Pathogenic detection and molecular tracing of a bacillary dysentery outbreak by Shigella sonnei in Huainan city / 中华预防医学杂志

Objective: To understand the virulence gene and drug resistance profile of Shigella sonnei outbreak in Huainan city, and conduct pathogenic traceability analysis. Methods: Water samples and feces related to an infectious diarrhea outbreak in Huainan city in August 2020 were collected for multiple pathogen detection. Virulence gene, drug sensitivity, pulse-field gel electrophoresis and whole genome sequencing of Shigella isolates were analyzed respectively. Results: 38 strains of Shigella sonnei were detected in 56 samples of mucilage feces with a positive rate 67.86%, and all serotypes were Shigella sonnei Phase I. Three strains of Shigella sonnei were detected by fluorescence PCR in the Gram-negative (GN) bacterial enrichment solution of terminal water and well water. Virulence genes were ipaH positive (38), ipaH/ial (31) and ipaH/ial/sen positive (1), respectively. The drug resistance spectrum showed that 9 of 14 antibiotics were 100% resistant, and only imipenem, chloramphenicol, ceftazidime and ciprofloxacin were effective drugs. XbaⅠ restriction enzyme map type of 36 isolates was completely consistent, and the ST type analysis of 3 strains was ST152. Whole genome sequencing and analysis verified that the outbreak was caused by a single clonal group of strains, and revealed that the isolates of the outbreak were clustered into a large cluster with 3 Chinese strains and 1 Korean strain in the database, far away from the strains of other countries. Conclusion: The outbreak is caused by a single clone of Shigella sonnei, which are low virulence strains and have multiple drug resistance.

Caracterización molecular de aislamientos de Shigella sonnei recuperados en el programa de vigilancia por el laboratorio de la enfermedad diarreica aguda en Colombia / Molecular characterization of Shigella sonnei isolates recovered by the Laboratory Surveillance Program for Acute Diarrheal Disease in Colombia

Introducción. En Colombia, Shigella sonnei es uno de los serotipos más frecuentemente aislados (53,4 %) de muestras clínicas humanas asociadas a la enfermedad diarreica aguda. La identificación de patrones de restricción del ADN mediante electroforesis en gel de campo pulsado constituye la base de la vigilancia molecular de S. sonnei . Objetivo. Establecer la base de la vigilancia molecular de S. sonnei en Colombia mediante electroforesis en gel de campo pulsado. Materiales y métodos. Se estudiaron 102 de los 2.048 aislamientos de S. sonnei remitidos por la Red Nacional de Laboratorios entre 1997 y marzo del 2013; la selección se hizo de acuerdo con el patrón de resistencia antimicrobiana, el origen de la muestra y la relación con brotes. Se determinó el patrón genético mediante electroforesis en gel de campo pulsado con las enzimas de restricción XbaI y Blnl, según el protocolo de la red PulseNet International. El análisis de los patrones electroforéticos se hizo con el programa GelCompar II, versión 4.0. Resultados. Se obtuvieron 42 patrones electroforéticos con una similitud de 70 a 100 %. El patrón más frecuente fue COIN08J16X01.0017 (17,6 %), seguido por los patrones COIN04J16X01.0004 (9,8 %) y COIN02J16X01.0002 (5,8 %), y el 66,8 % restante se asoció con otros patrones electroforéticos. El análisis de brotes demostró la relación genética de cada brote con 100 % de similitud en la identificación; el patrón más frecuente en los brotes fue el COIN08J16X01.0017 (17,1 %). Conclusión. Se estableció la base de datos genotípicos de aislamientos de S. sonnei a nivel nacional mediante electroforesis en gel de campo pulsado; se incluyeron los 42 patrones únicos identificados en este estudio.

Introduction: In Colombia, Shigella sonnei is one of the most frequently isolated serotypes (53.4%) in human clinical samples associated with diarrheal acute disease. The identification of DNA restriction patterns by pulsed field gel electrophoresis is the basis for the molecular surveillance of S. sonnei . Objective: To establish the basis for the molecular surveillance of S. sonnei in Colombia using pulsed-field gel electrophoresis. Materials and methods: We studied 102 of 2,048 S. sonnei isolates referred by the National Laboratory Network between 1997 and March, 2013; the selection was made according to the antimicrobial multiresistance profile, the source of samples, and the relation to outbreaks. The genetic profile was determined by pulsed field gel electrophoresis using the restriction enzymes XbaI and BlnI in accordance with the PulseNet International protocol. The electrophoretic patterns were analyzed with the GelCompare II, version 4.0 software. Results: We obtained 42 electrophoretic patterns with a 70% to 100% similarity. The most frequent pattern was COIN08J16X01.0017 with 17.6%, followed by patterns COIN04J16X01.0004 with 9.8%, and COIN02J16X01.0002 with 5.8%, while the remaining 66.8% was associated with other electrophoretic patterns. The analysis of 10 outbreaks demonstrated their genetic relation with a 100% of similarity; the most frequent pattern in outbreaks was COIN08J16X01.0017 with 17.1%. Conclusion: The genotypic database for Shigella sonnei isolates was established using pulsed field gel electrophoresis including the 42 unique patterns identified in this study.

Drug resistance profiles and clonality of sporadic Shigella sonnei isolates in Ankara, Turkey

The aims of this study were to investigate drug resistance rates, types of extended spectrum beta lactamases (ESBLs), and molecular epidemiological characteristics of 43 Shigella sonnei isolates. Ampicillin-sulbactam, amoxicillin-clavulanate, chloramphenicol, and ciprofloxacin were the most active antibiotics. Five isolates harbored blaSHV-12, blaTEM-1 and blaCTX-M-15. More than 90% of the isolates had an indistinguishable pulsotype.

Colicin type 7 produced by majority of Shigella sonnei isolated from Thai patients with diarrhoea

Thirty one out of 153 strains of Shigella sonnei isolated from Thai patients with diarrhoea showed antibacterial activity against S. sonnei by agar well diffusion method. All of them harbor plasmids with the genetic determination of colicin type 7 (Js) gene but without colicin E and colicin U gene. The PCR product obtained from strain 35/44 was shown to be the gene for colicin type 7 lytic protein (cja). The partially purified bacteriocin (PPB) containing colicin type 7 of strain 35/44 was prepared and used for characterization. The antibacterial activity of PPB against a total of 17 selected Gram-positive and Gram-negative bacteria was tested. It was found that PPB of strain 35/44 was active against E. coli O157, S. sonnei and S. boydii. The sensitivity of PPB from this strain to proteinase K, trypsin and α-chymotrypsin suggests the proteinaceous nature of these antimicrobial substances. Therefore, this isolated bacterium can be regarded as bacteriocin producing bacteria. The bacteriocin produced by this isolated S. sonnei was heat stable as evidenced by its ability to maintain the activity at 80 °C for 60 min. In addition, it was stable within a wide range of pH (3-9). The molecular weight of colicin type 7 from isolated S. sonnei strain 35/44 analyzed by SDS-PAGE was 54.4 kDa composing of at least five subunits. It is to our knowledge; the first report of Thai patients with diarrhoea that S. sonnei isolated from them contained colicin type 7.

Occurrence of class 2 integrons among multi-drug resistant Shigella sonnei isolated from Tehran, Iran in 2005

Shigellosis is one of the major causes of morbidity in children with diarrhea in Iran. Integrons play an important role in the evolution and dissemination of multidrug resistance in gram-negative bacteria. The occurrence of integrons among Shigella spp. is frequently reported throughout the world. The aim of this study was to assess the occurrence of class 2 integrons among the multi drug resistant S. sonnei isolated from Iranian children in 2005. The study was conducted in two major pediatric hospitals in Tehran, Iran. Fecal specimens and rectal swab collected from patients were cultured and identified as Shigella by the conventional methods. Antimicrobial susceptibility test was performed according to the standard CLSI guideline. Multi-drug resistant isolates of S. sonnei were further examined for the presence of class 2 integron by PCR using specific primers. Amplicons were confirmed by restriction endonuclease analysis. A total of 83 multi-drug resistant S. sonnei strains were isolated. Of these, 45 [54%] exhibited a class 2 integron of 2.1 kbp, and 34 [41%] a class 2 integron of 1.3 kbp. Class 2 integrons were not detected in four isolates. The results showed an increased occurrence of class 2 integron carrying S. sonnei isolated from children in Tehran in 2005

Fatality due to shigellosis with special reference to molecular analysis vi Shigella sonnet strains isolated from the fatal cases

Shigellosis as a global human health problem is more severe than other forms of gastroenteritis and causes over a million deaths in developing countries worldwide annually. Fatality due to shigellosis is usually due to dehydration and two-third of fatalities are seen among children. The aim of current study was to describe fatal cases of shigellosis due to infection with Shigella sonnei and S.flexneri. We investigated the fatal cases of shigellosis among all children with acute diarrhea admitted to Children's Medical Center, Tehran, Iran. Bacterial isolation and identification was achieved according to standard bacteriological methods. Antibiotic susceptibility tests, plasmid profiling and ribotyping were performed to investigate the clonal relationship among the isolates. Among 1200 children with acute diarrhea, 140 [12.7%] cases had shigellosis. Of these, three patients died. No signs of severe dehydration were observed among the fatal cases. The symptoms were not improved following antibiotic therapy and all three cases died after 24 h of hospitalization despite receiving intensive treatments. Stool cultures yielded S.flexneri and S. sonnei for one and two cases, respectively. The isolates were resistant to streptomycin, ampicillin, and sulfamethoxazole-trimethoprim. 5. sonnei strains were further studied and showed a single pattern of antibiotic susceptibility and ribotyping. Mortality due to species other than 5. dysenteriae is rare, however, in current study we found S. sonnei and S.flexneri as the cause of fatality among pediatric patients during the study

Epidemiological characterization of resistance and PCR typing of Shigella flexneri and Shigella sonnei strains isolated from bacillary dysentery cases in Southeast Brazil

Shigella spp are Gram-negative, anaerobic facultative, non-motile, and non-sporulated bacilli of the Enterobacteriaceae family responsible for "Shigellosis" or bacillary dysentery, an important cause of worldwide morbidity and mortality. However, despite this, there are very few epidemiological studies about this bacterium in Brazil. We studied the antibiotic resistance profiles and the clonal structure of 60 Shigella strains (30 S. flexneri and 30 S. sonnei) isolated from shigellosis cases in different cities within the metropolitan area of Campinas, State of São Paulo, Brazil. We used the following well-characterized molecular techniques: enterobacterial repetitive intergenic consensus, repetitive extragenic palindromic, and double-repetitive element-polymerase chain reaction to characterize the bacteria. Also, the antibiotic resistance of the strains was determined by the diffusion disk method. Many strains of S. flexneri and S. sonnei were found to be multi-resistant. S. flexneri strains were resistant to ampicillin in 83.3 percent of cases, chloramphenicol in 70.0 percent, streptomycin in 86.7 percent, sulfamethoxazole in 80.0 percent, and tetracycline in 80.0 percent, while a smaller number of strains were resistant to cephalothin (3.3 percent) and sulfazotrim (10.0 percent). S. sonnei strains were mainly resistant to sulfamethoxazole (100.0 percent) and tetracycline (96.7 percent) and, to a lesser extent, to ampicillin (6.7 percent) and streptomycin (26.7 percent). Polymerase chain reaction-based typing supported the existence of specific clones responsible for the shigellosis cases in the different cities and there was evidence of transmission between cities. This clonal structure would probably be the result of selection for virulence and resistance phenotypes. These data indicate that the human sanitary conditions of the cities investigated should be improved.