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).

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).

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).

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).

Fastidiosibacter lacustris gen. nov., sp. nov., isolated from a lake water sample, and proposal of Fastidiosibacteraceae fam. nov. within the order Thiotrichales.

AFrancisella-like bacterium, designated strain SYSU HZH-2T, was isolated from a water sample collected from Haizhu Lake, Guangzhou, China. The bacterium was fastidious, and required an exogenous source of l-cysteine for its growth on artificial media. Cells were Gram-stain-negative, coccobacilli, non-motile and non-spore-forming. The strain shared highest 16S rRNA gene sequence similarities with Cysteiniphilum litorale SYSU D3-2T (94.6 % identity), Fangia hongkongensis UST040201-002T (93.2 %) and Caedibacter taeniospiralis 51T (91.6 %). This strain possessed ubiquinone-8 as the respiratory quinone; diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol as the known polar lipids, and anteiso-C15 : 0 and C18 : 0 as the major fatty acids (>10 % of total fatty acids). The dendrograms based on 16S rRNA gene sequence analysis showed that it formed a separate cluster along with Cysteiniphilum litorale SYSU D3-2T, Caedibactertaeiniospiralis 51T and Fangia hongkongensis UST040201-002T. Based on the 16S rRNA gene sequence identity and differences in other phenotypic characteristics, the strain is considered to represent a novel species of a novel genus, for which the name Fastidiosibacter lacustris gen. nov., sp. nov. is proposed. The type strain of the type species Fastidiosibacter lacustris is SYSU HZH-2T (=NBRC 112274T = CGMCC 1.15950T). Additionally, the new taxon along with the genera Caedibacter, Cysteiniphilum and Fangia (family unassigned) were distinctly separated from the related families Francisellaceae, Piscirickettsiaceae and Thiotrichaeae in the phylogenetic trees. Therefore, we proposed a new family Fastidiosibacteraceae fam. nov. within the order Thiotrichales to accommodate these four genera.

Timing of orthognathic surgery on the changes of oral health-related quality of life in Chinese orthognathic surgery patients.

INTRODUCTION: Although the association between orthognathic surgery and oral health-related quality of life (OHRQoL) has been explored, few studies have been carried out to describe the influence of surgery type on perceived OHRQoL. The aim of this study was to evaluate the difference of OHRQoL between a surgery-first approach and an orthodontics-first approach, using the 14-item Oral Health Impact Profile questionnaire (Chinese version). METHODS: Fifty adult Chinese orthodontic patients who received either surgery-first or orthodontics-first treatment completed 5 distinct sections of the questionnaire. Chi-square tests were used to compare categorical variables. All analyses were carried out with Stata software (version 11.2; StataCorp, College Station, Tex). RESULTS: The quality of life significantly improved after treatment in both groups. However, overall scores were highest before treatment in the surgery-first group but increased significantly from before treatment to 6 months after surgery and then significantly decreased after surgery in the orthodontics-first group. There were relatively lower scores in the surgery-first group than in the orthodontics-first group before surgery, at 12 months after beginning orthodontics, and at the end of treatment, although these differences did not reach significant levels. CONCLUSIONS: Both treatment methods can obtain the same results. The timing of the orthognathic approach did not affect the final OHRQoL in Chinese orthognathic surgery patients.

Self-ligating brackets and their impact on oral health-related quality of life in Chinese adolescence patients: a longitudinal prospective study.

INTRODUCTION: Although the associations between orthodontic and oral health-related quality of life (OHRQOL) have been explored, little research has been done to address the influence of brackets type on perceived OHRQOL. The aim of this study was to assess whether the levels of OHRQOL in Chinese adolescence patients were influenced by the type of brackets. MATERIALS AND METHODS: One hundred fifty Chinese orthodontic adolescence patients completed the 14-item Oral Health Impact Profile (OHIP-14, Chinese version) at five distinct intervals: after insertion of the fixed appliance at 1 week (T1), 1 month (T2), 3 months (T3), and 6 months (T4); and after treatment (T5). RESULTS: Patients with self-ligating brackets were associated with less pain and discomfort at any intervals compared with conventional brackets, but no significant difference of overall OHIP-14 scores could be found between two groups. Moreover, in both groups, overall scores at T1 and T2 were significantly higher than the scores at any other intervals in both groups. CONCLUSIONS: The type of orthodontic appliance did not affect oral health-related quality of life in Chinese adolescence patients.

Comparative Genomic Analysis Reveals Genetic Diversity and Pathogenic Potential of Haemophilus seminalis and Emended Description of Haemophilus seminalis.

Haemophilus seminalis is a newly proposed species that is phylogenetically related to Haemophilus haemolyticus. The distribution of H. seminalis in the human population, its genomic diversity, and its pathogenic potential are still unclear. This study reports the finding of our comparative genomic analyses of four newly isolated Haemophilus strains (SZY H8, SZY H35, SZY H36, and SZY H68) from human sputum specimens (Guangzhou, China) along with the publicly available genomes of other phylogenetically related Haemophilus species. Based on pairwise comparisons of the 16S rRNA gene sequences, the four isolates showed <98.65% sequence identity to the type strains of all known Haemophilus species but were identified as belonging to H. seminalis, based on comparable phenotypic and genotypic features. Additionally, the four isolates showed high genome-genome relatedness indices (>95% ANI values) with 17 strains that were previously identified as either "Haemophilus intermedius" or hemin (X-factor)-independent H. haemolyticus and therefore required a more detailed classification study. Phylogenetically, these isolates, along with the two previously described H. seminalis isolates (a total of 23 isolates), shared a highly homologous lineage that is distinct from the clades of the main H. haemolyticus and Haemophilus influenzae strains. These isolates present an open pangenome with multiple virulence genes. Notably, all 23 isolates have a functional heme biosynthesis pathway that is similar to that of Haemophilus parainfluenzae. The phenotype of hemin (X-factor) independence and the analysis of the ispD, pepG, and moeA genes can be used to distinguish these isolates from H. haemolyticus and H. influenzae. Based on the above findings, we propose a reclassification for all "H. intermedius" and two H. haemolyticus isolates belonging to H. seminalis with an emended description of H. seminalis. This study provides a more accurate identification of Haemophilus isolates for use in the clinical laboratory and a better understanding of the clinical significance and genetic diversity in human environments. IMPORTANCE As a versatile opportunistic pathogen, the accurate identification of Haemophilus species is a challenge in clinical practice. In this study, we characterized the phenotypic and genotypic features of four H. seminalis strains that were isolated from human sputum specimens and propose the "H. intermedius" and hemin (X-factor)-independent H. haemolyticus isolates as belonging to H. seminalis. The prediction of virulence-related genes indicates that H. seminalis isolates carry several virulence genes that are likely to play an important role in its pathogenicity. In addition, we depict that the genes ispD, pepG, and moeA can be used as biomarkers for distinguishing H. seminalis from H. haemolyticus and H. influenzae. Our findings provide some insights into the identification, epidemiology, genetic diversity, pathogenic potential, and antimicrobial resistance of the newly proposed H. seminalis.

A feasible method to improve adherence of Hawley retainer in adolescent orthodontic patients: a randomized controlled trial.

BACKGROUND: Retention is an important component of orthodontic treatment; however, poor compliance with retainer use is often encountered, especially in adolescents. The purpose of this study was to prove the hypothesis that verbal instructions combined with images showing the severe consequences of poor compliance can increase retainer use. METHODS: This study was a randomized controlled trial. The sample was recruited from Wenzhou, People's Republic of China, between February 2013 and May 2014, and 326 participants were randomized into three groups. Patients and parents in Group A (n=106) were given routine retainer wear instructions only; in Group B (n=111), images illustrating the severe consequences of poor compliance with Hawley retainer use were shown to patients, combined with routine instructions; and in Group C (n=109), images illustrating the severe consequences of poor compliance with Hawley retainer use were shown to patients and parents, combined with routine instructions. Three months after debonding, questionnaires were used to investigate daily wear time and the reasons for poor compliance. Differences in means between the groups were tested by one-way analysis of variance. RESULTS: The mean daily wear time in Group C (15.09±4.13 hours) was significantly greater than in Group A (12.37±4.58 hours, P<0.01) or Group B (13.50±4.22 hours, P<0.05); the mean daily wear time in Group B was greater than in Group A, but was not significant (P=0.67). Reasons for nonusage were forgetting to wear the retainer (51%) and finding the retainer bothersome to frequently insert and remove (42%). CONCLUSION: Verbal instructions combined with images showing the severe consequences of poor compliance can increase retainer use. Parents play an important role in compliance with retainer use in adolescent patients.

Use of Q methodology to assess the concerns of adult female individuals seeking orthodontic treatment.

BACKGROUND: Orthodontic treatment may cause functional restrictions, discomfort, and pain, which may lead to dental anxiety and noncooperation among patients. This study aimed to assess the concerns of adult female patients with respect to such treatment. PATIENTS AND METHODS: We conducted an explorative study using Q methodology among 40 adult female patients with different educational and social backgrounds in Wenzhou, People's Republic of China. We asked participants to rank a set of 41 statements about seeking orthodontic treatment on an 11-point scale from "agree most" to "disagree most". The collected data were analyzed using the PQ Method 2.35 program. We extracted significant viewpoints using centroid factor extraction and varimax rotation. RESULTS: We identified major factors based on how the patients ranked statements. Patients in group 1 worried about lack of information about orthodontic treatment, and may have suffered from dental phobia; patients in group 2 were all single women, and they were worried that the braces might lower their chances of finding a partner; patients in group 3 worried about appearance and speech with braces; and patients in group 4 worried about cost, pain, and dental hygiene. The remaining participants who had other viewpoints did not load to any of these four groups. CONCLUSION: The concerns of adult female individuals seeking orthodontic treatment are complex. A significant feature of this study was using Q methodology to analyze the psychological characteristics of the patients. This study identified four typical characterizations that are associated with each group, and our findings may aid orthodontists in improving doctor-patient relationships.

[Soft tissue changes of upper lip in patients with class I malocclusion with different premolar extraction patterns].

OBJECTIVE: The purpose of this retrospective cephalometric study was to compare changes of the upper lip soft tissue in patients with Class I malocclusion after extraction of either first premolars or second premolars. METHODS: The pretreatment and posttreatment records of 50 patients were used in the study. And 25 patients involved extraction of first premolars and 25 patients involved the extraction of second premolars. The lateral cephalometric radiographs taken before and after treatment were traced and measured. 15 typical values of soft tissue were measured in Winceph 7.0 software and statistical analysis was carried by SPSS 10.0 software. RESULTS: 1) The horizontal and vertical lines were not significant different in the two extraction group before the treatment. But after the treatment the vertical changes of upper lip (A'-X, UL-X) were more apparent in the second premolar extraction group (P < 0.05). 2)In the soft tissue measurements, the values of Sn-A'-UL in the first premolar extraction group were less than those in the second premolar extraction group significantly before the treatment. After the treatment all the patients had a significant increase in length of upper lip. At the same time, UL-EP, A'-EP distances were diminished obviously (P < 0.05). But the changes were not significant difference between the two premolar extraction groups (P > 0.05). CONCLUSION: The upper lips both changed remarkably after the treatment and there was no significant difference between the two premolar extraction groups.

[Soft tissue changes of chin with different premolar extraction patterns].

PURPOSE: The purpose of this retrospective cephalometric study was to compare the change of chin soft tissue in patients with Class I malocclusion after extraction of either first premolars or second premolars. METHODS: The pretreatment and posttreatment records of 50 patients were used in the study. Twenty-five patients underwent extraction of first premolars and 25 patients underwent extraction of second premolars. All the patients were skeletal Class I and matched by the gender, age, facial divergence and crowding. The lateral cephalometric radiographs taken before and after treatment were traced and measured. The data was analyzed using SPSS 10.0 software package for Student's t test. RESULTS: (1)The horizontal and vertical changes was both significant in the second extraction group compared with first extraction group, and the horizontal change was more apparent. But only vertical change was apparent in the first extraction group. (2)In the soft tissue measurements, the value of soft tissue facial angle and B'-Pos-FH in the first extraction group was less than those in the second extraction group significantly. After treatment all the patients had an increase in S-Ns-Pos angle, soft tissue facial angle, B'-Pos-FH and depth of mentolabial groove. At the same time, soft tissue convexity angle was diminished obviously. But the change was not significant between the two premolar extraction groups. CONCLUSIONS: The growth pattern tends to vertical pattern in the first premolar extraction group and the horizontal growth of chin is more obvious in the second premolar extraction group. There is no significant difference of chin soft tissue changes between the two premolar extraction groups.

[Study of the crestal bone height and root length technique change of impacted canines treated by modified Nance arch technique].

PURPOSE: The purpose of this study was to evaluate the crestal bone height and root length change after orthodontic treatment of impacted canines with modified Nance arch technique. METHODS: 8 maxillary impacted canines were treated with modified Nance arch technique. The canines of other side which were naturally erupted were used for self-control. Pre-treatment and post-treatment panoramic radiographs were measured and the following data was collected: (1)The distance of the impacted canine migration and the angle change of the impacted canine; (2)Crestal bone height and root length between previously impacted canines and adjacent lateral incisors and premolars(experimental)and contralateral control teeth. The crestal bone height and root length were compared using a paired t test with SPSS11.0 software package. RESULTS: The mean distance of the impacted canine migration was 18.43 mm and the mean angle change on the panoramic radiograph was 32.7 degrees. No difference was found in the crestal bone height and the root length between the adjacent lateral incisors and premolars(experimental) and the contralateral control teeth(P>0.05). CONCLUSION: The combination of modified Nance arch with straight wire arch technique is a valid method to treat maxillary impacted canines.