Novel strategy of rapid typing of Shiga toxin-producing Escherichia coli using MALDI Biotyper and ClinProTools analysis

Author(s):  
Yumi Kubo ◽  
Osamu Ueda ◽  
Sawa Nagamitsu ◽  
Hachiro Yamanishi ◽  
Akihiro Nakamura ◽  
...  
2020 ◽  
Vol 8 (11) ◽  
pp. 1801
Author(s):  
Michael Bording-Jorgensen ◽  
Brendon D. Parsons ◽  
Gillian A.M. Tarr ◽  
Binal Shah-Gandhi ◽  
Colin Lloyd ◽  
...  

Shiga toxin-producing Escherichia coli (STEC) are associated with acute gastroenteritis worldwide, which induces a high economic burden on both healthcare and individuals. Culture-independent diagnostic tests (CIDT) in frontline microbiology laboratories have been implemented in Alberta since 2019. The objectives of this study were to determine the association between gene detection and culture positivity over time using STEC microbiological clearance samples and also to establish the frequency of specimen submission. Both stx genes’ amplification by real-time PCR was performed with DNA extracted from stool samples using the easyMAG system. Stools were inoculated onto chromogenic agar for culture. An association between gene detection and culture positivity was found to be independent of which stx gene was present. CIDT can provide rapid reporting with less hands-on time and technical expertise. However, culture is still important for surveillance and early cluster detection. In addition, stool submissions could be reduced from daily to every 3–5 days until a sample is negative by culture.


Horticulturae ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 68
Author(s):  
Yi-Ju Wang ◽  
Amanda J. Deering ◽  
Hye-Ji Kim

Our previous study reported that fresh produce grown in aquaponic and hydroponic systems can pose potential food safety hazards due to an accidental introduction of contaminated fish and cross-contamination between the systems. In this study, we examined the effects of plant species and age on the likelihood and level of internalization of Shiga toxin-producing Escherichia coli (STEC) in aquaponic and hydroponic systems. Four plant species, basil (Ocimum basilicum L. cv. Genovese), cilantro (Coriandrum Sativum L.), lettuce (Lactuca sativa cv. Cherokee), and kale (Brassica oleracea var. sabellica), received root damage treatment as seedlings before transplanting or mature plants at three weeks after transplanting by cutting off 1-cm tips of one-third of the roots. Enrichments and selective media were used for the isolation, and presumptive positive colonies were confirmed by PCR for the presence of stx1 gene in plant tissues, recirculating water, and fish feces collected at four weeks after transplanting. In hydroponic systems, STEC was found neither in the solution nor in the roots and leaves of all four plant species, possibly through improved sanitation and hygiene practices. However, consistent with our previous findings, STEC was found in the water, on the plant roots, and in the fish feces in aquaponic systems, even after thorough sanitation prior to the study. Regardless of plant age, STEC was internalized in the roots of all plant species when the roots were damaged, but there was no difference in the degree of internalization with STEC among plant species. STEC was present in the leaves only when seedlings received root damage treatment and were grown to maturity, indicating that root damage allows STEC to internalize in the roots within a week, but a longer period is required for STEC to internalize into the leaves. We concluded that root damage on seedlings can cause the internalization of E. coli O157:H7 in the edible parts of leafy vegetables and herbs in soilless production systems.


2021 ◽  
Vol 45 (4) ◽  
Author(s):  
María de los Ángeles Rey ◽  
Mariana Cap ◽  
Leonardo Cristian Favre ◽  
Anabel Rodríguez Racca ◽  
María José Dus Santos ◽  
...  

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 265
Author(s):  
Peter Kotsoana Montso ◽  
Caven Mguvane Mnisi ◽  
Collins Njie Ateba ◽  
Victor Mlambo

Preslaughter starvation and subacute ruminal acidosis in cattle are known to promote ruminal proliferation of atypical enteropathogenic Escherichia coli strains, thereby increasing the risk of meat and milk contamination. Using bacteriophages (henceforth called phages) to control these strains in the rumen is a potentially novel strategy. Therefore, this study evaluated the viability of phages and their efficacy in reducing E. coli O177 cells in a simulated ruminal fermentation system. Fourteen phage treatments were allocated to anaerobic serum bottles containing a grass hay substrate, buffered (pH 6.6–6.8) bovine rumen fluid, and E. coli O177 cells. The serum bottles were then incubated at 39 °C for 48 h. Phage titres quadratically increased with incubation time. Phage-induced reduction of E. coli O177 cell counts reached maximum values of 61.02–62.74% and 62.35–66.92% for single phages and phage cocktails, respectively. The highest E. coli O177 cell count reduction occurred in samples treated with vB_EcoM_366B (62.31%), vB_EcoM_3A1 (62.74%), vB_EcoMC3 (66.67%), vB_EcoMC4 (66.92%), and vB_EcoMC6 (66.42%) phages. In conclusion, lytic phages effectively reduced E. coli O177 cells under artificial rumen fermentation conditions, thus could be used as a biocontrol strategy in live cattle to reduce meat and milk contamination in abattoirs and milking parlours, respectively.


2021 ◽  
Vol 9 (3) ◽  
pp. 503
Author(s):  
Michael Bording-Jorgensen ◽  
Hannah Tyrrell ◽  
Colin Lloyd ◽  
Linda Chui

Acute gastroenteritis caused by Shiga toxin-producing Escherichia coli (STEC) affects more than 4 million individuals in Canada. Diagnostic laboratories are shifting towards culture-independent diagnostic testing; however, recovery of STEC remains an important aspect of surveillance programs. The objective of this study was to compare common broth media used for the enrichment of STEC. Clinical isolates including O157:H7 as well as non-O157 serotypes were cultured in tryptic soy (TSB), MacConkey (Mac), and Gram-negative (GN) broths and growth was compared using culture on sheep’s blood agar and real-time PCR (qPCR). In addition, a selection of the same isolates was spiked into negative stool and enriched in the same three broths, which were then evaluated using culture on CHROMagarTM STEC agar and qPCR. TSB was found to provide the optimal enrichment for growth of isolates with and without stool. The results from this study suggest that diagnostic laboratories may benefit from enriching STEC samples in TSB as a first line enrichment instead of GN or Mac.


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