Measurement of protein turnover in skeletal muscle strips

Isolated porcine and bovine muscle strips were incubated in Krebs Ringer bicarbonate buffer to determine in vitro protein synthesis (PS) and protein degradation (PD) rates to validate the in vitro system for use with livestock species. The addition of 5X plasma concentrations of amino acids to the medium stimulated PS 30%. Addition of 3.5 mM leucine to a leucine-deficient buffer supplemented with amino acids decreased PD 37% and stimulated PS 24%. The addition of .1 U/ml insulin reduced PD 28% and increased PS 30%. Protein degradation was elevated in longitudinally split rat soleus and extensor digitorum longus muscles compared to their contralateral intact muscles. Muscle strips must be removed within 15 min of exsanguination because PD rates become greatly elevated thereafter. ATP concentrations declined during incubation, but the addition of ATP or creatine had no effect on either PD or PS. Neither PD nor PS was affected by the addition of transferrin, fetuin, ascorbate, dexamethasone or indomethacin to the incubation medium. However, muscle strips were sensitive to the addition of triiodothyronine (T3), PD was increased up to 75% as T3 concentration was increased, and PS rates doubled compared to controls. Serum from mature barrows or gilts had no effect on protein turnover, but the addition of 10% and 15% serum from boars increased both PD and PS. With fasted pigs a continual decline in PS occurred over 5 d, whereas PD was elevated at 3 d and then declined to rates comparable to the fed state after 5 d. These data suggest that the in vitro system has application for assessing relative changes that occur in vivo following nutritional, physiological and endocrinological manipulation.     

Secretory patterns and metabolic clearance rate of porcine growth hormone in swine selected for growth

Eleven Landrace pigs (six boars and five gilts, 50 kg) representing lines selected for three generations for maximum weight at 200 d of age were compared to eight pigs (four boars and four gilts, 50 kg) representing contemporary randomly selected Landrace controls to determine the effect of selection for growth on the metabolic clearance rate (MCR) and plasma concentrations of porcine growth hormone (GH). To estimate MCR of GH, the disappearance of a bolus of porcine GH was monitored over 120 min following its i.v. injection. Blood samples also were collected every 15 min over a 6-h period before injecting GH to determine baseline and overall mean GH concentrations, mean peak amplitude and number of GH secretory episodes. Boars exhibited greater overall mean GH concentrations (4.80 vs 3.11 ng/ml; P less than .05) and had greater maximum GH concentrations associated with secretory episodes (16.11 vs 10.80 ng/ml; P less than .05) than did gilts. There were no differences between boars and pigs exhibited greater baseline GH concentrations (2.04 vs 1.25 ng/ml; P less than .01) than did those from the unselected Landrace line. Selected and control pigs exhibited similar (P greater than .15) overall mean concentrations of GH, frequency of secretory episodes, amplitude of GH peaks and MCR. These data demonstrate that pigs selected for heavier weight at 200 d of age had greater basal plasma GH concentrations than did unselected control pigs.     

Immunological cross-reactivity between pseudocapsular antigens of strains of Staphylococcus aureus isolated from cases of bovine mastitis

An assay was developed to measure the degree of immunological cross-reactivity between the pseudocapsules of a vaccine (reference) strain of Staphylococcus aureus and pseudocapsules of strains of S. aureus isolated from cases of bovine mastitis. The field strains were obtained from Australia, New Zealand, Norway, the U.K. and the U.S.A. There was large variation among strains in cross-reactivity of their pseudocapsules with those of the reference strain. For 104 Australian strains, the range of cross-reactivity indices (CRI) was 6.1-63.2% (on a scale of 0-100%, with 0% being complete identity and 100% being nil identity); for 61 overseas strains the range of CRI values was 25.7-72.1%. The data indicated that pseudocapsule antigens of Australian strains were antigenically more closely related to those of the reference strain than were pseudocapsular antigens of strains from the 4 other countries.     

Adhesion of Staphylococcus aureus and Escherichia coli to bovine udder epithelial cells

An assay for the adhesion of tritiated thymidine-labelled Staphylococcus aureus and Escherichia coli to bovine mammary ductular epithelial cell lines was developed. The relative adhesion of 15 strains of S. aureus to these cell lines was examined. Four strains did not adhere and the remaining 11 adhered at variable levels. Adhesion to different cell lines was generally similar. Adhesion to freshly collected bovine mammary epithelial cells was significantly greater than that to cells maintained in tissue culture. The system described was demonstrated to be a suitable model for studying adhesion of mastitis-causing organisms to bovine mammary epithelial cells.     

Influence of dietary protein on the pathophysiology of haemonchosis in lambs given continuous infections

Mixed-sex groups of three-month-old Finn Dorset and Dorset Horn lambs were given complete diets containing either high or low protein. A trickle infection with Haemonchus contortus of 200 larvae, given three times a week, began one month later for a 17 week period. Erythrokinetic, metabolic, digestibility and balance studies were conducted with the male lambs during weeks 12 to 14 of the infection. Clinical signs of haemonchosis became evident only in the low protein diet group. The infection was characterised by severe macrocytic anaemia, hypoproteinaemia, hypoalbuminaemia, reduced survival, high egg counts and a significantly greater worm burden. In the high protein diet group many lambs developed resistance to further infection in contrast to the low protein group in which no resistance developed. Live-weight gain was not affected by infection in the high protein diet group but decreased in the lambs given the low protein diet compared with uninfected controls. However, infection did not appear adversely to affect the digestive efficiency, nitrogen retention or iron absorption in either dietary group although loss of appetite occurred in the low protein group. Examination of a sample joint from the carcases showed infection was associated with a marked reduction in muscle mass in lambs given the low protein diet relative to the control animals.     

Differential effect of T-2 toxin on murine host resistance to three facultative intracellular bacterial pathogens: Listeria monocytogenes, Salmonella typhimurium, and Mycobacterium bovis

The effect of T-2 toxin, a radiomimetic immunosuppressive agent, on resistance to the facultative intracellular bacterial pathogens Listeria monocytogenes (strain EGD), Mycobacterium bovis (BCG Copenhagen 1331), and Salmonella typhimurium was determined. Female Swiss ICR mice were given a single dose of T-2 toxin (4 mg/kg of body weight) by gastric gavage. On the seventh day after toxin administration, the mice were infected by intraperitoneal inoculation with L monocytogenes, S typhimurium, or M bovis. Mice given the toxin also were exposed to respirable droplet nuclei containing L monocytogenes or M bovis. The effect of the toxin on the course of infection was monitored by observing mortality or by enumeration of bacteria in the spleen or lungs of infected mice. The toxin increased resistance to infection with L monocytogenes initiated by intraperitoneal inoculation, but reduced resistance to M bovis infection initiated by intraperitoneal inoculation. The toxin had no appreciable effect on the course of salmonellosis or on resistance to infection initiated by inhalation of L monocytogenes or M bovis aerosols. Therefore, it was concluded that T-2 toxin does not necessarily reduce resistance to infection in mice. The toxin's effect on the course of in vivo bacterial infections depends on the nature of the infective agent and the route of inoculation.