Isolation of Mycobacterium paratuberculosis after oral inoculation in uninfected cattle.

Feces from cows naturally infected with Mycobacterium paratuberculosis was given to 6 uninfected heifers by orogastric intubation, to determine whether ingested organisms could be passively excreted and detected by bacteriologic culture of feces (ie, false-positive result). Heifers were paired, and each pair received a different dose of feces on days 1 and 2. Fecal samples were collected from the heifers 3 times daily. Mycobacterium paratuberculosis was detected in fecal samples of all heifers within 18 hours of being given the first dose of feces. The number of colony-forming units peaked on days 3 or 4, and organisms were no longer detected by day 7. The number of colony-forming units in fecal samples from the heifers was approximately proportional to the dose given. On days 15 and 16, the experiment was repeated with feces from a second infected cow. Results were similar to those in the first experiment. All heifers remained seronegative (agar-gel immunodiffusion test and ELISA) and had negative results to the intradermal johnin test throughout the experiment. Lymph node and intestinal tissues were obtained from all 6 heifers at slaughter on day 28. Mycobacterium paratuberculosis was not isolated from mesenteric lymph nodes from the ileocecal valve region, but was isolated from ileal mucosal samples from each heifer.     

Ionized calcium concentration in horses with surgically managed gastrointestinal disease: 147 cases (1988-1990).

Packed cell volume, total plasma protein, serum sodium, potassium, and ionized Ca2+ concentrations, and blood pH were determined at the time of admission and following surgery in 147 horses with acute abdominal crisis. Horses were allotted to 3 categories on the basis of the surgical lesion: (1) nonstrangulating obstruction of the ascending or descending colon (category A, n = 76), (2) strangulating and nonstrangulating infarction of the cecum or ascending colon (category B, n = 37), and (3) strangulating and nonstrangulating infarction of the small intestine (category C, n = 25). Horses with low serum ionized Ca2+ concentration following surgery were given 23% calcium gluconate (100 to 300 ml) IV to effect, and ionized Ca2+ concentration was determined following treatment. The serum ionized Ca2+ concentrations of horses in categories A, B, and C before and after surgery were lower than our normal laboratory reference range. Prior to surgery, serum ionized Ca2+ concentration measured from horses in category B and C was lower than that in horses in category A. There was no difference in ionized Ca2+ concentration in serum samples obtained before surgery in horses from category B and C, and in serum samples obtained following surgery. There was a decrease in ionized Ca2+ concentration during surgery in horses in category A. There was no change between preoperative and postoperative ionized Ca2+ concentration in the samples obtained from horses in category B and C. After calcium gluconate administration, all horses with low serum ionized Ca2+ after surgery had concentrations within our normal range. Measurement of serum ionized Ca2+ in horses with an acute abdominal crisis is recommended.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of Tissue Adhesive to Contain Axonal Regeneration in Horses

Bilateral palmar and plantar digital neurectomies were completed in 10 horses (a total of 80 neurectomies) using one of three methods: (1) simple transection (guillotine method); (2) epineural capping; (3) n-butyl cyanoacrylate injected into the epineural sheath to act as a nerve sealant. Horses were regularly evaluated clinically for tenderness in and around the surgical site, as well as skin sensation at the coronary band in the heel region, during the 12-week course of the study. None of the surgical sites exhibited any signs of drainage or infection. Horses were then euthanatized, the nerve stumps were dissected from surrounding tissues, and the length and width of the tissue mass that had formed on the end of the nerve was recorded. Longitudinal and transverse sections of the nerve endings were examined histologically for numbers of proliferating axon sprouts (neuroma formation); whether the axons had penetrated the epineurium; degree of Schwann cell proliferation; degree of chronic inflammation; extent of foreign body reaction; extent of retrograde degeneration of the nerve bundles; and amount of fibro vascular proliferation. The proportion of legs exhibiting tenderness or heel sensation did not differ significantly between the three different treatments at any of the six different times they were examined. There was no difference between the three treatments in the length or width of the fibrous tissue scar on the ends of the nerves or in the number of sprouting axons from the ends of the nerves. Of 80 nerves examined, only two nerves were not confined to the epineurium. Both these nerves had been treated by simple transection. Statistically there was more chronic inflammation and foreign body reaction in the acrylic treated nerves, but no difference in Schwann cell proliferation or retrograde degeneration between the three treatments. There was slightly less fibrovascular proliferation in the transected nerves than in those subjected to epineural capping or acrylic, but the difference was not statistically significant. The use of the tissue adhesive n-butyl cyanoacrylate to prevent the continuous growth of axons after digital neurectomy seems to offer little advantage over more traditional methods of neurectomy.     

Oxytocin synthesis and secretion from bovine corpora lutea exposed in vitro to cycloheximide and colchicine

Two experiments were conducted to study the effects of cycloheximide and colchicine on prostaglandin F₂ (PGF₂)-induced secretion and synthesis of oxytocin in bovine luteal tissue in vitro. Corpora lutea were collected from beef heifers on Day 8 of the estrous cycle. In Experiment 1, incorporation of [¹⁴C]-leucine into oxytocin synthesized and secreted by luteal slices after exposure to PGF₂, cycloheximide and cycloheximide plus PGF₂ was examined. In Experiment 2, synthesis and secretion of oxytocin were evaluated in luteal slices incubated with colchicine and PGF₂ alone and in combination. Cycloheximide inhibited incorporation of labeled leucine into luteal proteins by more than 90% and no labeled oxytocin was detected in the media or tissue. Prostaglandin F₂ induced significant secretion of oxytocin that was not inhibited by cycloheximide. Tissue levels of oxytocin after incubation with cycloheximide and/or PGF₂ did not differ and were similar to those of the incubated control. Colchicine alone did not suppress oxytocin secretion and did not alter the ability of PGF₂ to induce significant secretion of this nonapeptide. Tissue concentrations of oxytocin after incubation with colchicine and/or PGF₂ did not differ. These studies indicate that secretion and replenishment of luteal oxytocin in vitro is not contingent upon de novo protein synthesis. Inability of colchicine to suppress oxytocin secretion and synthesis may have been due to the short duration of exposure of luteal tissue to the drug.     

Hyperadrenocorticism in four cats.

This paper describes four cats with hyperadrenocorticism. Cat 1 showed polydipsia and polyphagia. Diabetes mellitus was initially diagnosed. As the animal appeared to be insulin resistant, pituitary and adrenocortical function tests were performed and the diagnosis of hyperadrenocorticism was made. Resistance to the high-dose dexamethasone suppression test was noticed in this cat. Pathological examination revealed a pituitary chromophobe adenoma. Cat 2 presented with diabetes mellitus, which was treated with insulin. The animal had a pendulous abdomen and its coat was in a poor condition. The low-dose dexamethasone suppression test demonstrated hyperadrenocorticism. Necropsy findings of pituitary tumour and hyperplasia of the adrenal cortex confirmed the diagnosis. Cat 3 showed clinical abnormalities indicative of hyperadrenocorticism, for instance, muscle weakness, alopecia, multiple abscesses. The diagnosis of hyperadrenocorticism was confirmed by the results of the lowe-dose dexamethasone suppression test. Pathological examination revealed an adrenocortical carcinoma. Cat 4 presented with polydipsia. The cause of this symptom was not found initially. One and a half years later additional symptoms, such as nephritis and polyphagia developed. Hyperadrenocorticism was diagnosed because of a palpable mass cranial to the left kidney. The diagnosis was confirmed by the results of the lowe-dose dexamethasone suppression test and the necropsy findings.     

Pathology of infectious diarrhea of infant rats (IDIR) induced by an antigenically distinct rotavirus

Suckling rats were inoculated with a group B rotavirus to determine the progression of the morphologic changes induced in the intestine by this virus. Several changes were observed by light microscopy 1 day after viral inoculation: shortening of small intestinal villi, villous epithelial necrosis, and villous epithelial syncytia. The lesions were most often present in the distal small intestine, although other small intestinal segments were affected to a lesser degree. By day 3 post-inoculation, epithelial necrosis, and syncytia were no longer present; however, the villous epithelium was disorganized and irregularly vacuolated, and intestinal crypt epithelium was hyperplastic. Alterations in villous height to crypt depth ratios were present in portions of the small intestine for the remainder of the 12-day study period. Epithelial syncytia appeared to form by the breakdown of the lateral interdigitating membranes of the absorptive villous epithelium. Viral particles, abundant in the syncytia, appeared to form from amorphous or reticular arrays of viral precursor material. Group B rotaviral antigens, as detected by indirect immunofluorescence, were present in large amounts in the small intestinal villous epithelium only on the first day after viral inoculation. These studies show that two important diagnostic features of group B rotaviral infections of rats, epithelial syncytia and viral antigen as determined by immunofluorescence, are present only on the first day of disease. These findings should be taken into consideration when attempting to diagnose disease induced by this agent.     

Correlation of DNA ploidy to tumor histologic grade, clinical variables, and survival in dogs with mast cell tumors.

By using flow cytometry, a retrospective analysis of the DNA content of 40 primary canine mast cell tumors and seven lymph nodes that contained metastatic mast cell tumor from 44 dogs of various breed, sex, and age was performed on formalin-fixed, paraffin-embedded samples of the tumors and nodes. These samples were chosen according to the following criteria: samples contained sufficient well-preserved tumor tissue in the paraffin block for processing, sufficient patient history data were available, clean and homogeneous cell suspensions were obtained after processing, and interpretable DNA histograms were produced on analysis. The ploidy data obtained were compared with the histopathologic grade, the anatomical site of occurrence, the clinical stage of the tumors, and the survival of the dogs. Over 70% (29/40) of the mast cell tumors were diploid. Three metastatic mast cell tumors in lymph nodes had the same ploidy status as their corresponding primary tumors. In five dogs, mast cell tumors from multiple sites in each dog displayed similar ploidy status. Of 26 dogs evaluated for survival times, 69% (18/26) had diploid tumors and 31% (8/26) had aneuploid tumors. When numbers of diploid versus aneuploid tumors were compared, no significant difference was found between any two grades, clinical stages, or anatomic sites. A significant difference (P = 0.02) was found, however, between aneuploid and diploid tumors when comparing Stage I and non-Stage I disease. The Kaplan-Meier survival plot indicated a tendency towards an increased survival within the first year in dogs with diploid versus aneuploid tumors (P = 0.06).