Pathogenesis of placentitis in the goat inoculated with Brucella abortus. II. Ultrastructural studies

Pregnant goats were inoculated intravenously or in uterine arteries with Brucella abortus, and tissues from the uterus and placenta were examined by electron microscopy. Identification of B. abortus in placentae was with antibody-coated colloidal gold. B. abortus was first seen in phagosomes of erythrophagocytic trophoblasts and in the rough endoplasmic reticulum of chorioallantoic trophoblasts. Subsequently, trophoblast necrosis and ulceration of chorioallantoic membranes were present. Coincidently, B. abortus was present in the lumen of placental capillaries. In late stages of infection, placental vasculitis was present, and placentomal trophoblasts were separated from maternal syncytial epithelium. In lesions with vasculitis, large numbers of B. abortus were in connective tissue of chorionic villi. Within the placentome, trophoblasts that lined chorionic villi contained no intracellular bacteria and were separated from B. abortus by intact basement membranes. These results suggest that bacteremic B. abortus is endocytosed by erythrophagocytic trophoblasts and that B. abortus replicates in the rough endoplasmic reticulum of chorioallantoic trophoblasts. Replication of brucellae in trophoblastic rough endoplasmic reticulum is unique; we believe that B. abortus may utilize endoplasmic reticulum for synthesis and glycosylation of bacterial membrane proteins or that B. abortus catabolizes trophoblast secretory proteins.     

Outcome of limb amputations in wapiti: 13 cases (1995-2001).

The object of this investigation was to determine the outcome of limb amputation in wapiti. Medical records of 13 wapiti that underwent limb amputation were reviewed to determine age, weight, sex, injury preceding amputation, limb amputated, amputation location, length of hospital stay, complications, and outcome. Ages ranged from 3 months to 8 years and weights from 70 kg to 280 kg. All animals were female. Eleven animals sustained catastrophic long bone fractures prior to amputation. Five animals had front limb amputations and 8 had hind limb amputations. Of these animals, 1 with a front limb amputaiton and 5 with hind limb amputations survived. Postsurgical inability to stand is associated with hospitalization of > 1 day. Of the 13 wapiti reviewed, 5 with hind limb amputations have produced and reared offspring. Hind limb amputation can be considered a viable alternative to euthanasia when catastrophic orthopedic injuries occur in female wapiti.     

Epidemiology and diagnosis of Mycobacterium tuberculosis in captive Asian elephants (Elephas maximus).

The deaths of two Asian elephants (Elephas maximus) in August 1996 led the United States Department of Agriculture to require the testing and treatment of elephants for tuberculosis. From August 1996 to September 1999. Mycobacterium tuberculosis infection was confirmed by culture in 12 of 118 elephants in six herds. Eight diagnoses were made antemortem on the basis of isolation of M. tuberculosis by culture of trunk wash samples; the remainder (including the initial two) were diagnosed postmortem. We present the case histories, epidemiologic characteristics, diagnostic test results, and therapeutic plans from these six herds. The intradermal tuberculin test, enzyme-linked immunosorbent assay serology, the blood tuberculosis test, and nucleic acid amplification and culture are compared as methods to diagnose M. tuberculosis infection in elephants.     

Chemopreventive efficacy of lycopene on 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis.

The chemopreventive efficacy of lycopene on 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis was examined using lipid peroxidation, reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR) as biomarkers of chemoprevention. Twenty four male Syrian hamsters were divided into four groups of six animals each. The right buccal pouches of the animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin three times a week. The animals in group 2 were painted with DMBA as in group 1 and in addition received 2.5 mg/kg body weight lycopene orally three times a week on days alternate to DMBA application. Group 3 animals received lycopene as in group 2. Animals in group 4 received neither DMBA nor lycopene and served as control. The hamsters were killed after an experimental period of 14 weeks. Biochemical measurements were carried out in tumour and normal tissues. All hamsters painted with DMBA alone for 14 weeks developed well-differentiated squamous cell carcinomas. Diminished lipid peroxidation in the oral tumour tissue was accompanied by a significant increase in the levels of GSH, GPx, GST and GR. Administration of lycopene significantly suppressed DMBA-induced oral carcinogenesis as revealed by the absence of carcinomas. The results of the present study suggest that lycopene may exert its chemopreventive effects by modulating lipid peroxidation and enhancing the activities of the enzymes in the glutathione redox cycle.     

P‐20 Clinical,cytopathological and histopathological evaluation of sporotrichosis in experimentally infected cats

The purpose of this study was to evaluate the clinicopathological aspects of experimental sporotrichosis in cats and compare the sensitivity of cytopathology, histopathology and culture as diagnostic tools in different phases of the infection. Twenty adult, mixed‐breed cats (10 males and 10 females) were inoculated subcutaneously with 10⁶ fungal microorganisms. Clinical examination was performed weekly. Cytopathologic, histopathologic and culture examinations were performed at 15, 30 and 60 days postinoculation. Culture of multiple organs was performed after euthanasia at 30 (10 cats) and 60 (10 cats) days postinoculation. Friedman parametric and nonparametric statistical analysis were applied to the results. The nodular, tumoral and necrotic lesionsprogressed significantly until day 30 postinoculation, and partial spontaneous regression occurred at day 60, particularly in males. An intense inflammatory pyogranulatomous and lymphocytic infiltrate with rare giant cells and sparse fibrosis associated with numerous, pleomorphic, intra‐ and extracellular fungal cells were observed on day 30. These findings gradually decreased by day 60. Despite the inflammatory granuloma associated with feline sporotrichosis, a tendency for dissemination was observed, with fungal isolation in the lymph nodes, spleen and liver at the 30 and 60 days postinoculation. No significant differences were observed between cytopathology, histopathology and fungal culture during the different phases of the disease. Therefore, cytological examination was considered a simple, rapid and inexpensive diagnostic method at all stages of this disease. Funding: Self‐funded.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.