OBJECTIVE To assess agreement between 2 benchtop blood gas analyzers developed by 1 manufacturer (BGA 1 and BGA 2 [a newer model with reduced maintenance requirements]) and a reference chemistry analyzer for measurement of electrolyte (sodium, chloride, and potassium) in blood samples from dogs. ANIMALS 17 healthy staff- and student-owned dogs and 23 client-owned dogs admitted to an emergency and intensive care service. PROCEDURES Blood collected by venipuncture was placed in lithium heparin–containing tubes. Aliquots were analyzed immediately with each BGA. Samples were centrifuged, and plasma was analyzed with the reference analyzer. Results for each BGA were compared with results for the reference analyzer by Passing-Bablok regression analysis. Percentage differences between BGA and reference analyzer results were compared with published guidelines for total allowable error. RESULTS Proportional bias was detected for measurement of chloride concentration (slope, 0.7; 95% CI, 0.7 to 0.8), and constant positive bias was detected for measurement of chloride (y-intercept, 34, mmol/L; 95% CI, 16.9 to 38 mmol/L) and potassium (y-intercept, 0.1 mmol/L; 95% CI, 0.1 to 0.2 mmol/L) concentrations with BGA 1. There was no significant bias for measurement of potassium or chloride concentration with BGA 2 or sodium concentration with either BGA. Differences from the reference analyzer result exceeded total allowable error guidelines for ≥ 1 sample/analyte/BGA, but median observed measurement differences between each BGA and the reference analyzer did not. CONCLUSIONS AND CLINICAL RELEVANCE Good agreement with reference analyzer results was found for measurement of the selected electrolyte concentrations in canine blood samples with each BGA.

To determine whether an anti-Salmonella bacterium is involved in control of pathogen load in persistently infected cattle herds. 24 Holstein calves experimentally infected and 39 Holstein cows naturally infected with Salmonella spp. An Escherichia coli (designated as P8E5) that possessed anti-Salmonella activity was isolated from Salmonella-negative bovine feces obtained from a herd with endemic Salmonella infection. In vitro analysis involved enumerating Salmonella enterica serovar Typhimurium coincubated with E coli P8E5. In vivo analysis involved coadministration of Salmonella spp and E coli P8E5 or an E coli control strain to neonatal Holstein calves. Fecal samples were collected on multiple days after inoculation, and quantitative PCR assay was performed by use of Salmonella-specific primers. E coli P8E5 reduced viability of Salmonella spp in vitro. Shedding of Salmonella organisms was diminished in calves administered E coli P8E5, whereas the control strain of E coli had no effect on shedding of Salmonella organisms. In this study, an E coli strain was identified that possessed bacteriocin-like activity and was able to decrease viability of Salmonella organisms in vitro and in vivo. Therefore, it is possible that this organism could be representative of native microbiota that dampen Salmonella spp in persistently infected cattle herds.

Objective-To analyze the 7a7b genes of the feline coronavirus (FCoV) of cheetahs, which are believed to play a role in virulence of this virus. Sample Population-Biologic samples collected during a 4-year period from 5 cheetahs at the same institution and at 1 time point from 4 cheetahs at different institutions. Procedures-Samples were first screened for FCoV via a reverse transcription-PCR procedure involving primers that encompassed the 3'-untranslated region. Samples that yielded positive assay results were analyzed by use of primers that targeted the 7a7b open reading frames. The nucleotide sequences of the 7a7b amplification products were determined and analyzed. Results-In most isolates, substantial deletional mutations in the 7a gene were detected that would result in aberrant or no expression of the 7a product because of altered reading frames. Although the 7b gene was also found to contain mutations, these were primarily point mutations resulting in minor amino acid changes. The coronavirus associated with 1 cheetah with feline infectious peritonitis had intact 7a and 7b genes. Conclusions and Clinical Relevance-The data suggest that mutations arise readily in the 7a region and may remain stable in FCoV of cheetahs. In contrast, an intact 7b gene may be necessary for in vivo virus infection and replication. Persistent infection with FCoV in a cheetah population results in continued virus circulation and may lead to a quasispecies of virus variants.

The role of interleukin 1 (IL-1) as an inflammatory mediator during mastitis and the therapeutic effect of recombinant human IL-1 receptor antagonist (IL-1ra) for bovine mastitis was studied. Cows were intramammarily infused with lipopolysaccharide (25 g) in 1 mammary gland. Half the cows also received infusions of 5 mg of IL-1ra into the same mammary gland just prior to endotoxin infusion and 4, 8, and 12 hours later. After endotoxin infusion, tumor necrosis factor and high IL-1 bioactivity were detected in whey from infused glands. Vascular permeability changes and neutrophil accumulation in milk paralleled the appearance of cytokines. A systemic reaction, characterized by pyrexia and an increase in blood cortisol concentration, also were observed. Milk yield was inhibited and milk composition was altered in infused and noninfused glands. The increase in IL-1 bioactivity in milk after endotoxin infusion was almost completely prevented in glands receiving IL-1ra. However, IL-1ra had no effect on local inflammation, systemic reaction, or impairment in productive performance. These results indicate that IL-1 does not mediate its effect within the milk compartment, and suggest either that IL-1 is not critical to the mastitic response or that intramammary infusion of IL-1ra does not place the antagonist where IL-1 interacts with its receptor.

We investigated the effect of adding psyllium to a standard electrolyte solution in 10 calves with diarrhea. The calves were tested with the standard solution on one day and standard solution plus psyllium on the alternate day. The order of treatments was randomized. Psyllium converted the solution into mucilage, but did not affect fecal consistency. Mean +/- SEM area under the glucose absorption curve was lower for mucilaginous than for nonmucilaginous solutions, 2.1 +/- 0.62 vs 3.75 +/- 1.18 mmol.h, respectively, but the difference was not significant. The area under the breath hydrogen curve was marginally lower for mucilaginous than nonmucilaginous solutions, 102 +/- 20 and 209 +/- 60 ppm.h, respectively. The usefulness of such decreased bacterial fermentation is doubtful.

Interstitial and bronchointerstitial pulmonary patterns are commonly observed in thoracic radiographs of Thoroughbreds. Prominent interstitial and bronchointerstitial pulmonary patterns are observed in clinically normal horses, and in horses with respiratory tract disease. Until recently, the relevance of these pulmonary patterns was not known. Previous studies indicated that bronchiolitis, bronchiolar epithelial hyperplasia, epithelial metaplasia, and bronchial arteriolar recruitment correlated strongly with the prominence of the interstitial and bronchointerstitial pulmonary patterns observed radiographically. We examined the content and distribution of collagen in the lungs of 7 clinically normal Thoroughbreds in race training. After standardized fixation, lung tissue was treated with a compound that selectively stains collagen. Standard morphometric techniques were used to determine the volume density of parenchymal tissue and parenchymal airspace, mean linear intercept (estimate of alveolar size), alveolar surface area-to-volume ratio, percentage of parenchyma composed of collagen, percentage of airway wall composed of collagen, and airway wall thickness. These values were compared with radiographic and histopathologic scores obtained from the same horses. The volume density of parenchymal tissue and small airway wall thickness correlated strongly with the prominence of the bronchial and bronchointerstitial pulmonary patterns observed radiographically. Small airway thickness was also highly correlated with the perceived prominence of the interstitial pulmonary patterns observed radiographically, and morphometric estimates of parenchymal tissue and parenchymal collagen. There were also strong correlations between the volume density of parenchymal tissue, the percentage of parenchymal collagen, peribronchiolar mononuclear cell infiltrates, and bronchiolar mucosal plication estimates. In horses with more prominent bronchiolar mucosal plication, there was a strong direct relation to the observed prominence of peribronchiolar and submucosal blood vessels, and the bronchial and bronchointerstitial patterns observed radiographically. Horses with prominent peribronchiolar mononuclear cell infiltrates also had more obvious interstitial and bronchointerstitial pulmonary patterns observed radiographically. There also was a direct correlation between the percentage of parenchymal collagen and the observed prominence of peribronchiolar and submucosal blood vessels in these horses. In all horses, there was a strong negative correlation between the estimated average alveolar size and the observed severity of the vascular and bronchial patterns observed radiographically. Four horses with the greatest estimated airway wall and interalveolar collagen had more prominent interstitial and bronchointerstitial densities and histopathologic evidence of bronchiolitis. These horses had evidence of epithelial basement membrane disruption, with disorganized collagen fibers running between the adventitial layer and the epithelial basement membrane. Amounts of collagen were greater in the adventitia and interalveolar septa, with the fibers appearing larger and more coarse and disorganized. In horses with the greatest percentage of interalveolar septal collagen, accumulations of collagen were larger in the interalveolar septal tips. These findings suggest that horses with prominent interstitial and bronchointerstitial pulmonary patterns radiographically have undergone previous episodes of pulmonary injury, which has resulted in deposition of increased amounts of collagen in interalveolar septa and airway walls.

A histochemical and morphometric study of fiber types in a variety of skeletal muscles of healthy young adult cats was undertaken to provide normative data not available previously. Using a standardized system of nomenclature, fiber types 1, 2A, 2B, and 2C were identified in most cat muscles on the basis of myosin ATPase staining at pH 4.45. Type-2M fibers were present in temporalis (TEM) and masseter (MAS) muscles. Type-1 fibers predominated in medial head of triceps (MHT) and soleus muscles. Type-2B fibers were dominant in biceps femoris, lateral head of gastrocnemius, cranial tibial, long head of triceps, and superficial digital flexor muscles; type-2A fibers were dominant in buccinator muscle samples; and type-2M fibers were dominant in TEM and MAS muscles. Numbers of type-2C fibers did not exceed 2 to 3% of the myofiber population in any muscle. In CT and LHT muscles, a gradient of fiber type distribution was observed, with significant (P < 0.05) increase in numbers of type-1 and type-2A fibers in deeper regions of the muscles. The distribution of fiber types was compartmentalized in MHT and MAS specimens. Diameter of type-2B fibers was significantly (P < 0.05) larger than that of type-1 and type-2A fibers in biceps femoris, lateral head of gastrocnemius, cranial tibial, long head of triceps, and superficial MHT muscles. Diameter of type-2M fibers was significantly (P < 0.05) larger than that of type-1 fibers in TEM and MAS muscles. The soleus type-1 muscle fibers were the largest fibers encountered in any muscle. In MHT muscle, fiber diameter of type-1 and type-2B fibers varied significantly (P < 0.05) in oxidative and glycolytic compartments. Variability coefficients were less than 200 in all muscles. In every muscle specimen, the number of fibers with internal nuclei was less than or equal to 2%.

Bovine immunodeficiency virus (BIV), a lentivirus, is prevalent in dairy and beef cattle in southeastern United States and may be associated with a lymphoproliferative disease. The mode(s) of BIV transmission are undefined. Because artificial insemination is a common practice in dairy production, contaminated stud semen could serve as an important source of infection if the virus is harbored in seminal fluids. To evaluate this possibility, we procured 11 cryopreserved semen specimens from a stud semen repository. Leukocytes were purified from the specimens, and the leukocyte DNA was used as template in a polymerase chain reaction procedure that targeted a 235-base pair, highly conserved domain of the BIV pol gene. The target sequence was amplified from the seminal leukocyte DNA of 9 of the specimens (82%), and nucleotide sequencing confirmed the BIV specificity of the fragment. This finding provides evidence that stud bull semen may serve as an important reservoir of BIV, suggesting the possibility that artificial insemination of dairy cows may have a major role in transmission and wide-spread dissemination of this bovine lentivirus.

To measure the concentration of serum amyloid A (SAA) protein in horses a sensitive and highly reproducible sandwich (ELISA) was established, using affinity purified SAA antibody. Results of the ELISA were found to have a high correlation (r = 0.95) with those of the single radial immunodiffusion test. Equine SAA concentration was measured by use of this ELISA. In clinically normal horses, the concentration of SAA was high immediately after birth to 2 weeks of age. After that, SAA concentration had periodic fluctuations in the range of approximately 10 to 30 microgram/ml. Mean (+/- SD) concentrations of SAA in foals (less than or equal to 12 months old) and adult horses (greater than or equal to 18 months old) were 21.23 +/- 12.20 and 14.93 +/- 9.07 microgram/ml, respectively. In mares during the perinatal period, SAA concentration remained stable within the reference range before parturition. It increased quickly after delivery, and reached a peak value of 101.29 +/- 98.82 microgram/ml on postpartum day 3, then began to decrease, at postpartum week 2, to the reference range by the end of postpartum month 1. In horses with experimentally induced inflammation, SAA concentration increased quickly and reached approximately four- to 40-fold increase over the pretreatment value on day 1 and remained high on days 2 to h after treatment. It then returned to the baseline value by 2 to 4 weeks in association with disappearance of local signs of inflammation. The SAA concentration was high in most horses with clinical signs of inflammation. It was concluded from these data that this ELISA is sensitive and reliable for measuring SAA in horses.

The effects of 3 days of glucocorticoid administration on bovine blood neutrophil expression of L-selectin and CD18, and on the health status of mammary glands subclinically infected with Staphylococcus aureus were measured in 9 lactating Holsteins. The experiment was a 3 x 3 Latin square cross-over design, with 3 glucocorticoid treatments switched among groups of 3 cows/treatment during 3 periods. Treatments consisted of a vehicle (control, 10 ml of excipient/cow/d), cortisol (7.5, 15, and 7.5 mg/cow on days 1, 2, and 3, respectively), and dexamethasone (0.04 mg/kg of body weight/cow/d for total daily dosages that ranged from 21.6 to 33.2 mg). Blood samples for immunostaining and flow cytometric analysis of L-selectin and CD18 and leukograms, as well as foremilk samples for determination of S aureus shedding somatic cell counts, protein and fat percentages, and daily milk yields were collected repeatedly before, during and after treatment days. Dexamethasone caused a profound, acute, short-lived down-regulation of L-selectin on neutrophils, which correlated in time to leukocytosis, mature and immature neutrophilias, increased shedding of S aureus in infected glands, and onset of high percentages of fat and protein and decreased milk yields. Dexamethasone also caused profound but delayed down-regulation of neutrophil CD18, which reached nadir simultaneously with reappearance of L-selectin-bearing neutrophils, normalized blood neutrophil counts, markedly high foremilk somatic cell counts and protein percentage, decreased S aureus shedding in milk, and finally, expression of clinical mastitis in some infected quarters. Each of these variables had returned to control (vehicle) values by the ninth (and last) sample collection day. Although cortisol treatment also decreased expression of L-selectin and CD18 on neutrophils, dosages used in this study were not sufficient to alter the number of circulating cells or to convert subclinical mammary gland infections to clinical mastitis. These results suggest that mammary gland health status can be altered by sudden exposure of blood neutrophils to glucocorticoids, because these steroid hormones caused profound down-regulation of the adhesion molecules that direct neutrophil margination and migration through the vascular endothelium. The results also reinforce the potential disease risk of treating infected animals with potent synthetic glucocorticoids, such as dexamethasone.