Acute hemolytic anemia after oral administration of L-tryptophan in ponies

The hematologic and pathologic effects of orally administered L-tryptophan and indoleactic acid and of L-tryptophan administered IV were studied in ponies. Sixteen adult Shetland ponies were allotted into 4 experimental groups. Group 1 consisted of 5 ponies (1-5) given 0.6 g of tryptophan/kg of body weight in a water slurry via stomach tube. Group 2 included 4 ponies (6-9) given 0.35 g of tryptophan/kg orally. Group-3 ponies (10-13) were given 0.35 g of indoleacetic acid/kg orally. Group 4 consisted of 3 ponies (14-16) given a single 4-hour IV infusion of 0.1 g of tryptophan/kg. Restlessness, increased respiratory rate, hemolysis, and hemoglobinuria were detected in 4 of the 5 group-1 ponies. Only pony 7 in group 2 developed hemolysis, hemoglobinuria, and a significant increase in respiratory rate. Renal pathologic lesions, consistent with hemoglobinuric nephrosis, were seen in ponies 2, 4, 5, and 7. Bronchiolar degeneration was evident in 4 of 9 ponies given tryptophan orally. The importance of these respiratory lesions was unknown. Clinical or pathologic abnormalities were not noticed in the ponies of groups 3 and 4. Mean plasma tryptophan values increased significantly in groups 1 and 2 at 6 hours after dosing. A second peak of tryptophan was detected in both groups at 12 hours. Values returned to predose values by 48 hours. Plasma indole and 3-methylindole concentrations were detectable in only 2 ponies (4 and 7). In vitro incubations of cecal fluid from ponies 6, 8, and 9 yielded a percentage conversion of tryptophan to indole of 16.75%, 5.84%, and 7.96%, respectively. 3-Methylindole was not produced. These results suggested that indole was the major metabolite of orally administered tryptophan in these ponies.     

Acute hemolytic anemia induced by oral administration of indole in ponies

Eight ponies were allotted to 2 groups of 4. Group-1 ponies (1-4) were given 0.2 g of indole/kg of body weight orally and group-2 ponies (5 to 8) were given 0.1 g of indole/kg. Various physical, hematologic, and physiologic measurements were obtained after administration of indole. Intravascular hemolysis and hemoglobinuria were detected in both groups within 24 hours of dosing. Hemolysis was reflected by decreases in PCV, hemoglobin concentration, and RBC count, and an increase in indirect bilirubin. Erythrocyte fragility appeared to increase in both groups at 8 hours after dosing and peaked at 16 hours after dosing. At 72 hours after dosing, the RBC fragility value was less than predose measurements. Heinz body formation was noticed in group-2 ponies, but not in group 1. Plasma indole concentrations increased in both groups from the nondetectable predose concentrations. Group-1 values were 203% of group-2 values. In group 2, plasma indole was nondetectable by 12 hours, whereas low concentrations could still be measured in the group-1 ponies at 24 hours. Ponies in group 1 died or were euthanatized between 24 and 72 hours after dosing, whereas group-2 ponies were euthanatized between 48 and 120 hours. At necropsy, all body fat, mucous membranes, and elastic tissue were stained yellow. Hemoglobinuric nephrosis was the most prominent microscopic lesion. Results of this study indicated that indole, a metabolite of the amino acid tryptophan, causes acute intravascular hemolysis in ponies.     

Gastrointestinal ultrasonography in normal adult ponies

The objective of this study was to determine the characteristics based on ultrasonographic examination of the stomach, duodenum, jejunum, cecum, and peritoneal fluid in normal adult ponies. Abdominal ultrasonographic examination was performed in nine unsedated standing ponies. The duodenum was examined at three sites and the jejunum in 12 regions. Wall thickness, contractility, distention, and luminal contents were recorded. Stomach wall thickness and location, cecal wall thickness, and peritoneal fluid location and character were recorded. Statistical analysis was performed. Wall thicknesses (in cm) were 0.431 +/- 0.069 for the stomach, 0.188 +/- 0.033 for the duodenum (at all sites), 0.195 +/- 0.031 for the jejunum (at all regions), and 0.179 +/- 0.031 for the cecum. Duodenal contractions per minute were 3.78 +/- 1.10. The stomach spanned 5.14 +/- 0.9 intercostal spaces, with the 8th intercostal space being the most cranial and the 15th intercostal space being the most caudal space through which the stomach was identified. It was possible to identify the jejunum in all ponies dorsal to the left dorsal colon and from the ventral abdominal wall. Peritoneal fluid was identified in six ponies. Peritoneal fluid was usually seen transiently and most commonly in the ventral aspect of the abdominal cavity or around the duodenum. Overall, the ponie's abdominal ultrasonographic examinations revealed wall thicknesses that were less than the published normal ranges for horses. It appears that ponies may have increased duodenal contractility than horses and that the conformation of ponies may change the locations for imaging the stomach.     

Effect of furosemide administration on systemic circulation of ponies during severe exercise

Systemic distribution of blood flow was studied in 11 healthy adult grade ponies, using radionuclide-labeled microspheres (15 micron diameter) that were injected into the left ventricle. Measurements were made at rest, during severe exercise (SE) without furosemide, as well as during SE at 10 minutes and 120 minutes after furosemide administration (1.0 mg/kg, IV). During SE, heart rate, cardiac output, mean aortic pressure, and whole body O2 consumption were 220 +/- 4 beats/min, 720 +/- 44 ml/min/kg, 169 +/- 4 mm of Hg, and 126 +/- 9 ml of O2/min/kg, respectively. With SE performed after furosemide administration, mean aortic pressure decreased from prefurosemide SE value (P less than 0.05), but heart rate, cardiac output, and whole body O2 consumption remained similar to values during SE without furosemide. During SE, blood flow to cerebellar gray matter, pons, and medulla oblongata increased despite marked hypocapnia, but in other regions of the brain, blood flow was unchanged. As arterial O2 content increased by 58% with SE, O2 delivery to all brain regions increased. With SE, adrenal gland blood flow increased, but intense vasoconstriction in the kidneys, spleen, pancreas, small intestine, and colon caused blood flow to plummet. During SE, blood flow in the diaphragm, gluteus medius, biceps femoris (muscles of propulsion), and triceps brachii muscles increased to a similar level, indicating that metabolic requirements of the diaphragm during exercise may not be less than those of other vigorously contracting muscles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chronic oral therapy with enalapril in normal ponies

Enalapril is an angiotensin converting enzyme (ACE) inhibitor that is frequently used in human, feline and canine patients with cardiac disease. Its use has been associated with impotence in human patients. The purpose of this study was to evaluate if enalapril (0.5 mg/kg PO, q24h) is likely to alter behavior in stallions and to assess its effect on ACE activity at the standard dose used in dogs and cats. Twelve pony stallions were evaluated by physical examination and echocardiography followed by treatment with enalapril (n = 6) or placebo (n = 6) for 2 months. After one month, blood was drawn and stored to evaluate ACE activity in the 2 groups. At the end of the study, repeat physical examination and echocardiography were performed. Physical examination, echocardiographic indices, and reproductive performance were unchanged and there was no suppression of ACE activity. Results of this study suggest that enalapril (0.5 mg/kg PO, q24h) is either poorly absorbed in the horse or is inadequately converted to the active form of the drug, enalaprilat.     

Pharmacokinetics of gallium nitrate after oral administration in adult horses--pilot study

Pollina, G. F., Zagotto, G., Maritan, P., Iacopetti, I., Busetto, R Pharmacokinetics of gallium nitrate after oral administration in adult horses – pilot study. J. vet. Pharmacol. Therap.  35 , 489–494. Gallium (Ga), a metal in group IIIA of the periodic table, has shown a remarkable activity against bone resorption and could therefore possibly prove useful in the treatment of certain diseases in sport horses, for example navicular disease. The aim of this study was to gain more information concerning the kinetics of Ga after oral administration of gallium nitrate (GaN) in adult horses. Six horses received a single dose of 10 mg/kg of GaN mixed with the food ration. Absorption was slow (T_max = 10 ± 3 h, T_½abs = 2 ± 0.8 h), and a C_max of 26 ± 11 μg/L was achieved. Excretion followed a one‐phase elimination model, with a long half‐life (T_½el = 52 ± 14 h). By means of a mathematical model, we estimated that the plasmatic levels should reach 93 μg/L (1.33 μm ) at steady state, following the repeated daily administration of 10 mg/kg of GaN. A three times lower concentration has been demonstrated as effective in inhibiting the osteolytic activity of osteoclasts in vitro. The results of this study suggest that the administration of oral GaN at a rate of 10 mg/kg per day may be considered for future clinical studies.     

Multiple oral dosing of valacyclovir in horses and ponies

The aim of the current study was to investigate whether multiple oral dosing of valacyclovir could result in plasma concentrations exceeding the EC₅₀-value of acyclovir against equine herpesvirus 1 (EHV1) during the majority of the treatment period. Additionally, we wanted to determine the concentration of acyclovir in nasal mucus and cerebrospinal fluid (CSF). Valacyclovir was administered to four horses and two ponies, three times daily, at a dosage of 40 mg/kg, for four consecutive days. Blood was collected prior to each administration and 1 h after dosing. Nasal mucus samples and CSF were collected once during treatment; 1 h after the last administration. This dosage regimen resulted in plasma concentrations that were higher than the EC₅₀-value of 1.7 μg/mL, i.e. EC₅₀ of an isolate highly susceptible to acyclovir, for 80% of the treatment period; and higher than the EC₅₀-value of 3.0 μg/mL, i.e. EC₅₀ of an isolate less susceptible to acyclovir, for 60% of the treatment period. Concentration in nasal mucus samples and CSF was 0.36–1.17 μg/mL and 0.11–0.23 μg/mL, respectively. This study illustrates that multiple dosing of valacyclovir may result in a therapeutic benefit as plasma concentrations could be maintained above the EC₅₀-value of acyclovir against EHV1 for more than 50% of the treatment period. Acyclovir could be detected in both nasal mucus samples and CSF. However, these concentrations were lower than the EC₅₀.     

Pathophysiology of excessive iron storage in mynah birds

Hepatic iron overload was diagnosed in young and adult Rothschild's mynah birds. By light microscopic evaluation, it was determined that there was progressive accumulation of iron pigment with age in the hepatocytes. The Kupffer's cells of the liver, the reticuloendothelial cells of the spleen, and the phagocytic cells in the small intestine were negative for Prussian blue (iron) pigments. These observations suggested that diet had only a minor influence on the iron distribution in the mynah birds. This was confirmed by comparing iron distribution in the liver and spleen of mynah birds with that in other exotic birds receiving the same diet as the mynah birds. The syndrome of excessive iron overload in the mynah birds shared most of the important histopathologic characteristics with idiopathic (hereditary) hemochromatosis in human beings.     

Pharmacokinetics of maropitant citrate after oral administration of multiple doses in adult horses

The neurokinin-1 (NK-1) receptor antagonist, maropitant citrate, mitigates nausea and vomiting in dogs and cats. Nausea is poorly understood in horses, and clinical use of NK-1 receptor antagonists has not been reported. This study aimed to determine the pharmacokinetics and safety of maropitant after administration of multiple doses. We hypothesized that maropitant concentrations would be similar at steady state to those reported in dogs, with minimal adverse effects. Maropitant was administered at 4 mg/kg orally, once daily for 5 days in seven adult horses. Serial plasma maropitant concentrations were measured by liquid chromatography-mass spectrometry. Noncompartmental pharmacokinetic parameters were determined. The maximum, minimum, and average concentrations of maropitant achieved at steady state were 375.5 ± 200, 16.8 ± 7.7, and 73.5 ± 45.1 ng/ml, respectively. The terminal elimination half-life was 11.6 ± 1.4 hr, and the accumulation index was 1.3 ± 0.07. Heart rate decreased between Day 1 and Day 5 (p = .005), with three horses having heart rates of 20 beats per minute and atrioventricular block on Day 5. Pharmacokinetics of repeated maropitant administration suggests the drug could be considered for use in healthy horses. Further investigation on the clinical relevancy of its cardiac effects is warranted.     

Assessment of the minimum protein requirement of adult ponies

The aim of the present study was to determine the minimum protein requirement in adult ponies. Literature data from nitrogen-balance trials in adult ponies were used. In addition, nitrogen balance was measured in three mature ponies that were fed a low- and a high-protein diet according to a cross-over design. On the low-protein ration, which provided 1.1 g digestible crude protein (DCP)/kg(0.75) day, plasma albumin and total protein concentrations were non-significantly decreased by about 6% and group-mean nitrogen balance was negative. Using data from the present experiment and from six other studies, the sum of endogenous faecal and urinary nitrogen losses was calculated to be 166 +/- 37 mg/kg(0.75) day (mean +/- SE, n = 13). Subsequently, a minimum protein requirement of 1.7 g DCP/kg(0.75) day was calculated. The value for minimum protein requirement has low precision, which should be taken into account when setting the protein allowance of adult ponies.     

Effect of oral melatonin administration on sex hormone, prolactin, and thyroid hormone concentrations in adult dogs.

OBJECTIVE: To determine the effect of oral melatonin (MT) administration on serum concentrations of sex hormones, prolactin, and thyroxine in dogs. DESIGN: Prospective study. ANIMALS: 8 male and 8 female adult sexually intact dogs. PROCEDURE: 5 male and 5 female dogs were treated with MT (1.0 to 1.3 mg/kg [0.45 to 0.59 mg/lb] of body weight), PO, every 12 hours for 28 days; the other 6 dogs were used as controls. Blood samples were collected on days 0, 14, and 28, and serum concentrations of estradiol-17 beta, progesterone, testosterone, androstenedione, 17-hydroxyprogesterone (17-HP), dihydroepiandrostenedione sulfate (DHEAS), prolactin, and thyroxine were determined. On day 5, serum MT concentrations were measured before and periodically for up to 8 hours after MT administration in 4 treated dogs. RESULTS: Female dogs treated with MT had significant decreases in serum estradiol, testosterone, and DHEAS concentrations between days 0 and 28. Male dogs treated with MT had significant decreases in serum estradiol and 17-HP concentrations between days 0 and 28. Serum MT concentrations increased significantly after MT administration and remained high for at least 8 hours. Prolactin and thyroxine concentrations were unaffected by treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Melatonin is well absorbed following oral administration and may alter serum sex hormone concentrations.     

Effect of dietary fructans and dexamethasone administration on the insulin response of ponies predisposed to laminitis

OBJECTIVE: To determine whether pasture, and specifically the addition of fructan carbohydrate to the diet, induces exaggerated changes in serum insulin concentration in laminitispredisposed (LP) ponies, compared with ponies with no history of the condition, and also to determine insulin responses to the dexamethasone suppression test. DESIGN: Prospective study. ANIMALS: 10 LP and 11 control adult nonobese mixed-breed ponies. PROCEDURES: Insulin-modified IV glucose tolerance tests were performed (5 ponies/group). In diet studies, ponies were kept on pasture and then changed to a hay diet (10 ponies/group). Second, ponies were maintained on a basal hay diet (4 weeks) before being fed a hay diet supplemented with inulin (3 g/kg/d [1.4 g/lb/d]). Serum insulin and plasma glucose concentrations were analyzed before and after dietary changes. Serum cortisol and insulin concentrations were also measured in a standard dexamethasone suppression test. RESULTS: The LP ponies were insulin resistant (median insulin sensitivity of 0.27 x 10(4) L min(-1) mU(-1) in LP ponies, compared with 0.64 x 10(4) L min(-1) mU(-1) in control ponies). Median insulin concentration in LP ponies was significantly greater than that in control ponies at pasture, decreased in response to feeding hay, and was markedly increased (5.5-fold) following the feeding of inulin with hay. The LP ponies had a greater increase in serum insulin concentration at 19 hours after dexamethasone administration (median, 222.9 mU/L), compared with control ponies (45.6 mU/L). CONCLUSIONS AND CLINICAL RELEVANCE: Nonobese ponies predisposed to develop laminitis had compensated insulin resistance, and this phenotype was revealed by feeding plant fructan carbohydrate or by dexamethasone administration.     

Pharmacokinetics of acyclovir after single intravenous and oral administration to adult horses

The purpose of the study reported here was to describe the bioavailability and pharmacokinetics of acyclovir after intravenous and oral administration to horses. Six healthy adult horses were used in a randomized cross-over study with a 3 x 3 Latin square design. Three treatments were administered to each horse: 10 mg of injectable acyclovir/kg of body weight in 1 L of normal saline delivered as an infusion over 15 minutes; 10 mg of acyclovir/kg in tablets by nasogastric intubation; and 20 mg of acyclovir/kg in tablets by nasogastric intubation. A 2-week washout period was provided between each treatment. Serum samples were obtained for acyclovir assay using reversed-phase, high-performance liquid chromatography with fluorescence detection. Deproteinated serum was injected onto a C18 column, and elution occurred under isocratic conditions. The limit of quantification was 0.04 microg/mL. The assay exhibited suitable accuracy, precision, and recovery. The IV data were analyzed by a 3-compartment model, and oral data were analyzed noncompartmentally. Intragastric acyclovir administration at either dose was associated with high variability in serum acyclovir-time profiles, low Cmax, and poor bioavailability. The dosage of 20 mg/kg was associated with mean (+/- SD) Cmax of 0.19 +/- 0.10 microg/mL, and bioavailability was 2.8%. Inhibition of equine herpesvirus has been reported to require significantly higher acyclovir concentrations than those obtained here. The results of this study do not support a therapeutic benefit for the oral administration of acyclovir up to doses of 20 mg/kg.     

The effect of oral metformin on insulin sensitivity in insulin-resistant ponies

Metformin may be an effective therapeutic option for insulin-resistant (I-R) horses/ponies because, in humans, it reportedly enhances insulin sensitivity (SI) of peripheral tissues without stimulating insulin secretion. To determine the effect of metformin on insulin and glucose dynamics in I-R ponies, six ponies were studied in a cross-over design by Minimal Model analysis of a frequently-sampled intravenous glucose tolerance test (FSIGT). Metformin was administered at 15 mg/kg bodyweight (BW), orally, twice-daily, for 21 days to the metformin-treated group. The control group received a placebo. A FSIGT was conducted before and after treatment. The Minimal Model of glucose and insulin dynamics rendered indices describing SI, glucose effectiveness (Sg), acute insulin response to glucose (AIRg) and the disposition index (DI). The body condition score (BCS), BW and cresty neck score (CNS) were also assessed. There was no significant change in SI, Sg, AIRg, DI, BW, BCS or CNS in response to metformin, or over time in the control group. There were no measurable benefits of metformin on SI, consistent with recent work showing that the bioavailability of metformin in horses is poor, and chronic dosing may not achieve therapeutic blood concentrations. Alternatively, metformin may only be effective in obese ponies losing weight or with hyperglycaemia.