Development of a new multiplex quantitative real-time polymerase chain reaction method for Clostridium butyricum, Clostridium sporogenes and Clostridium tyrobutyricum detection in cheese

Şahiner, Aslı; Çalışkan, Sennur; Halat, Ece

doi:10.1016/j.lwt.2021.112914

Cited by 8 publications

(1 citation statement)

References 14 publications

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“…Differently, in this study, the use of molecular techniques such as real-time PCR ensured a highly sensitive detection of C. tyrobutyricum , increasing the detection and resolution capacity in complex matrices such as silage. Our results showed a sensitive limit of detection (1.5 Log CFU/mL), in line with a previous study, in which a multiplex real-time PCR was developed to detect Clostridium species in cheese with a detection limit of ~ 1 Log CFU/mL ( Şahiner et al, 2022 ). In addition, data obtained in our validation study ( Arnaboldi et al, 2021 ) showed that MPN results sometimes overestimated or underestimated the real clostridia amount in the samples and that the difference between MPN and real-time PCR was related to the specificity of the molecular method only quantifying C. tyrobutyricum DNA.…”

Section: Discussionsupporting

confidence: 91%

Clostridium tyrobutyricum occurrence in silages and cattle feed: Use of molecular and simulation data to optimize predictive models

et al. 2023

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IntroductionPoor quality silage can derive from the presence of deleterious microorganisms such as clostridia. Their dissemination along the food chain, especially in milk, causes issues such as the cheese late-blowing defect, particularly triggered by Clostridium tyrobutyricum. The scope of our study was to determine the C. tyrobutyricum occurrence in three different farms across four time periods in relation to the animal diets, specifically the Total Mixed Ration (TMR), by using real-time PCR.MethodsFor this purpose, molecular-derived data were exploited to optimize a predictive model that simulated the farm conditions favoring the growth of butyric acid bacteria such as C. tyrobutyricum.ResultsOur results showed that the originally utilized predictive model strongly underestimated the growth of C. tyrobutyricum in comparison to the molecular data. At the same time, our findings uncovered an additional source of contamination in the TMR related to silage and dietary residues that represent a reservoir of microbial contamination during successive TMR preparation. Based on these findings, the optimization of the model parameters such as growth rate range and the inclusion of the residues in the model, allowed a more accurate prediction of the contamination levels. Therefore, this study revealed that proper hygiene practices such as the removal of silage and TMR residues within the farm environment is essential to control the contamination by C. tyrobutyricum and avoid food waste and economic losses.

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Section: Discussionsupporting

confidence: 91%

Clostridium tyrobutyricum occurrence in silages and cattle feed: Use of molecular and simulation data to optimize predictive models

et al. 2023

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Quantitative Detection of Late Blowing Agents C. tyrobutyricum, C. butyricum, and C. sporogenes in Traditional Turkish Cheese by Multiplex Real-Time PCR

2023

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The effects of irrigation on the survival of Clostridium sporogenes in the phyllosphere and soil environments of lettuce

Fourie,

Van Wyk,

Bezuidenhout

et al. 2024

3 Biotech

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This study aimed to address the gap in knowledge regarding the fate of foodborne pathogens within agro-ecosystems. It specifically focused on the surrogate microorganism Clostridium sporogenes, which was introduced into lettuce-producing environments via surface and spray irrigation methods, respectively. The concentration of C. sporogenes in the rhizosphere, phyllosphere, and non-rhizosphere soil was quantified by quantitative polymerase chain reaction (qPCR) over a 42-day trial. The surface irrigation method exhibited a more noticeable contamination effect on the soil environments, compared to the phyllosphere. The results indicated a noticeable increase in C. sporogenes concentrations during the initial 22 days, with a 10.4-fold rise (0.39–4.05 log copy numbers/g soil) in the rhizosphere and 1.9-fold increase (2.97–5.59 log copy numbers/g soil) in the non-rhizosphere. However, concentrations in both soil environments subsequently decreased, falling below the initial inoculum concentration by the end of the trial. In contrast, the spray irrigation method resulted in most of the contamination being localised on the lettuce phyllosphere, with a high C. sporogenes concentration of 9.09 log copy numbers/g leaves on day 0. This concentration exponentially decreased to a minimal 0.019 log copy numbers/g leaves by day 32. Although concentrations in both soil environments decreased over time, trace concentrations of C. sporogenes were detectable at the end of the trial, posing a potential hazard to the microbiological safety of postharvest produce. These findings shed light on the dynamics of C. sporogenes in agro-ecosystems and underscore the importance of irrigation practices that ensure the safety of those who consume fresh produce.

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Development of a new multiplex quantitative real-time polymerase chain reaction method for Clostridium butyricum, Clostridium sporogenes and Clostridium tyrobutyricum detection in cheese

Cited by 8 publications

References 14 publications

Clostridium tyrobutyricum occurrence in silages and cattle feed: Use of molecular and simulation data to optimize predictive models

Clostridium tyrobutyricum occurrence in silages and cattle feed: Use of molecular and simulation data to optimize predictive models

Quantitative Detection of Late Blowing Agents C. tyrobutyricum, C. butyricum, and C. sporogenes in Traditional Turkish Cheese by Multiplex Real-Time PCR

The effects of irrigation on the survival of Clostridium sporogenes in the phyllosphere and soil environments of lettuce

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