CEEX 9/2005, PROBIO, english version

Title: Biotechnological and molecular studies on probiotic products for farm animals. Acronym: PROBIO

Project director: Prof. Dr. Tatiana Vassu-Dimov

Main objectiv of the project was to investigate the obtaining of probiotic products by biotechnological and molecular techniques.

Abstract:

This project had a high degree of complexity and implied the participation of seven partners during three years. The main products resulted from experimental studies included in 3 categories, namely: original research papers, patents and the lab-scale production of the probiotic product.
In terms of scientific production, the researchers involved in this project have already developed 11 original scientific papers that included the most significant scientific results obtained from: isolation; phenotypic and molecular characterization of microbial strains; probiotics capacity proved by in vitro and in vivo models and biotechnology studies (optimization of growth conditions).
Experimental results obtained during the performance of this project enabled development of two patents submitted at OSIM having the following registration numbers: A00655 and A00504/17.07.2007 / 18.09.2007.
The project led to obtaining a probiotic product consisting of 3 strains of yeast (Saccharomyces cerevisiae 1-29, Saccharomyces cerevisiae R-BF, Kluyveromyces marxianus R-CS) and two strains of lactic acid bacteria (Lactobacillus paracasei CMGB16, Enterococcus faecium CMGB 8). Microorganisms were grown at micro scale in economical culture media and growth parameters (aeration, pH values, etc) were optimized in order to obtain a high biomass production. Selected microorganisms were mixed in certain proportions, and the product obtained was stored at low temperature (4ºC). Following tests carried out in vitro and in vivo proved the beneficial effect of probiotic product in terms of: increasing the immune response, ability to inhibit pathogenic microorganisms (both by adhering to host cells and through the synthesis of compounds with antimicrobial action such as organic acids, bacteriocins and killer toxins), and in terms of decreasing concentration of cholesterol. Overall, we found that the product developed by us lead to a general improvement of health of animals. Also, it is important to note this product can be a successful alternative in the treatment of infections with common pathogenic strains of Escherichia coli and Salmonella, thereby reducing the risk of antibioresistance.


Partners:

Coordinator: Center for Research, Training and Consulting in Microbiology, Genetics & Biotechnology – Faculty of Biology, University of Bucharest – MICROGEN

Partner 1: University of Agronomic Science and Veterinary Medicine Bucharest – USAMVB

Partner 2: Institute of Biology Bucharest - IBB

Partner 3: National Institute of Research and Development for Microbiology and Immunology “Cantacuzino”–IC

Partner 4: Pasteur Institute - IP

Partner 5: DEXTER COM S.R.L.

Partner 6: National Institute of Development – Research in Biological Science Bucharest – INCDSB

DIAGRAM OF PROJECT EXECUTION:

Task 1: Elaborating the set of techniques, isolation and selection of some microorganisms with probiotic effect (MICROGEN, IBB, USAMVB)

Task 2: Genetical, biochemical and physiological characterisation of selected lactic acid bacteria and elaboration of methods for bacterial biomass obtaining (IBB, MICROGEN, DEXTER, IP, USAMVB)

Task 3: Genetical, biochemical and physiological characterisation of the selected yeast strains and elaboration of methods for yeast biomass obtaining (MICROGEN, IC, USAMVB)

Task 4: Studies concerning the mechanisms involved in the probiotic effect of the selected microorganisms (DEXTER, INCDSB, IBB, MICROGEN, IP, IC)

Task 5: Elaboration and characterisation of the probiotic compound (USAMVB, MICROGEN, INCDSB, IC)

Task 6: Final evaluations of the characteristics and stability of the probiotic compound (MICROGEN, USAMVB, IP)

Publications

Books

1. Vamanu A., Popa O., Vamanu E., Sisteme productive în biotehnologii / Ed. ARS Docendi, ISBN 973-558-232-5

2. Vamanu A., Vamanu E., Sisteme productive în biotehnologii - Lucrari practice/ Ed. ARS Docendi, ISBN 978-973-558-322-4

Scientific papers in international journals

1. M. Vancanneyt, M. Zamfir, M. De Wachter, I. Cleenwerck, B. Hoste, F. Rossi, F. Dellaglio, L. De Vuyst, J. Swings, 2006. Reclassification of Leuconostoc argentinum as a later synonym of Leuconostoc lactis, International Journal of Systematic and Evolutionary Microbiology, 56: 213-216.

2. M. Zamfir, M. Vancanneyt, L. Makras, F. Vaningelgem, K. Lefebvre, B. Pot, J. Swings, L. De Vuyst, 2006. Biodiversity of lactic acid bacteria in Romanian dairy products, Systematic and Applied Microbiology, 29: 487-495.

Scientific papers in national journals

1. Vamanu E., Vamanu A., Popa O., Pelinescu D., Avram I., Enache Soare S., 2007, Obtaining probiotic biomass from yeasts used for fodder farm animals, Archiva Zootechnica, vol. 10 p. 35 – 45.

2. Popa V., Pirvulescu M., Samarineanu M., Pelinescu D., Vassu- Dimov T., Stoica I., Sasarman E., Caplan E., Botus D., Nica I., Alexandra Alecu 2008, Studies regarding to the effect of some Enterococcus faecium strains in avian infections with Salmonella gallinarum, Listeria monocytogenes and Escherichia coli, Lucrări Stiinţifice de Medicină Veterinară (Scientifical Papers Veterinary Medicine, CNCSIS B259) USAMV Timişoara, vol. XXXXI, ISSN:1221-5295 (in press)

3. Popa V., Pirvulescu M., Samarineanu M., Pelinescu D., Vassu – Dimov T., Stoica I., Sasarman E., Caplan E., Botus D., Nica I., Alecu A., 2008, Efectul unor tulpini de Enterococcus faecium in infectiile aviare cu Salmonella gallinarum, Listeria monocytogenes si Escherichia coli, Magazinul Avicol / Buletin informativ – Societatea Medicilor Veterinari in Patologie aviara si a Animalelor Mici din Romania 3 (18) / 19 (74-2): 28-34.

4. Teodor E., Bîrsan C., Smarandache D., Penu R., Radu G. L., Determination of hydrosoluble vitamins content and antioxidant activity of some yeast strains with probiotic potential, Revista de Chimie, 2007, 58, 12, 1249-1252.

5. Barsan C., Teodor E., Vassu T., Radu G. L., Determination of organic acids content by liquid chromatography (HPLC) from probiotic potential extracts, Rom. Biol. Sci., 2007, vol. V, nr. 1-4, p. 13-20.

6. Vamanu E., Vamanu A., Popa O., Pelinescu D., Avram I., Enache Soare S., 2007, The resistance determination of two lactic bacteria strains for the obtaining of probiotic biomass, Annals of West University of Timisoara, Series of Chemistry, 16, 2, 108.

7. Popa V., Caplan E., Pirvulescu M., Botuş D., Mihăilescu R., Samarineanu M., Popovici A., Vassu T., Smarandache D., Enterococcus faecium şi Lactobacillus paracasei – potential probiotics strains, Buletin informativ, 2007, 70(2): 18, p.9-13.

Scientific papers presented as oral or posters at national/international congresses:

  1. Albu C., Teodor E., Determinarea acizilor organici prin HPLC din preparate cu potential probiotic, Work-shop - Bioanaliza versus Biotehnologia, ed. A IV-a, Bucuresti 14 dec. 2007.
  2. Boghean R., Tucureanu C., Pelinescu D., Lerescu L., Vassu-Dimov T., Salageanu A., „Immunological properties of probiotics: protection against experiemntal bacterial infection and induction of cytokine secretion”, 5th Balkan Congress of Immunology, June 01-03, Ohrid, Republic of Macedonia, Maced J Med Sci 2008, 1 (Suppl. 1):S49, ISSN 1857-5449
  3. Bucur E. O., Valentina Diaconu I., Caplan M. E., Popa V., Pirvulescu M., Pelinescu D., Aspecte microscopice ale organelor participante la răspunsul imun la păsări inoculate cu preparate probiotice, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  4. Chifiriuc C. M., Sasarman E., Lazar V., Smarandache D., Vassu T., Stoica I., Dracea O., Iordache C., Bucur M., Larion C., Avram O., Studii preliminare de selectare a unui nou produs probiotic cu utilizare în profilaxia şi tratamentul bolii diareice la copilul mic, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  5. Grosu-Tudor S., Zamfir M., Prebiotice, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  6. Lazar V., Balotescu M.C., Ditu L.M., Bleotu C., Smarandache D., Soare S., Larion C., Cotar A., Cernat R.. In vitro study of the influence of cell-free liquid lactobacillus cultures on the pathogenic features of staphylococcus aureus clinical strains. 2nd Fems Microbiology Congress, Madrid, Spain, 4-8 July 2006.
  7. Pelinescu D., Lazar V., Balotescu C., Stoica I., Sasarman E., Ghindea R., Avram I., Nohit A.-M., Ionescu R., Soare S., Csutak O., Vassu T., Caracterizarea genetica, biochimica si fiziologica a microorganismelor cu potenţial probiotic, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  8. Pelinescu D., Stoica I., Balotescu C., Sasarman E., Bleotu C., Avram I., Soare-Enache S., Ionescu R., Vamanu E., Serbancea F., Vassu T., 2008, Lactic acid bacteria characterization and in vitro studies on probiotic effect, Abstracts Book of Genomes 2008 – Functional genomics of microorganisms, Paris Apr. 8-11, p.178 (P113)
  9. Pelinescu D., Stoica I., Sasarman E., Balotescu C., Nohit A., Ionescu R., Csutak O., Vassu Dimov T., 2006, Identification and characterization of some lactic acid bacteria strains isolated from fermented plants, Abstracts Book of The Second FEMS Congress of European Microbiologists, Spain, Madrid, July 4-8, 2006, p 203 (P.FOD.74).
  10. R. Boghean, C. Tucureanu, L. Lerescu, D. Pelinescu, T. Vassu-Dimov, A. Salageanu, “Proprietati imunologice ale bacteriilor lactice: modele in vivo si in vitro”, Sesiunea Stiintifica Anuala a I.N.C.D.M.I. “Cantacuzino”, ianuarie 2008
  11. Radu G. L., Teodor E., Albu C., Determinarea conţinutului de vitamine hidrosolubile şi de acizi organici prin cromatografie de lichide (HPLC) din extracte cu potenţial probiotic, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  12. Sasarman E., Vassu T., Pelinescu D., Probioticele, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  13. Savu L., Pelinescu D., Studii in vitro pe tulpinile de bacterii lactice selecţionate pentru determinarea compuşilor cu acţiune antimicrobiana si caracterizarea acestora, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  14. Tucureanu C., Diana Pelinescu, Lucian Lerescu, Tatiana Vassu Dimov, Raluca Boghean si Aurora Salageanu, „Studiul proprietatilor imunologice ale probioticelor: protectia in infectii bacteriene experimentale si inducerea secretiei de citokine”, A XXXVII -a Conferinta Nationala de Imunologie, Iasi, 19-21 Septembrie 2007, ISBN 978-973-716-707-1
  15. Tucureanu C., Pelinescu D., Lerescu L., Vassu Dimov T., Boghean R., Salageanu A., Studiul proprietăţilor imunologice ale probioticelor: protecţia in infecţii bacteriene experimentale si inducerea secreţiei de citokine , Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  16. Vamanu A., Vamanu E., Pelinescu D., Popa O., Produs probiotic pentru creşterea performanţelor zootehnice la animalele de fermă, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  17. Vamanu E., Vamanu A., Popa O., Pelinescu D., Avram I., Enache Soare S., Obtaining probiotic biomass from yeasts used for fodder farm animals, 2007, Proceedings of the 6th IBNA International symposium, 27 – 28 septembrie, 2007, Balotesti.
  18. Vassu T., Pelinescu D., Stoica I., Sasarman E., Cercetări biotehnologice şi de nivel molecular privind obţinerea de preparate probiotice de uz zoo-veterinar, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  19. Zamfir M., Grosu-Tudor S., Popescu G., Carasan M.-E., Cercetări privind cinetica sintezei de bacteriocine de către bacterii lactice izolate din produse lactate fermentate tradiţional, Workshop – PROBIOTICELE. Alternativă la tratamentele convenţionale, Universitatea din Bucureşti – Facultatea de Biologie, 20 iunie 2008.
  20. Zamfir M., Grosu-Tudor S.S., 2007, Studii la nivel molecular asupra unor bacterii lactice cu aplicaţii biotehnologice, Sesiunea Anuala de Comunicari Stiintifice a Institutului de Biologie Bucuresti

Patents

1. Brevet de inventie nr. A00504/17.07.2007: Enterococcus faecium CMGB 8, bacterie lactică de tip ptobiotic pentru uz zootehnic,

Autori: D. Pelinescu, C. M. Chifiriuc, E. Săsărman, T. Vassu Dimov, I. Stoica, S. Enache Soare, R. Ionescu, I. Avram,

  1. Brevet de inventie nr. A00655/ 18.09.2007 : Izolarea, selecţia şi caracterizarea unei tulpini de Lactobacillus paracasei ssp. paracasei de tip probiotic pentru uz uman

Autori: Sasarman E., Chifiriuc C.M., Pelinescu D., Bleotu C., Vassu T., Stoica I.