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Endolithic metagenome analysis of two beach samples collected form Chilika, Odisha, India indicated rich bacterial diversity. While the metagenome analysis of sample one yielded 16S rRNA gene sequences which represent six phyla and 16 genera, sample two yielded very rich diversity representing 17 phyla and 286 genera. Six species of bacteria were isolated from the endolithic enrichments and most of them have 16S rRNA gene sequence similarity of >99 % with known taxa, except for strain JC140(T) having <97 % sequence similarity with taxa with valid names. Strain JC140(T) is Gram-stain-negative and motile. 16S rRNA gene based EzTaxon-e BLAST analysis indicated that strain JC140(T) is closely related with the type strains of Rhizobium yanglingense SH22623(T) (96.8 %), R. alkalisoli CCBAU 01393(T) (96.3 %), R. vignae CCBAU 05176(T) (96.2 %), R. mesosinicum CCBAU 25010(T) (96.1 %) and other members of the genus Rhizobium with <96 % sequence similarity. Strain JC140(T) was characterized based on polyphasic taxonomic analysis. Strain JC140(T) could tolerate up to 5 % salinity, resistant to rifampicin (10 μg) and was positive for catalase and oxidase. The major fatty acid is summed feature 8 (C18:1ω7c/C18:1ω6c) with minor amounts of C19:0 cycloω8c, C16:0, C18:0, C17:0cyclo, summed feature 2 (C14:03OH/C16:1 isoI), summed feature 3 (C16:1ω7c/C16:1ω6c) and 2-hydroxyacid of C15:0. Diphosphatidylglycerol, phosphatidylethanol-amine, phosphatidylcholine, phosphatidylglycerol, unidentified lipids (L1, 2), unidentified phospholipid (PL1-4) constitute the polar lipids of strain JC140(T). The strain has bacteriohopane derivative (BHD2), diplopterol and two unidentified hopanes (UH1, 2) as major hopanoids. Q-10 is the major quinone of strain JC140(T). Based on polyphasic taxonomic analysis, strain JC140(T) represents a species in the genus Rhizobium for which, the name R. endolithicum sp. nov. is proposed. The type strain is JC140(T) (= KCTC32077(T) = CCUG64352(T) = MTCC11723(T) = HAMBI 2447(T)).

Purpose Previous studies indicate that the polymerase chain reaction (PCR) nasal assay for methicillin-resistant Staphylococcus aureus (MRSA) has a consistently high (&gt;95%) negative predictive value (NPV) in ruling out MRSA pneumonia; however, optimal timing of PCR assay specimen and respiratory culture collection is unclear. Methods A study including 736 patients from a community hospital system was conducted. Patients were included if they had undergone MRSA nasal screening with a PCR assay and had documented positive respiratory culture results. Results In the full cohort, the MRSA PCR nasal screen assay was demonstrated to have an NPV of 94.9% (95% confidence interval [CI], 92.8%-96.5%) in ruling out MRSA-positive respiratory cultures. When evaluating the NPV by level of care (ie, where the MRSA PCR nasal assay sample was collected), no significant difference between values for samples collected in an intensive care unit vs medical/surgical units was identified (NPV [95%CI], 94.9% [92.7%-96.6%] vs 95.3% [88.4%-98.7%]).Additionally, NPV remained high with use of both invasive (NPV [95%CI], 96.8% [92.7%-99.0%]) and noninvasive (NPV [95%CI], 94.5% [91.7%-96.2%]) respiratory sampling methods. Finally, when evaluating the effect of time between MRSA PCR nasal screening and respiratory sample collection, we found high NPVs for all evaluated timeframes: within 24 hours, 93.8% (90.1%-96.4%); within 25 to 48 hours, 98.6% (92.7%-100.0%); within 49 hours to 7 days, 95.7% (91.4%-98.3%); within 8 to 14 days, 92.9% (85.1%-97.3%); and after more than 14 days, 95.5% (84.5%-99.4%). Conclusion We report high NPVs for up to 2 weeks between specimen collections, which allows clinicians to use a negative MRSA PCR nasal screen assay to rule out MRSA pneumonia, potentially leading to decreased exposure to MRSA-active antibiotics.

Rhizobium subbaraonis sp. nov., an endolithic bacterium isolated from beach sand

Strain JC268T was isolated from pebbles collected from a dam located in Lalitpur, Uttar Pradesh, India. Cells of strain JC268T were coccoid, appeared in pairs/triads/tetrads or short chains and were Gram-stain-positive, non-spore-forming, non-motile and obligately aerobic. Strain JC268T was catalase- and oxidase-positive and utilized citrate for growth. The genomic DNA G+C content of strain JC268T was 65.3 mol%. The cell-wall peptidoglycan contained l-lysine–l-serine–d-aspartic acid as interpeptide bridge with the type A4α. The major menaquinone was MK-8(H4). Major (>10 %) fatty acids were iso-C16 : 0, iso-C16 : 1H and anteiso-C17 : 1ω9c. Diphosphatidylglycerol, phosphoglycolipid, phosphatidylinositol, glycolipid, four unidentified lipids, an amino lipid and phospholipid were the polar lipids of strain JC268T. EzTaxon-e blast search of 16S rRNA gene sequences showed that strain JC268T has highest similarity to Barrientosiimonas humi 39T (98.65 %) and Tamlicoccus marinus MSW-24T (97.8 %) of the family Dermacoccaceae. Genome reassociation (based on DNA–DNA hybridization) of strain JC268T with Barrientosiimonas humi CGMCC 4.6864T ( = 39T) and T. marinus KCTC 19485T ( = MSW-24T) yielded values of 32.5 ± 2 % and 27.3 ± 2 %, respectively. Based on the data from phylogenetic and polyphasic taxonomic analyses, strain JC268T represents a novel species of the genus Barrientosiimonas for which the name Barrientosiimonas endolithica sp. nov., is proposed. The type strain of Barrientosiimonas endolithica is JC268T ( = KCTC 29672T = NBRC 110608T). Our data suggest that T. marinus should be reclassified within the genus Barrientosiimonas. Thus, a reclassification is proposed for T. marinus, the type and only species of the genus Tamlicoccus, as Barrientosiimonas marina comb. nov., which implies the emendation of the description of the genus Barrientosiimonas.

Strain JC267T was isolated from pebbles collected from Pingleshwar beach, Gujarat, India.Cells are Gram-stain-positive, facultatively anaerobic, non-motile rods forming sub-terminal endospores in swollen ellipsoidal to oval sporangia. Strain JC267 T contains anteiso-C 15 : 0 , iso-C 15 : 0 , iso-C 14 : 0 , iso-C 16 : 0, C 16 : 0 and anteiso-C 17 : 0 as major (.5 %) cellular fatty acids. Polar lipids include phosphatidylglycerol, phospholipids (PL1-3), glycolipids (GL1-2) and an unidentified lipid. Cell-wall amino acids are composed of diagnostic mesodiaminopimelic acid, DL-alanine and a small amount of D-glutamic acid. The genomic DNA G+C content of strain JC267 T is 45.5 mol%. The 16S rRNA gene sequence of strain JC267T showed highest sequence similarities of ,98.41 % with all species of the genus Bacillus when subjected to EzTaxon-e BLAST analysis. The reassociation values based on DNA-DNA hybridization of strain JC267 T with Bacillus halosaccharovorans IBRC-M 10095 T and Bacillus niabensis JCM 16399 T were 26¡1 % and 34¡3 %, respectively. Based on taxonomic data obtained using a polyphasic approach, strain JC267 T represents a novel species of the genus

BackgroundThe MRSA nasal PCR assay is a rapid, noninvasive test that has demonstrated a strong negative predictive value (NPV), as high as 99%, for ruling out MRSA pneumonia. These findings are based primarily on literature from large academic centers, which have evaluated both the positive predictive value (PPV) and NPV of MRSA nasal PCR assays. Investigators sought to assess the NPV of the MRSA nasal PCR assay to rule out MRSA pneumonia within a community healthcare system. To the best of our knowledge, this is the largest study from a community hospital and the only study from a community healthcare system for the utilization of a nasal PCR assay to rule out MRSA pneumonia.MethodsThis is a multicenter, retrospective study of adult patients with both an MRSA nasal PCR assay and positive respiratory culture (sputum, bronchoalveolar lavage, or endotracheal aspirate). Data were collected from September 2014 through August 2015 at three community hospitals (bed size ranging from 328 to 706) across two states within a healthcare system. The study was approved by the Baptist Memorial Hospital Institutional Review Board. PPV and NPV 95% confidence intervals (95% CI) were calculated as previously described in the literature.ResultsA total of 808 patients were included in the analysis across the three hospitals. The total incidence of MRSA in positive sputum samples was 14.9% across the three facilities. Our study demonstrated an overall NPV of 95.1% (93.2, 96.6%) and a PPV of 65.9% (95% CI 57.2, 73.9%). The high NPV was retained despite unit type, resulting in 94.9% (95% CI 92.7, 96.6%), 96.3% (95% CI 90.8, 99.0%), and 94.7% (95% CI 74.0, 99.9%) for the intensive care units (ICU), medical-surgical units, and the emergency department, respectively (Table 1).ConclusionWe concluded that the high NPV of a negative MRSA nasal PCR assay to rule out MRSA pneumonia persisted within a community hospital system. With the results of our study, we plan to utilize institution-specific data along with previously published literature to encourage earlier discontinuation of anti-MRSA antibiotics in patients being treated for pneumonia with negative MRSA nasal PCR assays. Our study demonstrates the validity of the assay in the large community hospital setting with similar findings to studies at large academic institutions. Disclosures All authors: No reported disclosures.