Assessing the mobility of veterinary drugs with column experiments using different soils and under controlled flow conditions.

Veterinary pharmaceuticals are pollutants that received much attention during the last 20 years. Macrocyclic lactones are a class of drugs globally used in animal and human health, as well as crops protection. Some of its members are key substances for global food security. In this research, the mobility of eprinomectin (EPM) in soil columns (25 cm soil height; 10 cm diameter) was assessed for the first time. Soil density in the columns was 1.1-1.2 g/cm3. Porosity was 0.54-0.60. Three different soil types were used (agricultural, pastoral, wetland). In the experiment, chloride was used as a non-reactive tracer to determine the hydrodynamic conditions in the columns. Therefore, water velocity (v) was 0.146-0.151 cm/h, dispersion coefficient (D) 0.011-0.017 cm2/h and dispersivity (D/v) was 0.072-0.121. Our results showed that the drug remained in the top layers of the columns, after applying an extreme irrigation scenario. The retardation factor for EPM was 43.4-54.5 while for chloride was 0.99-1. EPM fraction (% of applied mass) in 0-1 cm was 13.8-18.0% and in 1-5 cm was 53.3-73.0%. An amount 13-29% was irreversibly bound or degraded during this experiment. From a soil management perspective, the continuous application of EPM contaminated manure, could result in high concentrations in the top 10 cm of the soil profile. Soil column experiments, where hydrodynamic conditions are well defined, are useful for the environmental impact assessment of veterinary pharmaceuticals.

Pharmacokinetics and tolerability of aminosidine after repeated administrations using an optimal dose regimen in healthy dogs and in dogs with leishmaniosis.

Optimisation of dose schedules of aminoglycosides is required in order to increase efficacy and prevent their toxicity. The objective of this study was to determine the pharmacokinetic profile and the safety of aminosidine in dogs with naturally occurring leishmaniosis and in healthy dogs after once daily administration. Six young-adult, male, healthy, Beagle dogs and 12 dogs with clinical signs of canine leishmaniosis without azotemia and proteinuria were included in the study. Diagnosis of the disease was confirmed by serology, parasitology and molecular techniques. Pharmacokinetics and evaluation of renal function after repeated (once daily for 21 consecutive days) subcutaneous administration of aminosidine, at the dose of 15 mg/kg b.w. in both the healthy and the diseased animals were compared. Concentrations of aminosidine were determined by high-performance liquid chromatography and pharmacokinetic analysis was performed by the non-compartmental method. No significant differences were observed between healthy and diseased dogs considering all pharmacokinetic parameters. In general, mean Cmax ranged between 46.41 and 54.32 µg/mL and between 38.69 and 40.73 µg/mL in healthy dogs and in dogs with canine leishmaniosis, respectively. No accumulation of the drug was observed in either group since total elimination of aminosidine and half-life lambda z were not modified throughout the administration period. Aminosidine was well tolerated in all dogs with no clinical and clinicopathological signs of nephrotoxicity. Once daily administration of high dose of aminoglycosides, resulted in effective serum concentrations and absence of nephrotoxicity.

Emergence and maintenance of multidrug-resistant Escherichia coli of canine origin harbouring a blaCMY-2-IncI1/ST65 plasmid and topoisomerase mutations.

OBJECTIVES: To characterize the mechanisms implicated in fluoroquinolone (FQ) and expanded-spectrum cephalosporin (ESC) resistance in three clinical and seven faecal multidrug-resistant (MDR; resistant to at least three antimicrobial classes) Escherichia coli isolates from a dog with atopic dermatitis, also suffering from recurrent otitis, that had already been exposed to prolonged antimicrobial treatment and colonized for a long period. METHODS: MICs of FQs, ESCs and other antimicrobials were determined by the broth microdilution method. Phenotypic tests (efflux pump inhibition and combination disc tests) and isoelectric focusing were combined with genotypic analyses [PCRs, sequencing, conjugation, S1 nuclease PFGE, PCR-based replicon typing, plasmid multilocus sequence typing (pMLST) and PCR mapping] to characterize the molecular basis of FQ and ESC resistance. Isolates were further characterized by MLST and PFGE. RESULTS: Three otitis and five faecal isolates with enrofloxacin MICs of 32 to >128 mg/L displayed the GyrA:S83L+D87N/ParC:E62K/ParE:G545D pattern harbouring novel ParC and ParE substitutions, whereas the two remaining faecal isolates were susceptible or borderline resistant single-step mutants (GyrA:S83L pattern) and carried qnrS1. Efflux pump overexpression also contributed to FQ resistance and the MDR phenotype. The three otitis and five faecal isolates also exhibited cefoxitin/ceftazidime MICs of 32-64 mg/L and harboured blaCMY-2, adjusted to ISEcp1, on an IncI1/ST65 conjugative plasmid, previously described in Salmonella Heidelberg from poultry. Interestingly, all isolates shared an identical MLST type (ST212), with the otitis isolates showing indistinguishable patterns with the high-level resistant faecal E. coli isolates. CONCLUSIONS: The long-term maintenance of FQ- and ESC-resistant clones harbouring topoisomerase mutations and a blaCMY-2-IncI1/ST65 plasmid in canine commensal flora after prolonged antimicrobial use may contribute to the dissemination of multidrug resistance.

Are the parasiticidal avermectins resistant to dissipation in the environment? The case of eprinomectin.

Eprinomectin (EPM) is a veterinary drug currently licensed in many countries for the treatment of endo- and ecto-parasites in cattle. Despite the notable evidence for its high toxicity to the terrestrial and aquatic environment ecosystems, its environmental behavior and fate are currently unknown. In the present research, the dissipation of EPM was studied in three soils and in cattle manure by using the OECD 307 guideline and the recently developed European Medicines Agency (EMA/CVMP/ERA/430327) guideline, respectively. The procedure presented by the FOrum for Co-ordination of pesticide models and their USe (FOCUS) was adopted for estimating the EPM degradation kinetics in soil and cattle manure. The EPM dissipation in soil was best described by the SFO (Simple First Order) and the HS (Hockey Stick) models, under aerobic and anaerobic conditions, respectively. The EPM dissipation in cattle manure was best described by the FOMC (First Order Multi Compartment) model. The Dissipation Time for the 50% of the initial EPM mass (DT50) range was 38-53days under aerobic and 691-1491days under anaerobic conditions. In addition, the DT50 for EPM in cattle manure was 333days. Therefore, EPM could be characterized as moderately to highly persistent to dissipation in soil, which depends on soil type, its oxygen content (aerobic or anaerobic conditions in soil) and the microbial activity. Moreover, the EPM resists dissipation in cattle manure, resulting to a high load in soil after manure application in agricultural land (or direct defecation in grassland). Consequently, the high possibility for EPM accumulation in soil and cattle manure should be considered when assessing the environmental risk of the drug.

Pharmacokinetic interaction between losartan and Rhodiola rosea in rabbits.

AIM: The study investigates the potential interaction of the herbal medicinal product of Rhodiola rosea on the pharmacokinetics of losartan and its active metabolite EXP3174 after concurrent oral administration to rabbits. MATERIALS AND METHODS: We conducted a randomized, single-dose, two-treatment, two-period, two-sequence, cross-over pharmacokinetic study on 6 healthy female New Zealand rabbits, after concurrent oral administration of losartan (5 mg/kg) and the herbal medicinal product of R. rosea (50 mg/kg). Quantification of losartan and its main active metabolite EXP3174 was achieved using a validated HPCL/UV method. Pharmacokinetic and statistical analysis was performed using the EquivTest/PK software. OBSERVATIONS: Administration of the herbal medicinal product of R. rosea resulted in a statistically significant increase of the following pharmacokinetic parameters for losartan: the maximum plasma concentration (C(max)), the area under the curve (AUC) and the apparent total body clearance (CL/F). An almost 2-fold increase in the AUC of losartan was observed after concurrent administration of the herbal medicinal product of R. rosea. No statistically significant alteration was observed in the pharmacokinetic parameters of the active metabolite of losartan EXP3174. CONCLUSION: The data of this study suggest that R. rosea significantly alters the pharmacokinetic properties of losartan after concurrent oral administration to rabbits. A study in humans should be conducted to assess the clinical significance of a possible herb-drug interaction between the herbal medicinal products of R. rosea and drugs such as losartan, which are substrates of both CYPs and P-gp.

Sorption of the antiparasitic drug eprinomectin in three soils.

Batch equilibrium studies were conducted to determine eprinomectin partitioning behavior in three Greek soils (agricultural, pastoral and riparian soil). An analytical method was developed to quantify eprinomectin in aqueous 0.01 M CaCl2. Recovery was 95% and limits of detection and quantification were both 0.005 mgL⁻¹. An existing method for its quantification in soil was successfully tested in this study. Mass balance determinations showed that we accounted for 89-98% of the eprinomectin spiked in 5 g soil/25 mL 0.01 M CaCl2. The concentration specific adsorption distribution coefficient (K(d)(ads)) ranged from 6.4 to 21.4 L kg⁻¹ while concentration specific desorption distribution coefficient (K(d)(des)) ranged from 23.2 to 124.6 L kg⁻¹. The Freundlich model adequately described adsorption and desorption with n values from 0.6 to 1.07. Hysteresis between adsorption and desorption was observed in two (agricultural and pastoral) soils. Moreover, eprinomectin binding to the clay mineral vermiculite and natural peat was tested. The drug binds to both materials. Hydroxyl groups and the nitrogen group present in eprinomectin are probably responsible for the binding to vermiculite. Coefficient K(d)(ads) significantly correlated with cation exchange capacity (CEC), Fe and Cu content of the soils when data for eprinomectin and data for ivermectin and abamectin were combined. These could be evidence that eprinomectin fate is related not only to organic matter (lipophilic binding) but also to clay content and other charged inorganic groups typically present in the soil environment.

Analytical procedure for the determination of eprinomectin in soil and cattle faeces.

A new analytical HPLC-fluorescence method was developed for the quantitative determination of eprinomectin (EPM) in soil and cattle faeces. EPM was extracted with acetone and acetonitrile from soil and cattle faeces, respectively. Solid phase extraction and derivatization reaction with N-methylimidazole in the presence of trifluoroacetic anhydride and acetic acid were applied. The limit of quantitation was 1 ng g(-1) air dried soil and 2.5 ng g(-1) moist cattle faeces. Overall recovery (RSD) was 89% (8) in soil and 85% (10) in cattle faeces and its good reproducibility (RSD<15%) allows the application of the method in advanced ecotoxicological studies, required for the environmental fate assessment of EPM.

Pharmacodynamics of amoxicillin against Mannheimia haemolytica and Pasteurella multocida and pharmacokinetic/pharmacodynamic (PK/PD) correlation in sheep.

Silicone-made tissue cages were implanted in sheep. Blood serum (SBS) and tissue cage fluid (TCF) samples were collected after amoxicillin intravenous and intramuscular administrations, at the dose of 15 mg/kg. Amoxicillin pharmacodynamics were studied in an artificial culture medium, SBS and TCF with use of a Mannheimia haemolytica and a Pasteurella multocida strain. A concentration-independent antimicrobial activity of amoxicillin was confirmed for levels higher than 0.79-1.75×MIC. This result favored the use of the percentage of the 24 h dosing interval during which drug levels remain above MIC as the appropriate pharmacokinetic/pharmacodynamic index. The subsequent correlation revealed that intravenous administration could be considered effective against "deep" infections caused by bacteria with MICs<1 µg/mL or "shallow" infections caused by bacteria with MICs<0.1 µg/mL. Intramuscular administration could be safely considered effective against both "deep" and "shallow" infections when the MICs of the targeted pathogens are lower than 1 µg/mL.

Influence of the injection site on the pharmacokinetics of amoxicillin after intramuscular administration of a conventional and a long-acting formulation in sheep.

The pharmacokinetics of amoxicillin (AMX) were investigated in sheep following intravenous (i.v.) and intramuscular (i.m) injection, comparing two different drug formulations, a conventional and a long-acting AMX-trihydrate suspension. For the i.m. application two different injections sites, the neck area and the hind limb were used to identify possible differences in the kinetic parameters related to the site of injection. A three-compartment open model could best describe AMX disposition after i.v. administration. Data analysis after i.m. administration of the conventional suspension at both injection sites revealed the occurrence of a flip-flop phenomenon, clearly indicating that absorption of AMX is the rate-limiting step of its overall disposition. A moderate effect of the injection site was observed with a tendency for the neck area to be advantageous, mainly in terms of rate rather than extent of absorption. Injection of the long-acting formulation led to a focal depot formation, thus yielding lower but remarkably prolonged serum AMX levels reflected in the respective terminal half-lives. The concentration-time profile of AMX after administration of the long-acting formulation was less affected by the injection site, but the low serum levels justify its use only in cases in which a high susceptibility of the involved bacterial population is confirmed.

Milrinone and theophylline act as lower oesophageal sphincter relaxing agents: a comparative pharmacodynamic study in the rabbit.

This study demonstrates that the inotropic agent milrinone and the bronchodilator drug theophylline exert a relaxing effect on the rabbit lower oesophageal sphincter in vitro. The relaxing effect of milrinone and theophylline, which is concentration-dependent, involves a second messenger 3',5'-cyclic adenosine monophosphate pathway and most probably it is accomplished through inhibition of phosphodiesterase (PDE) type III, as according to the obtained results it is not significantly modified either by nicotinic acid, an inhibitor of adenylate cyclase, or by the inhibitor of nitric oxide-synthetase N(omega)-nitro-L-arginine methylester and the purinergic antagonist suramin; moreover, it persists under non-adrenergic non-cholinergic conditions and it is both hexamethonium- and tetrodotoxin-insensitive. Both milrinone and theophylline display equal efficacy, comparable to that of the calcium blocker verapamil and the non-selective PDE inhibitor papaverine, but milrinone appears 50 times more potent than theophylline and three times less potent than verapamil, as, according to the pIC(50) values the potency rank of order is found to be verapamil (5.56) > milrinone (5.12) > theophylline (3.42). The here obtained pharmacodynamic profiles of the drugs suggest that both milrinone and theophylline may be considered as potent relaxing agents of the lower oesophageal sphincter.

Theophylline and its metabolites produce a stimulating cholinergic effect on the small intestine and a nonadrenergic noncholinergic relaxing effect on the colon: a comparative study in the rabbit intestine.

The present study examines comparatively the effects of theophylline and its metabolites, 1-methylxanthine (1-MX), 3-methylxanthine (3-MX), 1,3-dimethyluric acid (1,3-DMU) and 1-methyluric acid (1-MU) along the rabbit intestine, and explores the underlying mechanism(s). In the small intestine, theophylline produces atropine- and hexamethonium-sensitive increases in both the amplitude of phasic contractions and the basal tone. All metabolites mimic the theophylline's stimulating effect. In particular, concerning the phasic contractions, all metabolites are more potent than theophylline in the duodenum and jejunum, while in the ileum, only 1-MU is more potent. Regarding the basal tone, the metabolites show, in most cases, higher efficacy in all small intestinal regions, the maximum effects of 3-MX and 1-MU on the duodenum and ileum being double or triple the one of theophylline. In the ascending colon, while lower concentrations of theophylline produce an atropine- and hexamethonium-sensitive increase in the basal tone, higher ones produce a postsynaptic, nonadrenergic noncholinergic (NANC) relaxing effect. 1-MU mimics, in a weaker manner, theophylline's effect, while the other metabolites produce only relaxation, the potency rank of order being 3-MX>1-MX=1,3-DMU>theophylline. It is suggested that the theophylline and its metabolites stimulatory effect involves a cholinergic pathway, while the relaxing one is due to 3('),5(')-cyclic adenosine monophosphate (cAMP) elevation mediated by the theophylline and its metabolites inhibitory action on phosphodiesterases (PDEs).

Bioavailability and pharmacokinetics of sulphadiazine, N4-acetylsulphadiazine and trimethoprim following intravenous and intramuscular administration of a sulphadiazine/trimethoprim combination in sheep.

The combination of sulphadiazine and trimethoprim is extensively used in farm animal species; however, there are no data concerning its pharmacokinetics after intramuscular administration in sheep. Twelve rams of the Chios breed were used to study the disposition of sulphadiazine, its metabolite N4-acetylsulphadiazine and trimethoprim after intravenous (i.v.) and intramuscular (i.m.) administration of a sulphadiazine/trimethoprim (5:1) combination in sheep. Sulphadiazine bioavailability (+/-SD) was 69.00%+/-10.51%. The half-life of the terminal phase (4.10+/-0.58 h after i. v., and 4.03+/-0.31 h after i.m. administration) was significantly higher than the respective value for trimethoprim (0.59+/-0.19 h) after i.v. administration. The maintenance of a constant plasma concentration ratio after i.v. administration was therefore impossible. The acetylation capacity in sheep, determined by the AUC ratio between N4-acetylsulphadiazine and the parent compound, sulphadiazine, was very low (less than 4%). The most remarkable finding of this study was that trimethoprim was not detected in sheep plasma after i.m. injection. In conclusion, according to the findings of the present study, following i.v. administration of the sulphadiazine/trimethoprim combination, trimethoprim can be considered as the limiting factor for any possible synergistic effect, and the i.m. route cannot be recommended in sheep.

Quantitative determination of albendazole metabolites in sheep spermatozoa and seminal plasma by liquid chromatographic analysis with fluorescence detection.

A new analytical method for the simultaneous quantitative determination of albendazole metabolites in sheep spermatozoa and seminal plasma at levels down to 46.5 ng/mL for albendazole sulphoxide (ABZ-SO), 7.5 ng/mL for albendazole sulphone (ABZ-SO2) and 12 ng/mL for albendazole 2-aminosulphone (ABZ-SO2NH2) has been developed. Analytes were extracted from alkalinized samples with ethyl acetate. Separation was carried out on a C18 column in the presence of tetra-n-butylammonium (TBA) hydrogen sulphate and octanesulphonate sodium (OCT), as ion-pair agents. Fluorometric detection was performed with excitation and emission wavelengths set at 290 and 320 nm, respectively. Accuracy data showed overall recoveries (+/-S.E.M.) of 83.1+/-1.2% for ABZ-SO, 98.8+/-0.6% for ABZ-SO2 and 85.3+/-0.7% for ABZ-SO2NH2, in spermatozoa. Respective values in seminal plasma were 88.0+/-0.9%, 97.7+/-0.5% and 93.2+/-1.7%. Precision data suggested coefficient of variation (CV%) values lower than 5.9% for spermatozoa and 3.8% for seminal plasma. The method was successfully applied for the determination of the three albendazole metabolites in semen samples collected from rams that had been orally administered albendazole.

A new HPLC/UV method for the determination of clindamycin in dog blood serum.

A new HPLC method for the quantitative determination of clindamycin in dog blood serum at levels down to 80 ng/ml has been developed. Samples were deproteinised with acetonitrile and clindamycin was extracted with dichloromethane. Chromatographic analysis was carried out on a C(18) reversed-phase analytical column in the presence of tetra-n-butylammonium hydrogen sulfate (TBA), as an ion-pairing agent. UV detector wavelength was set at 195 nm. The assay was validated for a concentration range from 80 to 6000 ng/ml serum. Good linearity was observed in the entire concentration range. The limit of quantification (LOQ) was 80 ng/ml and the limit of detection (LOD) was 60 ng/ml. Regression of accuracy data yielded an overall mean recovery value (+/-S.E.M.) of 93.98+/-0.42%, while precision data revealed coefficient of variation (CV (%)) values lower than 4.41%. The method was successfully applied to determine drug concentrations in serum samples from dogs that had been orally administered clindamycin hydrochloride.

Cisapride reverses the anticholinergic effect of disopyramide on the isolated guinea-pig urinary bladder.

The present investigation aims to examine whether the prokinetic agent cisapride is able to reverse disopyramide's anticholinergic effect on the isolated guinea-pig urinary bladder. Acetylcholine, at concentrations ranging from 10(-7) to 10(-3) M, produced a stimulatory effect on the urinary bladder (pEC(50) value=5.1). Disopyramide competitively antagonized the contractile effect of acetylcholine with an ID(50)=4.4 x 10(-6) M. Although cisapride by itself had either no intrinsic contractile action or a modest effect on the urinary bladder, at concentrations ranging from 3 x 10(-7) to 10(-6) M, it significantly reversed the above inhibitory effect of disopyramide, and produced a parallel leftward shift of the concentration-response curve for acetylcholine in the presence of disopyramide. The pEC(50) values for acetylcholine in the presence of 3 x 10(-6) M and 10(-5) M disopyramide were 4.7 and 4.2, respectively, while in the presence of 10(-5) M disopyramide, after pretreatment with 5 x 10(-7) M cisapride, the pEC(50) value for acetylcholine was 4.6. It is concluded that cisapride is effective in reversing the anticholinergic activity of disopyramide on the isolated guinea-pig urinary bladder, probably by facilitating cholinergic neurotransmission.

New simple liquid chromatographic method for the determination of trimethoprim, sulfadiazine and N4-acetylsulfadiazine in plasma of broilers.

A new method for simultaneous quantification of trimethoprim, sulfadiazine and N4-acetylsulfadiazine in plasma of broilers at levels down to 13-16 ng/ml has been developed. Samples were deproteinized with acetonitrile, defatted with hexane, and extracted with dichloromethane. Chromatographic analysis was carried out on a C18 column in the presence of tetrabutylammonium hydrogen sulfate, a competing base, while detection was performed at 240 nm for trimethoprim, and 270 nm for both sulfadiazine and N4-acetylsulfadiazine. Accuracy and precision data showed recoveries and relative standard deviation values better than 87.3% and 3.1%, respectively, for all three analytes. The good analytical characteristics of the method could allow limits of detection in the low ng/ml range to be realised. The method was successfully applied to determine drug concentrations in plasma samples from broilers administered a combination of sulfadiazine and trimethoprim.

Synergistic and antagonistic pharmacodynamic interaction between ranitidine and cisapride: a study on the isolated rabbit intestine.

The present study examines the pharmacodynamic interaction between the H(2)-receptor antagonist ranitidine and the prokinetic agent cisapride on the isolated rabbit intestine. Ranitidine produced a concentration-dependent contractile effect on the duodenal, ileal and ascending colon preparations, with EC(50)values of 1.35 x 10(-4)M for the duodenum, 1.2 x 10(-4)M for the ileum and 1.15 x 10(-4)M for the ascending colon. The effect of cisapride on the ranitidine contractile effect was dependent on the cisapride concentration used. Thus, cisapride, at concentrations from 10(-10)up to 5 x 10(-7)for the duodenum and the ascending colon and up to 10(-6)M for the ileum, potentiated the contractile responses of the preparations to ranitidine. However, at higher concentrations cisapride produced a non-competitive inhibition of the intestinal contractile responses to ranitidine with IC(50)values of 4.2 x 10(-5)M for the duodenum, 1.65 x 10(-5)M for the ileum and 3.2 x 10(-6)M for the ascending colon. These data show that cisapride may modify the contractile responses of the isolated rabbit intestine to ranitidine, having a potentiating effect up to a certain concentration and an antagonistic one at higher concentrations. In conclusion, co-administration of the above drugs may lead to enhanced or reduced intestinal motility.

Cisapride counteracts the anticholinergic effect of disopyramide on the guinea pig isolated small but not large intestine.

This study examines the influence of the prokinetic agent cisapride on the disopyramide's effect on the isolated duodenum and ileum, as well as on the ascending colon of the guinea pig. Acetylcholine produced the well known stimulatory effect on the intestinal parts with EC50 values 9.6 x 10(-8), 7 x 10(-9) and 1.1 x 10(-6) M for the duodenum, ileum and ascending colon, respectively. Disopyramide inhibited, in a concentration-dependent manner, the contractile responses of the intestinal parts to acetylcholine. Cisapride, at concentrations ranging from 5 x 10(-9) M to 10(-6) M, significantly reversed the above inhibitory effect of disopyramide on the guinea pig duodenum and ileum and produced a parallel leftward shift of the concentration-response curve for acetylcholine in the presence of disopyramide. However, cisapride was not found able to reverse the inhibitory effect of disopyramide on the ascending colon. In conclusion, cisapride may counteract the inhibitory effect of disopyramide on the guinea pig duodenum and ileum, probably by facilitating the cholinergic neurotransmission, while it is not able to reverse the disopyramide's inhibitory effect on the guinea pig ascending colon.

Effect of cisapride on the isolated guinea pig gall bladder and common bile duct.

1. In this study, the effect of cisapride on the isolated guinea pig gall bladder (GB) and common bile duct (CBD) motility was investigated. 2. Cisapride, up to a certain concentration, produced an increase in tone of the GB (EC50 1.35 x 10(-8) M) and the CBD (EC50 2.75 x 10(-9) M), whereas, at concentrations higher than 3 x 10(-5) M for the GB and 5 x 10(-6) M for the CBD, it produced a transient increase in tone followed by a sustained decrease in tone or in the contraction amplitude or in both. 3. The cisapride-induced increase in tone was antagonized by atropine on both the GB and the CBD. 4. Cisapride up to 2 x 10(-5) M for the GB and 3 x 10(-6) M for the CBD did not modify, whereas, at concentrations higher than 2.8 x 10(-5) M for the GB and 4 x 10(-6) M for the CBD, it antagonized, noncompetitively, the concentration-response curve to exogenously applied acetylcholine. The IC50 values for cisapride on the EC50 of acetylcholine on the GB and the CBD were 9.4 x 10(-5) M and 8.2 x 10(-6) M, respectively. 5. In conclusion, on the guinea pig GB and CBD, low concentrations of cisapride produce a stimulating effect, probably of cholinergic origin, whereas higher concentrations produce a transient stimulating effect, probably of cholinergic origin, followed by a sustained relaxing effect, which may involve both cholinergic and noncholinergic pathways.