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...
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| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2025-01-01
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| Series: | International Journal of Agronomy |
| Online Access: | http://dx.doi.org/10.1155/ioa/9930437 |
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| Summary: | 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. |
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| ISSN: | 1687-8167 |