Characterization of commercially formulated microbial products using molecular & biochemical methods of analysis

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Garrett, Jason Winsor
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University of Guelph
Abstract

Commercially formulated products containing bacteria cultures as their active ingredient, referred to as "Bioproducts", are available to the public throughout Canada for various domestic applications. Bioproducts are difficult for governments to regulate as manufacturers often do not provide sufficient information of their composition. This study attempted to provide regulators with useful research tools to better screen uncharacterized bioproducts by examining Biotize and Cycle. Both are liquid formulations used as biological amendments for aquariums. Investigations used culture-based methods with fatty acid methyl ester (FAME) analysis, denaturing gradient gel electrophoresis (DGGE), real-time polymerase chain reaction (PCR) and DNA-microarray analysis, to characterize microbial composition. All bacteria indicated on the manufacturer's label were identified for Biotize, but inappropriate culture conditions failed to isolate de-nitrifying bacteria in Cycle. Both bioproducts contained bacterial species not listed on the manufacturers' label. For Cycle these additional bacterial specks included three 'Mycobacterium spp'. and ' Pseudomonas aeruginosa', all of which are opportunistic pathogens. Single antibiotic and multidrug resistance were observed in both bioproducts. Real-time PCR with SYBR Green failed to detect 'Campylobacter jejuni, Escherichia coli' O157:H7, 'Listeria monocytogenes ' or 'Salmonella enterica' in either bioproduct.

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Keywords
commercially formulated product, bacteria cultures, Bioproducts, regulation, Biotize, Cycle, microbial composition
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