Neutrophils are present in milk of cows as a means of suppressing invading pathogens during mastitis. However, the manner by which neutrophils traverse the secretory epithelia is still not clear: do they diapedese between epithelial cells or do they kill epithelial cells to gain entry into milk? We investigated the process of bovine neutrophil diapedesis across bovine mammary gland epithelium in vitro. The bovine mammary epithelial cell line MAC-T, grown on collagen-coated filters, formed a confluent monolayer with characteristic tight junctions, basal-apical polarity, and functional barriers to the dye trypan blue. Neutrophils added on the apical surface of the monolayer were stimulated to diapedese across the epithelium by the addition of Staphylococcus aureus (10(7) colony-forming units/ml) to the basal compartment. Light and transmission electron microscopy revealed the series of events for neutrophil transmigration: accumulation of neutrophils on the surface of epithelial monolayer; projection of pseudopods into intercellular junctions and movement of neutrophils between adjacent epithelial cells; and reapproximation of the lateral epithelial cell membranes and reformation of the apical tight junctions after neutrophils crossed the epithelium. Morphologically, epithelial cell damage caused by neutrophil diapedesis was not evident. This in vitro model provides a two-dimensional epithelial sheet by which neutrophil diapedesis can be qualitatively studied under defined conditions. Results of the study suggest a major mode by which bovine neutrophils diapedese across the alveolar epithelia into milk during mastitis.

Methods available for measurement of plasma lipoprotein-cholesterol concentrations and activities of lipoprotein lipase, hepatic lipase, lecithin:cholesterol acyl transferase (LCAT), and cholesteryl ester transfer protein were adapted for use in cats. A combined ultracentrifugation/precipitation procedure was used to isolate very low-density lipoproteins (VLDL), then to separate low-density lipoproteins (LDL) from high-density lipoproteins (HDL). The reagent used, 92 mM heparin-manganese chloride, provided complete precipitation of LDL with only trace and insignificant contamination by HDL. Efforts to selectively measure lipoprotein lipase activity in plasma, collected after IV injection of heparin, by inhibiting hepatic lipase with sodium dodecyl sulfate were unsuccessful, and the activity of this enzyme was calculated as the difference between total and hepatic lipase activities. The latter was measured in the presence of high salt concentration to inhibit lipoprotein lipase. Cholesterol esterifying activity was identified in feline plasma and was typical of LCAT, in that it was dependent on apolipoprotein A-I as a cofactor. The intra-assay and interassay coefficients of variation for measurement of lipoprotein lipase, hepatic lipase, and LCAT activities were 18.4, 4.6, and 7.2%, and 20.4, 10.7, and 5.3%, respectively. Appreciable cholesteryl ester transfer protein activity was not detected in either undiluted or diluted plasma. These methods were subsequently used to investigate the effects of pregnancy and lactation on lipoprotein metabolism in a group of 10 queens. Plasma concentrations of cholesterol and triglycerides were unaltered during pregnancy, but the concentrations of VLDL-cholesterol increased and those of HDL-cholesterol decreased. During lactation, the concentrations of cholesterol and triglycerides decreased owing to reductions in VLDL-cholesterol and LDL-cholesterol concentrations and continued suppression of HDL-cholesterol. These changes were associated with alterations in the activities of lipoprotein lipase, which increased after parturition, and hepatic lipase, which increased during pregnancy and lactation, that may help explain their metabolic origins. The activity of LCAT remained unchanged.

The bicipital tendons and bursae of 25 healthy adult Quarter Horses were ultrasonographically examined. Cross-sectional images of the right and left bicipital tendons were obtained from each horse, using a 7.5-MHz transducer held in the frontal plane at the point of the shoulder. The bicipital tendon at the point of the shoulder appeared as a bilobate structure overlying the echogenic surface of the humerus. Median distance from the skin surface to the cranial surface of the tendon on the medial sagittal plane of the tendon was 23 mm (range, 16.5 to 30 mm); median distance on the lateral sagittal plane was 14 mm (range, 8.5 to 19 mm). Median distance from the skin surface to the tendon on the midsagittal plane of the tendon was 17 mm (range, 10.5 to 22 mm). Median cranial-to-caudal widths of the lateral and medial lobes of the tendon at their greatest dimensions were 20.5 mm (range, 18 to 27.5 mm) and 16 mm (range, 13 to 20.5 mm), respectively. The median cranial-tocaudal width of the central (midsagittal) portion of the tendon was 10 mm (range, 7 to 13.5 mm). The bicipital bursa was less than or equal to 3 mm wide at all locations at which it was measured. Ultrasonographic imaging was easily performed and allowed evaluation of the bicipital tendon, bursa, and surface of the underlying humerus.

Selenium concentration was measured in paired maternal blood samples and fetal liver specimens collected at a San Joaquin County, Calif, slaughterhouse (beef = 19, dairy = 54) and from bovine aborted fetuses submitted to the California Veterinary Diagnostic Laboratory System (CVDLS; beef = 20, dairy = 20). Of the slaughterhouse samples and specimens, dairy maternal blood selenium concentration was significantly (P < 0.001) higher (mean +/- SD; 0.22 +/- 0.056 micrograms/ml) than that for beef breeds (0.137 +/- 0.082 micrograms/ml). The CVDLS mean maternal blood selenium concentration for the dairy-breed samples (0.192 +/- 0.028 micrograms/ml) was similar to that for the slaughterhouse dairy-breed samples, but was greater than that for the slaughterhouse beef-breed samples. Slaughterhouse mean fetal liver selenium content also was higher (P < 0.001) for the dairy breeds (0.777 +/- 0.408 micrograms/g), compared with the beef breeds (0.443 +/- 0.038 micrograms/g). Mean fetal liver selenium content for slaughterhouse specimens was higher (P < 0.002) than that for the CVDLS specimens (beef, 0.244 +/- 0.149 micrograms/g; dairy, 0.390 +/- 0.165 micrograms/g). At the CVDLS, dairy fetal liver content was greater (P < 0.001) than that for beef breeds. Mean ratio of fetal liver selenium content to maternal blood selenium concentration was 3.53 +/- 1.89 for dairy breeds at the slaughterhouse (liver-to-blood correlation [r] = 0.38), and was 2.11 +/- 1.00 for dairy breeds at the CVDLS (r = 0.31) and 3.43 +/- 1.50 for beef breeds (r = 0.58). Both slaughterhouse breed ratios were significantly (P < 0.002) greater than the CVDLS dairy-breed ratio. On the basis of these results, breed and source location should be taken into account when interpreting selenium values. Fetal liver selenium content should only be used as a screening test and combined with whole blood selenium concentration from clinically normal herdmates to evaluate herd selenium status.

In Zimbabwe, ocular squamous cell carcinoma (OSCC) was frequently observed in 5 breeding herds of Simmental cattle, a Bos taurus breed originating from Switzerland. In these herds, initial signs of OSCC were already noticeable in cattle about 3 years old. Gradually, OSCC prevalence increased, and 36 to 53% of cattle over 7 years old had 1 or more tumors. More tumors developed in Simmental cattle with periorbital white skin than in cattle with periorbital pigmented skin. Other breeds of cattle (eg, Friesian) also are partly white-faced and live in Zimbabwe in a comparable environment; yet, OSCC prevalence was lower in those breeds.

An avirulent live Salmonella choleraesuis culture (SC-54) was evaluated for use as an effective vaccine in preventing salmonellosis caused by S choleraesuis in pigs. Eighty-two pigs, 3 to 4 weeks old, were randomly assigned to 1 of 2 treatment groups, which were designated as either vaccinates or controls. After vaccination, all pigs were examined for fecal shedding of S choleraesuis, rectal temperature, and 10 clinical variables. Significant difference was not detected between vaccinated and nonvaccinated pigs for 14 days (phase I) after intranasal administration of the vaccine. Efficacy and duration of immunity were examined by intranasally challenge exposing respective pigs from either treatment group with a virulent field isolate of S choleraesuis at 2, 8, or 20 weeks after vaccination (phases II-IV). Pigs were again evaluated for 14 days after challenge exposure, and 10 clinical variables and rectal temperature were monitored. Surviving pigs were euthanatized and evaluated for gross lesions, and samples of 7 organs were collected. These organ samples were homogenized, and level of S choleraesuis infection was determined. After virulent challenge exposure during phases II-IV, the clinical status of the SC-54 vaccinates was significantly (P < 0.05) superior to that of nonvaccinates for rectal temperature, feces consistency, behavior, appetite, body condition, and mean score for the 10 clinical variables. Quantitative bacteriologic culture of the tonsil, lung, liver, spleen, mesenteric lymph nodes, ileum, and colon samples indicated consistent reduction of organ colonization in vaccinates; bacteria numbers in the mesenteric lymph nodes, lungs, and ileum were significantly (P < 0.05) reduced. Gross lesions in pigs indicated reduction of pneumonia in vaccinates. Pigs also had consistent weight gain throughout all phases of the study after challenge exposure, although the differences were not significant. In conclusion, a single intranasally administered dose of SC-54 given to 3- to 4-week-old pigs proved to be safe and efficacious and to provide protection to pigs at least 20 weeks after initial vaccination.

Twenty-four, adult, female Beagles were arranged by body weight from greatest to least and allocated to 2 groups of 12 dogs, using random numbers. Dogs were housed collectively in 2 adjacent metal buildings, each divided into 4 rooms measuring 2.1 X3.7 m. Each room was paneled and carpeted and had an access door to the outside with a connecting run that measured 2.1 X 9.1 m. Each run had a surface consisting of 5 cm of pea gravel overlayng 5 cm of sand, and was partially covered by an awning that provided shade at its proximal end. For placement in room/run units, dogs in each of the treated and control groups were allotted to 4 subgroups of 3 dogs each. Each subgroup of dogs was placed in a separate room/run unit. Units containing treatment or control subgroups were alternated to avoid placing identically treated subgroups adjacent to each other. Dogs of subgroups A, C, E, and G were treated with lufenuron monthly at a minimal target dosage of 10 mg/kg of body weight; those of subgroups B, D, F, and H were treated with excipient tablets. Dogs were treated on study days 7, 37, 68, and 98. Each dog was infested with 100 newly emerged, unfed, insectary-reared adult Ctenocephalides felis on each of study days 0 and 2. Thereafter, infestations on all dogs were dependent on continued development of fleas either in the indoor or outdoor environment. Numbers of fleas on each of the treated and control dogs were determined, using a nondestructive counting technique on days 6, 14, 21, 28, 35, 56, 70, 84, 98, 112, and 119. On study day 21 and on each collection day thereafter, numbers of adult fleas recovered from treated dogs were significantly (P < 0.05) fewer than those recovered from control dogs. Proportion reduction of fleas on treated vs control dogs exceeded 90% by study day 35 and 95% by study day 56. Efficacies exceeded 95% on all remaining study days except days 98 (94.4% and 119 (90%). Results of this study indicate that control of flea populations can be achieved in treated dogs approximately 4 to 5 weeks after initial treatment with lufenuron, and that continued monthly treatments will maintain effective control of flea infestations. Adverse reactions or side effects to treatment with lufenuron were not observed in dogs after treatment at any time throughout the study.

A surgical technique was developed for implanting a flexible polyurethane cannula in a lateral ventricle in the brain of calves. Initially, measurements were made on 25 calves at necropsy to develop equations for calculating coordinates for cannula placement. The distance (cm) caudal, in the sagittal plane, from the coronal suture line to the center of a hole to be drilled in the parietal bone of the skull was: 0.73 + (0.00925 X body weight [kg]). The distance (cm) lateral from the midline to the center of the hole to be drilled was: 0.018 + (0.6464 X distance caudal). The depth (cm) from the surface of the skull to the dorsal surface of the lateral ventricle was: 2.29 + (0.0159 X body weight [kg]). Surgery was subsequently performed on 17 calves. A 5-mm-diameter hole was drilled through the skull with a hand trephine at coordinates derived from the aforementioned regression equations. A polyurethane cannula (total length, 30 cm; 1 mm ID; 2 mm OD) covering a stainless-steel 20-gauge blunt-tipped needle (stylet) was lowered through the brain and into a lateral ventricle at an angle of 20.5 degrees relative to the frontal bones of the skull. The blunt-tipped needle was then removed, and CSF was allowed to drip from the cannula to verify placement. One stainless-steel screw was inserted 0.6 cm medial, and another was inserted 0.6 cm caudal to the hole in the skull. The area around the cannula, bone screws, and hole in the skull was covered with dental acrylic (approx 2 cm in diameter) to stabilize the cannula. With minimal restraint of calves, injection of substances into and withdrawal of CSF from a lateral ventricle of the brain were possible in most calves for at least 6 weeks after surgery was performed.

A total of 22 clinical streptococcal isolates, predominantly Streptococcus zooepidemicus, associated with endometritis in horses were tested for their ability to withstand the natural bactericidal properties of freshly obtained blood. During a 3-hour incubation in blood from a single horse, 8 of these isolates survived and grew; the remainder were killed. To determine whether this ability to grow extended to blood of other horses, 5 of these growing isolates were tested for their ability to grow in the blood of 5 additional horses. The same 5 horses were used for each isolate. The isolates grew in blood of some of the horses, but were killed in blood of the others. However, the horse's blood that mediated killing was different for each isolate. Killing required leukocytes, but the specificity for killing appeared to reside in plasma, although plasma by itself was not bactericidal. Heat-stable and heat-labile components in plasma, interpreted as antibody and complement, respectively, appeared necessary for killing. Isolates that could grow in fresh blood lost this ability after 10 passages in artificial media. Results of these experiments of phagocytosis in fresh blood may provide helpful insights into the phagocytosis of S zooepidemicus in equine uterine f1uid.

Milk antimicrobial residues are a serious concern for the dairy industry. Residues of the tetracycline family of antimicrobials have been reported in market milk by investigators, using radioimmunoassay and microbial receptor technology (hereafter referred to as the Charm II test). In response to these reports, an investigation was conducted to determine the potential of 3 extra-label routes of oxytetracycline (OTC) administration to cause milk residues above the Food and Drug Administration safe value of 30 parts per billion (ppb). Lactating Holstein cows were administered OTC once by use of 1 of 3 routes: IV at 16.5 mg/kg of body weight (n = 6); IM at 11 mg/kg (n = 6); and intrauterine (IU) at 2 g in 500 ml of saline solution/cow (n = 6). Duplicate milk samples were collected at the milking prior to drug administration and for the next 13 milkings at 12-hour intervals. Concentrations of OTC in milk samples were analyzed by use of the Charm II test for tetracyclines (limit of OTC detection, approx 5 ppb) and were compared with concentrations determined by use of a high-performance liquid chromatography (HPLC) method (lower limit of OTC quantitation, approx 2 ppb). The potential for milk OTC residues above the Food and Drug Administration safe value of 30 ppb after treatment was considerably greater for the IV and IM routes, compared with the IU route. Mean peak OTC concentrations in milk at the first milking after treatment for the HPLC and Charm II tests were approximately 3,700 to 4,200 ppb for the IV route, 2,200 to 2,600 ppb for the IM route, and 186 to 192 ppb for the IU route, respectively. Pharmacokinetic analysis, based on milk OTC concentrations, indicated that the area under the curve (AUC) and milk maximal concentration (Cmax) differed significantly (P < 0.001) among routes of administration. The AUC was similar for IV and IM administrations; values for both were greater than the AUC for IU administration. The Cmax was greatest for IV, intermediate for IM, and least for IU administration. There were significant (P less than or equal to 0.01) differences in AUC between assay methods (Charm II vs HPLC) for the IV route. Concentrations of OTC in milk determined by the Charm II test were often greater than those determined by HPLC. Administration of OTC to lactating cows via these routes is extra-label drug use. Failure to withhold the product from early milkings of cows administered OTC by the IV or IM route should be considered a potential cause of OTC residues in market milk. Milk from nearly all cows contained OTC (< 30 ppb), the Food and Drug Administration safe level, by 120 hours after OTC administration. Use of appropriate withholding times and antibiotic residue testing is indicated to avoid OTC residues.