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REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228

Volume 14 - Número 2 - 2º Semestre 2014

RESIDUES OF ENROFLOXACIN AND CIPROFLOXACIN IN POUL TRY TISSUES

Arnildo Korb1; Eleusis R. de Nazareno2; Paulo R. Dalsenter3; Ciro A. de O. Ribeiro4; Constança Pomba5

ABSTRACT We investigated the existence of residues of the antimicrobials enrofloxacin and ciprofloxacin in broilers, which are exposed to the consumer in the metropolitan city Curitiba, Brazil. Antimicrobial residues in animal tissues, above the minimum allowed concentration, pose risks to human health. In 10 samples analyzed by Liquid Chromatography (Mass Spectometry), at the Lanagro laboratory, from Ministry of Agriculture, Livestock and Supply of Brazil, considering a lower limit of 20 µg.kg-1 and maximum 100 µg.kg-1, no residues of ciprofloxacin and enrofloxacin were detected. These results showed that in the broiler samples of different brands, these antimicrobials were not detected. Keywords: Antimicrobial residues, poultry tissues, ciprofloxacin and enrofloxacin. RESÍDUOS DE ENROFLOXACINA E CIPROFLOXACINA EM TECID OS DE FRANGOS RESUMO Investigou-se a existência de resíduos dos antimicrobianos enrofloxacina e ciprofloxacina em frangos expostos ao consumo em um município na região metropolitana de Curitiba, Brasil. Resíduos de antimicrobianos em tecidos animais acima dos limites permitidos (LMR) representam riscos à saúde humana. Em 10 amostras analisadas pelo laboratório Lanagro, do Ministério da Agricultura Pecuária e Abastecimento do Brasil, analisadas por Cromatografia Líquida Acoplada à Espectrometria de Massas, pela metodologia utilizada, não foram detectados resíduos dentro do limite mínimo de 20 µg.kg-1 e máximo de 100 µg.kg-1 permitido. Palavras chave: Resíduos de antimicrobianos, tecidos de frangos, ciprofloxacina e enrofloxacina.

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INTRODUCTION Enrofloxacin is one of the most common antibiotics used in animal production (LOLO et al, 2006; REYES-HERRERA et al, 2011) to control Escherichia coli and Salmonella (HARITOVA et al, 2011) infection. The main metabolite of this antimicrobial is ciprofloxacin (GARCIA et al, 2001; GARCIA et al, 2005), exclusively prescribed for treating severe infections in humans (BALL, 2000), especially those caused by β-lactam antibiotic-resistant bacteria (RUIZ et al, 2012). Antibiotic residues in products of animal origin are observed when veterinary practices are not followed, for example, when administering higher dosages than those prescribed, using products for longer than the recommended duration, and using a non-indicated route of administration (MICHELL et al, 1998). When present in food for human consumption, antibiotic residues in concentrations beyond the allowable limits may present some toxicological effects such as allergies, in addition to contributing to the selection of resistant bacteria in human gut microbiota, thereby compromising antibiotic therapies (LINDER et al, 2012; KIM et al, 2012). The maximum residue levels in foods allowed in Brazil are in accordance with Codex Alimentarius (Food and Agriculture Organization) and are authorized by the European Community. The monitoring control policies on food residues are established by the Normative Instruction DAS/MAA 42/1999 of the Ministry of Agriculture, Livestock and Supply and ratified by Normative Instruction no. 7 of March 27, 2013. The Ministry of Agriculture, Livestock and Supply, through the laboratory Lanagro, is responsible for conducting liquid chromatographic analyses of chicken tissue samples collected by federal inspection bodies to verify the presence of quinolone residues. Publications reporting research results on antibiotic residues in chickens distributed for human consumption are either lacking or limited. METHODOLOGY

In December 2011, a survey of major chicken brands sold for human consumption was conducted in Fazenda Rio Grande, in the metropolitan region of Curitiba. Seventeen brands were identified in supermarkets and meat markets. To sample each of the 10 predominant brands, namely Jaguá, Canção, Maringá, Goldfrango, Diplomata, Bigfrango, Aurora, Anhembi, Sadia, and Cooperave, one frozen chicken was purchased and 100 grams of its breast was removed. The samples were frozen at −20ºC and sent to the laboratory of the Ministry of Agriculture, Livestock and Supply (Lanagro), in the city of Porto Alegre, for liquid chromatographic analysis coupled with mass spectrometry. The analysis followed the internal protocol (method MET RPM/07/02), with minimum and maximum residue quantification levels of 20 and 100 µg/kg, respectively, as adopted by the National Plan for Control of Residues and Contaminants. The methodology used by Lanagro follows Directive 2002/657/EC of the European Union, which establishes performance indications, criteria, and procedures for analytical methods.

RESULTS AND DISCUSSION

The method used by Lanagro did not

detect, in the 10 samples tested, any residues with concentrations within the allowable levels.

A study on chickens distributed for human consumption in supermarkets and meat markets in the state capital of Brazil, conducted in 2011 by the Office of Consumer Protection in São Paulo, also did not detect any antibiotic residues, including enrofloxacin and ciprofloxacin, with concentrations within the allowable limits (IDEC, 2011). CONCLUSION

Publications regarding the presence of

antibiotic residues in animal tissues for human consumption in Brazil are lacking.

At least for the chicken tissue samples and antibiotics assessed in this study, we can confirm that regulations regarding toxicological aspects in terms of allowable minimum and

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maximum residue levels in meat for domestic consumption are followed by agribusinesses in Brazil. BIBLIOGRAPHIC REFERENCES BAILAC S.; BARRÓN D.; SANZ-NEBOT V.; BARBOSA J. Determination of fluoroquinolones in chicken tissues by LC-coupled electrospray ionisation and atmospheric pressure chemical ionisation. J. Sep. Sci. v.29, n.1, p. 131 – 136, 2006. BALL, P. Quinolone generations: natural history or natural selection? Journal of Antimicrobial Chemotherapy. v.46, n.1, p.17-24, 2000. GARCIA, M. A; SOLANS, C.; HERNANDEZ, E.; PUIG, M.; BREGARTE, M. A. Simultaneous determination of enrofloxacin and its primary metabolite, ciprofloxacin in chicken tissues. Cromatographia. v.54, n.3/4, p.191-194, 2001. GARCIA, M.A.; SOLANS, C.; CALVO, A.; HERNANDEZ, E.; REY, R.; BREGANTE, M.A.; PUIG, M. Determination of Enrofloxacin and its primary metabolite, Ciprofloxacin, in pig tissues. Application to residue studies. Biomedical Chromatography, v.19, n.1, p.27-31, 2005. HARITOVA, A.; URUMOVA, V.; LUTCKANOV, M; PETROV, V.; LASHEV, L. Pharmacokinetic-pharmacodynamic indices of enrofloxacin in Escherichia coli O78/H12 infected chickens. Food and Chemical Toxicology. v.49, p.1530–1536, 2011. INSTITUTO BRASILEIRO DE DEFESA DO CONSUMIDOR-IDEC. Produto Seguro, produção obscura. Revista 151. Fev.2011 Disponível l em: http://www.idec.org.br/em-acao/revista/151/materia/produto-seguro-produco-obscura Acesso em: 15/02/2012, 2011. KIM, B-S.; KIM, J. N.; YOON, S-H; CHUN, J; CERNIGLIA, C.E. Impact of enrofloxacin on the human intestinal microbiota revealed by comparative molecular analysis. Anaerobe. V.18, n.18, 310-320, 2012.

LINDER, S W.; VEACH, B.T.; YAN, S.S, A; FERNÁNDEZ, H.; PINEIRO, S. A., AHN Y.; CERNIGLIA, C. E. In vitro enrofloxacin binding in human fecal slurries. Regulatory Toxicology and Pharmacology. v.62, n.1, 74–84. 2012. LOLO, M.; PEDREIRA, S.; MIRANDA, J. M.; VA´ZQUEZ, B. I.; FRANCO, C. M.; CEPEDA, A.; FENTE, C. Effect of cooking on enrofloxacin residues in chicken tissue. Food Additives and Contaminants. v.23, n.10, 988–993. 2006. MITCHELL, J.M.; GRIFFITHS, M.W.; MCEWENS, S.A; MCNAB, W.B; IEE L. Antimicrobial drug residues in milk and meat causeas, concerns prevalence, regulations, testx and test performance. Journal of food protection. v.61, n.6, p.742-756, 1998. REYES-HERRERA, I.; SCHNEIDER, M.J.; BLORE, P.J.; DONOGHUE, D. J. The relationship between blood and muscle samples to monitor for residues of the antibiotic enrofloxacin in chickens. Poultry Science. v.90, n.2, p. 481–485, 2011. RUIZ, J.; PONS, M.J; GOMES, C. Transferable mechanisms of quinolone resistance. International Journal of Antimicrobial Agents. v.40, n. 3, p. 196-203, 2012. _____________________________________ 1-Nursing Department of the Santa Catarina State University (UDESC), Rua Licério Sperry, 233 D, Bairro Engenho Braun, CEP 89809360, Chapecó, SC, Brazil. PhD student in Environment and Development. CAPES fellow. arkorb@yahoo.com.br 2-Department of Collective Health of the Federal University of Paraná (UFPR) 3-Pharmacology Department of the Federal University of Paraná (UFPR) 4-Toxicology Department of the Federal University of Paraná (UFPR) 5-Faculty of Veterinary Medicine, University of Lisbon (ULisboa)