Objective
This project aims at developing biological silage additives made of live lactic acid bacteria and enzymes. These additives are intended to improve both the conservation and the nutritional value of silage. They will be targeted at two types of feed: green forage (rye grass, alfalfa) and agricultural byproducts (residues of tomatoes, grapes or beet pulp).
Novel biological silage additives are being developed using a combination of biochemical, microbiological and genetical methods including genetic engineering. These additives will be composed of a mixture of lactic acid bacteria and enzymes. A strain of Pediococcus acidilactici (G 24) has been selected out of a panel of 18 different strains. This strain was shown to display superior performances in terms of: temperature and pH optimum and range; growth rate; rate and extent of acidification; stimulation of growth of epiphytic Lactobacillus (L) plantarum in grass silage; and preservation of silage crude protein.
Deoxyribonucleic acid (DNA) probes have been developed which allow identification of strains of L plantarum by colony hybridization. In particular, a probe derived from a region of ribosomal DNA known to be evolutionary variable has been constructed to allow discrimination of the DCU 101 strain (a grass isolate from Ireland) amongst a wide range of strains. The L-LDH gene from the Bactensil strain has been cloned and sequenced. The D- and L-LDH genes from Pediococcus acidilactici have been cloned. A 2-step method of homologous recombination allowing the targeted genetic modification of specific chromosomal genes has been successfully practised on a model silage bacterium (Enterococcus faecalis) in preparation for D-LDH gene disruption experiments to be performed on L plantarum.
A strain of L amylovorus has been successfully grown in 20 l fermenters and lyophilized. This strain is ready to be tested on lucerne silage. Experiments dealing with the effects of enzymes and bacterial additives on grass, lucerne and byproducts of silages have been started.
To achieve these objectives the following approach will be used:
Enzymes will be selected for their efficiency to bring down the lower pH limit of silage.
Lactic acid bacteria will be selected according to physiological parameters (metabolic profile, lactic fermentation, lag period, synergy, etc). Candidate strains will be evaluated for their industrial aptitudes (fermentation yield, resistance to lyophilization, shelf life, etc).
Prototype silage additives will be produced at pilot scale as lyophilized preparations.
Silage fermentation will be monitored using physical, chemical and biochemical parameters (pH, L-lactic acid and D-lactic acid, crude protein, dry matter, etc).
Silage bacterial population dynamics will be studied using phenotypic and genotypic screening methods.
The rate of L-lactic acid production of Lactobacillus plantarum will be enhanced through genetic engineering of lactic dehydrogenase (LHD) genes.
Silage nutritional value will be assessed through feeding trials conducted in experimental farms.
The final product will be enzymes which release soluble carbohydrates from the cell wall and/or reserve polysaccharides in order to enhance terminal fermentation and to increase fibre digestibility, and bacteria which rapidly bring down the pH, dominate the endogenous microflora and produce large amounts of L-lactic acid in the silo. In addition, the bacteria will be adapted for use in large scale industrial production and conditioning.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences chemical sciences organic chemistry organic acids
- natural sciences biological sciences microbiology bacteriology
- medical and health sciences medical biotechnology genetic engineering
- agricultural sciences agriculture, forestry, and fisheries agriculture industrial crops fodder
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Coordinator
1348 LOUVAIN-LA-NEUVE
Belgium
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