[Etiological characterization of invasive non-typhoid Salmonella strains in Guangdong Province from 2018 to 2022].

Objective: To understand the serotype distribution, drug resistance and molecular characterization of invasive non-typhoid Salmonella (iNTS) in Guangdong Province from 2018 to 2022 and provide scientific evidence for the prevention and treatment of blood flow infection caused by Salmonella. Methods: Serological identification, antimicrobial susceptibility testing, multilocus sequence typing (MLST), and whole genome sequencing were performed on Salmonella isolated from blood and stool samples in Guangdong from 2018 to 2022. Simultaneously, annotated the sequencing results for drug resistance genes and virulence factors by a microbial gene annotation system. Results: The 136 iNTS strains were divided into 25 serotypes, and Salmonella enteritidis accounted for 38.24% (52/136). The OR of other iNTS serotypes were calculated with Salmonella typhimurium as the control. The OR values of Oreninburg, Rysson, and Pomona serotypes were the highest, which were 423.50, 352.92, and 211.75, respectively. The drug resistance rate of iNTS was 0.74%-66.91%, which was lower than that of non-iNTS (3.90%-77.21%). The main iNTS of drug resistance were ampicillin and tetracycline, with resistance rates of 66.91% (91/136) and 50.00% (68/136), respectively, while the resistance rates to ciprofloxacin (5.88%,8/136), ceftazidime (5.88%,8/136), gentamicin (5.13%,7/136) and cefoxitin (0.74%, 1/136) were relatively low. iNTS carried a variety of drug-resistance genes and virulence factors, but no standard virulence factor distribution has been found. MLST cluster analysis showed that iNTS was divided into 26 sequence types, and ST11 accounted for 38.24% (52/136). Conclusions: The iNTS strains in Guangdong were dominated by Salmonella enteritidis, of which three serotypes, Oreninburg, Rison, and Pomona, may be associated with a higher risk of invasive infection during 2018 to 2022. iNTS was sensitive to clinical first-line therapeutic drugs (cephalosporins and fluoroquinolones), with highly diverse sequences and clear phylogenetic branches. ST11 was the local dominant clone group.

First co-infection case of melioidosis and Japanese encephalitis in China.

BACKGROUND: Melioidosis is endemic in Southeast Asia and northern Australia. Infection usually follows percutaneous inoculation or inhalation or ingestion of the causative bacterium, Burkholderia pseudomallei, which is present in soil and surface water in endemic regions. Japanese encephalitis (JE) is a vector-borne viral zoonosis caused by Japanese encephalitis virus (JEV), leading to epidemic encephalitis in Southeast Asia. Both B. pseudomallei and JEV have spread dominantly in the Hainan and Guangdong provinces in China. Here we reported the first case of co-infection of B. pseudomallei and JEV, which was discovered in Huizhou in the Guangdong province in June 2016. CASE PRESENTATION: A 52-year-old man was admitted to the hospital with acute febrile illness and headache, diagnosed as respiratory infection, central nervous system (CNS) infection, septicemia, and hepatic dysfunction. Based on B. pseudomallei-positive blood and cerebrospinal fluid (CSF) cultures, the patient was diagnosed with melioidosis and treated aggressively with antibiotics. However, the patient failed to make a full recovery. Further laboratory tests focused on CNS infection were conducted. The co-infection of B. pseudomallei and JEV was confirmed after the positive IgM antibodies of JEV were detected in both CSF and blood. After diagnosis of co-infection with B. pseudomallei and JEV, the patient was provided supportive care in hospital and recovered after approximately 3 weeks. CONCLUSION: Given the possibility of co-infection of B. pseudomallei and JEV, as well as variable case presentations, it is critical to enhance the awareness, detection, and treatment of co-infection in regard to melioidosis.

[Circulation and etiological characterization of Salmonella enterica serotype in human in Guangdong province, 2007-2016].

Objective: To understand the circulation, drug resistance and molecular characteristics of Salmonella1, 4, [5], 12: i:- in human in Guangdong province. Methods:Salmonella1, 4, [5], 12: i:- isolated from diarrhea patients in Guangdong during 2007-2016 were detected for drug resistance, genes and PFGE characteristics. Results: A total of 2 960 strains Salmonella1, 4, [5], 12: i: - were isolated from human diarrhea cases during this period. The positive rates of the isolation increased year by year. The male to female ratio of the infection cases was 1.58∶1, and the infection mainly occurred in infants and young children. Except imipenem, Salmonella1, 4, [5], 12: i: - was resistant to other 17 antibiotics to some extent. The drug resistant rates to ceftazidime, cefotaxime and ciprofloxacin increased from 2011 to 2016. Multi-drug resistance was serious, for example, the multi-drug resistant strains with ASSuT accounted for 70.62% (435/616) and the multi-drug resistant strains with ACSuGSTTm accounted for 27.11% (167/616). The lack of fljA, fljB and hin genes, as well as the retaining of iroB, STM2740, STM2757 genes, resulted in the unable expression of FljBenx gene with 8 different defection profiles. There were 934 different PFGE patterns observed in 2 347 strains, which displayed a relatively large fingerprint polymorphism. The major PFGE pattern was JPXX01. GD0226, which was found in 97 strains, accounting for 4.13% (97/2 347). The PFGE patterns in 168 Salmonella1, 4, [5], 12: i: - strains were consistent with that of Salmonella typhimurium. Conclusions:Salmonella1,4,[5], 12: i: - strains has become the major serotype of Salmonella that cause diarrhea in human in Guangdong. The multi-drug resistance of Salmonella1,4, [5], 12: i: - was serious, and since the defection of fljA, fljB and hin genes, the expression of FljBenx protein failed. The PFGE results were diverse, which displayed polymorphism in inheritance.

[Etiologic characteristics of Shigella sonnei strains isolated from some areas of Guangdong province and Guangxi Zhuang Autonomous Region of China, 2014-2016].

Objective: To investigated the etiologic characteristics of Shigella (S.) sonnei strains causing outbreaks and sporadic cases in some areas of Guangdong province and Guangxi Zhuang Autonomous Region during 2014-2016. Methods: Fourteen S. sonnei strains isolated from outbreaks and 6 S. sonnei strains from sporadic cases from Guangdong and Liuzhou of Guangxi Zhuang Autonomous Region were tested for antimicrobial resistance and analyzed by pulsed-field gel electrophoresis (PFGE). Six typical strains were selected for whole genome sequencing typing and compared with 51 strains isolated both at home and abroad from NCBI genome database. Results: The antibiotic resistance test indicated the isolates had high resistance rate to ampicillin, tetracycline, gentamicin, trimethoprim/sulfamethoxazole and nalidixic acid, while sensitive to azithromycin, chloromycetin and imipenem. PFGE showed high similarity (93.2%) among the strains isolated from different areas. The whole genome sequencing analysis also revealed that all the typical strains were clustered into a same evolution branch, close to some strains from Korea. Conclusions: The S. sonnei strains isolated from some areas of Guangdong and Guangxi Zhuang Autonomous Region showed high resistance to commonly used antibiotics, but they were sensitive to azithromycin, chloramphenicol and imipenem. The isolates in this study also showed similar PFGE patterns and close phylogenic evolution.

[Active etiological surveillance for foodborne diseases in Guangdong province, 2013-2014].

Objective: To study the infection status, serotypes, drug resistance and molecular characteristics of Salmonella, Shigella, Vibrio parahemolyticus, enterotoxigenic Escherichia (E.) coli (ETEC), pathogenic E. coli (EPEC), Shiga Toxin producing E. coli (STEC) and Enteroinvasive E. coli (EIEC) collected from diarrhea patients in Guangdong. Methods: The strains of Salmonella, Shigella, V. parahemolyticus and 4 kinds of E. coli isolated from foodborne diseases surveillance during 2013-2014 were collected to conduct serotyping, drug resistance test and pulsed-field gel electrophoresis (PFGE). Results: A total of 3 372 stains of pathogens were isolated from 57 834 stool samples during 2013-2014, the overall positive rate was 5.83% and the positive rate of Salmonella was highest, followed by that of V. parahemolyticus, 4 kinds of E. coli and Shigella. And 3 213 strains of Salmonella were divided into 143 serotypes. The most prevalent serotypes were Salmonella typhimurium, 4, 5, 12: i:-, Enteritidis, Stanley and Derby. Salmonella was sensitive to cephalosporin and fluoroquinolones, and showed significant differences in drug resistance rate among different serotypes. In top 10 common serotypes, S. enteritidis and S. derby were most resistant to cephalosporin and ciprofloxacin respectively. PFGE was performed for 2 289 strains of Salmonella, showing distribution diversity and significant fingerprint polymorphisms. The 85 strains of V. parahemolyticus were divided into 10 serotypes, O3∶K6 (61.18%) was the most common serotype, followed by O4∶K8. The results showed that the carrying rate of virulence genes tdh (81.18%) was high, while the carrying rate of trh was low (7.06%), and there were 10 strains carrying no the two kinds of virulence genes. The sensitive rate of V. parahemolyticus to imipenem, nalidixic acid, SMZ-TMP, chloramphenicol and tetracycline were more than 95%. Thirteen strains of Shigella were detected, including 9 strains of Shigella sonnei, 3 strains of Shigella flexneri and 1 strains of Shigella bogdii. The strains all showed sensitivity to ceftazidime, ciprofloxacin and chloramphenicol (76.92%). There were 86 strains of E. coli detected, including 29 strains of ETEC (33.72%), 27 strains of EPEC (31.39%), 27 strains of STEC (31.39%) and 3 strains of EIEC (3.48%). Conclusions: In the active etiological surveillance for foodborne diseases in Guangdong during 2013-2014, the detection rate of Salmonella was highest (5.57% ), followed by that of V. parahemolyticus, 4 kinds of E. coli and Shigella. Salmonella, V. parahemolyticus and Shigella were sensitive to cephalosporin and fluoroquinolones. Clustered cases of Salmonella infection were found in the surveillance, but no outbreaks occurred.

[Study on Salmonella serotyping by use of Microsphere-based Liquid Array method].

OBJECTIVE: To understand the effect of serotyping on Salmonella isolates, by use of Microsphere-based Liquid Array method, among diarrhea patients, in Guangdong. METHODS: Salmonella isolated from humans in Guangdong province were serotyped on the Microsphere-based Liquid Array platform with SSA kit. RESULTS: A total of 4 942 Salmonella strains with 189 serotypes, were identified in Guangdong province in 2010-2014. The top 100 serotypes accounted for 98.08% (4 847/4 942) of all the strains. 98% of the top 100 species serotypes could completely be serotyped with SSA kit. In order to detect O antigen among 198 isolates with SSA kit, 181 strains were carrying the O antigen, with the coincidence rate as 100%. However, under the SSA, 98.32% (528/537) of the H antigen could be detected and were consistent with the traditional serum agglutination test. The coincidence rate of fljB gene was 93.09% (175/188), with false negative rate and false positive rate of fljB gene as 7.35% (9/134) and 7.41% (4/54) respectively. The coincidence rate of sdf gene and Vi gene were 100%. 11 out of the 12 Salmonella strains could not be serotyped under the traditional methods but were successfully serotyped by the molecular serotyping method. CONCLUSIONS: Using the SSA kit, more than 96% of the anthropogenic Salmonella strains could be serotyped in Guangdong province. Comparing with the traditional methods, the coincidence rate of serotyping appeared over 98% . Under the Microsphere-based Liquid Array techniques, the molecular serotyping method appeared faster and more accurate on Salmonella serotyping than those traditional methods.

Genetic relatedness of selected clinical Vibrio cholerae O139 isolates from the southern coastal area of China over a 20-year period.

Vibrio cholerae O139 emerged as a causative agent of epidemic cholera in 1992 in India and Bangladesh, and was subsequently reported in China in 1993. The genetic relatedness and molecular characteristics of V. cholerae O139 in Guangdong Province, located in the southern coastal area of China, remains undetermined. In this study, we investigated 136 clinical V. cholerae O139 isolates from 1993 to 2013 in Guangdong. By conventional PCR, 123 (90·4%) isolates were positive for ctxB, ace and zot. Sequencing of the positive amplicons indicated 113 (91·7%) isolates possessed the El Tor allele of ctxB (genotype 3); seven carried the classical ctxB type (genotype 1) and three harboured a novel ctxB type (genotype 5). With respect to tcpA, 123 (90·4%) isolates were positive for the El Tor allele. In addition, pulsed-field gel electrophoresis (with NotI digestion) differentiated the isolates into clusters A and B. Cluster A contained seven of the non-toxigenic isolates from 1998 to 2000; another six non-toxigenic isolates (from 1998 and 2007) and all of the toxigenic isolates formed cluster B. Our results suggest that over a 20-year period, the predominant O139 clinical isolates have maintained a relatively tight clonal structure, although some genetic variance and shift has occurred. Our data highlight the persistence of toxigenic V. cholerae O139 in clinical settings in the southern coastal area of China.

Comparing the ability of luminex xMAP(®) salmonella serotyping assay and traditional serotyping method for serotyping salmonella isolated from southern Chinese population.

AIMS: To investigate whether the Luminex xMAP(®) Salmonella Serotyping Assay (xMAP SSA) is applicable to serotype Salmonella from humans in southern China. METHODS AND RESULTS: Two hundred and five Salmonella isolates from diarrhoea patients were serotyped by xMAP SSA in parallel with the traditional serotyping. Forty serotypes were identified among 205 isolates; the most prevalent serotypes identified were Salmonella Enteritidis, Salmonella Stanley, Salmonella I 4,5,12:i:-, and Salmonella Typhimurium. One hundred and ninety-five (95·1%, 195/205) isolates were serotyped completely by xMAP SSA, while 10 stereotypes were partially detected as they were not included in the assay. The xMAP SSA correctly identified 192 (98·5%, 192/195) isolates. Five nonmotile and three monophasic strains, which possessed flagellar antigen genes that weren't expressed, were completely serotyped by xMAP SSA; however, these isolates were left undetected by the traditional method. CONCLUSION: The xMAP SSA used in the study is a microsphere-based, molecular assay that could rapidly and accurately serotype Salmonella. It is suitable to identify the serovars of Salmonella in southern China. SIGNIFICANCE AND IMPACT OF THE STUDY: The xMAP SSA, with high-throughput characteristics, provides an accurate and rapid serotyping system that dramatically strengthens the capability of clinical and public health laboratories for Salmonella serotyping.