Multiresidue assay for benzimidazole anthelmintics by liquid chromatography and confirmation by gas chromatography/selected-ion monitoring electron impact mass spectrometry

A multiresidue method utilizing all-disposable labware has been developed for 8 benzimidazole anthelmintics from ovine, bovine, and swine muscle and liver tissues. After an initial extraction with ethyl acetate and subsequent evaporation, a 3-component extraction using hexane, ethanol, and 0.2N HCl was used for final cleanup. Clean extracts were produced for separation and determination by reverse-phase liquid chromatography at 298 nm, using methanol and aqueous buffer as mobile phase. A synthesized internal standard, 2-(n-butylmercapto)benzimidazole, was used for quantitation of all drugs. Results are included along with statistical information verifying the performance of the method. Spiked control tissues and incurred drug tissues were used for an intralaboratory study with a concentration range of 50-1470 ppb. A series of standard curves at 0, 50, 100, and 200 ppb were analyzed. Overall recovery at the 100 ppb level averaged 92% (CV 8%) in liver tissues, across all 3 species and 88% (CV 5%) in muscle tissues across all 3 species. Results were confirmed by gas chromatography/mass spectrometry with acid hydrolysis of the remaining extract in 2N HCl followed by re-extraction of the amine and derivatization to the tert-butyldimethylsilyl derivative. The anthelmintics were identified by gas chromatography/selected ion monitoring electron-impact mass spectrometry. Ion ratio measurements were taken and compared to standard material. CVs averaged 10% or less for all drugs tested.     

Coagulopathic Effects and Therapy of Brodifacoum Toxicosis in Dogs

The clinical signs and laboratory changes of brodifacoum (BDF) intoxicated dogs and their response to vitamin K1 treatment were examined. Brodifacoum, a second-generation anticoagulant rodenticide, was fed to four dogs for 3 consecutive days producing a cumulative dose of 1.1 mg BDF/kg body weight. Clinical observations of the animals were made daily throughout the study. Monitored laboratory parameters included: one-stage prothrombin time (OSPT), activated partial thromboplastin time (APTT), activated coagulation time (ACT), complete blood counts, thrombocyte counts, and serum chemistry values. Response to vitamin K1 therapy was evaluated clinically and by laboratory tests. Serum BDF concentrations were monitored. Inappetence and hemorrhagic tendencies were exhibited by day 5 postrodenticide exposure. One-stage prothrombin time, APTT, and ACT were 25% greater than time zero values at 24, 24, and 72 hours postdosing, respectively. All laboratory parameters returned to normal within 48 hours of initiating vitamin K1 therapy (0.83 mg/kg orally, TID for 5 days). Serum brodifacoum concentrations were highest (1065-1215 ng/mL) during the 3 days after BDF dosing and were detectable (3.0-7.5 ng/mL) until day 24 postexposure. A mean BDF elimination half-life of 6 +/- 4 days was observed.     

Estimation of the relative bioavailability of copper sources in chicks fed on conventional dietary amounts

1. Tissue accumulation of Cu from dietary additions of 0, 5, 10, or 20 mg/kg Cu as reagent grade Cu acetate and feed grade Cu carbonate was determined in day‐old chicks fed on conventional maize‐soyabean meal starter diets (5.41 mg/kg Cu as‐fed basis) for 3 weeks.

2. Average daily food intake, daily weight gain and food conversion were similar among treatments.

3. There were linear increases in plasma and liver Cu concentrations (P< 0.01) as dietary Cu increased.

4. Bioavailability of Cu as carbonate was 0.66 that of Cu in the acetate based on the multiple regression slope ratio of liver Cu concentration on added dietary Cu. Although responses for the two Cu sources did not differ significantly, the relative bioavailability of the Cu carbonate was similar (0.66 vs 0.68) to that obtained in an earlier study (Ledoux et al., 1991) with greater dietary Cu contents (150, 300 and 450 mg/kg) in which the slopes of the equations representing the two sources differed (P<0.05).     

Calving patterns in seasonal dairy herds

Two field studies examined the calving patterns of cows in seasonal dairy herds in the Waikato (Field Study 1) and South Taranaki regions (Field Study 2). The first study examined patterns for cows commencing their second or subsequent lactation in herds which had used an inseminating service during the previous season. The second study included first lactation heifers only in 15 herds where animals had been naturally mated, and in 15 herds in which they had been synchronised and then artificially inseminated at the synchronised oestrus. The parameters describing calving patterns were based on the date for each herd's planned start of calving (PSC), which was 282 days from the date on which breeding commenced in the preceding season. The average interval from PSC to mean calving date for the 35 herds in Field Study 1 was 22 days, with individual herds ranging from 15 to 30 days. In herds with heifers which had been naturally mated (Field Study 2), it was 17.6 days compared to 11.0 days for previously synchronised animals. Calculating the intervals from PSC to median calving date and separately for the last two quartiles more effectively described a herd's calving pattern. The duration for the last quartile of the calving pattern was influenced by the extent and timing of induced calving. In Field Study 1, 88.6% of the 35 herd owners induced premature parturition in at least one cow. In these herds, 11.3% of cows were treated and calved prematurely. Only 61.7% of heifers which had previously been naturally mated calved by 3 weeks after PSC. Their calving dates were not evenly distributed over this 3-week period, with 9.8% in the first week and 25.6% in the third week. The calving pattern for heifers which had been previously synchronised showed several distinct peaks. Calvings to the synchronised mating were completed 15 days after PSC, by which time 64.7% of animals had calved. By 3 weeks after PSC, 72.9% of these heifers had calved. The results showed that there was considerable variation in calving patterns in seasonal dairy herds. This variation would have been due to differences in conception pattern, and the way induced calving had been applied. The calving pattern in heifers which had been naturally mated was less concentrated than had been expected. Synchronisation can significantly concentrate the calving pattern of these first lactation animals. The parameters used to describe calving patterns may be less applicable in herds in which a high proportion of animals is induced to calve prematurely, or where a whole herd is synchronised. Nonetheless, they do serve as an illustrative example of the variation in calving patterns among herds.     

MAGNETIC RESONANCE IMAGING FINDINGS IN SPINAL CORD INFARCTION IN THREE SMALL BREED DOGS

Fibrocartilaginous embolization (FCE) of the spinal cord is a common disease in large breed dogs. There are only a few reports about this entity in small breed dogs and it has never been reported in chondrodystrophic breed. For definitive diagnosis histopathologic examination is necessary. Magnetic resonance imaging (MRI) as a potential diagnostic tool for intravitam diagnosis of FCE has been mentioned before, but results have not been reported so far. This report describes the neurological findings and MRI results in three small breed dogs, including a Pekingese dog, with FCE of the spinal cord. The disease was suspected in two animals based upon clinical and MRI-appearance and confirmed in the third by histopathological examination. In all three cases, similar focal intramedullary lesions, consisting of hyperintensive signals on T2-weighted images, were detected. Based on these findings, high-field MRI may be used as an antemortem tool for the diagnosis of FCE. It is also shown that FCE can occur in chondrodystrophic dogs.     

Carbon storage by urban tree cultivars, in roots and above-ground

Urban trees can favorably affect factors underlying global warming by storing carbon and by reducing energy needs for cooling and heating buildings. To estimate carbon stored in roots and above-ground portions of trees, data was collected consisting of whole tree sampling of Amelanchier, Malus, Pyrus, and Syringa cultivars. Roots were excavated using an Air-Spade^™. Regression analysis resulted in two equations for predicting total carbon storage based on height and diameter of trees up to 20 cm dbh: Y = 0.05836 (dbh²) for root carbon storage, and Y = 0.0305 (dbh² × h)^0.9499 for above-ground carbon storage, explaining 97% and 96% of the variation, respectively. Average carbon stored in roots of various cultivars ranged from 0.3 to 1.0 kg for smaller trees, those 3.8 to 6.4 cm dbh, to more than 10.4 kg for trees 14.0 cm to 19.7 cm dbh. Average total carbon stored by cultivars ranged from 1.7 to 3.6 kg for trees less than 6.4 cm dbh to 54.5 kg for trees larger than 14.0 cm. The data from these equations apply mainly to trees in nurseries and recently transplanted trees. Comparisons showed that above-ground estimates from previous studies using a sampling technique overestimated values obtained from actual above-ground weights.     

Clinical Evaluation of Tavocept to Decrease Diuresis Time and Volume in Dogs with Bladder Cancer Receiving Cisplatin

Background

Transitional cell carcinoma is the most common bladder cancer of dogs. Cisplatin combined with piroxicam provides superior response rates, but unacceptable rates of nephrotoxicity. Tavocept is a chemoprotectant that has mitigated cisplatin toxicity and decreased the required infusion/diuresis volume in clinical trials in humans.

Hypothesis/Objectives

We hypothesized that Tavocept would decrease diuresis volume and time and facilitate safe administration of a cisplatin/piroxicam protocol to dogs with bladder cancer. Secondary objectives were to compare response rate and survival times to an historical comparator group treated without Tavocept.

Animals

Fourteen client‐owned dogs were prospectively enrolled.

Methods

Tumor volume was measured by computed tomography at days 0, 42, and 84. Dogs received combination Tavocept/cisplatin with a shortened diuresis protocol. A total of 4 doses was planned, with concurrent administration of piroxicam. Serial biochemical analyses were evaluated for azotemia.

Results

A 90‐minute infusion/diuresis time was used for all dogs. Three dogs (21%) had concurrent increases in serum creatinine (>2.0 mg/dL) and BUN (>42 mg/dL) concentrations; 2 of these dogs were isosthenuric. This frequency of nephrotoxicity is significantly less (P = 0.0406) than that of an historical control group treated without Tavocept. Overall response rate was 27%. Median survival time was comparable to historical controls (253 vs. 246 days).

Conclusions and Clinical Importance

Tavocept decreased the required diuresis time with cisplatin from > 6 hours to 90 minutes, while also decreasing occurrence of azotemia. Survival time was comparable, but the response rate was inferior to an historical comparator group. Further evaluation in other tumors susceptible to platinum agents is warranted.