Quantification of presumptive Na+/H+ antiporters of the erythrocytes of trout and eel

The presumptive Na⁺/H⁺ exchange sites of trout and eel erythrocytes were quantified using amiloride-displaceable 5-(N-methyl-N-[³H]isobutyl)-amiloride (³H-MIA) equilibrium binding to further evaluate the mechanisms of i) hypoxia-mediated modifications in the trout erythrocyte -adrenergic signal transduction system and ii) the marked differences in the catecholamine responsiveness of this system between the trout and eel. MIA was a more potent inhibitor of both trout apparent erythrocyte proton extrusion (IC₅₀ = 20.1 ± 1.1 mol l^–1, N = 6) activity (as evaluated by measuring plasma pH changes after addition of catecholamine in vitro) and specific ³H-MIA binding (IC₅₀ = 257 ± 8.2 nmol l^–1, N = 3) than amiloride, which possessed a proton extrusion IC₅₀ of 26.1 ± 1.6 mol l^–1 (N = 6) and a binding IC₅₀ of 891 ± 113 nmol l^–1 (N = 3). The specific Na⁺ channel blocker phenamil was without effect on adrenergic proton extrusion activity or specific ³H-MIA binding. Trout erythrocytes suspended in Na⁺-free saline and maintained under normoxic conditions possessed 37,675 ± 6,678 (N = 6) amiloride-displaceable ³H-MIA binding sites per cell (B_max, presumptive Na⁺/H⁺ antiporters) with an apparent dissociation constant (K_D) of 244 ± 29 nmol l^–1 (N = 6). Acute hypoxia (PO₂ = 1.2 kPa; 30 min) did not affect the K_D, yet resulted in a 65% increase in the number of presumptive Na⁺/H⁺ antiporters. Normoxic eel erythrocytes, similarly suspended in Na⁺-free saline, possessed only 17,133 ± 3,716 presumptive Na⁺/H⁺ antiporters (N = 6), 45% of that of trout erythrocytes, with a similar K_D (246 ± 41 nmol l^–1, N = 6). These findings suggest that inter- and intra-specific differences in the responsiveness of the teleost erythrocyte -adrenergic signal transduction system can be explained, in part, by differences in the numbers of Na⁺/H⁺ exchange sites.     

Clinicopathologic features of suspected brevetoxicosis in double-crested cormorants (Phalacrocorax auritus) along the Florida Gulf Coast.

Outbreaks of morbidity and mortality in double-crested cormorants (Phalacrocorax auritus) along Florida's Gulf Coast have occurred sporadically for at least 30 yr. During these outbreaks, the Clinic for the Rehabilitation of Wildlife, located on Sanibel Island in Florida, has admitted a substantial number of cormorants with consistent presentation of primarily neurologic clinical signs. In order to investigate the association of these outbreaks in cormorants with exposure to brevetoxin, we compared the timing of admittance of cormorants with outbreak-specific clinical signs to blooms of the brevetoxin-producing marine algae, Karenia brevis (formerly Gymnodinium breve), around Sanibel Island from 1995 through 1999. The clinic admitted 360 out of 613 cormorants with the common clinical sign of severe cerebellar ataxia in six outbreaks occurring during this period. The ataxia was characterized by a broad-based stance, truncal incoordination, hypermetric gait, and intention tremors of the head. The histopathologic findings in 10 cormorants euthanized in 1997 were mild and nonspecific. An immunohistochemical staining technique for the detection of brevetoxin in cormorants documented the uptake of brevetoxin in tissues from four cormorants admitted during an outbreak in 1997, but a modified technique used on samples from 11 cormorants admitted during a K. brevis bloom in 2000 produced indeterminate results. Admittance of cormorants with outbreak-specific clinical signs was positively correlated (P < 0.05) with concurrent concentrations of K. brevis in local water. The cross-correlation coefficient was also significant when increased K. brevis levels preceded cormorant admittances by 2, 4, 6, and 8 wk. This delay in time between K. brevis blooms and cormorant admittance and our clinical finding of neurologic abnormalities in cormorants without overt histopathologic features suggest an association between K. brevis blooms and local cormorant morbidity.     

Presumptive fenbendazole toxicosis in North American porcupines

CASE DESCRIPTION: 4 North American porcupines were evaluated because of diarrhea or neutropenia (or both) that developed after treatment with fenbendazole for intestinal parasites. CLINICAL FINDINGS: Complete blood cell count abnormalities included severe neutropenia in all affected porcupines and mild anemia in some of them. In 2 porcupines, postmortem findings included bone marrow hypoplasia and intestinal crypt cell necrosis. TREATMENT AND OUTCOME: Affected porcupines received supportive care including fluid supplementation and broad-spectrum antimicrobials. The 2 surviving animals recovered after 9 to 33 days of treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Fenbendazole is an anthelminthic that may be used in an extralabel manner for the treatment of intestinal parasitism in wildlife species. The drug inhibits mitosis and can affect rapidly dividing cell lines, such as those in the bone marrow and intestinal crypt mucosa. Fenbendazole may not be an appropriate anthelminthic choice in North American porcupines.     

Equine Granulocytic Anaplasmosis: A Case Report and Review

A 15 year-old Thoroughbred mare was examined for lethargy, fever, and inappetence of 1-day duration. A hard-bodied tick was removed from the horse. A complete blood count (CBC) demonstrated leukopenia with lymphopenia and thrombocytopenia. Morulae were visualized in circulating granulocytes. A polymerase chain reaction (PCR) confirmed the identity of these organisms as Anaplasma phagocytophilum. The horse was treated symptomatically for fever and inappetence with flunixin meglumine (1.1 mg/kg [0.5 mg/lb]) and oral electrolyte paste. Oxytetracycline (6.6 mg/kg [3 mg/lb] intravenously every 24 hours) treatment was begun as soon as a definitive diagnosis was determined. The mare responded to treatment, but she was switched to oral doxycycline (10 mg/kg [4.5 mg/lb] every 12 hours) after 5 days because of perivascular swelling at the injection site. Complete resolution of clinical signs was seen. There was no evidence of recurrence 1 year later. No additional horses at the farm were affected. The horse in this report presented for lethargy, inappetence, and fever, with limited other abnormalities. This represents a classical presentation of a mild to moderate case of anaplasmosis, which had not previously been reported in Virginia. The disease may be more widespread than has been previously reported, and it should warrant inclusion on a complete differential diagnosis list in a case of fever of unknown origin.     

Hemipristicola gunterae gen. n., sp. n. (Cestoda: Tetraphyllidea: Phyllobothriidae) from the snaggletooth shark, Hemipristis elongata (Carcharhiniformes: Hemigaleidae), from Moreton Bay, Australia

Helminthological examination of the snaggletooth shark, Hemipristis elongata (Klunzinger) (Carcharhiniformes: Hemigaleidae), from Moreton Bay, Queensland, Australia, yielded a phyllobothriid genus and species previously unknown to science. Hemipristicola gunterae gen. n., sp. n. is described here, and is placed in the subfamily Phyllobothriinae Braun, 1900. Of the other phyllobothriid genera, the new genus most closely resembles Paraorygmatobothrium in that both genera possess bothridia with a single loculus and apical sucker, post-vaginal testes and lateral vitellarium. Hemipristicola, however, differs from Paraorygmatobothrium in the morphology of the proximal bothridial surface microthrix, possessing serrate gladiate spinitriches with marginal serrations restricted to the distal half of the blade, and in the possession of a more extensive uterus, extending anteriorly from the anterior margin of the ovary to well past the level of the cirrus-sac. The new genus also differs from Paraorygmatobothrium by possessing testes that are more than one layer deep. Hemipristicola gunterae further differs from Paraorygmatobothrium species found in hemigaleid sharks in possessing vitelline follicles arranged in two lateral bands that are restricted to the lateral margins of the proglottid and not possessing a cephalic peduncle. Bayesian inference analysis of partial 28S rDNA data shows that H. gunterae forms a sister taxon to species of Paraorygmatobothrium. These two genera were resolved with high posterior probability support in the analysis. Hemipristicola gunterae is only the second phyllobothriid species to be described from Hemipristis elongata from Australian waters, and the fourth from the Australian hemigaleids.     

The effects of yohimbine on the pharmacokinetic parameters of detomidine in the horse

ObjectiveTo describe the pharmacokinetics of detomidine and yohimbine when administered in combination.Study designRandomized crossover design.AnimalsNine healthy adult horses aged 9 ± 4 years and weighing of 561 ± 56 kg.MethodsThree dose regimens were employed in the current study. 1) 0.03 mg kg^?1 detomidine IV (D), 2) 0.2 mg kg^?1 yohimbine IV (Y) and 3) 0.03 mg kg^?1 detomidine IV followed 15 minutes later by 0.2 mg kg^?1 yohimbine IV (DY). Each horse received all three dose regimens with a minimum of 1 week in between subsequent regimens. Blood samples were obtained and plasma analyzed for detomidine and yohimbine concentrations by liquid chromatography-mass spectrometry. Data were analyzed using both non-compartmental and compartmental analysis.ResultsThe maximum measured detomidine concentrations were 76.0 and 129.9 ng mL^?1 for the D and DY treatments, respectively. Systemic clearance and volume of distribution of detomidine were not significantly different for either treatment. There was a significant increase in the maximum measured yohimbine plasma concentrations from Y (173.9 ng mL^?1) to DY (289.8 ng mL^?1). Both the Cl and V_d for yohimbine were significantly less (6.8 mL minute^?1 kg^?1 (Cl) and 1.7 L kg^?1 (V_d)) for the DY as compared to the Y treatments (13.9 mL minute^?1 kg^?1 (Cl) and 2.7 L kg^?1 (V_d)). Plasma concentrations were below the limit of quantitation (0.05 and 0.5 ng mL^?1) by 18 hours for both detomidine and yohimbine.Conclusion and clinical relevanceThe Cl and V_d of yohimbine were affected by prior administration of detomidine. The elimination half life of yohimbine remained unaffected when administered subsequent to detomidine. However, the increased plasma concentrations in the presence of detomidine has the potential to cause untoward effects and therefore further studies to assess the physiologic effects of this combination of drugs are warranted.     

The kinetics-based enzyme-linked immunosorbent assay for coronavirus antibodies in cats: calibration to the indirect immunofluorescence assay and computerized standardization of results through normalization to control values.

The computer-assisted, kinetics-based enzyme-linked immunosorbent assay for coronavirus antibodies in cats was calibrated to the conventional indirect immunofluorescence assay by linear regression analysis and computerized interpolation (generation of "immunofluorescence assay-equivalent" titers). Procedures were developed for normalization and standardization of kinetics-based enzyme-linked immunosorbent assay results through incorporation of five different control sera of predetermined ("expected") titer in daily runs. When used with such sera and with computer assistance, the kinetics-based enzyme-linked immunosorbent assay minimized both within-run and between-run variability while allowing also for efficient data reduction and statistical analysis and reporting of results.