James A. Render

Effects of waterborne exposure of 17 β-estradiol on secondary sex characteristics and gonads of fathead minnows (Pimephales promelas)

Abstract Environmental contaminants with estrogenic activity have recently received attention because of their potential effects on the reproductive efficiency of humans and wildlife. This study was conducted with the endogenous estrogen, 17 β-estradiol (E2), to establish the histologic response of the fathead minnow (Pimephales promelas) as a model organism. Sexually mature fathead minnows were exposed for 14 days to waterborne concentrations of 1000, 100, 10, 2, 1, 0.5, 0.25, 0.125, 0.1 or 0.0625 nM E2. Exposure to E2 caused a reduction in size of the prominent male secondary sex characteristics, the fatpads and nuptial breeding tubercles. Histological lesions observed in the testes included proliferation of Sertoli cells and degenerative changes. Electron microscopy of seminiferous tubules and their Sertoli cells revealed large phagolysosomes filled with degenerating spermatozoa and other cellular debris. Females had ovaries in which most of the follicles were in the primary stage of development. There were also more atretic follicles and fewer secondary and Graafian follicles than in unexposed females. These findings demonstrate components of sexually mature fish which may be altered by compounds that mimic E2. To determine if lesions observed in males were permanent, 50 sexually mature males and females were exposed to a single concentration of 10 nM E2 for 10 days. Samples were collected from males on the final day of E2 exposure and over a period of 16 weeks after the exposure was stopped. No E2-induced lesions were observed beyond 16 weeks post E2 exposure. Results of these studies suggest that histological lesions could occur at ecologically-relevant exposures to ‘estrogenic’ compounds. However, certain lesions caused by exposure of adult fathead minnows are not permanent.

Clinical and pathologic aspects of spontaneous canine prostate carcinoma: A retrospective analysis of 76 cases

Pet dogs and men share a vulnerability for the development of prostate carcinoma. The purpose of this study was to further characterize the clinical and pathologic features of spontaneous canine prostate carcinoma.

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.

Otitis Media in Preweaned Holstein Dairy Calves in Michigan Due to Mycoplasma Bovis

Mycoplasma bovis was isolated from the tympanic bullae of dairy calves with an exudative otitis media. The history, clinical signs, gross and histologic lesions, and bacteriologic findings are described for 5 preweaned Holstein calves with otitis media from a 600-cow dairy in Michigan. Clinical findings consisted of unilateral or bilateral ear droop, epiphora, head tilt, and recumbency in severely affected calves. Postmortem examination revealed unilateral or bilateral fibrinosuppurative to caseous exudate in the tympanic bullae. Histologically, a marked fibrinosuppurative to caseous exudate filled the tympanic air spaces. The partially ulcerated tympanic mucosa was markedly thickened with mononuclear cell infiltration and proliferation of fibrous connective tissue. Bone remodeling and periosteal hyperostosis were present in some osseous septa. Mycoplasma bovis was isolated from the tympanic bullae of all 5 calves and from the lungs of 2 calves and the frontal sinus of 1 calf. Mycoplasma bovis was isolated at > 100,000 colony forming units/ml from the bulk milk tank of the farm of origin. The isolation of M. bovis from the bulk milk tank, indicating subclinical mycoplasmal mastitis coupled with the feeding of waste milk from mastitic cows to calves is suggestive of a possible source of the infection resulting in otitis media in preweaned dairy calves.

Caprine Mucopolysaccharidosis-IIID: Clinical, Biochemical, Morphological and Immunohistochemical Characteristics

Several animal models have been developed for the mucopolysaccharidoses (MPSs), a group of lysosomal storage disorders caused by lysosomal hydrolase deficiencies that disrupt the catabolism of glycosaminoglycans (GAG). Among the MPS, the MPS-III (Sanfilippo) syndromes lacked an animal counterpart until recently. In this investigation of caprine MPS-IIID, the clinical, biochemical, morphological, and immunohistochemical studies revealed severe and mild phenotypes like those observed in human MPS III syndromes. Both forms of caprine MPS HID result from a nonsense mutation and consequent deficiency of lysosomal N-acetylglucosamine 6-sulfatase (G6S) activity and are associated with tissue storage and urinary excretion of heparan sulfate (HS). Using special stains, immunohistochemistry, and electron microscopy, secondary lysosomes filled with GAG were identified in most tissues from affected goats. Primary neuronal accumulation of HS and the secondary storage of gangliosides were observed in the central nervous system (CNS) of these animals. In addition, morphological changes in the CNS such as neuritic expansions and other neuronal alterations that may have functional significance were also seen. The spectrum of lesions was greater in the severe form of caprine MPS HID and included mild cartilaginous, bony, and corneal lesions. The more pronounced neurological deficits in the severe form were partly related to a greater extent of CNS dysmyelination. These findings demonstrate that caprine MPS HID is a suitable animal model for the investigation of therapeutic strategies for MPS III syndromes.

Proliferation of Maxillary and Mandibular Periodontal Squamous Cells in Mink Fed 3,3′,4,4′,5-Pentachlorobiphenyl (PCB 126)

This report characterizes squamous cell proliferation in young farm mink (Mustela visori) fed a diet supplemented with 0.024 ppm 3,3′,4,4′,5-pentachlorobiphenyl (polychlorinated biphenyl [PCB] congener 126). One to 2 months of dietary exposure to PCB 126 resulted in gross lesions of the upper and lower jaws consisting of mandibular and maxillary nodular proliferation of the gingiva and loose teeth. The maxilla and mandible of the PCB-treated mink were markedly porous because of loss of alveolar bone. Histologically, this osteoporosis was caused by proliferation of squamous cells that formed infiltrating cords. This report clearly documents the fact that the environmental contaminant PCB 126 can cause osteoinvasive squamous proliferation in young mink, although the dose used in the present study was 7 and 36 times higher than what is typically encountered in contaminated bird eggs and fish, respectively.

Dietary Exposure of Mink to Carp from Saginaw Bay, Michigan: 2. Hematology and Liver Pathology

The effects of consumption of environmental contaminants contained in carp (Cyprinus carpio) from Saginaw Bay, Michigan on various hematological parameters and liver integrity of adult female mink (Mustela vison) were determined. Mink were fed diets that contained 0 (control), 10, 20, or 40% carp prior to and throughout the reproductive period (182 days). The diets contained 0.015, 0.72, 1.53, and 2.56 mg polychlorinated biphenyls (PCBs)/kg diet and 1.0, 19, 40, and 81 pg 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs)/g diet, respectively. Mink fed the diets containing carp showed a general dose-dependent occurrence of clinical signs commonly associated with chlorinated hydrocarbon toxicity, including listlessness, nervousness when approached, anorexia, and melena. Erythrocyte counts were less in mink exposed to Saginaw Bay carp than in controls, while the number of white blood cells was greater than in controls. Significant differences (p<0.05) in the concentrations of neutrophils, lymphocytes, monocytes, and eosinophils were also found between the control and carp-fed groups, but are considered to be of limited clinical or biological importance. Hematocrit values for the mink fed the 20 and 40% carp diets were significantly less than those of mink in the control and 10% carp groups. There were no significant differences in hemoglobin concentrations among the groups. Necropsies revealed enlarged yellowish livers in many of the carp-fed mink, especially those fed the 40% carp diet. Liver, spleen, and lung weights of carp-fed mink were significantly greater than those of control mink. Histopathologic examination of the livers revealed various degrees of congestion, hepatocellular fatty changes, and scattered portal lymphocytic infiltration which were most prevalent in mink fed the carp diets. These clinical signs, hematological effects, and histologic alterations are similar to those previously described for chlorinated hydrocarbon toxicoses in mink.

AMYLOPECTINOSIS IN FETAL AND NEONATAL QUARTER HORSES

Three Quarter Horses, a stillborn filly (horse No. 1), a female fetus aborted at approximately 6 months of gestation (horse No. 2), and a 1-month-old colt that had been weak at birth (horse No. 3), had myopathy characterized histologically by large spherical or ovoid inclusions in skeletal and cardiac myofibers. Smaller inclusions were also found in brain and spinal cord and in some cells of all other tissues examined. These inclusions were basophilic, red-purple after staining with periodic acid-Schiff (both before and after digestion with diastase), and moderately dark blue after staining with toluidine blue. The inclusions did not react when stained with Congo red. Staining with iodine ranged from pale blue to black. Their ultrastructural appearance varied from amorphous to somewhat filamentous. On the basis of staining characteristics and diastase resistance, we concluded that these inclusions contained amylopectin. A distinctly different kind of inclusion material was also present in skeletal muscle and tongue of horse Nos. 1 and 3. These inclusions were crystalline with a sharply defined ultrastructural periodicity. The crystals were eosinophilic and very dark blue when stained with toluidine blue but did not stain with iodine. Crystals sometimes occurred freely within the myofibers but more often were encased by deposits of amylopectin. This combination of histologic and ultrastructural features characterizes a previously unreported storage disease in fetal and neonatal Quarter Horses, with findings similar to those of glycogen storage disease type IV. We speculate that a severe inherited loss of glycogen brancher enzyme activity may be responsible for these findings. The relation of amylopectinosis to the death of the foals is unknown.

Ocular and Encephalic Toxoplasmosis in Canaries

In this report we describe the lesions produced by the protozoal organism, Toxoplasma gondii, in the eyes and brain of the common yellow canary (Serinus canaria). Nine of 15 birds in a flock were affected with blindness, which developed over a 3-mo span, and two birds developed torticollis. Microscopic alterations within the eye consisted of a nonsuppurative chorioretinitis with large numbers of macrophages that contained the tachyzoite form of T. gondii in the subretinal space, and aggregates of tachyzoites were found in the nerve fiber layer of the retina with and without necrosis. Tissue cysts with bradyzoites were scattered throughout the meninges and neuropil of the cerebrum and cerebellum. Both forms were confirmed by transmission electron microscopy in the eye and brain. Frozen brain samples reacted with T. gondii-specific cat sera in indirect fluorescent antibody tests. The source of infection was hypothesized to be from a stray cat the owner kept that had access to some of the bird feed. Treatment (trimethoprim 0.08 g/ml H2O and sulfadiazine 0.04 g/ml in water for 2 wk) was instituted by the referring veterinarian on the remaining birds. A second treatment regime was given for 3 wk. The owner of the canaries did not return for further treatment.

Effects induced by feeding organochlorine-contaminated carp from Saginaw Bay, Lake Huron, to laying white leghorn hens. I. Effects on health of adult hens, egg production, and fertility

This study was conducted to determine the effects of consumption of halogenated hydrocarbon compounds, primarily polychlorinated biphenyls (PCBs), contained in Great Lakes fish by the domestic chicken (Gallus domesticus). In this article we report the results of feeding White Leghorn hens for a period of 8 wk diets that contained 31-35% ocean fish and/or carp (Cyprinus carpio) from Saginaw Bay, Lake Huron, MI, which provided 0.3 (control), 0.8 (low-dose group), or 6.6 (high-dose group) mg PCB/kg, wet weight (ww). These concentrations were analogous to 3.3, 26, or 59 pg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalents (TEQs)/g diet, ww, respectively. There were no significant effects on feed consumption among the groups. An unexpectedly high incidence of fatty liver hemorrhagic syndrome (FLHS) was observed in hens from the control (78% FLHS) and low-dose (75% FLHS) groups when compared to the high-dose group (15% FLHS). Birds in the control and low-dose groups had a significant increase in liver and body weights. Significant decreases in egg production, weight, and fertility were immediate in all dose groups, with the effect being permanent in the control and low-dose groups. Although the incidence of FLHS was an unexpected complication, the fact that there were no significant effects on egg production, egg weights, or fertility in the high-dose group suggests that the no-observable-adverse-effect concentration (NOAEC) for these parameters is in excess of 26 mg total weathered PCBs/kg egg, ww. This value was the average concentration of PCBs in the high-dose group eggs during the last week of the study.