Design and validation of a real-time PCR technique for assessing the level of inclusion of fungus- and yeast-based additives in feeds.
J Microbiol Methods
; 171: 105867, 2020 04.
Article
em En
| MEDLINE
| ID: mdl-32061906
ABSTRACT
A reliable method for quantification of non-viable microbe-based nutritional and zootechnical additives introduced into feed is essential in order to ensure regulatory compliance, feed safety and product authenticity in industrial applications. In the present work, we developed a novel real-time quantitative polymerase chain reaction (qPCR) -based analysis protocol for monitoring two microbial additives in feed matrices. To evaluate the applicability of the method, pelleted wheat- and maize-based broiler chicken diets containing a non-viable phytase-producing strain of Aspergillus niger produced in solid state fermentation (150 or 300â¯g/t) and a non-viable selenium-enriched Saccharomyces cerevisiae (100 or 200â¯g/t) as model feed ingredients, were manufactured and subjected to analysis. Power analysis of the qPCR results indicated that 2 to 6 replicate feed samples were required to distinguish the product doses applied, which confirms that the microbial DNA was efficiently recovered and that potential PCR inhibitors present in the feed material were successfully removed in DNA extraction. The analysis concept described here was shown to be an accurate and sensitive tool for monitoring the inclusion levels of non-viable, unculturable microbial supplements in animal diets.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Aspergillus niger
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Saccharomyces cerevisiae
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Reação em Cadeia da Polimerase em Tempo Real
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Ração Animal
Limite:
Animals
Idioma:
En
Revista:
J Microbiol Methods
Ano de publicação:
2020
Tipo de documento:
Article
País de publicação:
HOLANDA
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HOLLAND
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NETHERLANDS
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NL
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PAISES BAJOS
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THE NETHERLANDS