RESUMEN
Seven novel independent strains of Mycoplasma species were isolated from northern elephant seals (ES2806-NAST, ES2806-GENT, ES3157-GEN-MYC and ES3225-GEN-MYC), a harbour porpoise (C264-GENT and C264-NAST), and a California sea lion (CSL7498). These strains were phenotypically and genetically characterized and compared to the known Mycoplasma species. Four strains (C264-GENT, C264-NAST, CSL7498 and ES2806-NAST) hydrolysed arginine but not urea and did not produce acid from carbohydrates. Strains ES2806-GENT, ES3157-GEN-MYC and ES3225-GEN-MYC did not produced acid from carbohydrates and did not hydrolyse arginine or urea; hence, it is assumed that organic acids are used as the energy source for them. All were isolated and propagated in ambient air supplemented with 5±1â% CO2 at +35-37 °C using either SP4 or PPLO medium. Colonies on solid medium showed a typical fried-egg appearance and transmission electron microscopy revealed a typical mycoplasma cellular morphology. The complete genomes were sequenced for all type strains. Average nucleotide and amino acid identity analyses showed that these novel strains were distant from the phylogenetically closely related Mycoplasma species. Based on these data, we propose four novel species of the genus Mycoplasma, for which the name Mycoplasma miroungirhinis sp. nov. is proposed with the type strain ES2806-NAST (=NCTC 14430T=DSM 110945T), Mycoplasma miroungigenitalium sp. nov. is proposed with the type strain ES2806-GENT (=NCTC 14429T=DSM 110944T) and representative strains ES3157-GEN-MYC and ES3225-GEN-MYC, Mycoplasma phocoenae sp. nov. is proposed with the type strain C264-GENT (=NCTC 14344T=DSM 110687T) and Mycoplasma phocoeninasale sp. nov. is proposed with the type strain C264-NAST (=NCTC 14343T=DSM 110688T) and representative strain CSL7498. The genome G+C contents are 24.06, 30.09, 28.49 and 29.05% and the complete genome sizes are 779â550, 815â486, 693â115, and 776â009 bp for strains ES2806-NAST, ES2806-GENT, C264-GENT and C264-NAST, respectively.
Asunto(s)
Mycoplasma , Phocoena , Filogenia , Leones Marinos , Phocidae , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Mycoplasma/clasificación , Mycoplasma/aislamiento & purificación , Phocoena/microbiología , ARN Ribosómico 16S/genética , Leones Marinos/microbiología , Phocidae/microbiología , Análisis de Secuencia de ADNRESUMEN
Two independent strains of a Leptotrichia species (ES3154-GLUT and ES2714_GLU) were isolated from the oral cavity of northern elephant seals (Mirounga angustirostris) that were admitted to The Marine Mammal Centre facilities in California, USA. The strains were isolated from oral swabs by cultivation in PPLO broth supplemented with serum, penicillin and colistin in anaerobic conditions. The strains were Gram-negative, pleomorphic, indole-, oxidase- and catalase-negative, non-spore-forming, non-motile rods/coccobacilli in short chains. The 16S rRNA gene sequence of these strains shared 94.42â% nucleotide similarity with Oceanivirga salmonicida AVG 2115T but demonstrated ≤86.00-92.50â% nucleotide similarity to the 16S rRNA genes of other species of the family Leptotrichiaceae. The genome was sequenced for strain ES3154-GLUT. Average nucleotide identity values between strain ES3154-GLUT and 15 type strain genomes from the family Leptotrichiaceae ranged from 66.74â% vs. Sebaldella termitidis to 73.35â% vs. O. salmonicida. The whole genome phylogeny revealed that the novel species was most closely related to O. salmonicida AVG 2115T. This relationship was also confirmed by nucleotide similarity and multilocus phylogenetic analyses employing various housekeeping genes (partial 23S rRNA, rpoB, rpoC, rpoD, polC, adh, gyrA and gyrB genes). Chemotaxonomic and phenotypical features of strain ES3154-GLUT were in congruence with closely related members of the family Leptotrichiaceae, represented by similar enzyme profiles and fatty acid patterns. MALDI-TOF MS analysis was capable to clearly discriminate strain ES3154-GLUT from all currently described taxa of the family Leptotrichiaceae. Based on these data, we propose a novel species of the genus Oceanivirga, for which the name Oceanivirga miroungae sp. nov. is proposed with the type strain ES3154-GLUT (=DSM 109740T=CCUG 73653T=ATCC TSD-189T=NCTC 14411T) and one representative strain ES2714_GLU. The G+C content is 26.82 %, genome size is 1 356 983 bp.
Asunto(s)
Fusobacterias/clasificación , Boca/microbiología , Filogenia , Phocidae/microbiología , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , California , ADN Bacteriano/genética , Ácidos Grasos/química , Fusobacterias/aislamiento & purificación , Genes Bacterianos , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , ARN Ribosómico 23S , Análisis de Secuencia de ADNRESUMEN
Novel ureaplasma strains have been isolated from the genital tract of both sexes of northern elephant seals (Mirounga angustirostris; six strains) and California sea lions (Zalophus californianus; five strains) stranded along the Central California coast, USA. These strains were phenotypically and genetically characterized and compared to other seven known Ureaplasma species. All novel ureaplasma strains hydrolysed urea, but did not metabolize arginine, and all were isolated and propagated using PPLO medium supplemented with urea under aerobic, microaerophilic, and anaerobic atmospheric conditions at +35-37 °C. Transmission electron microscopy revealed typical mollicute cellular morphology. Molecular characterization included assessment of the following genetic loci: 16S rRNA, the 16S-23S ITS, 23S rRNA, rpoB, ftsH, tufB, rpoC, fusA and ureC. Complete 16S rRNA gene sequence analysis of these novel Ureaplasma species indicated that they were most closely related to each other with nucleotide identity 99.87 % and ≤93.08 % related to other known Ureaplasma species. The results of nucleotide analysis of the sequenced housekeeping genes revealed 71.68-93.02 % similarity to corresponding genes of other known Ureaplasma species. The multi-locus genetic characterization and the phylogenetic analysis of the 16S rRNA and rpoB genes of these Ureaplasma species clearly demonstrated their novelty and, reflecting their host specificites, the name Ureaplasma miroungigenitalium sp. nov. is proposed for the Ureaplasma species isolated from northern elephant seals, the type strain is ES2783-GENT (=DSM 24842T=ATCC BAA-2460T), and the name Ureaplasma zalophigenitalium sp. nov. is proposed for the Ureaplasma species isolated from California sea lions, the type strain is CSL7644-GENT (=DSM 24843T=ATCC BAA-2262T).
Asunto(s)
Genitales/microbiología , Filogenia , Leones Marinos/microbiología , Phocidae/microbiología , Ureaplasma/clasificación , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , Secuencia de Bases , California , ADN Bacteriano/genética , Ácidos Grasos/química , Femenino , Genes Bacterianos , Masculino , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genética , Análisis de Secuencia de ADN , Ureaplasma/aislamiento & purificaciónRESUMEN
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).
Asunto(s)
Boca/microbiología , Neisseria/genética , Leones Marinos/microbiología , Animales , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/análisis , Genotipo , Tipificación Molecular , Neisseria/aislamiento & purificación , Fenotipo , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
Domoic acid (DA) is a naturally occurring neurotoxin known to harm marine animals. DA-producing algal blooms are increasing in size and frequency. Although chronic exposure is known to produce brain lesions, the influence of DA toxicosis on behavior in wild animals is unknown. We showed, in a large sample of wild sea lions, that spatial memory deficits are predicted by the extent of right dorsal hippocampal lesions related to natural exposure to DA and that exposure also disrupts hippocampal-thalamic brain networks. Because sea lions are dynamic foragers that rely on flexible navigation, impaired spatial memory may affect survival in the wild.