Collaborative study of a microbiological screening method (three-plate) for the banned antimicrobial growth promotors tylosin, virginiamycin, spiramycin, zinc bacitracin and avoparcin in animal feed

Abstract

A microbiological screening method (three-plate) for the detection of the antimicrobial growth promoters tylosin, spiramycin, virginiamycin, zinc bacitracin, and avoparcin in animal feed has been developed and validated successfully. A collaborative study involving 18 laboratories receiving 172 samples was carried out to verify the performance characteristics. The detection level for tylosin/virginiamycin/spiramycin, expressed in microbiological activity, was 1 mg kg(-1) (false-positives, 2%; false-negatives, 3, 0, and 6%, respectively). Avoparcin could be detected at 1 mg kg(-1) in feed in general (false-positives, 2%; false-negatives, 0%). However, in calf feed the sensitivity was lower. The percentages of false-negatives were found to be 12%, 7%, and 0% at 1, 3, and 5 mg kg(-1), respectively (false-positives, 4%). The limit of detection for zinc bacitracin was 3-5 mg kg(-1) (false-positives, 5-10%; false-negatives, 77% at 1 mg kg(-1), 45% at 2 mg kg(-1), 12% at 3 mg kg(-1), and 4% at 5 mg kg(-1)). The method allowed for a distinction to be made between the groups of antibiotics: avoparcin/zinc bacitracin versus tylosin/virginiamycin/spiramycin. This definitely gives added value to the method in the framework of a follow-up of positive screening results by post-screening and confirmatory analysis.

Simultaneous detection of tylosin and tilmicosin in honey using a novel immunoassay and immunochromatographic strip based on an innovative hapten

“…Several methods have been developed for monitoring TYL and TIM residues, such as fluorescence immunoassay (Bang-Ce, Songyang, Peng, & Xiao-Hong, 2008;Su et al,. 2011;Wei et al, 2013), microbiological assay (Litterio, Calvinho, Flores, Tarabla, & Boggio, 2007;Nouws et al, 1999;Pol-Hofstad, Driessen-Van Lankveld, Tomassen, De Jong, & Van Egmond, 2008), electrochemical enzyme-linked immunosorbent assay (ELISA) (Ammida et al, 2004;Draisci et al, 2001), thin-layer chromatography coupled to microbiological detection (Vincent Gizzi, Holst, Jong, & Michard, 2007), high-performance liquid chromatography (HPLC) (Clark, Dowling, & Boison, 2009;Prats, Francesch, Arboix, & Perez, 2002;Zheng et al, 2011), and HPLC coupled with mass spectrometry (HPLC-MS) (Juan, Moltó, Mañes, & Font, 2010;Nozal Nalda et al, 2006;Thompson, Noot, Calvert, & Pernal, 2003). However, most of these assays are time consuming and require expensive equipment, specialized technical personnel, and complicated sample pre-treatment and, thus, are unsuitable for extensive screening and field detection.…”

Time-resolved fluoroimmunoassay for quantitative determination of tylosin and tilmicosin in edible animal tissues

“…For animal product safety control, several detection methods for tylosin and tilmicosin residues have been developed in recent years, including high performance liquid chromatography (HPLC) [4][5][6][7][8], thin-layer chromatography [9], HPLC coupled with mass spectrometry [10][11][12][13][14][15], and microbiological assays [4,16]. However, expensive equipment, skilled technicians and complicated sample pretreatments are required for those methods, making them unsuitable for high throughput field detection or routine screening.…”