The popular urodynamic technique of stressed urethral pressure profilometry used for investigation of genuine stress incontinence in women was adapted and applied to bitches. The aim was to assess the suitability and reproducibility of the technique in the canine species, and to determine whether differences seen in continent and incontinent women were found in bitches. Resting and stressed simultaneous urethral pressure profiles were obtained for 25 continent and 25 incontinent bitches, the latter diagnosed as having urethral sphincter mechanism incompetence. The stressed urethral pressure profiles were produced by ballottement of the abdomen during catheter withdrawal. The degree of stress induced was consistent and had got short-term reproducibility. Highly significant (P < 0.001) differences in the percentage of negative spikes extending below the resting intravesical pressure were found between continent and incontinent bitches. Significant differences were not observed in the pressure transmission profiles between continent and incontinent bitches; both groups had a gradual decrease in pressure transmission from the bladder neck to the external urethral orifice. The distance from the start of the urethral pressure profile to the first negative peak (attributable to respiration or ballottement) on the subtracted profile was compared with the radiographic distance that the bladder neck was positioned with respect to the cranial pubic brim, taking body weight and continence status into account. Body weight and continence status did not have significant effect on the relation in either instance. The distance between the start of the urethral pressure profile and the first negative peak induced by respiration was significantly (P < 0.05) related to the bladder neck position with respect to the cranial pubic brim, although it accounted for little of the total variance. Relation between the same variables during stressed urethral pressure profilometry, induced by abdominal ballottement, was not significant.

Four-week-old chickens were inoculated orally with 1,000 or 100,000 oocysts of the ME-49 or GT-1 strain of Toxoplasma gondii, and their antibody responses were measured, using the direct modified agglutination test, latex agglutination test, indirect hemagglutination test, ELISA, and the Sabin-Feldman dye test. Antibodies against T gondii were detected by use of the modified agglutination test and ELISA within 2 weeks of oocyst inoculation, and antibodies persisted until termination of the study by postinoculation day 68. The latex agglutination test was insensitive in detecting T gondii antibodies, and antibodies were not detected by use of the dye and indirect hemagglutination tests. Of tissues bioassayed in mice for tissue cysts by pepsin digestion of individual organs of chickens on postinoculation day 68, tissue cysts were found in the brain of all 5, heart of 3, and leg muscles of 2, but not in the liver and breast muscles. None of the birds developed clinical toxoplasmosis.

Each of 5 US-origin serotypes of bluetongue virus (BTV) was inoculated into a separate pair of sheep. The duration of each animal's ensuing viremia was monitored, using a BTV serogroup-specific nested polymerase chain reaction (PCR) method and an embryonating chicken egg (ECE) inoculation procedure. Mean duration of viremia was 100 and 38 days for the PCR and ECE methods, respectively. This difference was significant (P < 0.001) and documents a more prolonged viremia in virus-exposed sheep than has been reported. A dual internal oligonucleotide solution hybridization procedure was developed for the rapid (2 hours) colorimetric detection and identification of BTV-specific PCR products. This enzyme-linked oligonucleotide sorbent assay (ELOSA) relied on annealing of separate biotinylated and fluoresceinated probes to the amplified BTV nucleic acid; these complexes were captured on streptavidin-coated microtitration wells and were detected, using a horseradish peroxidase-labeled antifluorescein antibody conjugate. End-point dilution analyses of PCR products indicated that the ELOSA was more sensitive than gel electrophoretic or comparable colorimetric slot-blot hybridization techniques. The BTV PCR-ELOSA system represents a more sensitive and expeditious means of diagnosing BTV-induced viremia than does the ECE procedure currently used. The combination of ELOSA with PCR should facilitate practical application of nucleic acid technology to diagnostic veterinary medicine.

Pyelonephritis was experimentally induced in 10 clinically normal dogs by nephropyelocentesis and introduction of Proteus mirabilis into the randomly chosen right or left renal pelvis. Dogs were examined by nephrosonography and excretory urography before and 2 weeks after infection. The major nephrosonographic findings of pyelonephritis were renal pelvic dilatation, usually with proximal ureteral dilatation, and a hyperechoic mucosal margin line within the renal pelvis, proximal portion of the ureter, or both. In addition, at least one or more of the following were observed: generalized hyperechoic renal cortex, focal hyperechoic areas within the medulla, and focal hyperechoic or hypoechoic cortical lesions. Interpretation of excretory urograms resulted in 3 false-negative and 1 false-positive conclusions, compared with the histologic findings. Interpretation of nephrosonograms resulted in 2 false-negative and no false-positive conclusions. Of the kidneys with histologic evidence of pyelonephritis, 73% were detected by excretory urography, whereas 82% were detected by nephrosonography. Nephrosonography appeared to be useful for detection of mild to moderate cases of acute pyelonephritis that may be be interpreted as such by excretory urography.

Flow-volume loops generated from 6 Standardbreds at rest and during treadmill exercise were evaluated for their use in detecting upper airway obstruction. Tidal breathing flow-volume loops (TBFVL) were obtained from horses at rest and exercising at speeds corresponding to 75% of maximal heart rate and at maximal heart rate. The TBFVL were evaluated, using a pulmonary function computer; calculated indices describing airflow rate and expiratory-to-inspiratory airflow ratio for individual loops were determined. In addition to TBFVL indices, standard variables of upper airway function also were measured: peak airflow, peak pressure, and calculated inspiratory and expiratory impedances. Measurements were recorded before left recurrent laryngeal neurectomy (LRLN; baseline) and 14 days after surgically induced left laryngeal hemiplegia. When horses were at rest, TBFVL shape and indices describing the loop were highly variable. In contrast, in exercising horses, TBFVL shape was consistent and coefficients of variation of loop indices were less during exercise than at rest. After LRLN, TBFVL from exercising horses indicated marked inspiratory airflow limitation, while the expiratory airflow curve was preserved. Peak inspiratory flow rate and inspiratory flow at 50 and 25% of tidal volume decreased, and the ratio of peak expiratory to inspiratory airflow and that of midtidal volume expiratory and inspiratory airflow rates increased significantly (P < 0.05). Inspiratory impedance also increased after LRLN. Although in resting horses TBFVL were not a useful indicator of upper airway obstruction, examination of TBFVL from exercising horses allowed objective, specific, and repeatable detection of upper airway obstruction. The technique was noninvasive, rapid, and well tolerated by horses; thus, it is a potentially valuable clinical diagnostic test.

A specific polymerase chain reaction (PCR) assay was devised, allowing detection of 1 bovine leukemia virus (BLV)-infected cell in 10(4) bovine lymphocytes. The efficacy of field application of the developed method was verified by evaluating the rate of viral transmission to calves from infected cows, whether they have persistent lymphocytosis. With this objective, 43 calves were simultaneously tested at birth and at 6 months of age for viral antibodies in serum and for proviral DNA in lymphocytes. At birth, 36 calves were BLV-negative and 3 were BLV-positive by results of serologic and DNA-based assays. Conversely, results for 4 calves had lack of correlation between the diagnostic methods. In particular, 2 calves were DNA-positive and antibody-negative for BLV and 2 other calves had the opposite test results. At 6 months of age, when the immunologic pattern more closely reflects the status of calves' immune response, independent of maternal antibodies, all calves DNA-negative for BLV at birth (n = 38), were consistently PCR- and antibody-negative for BLV. On the contrary, the cattle DNA-positive for BLV at birth (n = 5), whether seropositive or not, were PCR- and antibody-positive for BLV, at the time of the second screening. Thus, these results indicate reliability of the PCR to diagnose perinatal BLV infection. Furthermore, the observation that all calves found to be infected at birth were born to BLV-positive cows with persistent lymphocytosis, indicates that the persistent lymphocytosis status of the cow may represent a factor associated with BLV infection in utero.

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.

Undifferentiated distal respiratory tract disease (nasal discharge, cough, pneumonia) in foals (1 to 8 months old) is a burdensome economic problem on breeding farms yet, the infective agents associated with these episodes have not been well described. Possible causes of these episodes of illness were investigated by culturing specimens of proximal and distal airways of clinically diseased foals (n = 101), prior to any treatment, for aerobic and anaerobic bacteria and viruses (rhinoviruses, equine arteritis virus, equine herpesvirus subtype 1 [EHV-1], influenza virus, and adenovirus). Pairs of sera (n = 47) were examined for antibodies to influenza A virus, equine subtypes 1 and 2, EHV-1, and adenovirus antigens, and sera obtained from foals during acute infection were examined for antibodies (by agar gel immunodiffusion [AGID]) to equi factor antigens of Rhodococcus equi. Viruses were not isolated from the proximal (swab) or distal (bronchial lavage) airway specimens in foals, and only 2 of 47 randomly selected foals seroconverted to EHV-1. Serotiters to the other viruses were low and frequently decreasing between samples, which was compatible with maternally derived antibody. Streptococcus zooepidemicus was the predominant isolate from bronchial lavage specimens (88/101 cases), accompanied by alpha-hemolytic streptococci (8 cases), Bordetella bronchiseptica (13 cases), Staphylococcus epidermidis (9 cases), and other organisms in lesser frequency. Only Str zooepidemicus was recovered significantly (P < 0.05) more often in cases than in controls. The AGID test was found useful to detect 1/26 with presumed exposure to R equi, but positive tests results did not correspond well with bacterial culture results; positive AGID results were recorded in 34% of culture-negative foals. However, foals from which R equi was isolated were distinctive from the other foals on the basis of fever (> 39 C), lack of nasal discharge, blood neutrophilia and decreased percentage of neutrophils in bronchial lavage fluid samples. Isolation of Str zooepidemicus was significantly (P < 0.01) associated with increasing neutrophil percentage in bronchial lavage fluid. In conclusion, the pathogenic roles of Str zooepidemicus and R equi were established in this group of foals with distal respiratory tract infections by use of clinical, endoscopic, hematologic, and cytologic methods. There was no evidence of a viral cause for these infections, indicating that manifestations of distal respiratory tract infection are attributable to bacterial infection causing inflammation of the airways. Further studies are warranted to pursue more-sensitive methods for detection of viral antigen or antibody in undifferentiated distal respiratory tract disease episodes in foals.

A sensitive and specific DNA probe for detection and identification of bovine herpesvirus 4 (BHV-4) was developed. Cloned fragments from a library of HindIII fragments of the BHV-4 (DN-599) genome were labeled with 32P or digoxigenin and were tested for sentitivity and specificity in detecting viral DNA by dot-blot hybridization. Two probes were identified that detected 10 pg of purified viral DNA, and detected viral DNA in 0.001 microgram of total DNA extracted from BHV-4-infected cells. Both probes labeled with 32P and 1 labeled with digoxigenin detected viral DNA in samples prepared from cells infected with 2 prototype strains (DN-599 and Movar 33/63) and 4 field isolates of BHV-4. The DNA probes did not hybridize to total DNA prepared from uninfected bovine cells or from cells infected with BHV-1, BHV-2, alcelaphine herpesvirus 1, pseudorabies virus, or equine herpesvirus 1. One probe, labeled with digoxigenin, was tested further by dot-blot hybridization with infected cell lysates that were simply treated with sodium dodecyl sulfate and proteinase K prior to application to the membrane, avoiding extensive DNA purification procedures. This simplified procedure also resulted in specific detection of field isolates of BHV-4 and prototype strains of BHV-4.