PROLAB 77 / 2015

Rezultate etapa IV/2015

În ultimii ani studiile realizate privind obţinerea de produse de tip probiotic şi elucidarea mecanismelor implicate în îmbunătăţirea stării de sănătate a consumatorului, au evidenţiat nu numai complexitatea interacţiunilor dintre microorganismele probiotice şi organismul gazdă (inclusiv microbiota de la nivelul tractului gastrointestinal), dar şi faptul că efectele sunt tulpină specifice. Pentru obţinerea unui efect probiotic cumulat şi potenţat în cadrul prezentului proiect obiectivul major este reprezentat de realizarea de consorţii de bacterii lactice cu efect probiotic.
Obiectivele operaţionale ale etapei IV-2015 au urmărit: caracterizarea compuşilor cu acţiune antimicrobiană produşi de bacterii lactice şi influenţa condiţiilor de creştere; influenţa consorţiilor asupra microbiotei umane simulate în sistemul GIS2; analiza din punct de vedere chimic a probelor prelevate din sistemele GIS2; analiza cantitativă a influenţei consorţiilor de bacterii lactice asupra profilului compuşilor secretaţi de celulele epiteliale şi evaluarea citotoxicitatii consorțiilor şi identificarea taxonomică a tulpinilor de bacterii lactice prin tehnici de biologie moleculară. 
Toate obiectivele propuse în cadrul prezentei etape au fost îndeplinite şi din cele 10 consorţii realizate în cadrul etapei anterioare, pe baza studiilor desfăşurate au fost selectate şi analizate 3 care au prezentat activitate antimicrobiană şi/sau activitate imunomodulatrie potenţată.
O parte din rezultate au fost diseminate prin publicare în 4 articole ISI (1 articol publicat şi 3 acceptate), 2 articole BDI, un brevet şi 4 participãri la manifestãri ştiinţifice internationale.
Este de menţionat faptul că sistemul GIS2 a fost premiat cu medalia de aur la salonul EuroInvent 2015.

Publicatii stiintifice:
Articole in reviste ISI
  1. Grosu-Tudor S.S., Zamfir M., Van der Meulen R., De Vuyst L., 2013, Isolation of novel homopolysaccharide-producing lactic acid bacteria from Romanian raw milk and fermented dairy products, European Food Research and Technology 237:609-615.
  2. Vamanu E., Pelinescu D.R., Avram I., Nita S., 2013, An in-vitro evaluation of antioxidant and colonic microbial profile levels following mushroom consumption, BioMed Research International http://dx.doi.org/10.1155/2013/289821
  3. Vamanu E., 2014, Testing in vitro viability of thermophilic probiotic strain in simulated gastrointestinal conditions,  Annals of Microbiology 64(3):1439-1442
  4. Grosu-Tudor S.-S., Stancu M.-M., Pelinescu D.,  Zamfir M., 2014, Characterization of some bacteriocins produced by lactic acid bacteria isolated from fermented foods, World Journal of Microbiology and Biotechnology 30: 2459–2469.
  5. Vamanu E., Sarbu I., Nedelcu I., Pelinescu D., 2015, Study of PROEXO product influence on infant microbiota in an in vitro colonic fermentation system, Annals of Microbiology 65(2): 1189-1193.
  6. Ditu L. M., Chifiriuc M. C., Pelinescu D., Sarbu I., Pircalabioru G., Mihaescu G., 2014, Class I and II Bacteriocins: Structure, Biosynthesis and Drug Delivery Systems for the Improvement of their Antimicrobial Activity, Current Proteomics  11: 121-127.
  7. Gatea F., Teodor E., Paun G., Matei A., Radu G. L., 2015, Capillary Electrophoresis Method Validation for Organic Acids Assessment in Probiotics, Food Analytical Method 8 (5): 1335-1340.
  8. Moroeanu V. I., Vamanu E., Paun G., Neagu E., Ungureanu O. R., Eremia S. A., Radu G. L, Ionescu R.,  Pelinescu D. R, 2015,  Probiotic strains influence on infant microbiota in the in vitro colonic fermentation model GIS1, Indian Journal of Microbiology, 55(4): 423-429
  9. Vamanu E, 2015,  Effect of gastric and small intestinal digestion on lactic acid bacteria activity in a GIS1 simulator, Saudi Journal of Biological Sciences– Elsevier, on-line paper
  10. Sarbu I., Vassu T., Stoica I., Marutescu L., Chifiriuc C.,  Pelinescu D., 2015,  Phenotypic and genotypic assessment of Lactobacillus plantarum influence on Candida albicans fluconazole resistance,  Annals of Microbiology, 1-7.
  11. Zamfir M., Stefan I., Grosu-Tudor S. S., 2015,  Influence of growth medium composition on the bacteriocin activity of some lactic acid bacteria, Romanian Biotechnological Letters, in press

Articole in reviste BDI
  1. Ditu L.M., Chifiriuc M. C.,  Marutescu L.,  Bleotu C.,  Pelinescu D.,  Bezirtzoglou E., Mihaescu G., Lazar V., 2014, Immunomodulatory effect of non-viable components of probiotic culture stimulated with heat-inactivated Escherichia coli and Bacillus cereus on holoxenic mice, Microbial Ecology In Health & Disease 25, p.1-6.
  2. Ditu L. M., Chifiriuc M.C., Pelinescu D., Sarbu I., Lazar V., Mihaescu G., 2014, Two-Component Regulatory Systems – implication in the quorum sensing mechanisms and bacteriocin production in lactic acid bacteria, Biointerface Research in Applied Chemistry  4(4), p. 813-815.
  3. Zamfir M., Cornea P.C., De Vuyst L., Grosu-Tudor S.-S., 2014, Biodiversity and biotechnological potential of lactic acid bacteria, AgroLife Scientific Journal 3(1), p. 169-176.
  4. Vamanu E., 2015, Biochemical properties and viability of two thermophilic lactic acid bacteria in a single chamber gastrointestinal tract simulator –GIS1: partial characterizations, University of Agricultural Sciences and Veterinary Medicine Iasi, Lucrări Ştiinţifice-Seria Zootehnie, 312 – 315.
  5. Sarbu I., Vassu T., Stoica I., Chifiriuc C. Bucur M., Rusu E., Pelinescu D., 2015, Analysis on the antimicrobial activity of some lactic acid bacteria strains, Journal of Infectious Diseases 18(2-3): 87-91.

Prezentari la conferinte 
  1. Grosu – Tudor S.S., Zamfir M., 2012,  Functional properties of lactic acid bacteria isolated from Romanian fermented vegetables, II International Conference on Antimicrobial Research, Lisabona, Portugalia, 21-23 noiembrie 2012, p. 563.
  2. Grosu-Tudor S.-S., Stancu M.-M., Pelinescu D.,  Zamfir M., 2014, Characterization of some bacteriocins produced by lactic acid bacteria isolated from fermented foods, 11th Symposium on Lactic Acid Bacteria, Egmond Aan Zee/Olanda, 31august-04 septembrie
  3. Sarbu I., Vassu T., Stoica  I., Marutescu L., Chifiriuc C., Nedelcu I., Pelinescu D., 2014, The influence of lactic acid bacteria strains on the antifungal resistance mechanisms of Candida albicans strains, 11th Symposium on Lactic Acid Bacteria, Egmond Aan Zee/Olanda, 31august-04 septembrie
  4. Sarbu I.,  Tanase A.M., Zamfir M.,  Vassu T., Stoica I., Chifiriuc C.M., Ionescu R., Pelinescu D., 2014, Screening for antimicrobial activity of lactic acid bacteria strains isolated from different Romanian food products, 3rd International Conference on Clinical Microbiology and Microbial Genomics, Valencia/Spania, 24-26 septembrie
  5. Sarbu I., Ionescu R. Vassu T., Stoica I., Ditu L.M., Marutescu L., Chifiriuc C.M., Petrut S., Pelinescu D., 2014, Study of some lactic acid bacteria strains probiotic effect  for use in gastrointestinal infections, 3rd International Conference on Clinical Microbiology and Microbial Genomics, Valencia/Spania, 24-26 septembrie
  6. Chifiriuc M.C., Marutescu L., Ionescu B., Ionescu D., Gheorghe I., Czobor I., Mihaescu G., Lazar V., 2014, Flow cytometry characterisation of gram positive and gram negative bacterial strains exhibiting non-enzymatic and enzymatic resistance mechanisms, 14TH EUROCONFERENCE ON CLINICAL CELL ANALYSIS, Lisabona, Portugalia, 13 -18 septembrie 2014.
  7. Zamfir M., Cornea P.C., De Vuyst L., Grosu-Tudor S.-S., 2014, Biodiversity and biotechnological potential of lactic acid bacteria, International Conference "Agriculture for Life, Life for Agriculture, Bucuresti/Romania, 5-7 iunie – prezentare orala
  8. Zamfir M., Grosu-Tudor S.S., Stancu M.M., Cornea C.P., De Vuyst L., 2015, Biodiversity of lactic acid bacteria from traditional Romanian fermented foods. Conferință invitată la European Biotechnology Congress, București, 7-9 mai
  9. Zamfir  M., Stancu  M.M., Stefan  I.R., Grosu-Tudor S.S., 2015, Bacteriocins produced by lactic acid bacteria isolated from Romanian traditional fermented foods. BioMicroWorld 2015, Barcelona, 28-30 octombrie 2015- Prezentare orală
  10. Biochemical properties and viability of two thermophilic lactic acid bacteria in a single chamber gastrointestinal tract simulator- GSI1: partial characterizations, 2015, Lucrări Științifice- Seria Zootehnie- University of Agricultural Sciences and Veterinary Medicine, Iași
Premii:
Partener USAMVB - autor Emanuel Vamanu
  1. Medalie de aur - Salonul Internaţional de Invenţii de la Geneva (ELVEŢIA 2013)  GIS1 – Sistem unicameral de simulare in vitro a tractului gastrointestinal uman
  2.  Medalie de aur –Expoziția Europeană a Creativității și Inovării (Iasi 2015). GIS2 – sistem in vitro de simulare a colonului uman în trei segmente










Results phase IV/2015

In last decades studies regarding probiotic products for improving the health of the consumer, were focused on obtaining new probiotic strains and highlighted the complexity of interactions between probiotic microorganisms and the human host (including microbiota of the gastrointestinal tract), finding that  positive effects were usually strain specific. The major goal of this project was to obtain strains and consortia of lactic acid bacteria with enhanced probiotic effect.
The operational objectives of phase IV-2015 were focused on: characterization of antimicrobial compounds produced by lactic acid bacteria and influence on culture conditions; the influence of consortia on human microbiota simulated in GIS2 system; chemically analysis of samples taken from GIS2 system; quantitative analysis of the compounds released by the epithelial cells in presence of lactic acid bacteria consortia, the cytotoxicity assay and the taxonomic identification of lactic acid bacteria strains by molecular techniques.
All objectives proposed in this phase have been achieved and 3 consortia were selected out of the 10 consortia obtained in the previous stage. The 3 selected consortia presented antimicrobial activity and enhanced immunomodulatory effect.
Some of the results were disseminated through publication in 4 ISI papers (3 of them were accepted and one was published), 2 BDI papers, 1 patent and 4 participation in international conferences. It is it worth mentioning that the GIS2 system was awarded with the gold medal at the salon EUROINVENT 2015.

Scientific publications:
  ISI journals
  1.  Grosu-Tudor S.S., Zamfir M., Van der Meulen R., De Vuyst L., 2013, Isolation of novel homopolysaccharide-producing lactic acid bacteria from Romanian raw milk and fermented dairy products, European Food Research and Technology 237:609-615.
  2. Vamanu E., Pelinescu D.R., Avram I., Nita S., 2013, An in-vitro evaluation of antioxidant and colonic microbial profile levels following mushroom consumption, BioMed Research International http://dx.doi.org/10.1155/2013/289821
  3. Vamanu E., 2014, Testing in vitro viability of thermophilic probiotic strain in simulated gastrointestinal conditions,  Annals of Microbiology 64(3):1439-1442
  4. Grosu-Tudor S.-S., Stancu M.-M., Pelinescu D.,  Zamfir M., 2014, Characterization of some bacteriocins produced by lactic acid bacteria isolated from fermented foods, World Journal of Microbiology and Biotechnology 30: 2459–2469.
  5. Vamanu E., Sarbu I., Nedelcu I., Pelinescu D., 2015, Study of PROEXO product influence on infant microbiota in an in vitro colonic fermentation system, Annals of Microbiology 65(2): 1189-1193.
  6. Ditu L. M., Chifiriuc M. C., Pelinescu D., Sarbu I., Pircalabioru G., Mihaescu G., 2014, Class I and II Bacteriocins: Structure, Biosynthesis and Drug Delivery Systems for the Improvement of their Antimicrobial Activity, Current Proteomics  11: 121-127.
  7. Gatea F., Teodor E., Paun G., Matei A., Radu G. L., 2015, Capillary Electrophoresis Method Validation for Organic Acids Assessment in Probiotics, Food Analytical Method 8 (5): 1335-1340.
  8. Moroeanu V. I., Vamanu E., Paun G., Neagu E., Ungureanu O. R., Eremia S. A., Radu G. L, Ionescu R.,  Pelinescu D. R, 2015,  Probiotic strains influence on infant microbiota in the in vitro colonic fermentation model GIS1, Indian Journal of Microbiology, 55(4): 423-429
  9. Vamanu E, 2015,  Effect of gastric and small intestinal digestion on lactic acid bacteria activity in a GIS1 simulator, Saudi Journal of Biological Sciences– Elsevier, on-line paper
  10. Sarbu I., Vassu T., Stoica I., Marutescu L., Chifiriuc C.,  Pelinescu D., 2015,  Phenotypic and genotypic assessment of Lactobacillus plantarum influence on Candida albicans fluconazole resistance,  Annals of Microbiology, 1-7.
  11. Zamfir M., Stefan I., Grosu-Tudor S. S., 2015,  Influence of growth medium composition on the bacteriocin activity of some lactic acid bacteria, Romanian Biotechnological Letters, in press
 IDB journals
  1. Ditu L.M., Chifiriuc M. C.,  Marutescu L.,  Bleotu C.,  Pelinescu D.,  Bezirtzoglou E., Mihaescu G., Lazar V., 2014, Immunomodulatory effect of non-viable components of probiotic culture stimulated with heat-inactivated Escherichia coli and Bacillus cereus on holoxenic mice, Microbial Ecology In Health & Disease 25, p.1-6.
  2. Ditu L. M., Chifiriuc M.C., Pelinescu D., Sarbu I., Lazar V., Mihaescu G., 2014, Two-Component Regulatory Systems – implication in the quorum sensing mechanisms and bacteriocin production in lactic acid bacteria, Biointerface Research in Applied Chemistry  4(4), p. 813-815.
  3. Zamfir M., Cornea P.C., De Vuyst L., Grosu-Tudor S.-S., 2014, Biodiversity and biotechnological potential of lactic acid bacteria, AgroLife Scientific Journal 3(1), p. 169-176.
  4. Vamanu E., 2015, Biochemical properties and viability of two thermophilic lactic acid bacteria in a single chamber gastrointestinal tract simulator –GIS1: partial characterizations, University of Agricultural Sciences and Veterinary Medicine Iasi, Lucrări Ştiinţifice-Seria Zootehnie, 312 – 315.
  5. Sarbu I., Vassu T., Stoica I., Chifiriuc C. Bucur M., Rusu E., Pelinescu D., 2015, Analysis on the antimicrobial activity of some lactic acid bacteria strains, Journal of Infectious Diseases 18(2-3): 87-91.
  Congress presentations
  1. Grosu – Tudor S.S., Zamfir M., 2012, “Functional properties of lactic acid bacteria isolated from Romanian fermented vegetables”, II International Conference on Antimicrobial Research, Lisabona, Portugalia, 21-23 noiembrie 2012, p. 563.
  2. Grosu-Tudor S.-S., Stancu M.-M., Pelinescu D.,  Zamfir M., 2014, Characterization of some bacteriocins produced by lactic acid bacteria isolated from fermented foods, 11th Symposium on Lactic Acid Bacteria, Egmond Aan Zee/Olanda, 31august-04 septembrie
  3. Sarbu I., Vassu T., Stoica  I., Marutescu L., Chifiriuc C., Nedelcu I., Pelinescu D., 2014, The influence of lactic acid bacteria strains on the antifungal resistance mechanisms of Candida albicans strains, 11th Symposium on Lactic Acid Bacteria, Egmond Aan Zee/Olanda, 31august-04 septembrie
  4. Sarbu I.,  Tanase A.M., Zamfir M.,  Vassu T., Stoica I., Chifiriuc C.M., Ionescu R., Pelinescu D., 2014, Screening for antimicrobial activity of lactic acid bacteria strains isolated from different Romanian food products, 3rd International Conference on Clinical Microbiology and Microbial Genomics, Valencia/Spania, 24-26 septembrie
  5. Sarbu I., Ionescu R. Vassu T., Stoica I., Ditu L.M., Marutescu L., Chifiriuc C.M., Petrut S., Pelinescu D., 2014, Study of some lactic acid bacteria strains probiotic effect  for use in gastrointestinal infections, 3rd International Conference on Clinical Microbiology and Microbial Genomics, Valencia/Spania, 24-26 septembrie
  6. Chifiriuc M.C., Marutescu L., Ionescu B., Ionescu D., Gheorghe I., Czobor I., Mihaescu G., Lazar V., 2014, Flow cytometry characterisation of gram positive and gram negative bacterial strains exhibiting non-enzymatic and enzymatic resistance mechanisms, 14TH EUROCONFERENCE ON CLINICAL CELL ANALYSIS, Lisabona, Portugalia, 13 -18 septembrie 2014.
  7. Zamfir M., Cornea P.C., De Vuyst L., Grosu-Tudor S.-S., 2014, Biodiversity and biotechnological potential of lactic acid bacteria, International Conference "Agriculture for Life, Life for Agriculture, Bucuresti/Romania, 5-7 iunie – prezentare orala
  8. Zamfir M., Grosu-Tudor S.S., Stancu M.M., Cornea C.P., De Vuyst L., 2015, Biodiversity of lactic acid bacteria from traditional Romanian fermented foods. Conferință invitată la European Biotechnology Congress, București, 7-9 mai
  9. Zamfir  M., Stancu  M.M., Stefan  I.R., Grosu-Tudor S.S., 2015, Bacteriocins produced by lactic acid bacteria isolated from Romanian traditional fermented foods. BioMicroWorld 2015, Barcelona, 28-30 octombrie 2015- Prezentare orală
  10. Biochemical properties and viability of two thermophilic lactic acid bacteria in a single chamber gastrointestinal tract simulator- GSI1: partial characterizations, 2015, Lucrări Științifice- Seria Zootehnie- University of Agricultural Sciences and Veterinary Medicine, Iași

 AWARDs:
Partner USAMVB - author Emanuel Vamanu
  1. Gold medal - Exhibition of Inventions  Geneva (Switzerland 2013)  GIS1 system – Single chamber system for in vitro simulation of the human gastrointestinal tract
  2. Gold medal -  European Exhibition of Creativity and Inobationn (Iasi 2015) – GIS2 – In vitro simulation system of human colon in three stages