First report of the lethal activity and synergism between deltamethrin, amitraz and piperonyl butoxide against susceptible and pyrethroid-resistant nymphs of Triatoma infestans.

In South America, Triatoma infestans (Hemiptera: Reduviidae) is the main vector of the parasite Trypanosoma cruzi, etiological agent of Chagas disease. The main strategy for vector control is to spray domestic structures with pyrethroids. Reports of populations of T. infestans with varying degrees of resistance to pyrethroids have made the search for alternative molecules for vector control necessary. In the first stage of this work we investigated the lethal activity of amitraz and deltamethrin against susceptible and pyrethroid-resistant nymphs of Triatoma infestans. Lethal dose at 50% (LD50) of susceptible nymphs were compared with those recorded in pyrethroid-resistant nymphs and the resistance ratio (RR50) was obtained. The RR50 of deltamethrin was approximately 300. In the case of amitraz, we observed similar triatomicidal activity in the two nymph populations (RR50: 0.7). In a second stage of the work, we determined the synergistic effect of amitraz and piperonyl butoxide (PBO) on the lethal activity of deltamethrin. The strong synergistic effect of PBO on the lethal activity of deltamethrin in resistant nymphs produced a decrease in RR50 to almost one third of the RR50 reported in absence of the synergist. Amitraz plus PBO lethal activity was similarly increased in pyrethroid susceptible and resistant nymphs. Our data indicate that deltamethrin synergism by amitraz was higher against resistant than to susceptible nymphs (Synergist ratio (SR50) of: 7.2- and 4.1-fold, respectively). In pyrethroid resistant nymphs, the highest level of synergism was obtained combining deltamethrin with amitraz and PBO (SR50: 26.7-fold). These results indicate that this combination could be considered an effective alternative for the control of T. infestans.

New forms of induction of apoptosis in aggressive lymphoma using peptides that interrupt the RAS / RAF interaction

Interpretation: RAS-RAF-MEK-ERK is a key pathway for apoptosis regulation in cancer cells. B-Raf-inhibitors such as PLX4032 peptide was developed by Institute Curie-Université Pierre et Marie Curie in order to induce apoptosis in cancer cells. Objectives: To demonstrate pro-apoptotic properties and survival outcome of EP2014/064243 peptide in murine aggressive lymphoma. Material and Methodology: BALBc mice with T-lymphoma were randomized assigned either in Group A (peptide+cyclophosphamide-CFM); Group B (peptides), Group C (CFM-control) or Control D (Cl-Na 0.9%-SF control group). Survival probability was calculated by Kaplan-Meier analysis. Apoptosis was detected using TUNEL technique. The protocol was approved by the Institutional Committee for Animal Care (CICUAL: T04-01-2015) Results: The median survival was 24 days (21.6-26.4) for placebo, 33 days (28.0-35.4) for the CFM monotherapy group, 33 (27.1-35.8) for the peptide group and 34 days (24,4-40) for CFM-peptide combined treatment (p<0.05). In lymph node tissue the mean TUNEL positive cells per field for each treatment group was 2, 12 and 13 and 35 for SF, CFM, peptide and combined therapy (p<0.05). Conclusion: These findings suggest that in murine aggressive lymphoma treated by an experimental peptide in addition with CFM, had an exponentially pro-apoptotic effect than CFM alone, suggesting that the peptide potentiated the anti-tumoural effect of CFM.

Pharmacokinetics of azithromycin in lactating dairy cows with subclinical mastitis caused by Staphylococcus aureus.

Azithromycin is a time-dependent antimicrobial with long persistence. The main characteristics of azithromycin suggest that it could be useful for treating bovine mastitis caused by Staphylococcus aureus. To investigate this possibility, its pharmacokinetic (PK) behavior was studied. Six Holstein lactating cows with subclinical mastitis were administered two 10 mg/kg intramuscular (i.m.) doses of azithromycin, with a 48-h interval. Milk and plasma concentrations were measured by microbiological assay. The MIC(90) was determined in 51 S. aureus isolations to calculate pharmacokinetic/pharmacodynamic (PK/PD) parameters. Milk maximal concentration (C(max)) was 7.76 +/- 1.76 microg/mL (16.67 h post-first administration) and 7.82 +/- 2.18 microg/mL (14 h post-2(nd) administration). In plasma C(max) was 0.18 +/- 0.03 microg/mL (2 h post-1(rst) administration) and 0.11 +/- 0.03 microg/mL (14 h post-2(nd) administration). Azithromycin was eliminated from the milk with a half-life (T(1/2)lambda) of 158.26 +/- 137.7 h after 2(nd) administration, meanwhile plasma T(1/2)lambda resulted shorter(13.97 +/- 11.1 h). The mean area under the concentration vs. time curve from 0 to 24 h (AUC(0-24h)) was 153.82 +/- 34.66 microg.h/mL in milk secretion and 2.61 +/- 0.59 microgxh/mL in plasma. Infection presence in the quarters had a significant effect (P < 0.05) on the area under the concentration vs. time curve from 0 to infinity (AUC(0-infinity)) and clearance from the mammary gland (Cl(mam)/F). Moreover, it had influence on milk bioavailability (F(milk)), T(1/2)lambda, AUC(0-infinity) and mean residence time (MRT) in milk, which values resulted increased in mastitic quarters. In this study, it was determined that the production level and the mammary health status have an influence on PK parameters of azithromycin treatments in bovine mastitis.

Pharmacokinetics of enrofloxacin after single intravenous administration in sheep.

The disposition of enrofloxacin in sheep was investigated after single-dose intravenous administration of 2.5 mg/kg body weight. Blood samples were drawn from the jugular vein at predetermined times after drug administration. Plasma concentrations of enrofloxacin and its active metabolite ciprofloxacin were simultaneously determined by reverse-phase high performance liquid chromatography. The data collected were subjected to non-compartmental and compartmental kinetic analysis. Statistical model theory was used to determine non-compartmental pharmacokinetic parameters. Disposition of enrofloxacin was described by a three-compartment open model with elimination from the central compartment following intravascular administration. The elimination half-life, the volume of distribution, and the area under the concentration vs time curve (AUC) were 4.31 h, 1.10 l/kg and 9.24 microg x h/ml, respectively. Enrofloxacin was metabolised to ciprofloxacin and the ratio between the AUCs of ciprofloxacin and enrofloxacin was 0.26 after intravenous administration. With predictive models of efficacy (maximum plasma concentrations/minimum inhibitory concentrations [Cmax/MIC] and AUC/MIC ratios in plasma) for most of the sheep pathogen microorganisms, enrofloxacin produced scores higher than 15 and 50, respectively. After intravenous administration atthe dose of 2.5 mg/kg, enrofloxacin achieved concentrations several times above the MIC for major pathogen bacteria in plasma, and it may prove useful in the treatment of infectious diseases caused by sensitive pathogens in sheep.

Pharmacokinetic disposition of triclabendazole in cattle and sheep; discrimination of the order and the rate of the absorption process of its active metabolite triclabendazole sulfoxide.

A comparative pharmacokinetic study was conducted to determine the order and the rate of absorption of triclabendazole (TCBZ) in cattle and sheep. A commercial suspension of TCBZ (Biofasiolex, Biogénesis S.A., Argentina) was administered at a dose rate of 10 mg/kg by the oral route to six Holstein female calves and six Corriedale female sheep. The plasma concentration profiles of the metabolites triclabendazole sulfoxide (TCBZ-SO) and triclabendazole sulfone (TCBZ-SO(2)) were analysed by means of the non-compartmental method. The order of the absorption process of the active metabolite, TCBZ-SO, was determined by construction of curves of cumulative absorbed fraction of the drug by means of the Wagner-Nelson method. The appearance of TCBZ-SO in plasma of cattle and sheep resembles the entry of a constant quantity of drug into the organism per unit time. This is explained by the reservoir effect of the rumen, which acts as a biological slow-release system for TCBZ-SO and its precursor TCBZ to the posterior digestive tract where they are absorbed. The plasma concentration profiles of TCBZ-SO in both species were well described by a one-compartment open model with zero-order process of absorption and first-order process of elimination. The values of AUC(0-infinity) and C(max) of TCBZ-SO did not differ between species, while other kinetic parameters except for lambda(z) had higher values in calves than in sheep. In the case of TCBZ-SO(2), t(max) was the only parameter that did not differ between species, while other kinetic parameters except for lambda(z) had higher values in calves than in sheep.

Correction to: T cell leukemia control via Ras-Raf pathway inhibition with peptides.

[This corrects the article on p. 172 in vol. 10, PMID: 29075346.].

Pharmacokinetics and tissue residues of an oxytetracycline/diclofenac combination in cattle.

Eight male cattle were given a combined dose containing 20 mg/kg oxytetracycline and 0.5 mg/kg diclofenac intramuscularly. Blood samples were drawn at different times until 168 h after administration. Two experimental animals were slaughtered by humane means at weekly intervals up to 28 days after administration. Samples of muscle, injection zone tissue, liver, kidney and fat were obtained. Oxytetracycline and diclofenac concentrations were determined by high performance liquid chromatography. Kinetic analysis was performed by linear regression using the CSTRIP programme. Plasma oxytetracycline concentration showed a maximum (Cmax) of 3.89 +/- 1.48 microg/ml and a prolonged elimination half-life (T1/2beta: 47.73 +/- 18.33 h). The diclofenac plasma profile showed high Cmax (577.62 +/- 238.40 ng/ml), and its T1/2beta was also prolonged (30.48 +/- 9.42 h). Oxytetracycline concentrations were measurable in liver and adipose tissue until day 21 after administration, but all tissue samples were negative for diclofenac at 21 days. The long elimination half-life of diclofenac was an unexpected finding; its T1/2beta in humans is 1.1 h.

T cell leukemia control via Ras-Raf pathway inhibition with peptides.

RATIONALE: RAS-RAF-MEK-ERK pathway has been considered a promising target for anticancer therapy. However, tumor cells may develop resistance against such drugs via hyperactivation of N-Ras, which explains why novel therapeut-ic approaches. In this sense, the Institute Curie- Université Pierre et Marie Curie (Paris 6) designed peptides in order to disturb Ras/Raf interaction which showed pro-apoptotic properties. These peptides were patented as WO2015001045 A2 (PCT/EP2014/064243)5. OBJECTIVE: In order to check the anti-tumoral action of WO2015001045 A2 peptides in a very aggressive BALB/c mice spontaneous leukemia called LB, we performed the present study. METHOD & RESULTS: 50 BALB/c mice inoculated with 106 LB tumor cells were randomly assigned either to control (placebo) or treatment group (that daily received 3 mg of peptide per kg of mice) during 30 days. By day 15 only 24% of the control group was alive vs. 100% of the treatment group. The average survival in treated group was 20,27 days while in control group the mean survival was 15,48 days. Either bone marrow, spleen or axillary nodes demonstrated a higher level of malignant T cell presence compare with treated group (89,78% ; 95,64% & 77,68% versus 72,45%, 80,23% & 63.44% respectively for each organ inspected. DISCUSSION: Our study demonstrated an improvement in survival curves in mice model affected by spontaneous T lymphoid leukemia when peptides WO2015001045 A2 were used. These peptides might be a valid option to become part of the therapeutic armory for malignant lymphoproliferative diseases control.

Pharmacokinetics of ricobendazole after its intravenous, intraruminal and subcutaneous administration in sheep.

Ricobendazole (RBZ) was administered in sheep at the dose rate of 5 mg/kg by intravenous (i.v.) route as a 10% experimental solution, by the intraruminal (i.r.) route as a 10% experimental suspension, and by the subcutaneous (s.c.) route as a 10% commercial formulation available in Argentina. Blood samples were drawn during a 60 h period. Plasma concentrations of RBZ and its inactive metabolite albendazole sulphone (ABZSO2) were determined by high-performance liquid chromatography. The pharmacokinetic parameters were determined by compartmental analysis. The fitting of the data was done by weighted least-squares non-linear regression analysis. The pharmacokinetic parameters were estimated for every animal by simultaneous fitting of the plasma concentrations profiles of RBZ obtained after its administration by the three routes. The kinetic analysis of ABZSO2 was performed by a statistical moment approach. Ricobendazole bioavailability was poor after i.r. administration, whereas high and sustained plasma concentrations and higher bioavailability were obtained after s.c. administration. A simple two-compartment open model explains in a mechanical sense the pharmacokinetic behaviour of RBZ in sheep and allows us to estimate the real first-order constant rate of absorption and the loss of drug from the absorption site after its administration by s.c. and i.r. routes.

Efficacy of injectable doramectin in the therapy and control of Dermatobia hominis infestations in Latin America.

Three studies were conducted in Latin America, one in Brazil, one in Venezuela and one in Argentina, using a common protocol to investigate the efficacy of a single subcutaneous injection of doramectin at 200 micrograms kg-1 (1 ml per 50 kg) for the treatment and control of Dermatobia hominis infestations in cattle raised under commercial conditions. In each study, two groups of 20 animals each were allocated on the basis of D. hominis nodules present 24 h before treatment to a treated group (T1), or to a control group (T2) which received saline solution at 1 ml per 50 kg of live weight. All cattle were injected in the mid-dorsal cervical region and examined on treatment day and 2, 7, 15 and 30 days post-treatment (p.t.). At each observation day the number of D. hominis nodules was counted and the viability of the larvae inside them was assessed and recorded. Treatment with doramectin was 100% effective in eliminating D. hominis larvae and in controlling the re-establishment of the parasite under field conditions of continuous reinfestation. During the first 48 h p.t., dead larvae of different ages were found outside or partially outside the nodules in animals of the doramectin groups. After 48 h p.t., no live larvae were found inside existing nodules, nor did new nodules develop in animals of the doramectin groups in any of the three studies. When compared with nodule counts in the control group on the same observation day or with parasite burdens on the same animal before treatment, doramectin-treated animals began to eliminate parasitic nodules at 48 h p.t. Efficacy reached 100% at 7 days p.t. and remained at 100% on subsequent observation days (P < 0.05). None of the doramectin-treated animals exhibited any clinical signs of adverse reaction to medication.

Efficacy of doramectin in the protection of neonatal calves and post-parturient cows against field strikes of Cochliomyia hominivorax.

Two studies were conducted in Latin America, one in Venezuela and one in Argentina, using a common protocol to investigate the efficacy of a single s.c. injection of doramectin in the prevention and control of field infestations of Cochliomyia hominivorax in newborn calves and post-parturient cows. In both experiments, pregnant cows were paired on the basis of their calving time. The first cow that calved was assigned to a medicated group, and both cow and calf were treated with doramectin. The cow received a dose of 200 micrograms kg-1 and the calf 1 ml of a 1% doramectin solution. The second cow and calf were assigned to a saline-treated group, and the cow received saline at 1 ml per 50 kg and the calf 1 ml. The procedure was repeated until all animals were allocated to the two treatments. Animals were examined on the treatment day and at 2, 4, 6 and 12 days post-treatment. At each observation day, the navel of newborn calves and external genitalia of the cows were inspected and the presence of C. hominivorax infestations was recorded. Doramectin was 100% effective in the prevention and control of screwworm strikes in newborn calves and in post-parturient cows exposed to continuous field challenge of C. hominivorax. Over the 12 day duration of the studies, 48% and 57% of the saline-treated calves had screwworm navel strikes in the studies in Venezuela and Argentina, respectively. Infested animals required repeated therapeutic treatment, whereas none of the doramectin-treated calves were affected (P < 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)

Comparative persistent efficacy of doramectin, ivermectin and fenbendazole against natural nematode infections in cattle.

A study was conducted in Argentina, to investigate the period of protection of a single injection of doramectin administered subcutaneously (s.c.) at 200 micrograms kg-1 (1 ml/50 kg) compared with single treatments of ivermectin (200 micrograms kg-1 s.c.) and fenbendazole (5 mg kg-1 p.o.), against field infections of gastrointestinal parasites of cattle. Eighty-three animals were selected and ranked on the basis of serial fecal egg counts (e.p.g.'s). From this group, three animals were slaughtered before treatment and their lungs, abomasum, small and large intestines, were processed for parasite counts and identification. The remaining 80 animals were allocated in ranked groups of four to a control or one of three treated groups. Animals of the four groups were grazed together in the same pasture for the duration of the study. Treatments were administered on Day 0. Individual fecal samples were collected at weekly intervals for the first 49 days post-treatment and twice a week from Day 52 to Day 84 (end of study). At each collection day fecal samples were pooled for coprocultures. On Day 28 and 56, two animals from each group, previously identified on Day 0, were killed and their parasite burdens determined. The duration of protection of a single injection of doramectin was longer than ivermectin or fenbendazole treatment. On Day 56, the total number of parasites found in doramectin-treated animals was significantly (P < 0.05) lower than parasite burdens found in either ivermectin- or fenbendazole-treated animals. The longer persistent activity of doramectin was expressed by the lower number of adults and L4 stages of Ostertagia ostertagi. Data from this experiment demonstrated the limitations of using fecal egg counts to evaluate the persistent efficacy of anthelmintics. The duration of activity of doramectin was demonstrated more accurately by parasite counts in cattle from each group since decreasing e.p.g.'s were seen in non-medicated animals without changes in total parasite burdens.

Efficacy of injectable doramectin in the protection of castrated cattle against field infestations of Cochliomyia hominivorax.

Three studies were conducted in Latin America--one in Venezuela, one in Argentina and one in Brazil--using a common protocol to investigate the efficacy of a single subcutaneous injection of doramectin in the prevention and control of Cochliomyia hominivorax infestations in castrated cattle. In each study, two groups of 20-28 animals each were allocated to a treated (T1) or to a control (T2) group on the basis of body weights. Animals of T1 received doramectin at 200 micrograms kg-1 (1 ml per 50 kg) and animals of T2 received saline solution at 1 ml per 50 kg of live weight. After treatment all cattle were castrated surgically. Animals were examined on treatment day and at 2, 4, 6 and 12 days post-treatment. At each observation day, the presence of C. hominivorax infestations was recorded. Doramectin was 100% effective in the prevention and control of screwworm strikes in castrated cattle exposed to continuous field infestations of C. hominivorax in tropical and subtropical areas of Latin America. Over the 12 day duration of the studies, 85%, 60% and 65% of animals in the control groups had infested wounds in Venezuela, Argentina and Brazil, respectively. Affected animals required repeated therapeutic treatment, whereas none of the doramectin-treated cattle were infested (P < 0.0001). A high proportion of the castration wounds in doramectin-treated cattle had the presence of characteristic C. hominivorax eggs but none developed into larvae. There were no clinical signs of adverse reactions to treatment in any of the three studies.

Tissue concentrations of a long-acting oxytetracycline formulation after intramuscular administration in cattle.

The tissue distribution of a long-acting oxytetracycline formulation after intramuscular administration to calves at a dose rate of 20 mg/kg was studied. Oxytetracycline concentrations were determined in lung, bone marrow, mammary gland, uterus, uterine horn, ovary, liver, synovial fluid, joint tissue, kidney, spleen, brain, muscle, fat, urine, bile, saliva, ruminal content and serum. Observed concentrations were higher than the minimal inhibitory concentration for the majority of pathogens in all of the analysed tissues for at least 72 h post-injection. The shortest theoretical permanency time was 1.88 days (in saliva) and the longest was 19.06 days (in kidney). Taking the maximal permitted residue level as 0.1 microgram/g in muscle, 0.3 microgram/g in liver and 0.6 microgram/g in kidney, the calculated withdrawal time was 20 days.

Efficacy of injectable doramectin against natural Boophilus microplus infestations in cattle.

Three experiments were conducted in Latin America--1 in Brazil, 1 in Venezuela, and 1 in Argentina--using a common protocol to investigate the efficacy of a single SC injection of doramectin (200 micrograms/kg of body weight) for treatment and control of Boophilus microplus infestations in grazing cattle. In each experiment, 2 groups of 20 to 27 cattle were allocated on the basis of tick burdens present 24 hours before treatment to a treated or control group, which received saline solution (1 ml/50 kg). All cattle were injected in the mid-dorsal cervical region and examined on treatment day and on posttreatment days (PTD) 8, 14, 21, and 28. On each observation day, the number of engorged and semiengorged ticks on half of each animals was counted and recorded. Doramectin was highly efficacious in removing tick burdens and controlling reestablishment of B microplus under field conditions of continuous reinfestation. Compared with tick counts in the control group, doramectin-treated cattle began to eliminate ticks after treatment and efficacy improved on subsequent observation days. In Brazil, efficacy was 91% at PTD 8 and 14, and increased to 99% or higher at PTD 21 and 28. In Venezuela, efficacy was 100% at PTD 8, 14, and 21 and 92% at PTD 28. In Argentina, efficacy progressed from 94.5% at PTD 8 to 99 and 98.5% at PTD 21 and 28, respectively. The reduction of the number of ticks in the doramectin-treated cattle was significant (P < 0.05) at each observation day.(ABSTRACT TRUNCATED AT 250 WORDS)

A discriminatory study of a pharmacokinetic model for intramuscular gentamicin in sheep.

The resulting serum concentrations were measured in six ewes after intramuscular administration of 10 mg/kg of gentamicin. The model providing the best fit for the experimental data was determined both by linear regression analysis between the experimental and theoretical values and by means of the Minimum Akaike Information Criterion Estimation (MAICE) test. Linear regression analysis showed certain differences favouring the monocompartmental model although the advantage was not conclusive. The MAICE test, however, permitted a clear discrimination in favour of the same model. When linear regression analysis is not conclusive, the MAICE test represents a good alternative.

The pharmacokinetics of a slow-release theophylline preparation in horses after intravenous and oral administration.

The pharmacokinetics of a slow-release theophylline formulation was investigated following intravenous and oral administration at 10 mg/kg in horses. A tricompartmental model was selected to describe the intravenous plasma profile. The elimination half-life (t1/2 beta) was 16.91 +/- 0.93 h, the apparent volume of distribution (Vd) was 1.35 +/- 0.18 L/kg and the body clearance (ClB) was 0.061 +/- 0.009 L kg-1 h. After oral administration the half-life of absorption was 1.24 +/- 0.30 h, and the calculated bioavailability was above 100%. The t1/2 beta after oral administration was 18.51 +/- 1.75 h, only a little longer than that after intravenous administration. The slow release formulation did not exhibit any advantage in prolonging the t1/2 beta of theophylline in the horse.

The effects of the method of calculation on the evaluation of the pharmacokinetic parameters of oxytetracycline after intravenous administration to calves.

The aim of this study was to assess the differences in the values of the pharmacokinetic parameters attributable to the use of either linear or nonlinear regression analysis and to find the effect of weighting schemes on these differences. Six calves received 20 mg/kg oxytetracycline i.v. Blood samples were drawn during 72 h. The assay of the drug was performed microbiologically. A bicompartmental pharmacokinetic model was used, kinetic analysis being carried out by linear regression (LR) and by weighted least-squares nonlinear regression (WLSNLR). Statistical analysis included a test for normality, the Kruskall-Wallis test and ANOVA with log transformation. The A0, alpha and B0 did not show any statistically significant differences attributable to the mathematical method used. On the other hand, the statistically significant differences in the beta values found using the Kruskall-Wallis test and ANOVA with log transformation could be attributed to the different methods employed. ANOVA with log transformation determined statistically significant differences between the parameters obtained by linear analysis and those obtained by WLSNLR when the weighting (w) was 1. When weights were 1/x, 1/x2 or 1/square root of x, no statistically significant differences were found. The optimal weighting scheme was w = 1/x2 because of a more homogeneous scatter and random distribution of residuals about the abcissa axis in a plot of weighted residuals in the ordinate versus time in the abcissa. It was concluded that the use of these different procedures can give major variations in the apparent value of beta, the most important pharmacokinetic parameter. The correct selection of the weighting procedure is therefore fundamental in obtaining the best estimate of this pharmacokinetic parameter in WLSNLR.

The pharmacokinetics of thiamphenicol in lactating cows.

The pharmacokinetics of thiamphenicol were studied after intravenous and intramuscular administration of 25 mg/kg body weight in lactating cows. Distribution (t1/2 alpha) and elimination (t1/2 beta) half-lives of 6.10 +/- 1.39 min and 1.60 +/- 0.30 h, respectively, were obtained after intravenous administration. The body clearance was 3.9 +/- 0.077 ml/kg per min and the apparent volume of distribution was 1220.79 +/- 256.67 ml/kg. The rate at which thiamphenicol appeared in the milk, as indicated by the penetration half-life (t1/2P) (serum to quarters), was found to be 36.89 +/- 11.14 min. The equivalent elimination half-life (t1/2E) (quarters to serum) from the milk was 3.62 +/- 1.06 h and the peak thiamphenicol concentration in the milk was 23.09 +/- 3.42 micrograms/ml at 2.5 +/- 0.32 h. After intramuscular injection, the elimination half-life was 2.2 +/- 0.40 h, the absorption half-life was 4.02 +/- 1.72 min and the peak concentration in the serum was 30.90 +/- 5.24 micrograms/ml at 23 +/- 8.4 min. The bioavailability after intramuscular administration approached 100%. The penetration half-life was 50.59 +/- 6.87 min, the elimination half-life was 5.91 +/- 4.97 h and the mean peak concentration in the milk was 17.37 +/- 2.20 micrograms/ml at 3.4 +/- 0.22 h.

Effect of 1-(1-naphthylmethyl)-piperazine on antimicrobial agent susceptibility in multidrug-resistant isogenic and veterinary Escherichia coli field strains.

The objective of this study was to evaluate the interaction of the efflux pump inhibitor 1-(1-naphthylmethyl)-piperazine (NMP) when combined with different families of antimicrobial agents against isogenic strains and multidrug-resistant (MDR) Escherichia coli field strains isolated from animals. Laboratory isogenic strains of E. coli with different levels of expression of efflux pumps were used as quality controls. Ten MDR E. coli strains were collected from healthy animals in a cross-sectional study in four commercial dairy farms. The MICs of florfenicol, ciprofloxacin, tetracycline and ampicillin were determined by a serial microdilution method in Luria-Bertani broth in the presence or absence of NMP. NMP used with ampicillin exerted no effect on the isogenic or field strains. In most of the field MDRE. coli strains and in an acrAB-overexpressing (AG112) isogenic strain, the MICs of florfenicol, ciprofloxacin and tetracycline decreased at least fourfold when the antimicrobial was combined with the highest NMP concentrations. In the wild-type strain (AG100), there were no decreases of more than twice the MIC, whilst in strain AG100A, an efflux pump-deficient strain, the MIC did not change, regardless of the concentration of NMP used with these three antimicrobials. Thus, ampicillin was not affected by the efflux pump mechanism, whereas ciprofloxacin, tetracycline and florfenicol were shown to be substrates of efflux pumps, with a consequent significant reduction in MICs. Resistance could not be completely reversed in the E. coli field strains by NMP, probably because other resistance mechanisms were also present. However, in strain AG112, the MIC results demonstrated that NMP expressed an important synergistic activity with florfenicol. The reduction in florfenicol MIC value was sufficient to reverse antimicrobial resistance completely for AG112.