Rare Spiroplasma Bloodstream Infection in Patient after Surgery, China, 2022.

We report a case of Spiroplasma bloodstream infection in a patient in China who developed pulmonary infection, acute respiratory distress syndrome, sepsis, and septic shock after emergency surgery for type A aortic dissection. One organism closely related to Spiroplasma eriocheiris was isolated from blood culture and identified by whole-genome sequencing.

Preliminary comparative genomics analysis among Corynebacterium kroppenstedtii complex necessitates a reassessment of precise species associated with mastitis.

AIMS: This study aimed to characterize the first complete genome of Corynebacterium parakroppenstedtii and clarify the evolutionary relationship in the Corynebacterium kroppenstedtii complex (CKC) by using comparative genomics analysis. METHODS AND RESULTS: The genome of isolate yu01 from a breast specimen was sequenced, and 35 CKC genomes were collected. Analysis of 16S rRNA, rpoB, and fusA suggested ambiguous identification, whereas ANI analysis assigned isolate yu01 as Coryne. parakroppenstedtii. The fourth genospecies "Corynebacterium aliikroppenstedtii" was identified in CKC. Comparative genomics analysis suggested that the genomic arrangement in CKC was highly conserved. A total of 43 potential virulence genes and 79 species-specific genes were detected. Most genome-based phylogenetic analysis were incapable of resolving the interspecific evolutionary relationships among CKCs. A total of 20 core genes were found to be distinguishable in CKC. CONCLUSIONS: This study suggested the limited divergence and unavailability of normal single gene-based identification in CKC and questioned the precise species of strains associated with mastitis, identified as Coryne. kroppenstedtii in previous studies. The 20 genes showed potential to enhance the methods for the identification and epidemiological investigation of CKC.

Infection may play an important role in the pathogenesis of alveolar osteonecrosis following facial herpes zoster: a case report and literature review.

BACKGROUND: Herpes zoster (HZ) is one of the most common skin diseases caused by viruses. Facial HZ develops when the varicella-zoster virus affects the trigeminal nerve, and alveolar osteonecrosis is a rare complication. However, the exact pathogenesis of postherpetic alveolar osteonecrosis remains unclear. CASE DESCRIPTION: We encountered a patient who presented to the dermatology clinic with facial HZ and tooth exfoliation in the upper right jaw, and panoramic radiography revealed decreased bone density and poor alveolar socket healing in his right maxilla. Biopsy of the alveolar process revealed fragments of nonvital lamellar bone, which were devoid of osteoblasts and osteocytes and were surrounded by numerous neutrophils and bacterial aggregates. Thus, the diagnosis of alveolar osteonecrosis following facial HZ was confirmed. He then underwent resection of the osteonecrotic tissue. The pathological findings of postoperative tissue were similar to those of previous biopsies. Varicella-zoster virus and multiple types of bacteria were detected through next-generation sequencing, and the species of bacteria were consistent with the results of bacterial culture. Antibiotics and valaciclovir were administered during the perioperative period. The patient showed good recovery at the 9-month follow-up. CONCLUSIONS: The coexistence of bacterial and viral infection may play an important role in the pathogenesis of alveolar osteonecrosis following HZ. To our knowledge, we are the first to directly explore microbial pathogens in a case of postherpetic alveolar osteonecrosis through next-generation sequencing and bacterial culture. We recommend that oral examinations be carefully conducted for patients who are diagnosed with facial HZ, even if their facial rashes have faded away. We suggest that a prolonged and full-dose antiviral therapy course may be beneficial for the treatment of facial HZ with intraoral lesions. The implementation of dental preventive measures should be considered for patients with facial HZ. The application of antibiotics and excision of necrotic bone may reduce the abundance of bacteria in lesions and improve wound healing.

Curtanaerobium respiraculi gen. nov., sp. nov., a novel anaerobic bacterium isolated from human bronchoalveolar lavage fluid.

Two related anaerobic strains, designated as SWB101512T and SWB19611, were isolated from the bronchoalveolar lavage fluid of two lung cancer patients. Cells were Gram-stain-positive, non-motile and non-spore-forming. Growth could be observed at 26-45 °C (optimum, 37 °C), pH 5.0-8.5 (optimum, pH 7.0) and with 0.5-2.0 % (v/w) NaCl (optimum, 1.0%). The 16S rRNA gene sequences of SWB101512T and SWB19611 showed the highest similarities to Denitrobacterium detoxificans DSM 21843T (91.1 and 91.3 %, respectively). The phylogenetic tree based on the 16S rRNA gene sequences and the core genome sequences demonstrated that the two strains clustered together and formed a distinct lineage within the family Eggerthellaceae. The DNA G+C contents of strains SWB101512T and SWB19611 were 62.0 and 61.9 mol%, respectively. The predominant cellular fatty acids of strains SWB101512T and SWB19611 were C16 : 0 DMA (27.8 and 28.8 %, respectively). The respiratory menaquinone in both strains was menaquinone 6 and the polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, two phospholipids, three glycolipids and three unidentified lipids. Based on evidence from phenotypic, chemotaxonomic and genomic analyses, a new genus and species belonging to the family Eggerthellaceae, named Curtanaerobium respiraculi gen. nov., sp. nov. is proposed. The type strain is SWB101512T (=GDMCC 1.2991T=JCM 35330T).

Fluviispira vulneris sp. nov., isolated from human wound secretions.

Human infections by environmental bacteria is becoming an increasing problem and has become a matter of great concern due to the adverse effects worldwide. In this study, we reported a new environmental pathogen. Isolate GX5518T was a novel Gram-negative, aerobic, non-motile, pleomorphic and red-pigmented bacterium, was isolated from human wound secretions (GuangXi, People's Republic of China). Growth occurred at pH 6.0-8.0 (optimum, pH 7.0) and 10-37 °C (optimum, 28-32 °C) with 0-1.5% (w/v) NaCl in R2A agar. Comparative analysis of the 16S rRNA gene sequences revealed that isolate GX5518T was closely related to Fluviispira sanaruensis JCM 31447T (99.73%) and Fluviispira multicolorata 33A1-SZDPT (98.49%). However, the estimated ANI values of the isolate GX5518T compared to the F. sanaruensis JCM 31447T and F. multicolorata 33A1-SZDPT were 88.67% and 77.35%, respectively. The estimated dDDH, ANI and AAI values between isolate GX5518T and its closely related strains were below the threshold values generally considered for recognizing a new species. The genome size was 3.6 Mbp and the DNA G + C content was 33.1%. The predominant fatty acids (> 5%) in GX5518T cells were iso-C15:0, C16:0, C17:0, C17:1 ω8c and C16:1 ω7c/C16:1 ω6c. The major menaquinone was MK-8 (86.9%). The polar lipids were phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and three unknown lipids (L1-3). The chemical composition was different from that of the F. sanaruensis JCM 31447T. Comparative genomics analysis between isolate GX5518T and its related strains revealed that there were a number of genes involved in resistance to antibiotics and toxic compounds in isolate GX5518T, which were responsible for the copper homeostasis, cobalt-zinc-cadmium resistance, resistance to fluoroquinolones, and zinc resistance. Based on the phenotypic, chemotaxonomic, and genomic analyses, isolate GX5518T (= CGMCC 1.18685T = KCTC 82149T) represents a novel species of the genus Fluviispira, for which the name Fluviispira vulneris sp. nov. is proposed.

Corynebacterium lipophilum sp. nov., a lipophilic bacterium isolated from clinical breast specimens and emended description of the species Corynebacterium pilbarense.

Two isolates (MC-18T and MC-17D), representing the Gram-stain-positive, facultatively anaerobic, irregular rod-shaped, non-motile, and non-spore-forming actinobacteria, were isolated from clinical breast specimens in Guangzhou, China. The growth of the isolates is enhanced by supplementing 1% Tween-80 on Luria Bertani agar. Optimal growth of the isolates was observed at 37 °C, pH 7-8, and with 1% (w/v) NaCl on Columbia blood agar. Pairwise comparison of the 16S rRNA gene sequences revealed that isolates MC-18T and MC-17D shared the highest sequence similarities with Corynebacterium liangguodongii 2184T (96.9%), which were lower than the threshold value for species delineation (98.65%). Phylogenetic dendrograms based on the 16S rRNA gene, rpoB gene, and core genomes indicated that two isolates formed a distinct lineage within the genus Corynebacterium. The estimated dDDH, ANIb, ANIm, and AAI values between strain MC-18T and its closely related strains were below the threshold values generally considered for recognizing a new species. The genome DNA G + C contents of both the isolates MC-18T and MC-17D are 60.6%. The two isolates have virulence-related genes of the VF classes of adhesion and antiphagocytosis, and also contain the antimicrobial resistance genes ErmX, mtrA, rpoB2, and RbpA. The major fatty acids (> 10%) of isolates MC-18T and MC-17D were C16:0, C18:1 ω9c, C18:0 and summed feature 5 (anteiso-C18:0 and/or C18:2 ω6,9c). The main respiratory quinone of strain MC-18T was MK-8(H2), and the polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, three unidentified glycolipids, an unidentified aminolipid, and four unidentified phosphoglycolipids. The two isolates lack mycolic acids in the cell envelope. Based on the above findings, the two isolates are considered to represent a novel species of the genus Corynebacterium, for which the name Corynebacterium lipophilum sp. nov. is proposed, with isolate MC-18T (= NBRC 115144T = CCTCC AB 2020201T) as the type strain. An emended description of the Corynebacterium pilbarense is also provided.

Copranaerobaculum intestinale gen. nov., sp. nov., a novel anaerobic bacterium isolated from human faeces.

A novel obligate anaerobic organism, designated DONG20-135T, was isolated from human faeces collected in Beijing, PR China. Cells were Gram-stain-negative, rod-shaped, non-motile and non-spore-forming. Growth occurred at 25‒45 °C (optimum, 30‒35 °C), a pH range of 6-9 (optimum, pH 8) and in the presence of 0‒3.5 % (w/v) NaCl (optimum, 0.5‒1.5 %). The major fatty acids were C16 : 0, C18 : 1 ω9c and C10 : 0, the polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, four glycolipids, six aminolipids, three aminophospholipids and four unidentified lipids. No respiratory quinones were detected. The cell-wall peptidoglycan of the strain was A1γ type, containing meso-diaminopimelic acid. The 16S rRNA gene sequences shared a lower identity (<92.7 % similarity) with the described species. The phylogenetic tree based on 16S rRNA gene sequences and the protein-concatamer tree showed that strain DONG20-135T formed a distinct lineage within the family Erysipelotrichaceae. The genomic DNA G + C content was 42.2 mol%. Based on the results of phenotypic, chemotaxonomic and genomic analyses, strain DONG20-135T represents a novel genus of the family Erysipelotrichaceae, for which the name Copranaerobaculum intestinale gen. nov., sp. nov. is proposed (=KCTC 15868T=CGMCC 1.17357T).

Characterization of carbapenem-resistant Acinetobacter baumannii ST540 and Klebsiella pneumoniae ST2237 isolates in a pneumonia case from China.

AIMS: This study aimed to characterize the chromosome and plasmid sequences, and determine the transferability of plasmids in carbapenem-resistance Acinetobacter baumannii DD520 and Klebsiella pneumoniae DD521 isolates from the same patient who was co-infected in a hospital in China. METHODS AND RESULTS: Both isolates DD520 and DD521 exhibited multidrug resistance phenotype, especially the former isolate which was resistant to nine classes of antimicrobials including carbapenems, quinolones, penicillins, cephalosporins, tetracyclines, phenicols, fosfomycins, sulfanilamides and aminoglycosides. Carbapenem resistance genes of blaOXA-23 and blaOXA-66 were identified on the chromosome of A. baumannii DD520, and blaKPC-2 was found in the plasmid pDD521.2 from K. pneumoniae DD521. Phylogenetic analysis revealed that A. baumannii DD520 belonged to the ST540 clone, and K. pneumoniae DD521 belonged to the ST2237 clone. Plasmid analysis suggested that blaKPC-2 was embedded into plasmid pDD521.2, which might be resulted from IS26- and Tn1721-mediated transposition. Plasmid pDD521.2 carrying blaKPC-2 successfully transferred from K. pneumoniae DD521 into Escherichia coli C600, and carbapenems resistance also transferred in the conjugation. CONCLUSIONS: To our knowledge, it was the first report of A. baumannii ST540 and K. pneumoniae ST2237 in the same patient in China. Both these two isolates exhibited resistance to carbapenem, which was likely to have resulted from carbapenem-resistance genes blaOXA-23 -blaOXA-66 on the chromosome of A. baumannii ST540, and blaKPC-2 in the plasmid of K. pneumoniae ST2237. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study highlighted that effective measures were urgent to prevent and control the co-infection caused by two or more carbapenem-resistance pathogens in the same patient.

Sandaracinobacteroides hominis gen. nov., sp. nov., isolated from human skin.

Strain SZY PN-1 T, representing a novel Gram-negative, aerobic, non-motile, rod-shaped and yellow-pigmented bacterium, was isolated from a skin sample of a healthy Chinese male. Growth occurred at pH 6.0-8.0 (optimum, pH 7.0) and 10-37 â„ƒ (optimum, 30 â„ƒ) with 0-1.0% (w/v) NaCl in R2A agar. Comparative analysis of the 16S rRNA gene sequences revealed that strain SZY PN-1 T shared high similarities with two invalid-published species, "Sandaracinobacter sibiricus" RB16-17 (97.1%) and "Sandaracinobacter neustonicus" JCM 30734 (96.6%), respectively. Phylogenetic analysis of 16S rRNA gene sequences together with protein-concatemer tree showed that SZY PN-1 T formed a separate branch within the family Sphingosinicellaceae. The DNA G + C content of the strain SZY PN-1 T was 65.0% (genome). The polar lipid profile included phosphatidylethanolamine, phosphatidylglycerol, two sphingoglycolipids, diphosphatidylglycerol, five unidentified glycolipids, and seven unidentified lipids. The predominant fatty acids (> 10.0%) were identified as C18:1 ω7c and/or C18:1 ω6c, C17:1 ω6c, C16:1 ω7c and/or C16:1 ω6c. The major respiratory quinone was Q-10. Based on the phenotypic and genotypic features, a novel genus and species, Sandaracinobacteroides hominis gen. nov., sp. nov. is proposed, with type strain SZY PN-1 T (= KCTC 82150 T = NBRC 114675 T).

Legionella septentrionalis sp. nov., isolated from aquatic environments in the northern PR China.

Four strains (km711T, km714, km542 and km524), representing a novel Legionella species, were isolated from aquatic environments in northern PR China. Cells were Gram-stain-negative, rod-shaped, microaerobic, motile and growth depended on l-cysteine. They grew at 25‒42 °C (optimum, 35‒37 °C) and could tolerate up to 1.5 % (w/v) NaCl (optimum, 0.5 %). The major fatty acids (>5 %) of the type strain km711T were C17 : 0 anteiso, C15 : 0 anteiso, iso-C16 : 0 and C16 : 1 ω7c and/or iso-C15 : 0 2OH. The pairwise comparison values were <96.1 % for 16S rRNA gene sequences, 23.3‒28.7 % interspecies variation for mip gene sequences, <93.6 % average nucleotide identity and <72.8 % average amino acid identity between these four strains and related type strains within the genus Legionella. The phylogenetic tree based on the four concatenated genes (16S rRNA, mip, rpoB and rnpB) and protein-concatamer tree based on concatenation of 21 protein markers both revealed that these four strains formed a separate phylogenetic branch cluster within the genus Legionella. The results of phenotypic and genotypic features suggest that these four strains represent a novel species of the genus Legionella, for which the name Legionella septentrionalis sp. nov. is proposed (type strain km711T=KCTC 15655T=NBRC 113219T).

Cysteiniphilum marinum sp. nov., isolated from coastal seawater.

Six aerobic Gram-negative bacteria were isolated from seawater in Guangdong Province, P.R. China. Cells were observed to be Gram-negative, aerobic, non-motile and non-spore forming. Growth of the designated type strain 19X3-30T occurred at a temperature range of 14-37 °C (optimum, 28 °C), a pH range of 6.0-8.0 (optimum, pH 7) and up to 7.5% NaCl (optimum, 1.5%; w/v), and was enhanced by CO2 and L-cysteine supplementation. The major polar lipids identified in strain 19X3-30T were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The principal cellular fatty acids profile showed the presence of anteiso-C15:0, anteiso-C17:0 and C18:0 (> 8% of total fatty acids), and the respiratory quinone was ubiquinone 8 (UQ-8). According to the analysis of 16S rRNA gene sequences, these strains represented a novel species within the family Fastidiosibacteraceae, sharing maximum similarities with Cysteiniphilum litorale DSM 101832T (96.6%) and Cysteiniphilum halobium DSM 103992T (95.3%). Phylogenetic dendrograms based on 16S rRNA gene and protein marker genes from the genomic sequences both indicated that the strains formed a monophyletic lineage closely linked to the genus Cysteiniphilum, which was also supported by the UPGMA dendrogram based on the MALDI-TOF MS profile. The genomic DNA G + C contents of six strains ranged from 38.0% to 38.1%. Based on different taxonomic genomic metrics, phylogeny and phenotypic features, we propose that the strains warrant the assignment to a novel species, for which the name Cysteiniphilum marinum sp. nov. is proposed. The type strain is 19X3-30T (= KCTC 82154T = CGMCC 1.18585T).

Pigmentibacter ruber gen. nov., sp. nov., a novel bacterium of the family Silvanigrellaceae isolated from human blood.

A Gram-negative, aerobic, non-motile, pleomorphic, red-pigmented bacterium, designated HNSRY-1T, was isolated from the blood sample of a near drowning patient in Republic of China. Strain HNSRY-1T grew at 15-37 °C (optimum, 35 °C), with pH 6.0-8.0 (optimum, pH 7.0) and 0-1.5% (W/V) NaCl (optimum, 1%). The predominant fatty acids (> 5%) in HNSRY-1T cells are iso-C15:0, C17:0, C17:1 ω8c, C16:0, and C16:1 ω6c/C16:1 ω7c. The major respiratory quinone is MK-8. The polar lipids are phosphatidylglycerol, phosphatidylethanolamine, three unidentified lipids and four unidentified aminolipids. The 16S rRNA gene sequence-based phylogenetic analysis indicated that strain HNSRY-1T belonged to the family Silvanigrellaceae, forming a distinct phylogenetic line distantly related (< 96.4% sequence similarity) to known species of the family. The ANI values of strain HNSRY-1T compared to the closely related species were below the determined genus division threshold limit (92-94% ANI), and AAI values were lower than the determined genus division threshold limit (80% AAI). Whole genome sequencing revealed a genome size of 3.63 Mb with a DNA G + C content at 29.6%. The half-lethal dose of strain HNSRY-1T on KM mice is about 1.12 × 108 CFU/ml. Virulence gene analysis showed that the pathogenicity of HNSRY-1T may be related to tufA, htpB, katA, wbtL, wbtM, pseB, clpP, cheY, cheV3, acpXL, pilB, fliN, ggt, flgG, fliP, nueB, pseA, bioB and flil. Based on these findings from the polyphasic taxonomy studies, a novel genus and species of the family Silvanigrellaceae. Pigmentibacter ruber gen. nov., sp. nov. is proposed, with type strain HNSRY-1T (= KCTC 72920T = CGMCC 1.18525T).

Haemophilus seminalis sp. nov., isolated from human semen.

Two Haemophilus-like isolates with similar biochemical characteristics, designated strains SZY H1T and SZY H2, were isolated from human semen specimens. Cells were Gram-negative, non-motile, non-acid-fast, pleomorphic rods or coccobacilli. The major fatty acids (>10 %) were C16 : 0, C14 : 0, iso-C16 : 0 and/or C14 : 0 3-OH and C16 : 1 ω6c and/or C16 : 1 ω7c. The polar lipids were determined to be phosphatidylethanolamine, phosphatidylglycerol, an unidentified phospholipid, an unidentified aminophospholipid, two unidentified polar lipids and four unidentified aminolipids. The major polyamine was found to be cadaverine. The near-full-length (1462 nt) 16S rRNA gene sequences analysis showed the two isolates were nearly identical (>99.8 %), and closely matched Haemophilus haemolyticus ATCC 33390T with 98.9-99.1 % sequence similarities. Phylogenetic analysis based on 16S rRNA gene sequences and concatenation of 30 protein markers also revealed that the isolates clustered together with H. haemolyticus ATCC 33390T, and formed a distinct lineage well separated from the other members of the genus Haemophilus. Further, the average nucleotide identity values between the two isolates and their related species were below the established cut-off values for species delineation (95 %). Based on these findings, the two isolates are considered to represent a new species of the genus Haemophilus, for which name Haemophilus seminalis sp. nov. is proposed. The type strain is SZY H1T (=NBRC 113782T=CGMCC 1.17137T).

Francisella salimarina sp. nov., isolated from coastal seawater.

Four strains (SYSU SYW-1T, SYW-2, SYW-3 and XLW-1) were isolated from seawater near the shore in Guangdong Province, China. Cells were Gram-stain-negative, aerobic, non-motile and non-spore-forming. Growth was observed at a temperature range of 16-40 °C (optimum, 32 °C), a pH range of 4-8 (optimum, pH 7) and in the presence of up to 10 % (w/v) NaCl. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and an unidentified phospholipid. The respiratory quinone was ubiquinone 8 (UQ-8), and the predominant fatty acids were C18 : 0 3-OH, C10 : 0, C14 : 0 and C18 : 1ω9c. Comparison of 16S rRNA gene and genome sequences confirmed that these strains represented a novel member of the genus Francisella, with less than 98.8 % 16S rRNA gene sequence similarity and less than 95 % genomic average nucleotide identity to recognized Francisella species. The phylogenetic tree based on 16S rRNA gene sequences and the protein-concatamer tree based on a concatenation of 28 protein marker sequences both indicated that the strains clustered with 'Francisella salina' TX07-7308 and 'Francisella marina' E95-16, but formed a distinct lineage group among the other members of the genus Francisella. The DNA G+C contents of the four strains were determined to be 32.9, 32.7, 32.9 and 32.9 %, respectively (genome). On the basis of phenotypic and genotypic features, the strains are considered to represent a novel species of the genus Francisella, for which the name Francisella salimarina sp. nov. is proposed. The type strain is SYSU SYW-1T (=CGMCC 1.17031T=NBRC 113781T).

Vogesella urethralis sp. nov., isolated from human urine, and emended descriptions of Vogesella perlucida and Vogesella mureinivorans.

A novel Vogesella strain, YM-1T, was recovered from human urine in PR China in 2017. Cells of strain YM-1T were Gram-stain-negative, rod-shaped, aerobic, motile, non-spore-forming and poly-ß-hydroxybutyrate-accumulating. The strain contained C16:1ω6c/C 16:1ω7c, C16:0 and C18:0ω7c as major fatty acids; phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified phospholipid as major polar lipids; and ubiquinone-8 as the predominant respiratory quinone. Comparison of 16S rRNA gene sequences indicated that this strain had highest similarities to Vogesella perlucida DS-28T (98.8 %) and Vogesella mureinivorans 389T (98.1 %). The results of phylogenetic analysis based on the 16S rRNA gene sequences revealed that the novel strain was clustered and well separated with V. perlucida DS-28T and V. mureinivorans 389T within the genus Vogesella. The average nucleotide identity (ANI) and amino acid identity (AAI) analyses showed that this strain was not identified as V. perlucida DS-28T or V. mureinivorans 389T, with values well below the threshold limit for species demarcation (ANI <88.1 %, AAI <88.6 %). Based on the above results, strain YM-1T is proposed to be a novel species of the genus Vogesella with the name Vogesella urethralis sp. nov. (YM-1T=NBRC 113779=CGMCC 1.17135).

Antibiotic Resistance and Molecular Epidemiological Characteristics of Streptococcus agalactiae Isolated from Pregnant Women in Guangzhou, South China.

Streptococcus agalactiae colonization in pregnant women can cause postpartum intrauterine infections and life-threatening neonatal infections. To formulate strategies for the prevention and treatment of S. agalactiae infections, we performed a comprehensive analysis of antibiotic resistance and a molecular-based epidemiological investigation of S. agalactiae in this study. Seventy-two S. agalactiae strains, collected from pregnant women, were subjected to antibiotic susceptibility tests; then, the screened erythromycin and clindamycin nonsusceptible isolates were used for macrolides and clindamycin resistance genes detection, respectively. Detection of resistance genes, serotyping, and determination of virulence genes were performed by polymerase chain reaction. The clonal relationships among the colonized strains were evaluated by multilocus sequence typing. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) mass peak analysis was performed to discriminate the specific sequence types (STs). In our study, 69.4% and 47.2% of the strains were nonsusceptible to erythromycin and clindamycin, respectively; the multidrug resistance rate was 66.7%. All erythromycin nonsusceptible strains harbored resistance genes, whereas only 52.9% of the clindamycin nonsusceptible strains possessed the linB gene. Erythromycin resistance was mainly mediated by the ermB or mefA/E genes. Four serotypes were identified, and the most common serotype was serotype III (52.8%), followed by Ib (22.2%), Ia (18.0%), and II (4.2%). All the strains were divided into 18 STs that were assigned to nine clonal complexes. Most of the major STs were distributed into specific serotypes, including ST19/serotype III, ST17/serotype III, ST485/serotype Ia, ST862/serotype III, and ST651/serotype III. Analysis of virulence genes yielded seven clusters, of which bca-cfb-scpB-lmb (61.6%) was the predominant virulence gene cluster. Among all ST strains distributed in this region, only the ST17 strains had a mass peak at 7620 Da. The outcomes of this study are beneficial for the epidemiological comparison of colonized S. agalactiae in different regions and may be helpful for developing the strategies for the prevention of S. agalactiae infection in Guangzhou. Furthermore, our results show that MALDI-TOF MS can be used for the rapid identification of the ST17 strains.

Facilibium subflavum gen. nov., sp. nov. and Cysteiniphilum halobium sp. nov., new members of the family Fastidiosibacteraceae isolated from coastal seawater.

Two Francisella-like bacteria, designated strains SYSU SYW-5T and SYSU SYW-6T, were isolated from coastal seawater sampled at Huizhou Double-Moon Bay, Guangdong Province, PR China. Cells were found to be Gram-stain-negative, non-motile, non-spore-forming and of pleomorphic shape (coccobacilli or rod). Chemotaxonomic analysis of the plasma membrane revealed ubiquinone-8 as the respiratory quinone, and saturated branched-chain (anteiso-C15 : 0) and saturated straight-chain (C18 : 0) fatty acids as major components (>8 % of total fatty acid). The major polar lipids comprised diphosphatidylglycerol (cardiolipin), phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine, as well as an unidentified aminophospholipid and an unidentified phospholipid in SYSU SYW-5T, and two unknown polar lipids in SYSU SYW-6T. Comparison of 16S rRNA gene sequences showed that SYSU SYW-5T had maximum similarity to Fastidiosibacter lacustris SYSU HZH-2T (93.4 %), while SYSU SYW-6Thad maximum similarity to Cysteiniphilum litorale SYSU D3-2T (97.5 %). Phylogenetic dendrograms based on 16S rRNA genes and 31 concatenated protein markers revealed that the two novel strains represent a distinct lineage within the family Fastidiosibacteraceae. The average nucleotide identity and in silico DNA-DNA hybridization values between the two isolates and their related species were well below the threshold limit for species delineation (<89.2 and <35 %, respectively). Based on the above results, strain SYSU SYW-5T is proposed to be a novel species of a new genus with the name Facilibium subflavum gen. nov., sp. nov. (SYSU SYW-5T = DSM 103991T= KCTC 52556T), and strain SYSU SYW-6T is proposed to be a novel species of genus Cysteiniphilum with the name Cysteiniphilumhalobium sp. nov. (SYSU SYW-6T=DSM 103992T=KCTC 52558T).

Legionella qingyii sp. nov., isolated from water samples in China.

Three Legionella-like strains, designed km488T, km489 and km521, were isolated from freshwater samples in China. Cells were Gram-stain-negative, rod-shaped and non-spore-forming. Growth was observed on BCYEα agar, but not on BCYEα agar without l-cysteine, chocolate agar with PolyViteX or Columbia blood agar. The major fatty acids (>5 %) of strains km488T, km489 and km521 were C16 : 0, anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The mip gene sequences (574 nt) showed the isolates were almost identical with more than 99.7 % sequence similarities, and closely matched to L. gormanii ATCC 33297T with 95.4-95.6 % sequence similarities. Phylogenetic analyses based on concatenated gene (16S rRNA, mip, rpoB and rnpB) sequences indicated that the isolates formed a distinct cluster along with L. gormanii within the genus Legionella. Matrix-assisted laser desorption ionization time-of-flight analyses also demonstrated a clear separation between the isolates and other closely and distantly related Legionella species. DNA-DNA hybridization studies demonstrated that the isolates were closely related (92.0 -95.0 % DNA-DNA relatedness) but differentiated from their phylogenetic neighbours (<70 % DNA-DNA relatedness). The whole genome of km488T was sequenced, and showed a G+C content of 37.8 mol%. Based on the findings from this polyphasic taxonomic study, the isolates are considered to represent a single novel species, for which the name Legionella qingyii sp. nov. is proposed. The type strain is km488T (KCTC 15636T=CCTCC AB 2018025T=NRBC 113223T).

Pseudofrancisella aestuarii gen. nov., sp. nov., a novel member of the family Francisellaceae isolated from estuarine seawater.

A Gram-negative, aerobic, non-motile and non-spore forming bacterium, designated strain SYSU WZ-2T, was isolated from an estuarine seawater sample. Growth of strain SYSU WZ-2T was observed at temperature range of 10-40° C (optimum, 32 °C), pH range of 6-10 (optimum, pH 7-8) and in the presence of up to 5.0% NaCl (w/v). The DNA G+C content of the novel strain was determined to be 30.1% (genome). The major polar lipids were found to be diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, an unidentified aminolipid, two unidentified aminophospholipids and two unidentified phospholipids. The major fatty acids were C18:0 3-OH (27.5%), C18:1ω9c (19.3%), C16:0 (17.0%) and C14:0 (12.9%). The respiratory quinone was found to be ubiquinone Q8. Pairwise comparison of the 16S rRNA gene sequence showed that strain SYSU WZ-2T shares high identities with members of the genera Francisella (94.8-95.9%) and Allofrancisella (93.8-94.2%). The phylogenetic dendrograms based on 16S rRNA gene sequences with the members of the family Francisellaceae showed that the strain SYSU WZ-2T formed a distinct phylogenetic lineage well separated from the members of the genera Francisella and Allofrancisella. MALDI-TOF mass spectrometric analysis also depicted a different profile for strain SYSU WZ-2T compared with those of members of the genera Francisella and Allofrancisella. Based on the above results and differences in phenotypic and chemotaxonomic features, strain SYSU WZ-2T is characterized to represent a new species of a novel genus, for which the name Pseudofrancisella aestuarii gen. nov., sp. nov. is proposed (type strain SYSU WZ-2T = KCTC 52557T = CGMCC 1.13718T).

Serratia microhaemolytica sp. nov., isolated from an artificial lake in Southern China.

A Gram-stain negative, facultatively anaerobic, rod-shaped, non-motile and non-spore forming bacterium, designated ZS-11T, was isolated from an artificial freshwater lake in Guangzhou city, Guangdong province, China. Growth of strain ZS-11T was observed at the temperature 18-42 °C (optimum 32-37 °C), pH 6.0-8.0 (optimum 7.0) and 0.5-3.0% (w/v) NaCl (optimum 0.5%, w/v), and also found to be enhanced in the presence of CO2. Pairwise comparison of 16S rRNA gene sequences showed that the strain shared high similarities with Serratia entomophila DSM 12358T (96.1%), Serratia ficaria DSM 4569T (96.0%), Serratia plymuthica DSM 4540T (96.0%), Rahnella victoriana FRB 225T (95.9%) and Rouxiella badensis DSM 100043T (95.8%). The phylogenomic dendrograms showed that strain ZS-11T formed a distinct cluster within the clade of the genus Serratia. The major fatty acids (> 20%) present in the cells were C16:0, C16:1ω7c/C16:1ω6c and C18:1ω7c/C18:1ω6c, which were consistent with those of S. entomophila CCUG 55496T and Serratia liquefaciens CCUG 9285T. The DNA G + C content for the genome was 49.3%. Based on these phenotypic and genotypic data, strain ZS-11T is considered to represent a new species of the genus Serratia, for which the name Serratia microhaemolytica sp. nov. is proposed. The type strain is ZS-11T (= CCTCC AB 2018040T = KCTC 62413T).