Pharmacologic effects and detection methods of methylated analogs of fentanyl in horses

Pharmacologic effects of alpha-methylfentanyl and 3-methylfentanyl, analogs of fentanyl, were investigated in mares. The ability of an 125I-labeled fentanyl radioimmunoassay (125I-RIA) to detect these methylated fentanyl analogs in individual and pooled urine samples from horses was evaluated. Also, the ability of 7 fentanyl antibodies to react with fentanyl and fentanyl derivatives (sufentanil, alfentanil, and carfentanil) was investigated. Mares were studied in a locomotor test to determine the amount of stimulation methylated fentanyl analogs might induce. Two mares each were given alpha-methylfentanyl at 1, 2, 4, 8, or 13 micrograms/kg of body weight, IV, or 3-methylfentanyl at 0.4, 0.7, or 1 microgram/kg IV. The cross-reactivity of sufentanil, alfentanil, carfentanil, alpha-methylfentanyl, and 3-methylfentanyl with 7 fentanyl antibodies was studied, using the 125I-RIA. All fentanyl analogs, with the exception of alfentanil, cross-reacted well with a C1 antibody raised to fentanyl. Less satisfactory cross-reactivity was determined with 6 other antibodies raised to fentanyl derivatives. When the C1 antibody was combined with an iodinated analog to fentanyl, good detectability of alpha-methylfentanyl and 3-methylfentanyl, in terms of fentanyl equivalents, was obtained from urine samples of dosed mares. The ability of the 125I-RIA to detect methylated fentanyl analogs in forensic urine samples pooled in groups of up to 20 samples was evaluated. When these methylated analogs were administered to mares in doses that induced measurable locomotor stimulation, the analog's presence was readily detected in individual or pooled samples.     

Pheromone orientation: role of internal control mechanisms

Male American cockroaches walk a zigzag path upwind toward a source of female sex pheromone. Although the maximum width of the pathway is regulated by the width of an odor plume, many turns are made before the edge of a wide plume is encountered. In addition to the pheromone regulation of the insect's orientation movements, an internal mechanism appears to influence the zigzag turning pattern.     

Detection, quantification, and pharmacokinetics of furosemide and its effects on urinary specific gravity following IV administration to horses

Furosemide is a potent loop diuretic used for the prevention of exercise-induced pulmonary hemorrhage in horses. This drug may interfere with the detection of other substances by reducing urinary concentrations, so its use is strictly regulated. The regulation of furosemide in many racing jurisdictions is based on paired limits of urinary SG (<1.010) and serum furosemide concentrations (>100 ng/ml). To validate this regulatory mechanism, a liquid chromatography/mass spectrometry/mass spectrometry method employing a solid-phase extraction procedure and furosemide-d5 as an internal standard was developed. The method was used to determine the pharmacokinetic parameters of furosemide in equine serum samples and its effects on urinary SG after IV administration (250 mg) to 10 horses. Pharmacokinetic analysis showed that serum concentrations of furosemide were well described by a two-compartmental open model. Based on results in this study, it is very unlikely for horses to have serum furosemide concentrations greater than 100 ng/ml or urine SG less than 1.010 at 4 hours after administration (250 mg IV). However, it should be remembered that urine SG is a highly variable measurement in horses, and even without furosemide administration, some horses might naturally have urine SG values less than 1.010.     

Effect of furosemide on urine specific gravity and osmolality in thoroughbred racehorses

Postrace urine samples from thoroughbred horses were examined to compare osmolality and specific gravity between horses treated with furosemide and those not treated. Samples were assigned to groups in relation to reported medication (furosemide) status, race finish position, and distance of race. Urine osmolality was significantly (P <.05) lower in samples from horses treated with furosemide when compared with untreated horses. Specific gravity determinations are less precise at measuring urine osmolality at lower levels (1.01 g/ml or less). The measurement of osmolality is a superior method for determining the urine solute concentration and facilitating the regulation of furosemide.     

Lidocaine in the horse: its pharmacological effects and their relationship to analytical findings

Lidocaine is a local anaesthetic agent that is widely used in equine medicine. It is also an Association of Racing Commissioners International (ARCI) Class 2 foreign substance that may cause regulators to impose substantial penalties if residues are identified in post race urine samples. Therefore, an analytical/pharmacological database was developed for this drug. Using our abaxial sesamoid local anaesthetic model, the highest no-effect dose (HNED) for the local anaesthetic effect of lidocaine was determined to be 4 mg. Using enzyme-linked immunosorbent assay (ELISA) screening, administration of the HNED of lidocaine to eight horses yielded peak serum and urine concentrations of apparent lidocaine of 0.84 ng/mL at 30 min and 72.8 ng/mL at 60 min after injection, respectively. These concentrations of apparent lidocaine are readily detectable by routine ELISA screening tests (LIDOCAINE ELISA, Neogen, Lexington, KY). ELISA screening does not specifically identify lidocaine or its metabolites, which include 3-hydroxylidocaine, dimethylaniline, 4-hydroxydimethylaniline, monoethylglycinexylidine, 3-hydroxymonoethylglycinexylidine, and glycinexylidine. As 3-hydroxylidocaine is the major metabolite recovered from equine urine, it was synthesized, purified and characterized, and a quantitative mass spectrometric method was developed for 3-hydroxylidocaine as recovered from horse urine. Following subcutaneous (s.c.) injection of the HNED of lidocaine, the concentration of 3-hydroxylidocaine recovered from urine reached a peak of about 315 ng/mL at 1 h after administration. The mean pH of the 1 h post dosing urine samples was 7.7, and there was no apparent effect of pH on the amount of 3-hydroxylidocaine recovered. Within the context of these experiments, the data suggests that recovery of less than 315 ng/mL of 3-hydroxylidocaine from a post race urine sample is unlikely to be associated with a recent local anaesthetic effect of lidocaine. Therefore these data may be of assistance to industry professionals in evaluating the significance of small concentrations of lidocaine or its metabolites in postrace urine samples. It should be noted that the quantitative data are based on analytical methods developed specifically for this study, and that methods used by other laboratories may yield different recoveries of urine 3-hydroxylidocaine.     

Above- and belowground biomass measurements in an unthinned stand of Sitka spruce (<Emphasis Type="Italic">Picea sitchensis</Emphasis> (Bong) Carr.)

Reporting carbon (C) stocks in tree biomass (above- and belowground) to the United Nations Framework Convention on Climate Change (UNFCCC) should be transparent and verifiable. The development of nationally specific data is considered ‘good practice’ to assist in meeting these reporting requirements. From this study, biomass functions were developed for estimating above- and belowground C stock in a 19-year-old stand of Sitka spruce (Picea sitchensis (Bong) Carr.). Our estimates were then tested against current default values used for reporting in Ireland and literature equations. Ten trees were destructively sampled to develop aboveground and tree component biomass equations. The roots were excavated and a root:shoot (R) ratio developed to estimate belowground biomass. Application of the total aboveground biomass function yielded a C stock estimate for the stand of 74 tonnes C ha⁻¹, with an uncertainty of 7%. The R ratio was determined to be 0.23, with an uncertainty of 10%. The C stock estimate of the belowground biomass component was then calculated to be 17 tonnes C ha⁻¹, with an uncertainty of 12%. The equivalent C stock estimate from the biomass expansion factor (BEF) method, applying Ireland’s currently reported default values for BEF (inclusive of belowground biomass), wood density and C concentration and methods for estimating volume, was found to be 60 tonnes C ha⁻¹, with an uncertainty of 26%. We found that volume tables, currently used for determining merchantable timber volume in Irish forestry conditions, underestimated volume since they did not extend to the yield of the forest under investigation. Mean stock values for belowground biomass compared well with that generated using published models.     

Assessment of allometric algorithms for estimating leaf biomass, leaf area index and litter fall in different-aged Sitka spruce forests

The relationship between leaf area and diameter at breast height(d.b.h.) or sapwood area (AS) has been used to estimate standleaf area or biomass of forest canopies. It has been suggestedthat intra-specific variations in the relationship between standleaf area and d.b.h. or AS can introduce a systematic errorin these estimates for younger and older stands unless additionalparameters relating to canopy structure are included in allometricfunctions. We collected data from a Sitka spruce chronosequenceto parametrize and test different algorithms for the estimationof foliar biomass (FB) and litter inputs over a range of forestages. FB estimates were significantly improved when additionalbiometric information relating to crown structure (canopy opennessand height of live crown) was included in the models. Althoughthe use of the relationship between leaf area and AS for theestimation of leaf area is justified by theoretical considerations(pipe model theory), we show that d.b.h. and other canopy parametersprovided the most robust estimation of leaf area across different-agedstands. Our results also suggest that the accuracy of litterinput estimates depends on needle retention time and annualturnover rate, particularly immediately before and after canopyclosure.     

Multiple‐Aetiology Enteric Infections Involving Non‐O157 Shiga Toxin‐Producing Escherichia coli – FoodNet, 2001–2010

We describe multiple‐aetiology infections involving non‐O157 Shiga toxin‐producing Escherichia coli (STEC) identified through laboratory‐based surveillance in nine FoodNet sites from 2001 to 2010. A multiple‐aetiology infection (MEI) was defined as isolation of non‐O157 STEC and laboratory evidence of any of the other nine pathogens under surveillance or isolation of >1 non‐O157 STEC serogroup from the same person within a 7‐day period. We compared exposures of patients with MEI during 2001–2010 with those of patients with single‐aetiology non‐O157 STEC infections (SEI) during 2008–2009 and with those of the FoodNet population from a survey conducted during 2006–2007. In total, 1870 non‐O157 STEC infections were reported; 68 (3.6%) were MEI; 60 included pathogens other than non‐O157 STEC; and eight involved >1 serogroup of non‐O157 STEC. Of the 68 MEI, 21 (31%) were part of six outbreaks. STEC O111 was isolated in 44% of all MEI. Of patients with MEI, 50% had contact with farm animals compared with 29% (P < 0.01) of persons with SEI; this difference was driven by infections involving STEC O111. More patients with non‐outbreak‐associated MEI reported drinking well water (62%) than respondents in a population survey (19%) (P < 0.01). Drinking well water and having contact with animals may be important exposures for MEI, especially those involving STEC O111.