Seven mature dairy cows from 6 herds were obtained with history, clinical signs of disease, and laboratory findings suggestive of advanced paratuberculosis. A surgically implanted collection chamber was used to obtain peripheral tissue fluid. Blood, mammary gland flush fluid, and collection chamber flush fluid (CCFF) samples were obtained 6 times over a 2-week period from each cow. Mononuclear cell-rich portions of these fluids obtained by gradient centrifugation were submitted for bacteriologic culture of Mycobacterium paratuberculosis and for total and differential cell counts. Bacteriologic culture of feces for M paratuberculosis and complete necropsy performed on each cow at the conclusion of the study confirmed the diagnosis of paratuberculosis. Numbers of tissue macrophages obtained from CCFF samples were lower than expected. Mean (+/- SD) differential count of tissue macrophages collected from CCFF was 65.57 (+/- 23.39). Mean calculated tissue macrophages (total cell count X differential count) collected from CCFF samples was 623.1 (+/- 784.55) cells/microliter. Mycobacterium paratuberculosis was isolated from 1 of 42 (2.4%) collections of mononuclear cell-rich portions of plasma and from 2 of 42 (4.8%) CCFF samples. Mycobacterium paratuberculosis was not isolated from any collections of mammary gland flush fluid. The collection and processing techniques used in this study did not enhance detection of M paratuberculosis infection in cows with advanced paratuberculosis, beyond that of ileocecal lymph node biopsy or fecal culture.

Enzyme-linked immunosorbent assay and flow cytometric methods to screen hybridoma culture supernatants for antibodies to bovine neutrophils (surface antigen-specific) were optimized. Sensitivity of the 2 methods was compared. A panel of 14 murine monoclonal antibodies (MAB) to surface antigens of bovine polymorphonuclear neutrophilic leukocytes (neutrophils) was produced by hybridoma technology, and their isotypes were determined by whole-cell ELISA. Monoclonal antibody reactivity with neutrophils, eosinophils, and lymphocytes isolated on phosphate-buffered saline solution and on Ficoll-sodium diatrizoate were compared. Biochemical characterization of antigens recognized by MAB was performed by immunoblot analysis. Neutrophil plasma membranes were isolated on sucrose gradients (20, 32, and 50%) and purified for polypeptide characterization. Neutrophil surface proteins were characterized by external labeling with 125I. The flow cytometric method was proven to be more sensitive and rapid than ELISA to screen hybridoma supernatants. This method allowed light-scatter gating of live neutrophil populations for analysis, which eliminated nonspecific binding of antibodies to contaminating cells and dead neutrophils. The optimal conditions for flow cytometric analyses were 5 X 10(5) neutrophils and 1 microgram of fluorescein-labeled F(ab')2/assay as the second antibody. The optimal conditions for hybridoma screening by ELISA were neutrophil concentration of 2.5 X 10(5) well, using a 96-well polystyrene microtitration plate as solid support, and 2,2'-azino-di[3-ethyl-benzthiazoline sulfonate (6)] with H2O2 as the chromogenic substrate. Tissue culture plates as solid support and 3,3', 5,5'-tetramethyl benzidine, with H2O2 as the chromogenic substrate, were equally as sensitive. Panel MAB reacted differently with neutrophils, eosinophils, and lymphocytes. Isolation of these cells from blood on Ficoll-sodium diatrizoate generally did not alter MAB reactivity. Coomassie blue-stained gels of neutrophil plasma membrane proteins contained about 25 polypeptide bands, 13 of which were major bands. Autoradiography revealed about 11 surface proteins, 5 of which were heavily labeled with 125I. Monoclonal antibody S7G8 identified a 65-kd protein and MAB S8G10 identified 65- and 70-kd proteins. On the basis of molecular weight, MAB S7G8 and S8G10 are comparable to human CD15, CD16, and CD64 molecules. The MAB generated in this study are potential candidates to discern bovine neutrophil function and heterogeneity.

Single doses (2.2 mg/kg of body weight) of phenylbutazone (PBZ) were administered IV to 6 neonatal horses (5 to 17 hours old at time of dosing). Plasma concentrations of PBZ and its metabolite oxyphenbutazone were monitored serially for 120 hours after drug administration. Pharmacokinetic variables were calculated, using 1- and 2-compartment open models. Descriptive equations from the best model for each foal were then used to derive model-independent variables describing PBZ disposition. Median volume of distribution at steady-state was 0.274 L/kg (range, 0.190 to 0.401 L/kg). Median terminal half-life was 7.4 (6.4 to 22.1) hours, and median total plasma clearance of PBZ for foals in this study was 0.018 L/kg/h (range, 0.013 to 0.038 L/kg/h). Volume of distribution was larger, half-life was longer, and total clearance was lower, compared with similar values reported for administration of PBZ to adult horses.

Tissue specimens and serum samples obtained from adult budgerigars in various stages of reproduction housed in an aviary with enzootic avian polyomavirus (APV) disease were examined by means of polymerase chain reaction techniques for APV DNA. Although the birds were apparently healthy, APV DNA could be detected in all 40 birds examined (inapparent infection rate, 100%). Viral DNA was found in most organ systems examined. Analysis of data suggested that organ virus concentrations were lower in breeding than in nonbreeding birds. Serum samples from 144 birds were examined for virus-neutralizing (VN) antibody. All serum samples had detectable VN antibody titers. Determining VN titer had a sensitivity of 100% for detection of APV infection in birds and was more sensitive than analysis of droppings by use of polymerase chain reaction techniques to detect APV infection in 6-month-old birds. Analysis of the data suggested that lower VN antibody titers were associated with longer duration of continuous breeding.

Selective parathyroidectomy (PTX) is preferred to thyroparathyroidectomy (TPTX) when specific effects of parathyroid hormone depletion are being studied. However, because of the anatomic proximity of thyroid and parathyroid glands, TPTX often is performed, leaving animals depleted of thyroxine (T4) and calcitonin as well as parathyroid hormone (PTH). In the present study, six normal dogs had parathyroid tissue and about seven-eighths of thyroid tissue removed. This quantity of thyroid tissue was inadequate to maintain normal serum T4 concentrations, despite allowance of 168 days for thyroid recovery. Five of six dogs with reduced renal mass had successful selective PTX and normal serum T4 concentrations at 28 days, when one-half or more of thyroid tissue was spared. We conclude that with attention to the surgical technique, selective PTX can be achieved in a high percentage of dogs and sufficient thyroid tissue spared to maintain euthyroidism.

In vitro transferability of penicillin, streptomycin, tetracycline, and erythromycin resistance from coagulase-negative staphylococci to Staphylococcus aureus and among the former species of bovine mammary gland origin was examined by bacterial mating on filters and by mixed-culture matings in broth and in skim milk. One hundred twenty-six (42 each on filter, in broth, and in skim milk) matings were performed among 37 isolates of different Staphylococcus species. Transfer of resistance to penicillin, tetracycline, or erythromycin was not detected. Of 51 matings performed to determine streptomycin-resistance transfer, 9 (3 each on filters, in broth, and skim milk) were successful. Nine strains representing 3 species of coagulase-negative staphylococci were tested as prospective donors of streptomycin resistance. Of these, 2 strains could transfer streptomycin resistance. A double-resistant donor, S hominis, not only transferred its streptomycin resistance to an S chromogenes strain lacking resistance, but also to an S aureus strain already carrying penicillin and tetracycline resistance. The transfer of streptomycin resistance from the donor S hominis, harboring 2 plasmids, to a plasmidless S chromogenes recipient strain was associated with apparent acquisition of the smaller plasmid of the donor by the recipient. The single-resistant donor, S epidermidis 681A, transferred streptomycin resistance to a tetracycline-resistant S aureus recipient. This strain however, failed to transfer its streptomycin resistance to another S aureus, 2 S hyicus, and 1 S xylosus recipient. Frequency of transfer of streptomycin resistance ranged from 1.1 X 10(-5) to 1 X 10(-4). When transfer of resistance was successful, attempts were made to characterize the transfer process. Conjugation appeared to be the mode of streptomycin-resistance transfer. Transfer of resistance between staphylococci of bovine mammary gland origin appears to be fairly uncommon. However, in view of the limitations of the procedures used, additional in vitro and in vivo work is needed to further assess the role of coagulase-negative staphylococci in dissemination of antibiotic resistance.

Streak retinoscopy was performed by 5 ophthalmologists on 256 eyes (191 dogs) to determine their postoperative refractive state after cataract extraction. Aphakic and pseudophakic eyes that had been implanted with 1 of 5 intraocular lenses (IOL) with dioptric powers ranging from +14.5 to +38 diopters (D) were studied. By use of ANOVA, breed and body type of dog and individual performing refraction were found to have no detectable effect on final refractive state. Mean refractive state of aphakic eyes was +14.4 +/- 2.10 D. Mean refractive state for different IOL powers was as follows: +14.5 D IOL = +11.54 +/- 1.18 D (n = 13); +30 D IOL = + 5.15 +/- 1.18 D (n = 105); +34.0 D IOL = +3.5 D (n = 1); +36 D IOL +2.34 +/- 0.73 D 9 (n = 61); and +38 D IOL = +1.41 +/- 0.56 D (n = 28). Residual hyperopia ranged from +0.5 D to +2.5 D with +38 D IOL, and no eyes were myopic (overcorrected) by use of any of the IOL studied. linear regression analysis of refractive state on IOL power for aU dogs predicted that dioptric strength of +41.53 D was necessary to best approximate emmetropia for the population as a whole. Body type of the dog had only slight effect (< 1.0 D) on predicted optimal IOL power. Further linear regression analysis of the 7 breeds studied predicted variations from +39.62 to +43.14 D in IOL powers necessary to approximate emmetropia. Results of the study support the routine use of canine IOL with dioptric strength of approximately +41.5 D in circumstances in which preoperative biometry and keratometry are not practical. The findings further suggest that, for the specific population of dogs studied, most of the dogs could be corrected to near emmetropia by use of a small range of IOL dioptric strengths, irrespective of body type or breed.

After a detailed anatomic study to determine puncture sites, 10 cadaver elbows from 5 dogs were examined arthroscopicalty to study the normal intraarticular anatomy, as viewed from the medial side. Subsequent dissection revealed absence of neurovascular injury and only minor iatrogenic damage to the cartilage. The technique was clinically applied and evaluated in 13 dogs (26 joints). The dogs recovered without complications. The technique proved to be safe and reliable for direct examination of nearly the entire joint. More specifically, it allowed systematic inspection of the medial and lateral humeral condyles, the medial and lateral coronoid processes, the caudal and middle parts of the head of the radius, the olecranon (including the anconeal process), and the medial collateral ligament.

To investigate the feasibility of using changes in body or mammary temperature to detect mastitis, radiotransmitters were implanted midway between rear udder quarters and in the peritoneal cavity of 5 Holstein cows (1 to 3 months in lactation) housed in an environmental chamber (16 +/- 2 C; lights on 7:00 AM to 11:00 PM). After a 6-week control period, Escherichia coli endotoxin (0.5 mg) was injected after the morning milking into left rear teat cisterns via the teat canal. Wisconsin mastitis test score and somatic cell count in all quarters increased significantly (P < 0.01) by the next milking. Effects were greatest in the endotoxin-exposed quarters. Milk yields for all quarters decreased significantly (P < 0.01) by the first milking after endotoxin injection. Udder and body temperatures at milkings were similar and were not affected by treatment. When temperatures were averaged for the 5 cows for each of 120 time points/d, average temperatures, relative to time of injection of endotoxin, were increased by 0.5 C above baseline at 2.75 hours, peaked at + 2.9 C at 6.50 hours, and remained high through 9.25 hours after injection. Power spectra calculated for individual cows on a daily basis universally indicated an increase in power at low frequencies on the day of injection. Subsequently, Streptococcus agalactiae (200 colony-forming units) was injected into right rear teat cisterns. Wisconsin mastitis test score increased at the second milking after injection. Cell count and quarter milk yield decreased by the third milking. As with endotoxin, injection of S agalactiae could not be detected via a change in temperature at milkings. Of the 5 cows, 3 had a peak in temperature after injection of S agalactiae. Average temperatures for these 3 cows relative to time of injection, were increased by 0.5 C above baseline at 24.25 hours, peaked at + 1.4 C at 26.25 hours, and remained high through 28.75 hours after injection. Power spectra calculated for the day in which a temperature peak was detected for these 3 cows indicated an increase in power at low frequencies, compared with spectra for all other days. Similar increases in power were also detected for the 2 cows that did not have temperature peaks. When clinical signs of mastitis are obvious at milking, there is little advantage of using body temperature for detection of infection. When clinical signs are not obvious, body temperature is often only minimally increased. Thus, monitoring body temperature at milkings adds little to the ability to detect mastitis. Of more interest is the ability to detect transient temperature increases that often develop in association with less-severe infections. Also, as early treatment increases the likelihood of successful treatment, detection of the onset of temperature increases would be advantageous for treatment of severe infections. Detection of a transient temperature peak requires taking temperature readings every 2 hours. To detect mastitis when a temperature peak does not occur requires measurement every 15 minutes to calculate power spectra. The ability to detect the onset of acute clinical infections and subclinical infections, using frequent temperature readings, indicates that development of a practical radiotelemetry system for use on farms may be warranted, depending on cost. The added potential of using body temperature to monitor general health and to detect estrus enhances the economic feasibility of developing such a system.

Fourteen cats were inoculated orally with 1 of 2 infective doses of Toxocara canis to induce eosinophilia. Cats were subsequently challenge exposed twice via intraperitoneal injection with 1 of 2 T canis antigen preparations. Peritoneal lavage was performed 2 days after antigenic challenge exposure, and eosinophils in the peritoneal lavage fluid were quantified. None of the cats developed clinical signs of disease after infection. All cats developed peripheral eosinophilia after infection. Significant (P < 0.05) difference in mean eosinophil count from the lavage fluid was observed between lavage 1 (prechallenge exposure) and lavages 2 and 3 (postchallenge exposure) in both groups of cats. Significant difference in eosinophil count was not found between cats given different doses of eggs. After initial challenge exposure, significantly (P < 0.05) more eosinophils were obtained from cats given antigen preparation 2 (prep-2) than from those given antigen prep-1. This difference was no longer observed after the second challenge exposure with higher doses of either antigen prep-1 or prep-2. In cats given antigen prep-2, significant difference was not found between lavages 2 and 3. However, in cats given antigen prep-1, eosinophil count was significantly (P = 0.005) greater in fluid obtained from lavage 3, compared with eosinophil count from lavage 2. Mean +/- SEM percentage of eosinophils in the fluid from lavage 3 in all cats was 70.8 +/- 2.2%. Other cell types included macrophages, neutrophils, lymphocytes, and mast cells. Gross postmortem findings were mild. One- to 3-mm nodular white foci of inflammation were observed on the serosal surfaces of the liver, spleen, kidneys, and omentum. Microscopic examination of tissues revealed pulmonary artery hypertrophy (n = 4), eosinophilic peribronchitis and perivasculitis (n = 10), mild granulomatous interstitial nephritis (n = 6), interstitial pancreatitis (n = 1), focal lymphocytic myocarditis (n = 1), focal eosinophilic granulomatous hepatitis (n = 1), and eosinophilic hyperplasia of bone marrow (n = 14). Large numbers of eosinophils could be harvested from the peritoneal cavity of cats inoculated orally with 500 embryonated T canis eggs and subsequently challenge-exposed intraperitoneally with preparations of parasite antigens. After the second challenge exposure, at least 108 eosinophils could be harvested from each cat, yielding eosinophils in the quantity required to begin isolation of granule constituents.