Objective To evaluate the feasibility of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for identification and cluster analysis of Streptococcus suis. Methods Eighteen clinical isolates biochemically identified as Streptococcus suis were pre-treated by smearing formic acid method and formic acid-acetonitrile extraction method. The identification results and protein profiles of MALDI-TOF-MS method were analyzed and compared. A self-constructed database of profiling with better pretreatment method was established, and cluster analysis was performed on the 18 strains of Streptococcus suis. At the same time, PFGE homology analysis was performed to compare the results of the two genotyping. Results Both pretreatment methods could accurately identify Streptococcus suis with scores above 2.1. The protein fingerprint of formic acid and acetonitrile extraction method had a smoother baseline, fewer miscellaneous peaks and more identifiable ion peaks. Comparison of the results of homology typing showed that the homology results of MALDI-TOF-MS were significantly different from those of PFGE. Conclusion MALDI-TOF-MS can accurately identify Streptococcus suis for the strains pre-treated with formic acid method or formic acid-acetonitrile extraction method, and the formic acid-acetonitrile extraction method can obtain a better protein mapping. MALDI-TOF-MS can play a certain role in typing, but it still has some limitations and cannot completely replace the PFGE.

Objective To study the biological characteristics of clinical isolates of human infected Streptococcus suis type 2 in Zhongshan City from 2016 to 2019 and classify the strains using pulse-field gel electrophoresis (PFGE), and to provide a scientific basis for clinical prevention and treatment of porcine streptococcus suis disease. Methods Twelve strains of Streptococcus suis type 2 were collected from 2016 to 2019 and identified by automatic bacterial identification instrument. The carrying status of five major virulence genes of Streptococcus suis was detected by nucleic acid and protein analyzer, including capsular polysaccharide (cps2J), lysozyme-releasing protein (mrp), hemolysin (sly), glutamate dehydrogenase (gdh), and extracellular factor (ef). The susceptibility of Streptococcus suis to 12 kinds of commonly used antibiotics was determined by the broth microdilution method, and the homology analysis was carried out by PFGE method. Results Twelve strains of Streptococcus suis type 2 were divided into four virulence genotypes, mainly mrp-/sly+/ef+/cps2J+/gdh+ (6strains) and mrp-/sly-/ef+/cps2J+/gdh+(4strains). Drug susceptibility test results showed that 12 strains of Streptococcus suis type 2 were resistant to erythromycin, tetracycline and clindamycin, and they all were multi-resistant strains. According to the classification results of PFGE, the 12 strains were classified into 7 PFGE types based on 100% similarity coefficient. The PFGE band types of Streptococcus suis in the same year had high homology. Conclusion The virulence genotypes of 12 clinical isolates of human infected type 2 Streptococcus suis in Zhongshan from 2016 to 2019 are diverse, and the strains are resistant to multiple antibiotics. Most strains in the same year are the same clone strains. PFGE genotypes are not correlated with virulence genotypes and drug resistance spectrum.

Objective To analyze the genetic characteristics of Legionella pneumophila isolated from cooling water of central air conditioning system in public places in Zhongshan from 2012 to 2018, and to understand the spatiotemporal distribution of homologous strains, in order to provide evidence for the prevention, control and traceability of Legionella pneumophila infection. Methods Eighty-five Legionella pneumophila strains were isolated for serotype identification, and the molecular typing of the 85 isolates was performed using pulsed field gel electrophoresis (PFGE). The strain location data was converted into latitude and longitude coordinates by GIS geocoding technology. The converted location data was overlaid on the map of Zhongshan City, mapping the molecular typing distribution of clusters using Qgis2.18.11 spatial processing software. Results Eighty-five strains of Legionella pneumophila included 9 serotypes, and the highest proportion was LP1, accounting for 61.18% (52/85). According to the similarity of 100%, 85 strains of Legionella pneumophila were divided into 56 patterns of PFGE bands (T1-T56), with 3 types being dominant. Same serotype of Legionella pneumophila strains showed diverse PFGE patterns. Different serotypes of Legionella pneumophila strains were basically identified as different PFGE patterns, while some were identified as same PFGE pattern. According to over 85% similarity, 8 clusters (A-H) were designated, strains of which were distributed in 12 districts. PFGE clustering clusters did not display obvious temporal and regional distribution differences, nor did they have temporal and regional clustering distributions. Conclusion Strains of Legionella pneumophila isolated from cooling water of central air conditioning system in public places in Zhongshan from 2012 to 2018 showed genetic diversity, and the main serotype was LP1. Isolates of clusters did not exist in different years or regions.

Objective To investigate the etiological characteristics of food poisoning isolates of Vibrio parahaemolyticus (VP) from 2019 to 2021 in Zhongshan City. Methods A total of 37 strains of Vibrio parahaemolyticus isolated from 8 food poisoning incidents in Zhongshan City from 2019 to 2021 were collected, including 1 residual food isolate and 36 human isolates. The genetic correlation of Vibrio parahaemolyticus food poisoning isolates in this region was analyzed by serological typing, virulence gene detection (TLH, TDH, and TRH), drug sensitivity test, pulsed field gel electrophoresis (PFGE) and multipoint sequence typing (MLST). Results The 37 strains of Vibrio parahaemolyticus were divided into 4 serotypes: O3:K6, O10:K4, O4:K8, and O4:KUT. The tdh+ and trh- were the main virulence genotypes, accounting for 97.30% (36/37). The drug resistance rate of cefazolin was 40.54% (15 strains R, 22 strains I), and no multidrug-resistant strains were found. The 37 VP strains were divided into 23 PFGE types and 6 cluster groups, with correlation coefficients ranging from 60.4%-100%. The multipoint sequencing typing showed that the 37 VP strains were divided into 9 ST types and 3 complex groups, of which ST3 type was the main type (23 strains, 62.1%). Conclusion This study has found that the dominant virulence types of Vibrio parahaemolyticus food poisoning isolates in Zhongshan City from 2019 to 2021 are tdh⁺ and trh^-, and 37 representative strains can be divided into 6 PFGE clusters and 9 ST types with MLST type being mainly ST3. This study has identified the rare serotype O10:K4 which has caused an increase in the proportion of food poisoning events, suggesting that we should strengthen detection and be alert to the risk of continued local epidemics of new rare serotype strains.