Objective-To determine the prevalence of biofilm formation under long-term cell culture conditions in serum samples of dairy cattle, goats, cats, and dogs, and to determine whether there is an association between nanobacteria and biofilm formation. Sample Population-Serum samples of clinically normal animals (313 dairy cattle, 48 goats, 140 dogs, and 44 cats) and animals with various medical conditions (60 dogs and 116 cats). Procedure-Serum was incubated under cell culture conditions and observed for biofilm formation by use of light microscopy, electron microscopy, and spectroscopy. A polymerase chain reaction assay was developed to identify 16S rRNA gene sequences of nanobacteria. Results-Biofilm formation developed in serum samples of 304 of 313 (97%) cattle, 44 of 48 (92%) goats, 44 of 44 (100%) cats, and 126 of 140 (90%) dogs. Prevalence of serum samples with positive results for biofilm formation was not significantly different between cats or dogs with and without medical conditions associated with pathologic extraskeletal calcification processes. Scanning electron microscopy and spectroscopy of biofilm samples revealed small coccoid particles consisting mainly of calcium and phosphate. Polymerase chain reaction assay failed to amplify sequences of nanobacteria. Conclusions and Clinical Relevance-Under longterm cell culture conditions, biofilm made up of aggregates of calcium and phosphate crystals does form in serum samples of clinically normal dairy cattle, goats, cats, and dogs. Disease, however, does not predispose to biofilm formation in serum samples of dogs and cats. Our findings did not support the existence of nanobacteria in serum samples of cattle, goats, cats, and dogs.

Objective-To evaluate effects of sedation on stability of resistance of the respiratory system (RRS) and measures of resting energy expenditure (REE) by use of open-flow indirect calorimetry (IC) and treatment with aerosolized albuterol on REE in horses with recurrent airway obstruction (RAO). Animals-9 clinically normal horses and 8 horses with RAO. Procedure-In phase 1, RRS was measured by using forced oscillometry (FOT) in 5 clinically normal horses before and after sedation with xylazine. In phase 2, REE was measured in 4 clinically normal horses between 20 and 25 minutes and again 35 to 40 minutes after sedation with xylazine. In phase 3, IC was performed between 20 and 25 minutes and FOT was performed between 30 and 35 minutes after xylazine administration in 8 horses with RAO; after administration of 450 µg of albuterol, IC and FOT were repeated. Results-In phase 1, RRS values were significantly lower 5 and 10 minutes after sedation. In phase 2, diminishing sedation did not significantly affect REE. In phase 3, there was a significant decrease in mean RRS (1.15 +/- 0.25 vs 0.84 +/- 0.14 cm H20/L/s) and REE (30.68 +/- 17.89 vs 27.46 +/- 16.54 kcal/kg/d) after albuterol administration. Conclusions and Clinical Relevance-FOT and IC are useful in obtaining repeatable measurements of RRS and REE, respectively, in sedated horses. Concurrent bronchodilation and decreased REE after albuterol administration suggest that increased work of breathing as a result of airway obstruction may contribute to increased energy demands in horses with RAO.

Objective-To determine whether pharmacokinetic analysis of data derived from a single IV dose of iohexol could be used to predict creatinine clearance and evaluate simplified methods for predicting serum clearance of iohexol with data derived from 2 or 3 blood samples in clinically normal foals. Animals-10 healthy foals. Procedure-Serum disposition of iohexol and exogenous creatinine clearance was determined simultaneously in each foal (5 males and 5 females). A 3-compartment model of iohexol serum disposition was selected via standard methods. Iohexol clearance calculated from the model was compared with creatinine clearance. Separate limited-sample models were created with various combinations of sample times from the terminal slope of the plasma versus time profile for iohexol. Correction factors were determined for the limited-sample models, and iohexol clearance calculated via each method was compared with exogenous creatinine clearance by use of method comparison techniques. Results-Mean exogenous creatinine clearance was 2.17 mL/min/kg. The disposition of iohexol was best described by a 3-compartment open model. Mean clearance value for iohexol was 2.15 mL/min/kg and was not significantly different from mean creatinine clearance. A method for predicting serum iohexol clearance based on a 2-sample protocol (3- and 4-hour samples) was developed. Conclusions and Clinical Relevance-Iohexol clearance can be used to predict exogenous creatinine clearance and can be determined from 2 blood samples taken after IV injection of iohexol. Appropriate correction factors for adult horses and horses with abnormal glomerular filtration rate need to be determined.

Objective-To compare isolates of Rhodococcus equi on the basis of geographic source and virulence status by use of pulsed-field gel electrophoresis (PFGE). Sample Population-290 isolates of R equi(218 virulent isolates from foals and 72 avirulent isolates from feces, soil, and respiratory tract samples) obtained between 1985 and 2000 from horses and horse farms from 4 countries. Procedure-DNA from isolates was digested with the restriction enzyme AseI and tested by use of PFGE. Products were analyzed for similarities in banding patterns by use of dendrograms. A similarity matrix was constructed for isolates, and the matrix was tested for nonrandom distributions of similarity values with respect to groupings of interest. Results-There was little grouping of isolates on the basis of country, virulence status, or region within Texas. Isolates of R equi were generally < 80% similar, as determined by use of PFGE. Isolates from the same farm generally were rarely of the same strain. Conclusions and Clinical Relevance-Considerable chromosomal variability exists among isolates of R equi obtained from the same farm, sites within Texas, or among countries from various continents. Only rarely will it be possible to link infections to a given site or region on the basis of analysis of isolates by use of PFGE of chromosomal DNA.

Objective-To explore whether early analysis of spatial data may result in identification of variables associated with epidemic spread of foot and mouth disease. Sample Population-37 farms with infected cattle (ie, case farms) reported within the first 6 days of the 2001 Uruguayan foot-and-mouth disease epidemic. Procedure-A georeferenced database was created and retrospective analysis was performed on case farm location in relation to farm density, cattle density, farm type (ie, beef vs dairy cattle production), road density, case farm distance to the nearest road, farm size, farm ownership, and day of infection. Mean or median results of 1 to 3 day versus 4 to 6 day spatial data were compared. Spatial-temporal associations were investigated by correlation analysis. Results-Comparison of mean or median values between the first 3 days and days 4 to 6 of the epidemic and results of correlation analysis indicated a significant increase in road density, cattle density, and dairy cattle production and a significant decrease in farm size and case farm distance to the nearest road that developed over time. A route that linked most case farms by the shortest possible distance and also considered significantly associated variables was created. It included 86.1% of all case farms reported by 60 days into the epidemic. Conclusions and Clinical Relevance-Epidemic direction can be assessed on the basis of road density and other spatial variables as early as 6 days into an epidemic. Epidemic control areas may be more effectively identified if local and regional georeferenced data are considered.

Objective-To evaluate response of euthyroid cats to administration of recombinant human thyroid-stimulating hormone (rhTSH). Animals-7 healthy cats. Procedure-Each cat received each of 5 doses of rhTSH (0, 0.025, 0.050, 0.100, and 0.200 mg), IV, at 1-week intervals. Serum concentration of total thyroxine (TT4) and free thyroxine (fT4) was measured immediately before each injection (time 0) and 2, 4, 6, and 8 hours after administration of each dose. Results-Overall TT4 response did not differ significantly among cats when administered doses were ≥ 0.025 mg. Serum TT4 concentrations peaked 6 to 8 hours after administration for all doses greater than 0.025 mg. For all doses greater than 0.025 mg, mean +/- SEM TT4 concentration at 0, 6, and 8 hours was 33.9 +/- 1.7, 101.8 +/- 5.9, and 101.5 +/- 5.7 nmol/L, respectively. For all doses greater than 0.025 mg, mean fT4 concentration at 0, 6, and 8 hours was 38.7 +/- 2.9, 104.5 +/- 7.6, and 100.4 +/- 8.0 pmol/L, respectively. At 8 hours, the fT4 response to 0.025 and 0.050 mg was less than the response to 0.100 and 0.200 mg. Adverse reactions after rhTSH administration were not detected. Conclusions and Clinical Relevance-The TSH stimulation test can be performed in cats by IV administration of 0.025 to 0.200 mg of rhTSH and measurement of serum TT4 concentrations at time of injection and 6 or 8 hours later. Clinical validation of the TSH stimulation test would facilitate development of additional tests of thyroid gland function, such as a TSH assay.

The time available to implement successful control measures against epidemics was estimated. Critical response time (CRT), defined as the time interval within which the number of epidemic cases remains stationary (so that interventions implemented within CRT may be the most effective or least costly), was assessed during the early epidemic phase, when the number of cases grows linearly over time. The CRT was calculated from data of the 2001 foot-and-mouth disease (FMD) epidemic that occurred in Uruguay. Significant regional CRT differences (ranging from 1.4 to 2.7 days) were observed. The CRT may facilitate selection of control measures. For instance, a CRT equal to 3 days would support the selection of measures, such as stamping-out, implementable within 3 days, but rule out measures, such as post-outbreak vaccination, because intervention and immunity building require more than 3 days. Its use in rapidly disseminating diseases, such as FMD, may result in regionalized decision-making.

Objective-To determine magnitude and duration of the effect of oral administration of methazolamide at 2 dosages on intraocular pressure (IOP) in dogs in single- dose and multiple-dose trials and to determine aqueous humor flow rate (AHFR) by use of anterior segment fluorophotometry before and during treatment. Animals-25 healthy adult Beagles. Procedure-Baseline IOPs and AHFRs were determined on days 0 and 1, respectively. On day 2, the single-dose trial was initiated with oral administration of 25 or 50 mg of methazolamide at 7 AM to 2 groups of 10 dogs each. Five dogs served as controls. In the multiple-dose trial, the same dogs received 25 or 50 mg of methazolamide at 7 AM and at 3 and 11 PM on days 3 through 9. Results-Intraocular pressures varied diurnally with highest IOPs in the morning. In the single-dose trial, IOP decreased significantly at 3 to 6 hours after treatment and then increased significantly at later time points, compared with baseline values. In the multipledose trial, dogs in both treatment groups had significantly lower IOPs during the treatment period at 10 AM and 1 PM but not at 6 and 9 PM, compared with baseline values. In both treatment groups morning IOPs had returned to baseline values by the first day after treatment. Evening IOPs were significantly increased by 2 to 3 days after treatment, compared with baseline values. The AHFRs in both treatment groups were significantly lower than pretreatment AHFRs. Conclusions and Clinical Relevance-Oral administration of methazolamide decreases IOPs and AHFRs in clinically normal dogs, with effectiveness diminishing in the evening.

Objective-To use threshold concentrations of acetone and beta-hydroxybutyrate in milk and serum, respectively; identify risk for ketosis and endometritis; and assess analyses of blood and milk samples as predictors of risk for ketosis in high-yielding dairy cows. Animals-90 multiparous Holstein cows. Procedure-At intervals before and after parturition, blood samples were obtained for determination of glucose, nonesterified fatty acids, leptin, insulin, insulin-like growth factor-1, and beta-hydroxybutyrate concentrations. Samples of milk were obtained at similar intervals after parturition for determination of fat content and concentrations of acetone, protein, and lactose. Reproductive examination of each cow was performed weekly. Results-For each cow, threshold concentrations of acetone and β-hydroxybutyrate were calculated as 75th and 90th percentiles of maximum postpartum concentrations of acetone in milk (0.40 and 0.87 mmol/L) and β-hydroxybutyrate in serum (2.30 and 3.51 mmol/L). Significant decrease in milk production (442 to 654 kg of energy-corrected milk/305-day period per cow) was associated with acetone or beta-hydroxybutyrate in excess of threshold values. Milk acetone concentrations > 0.40 mmol/L were associated with 3.2 times higher risk for endometritis. Low plasma glucose, high serum beta-hydroxybutyrate, and high milk acetone concentrations during week 1 after parturition were indicators of increased risk for ketosis later during lactation. Conclusions and Clinical Relevance-Determination of milk acetone concentration during the week after parturition may identify cows at risk for ketosis and endometritis; with appropriate interventions, development of disease and production losses may be reduced.

Objective-To determine whether the nonsteroidal anti-inflammatory drugs (NSAIDs) carprofen, flunixin meglumine, and phenylbutazone have cyclooxygenase (COX)-independent effects that specifically inhibit activation of the proinflammatory transcription factor nuclear factor kappa B (NfκB). Study Population-Purified ovine COX-1 and -2 and cultures of RAW 264.7 murine macrophages. Procedure-The COX-1 and -2 inhibitory effects of the NSAIDs were tested in assays that used purified ovine COX-1 and -2. Prostaglandin production was analyzed by use of a radioimmunoassay. Inhibitory effects of these drugs on lipopolysaccharide (LPS)- induction of inducible nitric oxide synthase (iNOS) and LPS-stimulated translocation of NfκB were determined by use of RAW 264.7 murine macrophages. Results-Flunixin meglumine and phenylbutazone were selective inhibitors of COX-1. Carprofen and flunixin meglumine, but not phenylbutazone, inhibited LPS-induction of iNOS. Carprofen and, to a lesser degree, flunixin meglumine had inhibitory effects on NFκB activation. Conclusions and Clinical Relevance-The ability of drugs such as carprofen and flunixin meglumine to inhibit activation of NfκB-dependent genes such as iNOS, in addition to their effects on COX, suggests an additional mechanism for their anti-inflammatory effects and may explain the ability of flunixin meglumine to be an effective inhibitor of the effects of endotoxin in horses with endotoxemia.