Molecular monitoring and diagnostic of contaminants from agroindustry

Abstract

The innovation in adopting molecular techniques in the traceability of agroindustrial processes, inputs and products enables a fast and precise control of restrictive factors to production, such as contaminating microorganisms from industries still in the latent or initial phase of infection. Bacterial contamination is considered one of the main factors affecting the yield of industrial fermentation processes. For example, the ethanol industry may have significant yield losses due to contamination caused by bacteria of the genera Lactobacillus, Bacillus, Acetobacter, among others, present at levels above 105 CFU/mL of must. These can enter the system loaded with water and/or molasses and remain in it as biofilms that develop in heat exchangers, dorns, pipes and dead spots, being a continuous source of contaminant inoculation. Therefore, the precise location and quantification of these restrictive factors of the chain would enable the industry to better manage processes and to act for continuous improvement. In this context, quantitative detection by qPCR (quantitative PCR or real-time PCR), a service offered by MetaQuantiON, emerges as a strong molecular diagnostic tool for practical applications in the detection and quantification highly sensitive, specific and fast of microrganisms at the genera and species level, which in many cases are not achieved by conventional microbiological analysis. The objective of this project is the development, optimization and validation of an analysis package containing assays to quantitatively detect the most important bacterial contaminants of the alcoholic fermentation process, based on the qPCR technique. With this product, ethanol plants will be able to monitor the entire process accurately, enabling quick and specific decision making that will result in input savings (antibiotics and acid) and increased yields, operating a more economically and environmentally sustainable system. Specific and applied objectives of this proposal include: to develop protocols for optimization of DNA extraction of each sample type; design primers and probes for the genera Lactobacillus, Bacillus, Acetobacter, Pseudomonas, Clostridium, Streptococcus, Enterobacter and Leuconostoc; optimize each assay for the specificity of the microorganism and for detection and quantitation limits; and validate the tests on samples of raw material (grinding and molasses), must, wine, centrifuged yeast and after treated yeast. As a result of the research project of this proposal, MetaQunatiON intends to make available to the sugar and alcohol sector molecular diagnostic tests to monitor microbial contamination. (AU)