scholarly journals Evaluation of PMA‐qPCR methodology to detect and quantify viable Shiga toxin‐producing Escherichia coli in beef burgers

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

2020 ◽  
Vol 83 (5) ◽  
pp. 865-873
Author(s):  
ANNA C. S. PORTO-FETT ◽  
LAURA E. SHANE ◽  
BRADLEY A. SHOYER ◽  
MANUELA OSORIA ◽  
YANGJIN JUNG ◽  
...  

ABSTRACT We evaluated high pressure processing to lower levels of Shiga toxin–producing Escherichia coli (STEC) and Listeria monocytogenes inoculated into samples of plant or beef burgers. Multistrain cocktails of STEC and L. monocytogenes were separately inoculated (∼7.0 log CFU/g) into plant burgers or ground beef. Refrigerated (i.e., 4°C) or frozen (i.e., −20°C) samples (25 g each) were subsequently exposed to 350 MPa for up to 9 or 18 min or 600 MPa for up to 4.5 or 12 min. When refrigerated plant or beef burger samples were treated at 350 MPa for up to 9 min, levels of STEC were reduced by ca. 0.7 to 1.3 log CFU/g. However, when refrigerated plant or beef burger samples were treated at 350 MPa for up to 9 min, levels of L. monocytogenes remained relatively unchanged (ca. ≤0.3-log CFU/g decrease) in plant burger samples but were reduced by ca. 0.3 to 2.0 log CFU/g in ground beef. When refrigerated plant or beef burger samples were treated at 600 MPa for up to 4.5 min, levels of STEC and L. monocytogenes were reduced by ca. 0.7 to 4.1 and ca. 0.3 to 5.6 log CFU/g, respectively. Similarly, when frozen plant and beef burger samples were treated at 350 MPa up to 18 min, reductions of ca. 1.7 to 3.6 and ca. 0.6 to 3.6 log CFU/g in STEC and L. monocytogenes numbers, respectively, were observed. Exposure of frozen plant or beef burger samples to 600 MPa for up to 12 min resulted in reductions of ca. 2.4 to 4.4 and ca. 1.8 to 3.4 log CFU/g in levels of STEC and L. monocytogenes, respectively. Via empirical observation, pressurization did not adversely affect the color of plant burger samples, whereas appreciable changes in color were observed in pressurized ground beef. These data confirm that time and pressure levels already validated for control of STEC and L. monocytogenes in ground beef will likely be equally effective toward these same pathogens in plant burgers without causing untoward effects on product color. HIGHLIGHTS



2020 ◽  
Vol 83 (3) ◽  
pp. 434-442 ◽  
Author(s):  
JOHN B. LUCHANSKY ◽  
BRADLEY A. SHOYER ◽  
YANGJIN JUNG ◽  
LAURA E. SHANE ◽  
MANUELA OSORIA ◽  
...  

ABSTRACT The viability of Shiga toxin–producing Escherichia coli (STEC), Salmonella, and Listeria monocytogenes within plant- and beef-based burgers was monitored during storage and cooking. When inoculated (ca. 3.5 log CFU/g) into 15-g portions of plant- or beef-based burgers, levels of STEC and Salmonella decreased slightly (≤0.5-log decrease) in both types of burgers when stored at 4°C, but increased ca. 2.4 and 0.8 log CFU/g, respectively, in plant-based burgers but not beef-based burgers (≤1.2-log decrease), after 21 days at 10°C. For L. monocytogenes, levels increased by ca. 1.3 and 2.6 log CFU/g in plant burgers after 21 days at 4 and 10°C, respectively, whereas pathogen levels decreased slightly (≤0.9-log decrease) in beef burgers during storage at 4 and 10°C. Regarding cooking, burgers (ca. 114 g each) were inoculated with ca. 7.0 log CFU/g STEC, Salmonella, or L. monocytogenes and cooked in a sauté pan. Cooking plant- or beef-based burgers to 62.8°C (145°F), 68.3°C (155°F), or 73.9°C (165°F) delivered reductions ranging from ca. 4.7 to 6.8 log CFU/g for STEC, ca. 4.4 to 7.0 log CFU/g for L. monocytogenes, and ca. 3.5 to 6.7 log CFU/g for Salmonella. In summary, the observation that levels of all three pathogens increased by ca. 1.0 to ca. 2.5 log CFU/g in plant-based burgers when stored at an abusive temperature (10°C) highlights the importance of proper storage (4°C) to lessen risk. However, because all three pathogens responded similarly to heat in plant-based as in beef-based burgers, well-established cooking parameters required to eliminate STEC, Salmonella, or L. monocytogenes from ground beef should be as effective for controlling cells of these same pathogens in a burger made with plant-sourced protein. HIGHLIGHTS



2014 ◽  
Vol 20 (28) ◽  
pp. 4535-4548 ◽  
Author(s):  
Jolanta Szych ◽  
Tomasz Wo.lkowicz ◽  
Roberto Ragione ◽  
Grzegorz Madajczak


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 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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