Shiga Toxin 2
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Microbiology ◽  
2021 ◽  
Vol 167 (12) ◽  
Author(s):  
Takeshi Shimizu ◽  
Manami Onuki ◽  
Shin Suzuki ◽  
Shinichiro Hirai ◽  
Eiji Yokoyama ◽  
...  

Enterohaemorrhagic Escherichia coli (EHEC) produces Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2). Although stx1 and stx2 were found within the late operons of the Stx-encoding phages (Stx-phages), stx1 could mainly be transcribed from the stx1 promoter (P Stx1), which represents the functional operator-binding site (Fur box) for the transcriptional regulator Fur (ferric uptake regulator), upstream of stx1. In this study, we found that the production of Stx1 by EHEC was affected by oxygen concentration. Increased Stx1 production in the presence of oxygen is dependent on Fur, which is an Fe2+-responsive transcription factor. The intracellular Fe2+ pool was lower under microaerobic conditions than under anaerobic conditions, suggesting that lower Fe2+ availability drove the formation of less Fe2+-Fur, less DNA binding to the P Stx1 region, and an increase in Stx1 production.


2021 ◽  
Vol 9 (11) ◽  
pp. 2374
Author(s):  
Xiangning Bai ◽  
Flemming Scheutz ◽  
Henrik Mellström Dahlgren ◽  
Ingela Hedenström ◽  
Cecilia Jernberg

Shiga toxin (Stx) is the key virulence factor in the Shiga Toxin-Producing Escherichia coli (STEC), which can cause diarrhea and hemorrhagic colitis with potential life-threatening complications. There are two major types of Stx: Stx1 and Stx2. Several Stx1/Stx2 subtypes have been identified in E. coli, varying in sequences, toxicity and host specificity. Here, we report a novel Stx2 subtype (designated Stx2m) from three clinical E. coli strains isolated from diarrheal patients and asymptomatic carriers in Sweden and Denmark. The Stx2m toxin was functional and exhibited cytotoxicity in vitro. The two Swedish Stx2m-producing strains belonged to the same serotype O148:H39 and Multilocus Sequencing Typing (MLST) Sequence Type (ST) 5825, while the Danish strain belonged to the O96:H19 serotype and ST99 type. Whole-genome sequencing (WGS) data analysis revealed that the three Stx2m-producing strains harbored additional virulence genes and the macrolide resistance gene mdf (A). Our findings expand the pool of Stx2 subtypes and highlight the clinical significance of emerging STEC variants. Given the clinical relevance of the Stx2m-producing strains, we propose to include Stx2m in epidemiological surveillance of STEC infections and clinical diagnosis.


Toxins ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 775
Author(s):  
Yugyeong Lee ◽  
Min-Hyeok Kim ◽  
David Rodrigues Alves ◽  
Sejoong Kim ◽  
Luke P. Lee ◽  
...  

Shiga toxin-producing Escherichia coli (STEC) infects humans by colonizing the large intestine, and causes kidney damage by secreting Shiga toxins (Stxs). The increased secretion of Shiga toxin 2 (Stx2) by some antibiotics, such as ciprofloxacin (CIP), increases the risk of hemolytic–uremic syndrome (HUS), which can be life-threatening. However, previous studies evaluating this relationship have been conflicting, owing to the low frequency of EHEC infection, very small number of patients, and lack of an appropriate animal model. In this study, we developed gut–kidney axis (GKA) on chip for co-culturing gut (Caco-2) and kidney (HKC-8) cells, and observed both STEC O157:H7 (O157) infection and Stx intoxication in the gut and kidney cells on the chip, respectively. Without any antibiotic treatment, O157 killed both gut and kidney cells in GKA on the chip. CIP treatment reduced O157 infection in the gut cells, but increased Stx2-induced damage in the kidney cells, whereas the gentamycin treatment reduced both O157 infection in the gut cells and Stx2-induced damage in the kidney cells. This is the first report to recapitulate a clinically relevant situation, i.e., that CIP treatment causes more damage than gentamicin treatment. These results suggest that GKA on chip is very useful for simultaneous observation of O157 infections and Stx2 poisoning in gut and kidney cells, making it suitable for studying the effects of antibiotics on the risk of HUS.


2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 339-340
Author(s):  
Zhe Pan ◽  
Yanhong Chen ◽  
Tim A McAllister

Abstract This study aimed to identify whether microbial interactions in the rectum contribute to Shiga toxin producing bacteria colonization. In total, 12 rectal digesta samples based on the previously identified Shiga toxin 2 gene (stx2) abundance (DNA) and expression (RNA) in Shiga toxin-producing bacteria (Stx2- group: detectable DNA, n=6; Stx2+ group: detectable DNA and RNA, n = 6) were subjected to microbial profiling using amplicon sequencing. Firmicutes (72.7 ± 2.0 %) and Bacteroidetes (24.6 ± 1.9 %) are the most predominant phyla of rectal microbiota, and no compositional differences were identified between two groups at the phylum level. The Shannon and Chao1 indices weren’t different in rectal digesta microbial communities between two groups. Twenty-four and thirteen taxa were identified to be group-specific genera in microbial communities from Stx2- and Stx2+ group, respectively (2 out of 6, average relative abundance >0.1%). The network analysis indicated 12 and 14 keystone taxa (Generalists, densely connected with other taxa) in microbial communities between Stx2- and Stx2+ groups, respectively. Eight out of 12 and six out of 14 generalists in the Stx2- and Stx2+ group are belonging to group-specific genera, respectively. Generalists belonging to group-specific genera were broadly distributed in Stx2- network while centralized distributed in the Stx2+ network, suggesting the higher stability of the Stx2- network structure in comparison of Stx2+ network computed by the natural connectivity measurement. However, 66 core genera shared by microbial communities between two groups were not classified into network generalists. Overall, our results indicate microbial crosstalks and keystone taxa in microbial communities between two groups differed, suggesting that the microbial interactions rather than the shifts in taxa abundance may be more important affecting host. Moreover, group-specific genera play a vital ecological role in the microbial interactions, indicating the potential for being microbial markers to differentiate Shiga toxin-producing bacteria colonization in beef cattle.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Patricia B. Lodato

AbstractEnterohaemorrhagic Escherichia coli (EHEC) comprise a group of intestinal pathogens responsible for a range of illnesses, including kidney failure and neurological compromise. EHEC produce critical virulence factors, Shiga toxin (Stx) 1 or 2, and the synthesis of Stx2 is associated with worse disease manifestations. Infected patients only receive supportive treatment because some conventional antibiotics enable toxin production. Shiga toxin 2 genes (stx2) are carried in λ-like bacteriophages (stx2-phages) inserted into the EHEC genome as prophages. Factors that cause DNA damage induce the lytic cycle of stx2-phages, leading to Stx2 production. The phage Q protein is critical for transcription antitermination of stx2 and phage lytic genes. This study reports that deficiency of two endoribonucleases (RNases), E and G, significantly delayed cell lysis and impaired production of both Stx2 and stx2-phages, unlike deficiency of either enzyme alone. Moreover, scarcity of both enzymes reduced the concentrations of Q and stx2 transcripts and slowed cell growth.


PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11871
Author(s):  
Keiji Nakamura ◽  
Chikashi Tokuda ◽  
Hideyuki Arimitsu ◽  
Yoshiki Etoh ◽  
Mitsuhiro Hamasaki ◽  
...  

Shiga toxin-producing Escherichia coli (STEC) is a major intestinal pathogen and causes serious gastrointestinal illness, which includes diarrhea, hemorrhagic colitis, and life-threatening hemolytic uremic syndrome. The major virulence factors of STEC are Shiga toxins (Stx1 and Stx2), which belong to the AB-type toxin family. Among several subtypes of Stx1 and Stx2, the production of Stx2a is thought to be a risk factor for severe STEC infections, but Stx2a production levels vary markedly between STEC strains, even strains with the same serotype. Therefore, quantitative analyses of Stx2 production by STEC strains are important to understand the virulence potential of specific lineages or sublineages. In this study, we developed a novel Stx2 quantification method by utilizing homogeneous time-resolved fluorescence resonance energy transfer (HTRF) technology. To determine suitable “sandwich” assay conditions, we tested 6 combinations of fluorescence-labeled monoclonal antibodies (mAbs) specific to Stx2 and compared the HTRF signal intensities obtained at various incubation times. Through this analysis, we selected the most suitable mAb pair, one recognizing the A subunit and the other recognizing the B subunit, thus together detecting Stx holotoxins. The optimal incubation time was also determined (18 h). Then, we optimized the concentrations of the two mAbs based on the range for linearity. The established HTRF assay detected 0.5 ng/ml of the highly purified recombinant Stx2a and Stx2e proteins and the working range was 1–64 ng/ml for both Stx2a and Stx2e. Through the quantification analysis of Stx proteins in STEC cell lysates, we confirmed that other Stx2 subtypes (Stx2b, Stx2c, Stx2d and Stx2g) can also be quantified at a certain level of accuracy, while this assay system does not detect Stx2f, which is highly divergent in sequence from other Stx2 subtypes, and Stx1. As the HTRF protocol we established is simple, this assay system should prove useful for the quantitative analysis of Stx2 production levels of a large number of STEC strains.


Toxins ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 437
Author(s):  
John K. Crane ◽  
Mashal Salehi ◽  
Cassandra L. Alvarado

Several classes of non-antibiotic drugs, including psychoactive drugs, proton-pump inhibitors (PPIs), non-steroidal anti-inflammatory drugs (NSAIDs), and others, appear to have strong antimicrobial properties. We considered whether psychoactive drugs induce the SOS response in E. coli bacteria and, consequently, induce Shiga toxins in Shiga-toxigenic E. coli (STEC). We measured the induction of an SOS response using a recA-lacZ E. coli reporter strain, as RecA is an early, reliable, and quantifiable marker for activation of the SOS stress response pathway. We also measured the production and release of Shiga toxin 2 (Stx2) from a classic E. coli O157:H7 strain, derived from a food-borne outbreak due to spinach. Some, but not all, serotonin selective reuptake inhibitors (SSRIs) and antipsychotic drugs induced an SOS response. The use of SSRIs is widespread and increasing; thus, the use of these antidepressants could account for some cases of hemolytic-uremic syndrome due to STEC and is not attributable to antibiotic administration. SSRIs could have detrimental effects on the normal intestinal microbiome in humans. In addition, as SSRIs are resistant to environmental breakdown, they could have effects on microbial communities, including aquatic ecosystems, long after they have left the human body.


Author(s):  
Daiana S. Sánchez ◽  
Lilian K. Fischer Sigel ◽  
Alejandro Balestracci ◽  
Cristina Ibarra ◽  
María M. Amaral ◽  
...  

2021 ◽  
pp. 100299
Author(s):  
Katarzyna Szymczak-Kulus ◽  
Sascha Weidler ◽  
Anna Bereznicka ◽  
Krzysztof Mikolajczyk ◽  
Radoslaw Kaczmarek ◽  
...  
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