in 1989 than 1981.

Several species of wild raptors have been found in Eurasia infected with highly pathogenic avian influenza virus (HPAIV) subtype H5N1. Should HPAIV (H5N1) reach North America in migratory birds, species of raptors are at risk not only from environmental exposure, but also from consuming infected birds and carcasses. In this study we used American kestrels as a representative species of a North American raptor to examine the effects of HPAIV (H5N1) infection in terms of dose response, viral shedding, pathology, and survival. Our data showed that kestrels are highly susceptible to HPAIV (H5N1). All birds typically died or were euthanized due to severe neurologic disease within 4–5 days of inoculation and shed significant amounts of virus both orally and cloacally, regardless of dose administered. The most consistent microscopic lesions were necrosis in the brain and pancreas. This is the first experimental study of HPAIV infection in a North American raptor and highlights the potential risks to birds of prey if HPAIV (H5N1) is introduced into North America.

The purpose of this experiment was to determine the effects of Aroclor 1242, a mixture of polychlorinated biphenyls (PCBs), on plumage characteristics and molt in the American kestrel, Falco sparverius. Several characteristics of plumage, including color and molt schedule, are modulated by hormonal signals and hence may be modified by endocrine-active contaminants. If so, the functions of plumage (e.g., communication for mating or territorial defense) may be compromised by exposure to such compounds. Captive American kestrels were fed Aroclor 1242 at 0, 6.0, and 60.0 ppm (n = 6 males and 6 females per treatment) mixed in their normal diet. Concentrations of plasma estradiol and thyroxine were measured weekly from the beginning of treatment. Measured plumage characteristics included width of the black subterminal band on the tail, color (a composite index of hue and saturation), reflectance from 230 to 800 nm. pattern of feather loss and regrowth on the tail and wing, and timing of onset and duration of molt. Aroclor 1242 depressed plasma thyroxine. Plasma estradiol levels remained low due to the phase of the breeding cycle. Treatments did not disrupt the measured plumage characteristics. This may be due to timing or dose of exposure or to genetic factors.

Polychlorinated biphenyls (PCBs) are environmental contaminants known to cause adverse health effects to biological systems. Limited data are available on their effects on the immune system of wildlife species. Previously, we found that 4 and 6-week-old white-footed mice (Peromyscus leucopus) born from dams injected with a single dose (300 mg/kg) of Aroclor 1254, had altered immunological, hematological, and biochemical responses. Here, we examined the effect of transplacental, lactational and postnatal exposure to Aroclor 1254, at a concentration similar to that found at contaminated sites, on various physiological parameters of 22-week-old white-footed mice. Liver weight and liver somatic index of PCB treated animals were significantly higher, the combined weights of the adrenal glands were significantly lower and EROD and BROD enzyme activity was significantly higher compared to control values. The number of thymocytes of the treated mice was significantly lower than that of the controls; however, thymocytes of treated mice had a higher proliferative response to the mitogen Con A. These alterations were correlated with the PCBs body burdens. Some toxic effects of chronic exposure to PCBs, at levels comparable to exposure found in contaminated sites in the USA, are still evident in adult P. leucopus.

This study examined the density and distribution of the blacklegged tick, Ixodes scapularis Say, at the edge of its range in southern Wisconsin in 1988 and 1989. The study area encompassed sites with ticks at densities ranging from the highest recorded in the state to no ticks detected, as known from previous surveys of white-tailed deer, Odocoileus virginianus Zimmerman, infestation. Ticks were collected from trapped mice, Peromyscus spp., and hunter-killed white-tailed deer. A sharp west-to-east decline in I. scapularis density was identified based on deer infestation rates. At the edge of the ticks range in southern Wisconsin, I. scapularis infestation of Peromyscus Gloger was 1-2 orders of magnitude less than in the middle of the range. In this study, white-tailed deer were more effective than mice for sampling ticks at low density, as is expected given the home range of the hosts and the spatial pattern of ticks. This data set is a baseline for judging changes in the distribution and density of I. scapularis in southern Wisconsin.

The whooping crane is one of the most critically endangered species in North America. The species underwent a severe genetic bottleneck with only 16 individuals remaining in the wild as of 1942. Captive breeding began in 1966 and continues to produce chicks for release in order to establish new wild populations. However, captive birds experience poor reproduction with approximately 40% of eggs being infertile. Males have been known to reach sexual maturity at 5 years of age and continue to reproduce almost as long as the duration of their adult life (i.e. 40 years). Understanding factors affecting seminal quality may assist in identifying and correcting causes of suboptimal reproduction. Our objectives were to determine the influence of age and reproductive seasonality on seminal quality. We hypothesised that seminal quality variations among whooping cranes and ejaculates within a given individual over time were due to bird age and stage of breeding season. In 2010, twenty-nine whooping cranes of 5 age groups housed at Patuxent Wildlife Research Center (Laurel, MD, USA) were studied: ≤5 years (n = 3); 6–10 years (n = 7); 11–15 years (n = 7); 16–20 years (n = 4); >20 years old (n = 8). Semen was collected using a manual manipulation technique at 3 stages of the breeding season: early (March, n = 29) mid (April, n = 24), and late (May, n = 14). Samples were evaluated for seminal volume and sperm concentration, motility, and morphology, with data evaluated by analysis of variance. Bird age had no influence on seminal quality, whereas stage of breeding season affected seminal volume and the proportion of sperm with normal morphology (95% confidence interval). Specifically, samples collected during Mid breeding season had the highest volume (mean ± SEM; early: 42.0 ± 8.0 μL; mid: 66.0 ± 15.2 μL; late: 39.7 ± 17.8 μL), but lowest proportions of structurally normal sperm (early: 78.4 ± 3.7%: mid: 61.5 ± 3.2%; late: 69.7 ± 3.4%). There was a significant difference (P = 0.06) in sperm concentration among stages of the breeding season (early: 66.3 ± 18.8 × 106 sperm mL–1; mid: 179.2 ± 46.2 × 106 sperm mL–1; late: 91.4 ± 47.8 × 106 sperm mL–1). Sperm motility was unaffected by season (early: 36.4 ± 3.5%; mid: 45.9 ± 4.1%; late: 48.0 ± 4.9%). In summary, there is a peak in seminal quality that corresponds with higher volume and more sperm during the mid stage of the season, although with higher instances of structural abnormalities. Despite the small founder base for this species, males in this population produce sperm with no variation in seminal quality across a wide variation in age.

Sixteen wild Peromyscus leucopus, trapped for the establishment of a breeding colony, developed signs of neurological damage (trembling, incoordination, circling, head tilt, and lameness of the rear legs) 2–47 days after capture in southern Wisconsin. Spirochetes were cultured from the brain of 5/11 mice, and Borrelia burgdorferi was cultured from 1 brain. A spirochete was isolated from the bladder of 1 mouse. The spirochete was identified by fluorescent antibody staining with the monoclonal antibody specific for B. burgdorferi, H5332. Serum antibodies to the spirochete were found in 14/15 mice. Negative results were obtained in all tests for viruses and bacteria, including Listeria (2/2), Mycoplasma (2/2), mouse hepatitis virus (10/10), Theilers's encephalomyelitis virus (GD VII) (8/8), REO 3 virus (2/2), and lymphocytic choriomeningitis virus (4/4). There was no bacterial growth from brains cultured on eosin methylene blue or blood agar (3/3). Histologic lesions included nonsuppurative cellular infiltrates in the brain, kidney, liver, and lung. Three outbred Swiss-Webster mice were inoculated orally with a suspension of the brain in BSKII medium, and 3 were inoculated with unpassed B. burgdorferi cultured from the brain of a P. leucopus with motor dysfunction. Five of the inoculated mice developed antibody titers of 1:128; one mouse was positive at 1:256. Motor signs of neurologic damage developed in 3/6 mice 2–24 weeks post-inoculation, and B. burgdorferi was detected in the brains of 2 mice by isolation and by fluorescent antibody.

Sixty breeding pairs of captive American kestrels (Falco sparverius) were exposed to a range of sublethal dietary concentrations of mercury (Hg), in the form of methylmercuric chloride, and their subsequent reproduction was measured. Egg production, incubation performance, and the number and percent of eggs hatched decreased markedly between 3.3 and 4.6 mg/kg dry weight of Hg (1.2 and 1.7 mg/kg wet wt), in the diet. The number of fledglings and the percent of nestlings fledged were reduced markedly at 0.7 mg/kg dry weight (0.3 mg/kg wet wt) and declined further between 2 and 3.3 mg/kg dry weight (0.7 and 1.2 mg/kg wet wt). Dietary concentrations ofor=4.6 mg/kg dry weight (1.7 mg/kg wet wt) were associated with total fledging failure. The estimated decline in fledged young per pair (24%, Bayesian regression) for kestrels consuming 0.7 mg/kg dry weight (0.3 mg/ kg wet wt) raises concerns about population maintenance in areas subject to high inputs of anthropogenic Hg. Mercury concentrations in 20 second-laid eggs collected from all groups were related to dietary concentrations of Hg, and the Hg concentrations in 19 of these eggs were related to eggs laid and young fledged. Concentrations of Hg in eggs from the highest diet group (5.9 mg/kg dry wt; 2.2 mg/kg wet wt) were higher than egg concentrations reported for either wild birds or for captive birds (nonraptors) fed dry commercial food containing 5 mg/kg methylmercury. Accumulation ratios of Hg from diets to eggs were higher than those reported for feeding studies with other species.

We assessed the value of short-tailed shrews (Blarina brevicauda) as a possible biomonitor for polychlorinated biphenyl pollution through measurement of the induction of hepatic cytochrome P450 and associated enzyme activities. First, we checked the inducibility of four monooxygenases (benzyloxyresorufin-O-dealkylase [BROD], ethoxyresorufin-O-dealkylase [EROD], methoxyresorufin-O-dealkylase [MROD], and pentoxyresorufin-O-dealkylase [PROD]) by measuring the activity of these enzymes in hepatic microsomes prepared from shrews injected with beta-naphthoflavone (betaNF) or phenobarbital (PB), typical inducers of cytochrome P4501A (CYP1A) and CYP2B enzyme families, respectively. Enzyme activity was induced in shrews that received betaNF but not in shrews that received PB; PROD was not induced by either exposure. Later, shrews were exposed to a mixture of polychlorinated biphenyls (PCBs) (Aroclor 1242:1254, in 1:2 ratio) at 0.6, 9.6, and 150 ppm in food, for 31 d. Induction in these shrews was measured by specific enzyme activity (BROD, EROD, and MROD) in hepatic microsomes, by western blotting of solubilized microsomes against antibodies to CYP1A or CYP2B, and by duration of sodium pentobarbital-induced sleep. These three CYP enzymes were induced in shrews by PCBs at similar levels of exposure as in cotton rat (Sigmodon hispidus). Neither sleep time nor the amount of CYP2B family protein were affected by PCB exposure. Blarina brevicauda can be a useful biomonitor of PCBs that induce CYP1A, especially in habitats where they are the abundant small mammal.

The purpose of this experiment was to determine the effects of Aroclor 1242, a mixture of polychlorinated biphenyls (PCBs), on plumage characteristics and molt in the American kestrel, Falco sparverius. Several characteristics of plumage, including color and molt schedule, are modulated by hormonal signals and hence may be modified by endocrine-active contaminants. If so, the functions of plumage (e.g., communication for mating or territorial defense) may be compromised by exposure to such compounds. Captive American kestrels were fed Aroclor 1242 at 0, 6.0, and 60.0 ppm (n = 6 males and 6 females per treatment) mixed in their normal diet. Concentrations of plasma estradiol and thyroxine were measured weekly from the beginning of treatment. Measured plumage characteristics included width of the black subterminal band on the tail, color (a composite index of hue and saturation), reflectance from 230 to 800 nm, pattern of feather loss and regrowth on the tail and wing, and timing of onset and duration of molt. Aroclor 1242 depressed plasma thyroxine. Plasma estradiol levels remained low due to the phase of the breeding cycle. Treatments did not disrupt the measured plumage characteristics. This may be due to timing or dose of exposure or to genetic factors.

The effect of the carcinogen acetylaminofluorene (AAF) on nucleic acid structure was examined using 13C- and 31P-NMR spectroscopies. Conformational effects were compared in two AAF-modified dinucleoside monophosphates (ApG and GpA) and two AAF-modified deoxydinucleotides (dpApG and dpGpA). Changes in adenine 13C chemical shifts on formation of the AAF-adduct and as a function of temperature provided evidence of base stacking. Differences in fluorene 13C chemical shifts between the AAF-modified dimer and AAF-modified monomer provided evidence of fluorene stacking. The effect of forming the adduct on the phosphate backbone was examined using 31P-NMR. A correlation was demonstrated between the degree of adenine-fluorene stacking on one hand and the change in conformation of the backbone conformation on the other.

Reactive oxygen radicals have been implicated in diverse pathological processes (1) including the initiation and promotion phases of carcinogenesis (2-4), aging (5), and biological damage following ischemia (6). Two well recognized mechanisms for the production of oxidizing free radicals are via autoxidation processes and by ionizing radiation. It is of interest to compare the products resulting from these two modalities, particularly in relation to damage incurred by DNA constituents. This report describes the products obtained when a DNA model compound, namely 2'-deoxyadenylyl-(3'-5')-thymidine, d(ApT), is exposed to free radicals generated by autoxidizing compounds and by ionizing radiation. Figure 1 a shows the HPLC product profile obtained after d(ApT) was exposed in an oxygenated aqueous solution to ionizing radiation. The major products have been identified previously using NMR spectroscopy and mass spectrometry (7). Figure lb shows the HPLC product profile obtained after d(ApT) was exposed for 48 h at room temperature in an oxygenated aqueous solution to which ascorbic acid was added. Evidently the major products are the same in la and lb. The main product is a formamido modification which probably results from degradation of a precursor 5(6)-hydroxy-6(5)-hydroperoxy-5,6-dihydrothymine modification of d(ApT). Only a formamido fragment of the thymine base remains in this modification which has been described by Cadet et al. (8). Other major products include bases, mononucleotide, 5,6-dihydroxy-5,6-dihydrothymine (glycol) derivatives of d(ApT), and d(ApT) oxidized at the 5' end to give the aldehyde. Not only are the major products generated by the two modalities the same, but the distribution profiles are the same. Analogous results were obtained using 6-hydroxydopamine and dialuric acid as autoxidizing agents. The effect of various mediators on the product profile of d(ApT) obtained via autoxidation was examined to obtain insights into the molecular mechanisms involved. Superoxide dismutase reduced the amount of product somewhat, but the distribution of products appeared unchanged. Catalase, on the other hand, completely inhibited product formation. This result shows clearly that hydrogen peroxide plays a role. It was of interest to determine whether trace amounts of metal complexes could be