Experimental staphylococcal mastitis in the mouse: the persistence of chronic infection from one lactation to the next.

Chronic staphylococcal mastitis was established in 50 mammary glands of 25 mice. The mice were mated and in the subsequent lactation there was alveolar mastitis in one of 50 glands. A further five glands had abscesses that contained staphylococci but there was no alveolar mastitis. The results are discussed in relation to the large number of infections that persist from one lactation to the next in cows not subjected to mastitis control measures.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 5. Kinematic effects

AIM: To examine the effect of race training on kinematic parameters of the trot in a sample of 2-year-old Thoroughbred fillies. METHODS: Two-year-old Thoroughbred fillies that had been either trained for racing (n=7) or left untrained in small paddocks (n=7) were examined for lameness, and linear and temporal kinematic data were collected from video recordings of each horse trotted in-hand, 5, 9 and 13 weeks after the beginning of training. Horses were graded as: sound (Grade 0), suspected but not obviously lame (Grade 1), or definitely lame at the trot (Grade 2). RESULTS: Mean velocity at the trot did not vary with lameness grade. Compared with horses classified as sound (Grade 0), horses classified as suspected or definitely lame (Grades 1 and 2) had shorter (2.55 (SE 0.02) vs 2.41 (SE 0.02) and 2.44 (SE 0.03) m; p<0.001) and faster (671 (SE 3) vs 645 (SE 1) and 647 (SE 4) msec; p<0.001) strides. None of the linear or temporal parameters measured varied significantly between the successive observation periods. When data from sound horses were pooled, the training group trotted at a higher mean velocity (4.22 (SE 0.08) vs 3.22 (SE 0.06) m/sec; p<0.001) and with a longer stride length (2.79 (SE 0.05) vs 2.18 (SE 0.06) m; p<0.001) than the untrained group. The stride duration was longer (669 (SE 7) vs 662 (SE 9) msec; p=0.01), stance period was shorter (34.1 (SE 0.6) vs 39.3 (SE 0.7) %; p<0.001) and mid-stance was achieved earlier in the stride (12.1 (SE 0.6) vs 13.1 (SE 0.2) %; p=0.001) in the trained than the untrained group. A longer swing phase in the trained group was associated with an increase in retraction time (9.2 (SE 0.2) vs 7.6 (SE 0.3) %; p=0.001). CONCLUSIONS: These data indicate that early race training in young Thoroughbreds was associated with quantifiable changes in linear and temporal kinematic parameters of the trot, which were related to the race-training objective of improving the horse's ability to work at higher velocities. CLINICAL RELEVANCE: Race training resulted in changes in the gait of non-lame horses at the trot, which should be considered during veterinary lameness examinations.     

Uterine mast cells and immunoglobulin-E antibody responses during clearance of Tritrichomonas foetus

We showed earlier that Tritrichomonas foetus-specific bovine immunoglobulin (Ig)G1 and IgA antibodies in uterine and vaginal secretions are correlated with clearance of this sexually transmitted infection. Eosinophils have been noted in previous studies of bovine trichomoniasis but the role of mast cells and IgE responses have not been reported. The hypothesis that IgE and mast cell degranulation play a role in clearance was tested in 25 virgin heifers inseminated experimentally and infected intravaginally with T. foetus strain D1 at estrus and cultured weekly. Groups were euthanatized at 3, 6, 9, or 12 weeks, when tissues were fixed and secretions were collected for culture and antibody analysis. Immunohistochemistry using a monoclonal antibody to a soluble lipophosphoglycan (LPG)-containing surface antigen (TF1.17) demonstrated antigen uptake by uterine epithelial cells. Lymphoid nodules were detected below antigen-positive epithelium. Little IgG2 antibody was detected but IgG1, IgA, IgM, and IgE T. foetus-specific antibodies increased in uterine secretions at weeks 6 and 9 after infection. This was inversely proportional to subepithelial mast cells numbers and most animals cleared the infection by the sampling time after the lowest mast cell count. Furthermore, soluble antigen was found in uterine epithelium above inductive sites (lymphoid nodules). Cross-linking of IgE on mast cells by antigen and perhaps LPG triggering appears to have resulted in degranulation. Released cytokines may account for production of predominantly Th2 (IgG1 and IgE) and IgA antibody responses, which are related to clearance of the infection.     

Evaluation of a commercially available enzyme-linked immunosorbent assay for detecting antibodies to Fasciola hepatica and Fasciola gigantica in cattle, sheep and buffaloes in Australia

A commercially available ELISA for detecting antibodies to liver fluke was evaluated for use in Australia. Milk and serum samples from cattle and sheep in which infection with Fasciola hepatica was confirmed by detection of eggs in faeces were used to estimate sensitivity. Similar samples collected from cattle and sheep outside the F. hepatica-endemic area were used to estimate specificity. The ELISA was also evaluated for detecting antibodies to F. hepatica in milk from sheep and antibodies to Fasciola gigantica in sera from cattle and buffaloes, but with small numbers of samples. In cattle, the sensitivity and specificity of the ELISA were 98.2% and 98.3% using serum and 97.7% and 99.3% using milk. In infected herds, 41.4% and 41.5% of animals were positive in the serum and milk ELISAs, respectively, whereas F. hepatica eggs were found in faecal samples from 26.5% of animals. In sheep, the sensitivity of the ELISA was 96.9% and the specificity was 99.4%. In infected flocks, 60.2% of animals were positive in the serum ELISA and F. hepatica eggs were found in faecal samples 52.2% of animals. There was perfect agreement in the ELISA between paired serum and milk samples collected from ewes. The assay detected antibodies in sera from cattle and buffaloes with natural and experimental F. gigantica infections. In the experimentally infected animals, antibodies were detected 2 weeks post-infection. We conclude that the ELISA will be a valuable tool for diagnosing F. hepatica infections in cattle and sheep. The assay may also be useful for diagnosing F. gigantica infections but further studies are required to establish sensitivity and specificity.     

The carboxyterminal processing protease of D1 protein: expression, purification and enzymology of the recombinant and native spinach proteins

The carboxyterminal processing protease of D1 protein (CtpA) is predicted to be an excellent target for a general broad-spectrum herbicide. The gene for spinach CtpA has been expressed in Escherichia coli. The expressed protein that was found mainly in inclusion bodies has been purified and refolded on a nickel-chelate column. Active recombinant CtpA was recovered. Two assays for CtpA activity were developed, a medium-throughput HPLC assay using a fluorescent substrate and a high-throughput assay based on fluorescence polarization capable of application in a high-throughput 96-well plate format. This high-throughput assay was developed to screen chemistry for CtpA inhibitors. Native spinach CtpA was partially purified and the native and recombinant enzymes were compared kinetically for their K(m) and V(max) values using different peptide substrates. Native CtpA partially purified from spinach was shown to have similar kinetic properties to recombinant CtpA. Antibodies developed against the recombinant protein were used to estimate the in planta abundance of the native enzyme in spinach. Since only a small proportion of the recombinant protein is refolded during isolation and it appears that only a small proportion of this enzyme is active, size-exclusion chromatography and light scattering experiments were performed on rCtpA in order to gain insight into its structure and the reasons why most of the protein is not active. The use of rCtpA to screen for herbicidal compounds and the more general question of how good a herbicide target the enzyme is are discussed.     

The experimental production of krimpsiekte in sheep with Tylecodon grandiflorus (Burm.f.) Toelken and some of its bufadienolides

Six bufadienolides were isolated from Tylecodon grandiflorus ( Burm .f.) Toelken . The paretic syndrome, krimpsiekte , could be induced in sheep either by repeated oral administration of small quantities of plant material or the intravenous injection of small quantities of certain bufadienolides . A mild to moderate, acute to subacute, multifocal cardiomyopathy was evident in sheep poisoned by both the plant and the bufadienolides .     

Changes in plasma enzyme activities and other blood components in response to acute and chronic liver damage in cattle

Three calves were dosed orally with carbon tetrachloride at 0.05, 0.1 or 0.3 ml/kg body-weight. Plasma enzyme activities, clotting times, bilirubin concentrations and bromsulphthalein elimination times increased. These changes are associated with damage to the liver and are discussed in relation to the clinical syndrome. Three groups of seven calves were infected weekly with 10, 50 or 100 Fasciola hepatica metacercariae for 12 consecutive weeks. Maximum glutamate dehydrogenase and gamma glutamyltransferase activities in the plasma were greater with the larger infecting doses but there was no change in plasma clotting times.     

Selenium deficiency

Calvings in a herd infected by Brucella abortus were monitored by cultural examination of milk and mucus and by serological and cultural examination of 5 ml samples of the blood. Out of 129 samples from which culture of B abortus was attempted, only one yielded a positive result.     

Luteinizing hormone secretion in ovariectomized gilts: effects of age, reproductive state and estrogen replacement

Crossbred gilts were ovariectomized (OVX) at 120, 150, 180 and 210 d of age to determine whether various characteristics of luteinizing hormone (LH) concentrations are influenced by age and reproductive state (prepuberal vs postpuberal). All 120-d-old gilts were prepuberal and all 210-d-old animals were postpuberal, whereas gilts 150 and 180 d old included both prepuberal and postpuberal animals. Blood was collected at 15-min intervals for 2 h, 2 d before OVX (d -2), and 2 (d +2), 8 (d +8) and 14 (d +14) d after OVX. Mean LH concentrations for prepuberal gilts were similar among age groups (P greater than .05) on d -2 and +2; however, LH increased (P greater than .05) from d -2 to +2. No change in LH secretion was found in postpuberal gilts during these two periods. After OVX, LH increased from d +2 to +14 in both prepuberal and postpuberal gilts in all age groups. In postpuberal gilts, LH increased linearly (P less than .05) between d +2 and +14; rate of increase accelerated with advancing age (P less than .01). In prepuberal gilts, LH increased in a nonlinear manner, but it did not increase between d +2 and +8. The increase observed in prepuberal and postpuberal gilts after OVX resulted primarily from an increase in magnitude of peak concentrations of LH. Implants of estradiol-17 beta (E2) were used to determine whether the postovariectomy increase in LH is affected differently by E2 in prepuberal and postpuberal gilts during advancing ages.(ABSTRACT TRUNCATED AT 250 WORDS)