Genotoxicity of AMPA, the environmental metabolite of glyphosate, assessed by the Comet assay and cytogenetic tests.

Formulations containing glyphosate are the most widely used herbicides in the world. AMPA is the major environmental breakdown product of glyphosate. The purpose of this study is to evaluate the in vitro genotoxicity of AMPA using the Comet assay in Hep-2 cells after 4h of incubation and the chromosome aberration (CA) test in human lymphocytes after 48h of exposition. Potential in vivo genotoxicity was evaluated through the micronucleus test in mice. In the Comet assay, the level of DNA damage in exposed cells at 2.5-7.5mM showed a significant increase compared with the control group. In human lymphocytes we found statistically significant clastogenic effect AMPA at 1.8mM compared with the control group. In vivo, the micronucleus test rendered significant statistical increases at 200-400mg/kg. AMPA was genotoxic in the three performed tests. Very scarce data are available about AMPA potential genotoxicity.

HPLC residues of enrofloxacin and ciprofloxacin in eggs of laying hens.

Eggs of 12 laying hens with 5 mg/kg/day oral administration of 5% enrofloxacin (EFX) or ciprofloxacin (CFX) solution during 5 days contained residues from 0.02 to 1.98 microg/g (EFX) or 0.14 to 0.28 microg/g (CFX). At identical dosage regime High Performance Liquid Chromatograhy (HPLC) residues of EFX were 6-fold greater than CFX ones. Maximun concentrations were detected at the second day after the administration withdrawal. The limits of detection were 0.019 microg/g for EFX and 0.156 microg/g for CFX. The recovery was 36-50% for CFX and 49-85% for EFX. The withdrawal treatment periods in hens are six days for EFX and five days for CFX in order to avoid violative levels of egg residues.

Intravenous pharmacokinetics of ciprofloxacin in goats.

The pharmacokinetics of ciprofloxacin in goats was studied following intravenous administration. A single dose of 10 mg/kg was administered as an intravenous bolus, and serum samples were collected at predetermined intervals over a 24 h period. Ciprofloxacin levels were measured using high pressure liquid chromatography and the resulting concentration versus time curve was analyzed using a non linear regression analysis. The data were best represented by a two-compartment pharmacokinetic model with a mean elimination half-life of 2.72+/-1.04 h. Mean pharmacokinetic parameters obtained were area under the curve (AUC: 10.320+/-5.137 microg/ml per h), mean residence time (MRT: 3.334+/-1.428 h), apparent volume of distribution (Vdss: 3.373+/-0.893 l/kg) and total body drug clearance (tCl 19.596+/-9.059 ml/min per kg). Ciprofloxacin in goats showed the general pharmacokinetic characteristics of a typical fluoroquinolone antibacterial agent and we recommend a dose of 10 mg/kg to be administered intravenously at 12 h intervals to goats.

Chromosomal aberrations in human lymphocytes exposed in vitro to enrofloxacin and ciprofloxacin.

Chromosomal aberrations were evaluated in cultures of human peripheral lymphocytes from eight healthy donors, exposed to the antimicrobial enrofloxacin (EFX) or to its major metabolite ciprofloxacin (CFX). In both treatments cultures revealed an increase in the chromosomal aberration level, detected as chromatid and chromosome breaks and gaps. Control cultures analysis revealed 3.6 +/- 0.6 chromosomal aberrations per 100 cells while treated cultures exhibited 8.3 +/- 0.8 and 9.6 +/- 1.2 aberrations at 5 and 50 microg/ml of EFX respectively. In CFX treated cultures it was found 5.6 +/- 1.3 and 7.7 +/- 3.5 aberrations/100 cells at 5 and 25 microg/ml antimicrobial concentration. These results suggested a genotoxic effect of EFX and CFX in the system used (P < 0.001). A reduction in the mitotic index and fuzzy metaphases were observed at 50 microg/ml of CFX, indicating a cytotoxic effect produced by this antimicrobial.

Increase of tissue lipid hydroperoxides as determination of oxidative stress.

Increased levels of lipid hydroperoxides (LOOH) are frequently associated with the oxidative mechanisms involved in physiological states as ageing and with serious pathological conditions. In the present work the physiological and the CCl4-induced lipid hydroperoxides levels in mice liver and kidney were determined. The analysis of LOOH tissue levels was performed through the oxidation of 1-napthyldiphenylphosphine (NDPP) into its oxide (ONDPP) and further quantification by high pressure liquid chromatography at 292 nm UV detection. The physiological level of lipid hydroperoxides levels was higher in the kidney (245 +/- 8 nmol LOOH/g of tissue) than in liver (164 +/- 5 nmol of LOOH/g tissue). After a single administration of CCl4 (0.25 ml/g) tissue LOOH reached a maximum level after 15 min (416 +/- 21 nmol/g kidney and 303 +/- 6 nmol/g liver) and then slowly decreased. LOOH levels in liver afforded an early indicator (15 min) of oxidative damage. LOOH levels in kidney remained significatively increased up to 60 min post administration. The described HPLC assay is a useful, simple and sensitive method to detect cellular oxidative stress and damage.

Comparative pharmacokinetics of enrofloxacin and ciprofloxacin in chickens.

The pharmacokinetics of enrofloxacin (EFX) and ciprofloxacin (CFX) was investigated in broiler chickens. Each antimicrobial was administered intravenously at a dose of 5 mg/kg body weight. Blood was taken in different preset times: prior and at 0.03, 0.06, 0.13, 0.25, 0.5, 1, 2, 4, 8, 12 and 24 h following drug administration. The concentrations of EFX and CFX in plasma were determined by high pressure liquid chromatography (HPLC). Plasma concentrations vs. time were analysed by a compartmental independent pharmacokinetic model that provided the most important kinetic parameters. Statistically significant differences between the two antimicrobials were found for most of the pharmacokinetic parameters: Area under the curve (AUC), area under first moment curve (AUMC), mean residence time (MRT), total body cleareance (ClB), volume of distribution beta (Vd beta) and volume of distribution at the steady state (Vd(ss)). Both antimicrobials were widely distributed in chickens throughout the body with a mean Vd(ss) of 1.98+/-0.18 L/kg for EFX, and 4.04+/-0.69 L/kg for CFX. The ClB for CFX was five times higher than that obtained for EFX. AUC, MRT and the diminished half time for EFX were two-four times higher than those obtained for CFX. These results indicate that CFX remains in the body for less time than the other quinolone. This characteristic of CFX suggests the advantage of a shorter withdrawal time for food producing animals treated with this antimicrobial.