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Commercial Success of ECLAIR Programme
AGRE-0064: Isolation of new starter cultures from traditional cheese and fermented milk |
AGRE-0064: Isolation of new starter cultures from traditional cheese and fermented milk
Science Background
Industrial production of cheese relies exclusively on the use of selected cultures that are carefully maintained and subcultured in specialized laboratories. In contrast, many cheeses and fermented milks in Southern Europe are made by natural fermentation in which the lactic acid bacteria adventitiously present in the milk grow and produce acid. Each of these may represent a unique ecological niche with its own microflora, which differs significantly from the microflora used commercially in cheese manufacture.
Objectives
The major objective was to isolate new strains of lactic acid bacteria from traditional cheese and fermented milk for possible use as industrial cultures. A second objective was to isolate strains with unusual flavour profiles that may ultimately result in the development of new fermented dairy products with different flavours. A third objective was to develop a suitable probe for distinguishing the different subspecies of lactococci and for use in isolating strains with commercially significant properties, for example nisin or ß-galactosidase products.
Significant changes and results since end of ECLAIR
The coordinators, Teagasc, developed a Lactococcus lactis bacteriocin that underwent commercial trials in 1998 resulting in an agreement with Cross VetPharm Group Ltd, Ireland. It is anticipated that this will lead to a new product for prevention of mastitis in dairy cows by 2000. National culture collections have been established and a database of 3500-4000 strains of lactic acid bacteria developed and compiled at Universita Cattolica del Sacro Cuore (UCSC), Italy. Istituto Zootecnico e Caseario per la Sardegna (IZCS), Italy have given their microbial strains to small dairy factories in Sardinia for limited production of artisanal cheeses at an industrial level. In 1998, Centro Sperimentale del Latte, Italy added a new starter culture (STAR-IDC) to their product range, containing mesophilic cocci and Streptococcus thermophilus. Nizo, the Netherlands, have carried out follow-up contract research projects, indicating a strong interest from industry. A new project involving a number of companies is being put together, to be coordinated by UCSC.
Results
At end of this ECLAIR project
More than 4000 strains of lactic acid bacteria from 31 artisanal products were isolated. More than 90% of these were characterized and their details added to a database, developed and compiled by Piacenza, Italy. Some of the biochemical properties of these strains were also measured, including tests for exopolysaccharide (EPS) and bacteriocin production. One of the lactococci produced a heat-stable, plasmid-encoded, broad-spectrum bacteriocin that was not nisin. Phage-resistance was linked with this plasmid. Probes for identifying the different genera, species and subspecies of lactic acid bacteria were developed. A course on their use was attended by participants from this project and FLAIR Programme project AGRF-0053: The selection and characterization of human probiotic strains, the process technology necessary for their incorporation into fermented foods and evaluation in selected animal models. These probes were used to distinguish between Lactococcus species, leading to the identification of a new species L. hoecklum. Techniques for profiling flavour production of different strains were also developed. Pilot scale cheese production with strains with desirable properties resulted in some cheeses that were as good if not better than traditional cheeses.
Current position
Centro Sperimentale del Latte (CSL, Italy) carried out further in-house research related to the technical aspects of production of various types of Italian cheese. In particular, strains of lactococci and Streptococcus thermophilus were investigated. In 1995, the growth (Bactometer impedance method) and effects on pH (titratable acidity) during development were investigated at the temperatures used in the production of the specific types of cheese. In 1996, those strains that had been identified as suitable on the basis of their acidifying activity were further assessed using bacteriophage strains from CSL’s collection and suitable bacterial strains were added to their starter culture collection . This research enabled CSL to increase the number of strains used in rotation in starter cultures for cheese making. In addition, research on the composition of the natural starter for Fiordilatte enabled CSL to select a new starter culture containing mesophilic cocci and S. thermophilus strains.
IZCS, Italy have given their microbial strains to small dairy factories in Sardinia for limited production of artisanal cheeses at an industrial level.
One of the strains isolated by CSIC, Spain and another isolated by Teagasc, Ireland produced potentially useful bacteriocins. Teagasc continued research in-house to develop a phage-resistant, bacteriocin-producing plasmid that underwent commercial trials at the end of 1998. One patent has been granted and two others have been applied for relating to the use of the bacteriocin from L. lactis DPC3147 in different applications. In early 1999, a license was granted to the Cross Vetpharm Group Ltd, Ireland to use the bacteriocin in developing a treatment for mastitis. The isolate also has applications in controlling growth of non-starter lactic acid bacteria in cheese and in inhibiting Listeria on the surface of cheese. Teagasc are currently discussing license agreements with three other commercial companies.
Manufacture of cheddar cheese and its territorial varieties require rapid acid-producing mesophilic starters. Unfortunately, very few of the mesophilic isolates (mainly researched at Teagasc) in the project were rapid acid-producers. Those that were gave atypical cheese flavours. Thermophilic strains were potentially more useful.
A lactic acid bacteria collection (ACA-DC), accepted at the World Federation Culture Collection, has been created at the Agricultural University of Athens, Greece. This contains more than 300 strains isolated from traditional Greek cheese and yogurt. A new species, Streptococcus macedonicus, was identified from this collection. National research attempted to fund commercialization of these cultures, however this failed due to the cost of commercial production. Thus, they require finance to procure a modern system to pack cultures and start commercialization. They are also preparing a new collection of yeasts and lactic acid bacteria from traditional sour dough for the preparation of bread.
No further research has been carried out on the 700 plus strains isolated and characterized at INIA, Madrid.
Nizo considered this to be a very successful project. They have carried out (confidential) follow-up contract research projects in this field. The clearly believe that there is strong interest from the dairy industry, as well as from starter-producing companies and the ingredient industry. Some work has been continued by PhD students and Nizo are currently trying to initiate a new project with strong contribution from industry; 4 companies have already indicated they wish to participate. This project would include ECLAIR partners and would be coordinated by the original participant from UCSC, Italy. They are confident that this approach will lead to several applications (new dairy products) although considerable basic and applied research still has to be done.
Impact
Commercial
As a result of the further in-house research, CSL (Italy) has added a new starter (STAR-IDC) to its product range. STAR-IDC contains S. thermophilus and mesophilic flavouring bacteria and is suitable for production of pasta filata cheeses, mozzarella cheese, etc.
Patent No 5950269 (1995) has been granted for the bacteriocin from L. lactis DPC3147. There is significant commercial interest in the results and subsequent research from this project. However, it may be a number of years before these results are fully commercialized. The development work carried out by Cross VetPharm Group Ltd is very promising with application in prevention of mastitis in dairy cows. They anticipate that they will have a product with a Europe-wide market ready for registration in 2000.
In the Netherlands, there is strong interest from the dairy industry, as well as from starter-producing companies and the ingredient industry.
IZCS indicated that the intensive studies of the microbial composition of the most important traditional and protected designation of origin (PDO) cheeses from Sardinia carried out in this project were invaluable. IZCS are a research institute and thus cannot sell starter cultures, however the microbial isolates collected and stored at their laboratory are available to industry on request. These have often been given to small dairy factories that were attempting to produce artisanal cheeses at an industrial level. Production has been limited to the local market thus information on commercialization is not available.
Associated
A good network of research and commercial contacts has been established. IZCS has carried out further research supported by COST action ’95 and regional and national funds. Research continues, with a new project being put together to further exploit the results of this project.
Further information
Bianchi Salvadori, B., Guerriero, P. (1995) Characteristics of Streptococcus thermophilus strains isolated from some Italian natural starters, curds and cheeses. Poster presented at the European Commission Biotechnology and FAIR Programmes Conference "Lactic Acid Bacteria – From Fundamental Research to Innovative Applications" (Cork, Ireland, 22-26 October 1995)
Bianchi Salvadori, B. (1995) Lactic acid bacteria and acidification. Report presented at the European Symposium "Contribution of Fermentations to the Agro-Food Sector" (Dole, France, 19-21 September 1995, sponsored by the EU and organised by the Pole Européen de Communication et d’Echanges Agro Alimentales (PEA))
Bianchi Salvadori, B., Rottigni, C. (1995) Impiego degli starter lattici per la fabbricazione dei prodotti tipici sardi. Report presented at the 25th National Congress of the Società Italiana di Microbiologia (Alghero, Porte Conte, Sardinia, 27-30 September 1995). In: Ind. Latte XXXI, 4: 25-37
Bianchi Salvadori, B., Guerriero, P., Palladino, L., Lupo, R. (1996) Selezione di ceppi di Streptococcus thermophilus per la formulazione di starter specifico per il formaggio Caciotta. Ind. Latte XXXII, 4: 21-48
Bianchi Salvadori, B., Guerriero, P., Lavezzari, D., Minissi, S. (1996) Industrial reproduction of Italian Mozzarella cheese by means of strains isolated from their own natural starter. Poster presented at the IDF Symposium "Ripening and Quality of Cheeses" (Besançon, France, 26-28 February 1996). In: IDF Bulletin, 317: 53
Cogan, T.M. and Rea, M.C. (1996) Artisanal European Cheeses. EUR 16788 EN, OOPEC, Luxembourg.
Cogan, T.M., Barbosa, M., Beuvier, E., Bianchi Salvadori, B., et al. (1997) Characterization of the lactic acid bacteria in artisanal dairy products. J. Dairy Res., 64: 409-421
Dougherty, B., Hill, C., Weidmann, J.F., Richardson, D.R., Venter, J.C. and Ross, R.P. (1998) Sequence and analysis of the 60 kb conjugative, bacteriocin producing plasmid pMRCo1 from Lactococcus lactis DPC3147. Mol. Microbiol. 29: 1029-1038
McAuliffe, O., Ryan, M.P., Ross, R.P., Hill, C., Breeuwer, P. and Abee, T. (1998). Lacticin 3147 a broad-spectrum bacteriocin which selectively dissipates the membrane potential. Appl. Environ. Microbiol. 64: 439-445
Ryan, M.P., Meaney, W.J., Ross, R.P. and Hill, C. (1998) Evaluation of Lacticin 3147 and a teat seal containing this bacteriocin for inhibition of mastitis pathogens. Appl. Environ. Microbiol. 64: 2287-2290
Ryan, M.P., Rea, M.C., Hill C. and Ross, R.P. (1996) An application of cheddar cheese manufacture for a strain of Lactococcus lactis producing a novel broad-spectrum bacteriocin, lacticin 3147. Appl. Environ. Microbiol. 62: 612
Tsakalidou, E., Zoidou, E., Pot, B., Wassill, L., Ludwig, W., Devriese, L.A., Kalantzopoulos, G., Schleifer, K.H., Kesters, K. (1998) Identification of streptococci from Greek kasseri cheese and description of Streptococcus sp. nov. Int. J. Systematic Bacteriology 48: 519-527
Contacts
Author
CSL
Cross VetPharm Group Ltd
INIA
IZCS
Nizo Food Research
TEAGASC
UCSC
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by CPL Press:
03/07/2007
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