The isolates were characterized by PCR amplification of interd
region and mtDNA restriction analysis (RFLP-mtDNA). PCR reactions
were performed on EuroClone One Gradient thermal cycler,
using the primers d2 (50-GTGGATTTTTATTCCAACA-30) and d12 (50-
TCAACAATGGAATCCCAAC-30), as reported by Le Jeune et al. (2006).
The amplification of d regionwas performed directly fromthe colony,
without previous DNA extraction, by increasing the time and the
temperature of initial denaturation, following the protocol reported
by Capece, Romaniello, Siesto, and Romano (2012). RFLP-mtDNAwas
performed according the protocol described by Capece et al. (2010