Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application Techniques

The quantification of Clostridia in animal feed (grass and silage) and in samples relevant for animal feed production (soil and manure) using culture methods is generally challenging, as no single culture medium is exclusively selective for all Clostridium species while inhibiting other microorganis...

Full description

Saved in:
Bibliographic Details
Main Authors: Samart Dorn-In, Hanna Geißler, Katrin Harms, Claudia Guldimann, Hubert Spiekers, Michael Diepolder, Karin Schwaiger
Format: Article
Language:English
Published: Wiley 2025-01-01
Series:International Journal of Agronomy
Online Access:http://dx.doi.org/10.1155/ioa/9930437
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850205075246415872
author Samart Dorn-In
Hanna Geißler
Katrin Harms
Claudia Guldimann
Hubert Spiekers
Michael Diepolder
Karin Schwaiger
author_facet Samart Dorn-In
Hanna Geißler
Katrin Harms
Claudia Guldimann
Hubert Spiekers
Michael Diepolder
Karin Schwaiger
author_sort Samart Dorn-In
collection DOAJ
description The quantification of Clostridia in animal feed (grass and silage) and in samples relevant for animal feed production (soil and manure) using culture methods is generally challenging, as no single culture medium is exclusively selective for all Clostridium species while inhibiting other microorganisms present in these samples. As an alternative to culture methods, this study developed a qPCR assay. It was tested for specificity with 40 clostridial and 37 other microbial species. The limit of detection of C. perfringens in each artificially contaminated sample (soil, manure, grass, and grass silage) was 100 CFU/g. The developed PCR was applied to quantify Clostridia in soil (n = 16), manure (n = 5), grass (n = 96), and grass silage (n = 80) samples that were obtained from an experimental grassland with three different manure application methods and a control group (mineral/inorganic fertilizer). Statistical analysis showed that the number of Clostridia in grass samples from the disc injector was significantly lower than that from trailing shoe and broadcast methods (p<0.05). However, the number of Clostridia in grass silage samples from the three manure application methods did not show any difference (p>0.05). The qPCR method developed in this study proved to be reliable for quantification of Clostridia in feed and environmental samples and can aid in testing and monitoring feed quality with respect to Clostridia levels.
format Article
id doaj-art-9d9cf35aa24e400db015cb53655c4275
institution OA Journals
issn 1687-8167
language English
publishDate 2025-01-01
publisher Wiley
record_format Article
series International Journal of Agronomy
spelling doaj-art-9d9cf35aa24e400db015cb53655c42752025-08-20T02:11:11ZengWileyInternational Journal of Agronomy1687-81672025-01-01202510.1155/ioa/9930437Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application TechniquesSamart Dorn-In0Hanna Geißler1Katrin Harms2Claudia Guldimann3Hubert Spiekers4Michael Diepolder5Karin Schwaiger6Unit of Food Hygiene and TechnologyChair of Food Safety and AnalyticsBavarian State Research Center for Agriculture (LfL)Chair of Food Safety and AnalyticsBavarian State Research Center for Agriculture (LfL)Bavarian State Research Center for Agriculture (LfL)Unit of Food Hygiene and TechnologyThe quantification of Clostridia in animal feed (grass and silage) and in samples relevant for animal feed production (soil and manure) using culture methods is generally challenging, as no single culture medium is exclusively selective for all Clostridium species while inhibiting other microorganisms present in these samples. As an alternative to culture methods, this study developed a qPCR assay. It was tested for specificity with 40 clostridial and 37 other microbial species. The limit of detection of C. perfringens in each artificially contaminated sample (soil, manure, grass, and grass silage) was 100 CFU/g. The developed PCR was applied to quantify Clostridia in soil (n = 16), manure (n = 5), grass (n = 96), and grass silage (n = 80) samples that were obtained from an experimental grassland with three different manure application methods and a control group (mineral/inorganic fertilizer). Statistical analysis showed that the number of Clostridia in grass samples from the disc injector was significantly lower than that from trailing shoe and broadcast methods (p<0.05). However, the number of Clostridia in grass silage samples from the three manure application methods did not show any difference (p>0.05). The qPCR method developed in this study proved to be reliable for quantification of Clostridia in feed and environmental samples and can aid in testing and monitoring feed quality with respect to Clostridia levels.http://dx.doi.org/10.1155/ioa/9930437
spellingShingle Samart Dorn-In
Hanna Geißler
Katrin Harms
Claudia Guldimann
Hubert Spiekers
Michael Diepolder
Karin Schwaiger
Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application Techniques
International Journal of Agronomy
title Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application Techniques
title_full Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application Techniques
title_fullStr Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application Techniques
title_full_unstemmed Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application Techniques
title_short Quantitative PCR Detection of Clostridia and Evaluation of Feed Hygiene Across Different Manure Application Techniques
title_sort quantitative pcr detection of clostridia and evaluation of feed hygiene across different manure application techniques
url http://dx.doi.org/10.1155/ioa/9930437
work_keys_str_mv AT samartdornin quantitativepcrdetectionofclostridiaandevaluationoffeedhygieneacrossdifferentmanureapplicationtechniques
AT hannageißler quantitativepcrdetectionofclostridiaandevaluationoffeedhygieneacrossdifferentmanureapplicationtechniques
AT katrinharms quantitativepcrdetectionofclostridiaandevaluationoffeedhygieneacrossdifferentmanureapplicationtechniques
AT claudiaguldimann quantitativepcrdetectionofclostridiaandevaluationoffeedhygieneacrossdifferentmanureapplicationtechniques
AT hubertspiekers quantitativepcrdetectionofclostridiaandevaluationoffeedhygieneacrossdifferentmanureapplicationtechniques
AT michaeldiepolder quantitativepcrdetectionofclostridiaandevaluationoffeedhygieneacrossdifferentmanureapplicationtechniques
AT karinschwaiger quantitativepcrdetectionofclostridiaandevaluationoffeedhygieneacrossdifferentmanureapplicationtechniques