A cohort study of the effects of Vietnam service on testicular pathology of U.S. military working dogs.

Using histopathologic diagnoses made on necropsy material from 3,024 military working dogs (MWDs) who died from 1968 to 1973, we analyzed the effect of military service in the Republic of Vietnam on testicular pathology. Among 1,048 MWDs that died in Vietnam and had no reported developmental risk factors for testicular disease, significant excesses of testicular hemorrhage, epididymitis/orchitis, sperm granuloma, testicular degeneration, and seminoma were evident. Among 126 MWDs with prior Vietnam service who died at other duty stations outside Vietnam from 1968 to 1973, significant excesses of testicular degeneration and seminoma were likewise evident. Among 136 MWDs with prior Vietnam service that later died from 1974 to 1980, seminoma continued to be diagnosed in significant excess. In each instance, the odds ratio for the association between Vietnam service and seminoma was 2.0 or greater. Analysis of ever service by Corps Tactical Zones showed significant excesses of seminoma with each Corps area of service compared to Vietnam-era MWDs, but risk was highest in I Corps, particularly at Da Nang Port and Da Nang Air Base. This finding with respect to I Corps is consistent with certain human studies in Vietnam veterans and points to the need for further investigation of possible environmental exposures, particularly those associated to a greater extent with service in I Corps.

U.S. military working dogs with Vietnam service: definition and characteristics of the cohort.

We verified and corrected inaccuracies in descriptive profile information on military working dogs (MWDs) that died from 1965 to 1980 and were reported in the Armed Forces Institute of Pathology Registry of Veterinary Pathology. Using other available military records, we determined which dogs served in Vietnam. Identified were 3,895 MWDs with a unique identity tattoo that served in Vietnam, of which 2,389 served exclusively with U.S. military forces and died in Vietnam. Another 479 Vietnam veteran MWDs completed their service elsewhere. This overall effort resulted in signalment corrections, generally age at death, in 21% of the Registry MWD accessions during the study period. The improved definition and characterization of the Vietnam cohort will lead to greater precision in epidemiologic investigations of the health effects from the Vietnam experience in U.S. military working dogs.

Metastasizing extra-adrenal paraganglioma with neurological signs in four dogs.

Extra-adrenal paragangliomas associated with vertebral pain and clinical neurological abnormalities as a result of metastasis to the vertebral column were diagnosed in four dogs of different breeds by light microscopy. All were males (two intact and two neutered) aged 8 years. Metastatic neoplasms occurred as extradural masses with associated bone lysis at vertebrae C4 (2 cases), T12-L1 (1 case) and L4 (1 case). The neoplastic cells exhibited similar morphology with little variation between cases. All neoplasms showed cytoplasmic granules after staining with the Churukian-Schenk modification of the Pascual argyrophil stain for neurosecretory granules or for membrane bound electron-dense granules (dense-core granules). On immunohistochemical examination the neoplastic cells gave positive results for neuron-specific enolase and negative results for chromogranin and epithelial membrane antigen. Multiple organ metastasis and metastasis to bone have been reported previously, but these cases were unusual due to the involvement of the spine as an apparent predilection site for metastasis, and the sex (male) and age of the animals affected.

Evaluation of Rhipicephalus sanguineus as a potential biologic vector of Ehrlichia platys.

The brown dog tick, Rhipicephalus sanguineus (Acari: Ixodidae), transmits several diseases among dogs including Ehrlichia canis infection. The role of Rhipicephalus sanguineus as a biologic vector for E platys, the rickettsial agent of infectious canine cyclic thrombocytopenia, was studied in dogs. Laboratory-cultured, pathogen-free nymph ticks were fed to repletion on dogs acutely infected with E platys. Tick engorgement coincided with the development of initial parasitemia and thrombocytopenia in the infected dogs. Following repletion, nymph ticks were allowed to molt under controlled conditions. One-month-old E platys-exposed adult ticks failed to infect naive dogs in animal transmission studies. The presence of E platys was not detected in midguts or salivary glands of similarly exposed adult ticks by use of light and transmission electron microscopy. These studies indicate that R sanguineus may not transmit E platys infection.

Brugia pahangi: effects of duration of infection and parasite burden on lymphatic lesion severity, granulomatous hypersensitivity, and immune responses in jirds (Meriones unguiculatus).

The effects of Brugia pahangi infection duration and parasite burden on parasite-associated inflammatory and immune responses were determined over a 181-day period in jirds receiving from one to eight inoculations of infective larvae. Multiple infections did not produce a protective resistance to reinfection as determined by adult worm recovery at necropsy. Intralymphatic granulomatous lesions, lymph thrombi, were first seen at 48 days post initial inoculation (DPI). The numbers of lymph thrombi reached peak levels in singly inoculated jirds at 90 DPI and significantly decreased to low levels by 160 DPI. The ratio of lymph thrombi to adult worms recovered from the spermatic cord lymphatics followed a similar pattern. Sizes of renal lymph nodes, which drain lymphatics containing parasites, followed a temporal pattern of increase and decrease similar to that of lymph thrombi numbers. Peak granuloma areas around antigen-coated beads embolized in lungs were seen at 27 DPI. Granuloma areas around antigen-coated beads began to decrease after 69 DPI and reached sizes not significantly different from uninfected controls by 118 DPI. Multiple inoculations of infective larvae and increasing worm burdens did not affect the pattern of granulomatous response to antigen-coated beads. Eosinophilia of singly and multiply infected jirds peaked at 26 DPI. Eosinophilia of singly infected jirds returned to normal levels by 103 DPI but those of multiply infected jirds remained elevated until 160 DPI. Lymph node cell blastogenic responses to antigen were greater than those of splenocytes at all time intervals measured. However, significant differences in stimulation indexes between groups with different infection durations were not seen with either cell type. Antibody responses to somatic adult worm antigen as measured by ELISA reached near peak levels by 48 DPI and remained elevated for the course of the study in all infected jirds. The decrease in lymphatic lesion severity seen in chronically infected jirds temporally corresponds to the decrease in granulomatous reactivity measured around antigen-coated beads embolized in the lungs. This observation suggests that host and/or parasite factors associated with these two phenomena may be similar. Although these decreases may be the result of down-regulated immune responses, corresponding decreases in antibody levels and blastogenesis of lymphocytes stimulated by crude worm extracts were not observed in chronic infections.

Excess of seminomas observed in Vietnam service U.S. military working dogs.

During the Vietnam War, US military working dogs served with their companion dog handlers in close proximity, sharing common exposures to war-related activity, many zoonotic infectious agents, chemical pesticides, phenoxy herbicides, and extensive use of therapeutic drugs. To gain insight into the effects of the Vietnam experience, we investigated the occurrence of neoplasms in military working dogs based on standard necropsy examination by the Armed Forces Institute of Pathology. We observed that these dogs experienced significant elevated risks for testicular seminoma and, independently, testicular dysfunction. Experimental evidence shows testicular dysfunction and impaired spermatogenesis in laboratory animals exposed to phenoxy herbicides, dioxin, or tetracycline, and antibiotic used extensively in military working dogs in Vietnam. Because an unexplained significant decrease in sperm quality in Vietnam veterans has been observed by the Centers for Disease Control, further research is warranted if we are to clarify military service in Vietnam as a risk factor for testicular dysfunction. The testis should be made a priority site in the study of Vietnam experience-related cancers.

Delayed phalaris grass toxicosis in sheep and cattle.

A phalaris grass (Phalaris caroliniana) caused neurologic signs and lesions in cattle and sheep. The sheep were hyperexcitable and uncoordinated, with severe muscle twitching, stiff gait, and head nodding. The cattle were thin, nervous, and ataxic. One feature of chronic phalaris toxicosis in these cases was onset of signs weeks or months after removal of the animals from the forage.

The morphologic effects of synthetic reproductive steroids on the mammary gland of rhesus monkeys. Mestranol, ethynerone, mestranol-ethynerone, chloroethynyl norgestrel-mestranol, and anagestone acetate-mestranol combinations.

A total of 213 treated and 16 control monkeys comprising 12 experimental groups was evaluated for determination of the long-term (10 years) effects of various dosages of a variety of synthetic oral contraceptive steroids on the mammary glands of rhesus monkeys. The steroid hormones included mestranol, ethynerone, a combination of mestranol and ethynerone, chlorethynyl norgestrel plus mestranol, and anagestone acetate plus mestranol. Various degrees of physiologic lobular hyperplasia and lactational changes were observed in association with all of these steroid hormones; these changes appeared dose-dependent. Mestranol caused a proliferative atypia ranging from a minimal to a moderate degree in 8 of 34 (23%) animals, but it was not dose-related. Eleven of 15 monkeys (73%) administered ethynerone developed proliferative atypia, ranging in degree from minimal to severe, including one invasive carcinoma and 2 lesions resembling intraductal carcinoma in the human. The mestranol and ethynerone combination produced a proliferative atypia in 22 of 52 animals (42%), including five identical to intraductal carcinoma in the human and one identical to lobular neoplasia. Of the 40 monkeys administered anagestone acetate and mestranol, 20 (50%) developed proliferative atypias; the atypias ranged from mild to severe and included five resembling intraductal carcinoma in human breast. The chloroethynyl norgestrel and mestranol combination induced proliferative atypia in 25 of 52 monkeys (49%); six of these atypias were severe and indistinguishable from intraductal carcinoma of the human breast; and one, if in the human breast, would reflect a solid variant of an invasive carcinoma. Only 2 of the 16 control monkeys (12%) developed proliferative atypias, and these were of minimal to mild degree. The occurrence of severe degrees of atypia identical to intraductal carcinoma in the human breast and invasive carcinoma associated with hormone administration suggests a carcinogenic effect.

Ophthalmic corticosteroid therapy: systemic effects in the dog.

Degenerative myopathy suggestive of glucocorticoid-induced myopathy was diagnosed in a 10-year-old female Poodle X Pekingese dog with a history of progressive lameness. Electromyography revealed bizarre high-frequency discharges. Evidence of adrenal suppression was obtained in response to exogenous ACTH. The predominant source of corticosteroid was an ophthalmic preparation used in the management of keratoconjunctivitis sicca. Considering the apparent adverse effects of the corticosteroid in this case, a study was undertaken to determine the systemic effects, if any, induced by long-term ophthalmic administration of 0.1% dexamethasone suspension in healthy adult dogs. Dogs in one group (n = 5) were treated 4 times daily for 8 weeks, and dogs in another group (n = 5) were treated 4 times daily for 16 weeks. All dogs remained healthy throughout the study, but dexamethasone-treated dogs developed marked adrenal suppression, beginning the second week of treatment and intensifying throughout the treatment period. Histopathologic changes in the liver of dexamethasone-treated dogs included scattered foci of vacuolated hepatocytes, increased hepatocytic glycogen content, and ballooning degeneration of hepatocytes. Muscle specimens were histologically, histochemically, and electromyographically normal.

Cerebral phaeohyphomycosis in a dog.

Cerebral phaeohyphomycosis was diagnosed in a 9-year-old spayed dog that had a series of epileptic convulsions a day before death. About 6 weeks before her death, she had been treated for severe demodectic mange. During this period, persistent leukopenia, lymphocytopenia, and thrombocytopenia were found by blood analyses. At necropsy, multiple large pyogranulomatous lesions were found in the cerebrum and meninges. Dematiaceous fungi with brown, branching, septate hyphae and budding yeasts were found within tissue cells and in the necrotic areas.

The morphology of carbon disulfide neurotoxicity.

The morphology of carbon disulfide induced peripheral neuropathy was studied in rats exposed to three concentrations of carbon disulfide by inhalation for 90 days. Rats exposed to 800 ppm developed neurofilamentous axonal swellings in the distal portions of long fibers, including the dorsal ascending sensory and corticospinal tracts of the spinal cord. In peripheral nerve the predominant effect was seen at the level of the posterior tibial nerve. Teased fiber preparations of the muscular branch of the posterior tibial nerve showed numerous paranodal and internodal swellings as well as Wallerian degeneration. Ultrastructurally the swellings were characterized by neurofilament accumulations, decreased numbers of microtubules and thin myelin. Other features included segregation of axoplasmic organelles and cytoskeletal components, intrusion of Schwann cell processes into the axoplasm, Schwann cells with increased cytoplasmic contents, and Schwann cell proliferation around many swollen and demyelinated axons. These features draw important parallels between the morphology of carbon disulfide neuropathy and the neurofilamentous neuropathies induced by hexacarbons and beta,beta' iminodipropionitrile (IDPN).

A 13-week vapor inhalation study of n-hexane in rats with emphasis on neurotoxic effects.

Male and female Fischer 344 rats were exposed to 0-, 3000-, 6500-, or 10,000-ppm n-hexane vapors 6 hr per day, 5 days per week, for 13 weeks. The 13-week exposures had no adverse effect on the growth of female rats. However, the mean body weight gain of male rats in the 10,000-ppm group was significantly lower than for controls at 4 weeks of exposure and thereafter. In addition to the depression of body weight gain, the male exposed to 10,000 ppm had slightly but significantly lower brain weights at necropsy. No adverse testicular effects were noted. Axonopathy was observed in the tibial nerve in four of five male rats from the 10,000-ppm group and one of five male rats in the 6500-ppm group and in the medulla from one male rat in the 10,000-ppm group. These axonal changes were detectable only in teased nerve fiber preparations or in Epon embedded specimens. Histopathologic studies on Formalin fixed tissues did not reveal any lesions that were attributed to n-hexane exposure.

A 90-day vapor inhalation toxicity study of methyl ethyl ketone.

Male and female Fischer 344 rats were exposed to 0, 1250, 2500, or 5000 ppm methyl ethyl ketone (MEK) vapors 6 hours per day, 5 days per week for 90 days. The 90-day exposures had no adverse effect on the clinical health or growth of male or female rats except for a depression of mean body weight in the 5000 ppm exposure group. The 5000 ppm animals had a slight but significant increase in liver weight, liver weight/body weight ratio, and liver weight/brain weight ratio at necropsy. Serum glutamicpyruvic transaminase (SGPT) activity in the 2500 ppm female rats was elevated while the 5000 ppm female rats exhibited significantly decreased SGPT activity. In addition, alkaline phosphatase, potassium and glucose values for the 5000 ppm female rats were increased. Special neuropathological and routine pathological studies did not reveal any lesions that could be attributed to MEK exposure.

Gross and histopathologic effects of medroxyprogesterone acetate and progesterone on the mammary glands of adult beagle bitches.

Adult beagle bitches received 0.2 to 75 mg/kg doses of medroxyprogesterone acetate (MPA) once every 3 months for 1 to 1.5 years. Doses greater than 2.0 mg/kg prevented the recurrence of ovarian cycles; lower doses did not. MPA (10 and 75 mg/kg) induced a dose-related development of multiple, large (greater than 5 mm diameter) mammary nodules that occurred earlier in older dogs than in younger dogs. MPA-induced mammary hyperplasia and nonnodular dysplasias were also noted. Most of the large (5 to 110 mm in diameter) MPA-induced nodules were, in nearly equal numbers, nodules of lobular hyperplasia, simple adenomas, or complex adenomas. Such benign mammopathies all appear to have histopathologic counterparts in human breast disease. However, simple adenomas are uncommon in both humans and dogs. Some of the MPA-induced nodules were benign mixed tumors with cartilaginous metaplasia. No malignant tumors were found. Induction of mammary tumors by 75 mg MPA/kg was not affected by prior ovariectomy but was reached by prior hypophysectomy, suggesting involvement of pituitary secretion in MPA-induced mammary gland disease.

Primary renal tumors in nonhuman primates.

In 17 nonhuman primates, nine females and eight males from 5 to 22 years old, there were 10 cases of renal carcinoma, four of renal adenoma, one nephroblastoma, one hamartoma and one transitional cell papillomatous hyperplasia. The most frequent clinical signs were anorexia, lethargy, weight loss, depression, and dehydration. Tumors were 0.1 to 10.0 cm in diameter. In neoplasms of tubular cell origin, papillary, tubular and solid histologic growth patterns occurred either separately or in combination. Thirty previously reported cases of primary renal tumors in nonhuman primates also were reviewed.

Malignant mammary tumors in beagle dogs dosed with investigational oral contraceptive steroids.

Of 172 beagle dogs administered investigational oral contraceptive steroids for 2.4-5.2 years, 9 developed malignant mammary tumors. At necropsy their ages varied from 41 to 70 months, with a mean age of 4.9 years. The malignant tumors were observed in 1 dog that received ethynerone plus mestranol at 1.05 mg/kg/day and in 4 dogs that received chlorethynyl norgestrel plus mestranol at 1.05 mg/kg/day. Also, 4 dogs that received anagestone acetate plus mestranol at either 0.44 or 1.10 mg/kg/day developed malignant mammary tumors. Malignant tumors were not seen in 33 dogs administered mestranol at 0.02 and 0.05 mg/kg/day for 7 years or in 18 dogs given ethynerone without mestranol at 1.00 mg/kg/day for 5 years. No malignant tumors were observed in 18 control dogs maintained for 7 years without treatment. Three dogs had single malignant mammary nodules, 3 dogs had 2 malignant nodules, 2 dogs had 4-6 malignant nodules, and 1 dog in the treatment group given high dosages of ethynerone plus mestranol had 14 mammary nodules composed of fibrosarcoma. The malignant tumors were histologically classified as 5 anaplastic carcinomas, 2 solid carcinomas, 1 tubular adenocarcinoma, 1 squamous cell carcinoma, and 1 fibrosarcoma. Most dogs had only 1 histologic type of cancer (8/9 dogs); however, 1 dog had carcinomas of both solid and anaplastic types involving different glands. Metastases were present in 5 dogs and most often involved regional lymph nodes and lung.