Synovial lesions in experimental canine Lyme borreliosis.

Borrelia burgdorferi is the causative agent of Lyme disease, which is mainly characterized by lameness in dogs. More than 95% of naturally infected dogs are asymptomatic or subclinical; however, in experimental studies, histologic synovial lesions are consistently observed in asymptomatic dogs inoculated with B. burdgorferi. This study investigates the ability of a synovial histopathologic scoring system, clinicopathologic data, and polymerase chain reaction (PCR) testing to differentiate between B. burgdorferi-infected and uninfected dogs. Eighteen 18-week-old beagles were subject to challenge with B. burgdorferi-infected wild-caught ticks (Ixodes scapularis), and 4 uninfected dogs served as controls. Infection was confirmed by serology (ELISA) and PCR amplification of B. burgdorferi-specific DNA of skin biopsies taken at the tick attachment site. A synovial scoring system from human medicine was adapted and implemented on postmortem synovial samples to discriminate infected and noninfected animals. Application of this system to elbows and stifles with a cumulative joint score cutoff  > 4 showed a sensitivity of 88.2% and a specificity of 100%, with a positive likelihood ratio of infinity and a negative likelihood ratio of 0.12. Complete blood count, serum biochemistry, urinalysis, urine protein:creatinine, urine PCR, synovial and lymph node cytology, and synovial PCR were evaluated but were not reliable indicators of clinical disease.

The anamnestic serologic response to vaccination with a canarypox virus-vectored recombinant West Nile virus (WNV) vaccine in horses previously vaccinated with an inactivated WNV vaccine.

A new recombinant West Nile virus (WNV) vaccine has been licensed for use in horses. Prior to the availability of the recombinant vaccine in 2004, the only equine WNV vaccine available on the market had been an inactivated vaccine. Since the recombinant vaccine only expresses selected viral genes, the question could be posed as to whether a single dose of the recombinant vaccine would be effective in producing an anamnestic serologic response in horses previously vaccinated with an inactivated WNV vaccine. In this study we demonstrate that vaccination of horses with a canarypox-vectored recombinant vaccine, under field conditions, results in a marked anamnestic response in horses previously vaccinated with an inactivated WNV vaccine.

Comparison of the safety and efficacy of a recombinant feline leukemia virus (FeLV) vaccine delivered transdermally and an inactivated FeLV vaccine delivered subcutaneously.

The efficacy of a new recombinant FeLV vaccine (rFeLV), delivered transdermally via a needle-free delivery device was compared to that of an inactivated FeLV vaccine (FeLV-k), administered subcutaneously, with a conventional needle and syringe. Kittens were immunized with either rFeLV (0.25 ml, transdermal) or FeLV-k (1 ml, subcutaneous); or they were sham-vaccinated with physiologic saline (0.25 ml, transdermal). Two vaccinations were administered 21 days apart. Injection sites were monitored for any acute or subacute reactions relative to vaccine administration. Four weeks following the final vaccination, all cats were subject to oro-nasal FeLV challenge. Blood was collected for determination of FeLV antigenemia (p27) at weekly intervals beginning three weeks post-challenge. All of the vaccinated cats from both groups resisted FeLV challenge; and 90% of the control cats developed persistent FeLV antigenemia in response to challenge. No acute or persistent injection site reactions were observed. The rFeLV, delivered transdermally, provides protection against persistent FeLV antigenemia following a robust challenge that is equivalent to that of FeLV-k.

Ureteral ligation prevents the haemodynamic effect of frusemide in pentobarbitol anaesthetised horses.

Frusemide reduces pulmonary vascular pressures in resting horses and attenuates exercise-induced increases in these pressures in exercising horses. The mechanism underlying these effects of frusemide is unclear. We tested the hypothesis that the haemodynamic effects of frusemide are dependent on diuresis by examining the effect of frusemide in anaesthetised horses in which diuresis was prevented by ligation of ureters. Twenty four horses were assigned randomly to one of 4 treatments: 1) frusemide (1 mg/kg bwt i.v.) and intact ureters; 2) frusemide and ligated ureters; 3) saline placebo and ligated ureters; and 4) frusemide and phenylbutazone (4.4 mg/kg bwt i.v. 12 h and 15 min before frusemide) and ligated ureters. Frusemide administration to anaesthetised horses with intact ureters increased plasma total protein concentration and reduced mean right atrial, pulmonary artery and aortic pressures. There was no significant effect of frusemide administration on haemodynamic variables or plasma total protein concentration in horses with ligated ureters. The combination of frusemide and phenylbutazone increased mean right atrial, pulmonary artery and aortic pressures in horses with ligated ureters. This study demonstrates that, in anaesthetised horses, the haemodynamic effect of frusemide is dependent upon diuresis. We interpret these results as providing further evidence that the haemodynamic effect of frusemide in horses is attributable to a reduction in plasma and blood volume.

The digital pathologies of chronic laminitis.

This review indicates that the patient-to-patient uniqueness commonly seen in chronic laminitis represents the variable presence of the digital pathologies. Although some degree of mechanical failure is always present, the secondary metabolic and growth dysplasias, vascular pathologies, and sepsis may or may not be evident. The presence and severity of these pathologies appear to have a more significant impact on the prognosis of individual cases than does the displacement of the distal phalanx. It should be reiterated that it is often the combined presence of these individual pathologies that gives rise to the patient that is totally refractory to treatment. In the absence of these pathologies, many horses with significant displacement of the distal phalanx are not in pain and are not in need of treatment. It thus follows that a key to the improved rehabilitation of difficult patients is focusing research on the physiopathology and diagnosis of these nonmechanical problems.

The pathophysiology of chronic laminitis. Pain and anatomic pathology.

The potential pathologic manifestations of chronic laminitis are just as varied, and possibly more so, than the list of possible inciting agents of the disease itself. The extent to which rehabilitation and return to normal function can be attained, cannot always be accurately determined by physical examination. It should be remembered that significant physiologic and pathologic alterations occur in chronic laminitis; thus, even if radiographically the patient returns to a normal appearance, residual morphologic and structural defects are likely to remain.

Cardiorespiratory effects of a tiletamine/zolazepam-ketamine-detomidine combination in horses.

OBJECTIVE: To determine cardiorespiratory effects of a tiletamine/zolazepam-ketamine-detomidine (TZKD) combination in horses. ANIMALS: 8 healthy adult horses. PROCEDURE: Horses were instrumented for measurement of cardiorespiratory, acid-base, and electrolyte values. Each horse was given xylazine (0.44 mg/kg of body weight, IV) 10 to 15 minutes prior to induction of recumbency by administration of the TZKD combination. Cardiorespiratory, acid-base, and electrolyte values were measured at 5-minute intervals for > or =30 minutes. RESULTS: All horses became recumbent within 1 minute after IV administration of TZKD. Mean +/- SD duration of recumbency was 40+/-8 minutes. All horses regained standing position after < or =2 attempts. Quality of anesthesia and analgesia was determined to be satisfactory in all horses. Xylazine induced decreases in respiratory rate, heart rate, cardiac output, maximum rate of increase of right ventricular pressure, and rate pressure product. The PaCO2, right atrial pressure, and peripheral vascular resistance increased, whereas blood temperature, PO2, pHa, HCO3-, PCV, total solids, Na, and K values remained unchanged. Subsequent administration of TZKD caused right atrial pressure and PaCO2 to increase and PaO2 to decrease, compared with values obtained after xylazine administration. Remaining cardiorespiratory, acid-base, hematologic, and electrolyte values did not differ from those obtained after xylazine administration. CONCLUSION: IV administration of TZKD induces short-term anesthesia in horses. Potential advantages of this drug combination are the small volume of drug administered; minimal cardiorespiratory depression; quality of induction and maintenance of, and recovery from, anesthesia; and duration of drug effects.

The pharmacokinetics of furosemide in anaesthetized horses after bilateral ureteral ligation.

The pharmacokinetics of furosemide were investigated in anaesthetized horses with bilateral ureteral ligation (BUL) with (n = 5) or without (n = 5) premedication with phenylbutazone. Horses were administered an intravenous (i.v.) bolus dose of furosemide (1 mg/kg) approximately 60-90 min after BUL. Plasma samples collected up to 3 h after drug administration were analysed by a validated high performance liquid chromatography method. Median plasma clearance (CLp) of furosemide in anaesthetized horses with BUL was 1.4 mL/min/kg. Apparent steady state volume of distribution (Vd(ss)) ranged from 169 to 880 mL/kg and the elimination half life (t1/2) ranged from 83 min to 209 h. No differences in plasma concentration or kinetic parameter estimates were observed when phenylbutazone was administered before furosemide administration. BUL markedly reduces the elimination of furosemide in horses and models the potential effects that severe changes in kidney function may have on drug kinetics in horses.

Evaluation of a portable clinical analyzer in a veterinary hospital setting.

OBJECTIVE: Evaluation of a portable clinical analyzer for determination of blood gas tensions, electrolyte and glucose concentrations, and Hct in a hospital setting. DESIGN: Prospective study. ANIMALS: 50 dogs, 50 cats, and 28 horses, all clinically normal. PROCEDURE: Blood samples were analyzed on a portable clinical analyzer to determine concentrations of sodium, potassium, chloride, BUN, glucose, and ionized calcium and values of Hct, pH, PCO2, and PO2. Values obtained were compared with those obtained from the same blood samples, using a standard automatic analyzer (serum sodium, potassium, chloride, BUN, and glucose concentrations), a cell counter (Hct), a blood gas analyzer (pH, PCO2, PO2), and a calcium-pH analyzer (ionized calcium). Bias (mean difference between values obtained on the same sample by different methods) and variability (SD of differences) were determined for all values. Data were also subjected to Deming regression analysis. RESULTS: Correlation coefficients were > 0.90 for all values except potassium and ionized calcium concentrations. Bias and variability were within clinically acceptable limits (+/- 2 SD) for all but potassium, ionized calcium, and glucose concentrations and Hct. Species-dependent variability was observed for glucose concentration and Hct. CLINICAL IMPLICATIONS: Most differences between values obtained with the portable clinical analyzer and standard clinical laboratory systems could be accounted for by differences in type of sample tested (blood vs serum). The portable clinical analyzer is suitable for point-of-care analysis in critical care situations and for routine blood biochemical analysis when extensive laboratory support is unavailable.

Accuracy of noninvasive oxyhemoglobin saturation, end-tidal carbon dioxide concentration, and blood pressure monitoring during experimentally induced hypoxemia, hypotension, or hypertension in anesthetized dogs.

OBJECTIVE: To determine reliability of noninvasive methods of arterial oxyhemoglobin saturation (SpO2), end-tidal CO2 concentration (PEtCO2), and blood pressure (BP) determination during periods of hypoxemia and systemic arterial BP perturbations. ANIMALS: 7 healthy, conditioned dogs weighing 19 to 22 kg. PROCEDURE: 3 pulse oximeters, 2 capnometers, and 2 oscillometric BP monitors were used to measure oxygen-carrying capacity of the blood, heart rate, ventilatory status and arterial BP changes during hypoxemia, and altered arterial BP. Pulse oximeter-derived SpO2 and PEtCO2 were determined during rapidly induced plateaus of hypoxia (decreased fractional in-spired oxygen concentration [FiO2]) and altered systemic arterial BP. A lead-II ECG was used to monitor heart rate. RESULTS: Pulse oximetry provided an accurate assessment of fractional oxyhemoglobin saturation (SaO2) at SpO2 > 70%. As SaO2 decreased from 70%, the magnitude of the SpO2 error increased (20% error at SpO2 < 30%). The PEtCO2, was accurate at PaCO2, ranging from 30 to 55 +/- 5 mm of Hg under all experimental conditions. When PaCO2 was > 55 mm of Hg, both capnometers produced values that were as much as 20 mm of Hg less than the corresponding PaCO2. Mean BP was least dependent on pulse wave quality, consistently underestimating mean arterial BP by approximately 10 mm of Hg. CONCLUSIONS AND CLINICAL RELEVANCE: The pulse oximeters tested provided an accurate estimation of SaO2 at SpO2 > 70%. A PEtCO2 value > 55 mm of Hg may represent hypercapnia that is more profound than indicated. Systolic BP determinations were most accurate during hypotensive states and least accurate during hypertension. Diastolic BP measurements were generally more accurate during hypertension than normotension. Accuracy is not appreciably affected by hypotension resulting from vasodilation or blood loss. The tendency to underestimate systemic arterial BP should not interfere with trend detection during unstable clinical conditions.

Cardiorespiratory effects of sevoflurane, isoflurane, and halothane anesthesia in horses.

OBJECTIVE: To determine and compare cardiorespiratory and recovery effects of sevoflurane, isoflurane, and halothane in horses. ANIMALS: 8 clinically normal horses (4 mares, 4 geldings), 5 to 12 years old. PROCEDURE: Inhalation anesthesia was maintained for 90 minutes with sevoflurane, isoflurane, or halothane. Anesthesia depth was maintained at 1.5 minimum alveolar concentration of halothane, isoflurane, and sevoflurane, then was reduced at 30 and 60 minutes. A surgical plane of anesthesia was reinduced by administration of ketamine or thiopental or by increasing the fractional inspired concentration of sevoflurane. Cardiovascular and pulmonary variables were recorded and compared among inhalation anesthetics. Recovery was monitored, and subjective assessment of recovery quality was performed. RESULTS: Hemodynamic and pulmonary indices during sevoflurane anesthesia were similar to those of isoflurane. Cardiac output and systemic arterial pressure decreased less during sevoflurane and isoflurane anesthesia than during halothane anesthesia. After 90 minutes, cardiac output was greater for sevoflurane and isoflurane, respectively, compared with halothane. Mean arterial pressure was similar for all three anesthetic agents. Respiratory rate for sevoflurane and isoflurane was less than that for halothane. This apparent respiratory depression correlated with greater increase in PaCO2 and decreased pH when sevoflurane and isoflurane were compared with halothane. Recovery from sevoflurane anesthesia was qualitatively similar and superior to recovery from isoflurane and halothane, respectively. Time to standing did not differ significantly between sevoflurane and isoflurane, but was shorter than halothane. CONCLUSIONS: Sevoflurane induced cardiorespiratory effects that were comparable to those of isoflurane and halothane. Cardiac output was greater and respiratory rate was less than that for halothane at 1.5 MAC. Sevoflurane anesthesia was characterized by good control of anesthesia depth during induction, maintenance, and recovery. Recovery time after sevoflurane anesthesia was comparable to that for isoflurane, and recovery was smooth and controlled in a manner consistent with recovery from halothane.

Vascular perfusion in horses with chronic laminitis.

Vascular perfusion casts were used to define and characterise the macroscopic perfusion defects present in the distal digit of 11 horses affected by chronic laminitis. Five clinically normal horses were used as controls. Based on clinical history and clinical status, horses with chronic laminitis were classified as being potentially treatable or clinically refractory. Eleven macroscopic vascular defects were noted in the casts from horses with laminitis. Four types of lesions were identified in the submural laminar circulation, 3 in the coronary bed and 4 were associated with the solar circulation. Multiple defects were present and a definite trend was noted for the perfusion defects to be worse in the casts of clinically refractory subjects than in those considered treatable. This information suggests that evaluation of circulatory status should add significantly to the ability to separate treatable from clinically refractory patients. Results also indicated that ventral displacement of the third phalanx (sinkers) and compression of the solar vasculature are more prevalent than is presently thought.

Keratin and associated proteins of the equine hoof wall.

In this study, we described water-insoluble proteins extracted from the germinative regions (stratum internum and coronary band epithelium) and the cornified outer surface (stratum medium) of the equine hoof wall. Two major types of polypeptides were identified: the intermediate filaments (IF) and the IF-associated proteins. The IF, including keratins, composed a major portion of this fraction, had electrophoretic mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the range of 40 to 80 kDa, and reacted with acidic or basic keratin-specific monoclonal antibodies. Differences in the composition of keratins between germinative layers and the stratum medium were seen. Another less well-characterized group of polypeptides associated with the IF also were extracted with the water-insoluble polypeptide fraction. These associated proteins had an apparent molecular weight between 10 and 30 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and contained a higher percentage of sulfur-containing amino acids than did the IF. Water-insoluble protein fractions compared favorably with those found in other less-specialized keratinizing tissue with respect to size, immunoreactivity with monoclonal antibody, and amino acid composition.

Characterisation and distribution of epidermal growth factor receptors in equine hoof wall laminar tissue: comparison of normal horses and horses affected with chronic laminitis.

Epidermal growth factor (EGF) receptors were detected in plasma membrane preparations of equine hoof wall laminar tissue at concentrations comparable to that of equine liver. Scatchard analysis of the equilibrium binding data suggested the presence of two classes of EGF binding sites in most of the controls (plasma membranes from clinically normal horses); a high-affinity class and a more numerous low-affinity class. The dissociation constant of the low-affinity class of EGF-specific receptors (KD = 1 x 10(-9)M) is in reasonable agreement with other values established for the EGF receptor. The variability between individual estimates for the KD of the high-affinity receptor class precluded an accurate estimate for those sites. A possible explanation is discussed. The high-affinity binding sites were uniformly absent in plasma membranes prepared from horses affected by chronic laminitis. Autoradiographic analysis localised the EGF receptors primarily to the secondary epidermal laminae, with an apparent greater density over the proliferative basal keratinocytes. Little label was associated with the dermal or the keratinised primary epidermal laminae. Tissue from horses with chronic laminitis had EGF receptors located uniformly over the hyperplastic epidermal keratinocytes. These data suggest that an EGF-mediated response may be involved in the hyperproliferative response that is characteristic of chronic laminitis.

EGF receptor-binding activity in the urine of normal horses and horses affected by chronic laminitis.

A heterologous radioreceptor binding assay (RRA) has been developed capable of detecting nanogram amounts of epidermal growth factor (EGF) receptor-binding activity in equine urine. The binding parameters of [125I]mEGF (murine EGF) to EGF receptors on equine plasma membranes are in good agreement with values from other EGF-RRA systems. The dissociation constant estimated from equilibrium methods (KD = 4 X 10(-10) M) is in reasonable agreement with that determined from the rate constants (KD = 6 X 10(-10) M) and is in good agreement with values determined in other species. The assay is specific for equine EGF (eEGF) receptor-binding activity and capable of detecting less than 0.34 nM eEGF receptor-binding activity in urine. Equine EGF receptor-binding activity in equine urine form adult horses varied widely between samples (8.5 +/- 6.5 nM). This variability was somewhat reduced when values were adjusted for dilutional effects using urine creatinine as an indicator (3.6 +/- 2.0 nanomoles/g creatinine). No significant differences were demonstrated between the means of EGF binding activity concentrations in clinically normal horses and horses affected by chronic laminitis.

Equine laminitis: a potential model of Raynaud's phenomenon.

Raynaud's phenomenon (RP) and equine laminitis in the horse are medical enigmas. Clinical and scientific data were compared to evaluate the degree of similarity that exists between these two peripheral vascular diseases. Data indicate that certain pathologic and pharmacologic aspects seem to have common features. Some of the correlations maybe due simply to both diseases having ischemia of the distal digits as a pathologic component. The exact etiology of the ischemia is not known for either disease. The results of this study suggest the hypothesis that RP and laminitis are the same disease in different species. This hypothesis can be tested more efficiently when the pathophysiology of both conditions is better documented. It is possible that comparative studies will promote advances in the understanding of both RP and laminitis. The fact that equine laminitis can be experimentally induced is of potential value in such future studies.

Effect of weight loading on the coronary band interstitial fluid pressure in horses.

Interstitial fluid pressures, as a possible function of limb load, were measured at 2 sites within the digital coronary dermis of both cranial digits in 10 standing horses. Fluid pressure changes and digital load measurements were simultaneously detected and recorded by use of, respectively, modified wick-in-needle and force plate transducers coupled to a microcomputer. Mean pressures, recorded at limb loads between 50 and 80 kg, were 2.29 +/- 3.17 mm of Hg at the toe and 2.49 +/- 5.91 mm of Hg at the heel. Mean pressures, recorded between 150 and 180 kg, were 5.01 +/- 5.23 mm of Hg at the toe and 1.28 +/- 7.69 mm of Hg at the heel. These data indicate that, in the static limb, no statistically significant change in interstitial fluid pressure occurs at loads up to 180 kg.

Epidermal growth factor-mediated effects on equine vascular smooth muscle cells.

Epidermal growth factor (EGF) receptor binding kinetics and EGF-mediated stimulation of DNA synthesis and cellular proliferation were studied in cultured vascular smooth muscle cells (VSMC) from the equine thoracic aorta. Binding studies, using murine 125I-labeled EGF, indicate the presence of a single class of high-affinity binding sites (apparent KD = 2.8 X 10(-11) M), with an estimated maximal binding capacity of 5,800 sites/cell. EGF stimulated [3H]thymidine uptake in confluent quiescent monolayers in a dose-dependent fashion, half-maximal stimulation occurring at 7.5 X 10(-11) M. Likewise, EGF-mediated cellular proliferation was dose dependent (50% effective dose = 5 X 10(-11) M) under reduced serum concentrations. Equine VSMC contain specific receptors for EGF, and EGF can stimulate DNA synthesis and proliferation in these cultured cells, which suggests that EGF may participate in the proliferative changes observed in equine distal digital peripheral vascular disease.