Tsukamurella hongkongensis sp. nov. and Tsukamurella sinensis sp. nov., isolated from patients with keratitis, catheter-related bacteraemia and conjunctivitis

Tsukamurella serpentis sp. nov., isolated from the oral cavity of Chinese cobras (Naja atra)

“…We also recently reported the discovery of two novel species of the genus Tsukamurella, Tsukamurella hongkongensis and Tsukamurella sinensis, from patients with keratitis and catheter-related bacteraemia, and conjunctivitis, respectively (Teng et al, 2015). These suggest that members of the genus Tsukamurella should not be dismissed as harmless in clinical laboratories.…”

“…Self-hybridization values were considered to be 100 %, representing the maximal achievable signal. Values obtained with other test strains were compared with this standard (Teng et al, 2015). Strains HKU54 T and HKU55 possessed 92.1±7.9 % (mean±SD, HKU54 T as probe) or 92.6±3.2 % (mean±SD, HKU55 as probe) DNA-DNA relatedness, suggesting they should belong to the same species.…”

“…Scanning electron microscopy (SEM) was performed according to our previous studies (Woo et al, 2003;Teng et al, 2015) with slight modifications. Bacterial cells were harvested from strains HKU54 T and HKU55 grown on Columbia agar with 5 % defibrinated sheep blood, followed by washing with 1Â PBS and low-speed centrifugation (2000 r.p.m.).…”

“…Preparation of genomic DNA was performed using QIAGEN Genomic tip 100-G (Qiagen). DNA probe was prepared using DIGHigh Prime DNA labelling reagent (Roche Diagnostics) while DNA-DNA dot blot hybridization was carried out using ULTRAhyb ultrasensitive hybridization buffer (Ambion), DIG wash and block buffers, anti-DIG-AP and CDP-Star (Roche Diagnostics) as described previously (Teng et al, 2015). DNA-DNA hybridization was performed at 42 C in triplicate for each of the probes used.…”

“…Bacterial DNA extraction, PCR amplification of the 16S rRNA gene and DNA sequencing were performed according to our previously published protocols (Teng et al, 2015). PCR primers, LPW27807 5¢-TGGCTCAGGACGAACGCT- (Yassin et al, 1996;Kattar et al, 2001;Nam et al, 2003 and from the present study); 8, T. paurometabola ATCC 8368 T (Collins et al, 1988;Kattar et al, 2001;Olson et al, 2007 and from the present study); 9, T. strandjordae ATCC BAA-173 T (Kattar et al, 2001 and from the present study); 10, T. spongiae DSM 44990 3¢ and LPW27808 5¢-GAGGTGATCCAGCCGCA-3¢, were designed targeting the conserved regions of the 16S rRNA gene according to available sequences of species of the genus Tsukamurella.…”