Pig Urine Research Articles

Species differences in metabolism of codeine: urinary excretion of codeine glucuronide, morphine-3-glucuronide and morphine-6-glucuronide in mice, rats, guinea pigs and rabbits

1. Metabolites of codeine were determined by use of h.p.l.c. in urine of male mice, rats, guinea pigs and rabbits injected with 10 mg codeine/kg subcutaneously. 2. In 24 h urines of these species, unchanged codeine, codeine glucuronide, free morphine, and morphine-3-glucuronide were as follows: mice, 6.8, 1.6, 0.8 and 7.6% dose; rats, 1.6, 0.2, 4.3 and 23.9% dose; guinea pigs, 1.6, 39.8, 0.2 and 1.6% dose; rabbits, 2.2, 24.5, 1.3 and 17.9% dose. Urinary excretion of morphine-6-glucuronide was 0.7% dose in guinea pigs, 1.9% in rabbits, and not detectable in mice and rats. Norcodeine was found only in the urine of mice. 3. These results indicate that codeine is metabolized in all four species by glucuronidation and by oxidative N- and O-demethylation, but the quantitative excretions of metabolites were quite different in different species.

実験動物アレルギーの臨床的，免疫学的研究 とくにモルモット上気道アレルギーを中心に

With the increasing numbers of individuals who take care animals, there has been a significant increase in the incidence of allergy to laboratory animals, and this has become an important social problem.The present study was carried out to clarify the mechanism of LAA, to identify the major allergens in guinea pig urine and saliva, and to find a way to prevent allergic symptoms.The patients were apt to have positive skin tests, high levels of serum specific IgE and immediate positive reactions to intranasal allergen challenge tests. LAA therefore seems to be a type 1 allergy. The molecular weights of major allergens determined by polyacrylamide gel electrophoresis and gel chromatography were 14, 000 to 18, 000 daltons in guinea pig urine and 15, 000 to 52, 000 daltons in guinea pig saliva. The results of RAST inhibition assays showed cross-allergenicity among guinea pig urine and saliva and serum and pelt and rat urine.A new type of air conditioning with better air flow (laminary air flow in one direction) than in the conventional turbulent flow system appeared to be effective for one of the ways to prevent LAA.

Direct estimation of estrone sulfate in serum and urine of pregnant swine as indicators of litter size at birth

Previous studies have suggested that estrone sulfate (E 1S) concentrations in maternal serum or urine of pigs are correlated with litter size. Therefore the goals of this study were to determine the following: (1) are E 1S concentrations in maternal blood and urine on day 24 or 28 of pregnancy positively correlated with litter size at birth? (2) are E 1S concentrations in maternal blood and urine correlated? and (3) how useful are E 1S measurements in maternal blood or urine as a predictor of litter size at birth? Estrone sulfate was measured by direct radioimmunoassay in serum and urine of pregnant sows on days 24 and 28 postcoitum. The levels of E 1S in serum and urine were higher on day 28 than on day 24. On day 24, neither maternal serum nor urinary E 1S concentrations were correlated with total litter size at birth. On day 28, both serum ( r = 0.47; P < 0.001) and urinary ( r = 0.33; P < 0.04) E 1S levels were correlated with litter size at birth. Moreover, levels of urinary and serum E 1S were correlated ( r = 0.56; P < 0.001) on day 28 but not on day 24. Although relationships exist between litter size at birth and the levels of E 1S in serum and urine on day 28, quantitative predictions of litter size using serum or urinary E 1S levels are relatively imprecise.

Natural occurrence and preparation of O-acylated 2,3-unsaturated sialic acids

Three O-acylated, unsaturated sialic acids, N-acetyl-9- O-acetyl, N-acetyl-9- O-lactoyl-, and 2-deoxy- N-glycoloyl-9- O-lactoyl-2,3-didehydroneuraminic acid (5-acetamido-9- O-acetyl-, 5-acetamido-9- O-lactoyl-, and 2,6-anhydro-3,5-dideoxy-5-glycoloylamido-9- O-lactoyl- d- glycero- d- galacto-non-2-enonic acid) were isolated from urine or submandibular glands of rat, pig, and cow. Mass spectrometric evidence for the existence of 2,3-unsaturated 9- O-acetyl- N-glycoloylneuraminic acid in porcine urine was also obtained. The sialic acids were purified by dialysis, gel- and ion-exchange chromatography, and preparative thin-layer chromatography. They were analyzed by thin-layer chromatography, high-pressure liquid chromatography, and capillary gas-liquid chromatography-mass spectrometry. For comparison, O-acetylated unsaturated sialic acids were synthesized.

Detection of leptospires in biological fluids using DNA hybridisation

DNA extracted from Leptospira interrogans serovar pomona was labelled with phosphorus-32 by nick translation and used as a genomic probe to detect leptospiral DNA. The sensitivity of detection in a 10-μl spot on nylon membranes was 160 pg of leptospiral DNA or 1.1 × 10 3 leptospires and assays with nylon membranes were somewhat more sensitive than assays with nitrocellulose membranes. The probe reacted with the pathogenic hardjo and tarassovi leptospiral serovars, but not with other genera of bacteria. To detect leptospires in body fluids, these were treated to free leptospiral DNA and then concentrated on membranes using a Bio-Dot™ apparatus. Neither serum nor urine interfered with the assay system. The DNA of leptospires added to pig urine was stable for at least 2 h at room temperature and for at least 20 h at −20°C.

Variations in the monoamine oxidase-inhibitory activity ("tribulin?") in pig's urine.

Sandler and his colleagues (see Sandler, 1982) have demonstrated the presence of an endogenous inhibitor of the enzyme monoamine oxidase (MAO) and of benzodiazepine receptor binding in the urine and blood plasma of man and rat. The concentrations of this material increased under stress conditions and it has been named "tribulin". In the present experiments MAO-inhibitory activity was found in extracts of urine and plasma samples of domestic pigs. Evidence was obtained that the inhibitory activity was higher when pigs experienced slight discomfort. Thus it appears that pigs produce a substance similar to tribulin. It may become possible to use such MAO-inhibitory activity as an indicator in the assessment of interaction with the environment in pig husbandry.

Analytical methods for the study of urinary thioether metabolites in the rat and guinea pig

Methods are described for the isolation and identification of three classes of bivalent sulfur metabolites characterized as neutral methylthio ethers, ethyl acetate-soluble acidic thioethers and ethyl acetate-insoluble acidic thioethers from rat and guinea pig urine. After extraction of the metabolites by the ammonium carbonate—ethyl acetate procedure, the individual metabolites are separated by capillary gas chromatography and/or by high-performance liquid chromatography with both μBondapak C 18 and Porasil columns. Identification of the metabolites is based on gas chromatography—mass spectrometry (electron impact) and on fast atom bombardment mass spectrometry. Interesting species differences in metabolism were observed. The major ethyl acetate-soluble acidic thioethers in rat urine are mercapturic acids. In contrast, in the guinea pig a new pathway involving mercaptopyruvic, mercaptolactic and mercaptoacetic acids is operative. The thioether metabolites of styrene oxide and phenanthrene are described, but the procedures have been applied in studies of several drugs and environmental chemicals in our laboratory.

Absorption, distribution, metabolism and excretion of a new morpholine derivative Y-8894 in mice, rats, guinea pigs and rabbits.

Absorption, distribution, metabolism and excretion of a new morpholine derivative Y-8894 were investigated in mice, rats, guinea pigs and rabbits by administration of 14C labeled compound (14C-Y-8894) at a dose of 30 mg/kg. Urinary and fecal recoveries of radioactivity (14C) within 4 days following oral administration were 52.2 % and 46.8 % in mice, 24.3 % and 72.5 % in rats, 73.4 % and 22.5 % in guinea pigs, and 64.8 % and 26.0 % in rabbits, respectively. About 93 % of the given 14C was recovered in the rat bile within 24 hours after oral administration. Maximum blood levels of 14C were obtained within 1 hour after oral administration in all species examined, with all initial biological half-lives (T 1/2) in the range of 1.0 ?? 1.8 hours. Terminal T1/2s were 21.2 ?? 25.8 hours for all species except the guinea pigs. The blood levels in guinea pigs were not detectable 24 hours after oral administration. The highest levels of 14C were observed in the liver, kidney and lung of mice, and in the liver of rats 0.5 hours after oral administration, and relatively high levels in the lung and kidney of rats, in the adrenal, pancreas, salivary gland and spleen of both mice and rats. Brain levels in both mice and rats were twice of that in the blood. Twenty-four hours after administration significant amount of 14C were detected only in the liver and kidney. Protein bindings of 14C in the serum were in the range of 63 ?? 83 % for all species. Major metabolites of Y-8894 in urine of mice were similar to those in urine and bile of rats, but somewhat different from those in urine of guinea pigs and rabbits.

Method for determination of morphinone in urine and bile of guinea pig by high-performance liquid chromatography

Method for determination of morphinone in urine and bile of guinea pig by high-performance liquid chromatography

Analytical methods for the study of urinary thioether metabolites in the rat and guinea pig

Methods are described for the isolation and identification of three classes of bivalent sulfur metabolites characterized as neutral methylthio ethers, ethyl acetate-soluble acidic thioethers and ethyl acetate-insoluble acidic thioethers from rat and guinea pig urine. After extraction of the metabolites by the ammonium carbonate—ethyl acetate procedure, the individual metabolites are separated by capillary gas chromatography and/or by high-performance liquid chromatography with both μBondapak C 18 and Porasil columns. Identification of the metabolites is based on gas chromatography—mass spectrometry (electron impact) and on fast atom bombardment mass spectrometry. Interesting species differences in metabolism were observed. The major ethyl acetate-soluble acidic thioethers in rat urine are mercapturic acids. In contrast, in the guinea pig a new pathway involving mercaptopyruvic, mercaptolactic and mercaptoacetic acids is operative. The thioether metabolites of styrene oxide and phenanthrene are described, but the procedures have been applied in studies of several drugs and environmental chemicals in our laboratory.

Catecholamines in the Vitreous Fluid and Urine of Guinea Pigs Dying of Cold and the Effect of Postmortem Freezing and Autolysis

Concentrations of catecholamines in vitreous fluid and urine in guinea pigs dying of cold and the effects of freezing and autolysis on these parameters were studied. The analysis was performed by high performance liquid chromatography with electrochemical detection. Noradrenaline (NA) concentration in vitreous fluid was more than 20 times higher in the cold exposed animals than in controls (44.2 +/- 9.2 versus 2.0 +/- 1.0 ng/mL). Autolysis alone caused an increase to 33.5 +/- 7.7 ng/mL, and freezing alone to 13.4 +/- 5.3 ng/mL. The highest values were in the group with exposure, freezing, and autolysis. Adrenaline (A) concentration in the vitreous fluid increased fourfold (3.9 +/- 1.5 versus 0.7 +/- 0.5 ng/mL) in cold exposure and twofold as a result of autolysis. Dopamine (DA) concentration in vitreous fluid was elevated only in the group with exposure, freezing, and autolysis. The increase of NA concentration in urine was fivefold during the whole exposure (from 19.4 +/- 6.9 to 109 +/- 57.3 ng/mL), but A was increased by twentyfold (from 10 +/- 5.1 to 213.2 +/- 168.7 ng/mL), whereas DA concentration did not change. The increase of average excretion of NA to urine was eightfold during the first 6 h of exposure, and that of A tenfold. According to the present results, elevated concentrations of catecholamines in the vitreous fluid and urine can be used as a diagnostic aid for hypothermia death. Concerning the values of noradrenaline in the vitreous, the increase as a result of autolysis must be taken in account when interpreting the results.

Gas Chromatographic Determination of Avilamycin Total Residues in Pig Tissues, Fat, Blood, Feces, and Urine

A gas chromatographic (GC) procedure is presented for the determination of residues of avilamycin and all its metabolites/conjugates which can be converted to the common moiety dichloroisoeverninic acid (DIA). The method involves alkaline hydrolysis to DIA, cleanup by partitioning with chloroform, acidification of the aqueous phase, and partitioning of DIA into methylene chloride. After methylation of DIA, the product, 3,5-dichloro-4,6-dimethoxy-2-methylbenzoic acid methyl ester, is cleaned up on a silica gel column prior to the final determination by electron capture GC. The method is sensitive to 0.1 mg/kg avilamycin equivalent. Overall average recoveries were 85.4%, with a standard deviation of 9.1% for n = 20. Analyses of feces, urine, tissues, and fat of pigs treated with avilamycin demonstrated that 93% of the administered substance is excreted in feces and urine, within 72 h after treatment, and that no residues (less than 0.01 mg/kg) can be found in the tissues and fat of the animals at any time between 0 and 7 days after treatment with medicated feed.

Structural parameters and natural occurrence of 2-deoxy-2,3-didehydro-N-glycoloylneuraminic acid.

2-Deoxy-2,3-didehydro-N-glycoloylneuraminic acid has been found to occur in porcine, bovine and equine submandibular glands as well as in the urine of pig, horse and rat. This novel, unsaturated sialic acid was isolated by gel filtration and ion-exchange chromatography. Final purification was achieved by column chromatography or by preparative thin-layer chromatography on cellulose. The structural analysis was performed by combined capillary gas-liquid chromatography/mass spectrometry. The various data were compared with those from synthetic 2-deoxy-2,3-didehydro-N-glycoloylneuraminic acid. Besides of the unsaturated N-glycoloylated sialic acid, also the corresponding N-acetylated derivative was present in the materials analyzed. The inhibitory effect of 2-deoxy-2,3-didehydro-N-glycoloylneuraminic acid on Vibrio cholerae sialidase using N-acetylneuraminyl-(alpha 2----3)-lactose as substrate is slightly higher (50% inhibition at 10 microM) when compared with 2-deoxy-2,3-didehydro-N-acetylneuraminic acid (50% inhibition at 15 microM).

Alterations in concentrations of cyclic guanosine 3′,5′-monophosphate in guinea pig urine during the development of a transplantable leukaemia

Cyclic guanosine 3′,5′-monophosphate (cGMP) concentrations were measured in urine specimens from guinea pigs before and after transplantation of a leukemia. A transient increase in cGMP concentration occurred 3 days after inoculation which preceded any detectable increase in the white blood cell count. This peak in cGMP concentration was found to be very highly significant when compared with fluctuations in urinary cGMP levels in the guinea pigs from transplantation of the leukemia. Inoculation of guinea pigs with irradiated L2C cells suggests that transplantation may cause an initial depression in urine cGMP concentration but no elevated cGMP concentrations were observed in these animals. The results indicate that urine cGMP concentrations may be a useful way of monitoring abnormal cell proliferation.

Enzymatic radioimmunoassay for detecting Leptospirainterrogans serovar Pomona in the urine of experimentally-infected pigs

An enzymatic radioimmunoassay (ERIA) has been developed for detecting Leptospira interrogans serovar pomona in porcine urine. Four grower pigs were experimentally infected with serovar pomona. A total of 39 urine samples was collected, and ERIA was compared with dark ground microscopy (DGM) and culture for demonstrating leptospiruria. Of 20 samples positive by at least one technique, leptospires were detected by ERIA in 14, by culture in 16 and by DGM in 13. ERIA, unlike the other 2 methods, was suitable for use with urine which had been stored frozen for several months.

Metabolism of Meperidine in Several Animal Species

Four new meperidine metabolites were identified by GC-MS in the urine of rats, guinea pigs, rabbits, cats, and dogs. In addition to known meperidine metabolites, 4-ethoxycarbonyl-4-phenyl-1,2,3,4-tetrapyridine dehydronormeperidine; IV, the N-hydroxydehydro derivative of normeperidine (X), the dihydroxy derivative of meperidine (XII), and the dihydroxy derivative of normeperidine (XIII) were identified. The possible role of the N-hydroxy derivative of normeperidine (IX) in the pharmacological interaction of meperidine (I) with MAO inhibitors, seen selectively in the rabbit (and humans), is discussed. Following the administration of the p-hydroxy derivative of meperidine (VII), the major metabolite was conjugated VII. Trace amounts of the p-hydroxy derivative of normeperidine (VIII), the methoxy hydroxy derivative of meperidine (XI), XII, and XIII also were detected as metabolites of VII. The degree of N-demethylation of VII, both in vitro and in vivo, was small.

Comparative metabolism of the cis and trans isomers of N-nitroso-2-6-dimethylmorpholine in rats, hamsters and guinea pigs

The in vivo metabolism of the cis and trans isomers of N-[3,5- 3H] nitroso-2,6-dimethylmorpholine (NDMM) was studied in female Fischer rats, Syrian golden hamsters and guinea pigs by analysis of urinary metabolites using high pressure liquid chromatography (HPLC). Animals were treated by gavage with 12 mg/kg body wt. of NDMM, composed of both isomers and 12 μCi/kg body wt. of either of the separated radioactive isomers ( cis or trans). Control animals received 12 mg, 12 μCi/kg body wt. NDMM with both isomers labeled in their natural proportion. There was a substantial increase in the excretion of a particular metabolite, 2-(2-hydroxyl-methyl)ethoxy propanoic acid, in the urine of rats, hamsters and guinea pigs 24 h after received the trans isomer (24, 22 and 13% of the total dose excreted, respectively). A minor metabolite was determined to be 2,6-dimethylmorpholine-3-one, another product of α-oxidation. The metabolite 1-amino-2-hydroxypropanol was identified, indicating that NDMM was metabolized by both α-and β-oxidation. In all three species, animals administered the cis isomer excreted larger amounts of N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine (HPOP) and N-nitroso-bis(2-hydroxypropyl)amine (BHP) products of beta oxidation, than those treated with the trans isomer. Hamsters and guinea pigs treated with the more carcinogenic cis isomer in these species, also excreted twice as much of two other metabolites than was found in the urine of animals given the trans isomer. The trans isomer of NDMM appeared to be preferentially metabolized by α-oxidation and from earlier studies this metabolic pathway seemed to be important in carcinogenesis by NDMM in the rat. The cis isomer might be in a conformation more favorable for β-oxidation and this pathway may be of primary importance in carcinogenesis by NDMM in hamsters and guinea pigs.

Guinea-pig-derived allergens. Clinicoimmunologic studies, characterization, airborne quantitation, and size distribution.

Of 20 serum samples from laboratory workers who experienced allergic symptoms after exposure to laboratory animals, 9 contained elevated IgE antibodies to guinea pig urine, pelt, or albumin by the radioallergosorbent test (RAST). Skin testing documented allergenic activity with all 3 guinea pig allergens. The cross-reactivity of these 3 guinea pig allergens was studied in RAST inhibition assays. Guinea pig pelt extract and urine produced dose-related inhibition in all 3 assays, but guinea pig albumin was inhibitory in only the homologous assay. Characterization by gel filtration, polyacrylamide gel electrophoresis, and isoelectric focusing demonstrated heterogeneity of the allergens. Air filter samples were taken in a guinea pig housing room, a research laboratory, and a library, using a high volume air sampler. An outdoor air filter sample was also taken. Guinea pig urine allergen activity was detected in all indoor air filter samples by RAST inhibition. Guinea pig pelt allergen activity was detected in all air filter samples, except the library and outdoors. Guinea pig albumin was not detected in any air filter sample. Sizing of airborne particles demonstrated that most guinea pig allergen activity was associated with particles of a diameter greater than 5 mu and less than 0.8 mu. Urine appears to be the major source of guinea pig allergens, and it is present in airborne particles small enough to penetrate the lower respiratory tract when inhaled.

Determination of trimethoprim metabolites including conjugates in urine using high-performance liquid chromatography with combined ultraviolet and electrochemical detection

A high-performance liquid chromatographic method for the determination of trimethoprim metabolites in pig urine was developed. The metabolites — glucuronic acid and sulphuric acid conjugates of phenolic metabolites formed by demethylation of trimethoprim — were quantitated after treatment of urine with β-glucuronidase ( Escherichia coli). The sulphuric acid conjugate was not susceptible to enzymatic hydrolysis and was therefore assayed as the conjugate by use of ion-pair chromatography on the reversed-phase column. In order to find suitable conditions for enzymatic hydrolysis of the glucoronides, the conjugates were obtained by gel chromatography of urine from a [ 14C]trimethoprim-treated pig.

Metabolism of p-tert.-butyltoluene in the rat and guinea pig.

The metabolism of p-tert.-butyltoluene (TBT) was studied in the rat and guinea pig. Both the methyl and the tert.-butyl group were oxidized to alcohol and carboxylic acid derivatives in these species. The major urinary metabolites in rats were p-tert.-butylbenzoic acid and its alcohol derivative 2-(p-carboxyphenyl)-2-methylpropan-1-ol whereas p-tert.-butylbenzoylglycine was the most prominent metabolite in guinea pig urine. No significant differences in metabolism were found when TBT was given intragastrically or by inhalation. The intragastric administration of 14C-TBT to rats showed that the bulk of the excretion of radioactivity occurred within three days. A recovery of 83% was achieved and the ratio of urinary/faecal radioactivity was roughly 3.5:1.