Kribbella albertanoniae sp. nov., isolated from a Roman catacomb, and emended description of the genus Kribbella

Abstract

Strain BC640 T was isolated from site CSC13 from a darkgreen biofilm by using the adhesive tape (Fungi Tape DID) technique (Urzì & De Leo, 2001). Growth of colonies (quantified as the number of c.f.u. per cm 2 of adhesive tape) was carried out on R2A medium (Reasoner & Geldreich, 1985) at 28 u C for 15 days. Ten to 20 colonies were selected randomly, preliminarily characterized after transferring to tryptic soy agar (TSA; BBL) and subsequently maintained on yeast extract-malt extract agar [International Streptomyces Project (ISP) medium 2] (Shirling & Gottlieb, 1966).Genomic DNA was extracted as described by Everest & Meyers (2008). The 16S rRNA gene was amplified as described by Cook & Meyers (2003), the gyrB gene as described by Kirby et al. (2010) and the atpD, recA, relA and rpoB genes as described by Curtis & Meyers (2012). Approximately 500 ng template DNA was used in the PCR amplification of the 16S rRNA and gyrB genes, with 1 mg DNA being used for amplification of the atpD, recA, relA and rpoB genes. PCR products were purified using an MSB Spin PCRapace kit (Invitek) and sequenced (Macrogen). Sequence analysis was performed using DNAMAN version Abbreviations: DDH, DNA-DNA hybridization; MLSA, multilocus sequence analysis.The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA, atpD, gyrB, recA, relA and rpoB gene sequences of strain BC640

Description of Kribbella italica sp. nov., isolated from a Roman catacomb

“…Proposed in 1999 (Park et al , 1999), the genus Kribbella contains nocardioform actinobacteria with ll -diaminopimelic acid (DAP) in the cell wall peptidoglycan. The genus description was recently emended by Everest et al (2013). Another recent development associated with the genus was the publication of a multilocus sequence analysis (MLSA) method, which can be used to assess whether an isolate belongs to a novel species without the need for DNA–DNA hybridization (DDH) experiments (Curtis & Meyers, 2012).…”

“…The MLSA study extended previous phylogenetic analysis of strains of the genus Kribbella using the gyrB gene alone (Kirby et al , 2010). With the recent descriptions of Kribbella albertanoniae (Everest et al , 2013), Kribbella endophytica (Kaewkla & Franco, 2013) and Kribbella shirazensis (Mohammadipanah et al , 2013), at the time of writing there are 20 species of the genus Kribbella with validly published names (Parte, 2014). Here we describe the characterization of a novel member of the genus, isolated from the same site as the previously described species K. albertanoniae , Kribbella catacumbae and Kribbella sancticallisti in the Saint Callixstus catacomb in Rome (Everest et al , 2013; Urzì et al , 2008).…”

“…Fig. S1 (available in the online Supplementary Material) shows the sampling locations in the CSC13 cubicle from which the novel species of the genus Kribbella were isolated (Urzì et al , 2008; Everest et al , 2013; present study). Three of the novel species were associated with white/greyish biofilms, while K. albertanoniae BC640 T was isolated from a dark green biofilm.…”

“…Analysis of the isomer of DAP, the whole cell sugar pattern and the phospholipid pattern were carried out as described by Everest et al (2013). Analysis of the respiratory quinones was carried out by the identification service at the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany).…”

“…DDH analysis was performed as a service by the BCCM/LMG Culture Collection as described by Everest et al (2013). Data are displayed as mean DNA–DNA hybridization values with the differences between the means of the reciprocal values given in parentheses.…”