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1.
Microb Genom ; 7(8)2021 08.
Article in English | MEDLINE | ID: mdl-34427554

ABSTRACT

Shigellosis in men who have sex with men (MSM) is caused by multidrug resistant Shigellae, exhibiting resistance to antimicrobials including azithromycin, ciprofloxacin and more recently the third-generation cephalosporins. We sequenced four blaCTX-M-27-positive MSM Shigella isolates (2018-20) using Oxford Nanopore Technologies; three S. sonnei (identified as two MSM clade 2, one MSM clade 5) and one S. flexneri 3a, to explore AMR context. All S. sonnei isolates harboured Tn7/Int2 chromosomal integrons, whereas S. flexneri 3a contained the Shigella Resistance Locus. All strains harboured IncFII pKSR100-like plasmids (67-83kbp); where present blaCTX-M-27 was located on these plasmids flanked by IS26 and IS903B, however blaCTX-M-27 was lost in S. flexneri 3a during storage between Illumina and Nanopore sequencing. IncFII AMR regions were mosaic and likely reorganised by IS26; three of the four plasmids contained azithromycin-resistance genes erm(B) and mph(A) and one harboured the pKSR100 integron. Additionally, all S. sonnei isolates possessed a large IncB/O/K/Z plasmid, two of which carried aph(3')-Ib/aph(6)-Id/sul2 and tet(A). Monitoring the transmission of mobile genetic elements with co-located AMR determinants is necessary to inform empirical treatment guidance and clinical management of MSM-associated shigellosis.


Subject(s)
Bacterial Proteins/genetics , Dysentery, Bacillary/transmission , Homosexuality, Male , Plasmids/genetics , Sexual and Gender Minorities , Shigella/genetics , beta-Lactamases/genetics , Adult , Anti-Bacterial Agents/therapeutic use , DNA, Bacterial , Dysentery, Bacillary/drug therapy , Humans , Male , Middle Aged , Nanopores , Shigella/classification , Shigella sonnei/genetics , Shigella sonnei/isolation & purification , United Kingdom , Virulence/genetics , Young Adult
2.
Sci Rep ; 11(1): 12738, 2021 06 17.
Article in English | MEDLINE | ID: mdl-34140589

ABSTRACT

Both Campylobacter- and Shigella-induced invasive enteritis are common in under-5 Bangladeshi children. Our study aimed to determine the factors associated with Campylobacter and Shigella enteritis among under-5 children, the post-infection worsening growth, and the household cost of invasive enteritis. Data of children having Shigella (591/803) and Campylobacter (246/1148) isolated from the fecal specimen in Bangladesh were extracted from the Global Enteric Multicenter Study (GEMS) for the period December 2007 to March 2011. In multiple logistic regression analysis, fever was observed more frequently among shigellosis cases [adjusted OR 2.21; (95% CI 1.58, 3.09)]. Breastfeeding [aOR 0.55; (95% CI 0.37, 0.81)] was found to be protective against Shigella. The generalized estimating equations multivariable model identified a negative association between Shigella and weight-for-height z score [aOR - 0.11; (95% CI - 0.21, - 0.001)]; a positive association between symptomatic Campylobacter and weight-for-age z score [aOR 0.22; (95% CI 0.06, 0.37)] and weight-for-height z score [aOR 0.22; (95% CI 0.08, 0.37)]. Total costs incurred by households were more in shigellosis children than Campylobacter-induced enteritis ($4.27 vs. $3.49). Households with low-level maternal education tended to incur less cost in case of their shigellosis children. Our findings underscore the need for preventive strategies targeting Shigella infection, which could potentially reduce the disease burden, associated household costs, and child growth faltering.


Subject(s)
Cost of Illness , Enteritis/epidemiology , Growth , Bangladesh/epidemiology , Campylobacter jejuni/isolation & purification , Child, Preschool , Enteritis/economics , Enteritis/microbiology , Feces/microbiology , Humans , Risk Factors , Severity of Illness Index , Shigella flexneri/isolation & purification , Shigella sonnei/isolation & purification
3.
Eur J Clin Microbiol Infect Dis ; 40(7): 1573-1577, 2021 Jul.
Article in English | MEDLINE | ID: mdl-33569724

ABSTRACT

In 2019, an outbreak of Shigella sonnei occurred during two youth camps in Belgium. The clustering of isolates from both camps was confirmed by next-generation sequencing, as well as a secondary infection of a technician. The outbreak strain clustered with internationally isolated strains from patients with recent travel history to Central America. This report exemplifies enhanced surveillance and international collaboration between public health institutes by enabling to link local outbreaks to region-specific sublineages circulating abroad.


Subject(s)
Disease Outbreaks , Dysentery, Bacillary/epidemiology , Dysentery, Bacillary/microbiology , Shigella sonnei/genetics , Adult , Belgium/epidemiology , Central America , Child , Feces/microbiology , Humans , Phylogeny , Shigella sonnei/isolation & purification
5.
J Microbiol Biotechnol ; 30(10): 1516-1524, 2020 Oct 28.
Article in English | MEDLINE | ID: mdl-32807755

ABSTRACT

Climate change is expected to affect not only availability and quality of water, the valuable resource of human life on Earth, but also ultimately public health issue. A six-year monitoring (total 20 times) of Escherichia coli O157, Salmonella enterica, Legionella pneumophila, Shigella sonnei, Campylobacter jejuni, and Vibrio cholerae was conducted at five raw water sampling sites including two lakes, Hyundo region (Geum River) and two locations near Water Intake Plants of Han River (Guui region) and Nakdong River (Moolgeum region). A total 100 samples of 40 L water were tested. Most of the targeted bacteria were found in 77% of the samples and at least one of the target bacteria was detected (65%). Among all the detected bacteria, E. coli O157 were the most prevalent with a detection frequency of 22%, while S. sonnei was the least prevalent with a detection frequency of 2%. Nearly all the bacteria (except for S. sonnei) were present in samples from Lake Soyang, Lake Juam, and the Moolgeum region in Nakdong River, while C. jejuni was detected in those from the Guui region in Han River. During the six-year sampling period, individual targeted noxious bacteria in water samples exhibited seasonal patterns in their occurrence that were different from the indicator bacteria levels in the water samples. The fact that they were detected in the five Korea's representative water environments make it necessary to establish the chemical and biological analysis for noxious bacteria and sophisticated management systems in response to climate change.


Subject(s)
Water Microbiology , Water Resources , Campylobacter jejuni/isolation & purification , Campylobacter jejuni/metabolism , Chemical Phenomena , Escherichia coli O157/isolation & purification , Escherichia coli O157/metabolism , Legionella pneumophila/isolation & purification , Legionella pneumophila/metabolism , Republic of Korea , Rivers/microbiology , Salmonella enterica/isolation & purification , Salmonella enterica/metabolism , Shigella sonnei/isolation & purification , Shigella sonnei/metabolism , Vibrio cholerae/isolation & purification , Vibrio cholerae/metabolism , Water Quality
7.
Am J Public Health ; 110(6): 842-849, 2020 06.
Article in English | MEDLINE | ID: mdl-32298181

ABSTRACT

Objectives. To investigate a shigellosis outbreak in Genesee County, Michigan (including the City of Flint), and Saginaw County, Michigan, in 2016 and address community concerns about the role of the Flint water system.Methods. We met frequently with community members to understand concerns and develop the investigation. We surveyed households affected by the outbreak, analyzed Shigella isolate data, examined the geospatial distribution of cases, and reviewed available water quality data.Results. We surveyed 83 households containing 158 cases; median age was 10 years. Index case-patients from 55 of 83 households (66%) reported contact with a person outside their household who wore diapers or who had diarrhea in the week before becoming ill; results were similar regardless of household drinking water source. Genomic diversity was not consistent with a point source. In Flint, no space-time clustering was identified, and average free chlorine residual values remained above recommended levels throughout the outbreak period.Conclusions. The outbreak was most likely caused by person-to-person contact and not by the Flint water system. Consistent community engagement was essential to the design and implementation of the investigation.


Subject(s)
Disease Outbreaks/statistics & numerical data , Drinking Water/microbiology , Dysentery, Bacillary , Shigella sonnei , Water Supply , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Cities , Dysentery, Bacillary/epidemiology , Dysentery, Bacillary/microbiology , Dysentery, Bacillary/transmission , Female , Humans , Infant , Male , Michigan/epidemiology , Middle Aged , Shigella sonnei/classification , Shigella sonnei/genetics , Shigella sonnei/isolation & purification , Water Quality , Young Adult
9.
Zhonghua Liu Xing Bing Xue Za Zhi ; 40(8): 988-991, 2019 Aug 10.
Article in Chinese | MEDLINE | ID: mdl-31484266

ABSTRACT

Objective: To investigate the etiologic and epidemiologic features of an infectious diarrhea outbreak in a boarding school in Fuyang city, Anhui province. Methods: Traceability hypothesis of this study was tested according to the epidemiological characteristics of the cases. Feces, anal swabs, water samples and food residues related to the patients and chefs were collected for pathogen isolation and detection. Biochemical identification, virulence gene detection, drug susceptibility test, PFGE and multilocus sequence typing were performed. Results: The incidence rate (3.41%) of different dormitory buildings within the water supply area by shallow wells was higher than that (0.98%) of the deep wells, with statistical significance (χ(2)=17.215, P<0.001). Sixteen strains belonged to the Shigella Sonneri family were isolated from the patient's samples, and all carrying the ipaH gene. Seven strains belonged to sen and ial genes. Set1 gene that did not appear in all the 16 strains were highly resistant to ampicillin, tetracycline, compound xinnomine, cefazoline, cefotaxime, gentamicin, naphthidinic acid and streptomycin, including 9 strains to doxycycline. The pulse field pattern of the 16 strains of Shigella sonneri appeared the same, with the ST type as ST152. Conclusion: When combined data from the etiological and epidemiological investigation, it was confirmed that Shigella sonneri was the pathogen of this outbreak, and water from the shallow wells might be responsible for the source of infection.


Subject(s)
Disease Outbreaks , Dysentery, Bacillary/diagnosis , Dysentery, Bacillary/epidemiology , Feces/microbiology , Adolescent , Anti-Bacterial Agents/therapeutic use , China/epidemiology , Dysentery, Bacillary/drug therapy , Dysentery, Bacillary/microbiology , Female , Humans , Incidence , Male , Microbial Sensitivity Tests , Shigella sonnei/drug effects , Shigella sonnei/isolation & purification
10.
J Glob Antimicrob Resist ; 18: 298-303, 2019 09.
Article in English | MEDLINE | ID: mdl-31376513

ABSTRACT

BACKGROUND: Multidrug-resistant Shigella isolates have recently emerged as a serious public health threat worldwide. In particular, overseas travel is a risk factor for acquisition of antimicrobial-resistant Shigella strains. To explore the role of travel in the spread of cefotaxime-resistant Shigella sonnei in Korea, we screened 751 Shigella spp. isolates from 2007 to 2016 through the National Surveillance system, and 28 cephalosporin-resistant S. sonnei isolates were identified. METHODS: For cephalosporin-resistant S. sonnei isolates, epidemiological and molecular analyses (plasmid structure analysis, pulsed-field gel electrophoresis (PFGE) and high-quality single-nucleotide polymorphisms (hqSNPs) based on whole-genome sequencing (WGS)) were conducted to investigate the source of infection and transmission route. RESULTS: Among the 28 cefotaxime-resistant S. sonnei strains, 18 were isolated from travellers returning from Asia, including Vietnam (n=11). Molecular analysis of 18 blaCTX-M-type isolates revealed that 15 contain CTX-M-15; 50% of isolates from domestic patients contain CTX-M-14. Analysis of the genetic environments of the blaCTX-M-14 and blaCTX-M-15 genes revealed different genetic organization surrounding the blaCTX-M genes. Additionally, PFGE and hqSNP results suggested a large phylogenetic distance between the S. sonnei isolates related to overseas travel and those acquired domestically in Korea. CONCLUSION: Our study data demonstrates that two prevalent blaCTX-M genes, blaCTX-M-14 and blaCTX-M-15, have been circulating in S. sonnei in Korea over the last 10 years. Recently, international travellers are at a high risk for acquisition of CTX-M-15-producing S. sonnei in Korea.


Subject(s)
Shigella sonnei/enzymology , Shigella sonnei/genetics , Travel , beta-Lactamases/genetics , Anti-Bacterial Agents/pharmacology , Asia , Drug Resistance, Multiple, Bacterial/genetics , Dysentery/microbiology , Electrophoresis, Gel, Pulsed-Field , Humans , Microbial Sensitivity Tests , Molecular Epidemiology , Prevalence , Republic of Korea/epidemiology , Risk Factors , Shigella sonnei/drug effects , Shigella sonnei/isolation & purification , Vietnam , Whole Genome Sequencing
11.
Microb Drug Resist ; 25(10): 1465-1474, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31369341

ABSTRACT

Background: Emergence of multidrug-resistant Shigella, a major causative agent of bacterial dysentery, has generated many concerns not only in China but also worldwide. However, the prevalence of Shigella resistance caused by integron in the nonpopular season of diarrhea is not clear. Materials and Methods: Thirty-one Shigella flexneri and 22 Shigella sonnei samples collected in December 2010 from 10 cities of China were characterized for antimicrobial susceptibility, gene cassettes, widespread of integrons, and pulsed-field gel electrophoresis (PFGE) profile. Results: Multidrug resistance (MDR) was detected in 29 (93.5%) S. flexneri and 20 (90.9%) S. sonnei isolates. Class 1 integrons were detected in 25 (80.6%) S. flexneri and in 13 (59.1%) S. sonnei isolates; class 2 integrons were detected in 26 (83.9%) S. flexneri and in 19 (86.4%) S. sonnei isolates. Interestingly, the atypical class 1 integrons were mostly detected in S. flexneri (45.2%) isolates, whereas in only 1 (4.5%) S. sonnei isolate. DNA sequencing revealed two novel cassette arrays, dfrA5 and aacA4-cmlA, of class 1 integrons in S. flexneri, and dfrA17-aadA5 in S. sonnei isolates. The cassette arrays, dfrA1-sat1-aadA1 of class 2 integron and blaoxa-30-aadA1 of atypical class 1 integron, were also identified. PFGE profiles demonstrated A6 subtype of S. flexneri strains prevalent in Shanghai, Changchun, Jinan, and Changsha; and F6 subtype of S. sonnei prevalent in Jinan, Changchun, and Shanghai. Conclusion: The dissemination of MDR Shigella strains with integrons makes it an increasing public health problem in China. Increased surveillance and the development of adequate prevention strategies are warranted.


Subject(s)
Anti-Bacterial Agents/pharmacology , Dysentery, Bacillary/epidemiology , Integrons/genetics , Shigella flexneri/drug effects , Shigella sonnei/drug effects , China/epidemiology , Drug Resistance, Multiple, Bacterial/genetics , Dysentery, Bacillary/microbiology , Electrophoresis, Gel, Pulsed-Field , Humans , Sequence Analysis, DNA , Shigella flexneri/genetics , Shigella flexneri/isolation & purification , Shigella sonnei/genetics , Shigella sonnei/isolation & purification
13.
Microb Genom ; 5(5)2019 05.
Article in English | MEDLINE | ID: mdl-31099740

ABSTRACT

Shigella species are a major cause of gastroenteritis worldwide, and Shigella sonnei is the most common species isolated within the United States. Previous surveillance work in Pennsylvania documented increased antimicrobial resistance (AMR) in S. sonnei associated with reported illnesses. The present study examined a subset of these isolates by whole genome sequencing (WGS) to determine the relationship between domestic and international isolates, to identify genes that may be useful for identifying specific Global Lineages of S. sonnei and to test the accuracy of WGS for predicting AMR phenotype. A collection of 22 antimicrobial-resistant isolates from patients infected within the United States or while travelling internationally between 2009 and 2014 was chosen for WGS. Phylogenetic analysis revealed both international and domestic isolates were one of two previously defined Global Lineages of S. sonnei, designated Lineage II and Lineage III. Twelve of 17 alleles tested distinguish these two lineages. Lastly, genome analysis was used to identify AMR determinants. Genotypic analysis was concordant with phenotypic resistance for six of eight antibiotic classes. For aminoglycosides and trimethoprim, resistance genes were identified in two and three phenotypically sensitive isolates, respectively. This article contains data hosted by Microreact.


Subject(s)
Drug Resistance, Multiple, Bacterial/genetics , Dysentery, Bacillary/microbiology , Phylogeny , Shigella sonnei/classification , Shigella sonnei/genetics , Whole Genome Sequencing , Alleles , Anti-Bacterial Agents/pharmacology , DNA, Bacterial/genetics , Dysentery, Bacillary/epidemiology , Genome, Bacterial , Genotype , Humans , Microbial Sensitivity Tests , Phenotype , Polymorphism, Single Nucleotide , Sequence Analysis, DNA , Shigella sonnei/drug effects , Shigella sonnei/isolation & purification , United States/epidemiology
14.
J Glob Antimicrob Resist ; 19: 122-128, 2019 12.
Article in English | MEDLINE | ID: mdl-31077861

ABSTRACT

OBJECTIVES: Shigella spp. are an important group of waterborne pathogens worldwide. This study aimed to determine the frequency of Shigella spp. in a large collection of water samples and to uncover molecular aspects of antimicrobial resistance in the recovered isolates. METHODS: The antimicrobial resistance patterns, antimicrobial resistance genes (ARGs), including ß-lactamases (blaTEM, blaSHV, blaCTX-M, blaOXA, blaPER, blaVEB, blaGES and blaCMY), carbapenemases (blaKPC, blaNDM and blaIMP), plasmid-mediated quinolone resistance (PMQR) genes [qnrA, qnrB, qnrS and aac(6')-Ib] and tetracycline resistance genes [tet(A), tet(B), tet(C) and tet(D)], as well as class 1 and 2 integrons were analysed in Shigella spp. isolated from different water sources in Iran. RESULTS: Of 788 tested samples, Shigella sonnei and Shigella flexneri were detected in 9 (1.1%) and 6 (0.8%) samples, respectively. A multidrug-resistant (MDR) phenotype was observed in all of the isolates. Among the 15 Shigella isolates, 12 (80.0%), 5 (33.3%) and 7 (46.7%) were positive for genes encoding ß-lactam resistance, PMQR and tetracycline resistance, respectively. Class 1 integrons were more frequently detected among the isolates (8/15; 53.3%), consisting of 7 isolates (87.5%) with dfrA17-aadA5 and 1 isolate (12.5%) with sat1-aadA1 gene cassettes. The class 2 integron was detected in 3 isolates (20.0%) with the classic gene cassette array dfrA1-sat2-aadA1. CONCLUSIONS: Overall, this study showed that Shigella spp. are prevalent in water sources in Iran. Furthermore, the potential role of ARGs and integrons in the emergence of a MDR phenotype in Shigella isolates of water origin was demonstrated.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Drug Resistance, Multiple, Bacterial , Integrons , Shigella/classification , Prevalence , Quinolones/pharmacology , Sequence Analysis, DNA/methods , Shigella/drug effects , Shigella/genetics , Shigella/isolation & purification , Shigella flexneri/drug effects , Shigella flexneri/genetics , Shigella flexneri/isolation & purification , Shigella sonnei/drug effects , Shigella sonnei/genetics , Shigella sonnei/isolation & purification , Tetracyclines/pharmacology , Water , Water Microbiology , beta-Lactamases/genetics
15.
Int J Infect Dis ; 82: 117-123, 2019 May.
Article in English | MEDLINE | ID: mdl-30831222

ABSTRACT

OBJECTIVES: Ultraorthodox Jewish populations living in towns with good sanitary infrastructure but with conditions of crowding have been the epicenter of Shigella sonnei shigellosis outbreaks. In this study, the incidence and risk factors of S. sonnei shigellosis in children living in an ultraorthodox community were determined. METHODS: Data for the years 2000-2013 for all reported culture-proven S. sonnei shigellosis cases in children aged 0-59 months in the city of Elad were compared with data for the rest of the sub-district. Environmental factors obtained through parental interviews were evaluated for 78 incident cases of S. sonnei shigellosis and 141 community controls, matched by age, sex, and neighborhood. Conditional logistic regression models were performed. RESULTS: Cyclic epidemics of S. sonnei shigellosis occurred every 2 years. The mean annual incidence was 10.0 per 1000 children in Elad (95% confidence interval 7.9-12.6) vs. 3.8 per 1000 children (95% confidence interval 3.3-4.4) in the sub-district (p<0.001). Concurrent diarrheal disease in family members, having the same person in the daycare center responsible for food handling and changing diapers, and more rooms and sinks in the center, were positively associated with S. sonnei shigellosis, while children's hand-washing before meals was inversely associated. CONCLUSIONS: The burden of S. sonnei shigellosis in ultraorthodox communities is high. Enhanced hygiene interventions are required for epidemic control.


Subject(s)
Diarrhea/epidemiology , Disease Outbreaks/prevention & control , Dysentery, Bacillary/epidemiology , Epidemics , Shigella sonnei/isolation & purification , Child, Preschool , Diarrhea/microbiology , Diarrhea/prevention & control , Dysentery, Bacillary/microbiology , Dysentery, Bacillary/prevention & control , Female , Humans , Hygiene , Incidence , Infant , Infant, Newborn , Israel/epidemiology , Logistic Models , Male , Risk Factors
17.
Zhonghua Liu Xing Bing Xue Za Zhi ; 40(2): 165-169, 2019 Feb 10.
Article in Chinese | MEDLINE | ID: mdl-30744266

ABSTRACT

Objective: To analyze the pathogenic surveillance programs and related factors on bacillary dysentery in Beijing, 2008-2017, to provide evidence for the practices of diagnosis, treatment and prevention of the disease. Methods: Analysis was conducted on surveillance data of bacillary dysentery, collected from the surveillance areas of national bacillary dysentery in Beijing. Shigella positive rate of stool samples were used as the gold standard while detection rate of Shigella, diagnostic accordance rate and resistance were computed on data from the surveillance programs. Chi-square test was used to compare the rates and unconditional logistic regression was used to analyze the related factors of Shigella infection. Results: Both the reported incidence rate on bacillary dysentery and detection rate of Shigella in diarrhea patients showed significantly decreasing trend, from 2008 to 2017. The accordance rate of bacillary dysentery was only 7.80% (111/1 423). Shigella sonnei was the most frequently isolated strain (73.95%, 159/215) followed by Shigella flexnery. Results from the multivariate logistic regression of Shigella positive rate revealed that among those patients who were routine test of stool positive vs. routine test of stool positive (OR=1.863, 95%CI: 1.402-2.475), onset from July to October vs. other months'time (OR=7.271, 95%CI: 4.514-11.709) temperature ≥38 ℃vs. temperature <38 ℃(OR=4.516, 95%CI: 3.369-6.053) and age from 6 to 59 years old vs. other ages (OR=1.617, 95%CI: 1.085-2.410), presenting higher positive detection rates of Shigella from the stool tests. The resistant rates on ampicillin and nalidixic acid were 97.57% (201/206) and 94.90% (186/196), both higher than on other antibiotics. The resistant rates on ciprofloxacin (16.33%, 32/196), ofloxacin (9.57%, 11/115) and on amoxilin (15.05%, 31/206) were relatively low. The resistant rate appeared higher on Shigella flexnery than on Shigella sonnei. The proportion of strains with resistance on 3 more drugs, was 30.00%(21/70). Conclusions: The diagnostic accordance rate of bacillary dysentery in Beijing was low, with severe resistance of Shigella. Our findings suggested that clinicians should take multiple factors into account in their practices about epidemiological history, clinical symptom and testing results for diarrhea patients.


Subject(s)
Dysentery, Bacillary/diagnosis , Dysentery, Bacillary/epidemiology , Feces/microbiology , Population Surveillance/methods , Shigella/isolation & purification , Adolescent , Adult , Anti-Bacterial Agents/therapeutic use , Beijing/epidemiology , Child , China/epidemiology , Dysentery, Bacillary/drug therapy , Dysentery, Bacillary/prevention & control , Humans , Middle Aged , Sentinel Surveillance , Shigella/classification , Shigella flexneri/isolation & purification , Shigella sonnei/isolation & purification , Young Adult
20.
J Biotechnol ; 287: 1-7, 2018 Dec 10.
Article in English | MEDLINE | ID: mdl-30261194

ABSTRACT

This study provided a system for bacteria detection based on a lysosome-like-vacuole response in the yeast Saccharomyces cerevisiae. Vacuoles are factors known to activate the immune system in the presence of foreign substances. Here, Shigella sonnei and Shigella flexneri were exposed to yeast to analyze the alteration of vacuolar enzymes. The ability to detect the bacteria was evaluated by confocal microscopy after exposing and staining vacuoles with LysoTracker. Results showed that the treatment of yeast with these bacteria increased the number of red vacuole-like organelles surrounding yeast nuclei. Thus, vacuole alteration can be used as a biomarker for bacteria detection. Next, the expression of vacuolar enzymes under the influence of bacteria was examined using two-dimensional gel electrophoresis (2-DE) method for screening specific biomarkers for each Shigella strain. Finally, the recombinant yeasts that contained biomarkers fused to different fluorescent proteins confirmed the ability of yeast to detect these two Shigella strains at concentrations ranging from 10 to 100 CFU/mL.


Subject(s)
Bacterial Typing Techniques/methods , Saccharomyces cerevisiae/cytology , Shigella flexneri/isolation & purification , Shigella sonnei/isolation & purification , Vacuoles , Amines , Bacterial Proteins/analysis , Bacterial Proteins/metabolism , Biomarkers , Colony Count, Microbial , Electrophoresis, Gel, Two-Dimensional , Fluorescent Dyes , Microscopy, Confocal , Saccharomyces cerevisiae/enzymology , Saccharomyces cerevisiae/metabolism , Shigella flexneri/chemistry , Shigella flexneri/classification , Shigella sonnei/chemistry , Shigella sonnei/classification , Vacuoles/chemistry , Vacuoles/enzymology , Vacuoles/microbiology
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