Actinobacillus (Haemophilus) pleuropneumoniae plasmids were characterized and classified. They were isolated from A pleuropneumoniae strains different in serotype, year isolated, or location from which isolated. Six of 8 plasmids encoded streptomycin (Sm) and sulfonamide (Su) resistance (SmSu). One of the other plasmids, pVM105, encoded ampicillin (Ap) resistance and another, pHMO, encoded no drug resistance. All SmSu plasmids were transferred to Escherichia coli strains by transformation. Among them, pABO and pMS260 were 8.1 kb and incompatible with each other; they were stable in E coli. The other SmSu plasmids, pHM1, pVM104, pVM106, and pKD25, were 4.3 kb and did not replicate stably in E coli. The former SmSu plasmids were mobilized in E coli strains by a plasmid RP4, which belonged to incompatibility (Inc) group P, but the latter plasmids were not. Further, each 8.1-kb SmSu plasmid and each 4.3-kb plasmid had the same respective restriction pattern. These results indicated that there were at least 2 types of SmSu plasmids in A pleuropneumoniae. The 2 types were classified in 2 groups: Hl(pMS260 and pABO) and H2(pHM1, pVM104, pVM106, and pKD25). The Hl and H2 plasmids belonged to a different Inc groups, and H2 plasmids belonged to a different Inc group from that of pHMO and pVM105.

On the basis of serologic test results and isolation of leptospires from mature cattle, distribution and prevalence of Leptospira interrogans serovars and genotypes were compared by state and region of the United States. Relationships between isolation rate and month of sample collection, mean regional temperature, and mean regional precipitation were examined. Isolation rate and seroprevalence were significantly (P < 0.001) higher for southeastern, south central, and Pacific coastal regions than for other regions of the United States. Isolates of genotypes hardjo-bovis A and kennewicki A and B, and of serovar grippotyphosa appeared to be randomly distributed. Genotype hardjo-bovis B isolates came from a southern area of the country that extends from Georgia to New Mexico. To the authors' knowledge, this is the first recorded isolation of serovar hardjo from Hawaii. Although significant relationship was not documented between isolation rate and month or season of the year, seroprevalence for summer, fall, and winter was significantly (P < 0.001) higher than that for spring. Regional isolation rate was related more to mean temperature (r = 0.83; P < 0.05) than to mean precipitation amount (r = 0.34; P > 0.50).

Chemotaxis under agarose was evaluated to establish an assay system and to characterize chemotacticresponses of canine neutrophils. A method for the measurement of canine neutrophil chemotaxis was established, with optimal responses obtained with agarose containing 10% pooled canine serum, a concentration of 5 X 10(5) cells/well, zymosan-activated serum (ZAS), or autologous serum or plasma as the chemoattractants, and a 120-minute incubation period. Canine neutrophils responded well to ZAS, heat-inactivated ZAS, autologous serum and plasma, and heat-inactivated pooled serum. Chemotactic activity was proportional to the concentration of serum used as the chemoattractant. Mean (+/- SD) random migration, chemotaxis, chemotactic index, and chemotactic differential of neutrophils from 9 healthy Greyhounds were 1.09 (+/- 0.23), 1.95 (+/- 0.38), 1.82 (+/- 0.31), and 0.86 (+/- 0.32) mm,respectively.

Three trials were conducted to establish if young primary specific pathogen free (SPF) pigs could be protected from Glasser's disease by vaccination. Three age groups of cesarean-derived isolator-reared gnotobiotic pigs were vaccinated twice at 4 and 6, 3 and 5, and 2 and 4 wk of age respectively with a formalin killed aluminum hydroxide adsorbed bacterin prepared from three strains of Haemophilus parasuis isolated from Ontario pigs affected with Glasser's disease. When challenged two weeks later with the homologous strains of virulent bacteria, all the vaccinated pigs remained healthy, while 17/18 nonvaccinated pigs became severely sick or died between three and seven days postchallenge. The one surviving nonimmunized pig was retarded in growth. All of the nonimmunized pigs had visible lesions of polyserositis, the most common being polyarthritis (14/18). Other lesions were ribrinous meningitis, pericarditis, pleurisy and/or peritonitis. Two of the pigs died with a septicemia. Haemophilus parasuis was isolated from 15/18 nonimmunized pigs, usually from several of the affected sites. The organisms were not isolated from the immunized pigs, nor from the surviving nonimmunized pig. Attempts to detect the presence of specific antibodies against the H. parasuis strains in the sera of the immunized or exposed pigs by the passive hemagglutination test or by enzyme linked immunoassay were unsuccessful. The results of this work indicate that primary SPF pigs can be protected from Glasser's disease by vaccination as early as 2 and 4 wk of age. The nature of this protective mechanism was not established in this study.

An automated reticulocyte counting method that used a flow cytometer and the nucleic acid staining dye, thiazole orange, was developed. Anticoagulated (EDTA) blood specimens were suitable for flow cytometric reticulocyte counting when stored at 4 C for 96 hours after collection. Thiazole orange-stained samples were stable for 5.5 hours after staining when stored capped at 20 C and protected from light. Flow cytometric and manual microscopic reticulocyte counts were compared for counts in the 0.27 to 5.32% range (as determined by flow cytometry) and 0.10 to 4.90% range (as determined by 1 technician). Although the results of flow cytometric analysis generally correlated well (r = 0. 821) with manual counts, there was poor correlation between the procedures for counts less than or equal to 2.0% (r < 0.272). Linearity of flow cytometric counts over the range 0.27 to 14.46% was excellent (r = 0.999). Within-run precision of flow cytometric counts (% coefficient of variation [CV] = 3 to 5) was superior to manual microscopic counts obtained by one technician (% CV = 19 to 23) and to manual microscopic counts, which were an average of counts done by 3 technicians (% CV = 8 to 18). Comparable flow cytometric counts were obtained by counting 50,000 or 100,000 blood cells in the flow cytometer.

The effects of a potent new histamine-2 (H2) receptor antagonist, BMY-25368, were studied on gastric acid secretion in 5 foals from which food was withheld. Doses of 0.02, 0.11, 0.22, and 1.10 mg/kg of body weight were administered IM in a randomly assigned treatment sequence. Following BMY-25368 administration, hydrogen ion concentration was decreased and mean pH was higher than baseline values in a dose-response pattern. At the 0.22 and 1.10 mg/kg doses, the high pH was sustained for > 4 hours. The BMY-25368 thus may be useful for treating gastric ulcer disease in horses.

Total protein concentration was determined in serum, bronchoalveolar lavage (BAL) fluid, and nasal flush fluid obtained from specific-pathogen-free cats from birth to maturity and from adult conventionally raised cats. Protein components were analyzed by immunoelectrophoresis and isoelectric focusing. Albumin, and alpha, beta, and gamma-globulins were among the proteins identified in BAL fluid, and their isoelectric point ranged from 3.1 to 5.1. gamma-Globulin was not detected in serum or BAL fluid of newborn kittens before they had ingested colostrum. By day 3 after ingestion of colostrum, IgG was detected in high concentration in serum and was the predominant immunoglobulin in serum and BAL fluid of older cats. Nasal flush fluid from cats > 6 months old contained albumin, and alpha, beta, and gamma-globulins, with IgA being the predominant immunoglobulin. Total protein concentration in nasal flush fluid increased progressively with increasing age, and albumin was the predominant protein. Protein concentration was significantly (P < 0.01) higher in BAL fluid from conventionally raised adult cats than in that from specific-pathogen-free cats.

Bovine whey samples were evaluated by use of lymphocyte-transformation tests to determine their effect on lymphocyte blastogenesis. Whey samples from mammary glands with clinical mastitis strongly inhibited DNA synthesis and blastogenesis in lymphocytes stimulated with mitogens or dividing because of bovine leukemia virus infection. Whey samples from apparently healthy glands either did not inhibit lymphocyte DNA synthesis or inhibited it to a lesser degree than did whey from mastitic glands. Degree of inhibition was dose-dependent. The molecules causing inhibition were noncytotoxic and underwent minimal binding to the lymphocytes. Inhibitory molecules were susceptible to various proteolytic and glycolytic enzymes, indicating a glycoprotein-like structure. Whey inhibited incorporation of thymidine if it was in the cell cultures during the early stages of stimulation. Incubation of lymphocytes in whey that inhibited thymidine incorporation did not affect DNA synthesis in subsequent culturing of the same cells without whey. Degree of inhibition was affected by the method of whey preparation.

Evaluation of urethral pressure profilometry (UPP) with simultaneous fine-wire electromyography of the external urethral sphincter (EUS) was conducted in 11 healthy adult male cats sedated with xylazine and ketamine. A 3.5-F urethral catheter with a closed end and two 1-mm sideports was infused with sterile 0.9% NaCl solution at a rate of 2 to 3 ml/min. A fine-wire electromyographic (EMG) electrode was placed percutaneously into or near the external urethral sphincter prior to the onset of the UPP. The maximal urethral pressure achieved and functional profile length were recorded from UPP. Setting both catheter withdrawal rate and paper speed at 5 mm/s enabled the measurement of actual urethral length directly from UPP. Sphincter EMG activity was rated as slight (+), moderate (+ +), or intense (+ + +). All recordings were replicated once during each trial for 8 cats and trials were replicated 5 to 7 days later in 4 cats. Before catheterization, EMG activity of the external urethral sphincter was rated slight (+), whereas intense (+ + +) activity accompanied insertion. The activity evoked by movement of the catheter subsided, but intense EMG activity of the external urethral sphincter was recorded from onset to completion of catheter withdrawal in all cats in both trials. The mean maximal urethral pressure was 93.1 +/- 13.29 cm H2O. The mean function urethral length was 8.1 +/- 0.93 cm. Maximal urethral pressure or function profile length did not differ significantly between recordings within trials or between trials. Simultaneous recording of EMG activity and UPP of the external urethral sphincter was shown to be a simple, noninvasive technique for assessing neuromuscular and anatomic urethral function.

The pharmacokinetic properties of indomethacin and its effects on aqueous protein values were studied in 15 clinically normal Beagles. The dogs were treated every 6 hours with 1% indomethacin suspension in 1 eye, with the other eye serving as a control. After 24 hours, the dogs were anesthetized and samples of aqueous humor (AH) were drawn by aqueocentesis at 0, 15, 30, 60, and 90 minutes after initial paracentesis. Additional samples were drawn at the time of euthanasia, 180 (6 dogs) and 360 minutes (9 dogs) minutes after initial paracentesis. Blood samples were obtained at each treatment and at each aqueocentesis. The eyes were enucleated after dogs were euthanatized. Aqueous protein concentrations and indomethacin concentrations in AH, plasma, and different ocular tissues were determined. Topical indomethacin administration had no effect on baseline protein concentrations of AH. It reduced protein concentrations in AH significantly at all times after initial aqueocentesis. This reduction was approximately 30%. Indomethacin in the AH is mostly protein-bound. Concentrations were 350 ng/ml in primary AH and 1,305 ng/ml in secondary AH, 90 minutes after initial aqueocentesis. Free-drug concentrations were relatively constant at about 220 ng/ml. Indomethacin administered topically is readily absorbed by the ocular adnexae, reaching a steady-state concentration of 25 ng/ml in blood plasma 18 hours after the start of treatment. Plasma concentrations were 50 times lower than therapeutically effective concentrations. High indomethacin concentrations were found in the cornea only. Low concentrations were found in the iris and ciliary body, the lens, and in the choroid. On the basis of our findings, we conclude that topically administered indomethacin is effective in reducing protein concentrations in secondary AH and is rapidly eliminated from the eye.