Comparison of media to isolate Staphylococcus aureus from teat skin and milking unit liners.

Four procedures were compared for isolation of Staphylococcus aureus from swabbing solutions of teat skin and milking unit liners from commercial dairies. In 2 procedures, 0.1 ml of swabbing solutions were added to either 5 ml Vogel-Johnson or Baird Parker broth media and enriched at 37 degrees C, 4 h. Following enrichment, 0.1 ml culture was transferred to modified Baird-Parker agar and incubated at 37 degrees C, 48 h. In the other 2 procedures, 0.1 ml of swabbing solution was directly placed on either blood or modified Baird-Parker agar plates and incubated at 37 degrees C 48 h. Combining results from all methods, Staphylococcus aureus were isolated from 72 of 913 (7.9%) skin samples, and 34 of 268 liners (12.6%). On average, 43.1% (31/72) of the S. aureus isolates were found by the enrichment in liquid Vogel-Johnson procedure. The average isolation percentage for other methods ranged from 19.4% to 25.0%. Isolation of S. aureus from milking unit liner or teat skin swabbing solutions was approximately twice as likely after enrichment in Vogel-Johnson liquid media as opposed to other methods of isolation. This indicates that enrichment in Vogel-Johnson liquid media improved recovery of S. aureus from swabbing solutions.     

Dual-Energy X-Ray Absorptiometry and Force-Plate Analysis of Gait in Dogs With Healed Femora After Leg-Lengthening Plate Fixation

Dual-energy x-ray absorptiometry was used to measure bone mineral density of four regions in healed femora of nine dogs after fracture fixation with a leg-lengthening plate. Six to 85 months (mean, 46 months) after surgery, the bone mineral density of healed femora was not significantly different from the contralateral uninjured femora ( P >.05; power = 0.8 at Δ= 15%). Radio-lucencies around the proximal screws, apparently associated with screw loosening, were seen on radiographic views of the healed femora of three dogs. In one of these dogs, one screw in the proximal metaphysis had broken. Force-plate analysis of gait was also performed on dogs at the time of bone mineral density measurement. Peak vertical force was decreased in the pelvic limb with the healed fracture compared with the contralateral unoperated limb ( P < 0.05). Clinically apparent lameness in three dogs did not appear to be associated with altered bone mineral density and may have been caused by hip osteoarthritis, a nondisplaced hairline diaphyseal fracture, and screw loosening in conjunction with extensive post-traumatic soft tissue injury.     

Plant interference and chemical toxins in upland pastures

Supernatants derived from desiccated plant material gathered from Agrostis/Festuca vegetation had an inhibitory effect on the germination and early development of seedlings of Trifolium repens cv. S184. Two compounds, o -hydroxyphenylpropionic acid and benzoic acid, were identified chromatographically and their structures confirmed by mass spectrometry. Commercial preparations of the two compounds were effective inhibitors at a concentration of 10⁻² and 10⁻³ m respectively when T. repens was used as the phytometer species. Benzoic acid lost its inhibitory effect when the pH of the two solutions was adjusted to pH 5·5. The roots of seedlings of white clover were severely distorted by o -hydroxyphenylpropionic add at 10⁻³ m . It is likely that phenolic acids, produced from the surface trash created by slot seeding procedures, interfere with the establishment of white clover in upland pastures.     

Severe mortality in broiler chickens associated with Mycoplasma synoviae and Pasteurella gallinarum.

Severe economic loss due to high mortality and condemnation rates occurred on two commercial broiler facilities. Chickens had moderate-to-severe airsacculitis, pericarditis, perihepatitis, tracheitis, and synovitis. Pasteurella gallinarum was isolated from 16 of 18 pericardia, four of 14 livers, 11 of 16 air sacs, six of seven joints and one of 28 tracheas in pure culture. In addition, Mycoplasma synoviae was isolated from trachea and air sac. Lesions were suggestive of an Escherichia coli septicemia, but E. coli was isolated from only four of 28 tracheas and one of 14 livers in pure culture. A coronavirus was isolated from trachea and lung. Whether this coronavirus represented a vaccine or field strain of infectious bronchitis was not determined. These findings suggested that the severe lesions were due to a concomitant infection with an atypical strain of P. gallinarum.     

Transforming growth factor type beta (TGF-beta) and adipogenesis in pigs

The present study was performed on s.c. adipose tissue of fetal pigs at 35 to 110 d of gestation to examine the distribution of TGF-beta-positive cells, to localize TGF-beta immunoreactivity at the cellular level using electron microscopy (EM), and to determine the effect of TGF-beta on primary cultures of pig adipose tissue cells. Tissues for EM were fixed and embedded in LR white resin. Sections then were incubated with a polyclonal antibody specific for TGF-beta and TGF-beta was located using 20 nm colloidal gold conjugated second antibody. Tissues were fixed and embedded in paraffin for localization of TGF-beta at the light microscope (LM) level. Tissues were incubated with anti-TGF-beta followed by localization using biotinylated second antibody. Using LM, only a few cells stained positively for TGF-beta within developing blood vessels at 35 d. By 50 d, more TGF-beta-positive cells were associated with forming capillary networks. Between 70 d and 110 d, positively stained adipocytes usually were clustered around blood vessels. Cells surrounding hair follicles stained positive for TGF-beta between 90 to 110 d. Electron microscopy revealed TGF-beta labeling within fat cells. Fibroblasts and endothelial cells did not exhibit TGF-beta immunoreactivity. The addition of TGF-beta to primary cultures of s.c. adipose tissue cells from newborn pigs prevented lipid filling in fat cells. This effect was dose-dependent, with half-maximal inhibition occurring at 3 pM maximum inhibition occurred at 40 pM. These results indicate that TGF-beta may regulate angiogenic activity and lipid filling in s.c. adipose tissue of fetal pigs. Although TGF-beta was present in adipocytes and in cells associated with developing capillary networks, the physiological role of TGF-beta during early adipose tissue development is not known.     

Morphologic changes in the mammary gland of megestrol acetate-treated and untreated cats: a retrospective study

Abnormal mammary gland growth is a side effect of progestin therapy in some cats. In this retrospective study, the nature and significance of morphologic changes in the mammary gland of 17 megestrol acetate (MA)-treated cats were compared to mammary lesions in 97 untreated cats. Fourteen out of 17 MA-treated cats had non-neoplastic mammary lesions including fibroepithelial hyperplasia (nine cats), lobular hyperplasia (three cats), and duct ectasia (two cats); whereas three MA-treated cats had mammary neoplasms including one adenoma and two carcinomas. Although MA has been causally linked to mammary cancer in cats, only mammary fibroepithelial hyperplasia was clearly associated with MA therapy in this study. Fibroepithelial hyperplasia occurred in older (average age 8.1 years) neutered male and female cats in the MA-treated group and in younger (average age 2.1 years) female cats in the untreated group. Morphologically, both intraductal and solid fibroepithelial growth patterns were seen. Intraductal fibroepithelial hyperplasia was further subdivided into papillary and circumferential types. An apparent greater association between MA therapy and the intraductal types of fibroepithelial hyperplasia was noted. Furthermore, it appears likely that mammary lobular hyperplasia also is linked to MA therapy. Possible mammatrophic effects of MA and other growth-promoting agents in the cat are discussed.     

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.     

Insulin-like growth factor I receptor analysis of satellite cell-derived myotube membranes established from two lines of Targhee rams selected for growth rate

Ovine-derived fibroblasts were used to validate an insulin-like growth factor I (IGF-I) membrane-receptor binding assay system. Competitive binding using fibroblasts revealed that half-maximal inhibition of 125I-IGF-I binding by IGF-I was 2.3 nM. SDS-polyacrylamide gel electrophoresis analysis of specific protein-associated 125I-IGF-I was consistent with the migration of 125I-IGF-I-labeled Type I IGF receptor alpha-subunits at Mr 133,000 daltons. Further, the efficiency of two cell solubilization methods was examined and time-dependent binding equilibrium was determined for the membrane assay system. Satellite cell-derived myotubes were subsequently isolated from primary satellite cell cultures established from the semimembranosus muscles of high and low efficiency-of-gain (EOG) Targhee rams, and IGF-I receptor dynamics were measured. A membrane competitive binding study revealed that half-maximal inhibition of 125I-IGF-I binding was achieved by 1-ng IGF-I for low, and 10-ng IGF-I for high, EOG myotube membrane preparations. Kd values were similar between the high EOG (4.78 nM) and low EOG (2.95 nM) groups; however, receptor concentrations (Bmax) appeared to differ between groups. High EOG membrane receptor Bmax was 3.88 pmole/micrograms protein (19.87 pmole/micrograms DNA), whereas low EOG membrane receptor Bmax was 1.22 pmole/micrograms protein (9.28 pmole/micrograms DNA). These preliminary findings support the hypothesis that genetic selection for EOG results in altered satellite cell responsiveness to IGF-I.     

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).