The amount of drugs used for human or veterinary purposes is continuously growing, as well as the amount of their residues in the environment. Actually, many drugs, such as antibiotics, are poorly absorbed in the gut and are excreted as active substances. The adverse effects of these residues include toxic and genotoxic effects as well as the development of drug resistance [1], therefore requiring an attentive monitoring. Besides specific and sensitive chromatographic methods, that may require laborious sample treatment procedures [2], the presence of various xenobiotics can be easily assessed using living organisms [3] and bacteria. Among them the bioluminescent strains offer several advantages, being the light emission strictly dependent on the vitality and metabolic integrity of the bacterial cells, and the sensitivity close to that of other tests for pollutants determination [4]. A toxicological assay based on Vibrio sp. light emission was applied to the determination of antibiotic residues in excreta of pigs and turkeys treated with amoxicillin and flumequine mixture and with chloramphenicol, respectively. The residue amounts were quantified by HPLC measurements. The long term (20 h) toxicity effects were assessed by bioluminescent measurements performed using 96-well black microplates in an automatised microplate reader luminometer. The effect of standard antibiotic solutions was also determine and only Amoxicillin showed an anomalous stimulation effect till 500-800 ppm. Although a quantitative evaluation is still not possible by this assay, antibiotics extracted from real samples displayed inhibitory effects on bacterial light emission, showing a trend in good agreement with HPLC data. References: [1] Jones A.H. et al, Crit Rev Toxicol. 2004, 34 (4) 335-350. [2] S. Bogialli et al. J. Agric. Food Chem. 2004, 52, 3286-3291. [3] Wollenberger L. et al. Chemosphere, 2000, 40 (7), 723-730. [4] L.I. Sweet et al, Environ. Toxicol. Chem., 1997, 16, 2187.
Antibiotic residues revealed by bioluminescent bacteria assay / L.Bolelli; S.Bobrovová; S.Girotti; G.Fedrizzi; S.Menotta; F.Fini; E.Ferri. - STAMPA. - (2005), p. 156. (Intervento presentato al convegno RDPA- 11th Meeting on Recent Developments in Pharmaceutical Analysis tenutosi a Rimini nel 25 – 28 Settembre 2005).
Antibiotic residues revealed by bioluminescent bacteria assay
BOLELLI, LUCA;GIROTTI, STEFANO;FINI, FABIANA;FERRI, ELIDA NORA
2005
Abstract
The amount of drugs used for human or veterinary purposes is continuously growing, as well as the amount of their residues in the environment. Actually, many drugs, such as antibiotics, are poorly absorbed in the gut and are excreted as active substances. The adverse effects of these residues include toxic and genotoxic effects as well as the development of drug resistance [1], therefore requiring an attentive monitoring. Besides specific and sensitive chromatographic methods, that may require laborious sample treatment procedures [2], the presence of various xenobiotics can be easily assessed using living organisms [3] and bacteria. Among them the bioluminescent strains offer several advantages, being the light emission strictly dependent on the vitality and metabolic integrity of the bacterial cells, and the sensitivity close to that of other tests for pollutants determination [4]. A toxicological assay based on Vibrio sp. light emission was applied to the determination of antibiotic residues in excreta of pigs and turkeys treated with amoxicillin and flumequine mixture and with chloramphenicol, respectively. The residue amounts were quantified by HPLC measurements. The long term (20 h) toxicity effects were assessed by bioluminescent measurements performed using 96-well black microplates in an automatised microplate reader luminometer. The effect of standard antibiotic solutions was also determine and only Amoxicillin showed an anomalous stimulation effect till 500-800 ppm. Although a quantitative evaluation is still not possible by this assay, antibiotics extracted from real samples displayed inhibitory effects on bacterial light emission, showing a trend in good agreement with HPLC data. References: [1] Jones A.H. et al, Crit Rev Toxicol. 2004, 34 (4) 335-350. [2] S. Bogialli et al. J. Agric. Food Chem. 2004, 52, 3286-3291. [3] Wollenberger L. et al. Chemosphere, 2000, 40 (7), 723-730. [4] L.I. Sweet et al, Environ. Toxicol. Chem., 1997, 16, 2187.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
