Sixteen Holstein cows were used to test the effect of postmilking teat treatment on colonization and intramammary infection by Staphylococcus aureus on chapped teats. Treatments were (1) chapping the teat and using 1% I2/10% glycerin postdip solution, (2) 1% I2/10% glycerin postdip solution on nonchapped teats, (3) chapping the teat and using 10% glycerin postdip solution, (4) chapping the teat and not using a postdip solution. All mammary glands were free of S aureus teat skin colonization and intramammary infection at the start of the study. Teats selected for chapping were dipped in 1N NaOH prior to 3 applications of S aureus broth culture; cultures were applied at 12-hour intervals on all teats. Treatments were applied after each milking for 30 days and were initiated after the second broth dip. Teat skin swab specimens and milk samples were collected before treatment application. Teat skin condition was scored daily. Nonchapped teats (treatment 2) did not support skin or orifice colonization by S aureus. Treatment-1 teats healed most rapidly and supported less colonization in skin and orifice than did treatment-3 and -4 teats. Teat skin scores and skin colonization were lower for treatment-3 than treatment-4 teats. A correlation between teat skin colonization and teat skin conditions was found. Two intramammary infections were found in treatment-4 quarters and 1 in a treatment-3 quarter. On the basis of our findings, we concluded that poor teat skin condition will more readily support S aureus colonization, that a dip of 1% I2 with glycerin helped reduce S aureus colonization and was associated with faster healing, and that glycerin in teat dips may be of value in preventing colonization by S aureus and in promoting healing.

The purpose of the study was to determine whether there were circadian variations in serum osteocalcin in normal horses and to determine whether it was important to regulate the time of blood sampling in clinical investigations. Osteocalcin or bone Gla-protein (BGP), alkaline phosphatase, total calcium, phosphate and total protein were studied over a 24 h period. Blood samples were taken every 60 min from nine adult Standardbred horses. There was a correlation between serum levels of alkaline phosphatase (r = 0.3, p < 0.01), phosphate (r = 0.42, p < 0.01) and serum osteocalcin levels. There was a very marked individual effect on serum levels of osteocalcin and alkaline phosphatase (p < 0.01). This effect was present for phosphate levels but not significant for total calcium. The individual effect was lower and time effect was higher for serum osteocalcin if the subjects were divided into two age groups, one of horses of five years or less (n = 4) and a second group older than five years (n = 5). In both groups a circadian rhythmicity was observed. Serum osteocalcin showed a biphasic pattern. Levels were constant during daytime (light period) and underwent significant variations during the night (dark period), going through a nadir at 2000 h and through a maximum peak at 0500 h. It was concluded that in normal horses the blood osteocalcin level follows a circadian variation. Also daytime (light period) seems to be the more appropriate period for blood sampling.

To assess the suitability of sheep for exercise studies, the effect of incremental exercise and conditioning on oxygen consumption was studied. Six sheep were adapted to a treadmill and subsequently trained 8 weeks. The sheep were then studied, in random order, using 3 incremental exercise protocols (EX-1, EX-2, and EX-3). The protocols were chosen to approximate high (EX-1), moderate (EX-2), and low (EX-3) intensity exercise by varying treadmill speed and incline. The sheep were then conditioned for an additional 12 weeks and retested on the EX-2 protocol. During exercise, oxygen consumption, gas exchange ratio (R), and rectal temperatures (Tb) were recorded. All 3 protocols resulted in significant increases in oxygen consumption, R, and Tb (P < 0.05). Maximum oxygen consumption for EX-1, 49.9 +/- 5.0 ml/min/kg of body weight, was significantly greater than maximum oxygen consumption for EX-2 and EX-3, 37.8 +/- 6.5 and 42.3 +/- 6.0 ml/min/kg, respectively (P < 0.05), whereas maximum R and maximum Tb were similar. After the additional 12-week conditioning, time on the treadmill increased 40% from 9.58 +/- 0.87 to 13.4 +/- 0.44 minutes, and maximum oxygen consumption increased 27% to 48.1 +/- 9.1 ml/min/kg. These data indicated that maximum oxygen consumption varied with intensity of the exercise, 12 weeks of maximal exercise conditioning was sufficient to produce a measurable training effect (ie, increase endurance and maximum oxygen consumption) and sheep are suitable for maximal exercise studies where oxygen consumption measurements are desired.

The effect of various confinement conditions on physical fitness in dogs was evaluated. Eighteen 9.5- to 10-month-old female purpose-bred Beagles were maintained individually for 3 months at a time in 1 of 6 confinement conditions: Condition A--an outdoor housing area with a conventional dog house and free access to a 6.1 X 9.1-m pen; condition B--outdoor kennel with a conventional dog house and free access to a 1.8 X 6.1-m run; condition C--indoor environmentally controlled 1.2 X 3.66-m run; condition D--0.9 X 1.2 X 0.84-m conventional laboratory cage in an indoor environmentally controlled room; condition E--0.9 X 1.2 X 0.84-m conventional laboratory cage in an indoor environmentally controlled room with treadmill exercise (7 km/h at a 10% grade) for 30 min/d, 5 d/wk; condition F--0.71 X 0.86 X 0.69-m conventional laboratory cage in an indoor environmentally controlled room. During the final week of each 3-month interval, muscle succinate dehydrogenase enzyme activities and submaximal exercise heart rates (during treadmill exercise) were determined to estimate physical fitness. Also, 5 days after being moved into a different housing condition, blood samples were collected for plasma cortisol determination. The type of confinement condition for dogs had little effect on muscle succinate dehydrogenase activity, but had a modest effect on submaximal exercise heart rates of dogs. At the fifth and tenth minutes of the treadmill exercise period, heart rates of dogs maintained in the smallest cages (condition F) were higher than those of dogs maintained in outside pens and runs (conditions A and B), indicating decreased fitness in the dogs maintained in the smallest cages. Differences in heart rates were not detected among dogs in other conditions. The confinement conditions used in this study had no detectable effect on plasma cortisol concentrations. We concluded that neither cage or pen size nor a regular mandatory exercise program substantially impacted on physical fitness of laboratory confined dogs, as long as the cages complied with federal standards and guidelines. Dogs maintained in substandard cages did have modest decreases in fitness.

To compare the effects of milk stasis and milk flow on Brucella abortus infection of the mammary gland under the same systemic conditions, primiparous goats (n = 5) were inoculated IV with B abortus on the day of parturition, and suckling by their neonates was restricted to one mammary gland. Goats were euthanatized and necropsied at 3 weeks after inoculation, and milk, mammary glands, and supramammary lymph nodes were evaluated by bacteriologic, histologic, and immunoenzymatic staining techniques. Nonnursed mammary glands had high titers of brucellae in milk, moderate interstitial mastitis, and brucellar antigen in macrophages located primarily in alveolar and ductal lumina. Brucellae often filled the macrophage cytoplasm. In contrast, nursed mammary glands had fewer brucellae in milk, minimal inflammatory changes, and no detectable brucellar antigen in histologic sections. Hyperplastic changes were only seen in supramammary lymph nodes draining nonnursed mammary glands; these contained more brucellae than lymph nodes draining nursed mammary glands. These studies show that milk stasis may be the sole cause of increased susceptibility of nonnursed mammary glands to B abortus infection.

A flow cytometric procedure was evaluated to measure the oxidative burst activity (hydrogen peroxide formation) of bovine neutrophils. The method involves measuring the oxidation of intracellular dichlorofluorescin to fluorescent dichlorofluorescein (DCF). Phorbol myristate acetate (PMA) was used to perturb the neutrophil plasma membrane. The sources of variation introduced into the DCF assay were also examined. The sources of variation were attributable to the isolation of neutrophils from blood, variation between duplicate assays and duplicate flow cytometric determinations of oxidative product formation, variation in neutrophil oxidative product formation among cows, and the variation (over repeated daily and weekly neutrophil isolations) in neutrophil oxidative product formation. A final objective was to determine effects of dexamethasone on oxidative product formation, and whether differences existed between blood and mammary neutrophils in oxidative product formation. There was an increasing trend in the formation of DCF with increasing time of incubation and with increasing PMA concentration. Increasing the concentration of PMA decreased lag time and increased the rate of oxidative product formation. The increase in DCF formation was statistically significant up to a PMA concentration of 10 ng/ml. This concentration was considered optimal for bovine neutrophils. Examination of the sources of variation indicated that (i) the neutrophil isolation technique was a major source of variation (17.2 to 28.4% of the total variation), and that more than one neutrophil isolation within a cow would be required to obtain an accurate estimation of DCF formation in neutrophils; (ii) duplicate assays and duplicate readings on the flow cytometer accounted for < 0.05% of the total variation and would not be necessary when performing the DCF assay; (iii) large variation (62.4 to 70.8%) existed among cows in neutrophil oxidative product formation, indicating that any treatment being compared should be done either within or preferably repeated across a large number of cows; and (iv) the variation over repeated daily (0.3%), but not weekly (19.6%) determinations of neutrophil oxidative product formation, were small enough to allow for the evaluation of major physiologic and environmental effects. Intramuscular administration of dexamethasone (50 microgram/ kg of body weight) resulted in an approximate 80% decrease in neutrophil oxidative product formation. Oxidative product formation was 75% less for neutrophils isolated from mammary secretions when compared with neutrophils from blood. These results indicated that the DCF procedure was responsive to factors known to interfere with oxidative metabolism of bovine neutrophils.

The third carpal bone (C3) was collected from both forelimbs of 27 Thoroughbreds. On the basis of age, training, and history, specimens were assigned to 1 of 5 groups: yearling, untrained horses (group 1, n = 4); 2- to 3-year-old, untrained horses (group 2, n = 7); trained 2-year-old horses (group 3, n = 6); trained 3-year-old horses (group 4, n = 6); and 3-year-old, trained horses with carpal pathologic features (group 5, n = 4). A transverse section of subchondral bone 5-mm thick was cut in a precise fashion 10 mm below the proximal articular surface of all specimens. After high-detail radiography was done, indentation testing was performed on the proximal surface of the section at points 5 mm apart. The stiffness of the subchondral cancellous bone was determined from the slope of the load vs displacement curve. Topographic plots of stiffness measurements were compared with radiographs of each specimen. Point determinations were averaged to derive measures for the radial and intermediate facets, and for regions 5, 10, 15, and 20 mm from the dorsal margin of C3. Area fraction (1-p; p = porosity) was measured for the radial and intermediate facets, using an automated image analysis system. Significant (P < 0.05) increases in stiffness and area fraction were found in the C3 from trained horses (groups 3 to 5), compared with untrained horses (groups 1 to 2). Stiffness and area fraction of the radial facet of pathologic C3 were significantly higher than the same variables measured in C. from any other group. A typical profile of regional subchondral stiffness was identified in C3 from normal horses, with maximal stiffness measured 10 mm from the dorsal articular margin. A different pattern was found in pathologic C3, with significantly greater stiffness 15 and 20 mm from the dorsal articular margin when compared with normal horses. A highly significant (P < 0.0001) direct linear correlation between stiffness and area fraction at the radial facet was found. Topographic and radiographic analysis demonstrated good correlation between stiffness and radiographic density of the bone sections. The observed patterns of normal and pathologic C3 were contrasted. In particular, a large gradient in sub-chondral stiffness was identified in pathologic C3 at the dorsomedial aspect of the bone.

Kittens are the principal disseminators of Toxoplasma gondii. They can shed > 10(8) oocysts in the feces after initial infection with bradyzoites in tissue cysts. Thereafter, most kittens develop protective immunity and do not shed oocysts again if they are reinfected. Bradyzoites of a T gondii mutant, designated T-263, were used to vaccinate kittens. Their use did not result in oocyst shedding, but successfully prevented 84% (31/37) of the kittens from shedding oocysts when challenge exposed with a normal isolate of T gondii. Vaccination of outdoor-roaming cats and kittens would be a useful public health measure to prevent transmission of toxoplasmosis near homes, on farms, and in zoos. It is anticipated that several years will be required for a lyophilized bradyzoite vaccine to be ready for licensing and possible commercial availability.

Forty 3- to 17-week old domestic ferrets, including 2 gnotobiotes, were inoculated orally and/or rectally with 10(6), to 10(9) colony-forming units of 1 or more of 4 strains of Campylobacter jejuni, 3 of mink and 1 of human origin. Feeding or gavage of any of the 4 strains, in milk or broth, with or without preinoculation sodium bicarbonate treatment to neutralize stomach acid, induced colonization in 38/40 ferrets; diarrhea lasted 2 to 4 days in conventional kits, 6 days in gnotobiotes. Bacteremia was detected in 4 of 18 tested, 2 to 5 days after inoculation. Two strains caused no more severe disease or prolonged colonization after 3 serial IV passages in kits than they did before passage. Multiple inoculations with a given strain resulted in progressively briefer colonization and milder disease, but subsequent inoculation with a different strain induced colonization and gastrointestinal disease similar to a primary infection. Five kits inoculated rectally after 4 previous homologous inoculations were resistant to colonization as well as to disease. Agglutinin titers of ferrets inoculated orally or rectally once were low or undetectable, but increased in response to repeated inoculation. Pretreatment with a 1% formalin enema caused mild colon irritation without clinical or histologic evidence of proliferative colitis in ferrets concurrently inoculated orally and/or rectally, whether or not they had preexisting antibodies to any strain of C jejuni. Histologic examination of tissues revealed leukocytic infiltration of intestinal lamina propria in 29 of 35 infected kits and 5 of 8 noninfected controls, and cryptosporidiosis in 5 infected kits plus 1 control. Examination of silver-stained sections of intestine from 15 infected ferrets revealed Campylobacter-like organisms on the surface of, but never inside, epithelial cells. The lack of characteristic gross or histologic lesions suggested that C jejuni is not, by itself, responsible for proliferative colitis in ferrets.

Inclusion of lactose in the diets of chickens has been determined to reduce cecal colonization with Salmonella typhimurium. We hypothesized, therefore, that dietary lactose may be a practical means for reducing the prevalence of Salmonella contamination of chicken products. Because some strains of Salmonella are atypical and ferment lactose, we investigated the effects of dietary lactose on cecal colonization with lactose-fermenting S typhimurium. Broiler chicks were inoculated intracloacally with Lac+ S typhimurium selected for resistance to novobiocin and rifampicin. The chicks also were inoculated orally with certain anaerobes that do not effectively inhibit colonization by S typhimurium, but do appear essential for lactose mediated inhibition of cecal colonization. Control chicks were not given dietary lactose, and chicks in the experimental group were fed a diet containing 7% lactose. Enumeration of Lac+ S typhimurium in cecal contents revealed dietary lactose to be effective at controlling this organism. Control was correlated with changes in cecal pH and increases in undissociated volatile fatty acids, especially propionic acid.