Estimation of acute fluid shifts using bioelectrical impedance analysis in horses

BACKGROUND: Multi-frequency bioelectrical impedance analysis (MF-BIA) has been used to evaluate extracellular fluid volume (ECFV), but not fluid fluxes associated with fluid or furosemide administration in horses. If able to detect acute changes in ECFV, MF-BIA would be useful in monitoring fluid therapy in horses. HYPOTHESIS: The purpose of this study was to evaluate the ability of MF-BIA to detect acute fluid compartment changes in horses. We hypothesized that MF-BIA would detect clinically relevant (10-20%) changes in ECFV. ANIMALS: Six healthy mares were used in the study. METHODS: This is an original experimental study. Mares were studied in 3 experiments: (1) crystalloid expansion of normally hydrated subjects, (2) furosemide-induced dehydration followed by crystalloid administration, and (3) acute blood loss followed by readministration of lost blood. MF-BIA measurements were made before, during, and after each fluid shift and compared to known changes in volume calculated based on the intravenous fluids that were administered in addition to urinary fluid losses. Mean errors between MF-BIA estimated change and known volume change were compared using nonparametric analysis of variance. Estimated ECFV pre- and post-fluid administration similarly were compared. The level of statistical significance was set at P < .05. RESULTS: Results of the study revealed a statistically significant change in ECFV and total body water during crystalloid expansion and dehydration. Statistically significant changes were not observed during blood loss and administration. Mean errors between MF-BIA results and measured net changes were small. CONCLUSIONS AND CLINICAL IMPORTANCE: MF-BIA represents a practical and accurate means of assessing acute fluid changes during dehydration and expansion of ECFV using isotonic crystalloids with potential clinical applications in equine critical care.     

Effect of endotoxin administration on body fluid compartments in the horse

Plasma volume, extracellular fluid volume (ECFV), and total body water (TBW) were measured before and after endotoxin (Escherichia coli) administration in 6 conscious adult horses. Evan's blue dye, sodium thiocyanate, and antipyrine were the test substances used to estimate plasma volume, ECFV, and TBW, respectively. Pharmacokinetic analysis of plasma concentration vs time was used to determine changes in body fluid compartments. The pathophysiologic effects of endotoxin were monitored by clinical evaluation, blood chemical changes, and blood gas determinations. All horses became dyspneic within 15 minutes of endotoxin administration and clinical signs of colic were evident 30 to 45 minutes after endotoxin administration. After endotoxin administration, serum glucose and creatinine concentrations were significantly (P less than 0.05) elevated, and all horses became hypoxic and developed marked metabolic acidosis, and plasma volume decreased approximately 15% (P less than 0.05). A significant change in ECFV or TBW during the 300-minute experimental period was not observed.     

Pharmacokinetics and clinical utility of sodium bromide (NaBr) as an estimator of extracellular fluid volume in horses

The purpose of this study was to describe the pharmacokinetics of bromide in horses and to evaluate the corrected bromide space as an indicator of extracellular fluid volume (ECFV) in horses after the administration of a single dose of bromide by intravenous infusion. Sodium bromide (30 mg/kg of body weight, IV) was administered to 6 clinically healthy mares over a period of 3 minutes. Blood samples were collected before infusion and at intervals between 0.5 hours and 53 days after infusion. Mean elimination half-life (harmonic mean) was 126 hours (5.2 days), clearance was 1.4 +/- 0.09 mL/(kg x h), area under the curve was 17,520 +/- 1,100 microg x h/mL. and volume of distribution (steady state) was 0.255 +/- 0.015 L/kg. The mean corrected bromide space was determined from the volume of distribution (steady state) and the serum concentrations of bromide at equilibration. Corrected bromide space, an estimate of ECFV, was 0.218 +/- 0.01 L/kg. The conclusion was made that ECFV of horses can be estimated by measuring bromide concentrations in a preinfusion serum sample and a sample obtained 5 hours after the administration of bromide.     

Use of a three-blood-sample plasma clearance technique to measure GFR in horses

Measurement of renal function in horses poses a particular challenge because plasma creatinine is influenced by muscle mass which is highly developed and variable between individuals, while conventional clearance methods involve potentially daunting problems, particularly urine collection and bladder washout. This paper provides data which enable technetium-diethyleneaminopentacetic acid (Tc-DTPA) clearance to be used to calculate glomerular filtration rate (GFR)/extracellular fluid volume (ECFV) as an expression of GFR in horses, as previously validated in humans, dogs and calves. Apart from being arguably a more physiological expression of GFR than using derivatives of body weight, the use of GFR/ECFV eliminates a source of delay and error, namely measurement of the injected dose, and offers the convenience of requiring only three blood samples. It therefore has advantages for both research and clinical applications.     

Comparison of inulin with urea as dilutional markers of bronchoalveolar lavage in healthy and heaves-affected horses

Solute analysis in bronchoalveolar lavage fluid involves the use of dilutional markers to correct for variable recovery of pulmonary epithelial lining fluid (PELF). Urea is the best characterised endogenous marker, whereas inulin appears to meet the requirements of an exogenous marker. In horses, the use of inulin has never been investigated and the impact of lower airway diseases such as heaves, on PELF recovery is unknown. In this study, five healthy and five heaves-affected horses underwent airway endoscopy and bronchoalveolar lavage. PELF recovery from bronchoalveolar lavage was calculated by the inulin and the urea method. The inulin method was compared to the urea method and differences between healthy and heaves-affected horses were analysed. From a technical and analytical point of view, inulin fulfilled the requirements of a marker of dilution as well as urea. When both healthy and heaves-affected horses groups were pooled together, PELF recovery calculated by the inulin method was significantly higher than by the urea method (6.43+/-4.08% versus 0.789+/-0.299%, P < 0.005). No significant differences were observed between healthy and heaves-affected horses, neither by the inulin nor by the urea method. Inulin did not present major advantages over urea, but the combined use of both markers can improve the standardisation of studies comparing PELF compounds, by providing upper limits (inulin dilution) and lower limits (urea dilution) of PELF recovery.     

Comparison of two indirect techniques for local delivery of a high dose of an antimicrobial in the distal portion of forelimbs of horses

OBJECTIVE: To compare isolated limb retrograde venous injection (ILRVI) and isolated limb infusion (ILI) for delivery of amikacin to the synovial fluid of the distal interphalangeal and metacarpophalangeal joints and to evaluate the efficacy of use of an Esmarch tourniquet in standing horses. ANIMALS: 6 healthy adult horses. PROCEDURES: Horses were randomly assigned in a crossover design. In ILRVI, the injection consisted of 1 g of amikacin diluted to a total volume of 60 mL administered during a 3-minute period. In ILI, the infusion consisted of 1 g of amikacin diluted to 40 mL administered during a 3-minute period followed by administration of boluses of diluent (82 mL total) to maintain vascular pressure. During ILI, the infusate and blood were circulated from the venous to the arterial circulation in 5-mL aliquots. Synovial fluid and serum samples were obtained to determine maximum amikacin concentrations and tourniquet leakage, respectively. RESULTS: Both techniques yielded synovial concentrations of amikacin > 10 times the minimum inhibitory concentration (MIC) for 90% of isolates (80 microg/mL) and > 10 times the MIC breakpoint (160 microg/mL) of amikacin-susceptible bacteria reported to cause septic arthritis in horses. These values were attained for both joints for both techniques. Esmarch tourniquets prevented detectable loss of amikacin to the systemic circulation for both techniques. CONCLUSIONS AND CLINICAL RELEVANCE: Both techniques reliably achieved synovial fluid concentrations of amikacin consistent with concentration-dependent killing for bacteria commonly encountered in horses with septic arthritis. Esmarch tourniquets were effective for both delivery techniques in standing horses.     

Glomerular filtration rate estimated by 3-sample plasma clearance of iohexol in 118 healthy dogs

BACKGROUND: Glomerular filtration rate (GFR) decreases in the aging human kidney, but limited data exist in dogs. HYPOTHESIS: There is an effect of age and body size on estimated GFR in healthy dogs. ANIMALS: One hundred and eighteen healthy dogs of various breeds, ages, and body weights presenting to 3 referral centers. METHODS: GFR was estimated in clinically healthy dogs between 1 and 14 years of age. GFR was estimated from the plasma clearance of iohexol, by a compartmental model and an empirical correction formula, normalized to body weight in kilograms or liters of extracellular fluid volume (ECFV). For data analysis, dogs were divided into body weight quartiles 1.8-12.4, 13.2-25.5, 25.7-31.6, and 32.0-70.3 kg. RESULTS: In the complete data set, there was no trend toward lower estimated GFR/kg or GFR/ECFV with increasing age. GFR decreased with age in dogs in the smallest weight quartile only. A significant negative linear relationship was detected between body weight and estimated GFR/kg and GFR/ECFV. Reference ranges in different weight quartiles were 1.54-4.25, 1.29-3.50, 0.95-3.36, and 1.12-3.39 mL/min/kg, respectively. Standardization to ECFV rather than kilogram body weight did not produce substantial changes in the relationships between GFR estimates and age or weight. CONCLUSIONS AND CLINICAL IMPORTANCE: Interpretation of GFR results for early diagnosis of renal failure should take into account the weight and the age of the patient for small dogs.     

Assessment of the accuracy of computed tomography for measurement of normal equine pituitary glands

OBJECTIVE: To describe the anatomic features of the pituitary gland region in horses via computed tomography (CT) and determine the accuracy of CT for estimating normal equine pituitary gland dimensions. ANIMALS: 25 adult horses with no clinical signs of pituitary disease. PROCEDURE: Transverse CT images and gross transverse tissue sections were compared in 2 horses. Contrast-enhanced CT of the pituitary gland region was performed postmortem in 23 horses with 4 slice thickness and interval settings (10-mm contiguous or overlapping slices and 4-mm contiguous or overlapping slices). Gross and CT estimates of pituitary gland dimensions were compared via ANOVA. Accuracy of CT estimates was calculated with gross pituitary gland measurements as the known value. RESULTS: Pituitary glands were located between the temporomandibular joints and had contrast enhancement. Mean gross dimensions were length, 2.11 cm; width, 2.16 cm; height, 0.98 cm; and volume, 2.66 cm3. Gross measurements and CT estimates of pituitary gland length from 10-mm contiguous and overlapping slices did not differ. Gross measurements and CT estimates of pituitary gland width from 4-mm contiguous and overlapping slices did not differ. Estimates of height and volume from all CT techniques differed from gross measurements. Accuracies for CT estimates were length, 88 to 99%; width, 81 to 92%; height, 58 to 71%; and volume, 43 to 55%. CONCLUSIONS AND CLINICAL RELEVANCE: Accuracy of estimates of pituitary gland dimension in horses varied with CT scanning technique; via CT estimates of length and width of glands were more accurate than estimates of height or volume.     

Effects of abdominocentesis technique on peritoneal fluid and clinical variables in horses

Eleven healthy horses underwent 5 repeated abdominocenteses, with either a sharp‐tipped spinal needle or a blunt‐tipped teat cannula to investigate possible differences in success rate, sample volume, depth at which a sample was obtained, length of procedure, complications and cytological variables. Variables were analysed with a repeated‐measures ANOVA or Fisher's exact test (α = 0.05). Success rate, sample volume, length of procedure, occurrence of haemorrhage during the procedure and incidence of grossly visible blood contamination were not different between techniques or over time. Depth at which samples were obtained was greater using a cannula than a needle (P<0.02), and samples were obtained with either technique at a greater depth than abdominal wall thickness assessed via ultrasound (P<0.014). Peritoneal fluid total and differential nucleated cell counts, and total protein concentration did not differ between techniques or over time. Red blood cell count in the least blood contaminated fraction of each sample was not affected by time, but it was lower after needle abdominocentesis than after cannula abdominocentesis (P = 0.04). Swelling of abdominocentesis sites increased with both techniques over time (P<0.05) and was more severe in horses undergoing cannula abdominocentesis (P<0.05). Enterocentesis occurred with a spinal needle in one horse, but no subsequent complications were noted. Both techniques appear to be safe and reliable for abdominocentesis in healthy horses. Using a blunt‐tipped cannula, as opposed to a needle, is likely to result in greater subcutaneous swelling. Both the cannula and needle must be long enough to penetrate well beyond the thickness of the abdominal wall to achieve successful peritoneal fluid collection.     

IOHEXOL PLASMA CLEARANCE IN HEALTHY DOGS AND CATS

Iohexol plasma clearance as a measure of glomerular filtration was determined in 31 dogs and 19 cats after an intravenous (i.v.) bolus injection. All animals were healthy and privately owned. Serial blood samples were taken before and up to 4 h after tracer injection. Iohexol plasma concentration was determined using X-ray fluorescence. A plasma tracer elimination curve was generated and clearance was calculated by dividing the injected dose by the area under the curve estimated using a two-compartment pharmacological model. Clearance was normalized to body weight (BW), body surface area (BSA), and extracellular fluid volume (ECFV). Mean, SD, and coefficient of variation of plasma clearance, before and after normalization, were calculated. Linear regression analyses were performed between body size and normalized plasma clearances. No significant linear relation was found between BSA and clearance normalized to BSA in dogs, and between BSA, BW, ECFV and clearance normalized to BSA, BW, and ECFV in cats. The optimal method for normalization of iohexol plasma clearance in dogs was by using BSA. In cats, all three methods tested were considered satisfactory. Normalization to BSA appears to be superior to normalization to BW and ECFV in dogs, and can be recommended for clinical use.     

Concentrations of serum amyloid A in serum and synovial fluid from healthy horses and horses with joint disease

OBJECTIVE: To determine serum amyloid A (SAA) concentrations in serum and synovial fluid from healthy horses and horses with joint disease and assess the effect of repeated arthrocentesis on SAA concentrations in synovial fluid. Animals-10 healthy horses and 21 horses with various types of joint disease. PROCEDURES: Serum and synovial fluid samples were obtained from each horse. In 5 of the 10 healthy horses, arthrocentesis was repeated 9 times. Concentrations of SAA were determined via immunoturbidometry. RESULTS: Serum and synovial fluid SAA concentrations were less than the assay detection limit in healthy horses and did not change in response to repeated arthrocentesis. Synovial fluid SAA concentrations were significantly higher in horses with suspected bacterial joint contamination or infectious arthritis, or tenovaginitis than in healthy controls, and serum concentrations were significantly higher in horses with infectious conditions than in the other groups. Neither serum nor synovial fluid SAA concentrations in horses with low-inflammation joint conditions differed significantly from those in healthy controls. Concentrations of SAA and total protein in synovial fluid were significantly correlated. CONCLUSIONS AND CLINICAL RELEVANCE: Synovial fluid SAA concentration was a good marker of infectious arthritis and tenovaginitis and appeared to reflect changes in inflammatory activity. The advantages of use of SAA as a marker include the ease and speed of measurement and the fact that concentrations in synovial fluid were not influenced by repeated arthrocentesis in healthy horses. Further study of the SAA response in osteoarthritic joints to assess its usefulness in diagnosis and monitoring of osteoarthritis is warranted.     

Assessment of body fat in the pony: part II. Validation of the deuterium oxide dilution technique for the measurement of body fat

Reasons for performing the study: Excessive accumulations or depletions of body fat have been associated with increased morbidity and mortality in horses and ponies. An objective, minimally‐invasive method to accurately quantify body fat in living animals is required to aid nutritional management and define welfare/performance limits. Objectives: To compare deuterium oxide (D₂O) dilution‐derived estimates of total body water (TBW) and body fat with values obtained by ‘gold standard’ proximate analysis and cadaver dissection. Hypothesis: D₂O dilution offers a valid method for the determination of TBW and body fat in equids. Methods: Seven mature (mean ± s.e. 13 ± 3 years, 212 ± 14 kg, body condition scores 1.25–7/9), healthy, Welsh Mountain pony mares, destined for euthanasia (for nonresearch purposes) were used. Blood samples were collected before and 4 h after D₂O (0.11–0.13 g/kg bwt, 99.8 atom percent excess) administration. Plasma was analysed by gas isotope ratio mass spectrometry following filtration and zinc reduction. After euthanasia, white adipose tissue (WAT) mass was recorded before all body tissues were analysed by proximate chemical analyses. Results: D₂O‐derived estimates of TBW and body fat were strongly associated with proximate analysis‐ and dissection‐derived values (all r²>0.97, P≤0.0001). Bland‐Altman analyses demonstrated good agreements between methods. D₂O dilution slightly overestimated TBW (0.79%, limits of agreement (LoA) ‐3.75–2.17%) and underestimated total body lipid (1.78%, LoA ‐0.59–4.15%) and dissected WAT (0.72%, LoA ‐2.77–4.21%). Conclusions and potential relevance: This study provides the first validation of the D₂O dilution method for the minimally‐invasive, accurate, repeatable and objective measurement of body water and fat in living equids.     

Concentrations of gentamicin in serum and bronchial lavage fluid after intravenous and aerosol administration of gentamicin to horses

OBJECTIVE: To compare concentrations of gentamicin in serum and bronchial lavage fluid after IV and aerosol administration of gentamicin to horses. ANIMALS: 9 healthy adult horses. PROCEDURE: Gentamicin was administered by aerosolization (20 ml of gentamicin solution [50 mg/ml]) and IV injection (6.6 mg of gentamicin/kg of body weight) to each horse, with a minimum of 2 weeks between treatments. Samples of pulmonary epithelial lining fluid were collected by small volume (30 ml) bronchial lavage 0.5, 4, 8, and 24 hours after gentamicin administration. Serum samples were obtained at the same times. All samples were analyzed for gentamicin concentration, and cytologic examinations were performed on aliquots of bronchial lavage fluid collected at 0.5, 8, and 24 hours. RESULTS: Gentamicin concentrations in bronchial lavage fluid were significantly greater 0.5, 4, and 8 hours after aerosol administration, whereas serum concentrations were significantly less at all times after aerosol administration, compared with IV administration. Neutrophil counts in bronchial lavage fluid increased from 0.5 to 24 hours, regardless of route of gentamicin administration. CONCLUSIONS AND CLINICAL RELEVANCE: Aerosol administration of gentamicin to healthy horses resulted in gentamicin concentrations in bronchial fluid that were significantly greater than those obtained after IV administration. A mild inflammatory cell response was associated with aerosol delivery of gentamicin and repeated bronchial lavage. Aerosol administration of gentamicin may have clinical use in the treatment of bacterial bronchopneumonia in horses.     

Concentrations of substance P and prostaglandin E2 in synovial fluid of normal and abnormal joints of horses

OBJECTIVE: To correlate substance P content of synovial fluid with prostaglandin E2 content, radiographic evidence of joint abnormality, and anatomic location of the joint for normal and osteoarthritic joints of horses. SAMPLE POPULATION: Synovial fluid from 46 normal joints in 21 horses and 16 osteoarthritic joints in 10 horses. PROCEDURE: Normal and osteoarthritic joints were identified by clinical and radiographic examination, by response to nerve blocks, during scintigraphy or surgery, or by clinicopathologic evaluation. Substance P and prostaglandin E2 contents of synovial fluid were determined by radioimmunoassay. Radio-graphs of joints were assigned a numeric score reflecting severity of lesions. Joints were assigned a numeric score reflecting anatomic location. RESULTS: Median concentrations of substance P and prostaglandin E2 were significantly increased in osteoarthritic joints, compared with normal joints. A significant correlation was found between concentrations of substance P and prostaglandin E2 in synovial fluid, but a correlation was not detected between substance P concentration in synovial fluid and anatomic location of the joint or between radiographic scores of osteoarthritic joints and concentrations of substance P or prostaglandin E2. CONCLUSIONS AND CLINICAL RELEVANCE: A correlation existed between concentrations of substance P and prostaglandin E2 in synovial fluid obtained from normal and osteoarthritic joints. However, content of substance P in synovial fluid cannot be predicted by the radiographic appearance of the joint or its anatomic location. Substance P and prostaglandin E2 may share an important and related role in the etiopathogenesis of osteoarthritis, lending credence to the importance of neurogenic inflammation in horses.     

Evaluation of peritoneal fluid pH, glucose concentration, and lactate dehydrogenase activity for detection of septic peritonitis in horses

OBJECTIVE: To determine whether peritoneal fluid pH, glucose concentration, and lactate dehydrogenase activity can be used to differentiate horses with septic peritonitis from those with nonseptic peritonitis. DESIGN: Prospective study. ANIMALS: 46 horses, including 10 healthy horses, 15 horses with septic peritonitis, and 21 horses with nonseptic peritonitis. PROCEDURE: Peritoneal fluid and blood samples were analyzed for pH, glucose concentration, and lactate dehydrogenase activity. Complete blood cell counts were performed, and peritoneal fluid samples were submitted for bacterial culture. RESULTS: Horses with septic peritonitis had significantly lower peritoneal fluid pH and glucose concentrations than horses with nonseptic peritonitis and healthy horses. Compared with other tests, serum-to-peritoneal fluid glucose concentration differences > 50 mg/dl had the highest diagnostic use for detection of septic peritonitis. Peritoneal fluid pH < 7.3, glucose concentration < 30 mg/dl, and fibrinogen concentration > 200 mg/dl were also highly indicative of septic peritonitis. CLINICAL IMPLICATIONS: Peritoneal fluid pH and glucose concentration can be used to assist in the identification of horses with septic peritonitis. These measurements can provide an early indication of sepsis, especially if cytologic evaluation of peritoneal fluid is unavailable or results are equivocal and peritoneal fluid bacterial culture results are pending.     

Comparative disposition of pharmacologic markers for cytochrome P-450 mediated metabolism, glomerular filtration rate, and extracellular and total body fluid volume of Greyhound and Beagle dogs

The purpose of the study was to compare the disposition of pharmacologic markers for cytochrome P-450 (CYP) metabolism, glomerular filtration rate (GFR), and extracellular (ECFV) and total body fluid volumes (TBFV) of Greyhounds and Beagles. Six healthy Greyhound and six healthy Beagle dogs were studied. Antipyrine, a marker for CYP metabolism and TBFV, and inulin, a marker for the GFR and ECFV, were administered i.v. Samples were collected at predetermined times and plasma was analyzed by validated high-pressure liquid chromatography (HPLC) methods. There were no differences in the disposition or pharmacokinetic parameters for inulin between the dog breeds. However, the clearance of antipyrine (mean = 8.33 mL/min/kg) in Greyhounds was significantly slower than Beagles (13.42 mL/min/kg, P = 0.004). The volume of distribution of antipyrine was significantly larger in Greyhounds (0.789 L/kg) than in Beagles (0.644 L/kg, P = 0.01). The half-life of antipyrine was significantly longer in Greyhounds (1.09 h) compared with Beagles (0.55 h, P = 0.002). The in vitro plasma protein binding of antipyrine was significantly less in Greyhounds (28%) compared with Beagles (40.3%, P = 0.008). Greyhounds exhibited significantly slower CYP metabolism, higher TBFV, and lower in vitro protein binding of antipyrine compared with Beagles. No differences in GFR or ECFV were found.     

Characterization of lipoproteins in cerebrospinal fluid of mares during pregnancy and lactation

OBJECTIVE: To measure apolipoproteins in cerebrospinal fluid (CSF) from healthy mares and to determine whether CSF concentrations of apolipoproteins change during pregnancy and lactation. ANIMALS: 5 healthy pregnant mares. PROCEDURE: 2 sets of CSF samples were obtained; initial samples were obtained 10 to 30 days before parturition (mean, 18 days; median, 17 days), and second samples were obtained 19 to 26 days after parturition (mean, 23 days; median, 23 days). Cerebrospinal fluid was collected from the lumbosacral subarachnoid space of standing horses by use of routine collection techniques. Cerebrospinal fluid cholesterol concentrations were measured by use of a sensitive enzymatic assay. Ultracentrifugal fractions of CSF lipoproteins were characterized by determining the distribution of apolipoproteins, using polyacrylamide gel electrophoresis. RESULTS: Analyses of isolated ultracentrifugal fractions by polyacrylamide gel electrophoresis revealed 2 apolipoproteins, with the expected molecular weights for apolipoprotein E and apolipoprotein A-I. No significant differences were observed between pre- and postpartum values in mares. The concentration of cholesterol in CSF fluid of mares was comparable to values reported in other mammals. CONCLUSIONS AND CLINICAL RELEVANCE: Apolipoprotein E in CSF of horses is a major apolipoprotein associated with high-density lipoproteins, which is similar to findings in other mammals. Additional characterization of the role of apolipoproteins in mammalian CSF may provide critical insight into various degenerative neurologic disease processes.     

Immunohistochemical determination of the expression of endothelin receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease

OBJECTIVE: To immunohistochemically determine the expression of endothelin (ET) receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease (SPAOPD). SAMPLE POPULATION: Tissue specimens obtained from 8 healthy and 8 SPAOPD-affected horses. PROCEDURE: Horses were examined and assigned to healthy and SPAOPD groups. Horses were then euthanatized, and tissue specimens containing bronchi of approximately 4 to 8 mm in diameter were immediately collected from all lung lobes, fixed in zinc-formalin solution for 12 hours, and embedded in paraffin. Polyclonal primary antibodies against ET-A or ET-B receptors at a dilution of 1:200 and biotinylated IgG secondary antibodies were applied to tissue sections, followed by the addition of an avidin-biotin immunoperoxidase complex. Photographs of the stained slides were digitally recorded and analyzed by use of image analysis software to determine the intensity of staining. Two-way ANOVA was used for statistical analysis. RESULTS: The left diaphragmatic lung lobe of SPAOPD-affected horses had a significantly greater area of bronchial smooth muscle that immunostained for ET-A, compared with that for healthy horses. All lung lobes of SPAOPD-affected horses, except for the right diaphragmatic lobe, had significantly greater staining for ET-B receptors in bronchial smooth muscle, compared with results for healthy horses. CONCLUSIONS AND CLINICAL RELEVANCE: This study revealed overexpression of ET-A and, in particular, ETB receptors in the bronchial smooth muscle of SPAOPD-affected horses, which suggested upregulation of these receptors. These findings improve our understanding of the role of ET-1 in the pathogenesis of SPAOPD.     

Reduction of renal function in exercising horses

Glomerular filtration rate (GFR) and extracellular volume (ECFV) were measured before, during and after treadmill exercise in 5 trained Thoroughbred horses (mean weight 483 kg). GFR/ECFV was determined by plasma disappearance of Tc-DTPA and ECFV was measured independently as thiocyanate space. Resting GFR averaged 1.6 l/min (3.3 ml/kg/min) and fell by over 40% during exercise, moreover the fall was severe even during the first walk, prior to trotting. The results suggest that rather than being protected, GFR is allowed to fall, even with mild exercise and that this is probably an adaptation to allow greater perfusion of muscle and skin. In man, GFR appears to be more resistant to the effects of exercise but it is hard to compare intensity of exercise between such different species.     

Use of volumetric capnography to identify pulmonary dysfunction in horses with and without clinically apparent recurrent airway obstruction

OBJECTIVE: To investigate whether volumetric capnography indices could be used to differentiate between horses without recurrent airway obstruction (RAO) and horses with RAO that were in clinical remission or that had clinically apparent RAO. ANIMALS: 70 adult Swiss Warmblood horses (20 used for pleasure riding and 50 used for dressage or show jumping). PROCEDURE: Horses were allocated to 4 groups on the basis of history, clinical signs, results of endoscopy, and cytologic findings (group 1, 21 healthy horses; group 2, 22 horses with RAO that were in remission; group 3, 16 horses with mild RAO; group 4, 11 horses with exacerbated RAO). Expiratory volume and CO2 curves were recorded by use of a computerized ultrasonic spirometer. Volumetric capnograms were plotted, and derived indices were calculated. RESULTS: Dead-space volume (VD) was calculated by use of the Bohr equation (VD(Bohr)) and for physiologic VD (VD(phys)). Ratios for VD(Bohr) to expiratory tidal volume (VT) and VD(phys) to V(T) as well as an index of effective CO2 elimination were significantly different among groups of horses. Age and use of the horses also significantly affected volumetric capnography indices. CONCLUSIONS AND CLINICAL RELEVANCE: Ratios of VD(Bohr) to VT and VD(phys) to VT as well as an index of effective CO2 elimination were sufficiently sensitive measures to distinguish between healthy horses and horses with RAO in remission. To optimize the ability of volumetric capnography indices to differentiate among horses in heterogeneous populations, it is important to account for effects of age and specific use of the horses.