A Critical Review of Screening Methods for the Detection of Chloramphenicol, Thiamphenicol, and Florfenicol Residues in Foodstuffs

This article gives an extensive overview of the wide range of analytical procedures developed for the detection of amphenicol antibiotic residues (chloramphenicol, thiamphenicol, and florfenicol) in many different types of foodstuffs (milk, meat, eggs, honey, seafood). Screening methods such as microbial inhibition methods, antibody-based immunoassays using conventional and biosensor-based detection systems, and some methods based on alternative recognition systems are described. The relative advantages and disadvantages of these methods are discussed and compared. The current status and future trends and developments in the need for accurate and rapid detection of this group of antimicrobials are also discussed.

Eficácia do florfenicol e da oxitetraciclina no controle de Aeromonas hydrophila em pacu (Piaractus mesopotamicus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

ACUTE TOXICITY and ENVIRONMENTAL RISK of OXYTETRACYLINE and FLORFENICOL ANTIBIOTICS TO PACU (Piaractus mesopotamicus)

The oxytetracyline (OTC) and florfenicol (FLO) are the most used antibiotics for bacterial disease control in fish. The aims are: estimating the lethal concentration 50% initial (LC(I) 50;48h) of OTC (Terramicina (R)) and FLO (Aquaflor (R)) for pacu (Piaractus mesopotamicus); classifying OTC and FLO according to the acute toxicity and the risk of environmental intoxication; and assessing OTC and FLO dilution effect in water quality variables. Fish were acclimatized for 10 days in a bioassay room at 27 +/- 2 degrees C. Then nine animals (three each concentration) were exposed to 7.0; 7.5; 8.0 or 8.5 mg L(-1) of OTC or to 600.0; 700.0; 800.0; 900.0 and 1000.0 mg L(-1) of FLO and a control treatment. The water quality variables recorded daily were: pH, temperature, electric conductivity, and dissolved oxygen. OTC and FLO were classified according to acute toxicity and environmental risk. The estimated OTC LC(I) 50;48h for pacu is 7.6 mg L(-1) and the estimated FLO is > 1000 mg L(-1). OTC reduced the dissolved oxygen concentration, and it was considered moderately toxic, causing risk of environmental intoxication to pacu. FLO is not toxic; it does not cause risk of environmental intoxication to pacu and does not change water quality variables.

Determinação simultânea de resíduos de cloranfenicol, tianfenicol e florfenicol em leite bovino por cromatografia eletrocinética micelar

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efficacy of florfenicol and intravenous fluid therapy for treatment of experimental salmonellosis in newborn calves

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ecotoxicidade e eficácia da oxitetraciclina e do florfenicol contra infecção experimental por Aeromonas hydrophila e aspectos histopatológicos em pacu (Piaractus mesopotamicus)

Pós-graduação em Aquicultura - FCAV

Avaliação do perfil farmacocinético do florfenicol em plasma bovino após aplicação intramuscular de duas doses e avaliação da sua eficácia a bactérias sensíveis

Pós-graduação em Ciência Animal - FMVA

Genotoxic responses of juvenile tilapia (Oreochromis niloticus) exposed to florfenicol and oxytetracycline

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Eficácia do florfenicol e da oxitetraciclina no controle de Aeromonas hydrophila em pacu (Piaractus mesopotamicus)

Determinou-se a concentração eficaz de oxitetraciclina (OTC) e florfenicol (FFC) no tratamento de Aeromonas hydrophila em pacu (Piaractus. mesopotamicus). Os pacus foram submetidos à captura duas vezes ao dia por quatro dias e em seguida foram infectados com A. hydrophila (2,4x10(7) bactéria mL-1). Os tratamentos utilizados foram: controle sem infecção (CSI), controle com infecção (CCI) e tratados com 110,0; 140,0 e 170,0mgOTC.kg-1, e 5,0; 10,0 e 15,0mgFFC.kg-1. As variáveis de qualidade da água foram monitoradas diariamente. Após o tratamento, no CSI dos dois testes, ocorreu 100% de sobrevivência. Nos testes com OTC, no CCI, a sobrevivência foi de 29,2%; em 110,0mg.kg-1, 37,5%; em 140,0mg.kg-1, 29,2%; e em 170,0mg.kg-1, 50,0%. Nos testes com FFC, foi eficaz com 10,0mg.kg-1, e no CCI a sobrevivência foi de 76,9%; em 5,0mg.kg-1, 81,81%; em 10,0mg/L.kg-1, 100% e em 15,0mg.kg-1, 87,5%. A OTC, em concentrações de até 170,0mg.kg-1 de ração, não é eficaz para o controle de A. hydrophila em pacu, e o FFC é eficaz na concentração de 10,0mg.kg-1 e ambos não alteram as variáveis de qualidade de água.

Antibiotic Sensitivity Testing

Respiratory bacterial pathogens in pigs are currently treated with antibiotics. Intervet - Schering Plough markets an antibiotic called Nurflor (Florfenicol) targeting respiratory pathogens. This project tests the effectiveness of this antibiotic against a series of respiratory pathogens. 6 isolates will be tested per serovar/strain and the isolates will be from 4 different farms using MIC testing. The sensitivity of Florfenicol will be compared to sensitivity of the organisms to Tilmicosin and Amoxicillin. Development of resistance to certain antibiotics have been reported, so it is important to have alternative antibiotics available to treat the respiratory pathogens on farms.

Antimicrobial susceptibility of Histophilus somni isolated from clinically affected cattle in Australia

This study investigated antimicrobial resistance traits, clonal relationships and epidemiology of Histophilus somni isolated from clinically affected cattle in Queensland and New South Wales, Australia. Isolates (n = 53) were subjected to antimicrobial susceptibility testing against six antimicrobial agents (ceftiofur, enrofloxacin, florfenicol, tetracycline, tilmicosin and tulathromycin) using disc diffusion and minimum inhibitory concentration (MIC) assays. Clonal relationships were assessed using repetitive sequence PCR and descriptive epidemiological analysis was performed. The H. somni isolates appeared to be geographically clonal, with 27/53 (47%) isolates grouping in one cluster from one Australian state. On the basis of disc diffusion, 34/53 (64%) isolates were susceptible to all antimicrobial agents tested; there was intermediate susceptibility to tulathromycin in 12 isolates, tilmicosin in seven isolates and resistance to tilmicosin in one isolate. Using MIC, all but one isolate was susceptible to all antimicrobial agents tested; the non-susceptible isolate was resistant to tetracycline, but this MIC result could not be compared to disc diffusion, since there are no interpretative guidelines for disc diffusion for H. somni against tetracycline. In this study, there was little evidence of antimicrobial resistance in H. somni isolates from Australian cattle. Disc diffusion susceptibility testing results were comparable to MIC results for most antimicrobial agents tested; however, results for isolates with intermediate susceptibility or resistance to tilmicosin and tulathromycin on disc diffusion should be interpreted with caution in the absence of MIC results.

Characterisation of antibodies to chloramphenicol, produced in different species by enzyme-linked immunosorbent assay and biosensor technologies

Six polyclonal antisera to chloramphenicol (CAP) were successfully raised in camels, donkeys and goats. As a comparison of sensitivity, IC50 values ranged from 0.3 ng mL(-1) to 5.5 ng mL(-1) by enzyme-linked immunosorbent assay (ELISA) and from 0.7 ng mL(-1) to 1.7 ng mL(-1) by biosensor assay. The introduction of bovine milk extract improved the sensitivity of four of the antisera by ELISA and two by biosensor assay; a reduction in sensitivity of the remaining antisera ranged by a factor of 1.1-2.6. Porcine kidney extract reduced the sensitivity of all the antisera by a factor ranging from 1.1 to 7 by ELISA and a factor of 1.5 to 4 by biosensor. A low cross-reactivity with thiamphenicol (TAP) and florfenicol (FF) was displayed by antiserurn G2 (1.2% and 18%, respectively) when a homologous ELISA assay format was employed. No cross-reactivity was displayed by any of the antisera when a homologous biosensor assay format was employed. Switching to a heterologous ELISA format prompted three of the antisera to display more significant cross-reactivity with TAP and FF (53% and 82%, respectively, using Dl). The heterologous biosensor assay also increased the cross-reactivity of D1 for TAP and FF (56% and 129%, respectively) and of one other antiserum (Gl) to a lesser degree. However, unlike the ELISA, the heterologous biosensor assay produced a substantial reduction in sensitivity (by a factor of 6 for D1). (C) 2007 Elsevier B.V. All rights reserved.

A competitive enzyme-linked immunosorbent assay for determination of chloramphenicol

Chloramphenicol is a broad-spectrum antibiotic shown to have specific activity against a wide variety of organisms that are causative agents of several disease conditions in domestic animals. Chloramphenicol has been banned for use in food-producing animals for its serious adverse toxic effects in humans. Due to the harmful effects of chloramphenicol residues livestock products should be free of any traces of these residues. Several analytical methods are available for chloramphenicol analysis but sensitive methods are required in order to ensure that no traces of chloramphenicol residues are present in edible animal products. In order to prevent the illegal use of chloramphenicol, regulatory control of its residues in food of animal origin is essential. A competitive enzyme-linked immunosorbent assay for chloramphenicol has been locally developed and optimized for the detection of chloramphenicol in sheep serum. In the assay, chloramphenicol in the test samples and that in chloramphenicol-horseradish peroxidase conjugate compete for antibodies raised against the drug in camels and immobilized on a microtitre plate. Tetramethylbenzidine-hydrogen peroxide (TMB/H2O2) is used as chromogen-substrate system. The assay has a detection limit of 0.1 ng/mL of serum with a high specificity for chloramphenicol. Cross-reactivity with florfenicol, thiamphenicol, penicillin, tetracyclines and sulfamethazine was not observed. The assay was able to detect chloramphenicol concentrations in normal sheep serum for at least 1 week after intramuscular injection with the drug at a dose of 25 mg/kg body weight (b.w.). The assay can be used as a screening tool for chloramphenicol use in animals.

Characterization of antimicrobial resistance in Aeromonas and Vibrio isolated in Canada from fish and seafood

Plusieurs études ont examiné la sensibilité aux antimicrobiens chez les bactéries d’organismes provenant de produits issus de l’aquaculture ou de leur environnement. Aucune information n’est cependant disponible concernant la résistance aux antimicrobiens dans les bactéries de la flore de poissons ou de fruits de mer vendus au détail au Canada. C’est particulièrement vrai en ce qui a trait aux bactéries des genres Aeromonas et Vibrio, dont certaines espèces sont des agents pathogènes zoonotiques connus. Au cours de cette étude, la sensibilité aux antimicrobiens d’isolats d’Aeromonas spp. et de Vibrio spp. provenant de poissons et de crevettes domestiques et importés a été mesurée à l’aide de techniques de micro dilution en bouillon et/ou de diffusion sur disque. Les classes d’antimicrobiens examinés comprenaient les tétracyclines (TET), les inhibiteurs de la voie des folates (sulfadiméthoxine-triméthoprime, SXT), le florfenicol (FLO), et les quinolones (acide nalidixique / enrofloxacine, NA/ENO). Des valeurs seuils épidémiologiques pour Aeromonas et Vibrio ont été établies en utilisant la méthode d’interprétation normalisée des données de résistance provenant de diffusion sur disque. La recherche de gènes de résistance associés au profil de résistance des isolats a été effectuée en utilisant des PCRs et des puces ADN. Le nombre d’isolats résistants aux divers antimicrobiens parmi les 201 isolats d’Aeromonas et les 185 isolats de Vibrio étaient respectivement les suivants: TET (n=24 et 10), FLO (n=1 et 0), SXT (n=2 et 8), NA (n=7 et 5) et ENO (n= 5 et 0). Diverses associations de gènes tet(A), tet(B), tet(E), floR, sul1, sul2, et intI1 ont été détectées, les gènes tet(E), intI1, sul2 et tet(B) étant les plus communs. Les espèces d’Aeromonas et de Vibrio isolées de poissons au détail et de fruits de mer peuvent héberger une variété de gènes de résistance, bien que peu fréquemment. Le risque que représente ces gènes de résistance reste à évaluer en considérant le potentiel infectieux des bactéries, l’utilisation des ces agents antimicrobiens pour le traitement des maladies en aquaculture et en médecine humaine et leur rôle en tant que réservoir de la résistance antimicrobienne.