Paracholeplasma manati sp. nov. isolated from Florida manatees (Trichechus manatus latirostris).

Four novel strains of a member of the genus Paracholeplasma (OakleyT, Holly, Lorelei and Ariel) were isolated from skin of Florida manatees (Trichechus manatus latirostris). These strains were phenotypically and genetically characterized and compared with the known species of the genera Acholeplasma (A.), Alteracholeplasma (Al.), Haploplasma (H.), Paracholeplasma (P.) and Mariniplasma (M.). All the strains produced acid from glucose but did not hydrolyze arginine or urea. All were propagated in ambient air supplemented with 5±1 % CO2 at 35-37 °C using SP4-Z, Columbia and brain-heart infusion medium. Colonies on solid medium showed a typical fried-egg appearance and transmission electron microscopy revealed a typical mycoplasma-like cellular morphology. The results of phylogenetic analyses based on partial 16S rRNA, rpoB, gyrB and parE gene sequences and the whole proteome data indicated that the novel species is a unique species but phylogenetically closely related to Paracholeplasma vituli, Paracholeplasma morum and 'Paracholeplasma brassicae'. The average nucleotide identity and digital DNA-DNA hybridization values between strain OakleyT and the closely related species were significantly lower than the accepted thresholds for describing novel prokaryotic species at the genomic level. On the basis of the genomic, phenotypic and phylogenetic properties, the novel strains represent a novel species of the genus Paracholeplasma, for which the name Paracholeplasma manati sp. nov. with the type strain OakleyT (=NCTC 14352T =DSM 110686T) is proposed. The genomic DNA G+C content and complete draft genome size for the type strain are 38.35 % and 1 873 856 bp, respectively.

Neisseria zalophi sp. nov., isolated from oral cavity of California sea lions (Zalophus californianus).

Three independent strains of Neisseria sp. were isolated from the oral cavity of California sea lions (Zalophus californianus) that were admitted to The Marine Mammal Center facilities in California, USA. The strains were isolated from oral swabs by cultivation on Trypticase Soy agar with 5% sheep blood under aerobic conditions. The 16S rRNA gene sequence of these three strains shared 99% similarity, but demonstrated only 97-98% nucleotide similarity to the phylogenetically closest relatives such as N. canis, N. zoodegmatis, N. animaloris, and N. dumasiana. These three strains also shared 99% sequence similarity of their rplF, rpoB, and gyrB gene sequences. Based on the biochemical tests alone (i.e., without genetic analysis of housekeeping genes), it is difficult to discriminate this novel species from N. canis; however, it can be easily discriminated from all phylogenetically closely related species using the sequencing analysis of its housekeeping genes (e.g., rplF, rpoB, or gyrB genes). Thus, genetic testing is indispensable for accurate identification of this species in a routine laboratory practice. The species is an obligate aerobe and able to grow in Mueller-Hinton broth supplemented with 6% NaCl, but the phylogenetically closely related species (N. canis, N. zoodegmatis, N. animaloris, and N. dumasiana) were not. Based on these phenotypic and genotypic characteristics and phylogenetic data, we conclude that these new strains represent a novel species of the genus Neisseria, for which the name Neisseria zalophi sp. nov. is proposed. The type strain is CSL 7565T (= ATCC BAA2455T = DSM 102031T).