Bupivacaine in the horse: relationship of local anaesthetic responses and urinary concentrations of 3-hydroxybupivacaine     

J D Harkins  A Lehner  W Karpiesiuk  W E Woods  L Dirikolu  J Boyles  W G Carter  T Tobin 《Journal of veterinary pharmacology and therapeutics》1999,22(3):181-195

Bupivacaine is a potent local anaesthetic used in equine medicine. It is also classified as a Class 2 foreign substance by the Association of Racing Commissioners International (ARCI). The identification of residues in postrace urine samples may cause regulators to impose significant penalties. Therefore, an analytical/pharmacological database was developed for this medication. The highest no-effect dose (HNED) for the local anaesthetic effect of bupivacaine was determined to be 0.25 mg by using an abaxial sesamoid local anaesthetic model. Administration of the HNED of bupivacaine to eight horses yielded a peak urine concentration of apparent bupivacaine of 23.3 ng/mL 2 h after injection as determined with enzyme-linked immunosorbent assay (ELISA) screening. The major metabolite recovered from beta-glucuronidase-treated equine urine after dosing with bupivacaine is a hydroxybupivacaine, either 3-hydroxybupivacaine, 4-hydroxybupivacaine, or a mixture of the two. To determine which positional isomer occurs in the horse, 4-hydroxybupivacaine was obtained from Maxxam Analytics, Inc., and 3-hydroxybupivacaine was synthesized, purified, and characterized. Furthermore, a quantitative mass spectrometric method was developed for the metabolite as recovered from horse urine. Following subcutaneous injection of the HNED of bupivacaine, the concentration of the hydroxybupivacaine recovered from horse urine reached a peak of 27.4 ng/mL at 4 h after administration as measured by gas chromatography/mass spectrometry (GC/MS). It was also unequivocally demonstrated with ion chromatography that the hydroxybupivacaine metabolite found in horse urine is exclusively 3-hydroxybupivacaine and not 4-hydroxybupivacaine. The mean pH of the 4-h urine samples was 7.21; the mean urine creatinine was 209.5 mg/dL; and the mean urine specific gravity was 1.028. There was no apparent effect of pH, urine creatinine concentration, or specific gravity on the concentration of 3-hydroxybupivacaine recovered. The concentration of bupivacaine or its metabolites after administration of a HNED dose are detectable by mass spectrometric techniques. This study also suggests that recovery of concentrations less than approximately 30 ng/mL of 3-hydroxybupivacaine from postrace urine samples is unlikely to be associated with a recent local anaesthetic effect of bupivacaine.  相似文献   

Growth, Survival, and Body Composition of Cage-Cultured Nile Tilapia Oreochromis niloticus Fed Pelleted and Unpelleted Distillers Grains with Solubles in Polyculture with Freshwater Prawn Macrobrachium rosenbergii     

James H.  Tidwell  Shawn D.  Coyle  Aaron  VanArnum  Charles  Weibel Susan  Harkins 《Journal of the World Aquaculture Society》2000,31(4):627-631

Abstract.— A 12‐wk feeding trial was conducted in cages with juvenile Nile tilapia Oreochromis niloticus to evaluate distillers grains with solubles (DDGS) as a direct feed, the effects of pelleting on its utilization, and the compatibility of caged tilapia and prawns in polyculture. Nine 1.0‐m³ cages were stocked with 200 juvenile (26 ± 0.9 g) tilapia. Cages were suspended in a 0.2‐ha pond stocked with juvenile freshwater prawns Macrobrachium rosenbergii at 40,000/ha. Three replicate cages were randomly assigned to each dietary treatment. In one dietary treatment DDGS was fed as an unpelleted loose grain ration (26% protein). In a second dietary treatment fish were fed DDGS that had been steam‐pelleted (23% protein). Fish in a third dietary treatment were fed a commercial catfish diet (31% protein) for comparison. After 12 wk, individual weight, individual length, and specific growth rate were significantly higher (P < 0.05) and feed conversion ratio was significantly lower (P < 0.05) for fish fed the commercial catfish diet than for fish fed either unpelleted or pelleted DDGS. Specific growth rate was significantly higher (P < 0.05) for fish fed pelleted DDGS than for fish fed unpelleted DDGS. Survival did not differ significantly (P > 0.05) among treatments (>95%). Although growth was increased in fish fed the commercial diet, their cost of production (<0.66/kg gain) was significantly higher (P < 0.05) than in fish fed unpelleted and pelleted DDGS (<0.26/ kg gain and <0.37/kg gain, respectively). The costs of gain in fish fed unpelleted DDGS was significantly lower (P < 0.05) than in fish fed the pelleted DDGS. Prawn production was 1,449 kg/ha and addition of tilapia in polyculture increased total pond productivity approximately 81 %. These data suggest that DDGS provides economical growth in tilapia when fed as a direct feed and that polyculture of tilapia may improve overall pond efficiency in freshwater prawn production ponds, even at temperate latitudes.  相似文献   

Identification of hydroxyropivacaine glucuronide in equine urine by ESI+/MS/MS.          下载免费PDF全文

J D Harkins  W Karpiesiuk  T Tobin  L Dirikolu    A F Lehner 《Canadian journal of veterinary research》2000,64(3):178-183

Ropivacaine is a local anesthetic that has a high potential for abuse in racing horses. It can be recovered from urine collected after administration as a hydroxylated metabolite following beta-glucuronidase treatment of the urine. Based on these findings, it has been inferred that ropivacaine is present in equine urine as a glucuronide metabolite; however, these metabolites have never been directly identified. Using ESI+/MS/MS, the presence of a [M+H]+ molecular ion of m/z 467 was demonstrated in urine corresponding to the calculated mass of a hydroxyropivacaine glucuronide +1. The abundance of this ion diminished after glucuronidase treatment with concomitant appearance of a m/z 291 peak, which is consistent with its hydrolysis to hydroxyropivacaine. In further work, the m/z 467 material was fragmented in the MS/MS system, yielding fragments interpretable as hydroxyropivacaine glucuronide. These data are consistent with the presence of a hydroxyropivacaine glucuronide in equine urine and constitute the first direct demonstration of a specific glucuronide metabolite in equine urine.  相似文献   

Western blot analysis of the IgG response of sheep vaccinated with S48 Toxoplasma gondii (Toxovax)     

J.M. Wastling  D. Harkins  D. Buxton 《Research in veterinary science》1994,57(3):384

The IgG antibody responses of sheep vaccinated by the subcutaneous injection of live tachyzoites of ‘incomplete’ strain S48 toxoplasma (Toxovax) were analysed by Western blotting. Antibodies corresponding to a range of tachyzoite antigens (13 to 48 kD) were detected, but the response was dominated by antibody recognising a 30 to 32 kD band. Unvaccinated ewes challenged orally with oocysts of the ‘complete’ M3 toxoplasma strain had a more complex IgG response that recognised antigens in six dominant bands of similar intensity as those in sheep vaccinated with S48 tachyzoites and then challenged with M3 oocysts. No differences were detected between the antigenic structures of the S48 tachyzoites and RH strain tachyzoites when the antigens were probed with immune ovine sera. Many of the anitgens of the S48 tachyzoites that were recognised had molecular weights similar to those of antigens that have been identified in other strains of toxoplasma.  相似文献   

Diclazuril in the horse: its identification and detection and preliminary pharmacokinetics   总被引：1，自引：0，他引：1  

Dirikolu L  Lehner F  Nattrass C  Bentz BG  Woods WE  Carter WG  Karpiesiuk W  Jacobs J  Boyles J  Harkins JD  Granstrom DE  Tobin T 《Journal of veterinary pharmacology and therapeutics》1999,22(6):374-379

Diclazuril (4-chlorophenyl [2,6-dichloro-4-(4,5-dihydro-3H-3,5-dioxo-1,2,4-triazin-2-yl)pheny l] acetonitrile), is a benzeneacetonitrile antiprotozoal agent (Janssen Research Compound R 64433) marketed as Clinacox . Diclazuril may have clinical application in the treatment of Equine Protozoal Myeloencephalitis (EPM). To evaluate its bioavailability and preliminary pharmacokinetics in the horse we developed a sensitive quantitative high-pressure liquid chromatography (HPLC) method for diclazuril in equine biological fluids. MS/MS analysis of diclazuril in our HPLC solvent yielded mass spectral data consistent with the presence of diclazuril. After a single oral dose of diclazuril at 2.5 g/450 kg (as 500 g Clinacox), plasma samples from four horses showed good plasma concentrations of diclazuril which peaked at 1.077 +/- 0.174 microg/mL (mean +/- SEM) with an apparent plasma half-life of about 43 h. When this dose of Clinacox was administered daily for 21 days to two horses, mean steady state plasma concentrations of 7-9 microg/mL were attained. Steady-state levels in the CSF ranged between 100 and 250 ng/mL. There was no detectable parent diclazuril in the urine samples of dosed horses by HPLC or by routine postrace thin layer chromatography (TLC). These results show that diclazuril is absorbed after oral administration and attains steady-state concentrations in plasma and CSF. The steady state concentrations attained in CSF are more than sufficient to interfere with Sarcocystis neurona, whose proliferation is reportedly 95% inhibited by concentrations of diclazuril as low as 1 ng/mL. These results are therefore entirely consistent with and support the reported clinical efficacy of diclazuril in the treatment of clinical cases of EPM.  相似文献   

Flow cytometric analysis of canine colonic mucosal lymphocytes from endoscopically obtained biopsy specimens     

Sonea IM  Harkins K  Wannemuehler MJ  Jergens AE  Merten EA  Sacco RE  Cunnick JE 《American journal of veterinary research》1999,60(3):346-353

OBJECTIVE: To validate use of canine colonic biopsy specimens obtained via endoscopy as a source of mucosal lymphocytes (ML) for flow cytometric analysis. SAMPLE POPULATION: Mucosal biopsy specimens from 10 adult dogs. PROCEDURE: Mucosal lymphocyte subsets obtained from excised colon were compared with ML subsets obtained from biopsy specimens obtained by use of an endoscopic forceps (6 dogs). Endoscopic colonic biopsy specimens from 4 other dogs were used to define whether obtained ML were predominantly of intraepithelial or lamina propria origin. Mucosal lymphocytes were isolated and labeled, using commercially available monoclonal antibodies directed against canine cell surface antigens. Lymphocyte subsets (cytotoxic or helper T cells; B cells) were determined by use of flow cytometric analysis. RESULTS: A large number of viable ML was obtained after dissociation of the colonic epithelium from excised colon (45.5 + 21.5 X 10(6)) and endoscopic (7.2+/-3.4 X 10(6)) biopsy specimens. Lymphocyte subsets obtained with both methods were identical for each dog and consisted predominantly of intraepithelial lymphocytes, with some lymphocytes from the lamina propria. Collagenase digestion of excised colon also yielded a large number of viable lymphocytes from the lamina propria (56.7+/-20.4 X 10(6)), but collagenase digestion of endoscopic biopsy specimens was less rewarding. CONCLUSION AND CLINICAL RELEVANCE: A representative sample of viable intraepithelial ML is obtainable from endoscopic biopsy specimens. Flow cytometric analysis, a minimally invasive technique, can be used to study ML of client-owned animals.  相似文献   

The detection and biotransformation of guanabenz in horses: a preliminary report     

Harkins JD  Dirikolu L  Lehner AF  Hughes C  Schroedter D  Mayer B  Bratton C  Fisher MV  Tobin T 《Veterinary therapeutics : research in applied veterinary medicine》2003,4(2):197-209

Guanabenz (2,6-dichlorobenzylidene-amino-guanidine) is a centrally acting antihypertensive drug whose mechanism of action is via alpha2 adrenoceptors or, more likely, imidazoline receptors. Guanabenz is marketed as an antihypertensive agent in human medicine (Wytensin tablets, Wyeth Pharmaceuticals). Guanabenz has reportedly been administered to racing horses and is classified by the Association of Racing Commissioners International as a class 3 foreign substance. As such, its identification in a postrace sample may result in significant sanctions against the trainer of the horse. The present study examined liquid chromatographic/tandem quadrupole mass spectrometric (LC-MS/MS) detection of guanabenz in serum samples from horses treated with guanabenz by rapid i.v. injection at 0.04 and 0.2 mg/kg. Using a method adapted from previous work with clenbuterol, the parent compound was detected in serum with an apparent limit of detection of approximately 0.03 ng/ml and the limit of quantitation was 0.2 ng/ml. Serum concentrations of guanabenz peaked at approximately 100 ng/ml after the 0.2 mg/kg dose, and the parent compound was detected for up to 8 hours after the 0.04 mg/kg dose. Urine samples tested after administration of guanabenz at these dosages yielded evidence of at least one glucuronide metabolite, with the glucuronide ring apparently linked to a ring hydroxyl group or a guanidinium hydroxylamine. The LC-MS/MS results presented here form the basis of a confirmatory test for guanabenz in racing horses.  相似文献   

The mare reproductive loss syndrome and the eastern tent caterpillar: a toxicokinetic/statistical analysis with clinical, epidemiologic, and mechanistic implications     

Sebastian M  Gantz MG  Tobin T  Harkins JD  Bosken JM  Hughes C  Harrison LR  Bernard WV  Richter DL  Fitzgerald TD 《Veterinary therapeutics : research in applied veterinary medicine》2003,4(4):324-339

During 2001, central Kentucky experienced acute transient epidemics of early and late fetal losses, pericarditis, and unilateral endophthalmitis, collectively referred to as mare reproductive loss syndrome (MRLS). A toxicokinetic/statistical analysis of experimental and field MRLS data was conducted using accelerated failure time (AFT) analysis of abortions following administration of Eastern tent caterpillars (ETCs; 100 or 50 g/day or 100 g of irradiated caterpillars/day) to late-term pregnant mares. In addition, 2001 late-term fetal loss field data were used in the analysis. Experimental data were fitted by AFT analysis at a high (P <.0001) significance. Times to first abortion ("lag time") and abortion rates were dose dependent. Lag times decreased and abortion rates increased exponentially with dose. Calculated dose x response data curves allow interpretation of abortion data in terms of "intubated ETC equivalents." Analysis suggested that field exposure to ETCs in 2001 in central Kentucky commenced on approximately April 27, was initially equivalent to approximately 5 g of intubated ETCs/day, and increased to approximately 30 g/day at the outbreak peak. This analysis accounts for many aspects of the epidemiology, clinical presentations, and manifestations of MRLS. It allows quantitative interpretation of experimental and field MRLS data and has implications for the basic mechanisms underlying MRLS. The results support suggestions that MRLS is caused by exposure to or ingestion of ETCs. The results also show that high levels of ETC exposure produce intense, focused outbreaks of MRLS, closely linked in time and place to dispersing ETCs, as occurred in central Kentucky in 2001. With less intense exposure, lag time is longer and abortions tend to spread out over time and may occur out of phase with ETC exposure, obscuring both diagnosis of this syndrome and the role of the caterpillars.  相似文献   

Effects of induced alkalosis on performance in thoroughbreds during a 1,600-m race.     

J D Harkins  S G Kamerling 《Equine veterinary journal》1992,24(2):94-98

There is considerable debate regarding the ergogenic effects of sodium bicarbonate (NaHCO3) on racing performance in horses. Anecdotal evidence suggests that NaHCO3 improves performance by increasing the buffering capacity of the blood and delaying the onset of hydrogen ion-induced fatigue. In a cross-over study, 16 Thoroughbred racehorses were given an aqueous solution of NaHCO3 (0.4 g/kg in 1 litre H2O) or a control treatment (1 litre H2O) before a 1600-m race. Treatments were administered 3 h before the race, which was the time to peak buffering capacity (2.5-3.0 h) determined in a separate study. Before the race, there was a significant increase in venous HCO3- and pH in the NaHCO3-treated horses. After the race, there was a significant increase in venous blood pH and lactate in the NaHCO3-treated horses. Collectively, the data suggest an improved buffering capacity of the blood after NaHCO3 treatment. However, there was no change in race times or venous partial pressure of carbon dioxide. Therefore, the administration of NaHCO3 provided no ergogenic benefit to horses competing in a 1,600-m race.  相似文献   

Effect of recombinant porcine somatotropin on lactational performance and metabolite patterns in sows and growth of nursing pigs   总被引：1，自引：0，他引：1  

M Harkins  R D Boyd  D E Bauman 《Journal of animal science》1989,67(8):1997-2008

Sixteen crossbred sows (Yorkshire x Duroc) were used to determine the effect of recombinantly derived porcine somatotropin (pST) on lactational performance and the pattern of plasma metabolites and growth rate of nursing pigs. Daily s.c. injections of either pST (8.22 mg.sow-1.d-1) or excipient were administered at 1000 on d 12 through d 29 of lactation. Jugular cannulas were inserted in three sows/treatment and hourly blood samples were collected on d 11 to 13 and d 28 to 29 of lactation to determine the effect of treatment on plasma concentrations of somatotropin, glucose and nonesterified fatty acids in plasma. Milk production and weight of nursing pigs were determined pretreatment (d 9 and 10) and on d 16, 22 and 28. Milk production of sows receiving pST progressively increased above that of control sows and was 22% greater on d 28. Milk composition was not affected by treatment with pST (P greater than .10), so that the increase in yields of milk fat, lactose and solids paralleled the increases in milk yield. Total milk protein yield tended to be higher in sows receiving pST, but protein yield was greater (P less than .10) only on d 28. Pigs suckling sows treated with pST weighed .34 kg more at the end of the lactation period (P less than .05). Sows receiving pST consumed less feed (P less than .05) during the treatment period, and, as a result, lost more weight (P less than .10) and backfat (P less than .05) than control sows. Average concentrations of plasma somatotropin were elevated approximately 2.5-fold above baseline levels by exogenous pST. No acute alterations in plasma glucose or nonesterified fatty acids were observed in response to pST treatment, however, sows receiving pST had a chronic elevation of plasma glucose on d 29 of lactation.  相似文献