Fig. 6: Detection of ETH2_0402100 mutation for monitoring maduramicin resistance in poultry farms.

a PCR amplification of a 306 bp fragment at the ETH2_0402100 locus using laboratory-prepared mixtures containing defined proportions (5–95%) of maduramicin-resistant E. tenella (MRR). b Amplicon sequencing of these PCR products revealed the C > T mutation at position 1255, with mutation frequencies increasing proportionally from 3 to 89%. c PCR amplification of the 306 bp fragment from 11 DNA samples collected from poultry farms. d Amplicon sequencing of field samples identified the C > T mutation at position 1255 in all eight PCR-positive farms, with mutation frequencies ranging from 1.43 to 100%.