Fractionation of cock spermatozoa

An account is given of the methodology for fractionation of cock spermatozoa into head and tail fractions by ultrasonication, followed by sucrose density gradient centrifugation. Quantitative estimates of DNA attested to 89.4% purity of the head fraction and low contamination of tails with heads. Recovery of protein and malic dehydrogenase (MDH) activity, following sperm fractionation, averaged 94.3% and 95.7%, respectively. Contamination of the head fraction with tails, as assessed by MDH assay, was only 4.65%, and the purity of the tail fraction was 91%. Intensive succinic dehydrogenase (SDH) activity was histochemically localised in the separated tail fraction and in the tail portion of intact spermatozoa. However, SDH activity was discernible neither in the head fraction nor in the head of intact spermatozoa.     

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.     

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.     

Effect of dietary protein content on episodic growth hormone secretion and on heat production of male broiler chickens.

1. The effect of the crude protein content (200 and 150 g/kg) of isoenergetic diets on episodic growth hormone (GH) release and on heat production was investigated in male broiler chickens. 2. Decreasing the crude protein content of isoenergetic diets from 200 g/kg (HP diet) to 150 g/kg (LP diet) resulted in depressed body weight gain, impaired food conversion efficiency and increased abdominal fat deposition. 3. The pattern of growth hormone secretion was markedly affected by dietary treatment. Broiler chickens fed on the LP diet had higher overall mean, amplitude, baseline and peak frequency than the HP chickens. 4. The LP chickens produced more heat per unit of metabolic body weight than the HP chickens. 5. The hypothesis relating the pattern of GH secretion to protein conversion efficiency was corroborated.     

Distribution of ASFV antigens in pig tissues experimentally infected with two different Spanish virus isolates.

Lymph nodes, spleen, liver, lung and kidney obtained from pigs experimentally infected with two African Swine Fever Virus (ASFV) isolates of differing virulence were fixed by perfusion with glutaraldehyde and embedded in paraffin. An immunoperoxidase technique using a polyclonal anti-ASFV serum was performed on tissue sections in order to detect ASFV antigen. The distribution of ASFV antigen in such infected organs is shown and the differences between both infections compared and discussed. Monocytes, macrophages, hepatocytes, endothelial cells, neutrophils and epithelial cells were found to contain ASFV antigens.     

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.