Neospora caninum causes serious disease in dogs, and it, or a similar parasite, is a major cause of abortion in cattle. Little is known about the susceptibility of this protozoan to antimicrobial agents. We studied several antimicrobial agents to determine which classes might have activity against this parasite. We also determined whether activity of such agents was coccidiocidal or coccidiostatic. A 2-day of treatment, monoclonal antibody-based enzyme immunoassay and a 5-day of treatment, cell culture flask (CCF), lesion-based assay were developed to examine the ability of test agents to inhibit tachyzoite multiplication. Seven sulfonamides were examined, with the following activities observed: sulfathiazole greater than or equal to sulfamethoxazole > sulfadiazine > sulfaquinoxaline greater than or equal to sulfamethazine > sulfadimethoxine > sulfamerazine. Dapsone, a sulfone, had little activity. Six dihydrofolate reductase/thymidylate synthase inhibitors were examined, with the following activities observed: piritrexim > pyrimethamine > ormetoprim > trimethoprim = diaveridine > methotrexate. Six ionophorous antibiotics were examined; lasalocid, maduramicin, monensin, narasin, and salinomycin had equivalent activities, but alborixin was toxic for host cells at the lowest concentration examined. Three macrolide antibiotics--azithromycin, clarithromycin, and erythromycin--were examined and had equivalent activities. Two tetracycline antibiotics, doxycycline and minocycline, were examined and had equivalent activities. Three lincosamide antibiotics were examined, with the following activities observed: clindamycin hydrochloride > clindamycin phosphate > lincomycin hydrochloride. Pentamidine and 6 of its analogs were examined, and only hexamidine and 1,4-Di[4-(2-imidazolinyl)-2-methoxy-phenoxylbutane had activity. Eight miscellaneous antiprotozoal agents were examined for activity. Amprolium, metronidazole, paromomycin, and roxarsone had little activity. Arprinocid, diclazuril, nitrofurazone, and robenidine had good activity. Eleven agents were examined in both assays, whereas 32 agents were examined in the CCF assay only. The enzyme immunoassay and CCF assay provided similar results for agents that rapidly killed tachyzoites. However, agents that inhibited development, but were not rapidly fatal for tachyzoites, had better activity in the CCF assay. Of the classes of agents examined, the dihydrofolate reductase/thymidylate synthase inhibitors, 2 of the 6 pentamidine analogs, and the ionophores were coccidiocidal and the sulfonamides, macrolides, tetracyclines, and lincosamides were coccidiostatic. Of the miscellaneous agents examined, arprinocid, nitrofurazone, and robenidine were coccidiocidal and diclazuril was coccidiostatic.

Seven horses (4 anesthetized and 3 awake) and 2 ponies (anesthetized) were studied to evaluate the high sensitivity of the pulmonary circulation of the horse to various blood-borne particles, and to establish the presence of intravascular macrophages in the lung. Pulmonary and systemic pressures and cardiac output before and during particle injection were measured in some animals. An anesthetized foal had a large increase in pulmonary arterial pressure (32 and 34 mm of Hg) within 1 minute of iv administration of small test doses of radioactively labeled liposomes (2.5 micromoles/kg of body weight) or a 1% suspension of blue pigment (0.3 ml/kg), respectively. Quantitative real-time gamma camera imaging of the foal revealed high retention of the labeled liposomes during the first pass through the lungs; retention persisted throughout the experiment. Postmortem analysis revealed 55 and 47% lung retention of liposomes and blue pigment, respectively. The 2 anesthetized ponies had increased pulmonary artery pressure of 34 +/- 7 mm of Hg, decreased cardiac output, and 42% lung retention after administration of 1% blue pigment (0.2 ml/kg), whereas 3 awake horses had increased pressure of 28 +/- 9 mm of Hg after 1.8 X 10(8) (1.8-micromoles-diameter) latex microspheres/kg. None of the injected particles caused vascular obstruction, and they do not cause pulmonary vascular reactivity in species that lack pulmonary intravascular macrophages. Finally, 3 horses (1 anesthetized and 2 awake) were infused iv with small doses of the blue pigment, and their lungs were perfusion-fixed to identify specific labeling of the pulmonary intravascular macrophages. These cells were fully differentiated macrophages, contained blue pigment in phagocytes, and were tightly adherent to the pulmonary capillary endothelium. At this time, horses (order Perissodactyla) are the only species outside the mammalian order Artiodactyla (sheep, pig, cattle) documented to have reactive intravascular macrophages. Compared with other species, low doses of particles induced marked hemodynamic responses; horses appear to be more sensitive to IV administered particles than are other species studied.

The hypothesis that equine laminitis is caused by thrombosis of vessels in the laminar corium (dermis) was investigated. Hemostatic alterations were evaluated by determining platelet count, platelet survival, platelet adhesiveness to vascular subendothelium, activated clotting time, and whole blood recalcification time. Thrombosis of vessels in the hoof wall was evaluated by scintigraphic studies of the hoof wall after administration of indium-111 ((111)In)- labeled platelets, contrast arteriography, and histologic examination. Platelet count remained constant before and at the onset of lameness; however, survival of (111)In-labeled platelets was shortened. Scintigraphy of affected feet revealed accumulation of (111)In-labeled platelets distal to the coronary band. Arteriography of disarticulated saline-perfused feet revealed marked reduction in blood supply to affected hooves. Histologic examination of the laminar dermis disclosed variable numbers of microthrombi in dermal veins of affected feet from 3 of 4 ponies with laminitis. Whole blood recalcification time was shortened at 8 hours after administration of carbohydrate and was prolonged at the onset of laminitis. Activated clotting time was prolonged at 32 hours after carbohydrate administration and at the onset of lameness. Plasma endotoxin-like activity was detected in 1 of 4 affected ponies. These data confirm that microvascular thrombosis existed at the onset of lameness in ponies with carbohydrate-induced laminitis and indicate that systemic coagulopathy may have preceded development of thrombosis.

A solid-phase ELISA to detect antibodies bound to the surface of canine platelets (platelet-bound antibodies) is described. Using this assay, the effect of anticoagulant and storage time of anticoagulant blood on the concentration of antibodies bound to the surface of platelets from clinically normal dogs was investigated. Blood from 3 clinically normal dogs was anticoagulated with acid citrate dextrose, Na3 citrate, and aqueous K3 EDTA and stored on ice for up to 48 hours. Platelet-bound antibody concentration was measured on platelets isolated from anticoagulated blood immediately after venipuncture and subsequent to storage of blood for 24 and 48 hours. Differences in platelet-bound antibody concentrations were investigated among dogs, anticoagulants, and storage times by ANOVA and Bonferroni pair-wise comparison of means. There was no effect of dog on platelet-bound antibody concentration. The effect of time was significant (P < 0.0001), with higher concentration of platelet-bound antibodies detected with increasing storage time. Effect of anticoagulant on platelet-bound antibody concentration was not statistically significant; however, there was a trend to increasing concentration of antibodies bound to platelets isolated from Na3 citrate- and K3 EDTA-anticoagulated blood. Moreover, there was significant (P = 0.02) interaction between anticoagulant and time. Platelet-bound antibody concentration increased with storage of anticoagulated blood prior to platelet isolation and with use of Na3 citrate and K3 EDTA anticoagulants. The preferred anticoagulant for platelet-bound antibody measurement is acid citrate dextrose. Platelet-bound antibody concentration should be determined not longer than 24 hours after blood collection.

The usefulness of tidal breathing flow-volume loops (TBFVL) to evaluate pulmonary function was investigated in 6 Standardbreds during treadmill exercise. Tidal breathing flow-volume loops are a graphic representation of airflow rate vs tidal volume for each individual breath. These TBFVL were obtained from horses exercising it speeds corresponding to 75 and 100% of maximum heart rate. Measurements were recorded in each horse before and after ovalbumininduced allergic lung disease. Moderate obstructive lung disease, characterized by a significant increase in pulmonary resistance, was observed while the horses were at rest. We found that in horses with airway obstruction exercising at 75 or 100% of maximum heart rate, the quantitative indices describing TBFVL shape and size were not markedly different from those in clinically normal horses exercising at similar speeds.

Schirmer tear test (STT), intraocular pressure (IOP) measurement, slit-lamp biomicroscopy, and indirect ophthalmoscopy were performed on 8 dogs with (13)l-induced hypothyroidism and 4 euthyroid control dogs at weeks 0, 9, 13, 17, immediately prior to treatment with levothyroxine, after 5 weeks of levothyroxine administration (0.022 mg/kg of body weight, PO, q 12 h), and at euthanasia 7 weeks after discontinuation of replacement therapy. Although the control group had higher baseline STT values than the hypothyroid group after randomization of dogs into the 2 groups (P < 0.01), STT values remained unchanged from their respective baseline values at all time intervals for both groups. Hypothyroid and control dogs had significant (P < 0.05) reduction in IOP from baseline values at all subsequent time points, but differences were not observed when hypothroid dogs were compared with controls. Goblet cell indices determined from biopsy samples of the inferior-nasal conjunctival fornix obtained before induction of hypothyroidism (baseline), immediately prior to and at conclusion of levothyroxine therapy, and at euthanasia were not significantly different when values for hypothyroid dogs were compared with their own baseline values or with values for control dogs. Histologic examination of the globes and adnexa at euthanasia also failed to indicate consistent qualitative differences between hypothyroid and control dogs. Marked reduction in serum thyroid hormone concentrations had little effect on the eye and ocular adnexa over the course of the study.

Seven adult mares were used to determine the analgesic, CNS, and cardiopulmonary effects of detomidine hydrochloride solution after epidural or subarachnoid administration, using both regimens in random sequence. At least 1 week elapsed between experiments. A 17-gauge Huber point (Tuohy) directional needle was used to place a catheter with stylet into either the epidural space at the first coccygeal interspace or the subarachnoid space at the lumbosacral intervertebral junction. Catheters were advanced so that the tips lay at the caudal sacral (S5 to S4) epidural space or at the midsacral (S3 to S2) subarachnoid space. Position of the catheter was confirmed radiographically. A 1% solution of detomidine HCl was injected into the epidural catheter at a dosage of 60 micrograms/kg of body weight, and was expanded to a 10-ml volume with sterile water to induce selective caudal epidural analgesia (CEA). A dose of 30 micrograms of detomidine HCl/kg expanded to a 3-ml volume with spinal fluid was injected into the subarachnoid catheter to induce caudal subarachnoid analgesia (CSA). Analgesia was determined by lack of sensory perception to electrical stimulation (avoidance threshold > 40 V, 0.5-ms duration) at the perineal dermatomes and no response to superficial and deep muscular pinprick stimulation at the pelvic limb and lumbar and thoracic dermatomes. Maximal CEA and CSA extended from the coccyx to spinal cord segments T15 and T14 at 10 to 25 minutes after epidural and subarachnoid drug administrations in 2 mares. Analgesia at the perineal area lasted longer after epidural than after subarachnoid administration (142.8 +/- 28.8 minutes vs 127.1 +/- 27.7 minutes). All mares remained standing. Both CEA and CSA induced marked sedation, moderate ataxia, minimal cardiopulmonary depression, increased frequency of second-degree atrioventricular heart block, and renal diuresis. All treatments resulted in significantly (P < 0.05) decreased heart rate, respiratory rate, systemic arterial blood pressure, PCV, and plasma total solids concentration. To the contrary, arterial carbon dioxide tension, plasma bicarbonate, and standard base excess concentrations were significantly (P < 0.05) increased. Arterial oxygen tension, pH, and rectal temperature did not change significantly from baseline values. Results indicate that use of detomidine for CEA and CSA in mares probably induces local spinal and CNS effects, marked sedation, moderate ataxia, mild cardiopulmonary depression, and renal diuresis.

We evaluated the feasibility of using miniosmotic pumps as a way to continuously treat cattle with a singular ergot alkaloid (ergonovine) of known content, thus mimicking the natural fescue toxicosis disease state, but allowing study of specific alkaloid effects. Dosing animals with increasing amounts of ergonovine via miniosmotic pumps, followed by daily acquisition of plasma samples for high-performance liquid chromatographic determination of the alkaloid, resulted in stepwise increases in plasma ergonovine concentration. However, despite the detectable blood concentration of ergonovine, calves did not have typical clinical signs of ergot alkaloid toxicosis. Similarly, serum prolactin concentration was unaffected by ergonovine in these cattle, implicating some other alkaloid of endophyte-infested fescue as causative of the usual prolactin-suppressive response. The results confirm use of this animal dosing method to study biological effects of singular purified alkaloids of known amount, without bioavailability concerns. Thus, this dosing method will facilitate studies to determine the harmful effects of individual alkaloids found in toxic tall fescue, and ultimately, to alleviate their costly effects in cattle, horses, and other species.

Three methods of determining glomerular filtration rate (GFR) were performed in adult ferrets, 9 months to 7 years old. Endogenous creatinine clearance was determined, using serum and urine creatinine values obtained during 24- and 48-hour collection periods from 27 ferrets housed in metabolic cages. Creatinine and radiolabeled inulin were administered to 12 female ferrets by constant IV infusion during isoflurane-induced anesthesia. Serial 20-minute urine collections, together with serum samples obtained at the midpoint of urine collection, provided measures for clearance calculations of these substances. Mean +/- SD endogenous creatinine clearance in ferrets for metabolic cage collections was 2.50 +/- 0.93 ml/min/ kg of body weight. There were no significant differences between the 24- and 48-hour clearance rates. Mean inulin clearance was 3.02 +/- 1.78, and mean exogenous creatinine clearance was 3.32 +/- 2.16 ml/ min/kg. Analysis of variance, using least-squared means adjustment, did not yield any significant differences between inulin and exogenous creatinine clearance rates. Exogenous creatinine clearance-to-inulin clearance ratio was 0.99 +/- 0.46, and there was significant correlation between the 2 methods (r = 0.82, P = 0.0001). Significant body temperature effects on inulin or exogenous creatinine clearance were not found. Infused inulin clearance, the generally preferred method for GFR calculation in mammalian species, was significantly (P = 0.0069) higher in younger (3.65 ml/min/kg) vs older ferrets (2.29 ml/min/kg). Results of this study indicate that inulin clearance is an adequate measure of GFR in ferrets as it is in other species. Compared with inulin clearance, exogenous creatinine clearance also provides a reliable estimate of GFR in ferrets.

A total of 21 calves were inoculated IV with 1 of the following isolates of bovine viral diarrhea virus (BVDV); CD87 (n = 10), NY-1 (n = 3), NADL (n = 5), or were sham-inoculated with virus-free medium (n = 3). Subsequent to inoculation, platelet counts were monitored to detect differences between noncytopathic (CD87, NY-1) and cytopathic (NADL) isolates in their ability to induce thrombocytopenia. Platelet count decrease throughout infection was statistically analyzed by comparing the slope of the line drawn from the count on the day of infection to the lowest count achieved by that calf. Significant difference was observed in the CD87-inoculated calves and in the NY-1-inoculated calves, compared with those of the same control group. Significant difference was not observed in the slope of platelet count decrease between the cytopathic NADL-infected calves and control-group calves. The data indicate that noncytopathic BVDV isolates may more easily induce thrombocytopenia than do cytopathic isolates in immune-naive, immunocompetent calves; acute infection with 1 cytopathic BVDV isolate (NADL) did not induce thrombocytopenia. In addition, although each calf seroconverted, virus was rarely isolated from mononuclear cells obtained from calves with cytopathic infections. At some point after infection, virus was always isolated from each of the calves undergoing noncytopathic infections, and occasionally, transient association of noncytopathic BVDV antigen with platelets was observed during these infections.