In an abattoir-based case-control study, histologic, and macroscopic examination of porcine intestines at slaughter and 2 molecular assays were compared for use as diagnostic tests of proliferative enteritis (PE). Fecal samples and intestinal specimens were collected from pigs with grossly thick ileum and from clinically normal pigs at slaughter. Tissue specimens were fixed in neutral buffered 10% formalin, and sectioned. Sections stained with H&E were examined for proliferative lesions by a pathologist unaware of the group to which the pig had been assigned on the basis of results of gross examination. Adjacent tissue sections, stained with Warthin-Starry (silver) stain, were examined for presence of the intracellular bacterium of PE, ileal symbiont (IS)-intracellularis, in the enterocytes of the intestinal crypts by the senior author, who was unaware either of the group to which the pig had been assigned or diagnosis by the pathologist. Bacterial DNA was extracted from the fecal samples and assayed by dot-blot hybridization and polymerase chain reaction (PCR) for presence of IS-intracellularis DNA, without knowledge of results of the other examinations. The PCR assay for IS-intracellularis was a specific and sensitive diagnostic test for PE, and dot-blot hybridization was sensitive, but was less specific. Macroscopic examination of intestines at slaughter was a sensitive, but not specific, test. Association between IS-intracellularis and proliferative lesions was statistically examined in the same study. There was a highly significant (P = 0.0078) association between presence of naturally acquired proliferative lesions and intracellular infection induced by IS-intracellularis. The odds ratio of greater than or equal to 14 and estimated attributable fraction of greater than or equal to 92% indicate that IS-intracellularis may be a necessary cause of PE.

The ability of viral glycoproteins (VP) VP4/VP7 reassortant swine rotaviruses (RV) to induce cross-neutralizing antibody against parental serotypes was investigated in guinea pigs. Using selective culture conditions, we produced 10 reassortant viruses that contained gene segment 4 of the OSU RV strain and gene segment 9 of the Gottfried RV strain. These reassortant RV grew to high titer in cell culture and were neutralized by monospecific antisera against both parental RV strains. The reassortant RV were chemically inactivated with binary ethylenimine, adjuvanted with aluminum hydroxide, and used to produce antisera in guinea pigs. The hyperimmune antisera had high neutralization titer against both parent RV strains. These results indicate that several of the reassortant RV may be capable of inducing neutralizing antibodies to VP4 and VP7 and may have future use as bivalent vaccine strains.

Gastric pH was monitored in neonatal foals from birth to 3 months of age. Background pH decreased, especially during the first week of life. Milk had complex effects that depended on pH prior to sucking, confounded by the age of the foal: nearly neutral background pH tended to be acidified after milk intake; moderately acid background pH tended to be neutralized; low background pH was only slightly increased by milk. Absolute magnitude of the effects of milk decreased with age. Existence of a proulcerative intragastric environment in preweaning foals is postulated, but this must be considered in the context of what probably is a multifactorial pathogenesis.

A 3-year prospective study of large-breed dogs (4 months to 3 years of age) was conducted to evaluate the influence of radiographic positioning and age on coxofemoral joint (hip) laxity, subjective hip score, and development of degenerative joint disease (DJD). The dogs (n = 142) were breeder- or client-owned and represented 14 breeds. With dogs under heavy sedation, hips were radiographed in the standard hip-extended position and in the new compression/distraction position at 4, 6, 12, 24, and 36 months of age. The standard hip-extended radiographic view was evaluated by 3 methods: subjective evaluation by a board-certified veterinary radiologist (WHR), according to the standard 7-point Orthopedic Foundation for Animals (OFA) scoring scheme (OFA/WHR); joint laxity quantitation, using the Norberg angle (NA) method; and subjective scoring by a veterinary orthopedic surgeon for radiographic evidence of DJD. The hips in the distraction radiographic view were evaluated for passive hip laxity, as measured by use of a unitless distraction index (DI). Results of the study indicated that at a specific age (4, 6, 12, 24, or 36 months), all methods of hip evaluation correlated with each other at a moderate level (P < 0.05). The strength of contemporaneous correlation tended to increase with age of evaluation. Longitudinally, the between-method correlations were usually significant (P < 0.05), but not at a sufficiently high level to permit reliable between-method prediction. Prospective intraclass (within-method) statistical analysis of the various hip-scoring methods indicated that DI was superior to NA and OFA/WHR in comparability of score over time. The intraclass correlation coefficient ranged from 0.55 to 0.91 for DI in contrast to 0.40 to 0.78 for NA, and 0.06 to 0.39 for OFA/WHR over the age intervals of the study. For reference, the highest Kappa of 0.39 for the subjective OFA/WHR scoring reflected a maximal level of agreement between time intervals, only slightly better than chance. The associated large error questions the predictive use of the 7-point, subjective hip-scoring scheme, particularly prior to the age of 2 years.

Two distinct monoclonal antibodies (MAB) were prepared for testing with kidney, spleen, and retrobular tissue imprints made from chinook salmon (Oncorhynchus tshawytscha) affected with plasmacytoid leukemia (PL). Hybridomas were prepared from mice immunized with whole and lysed cells purified from renal or retrobular PL-positive tissues, which had been obtained from naturally and experimentally infected fish from British Columbia, Canada. The MAB reacted with at least 4 morphologically different cell types; fluorescence was associated with the plasma membrane and cytoplasm. The MAB also reacted with kidney imprints made from chinook salmon affected with a PL-like lymphoproliferative disease in California, indicating that these 2 diseases might be caused by a similar agent. The MAB did not react with any of the kidney or spleen imprints made from wild chinook salmon collected from a river in Ontario, Canada.

Intragastric pH monitoring was investigated in ponies. In cadaver stomachs, close contact with the mucosa led to high pH readings if nonweighted electrodes were used. However, pH recorded by weighted electrodes was markedly less affected by mucosal contact (P < 0.001). The latter were used for subsequent trials. In vivo, high correlations were found between pH recorded by weighted electrodes with or without a wire guard to prevent mucosal contact (correlation, r = 0.866; P < 0.001). Readings from each correlated well with those from simultaneous gastric aspirates (r = 0.774 and r = 0.807, respectively; P < 0.001 for both correlations). Plain electrodes recorded more highly variable (temporally heterogeneous) pH than did guarded electrodes. In vitro, trials using equine gastric fluid indicated that this resulted from greater responsiveness of the plain electrode. In vivo, episodes of nearly neutral pH were a common feature, and high pH correlated with intensely yellow-green, neutral fluid in the stomach (rank correlation, p = 0.626; P < 0.01). Concentration of bile acids did not correlate with pH or color score (p = -0.158 and p = 0.076, respectively). Causes of the episodes could include salivary influx, duodenogastric reflux, and variable gastric acid secretion. Pentagastrin infusion (0.6 micrograms/kg of body weight/ h) reduced intragastric pH (P = 0.018), but episodes of neutrality still occurred. Experiments in fed ponies indicated possible existence of a stable pH gradient, from neutral dorsally to heterogeneous and more acidic ventrally. Care was required in the rational choice of summary variables for expression of monitored pH data. Of the frequency distributions of 3 summary variables assessed in this study (mean, median, and percentage of data > pH 4), only that of the mean approached normality. Thus, use of the mean may allow analysis by parametric statistical methods. Intragastric pH monitoring was found to be a useful technique. Episodes of increased pH were often identified. These may represent episodic duodenogastric reflux.

Bordetella avium is an important respiratory tract pathogen of turkeys. In common with other pathogenic bordetellae, B avium manifests a tissue tropism for cilia of the respiratory tract epithelium. To determine the molecular characteristics of the host cell receptors for B avium, we used hemagglutination and in vivo adherence assays. Carbohydrates, mucus, sialic acid-specific lectin, and other glycoconjugates were evaluated for their ability to competitively inhibit binding of B avium to host cells. The gangliosides, GD(1a) and GT(1b), completely inhibited hemagglutination, whereas N-acetylneuraminic acid (sialic acid) partially inhibited hemagglutination. Adherence to turkey tracheal mucosa in vivo was significantly (P < 0.01) inhibited by GD(1a) and GT(1b) gangliosides, N-acetylneuraminic acid, bovine sub-maxillary mucin, and horseshoe crab (Limulus polyphemus) lectin. Treatment of the tracheal mucosa with neuraminidase also inhibited adherence of B avium. We conclude that N-acetylneuraminic acid and the gangliosides, GD(1a) and GT(1b) may be important components of the tracheal mucosa receptor for B avium in turkeys.

Procedures were developed to count neutrophils in milk using a flow cytometer. Milk samples from 2 experiments were counted: 1 with 4 noninfected cows and a second with 5 noninfected cows that were injected with endotoxin in 2 mammary quarters. Thus, the procedures were evaluated on normal milk and on that with high somatic cell count. Flow cytometric procedures involved fluorescence detection (from the dye carboxydimethylfluorescein diacetate) to distinguish intact and viable from fragmented cells, forward light scatter to detect cell size differences, and right-angle side scatter to detect cellular granularity. High fluorescence, large size, and high degree of granularity identified viable neutrophils. For all samples, neutrophils were also counted manually, using the cytologic centrifugation approach to create the slides; manual counts were used as the standard for comparison. In experiment 1 (normal milk), mean values for percentage of viable neutrophils estimated by manual and flow cytometry procedures agreed closely (26% vs 25.8% for foremilk and 28.8% vs 26.6% for bucket milk). Sources of variation in manual and flow cytometric estimates of percentage of neutrophils were examined. Cow variation was significant (P < 0.01) for manual and flow cytometric counts, but was larger for flow cytometric counts. Day-to-day variation in counts on milk from the same cow was negligible for manual counts, but was significant (P < 0.01) for flow cytometric counts. Coefficients of variation were considerably Larger for manual counts than for flow cytometry. In experiment 2 (milk with high cell count, foremilk), agreement between mean values obtained by flow cytometry and by manual counting was somewhat less. However, predicting manual percentage of neutrophils, using the flow cytometric estimate, had R2 of 0.77. Regression of the manual percentage of neutrophils value on the flow cytometric percentage of neutrophil value was close to 1.0, with only a small negative intercept. Some additional refinement of flow cytometric procedures may be required before flow cytometric estimates of percentage of neutrophils can be accepted without simultaneous validation by use of manual counting. In particular, causes of day-to-day variation in flow cytometric results should be identified and reduced.

The prevalence of feline leukemia virus (FeLV) antigen and DNA was assessed in formalin-fixed, paraffin-embedded tumor tissues from 70 cats with lymphosarcoma (LSA). Tissue sections were tested for FeLV gp70 antigen using avidinbiotin complex (ABC) immunohistochemistry (IHC); DNA was extracted and purified from the same tissue blocks for polymerase chain reaction (PCR) amplification of a 166 base pair region of the FeLV long terminal repeat (LTR). Results were related to antemortem FeLV enzyme-linked immunosorbent assay (ELISA) for serum p27 antigen, anatomic site of LSA, and patient age. Viral DNA was detected by PCR in 80% of cases and viral antigen by IHC in 57% of cases. Seventeen cases were PCR-positive and IHC-negative; one case was PCR-negative and IHC-positive. Clinical records included FeLV ELISA results for 30 of 70 cats. All 19 ELISA-positive cats were positive by PCR and IHC; of the 11 ELISA-negative cats that were negative by IHC, seven were positive by PCR. When evaluated according to anatomic site, FeLV DNA and antigen were detected less frequently in intestinal LSAs than in multicentric and mediastinal tumors. Lymphosarcoma tissues from cats < 7 yr were several fold more likely to be positive for FeLV antigen by IHC than were tumors from cats > or = 7 yr. However, there was no significant difference in PCR detection of FeLV provirus between LSAs from cats < 7 yr and those > or = 7 yr. These proviruspositive, antigen-negative cases may represent infection with latent or replication-defective FeLV.

Results of an ELISA, indirect fluorescent antibody (IFA) test, and immunoblot analysis (western blotting) for antibody to Borrelia burgdorferi in a sample of 216 lactating dairy cows were compared. The microscopic microtitration agglutination test for antibody to 6 serovars of Leptospira interrogans was also performed to evaluate possible cross-reactivity between B burgdorferi and L interrogans. Using western blotting as the standard test against which the ELISA and IFA test were compared, the ELISA had greater sensitivity (50% in summer and 38% in spring) with similar specificity (83 and 82%), compared with the IFA test (sensitivity, 6 and 5%; specificity, 90 and 83%). In addition, seropositivity to B burgdorferi, using the ELISA, was not found to be associated with seropositivity to L interrogans serovars. A matched case-control study evaluating the association between clinical lameness and antibody to B burgdorferi was performed in lactating dairy cows of 17 Minnesota and Wisconsin herds. Sera from case and control cows matched by herd, parity, and stage of lactation were evaluated, using an ELISA for B burgdorferi antibody during 2 seasons. High B burgdorferi antibody values were associated with clinical lameness in dairy cows (P = 0.006 in summer and P = 0.04 in spring).