Longitudinal relationship between fecal culture, fecal quantitative PCR, and milk ELISA in Mycobacterium avium ssp. paratuberculosis-infected cows from low-prevalence dairy herds.

Mycobacterium avium ssp. paratuberculosis (MAP), the causative agent of ruminant Johne's disease, presents a particular challenge with regard to infection mitigation on dairy farms. Diagnostic testing strategies to identify and quantify MAP and associated antibodies are imperfect, and certain facets of the relationship between diagnostic tests remain to be explored. Additional repeated-measures data from known infected animals are needed to complement the body of cross-sectional research on Johne's disease-testing methods. Statistical models that accurately account for multiple diagnostic results while adjusting for the effects of individual animals and herds over time can provide a more detailed understanding of the interplay between diagnostic outcomes. Further, test results may be considered as continuous wherever possible so as to avoid the information loss associated with dichotomization. To achieve a broader understanding of the relationship between diagnostic tests, we collected a large number of repeated fecal and milk samples from 14 infected cows, in addition to bulk milk samples, from 2 low-prevalence dairy herds in the northeast United States. Predominately through the use of mixed linear modeling, we identified strong associations between milk ELISA optical density, fecal quantitative PCR, and fecal culture in individual animals while concurrently adjusting for variables that could alter these relationships. Notably, we uncovered subtleties in the predictive abilities of fecal shedding level on milk ELISA results, with animals categorized as disease progressors reaching higher ELISA optical density levels. Moreover, we observed that spikes in fecal shedding could predict subsequent high ELISA values up to 2 mo later. We also investigated the presence of MAP in individual milk samples via PCR and noted an association between poor udder hygiene and MAP positivity in milk, suggesting some level of environmental contamination. The paucity of positive milk samples and the complete absence of detectable MAP in the bulk tank throughout the study period indicate that contamination of milk with MAP may not be of chief concern in low-prevalence herds. An enhanced understanding of the interrelationships between diagnostic tests can only benefit the development of testing strategies and objectives, which in turn may lessen MAP infection prevalence in dairy herds.

Utility of 2 immunological tests for antemortem diagnosis of equine protozoal myeloencephalitis (Sarcocystis neurona Infection) in naturally occurring cases.

BACKGROUND: Antemortem diagnosis of equine protozoal myeloencephalitis (EPM) is challenging. Limited information is available regarding a commercial test (surface antigen 1 [SAG-1] ELISA). Performance of another commercial test (indirect fluorescent antibody test [IFAT]) using samples from an independent group has not been well described. HYPOTHESIS/OBJECTIVES: The primary goal was to evaluate the SAG-1 ELISA and IFAT using naturally occurring EPM cases. A secondary goal was to obtain more information regarding clinical presentation. ANIMALS: Hospital cases were admitted over 20 months and classified into 4 groups. Confirmed positive cases (n = 9) had asymmetric or multifocal neurologic deficits or both and postmortem lesions consistent with EPM. Confirmed negative cases (n = 17) had variable clinical signs and postmortem lesions consistent with another neurologic disease (or no lesions). Suspected positive cases (n = 10) had asymmetric or multifocal deficits or both, marked improvement after treatment for EPM, and other likely diseases excluded. Suspected negative cases (n = 29) had orthopedic disease and no neurologic deficits. METHODS: Results of immunological testing (SAG-1 ELISA and IFAT on serum or cerebrospinal fluid [CSF] or both), neurologic examinations, CSF analyses, and postmortem examinations were analyzed retrospectively. RESULTS: SAG-1 ELISA sensitivity was 12.5% (95% CI, 1.6-38.4) and specificity was 97.1% (95% CI, 84.7-99.9) using serum. IFAT sensitivity was 94.4% (95% CI, 72.7-99.9) and specificity was 85.2% (95% CI, 66.3-95.8) using serum; sensitivity was 92.3% (95% CI, 64.0-99.8) and specificity was 89.7% (95% CI, 72.7-97.8) using CSF. CONCLUSIONS AND CLINICAL IMPORTANCE: Low sensitivity of the SAG-1 ELISA limited its usefulness for antemortem diagnosis of EPM in this patient population.

Coagulation abnormalities and complications after percutaneous liver biopsy in horses.

BACKGROUND: Liver biopsy is useful in the diagnosis of liver disease in horses. However, bleeding is the major complication. Liver dysfunction can result in abnormalities in coagulation, although there is no definitive association between risk of hemorrhage after biopsy and coagulopathy in humans. Frequency of coagulopathies in horses with liver disease and the nature and frequency of complications after liver biopsy are not reported. HYPOTHESIS/OBJECTIVES: To determine whether there is an association between coagulopathy and hemorrhage after liver biopsy in horses. ANIMALS: Horses with suspected liver disease from which a liver biopsy had been obtained. METHODS: Retrospective study. Variables indicative of coagulation were recorded. The frequency and nature of complications after biopsy were assessed using clinical and hematologic data. The association between abnormal coagulation variables and complications was assessed. RESULTS: Seventy biopsies were obtained from 66 horses. At least 1 coagulation profile abnormality was identified in 58% of the 43 horses with histopathologically confirmed liver disease. Complications were observed in 4/32 monitored horses (33 biopsies). Three horses had a decrease in the packed cell volume suggestive of subclinical bleeding, and 1 horse developed a diaphragmatic hematoma. There was no association between coagulation profile abnormality and complications. CONCLUSION AND CLINICAL IMPORTANCE: Abnormalities of coagulation are common in horses with liver disease. Liver biopsy appears to be a safe procedure in the horse. An abnormal coagulation profile is not clearly associated with an increased risk of complications after biopsy.

Prediction of Packed Cell Volume after Whole Blood Transfusion in Small Ruminants and South American Camelids: 80 Cases (2006-2016).

BACKGROUND: Calculation of desired whole blood transfusion volume relies on an estimate of an animal's circulating blood volume, generally accepted to be 0.08 L/kg or 8% of the animal's body weight in kilograms. OBJECTIVE: To use packed cell volume before and after whole blood transfusion to evaluate the accuracy of a commonly used equation to predict packed cell volume after transfusion in small ruminants and South American camelids; to determine the nature and frequency of adverse transfusion reactions in small ruminants and camelids after whole blood transfusion. ANIMALS: Fifty-eight small ruminants and 22 alpacas that received whole blood transfusions for anemia. METHODS: Retrospective case series; medical record review for small ruminants and camelids that received whole blood transfusions during hospitalization. RESULTS: Mean volume of distribution of blood as a fraction of body weight in sheep (0.075 L/kg, 7.5% BW) and goats (0.076 L/kg, 7.6% BW) differed significantly (P < 0.01) from alpacas (0.103 L/kg, 10.3% BW). Mild transfusion reactions were noted in 16% of transfusions. CONCLUSIONS AND CLINICAL RELEVANCE: The generally accepted value of 8% for circulating blood volume (volume of distribution of blood) is adequate for calculation of transfusion volumes; however, use of the species-specific circulating blood volume can improve calculation of transfusion volume to predict and achieve desired packed cell volume. The incidence of transfusion reactions in small ruminants and camelids is low.

Tissue predilection sites and effect of dose on Mycobacterium avium subs. paratuberculosis organism recovery in a short-term bovine experimental oral infection model.

The objective of this study was to develop a short-term experimental infection model for Mycobacterium avium subsp. paratuberculosis (MAP) in cattle, using small oral doses of organisms. Specifically, the effect of dose size was evaluated, as well as specific tissue predilection sites for recovery of MAP. Oral doses as low as 1.5 x 10(6) CFU reliably produced infection that could be detected 3 weeks following infection. Detection of infection required culture of multiple intestinal samples (jejunum and ileum) for MAP. Histological examination did not permit detection at this early stage. Results from this study suggest intestinal mucosa, rather than tonsil, as the primary portal of entry for MAP. The experimental infection model described here is useful for studying the early effects of preventive and therapeutic interventions for paratuberculosis in cattle.

Plasma Peak and Trough Gentamicin Concentrations in Hospitalized Horses Receiving Intravenously Administered Gentamicin.

BACKGROUND: Gentamicin is an aminoglycoside antimicrobial commonly used in horses at 6.6 mg/kg IV once daily. Therapeutic drug monitoring (TDM) can confirm desired peak concentration is reached for common bacterial isolates, and detect toxicosis associated with high trough values. OBJECTIVES: Determine the relationship between gentamicin dose and plasma concentration in hospitalized horses, and identify a starting dose range to achieve peaks > 32 µg/mL. ANIMALS: Sixty-five horses (2002-2010) receiving once-daily gentamicin with TDM performed (N = 99 sets). METHODS: Retrospective study. Data from hospitalized horses including weight, dose, plasma peak, and trough gentamicin concentration, creatinine concentrations and presence of focal or systemic disease were collected from medical records. Peak concentrations measured 25-35 minutes after administration were included (N = 77). Data were divided into low (<7.7 mg/kg), medium (7.7-9.7 mg/kg) and high (>9.7 mg/kg) dose groups, and were grouped by the horse having focal or systemic disease. RESULTS: Peak concentrations resulting from doses ≥7.7 mg/kg were 5.74 µg/mL (SE 2.1 µg/mL) greater than peaks from doses <7.7 mg/kg (P = .007). Peak concentrations was 3.6 times more likely to be >32 µg/mL if dose was ≥7.7 mg/kg (P = .04). There were no significant effects of dose on trough or creatinine concentration. At a given dose, horses with focal disease had higher peaks than those with systemic disease (P = .039). CONCLUSIONS AND CLINICAL IMPORTANCE: These data suggest gentamicin dosage should be individually determined in horses using TDM, but support an initial once-daily dose of 7.7-9.7 mg/kg IV to achieve peaks >32 µg/mL and trough concentrations <2 µg/mL. Further studies evaluating the safety of doses >6.6 mg/kg are required.

Rhodococcus equi pneumonia in 48 foals: response to antimicrobial therapy.

Case records of 48 foals with pneumonia due to Rhodococcus equi were reviewed. Twenty of the 48 foals survived and 28 died or were euthanized. There was no significant difference between the survivors and non-survivors in the age of onset of illness, duration of illness prior to admission, the mean white blood cell (WBC) count, or the mean plasma fibrinogen content. All foals had R. equi isolated from a tracheobronchial aspirate or lung specimens obtained at necropsy. All organisms were susceptible in vitro (Kirby-Bauer) to erythromycin and gentamicin. Susceptibilities to other drugs were: trimethoprim-sulfamethoxazole (88%), tetracycline (87%), chloramphenicol (83%); 97% were resistant to cephalothin and 83% to penicillin. Thirteen of the 20 surviving foals were treated with erythromycin and/or rifampin. A decline in mortality rate was observed with the introduction of the combination of erythromycin and rifampin. None of the 17 foals treated with penicillin and gentamicin survived. Chronic, active, non-septic synovitis was confirmed in 17 foals. These foals had joint distension with mild or no apparent lameness.

ELISA and fecal culture for paratuberculosis (Johne's disease): sensitivity and specificity of each method.

The sensitivity and specificity of the ELISA and fecal culture tests for paratuberculosis in dairy cattle are examined. ELISA and fecal culture data from seven dairy herds where both fecal cultures and ELISA testing was done concurrently are included. A cohort of 954 cattle including 697 parturient adults, cultured every 6 months from 10 herds followed over 4 years served as the basis to determine fecal culture sensitivity. The fecal culture technique utilized a 2g sample with centrifugation and double incubation. Of the 954 cattle cohort of all ages (calf to adult) that were fecal sampled on the first herd visit, 79 were culture positive. An additional 131 animals were detected as culture positive over the next seven tests at 6-month intervals. The sensitivity of fecal culture to detect infected cattle on the first sampling was 38%. Of the 697 parturient cattle cohort, 67 were positive on the first fecal culture, while an additional 91 adult cattle were culture positive over the next seven tests, resulting in a sensitivity of 42% on the first culture of the total animals identified as culture positive. Animals culled from the herds prior to being detected as infected and animals always fecal culture negative with culture positive tissues at slaughter are not included in the calculations. Both groups of infected cattle will lower the apparent sensitivity of fecal culture. Infected dairy herds tested concurrently with both fecal culture and ELISA usually resulted in more than twofold positive animals by culture compared to ELISA. The classification of infected cattle by the extent of shedding of Mycobacterium paratuberculosis in the feces helps define the relative proportion of cattle in each group and therefore the likelihood of detection by the ELISA test. ELISA has a higher sensitivity in animals with a heavier bacterial load, i.e. high shedders (75%) compared to low shedders (15%). Repeated testing of infected herds identifies a higher proportion of low shedders which are more likely to be ELISA negative. Thus, the sensitivity of the ELISA test decreases with repeated herd testing over time, since heavy shedders will be culled first from the herds.

Evaluation of a commercial enzyme-linked immunosorbent assay for the diagnosis of paratuberculosis in dairy cattle.

The performance of a commercially available ELISA for detection of antibodies to Mycobacterium paratuberculosis was evaluated using sera from 1,146 cows. Samples were from uninfected cattle, infected subclinical cattle shedding low numbers of organism in feces, subclinical heavy shedders, clinical cases, and randomly selected cattle in a slaughterhouse survey for paratuberculosis. The overall sensitivity of the test, using the manufacturer's recommended cutoff was 45% +/- 4.8%, and the specificity was 99% +/- 0.9%. The ELISA result was significantly correlated with the number of colonies of M. paratuberculosis detected by fecal culturing. The sensitivity of the test was highest for clinical cases of paratuberculosis (87% +/- 8.4%), and lowest for subclinical, light-shedding cattle (15% +/- 6.6%). Changing the cutoff point did not improve performance of the test. Evaluating ELISA results with a kinetic-based method reduced plate-to-plate variation in results but did not improve performance of the test based on receiver-operating characteristic curve analysis.

Variations in urinary gamma glutamyl transferase/urinary creatinine ratio in horses with or without pleuropneumonia treated with gentamicin.

The urinary GGT/urinary creatinine (uGGT/uCR) ratio was measured on Days 1, 3 and 10 in 4 adult, healthy horses; in 6 adult, healthy horses treated with gentamicin at recommended dosages and 9 adult horses treated for pleuropneumonia with gentamicin at recommended dosages. Plasma creatinine and gentamicin trough concentrations were measured on the same days. The uGGT/uCr ratio was higher in the normal horses (mean +/- s.d. 22.85 +/- 13.69) than previously reported normal values (10.5 +/- 6.8) (Adams and McClure 1985). Analysis of variance for repeated measures was used to compare the ratio in the 3 groups while controlling for the effect of time. Sick horses had a significantly higher uGGT/uCr ratio than either of the 2 groups of normal horses. Both groups of horses that were treated with gentamicin had similar percentage increases in uGGT/uCr ratio over the treatment period with the most marked increases found between treatment Days 1 and 3. The increase in uGGT/Cr ratio was predominantly a result of an increase in uGGT activity rather than a decrease in uCr concentration. The increase in uGGT activity and uGGT/uCr ratio occurred without abnormalities in serum creatinine or gentamicin trough concentrations. These findings demonstrate that urine GGT activity and uGGT/uCr ratio should be expected to increase in response to gentamicin therapy at recommended dosages without measurable changes in serum creatinine. This suggests that an elevation of the uGGT/uCr ratio in horses being treated with gentamicin would not necessarily require changes in, or withdrawal of, the gentamicin treatment as long as increases in the plasma creatinine do not exceed 0.3 g/l and gentamicin trough concentrations are < 2 micrograms/l.

Tolerance of a rice-based oral rehydration solution given to normal calves.

The objective of the study was to test the tolerance of a rice-based oral rehydration formula when fed to calves. Six healthy Holstein calves, 1 week of age, were fed the formula instead of milk replacer for 3 days. Pre- and posttreatment results of clinical examination and laboratory parameters were compared. Vital signs, attitude, appetite, clinical hydration status, urine specific gravity, and most routine serum biochemistry test results did not vary and remained within the normal range. Five of the 6 calves developed diarrhea when fed the rice-based formula, which was accompanied by a reduction in fecal pH and presence of reducing sugars in the feces. This effect was reversed when calves were returned to the milk replacer diet at the end of the study. Diarrhea was accompanied by increased water consumption, which allowed the calves to maintain normal hydration status. These results suggest that calves are unable to properly digest the rice-derived carbohydrate, and this type of formula is not recommended for oral rehydration of calves.

An echocardiographic study of atrial fibrillation in horses: before and after conversion to sinus rhythm.

Two-dimensional and M-mode echocardiograms were recorded from 41 horses before they were successfully treated for atrial fibrillation. In addition, these examinations were performed in a subgroup of 20 horses after treatment, and the results were compared with pretreatment values. Atrial fibrillation in this group of horses was associated with a reduction of mean left ventricular fractional shortening (mean 31% +/- 5.24%), and 22 of the 41 horses were below the reference range. The remaining mean M-mode variables were within the normal reference range, although 12 horses had increased left ventricular lumen dimensions in systole, and 8 horses had decreased left ventricular ejection times. Abnormal motion of the mitral valve was present in all horses and was characterized by the absence of A peaks, which were replaced by small diastolic undulations in 55% of the horses. In horses 1 to 20, after conversion to sinus rhythm, the mean fractional shortening increased (35.34% +/- 5.4%, P = .004), but there were no significant differences in heart rate or left ventricular lumen diameters in systole or diastole. These results suggest that ventricular function may be compromised by the presence of atrial fibrillation. However, this improved after correction of the arrhythmia.

Hepatic failure in dairy cattle following mastitis or metritis.

Hepatic failure developed following mastitis or metritis in five adult cows. In all five cases, cows initially showed clinical signs compatible with endotoxemia, which resolved with appropriate therapy for the mastitis or metritis. Subsequently, signs of liver failure developed including profound anorexia, weight loss, cessation of milk production, and in one case, photosensitization. Four cows had laboratory evidence of liver disease and failure (abnormally prolonged sulfobromophthalein [BSP] clearance half-life and abnormally high serum liver enzyme activity). Hepatocellular necrosis or vacuolization was seen on histopathologic examination of liver specimens from all five cows. The hepatocellular necrosis, in some cases, was thought to be due to the direct or indirect effects of endotoxin on the liver. Three of the cows responded to symptomatic therapy. One cow failed to respond and one was not treated.

Epidemiology of equine protozoal myeloencephalitis in North America based on histologically confirmed cases. A report.

Following a workshop on equine protozoal myeloencephalitis (EPM) convened at the Veterinary Medical Forum of the American College of Veterinary Internal Medicine in 1988, this survey of EPM in North America was developed. It is based upon 364 histologically confirmed case records from California, Florida, Illinois, Kentucky, New York, Ohio, Oklahoma, Ontario, Pennsylvania, and Texas up to 1988. The highest rate of infection was found in young Thoroughbred, Standardbred, and quarter horses. Differences in geographic location, sex, and month (season) of infection were not discernible. This report, the first comprehensive survey of EPM in North America, is intended to serve as a basis for evaluating future changes in prevalence and spread of EPM.

Pharmacokinetics of intravenously and intramuscularly administered ticarcillin and clavulanic acid in foals.

Serum concentrations of ticarcillin and clavulanic acid were measured in healthy foals (2 to 6 months old) given the drugs in combination by intravenous and intramuscular routes of administration. Five foals were administered 50 mg of ticarcillin/kg of body weight and 1.67 mg of clavulanic acid/kg, IV. Five foals were administered 100 mg of ticarcillin/kg and 3.33 mg of clavulanic acid/kg, IV, and 4 of those 5 were given the same combined dose IM. The elimination half-life of ticarcillin for intravenous administration was 0.83 hour for the low dosage and 0.96 hour for the high dosage. After intramuscular administration, the half-life of elimination was 2.9 hours, with bioavailability of 54.6%. For IV administered clavulanic acid, the elimination half-life was 0.65 hour for the low dosage and 0.74 hour for the high dosage. After intramuscular administration, the elimination half-life was 0.92 hour, and bioavailability was 68.1%. A combined dosage, 50 mg of ticarcillin/kg and 1.67 mg of clavulanic acid/kg, given every 6 hours is recommended.

Pharmacokinetics of enrofloxacin in adult horses and concentration of the drug in serum, body fluids, and endometrial tissues after repeated intragastrically administered doses.

OBJECTIVE: To investigate the pharmacokinetics of enrofloxacin in adult horses. DESIGN: 2-dose oral and i.v. cross-over trial followed by multiple oral doses. ANIMALS: 8 clinically normal adult horses. PROCEDURE: Enrofloxacin was administered at dosages of 2.5 mg/kg of body weight to 4 horses and 5.0 mg/kg to 4 other horses. Each dose was given by the intragastric and i.v. routes, using a cross-over design. After the first intragastric dose, 5 additional doses were administered at 12-hour intervals. Enrofloxacin concentrations were measured in serum, synovial fluid, peritoneal fluid, urine, CSF, and endometrial tissues. RESULTS: Disposition of enrofloxacin after i.v. administration conformed to a 2-compartment model with an elimination half-life of 5.95 and 6.09 hours for the low and high dose, respectively. After the first intragastric administration, time to peak concentration was 1.0 +/- 0.35 and 1.25 +/- 0.43 hours, and the bioavailability was 57.39 +/- 8/45 and 62.52 +/- 19.65% for the low and high dose, respectively. After multiple intragastric administration, peak serum concentration (at 72 to 96 hours) was 2.62 +/- 0.61 micrograms/ml for the low dose and 5.97 +/- 1.56 micrograms/ml for the high dose. After multiple intragastric doses, synovial fluid, urine, and endometrial tissue concentrations exceeded serum concentrations. Peritoneal fluid and CSF concentrations were lower than serum concentrations. CONCLUSIONS: Computer modeling of the pharmacokinetic variables suggested that a single daily i.v. administered dose of 5.5 mg/kg, or orally administered doses of 7.5 mg/kg every 24 hours or 4.0 mg/kg every 12 hours, would be effective in horses. Additional studies are necessary to confirm the efficacy and safety of these dosages in a clinical setting.

Pharmacokinetics of cefotaxime in neonatal pony foals.

Serum concentrations of cefotaxime and desacetylcefotaxime were measured in 1-week-old pony foals after IV administration of a single dose of cefotaxime. The cefotaxime disposition data conformed to a two-compartment model with elimination half-life of 0.60 hour. The combined cefotaxime and desacetylcefotaxime data was best described by a four-compartment model. The apparent half-life describing the disappearance of desacetylcefotaxime was 1.69 hours. Dosage of 40 mg/kg of body weight given IV every 4 to 6 hours for neonatal foals with gram-negative septicemia and every 2 hours for foals with meningitis is recommended for further study.

Disposition and excretion of flunixin meglumine in horses.

The disposition of flunixin meglumine administered IV at a dosage of 1.1 mg/kg was described by a 2-compartment model; the alpha and beta half-lives (t1/2) were 0.61 and 1.5 hours, respectively. When administered IV at a rate of 2.2 mg/kg, the disposition was best described by a 3-compartment model, and the alpha, beta, and lambda t1/2 were 0.16, 1.52, and 6.00 hours, respectively. The zero-time plasma concentrations after flunixin meglumine was administered at 1.1 and 2.2 mg/kg were 9.3 +/- 0.76 and 21.5 +/- 7.4 mg/L, respectively. The bioavailability after oral administration of 1.1 mg/kg was 85.8%. The absorption t1/2 was 0.57 hours, with a peak concentration of 2.50 +/- 1.25 mg/L. The cumulative urinary recoveries for IV and oral administrations were 61.0% and 63.3%, respectively, of the dose for the 12-hour collection period. The final asymptotic points of urine excretion after IV and oral administrations were 406.4 +/- 65.5 and 357.7 +/- 53.5 mg, respectively, which represented 75.5 and 77.5% of the drug accounted for between 30 and 35 hours after administration. Flunixin meglumine was rapidly excreted in urine over a 2- to 4-hour period after drug administration and was highly bound to protein in plasma.