Development of nutraceutical products with probiotics

U ovom radu su razvijeni novi nutraceutički proizvodi namijenjeni prehrani dojenčadi od 4. mjeseca starosti, koji uključuju instant pahuljice od riže, instant pahuljice od riže s voćem i instant pahuljice od pšenice uz dodatak probiotičkog soja Bifidobacterium animalis subsp. lactis BB-12® te hrana za posebne medicinske potrebe uz dodatak probiotičkog soja Lacticaseibacillus paracasei subsp. paracasei. Dodatak probiotičkih sojeva u ove proizvode čini ih funkcionalnim pripravcima. Pristunost kulture BB-12® u proizvodima dječje hrane je potvrđena AFLP metodom te PCR-om s početnicama specifičnim za Bifidobcterium vrste. PCR-om za Lacticaseibacillus paracasei vrstu te sekvenciranjem je potvrđena prisutnost tog soja u hrani za posebne medicinske potrebe. Daljnji cilj je bio in vitro ispitati utjecaj ovih različitih matriksa na specifična probiotička svojstva korištenih bakterija. Dječja hrana je imala pozitivan utjecaj tijekom preživljavanja BB-12® u simuliranim uvjetima gastrointestinalnog trakta, u asimilaciji kolesterola, koagregaciji s drugim mikroorganizmima, na antagonističko djelovanje prema drugim Gram-pozitivnim bakterijama ispitano metodom dvostrukog sloja te na antagonističko djelovanje prema bakterijama mliječne kiseline i test-mikroorganizmima ispitano turbidimetrijskom metodom. Hrana za posebne medicinske potrebe je imala pozitivan utjecaj na soj Lacticaseibacillus paracasei subsp. paracasei prilikom asimilacije kolesterola, koagregacije s drugim mikroorganizmima, na antagonističko djelovanje prema test-mikroorganizmima ispitano metodom s rupama u agaru te na inhibicijsko djelovanje prema srodnim bakterijama mliječne kiseline i test-mikroorganizmima ispitano turbidimetrijskom metodom. Adhezija na proteine ekstracelularnog matriksa i Caco-2 stanice, u kombinaciji s antibakterijskom aktivnošću su potvrdile kompetitivnu eksluziju patogena S. Typhimurium FP1 i E. coli 3014 probiotičkim sojevima Bifidobacterium animalis subsp. lactis BB-12® i Lacticaseibacillus paracasei subsp. paracasei.In this research new nutraceutical products intended for infants from 4 months of age have been developed, including instant rice flakes, instant rice flakes with fruit and instant wheat flakes with the addition of probiotic strain Bifidobacterium animalis subsp. lactis BB-12®, and food for special medical purposes with the addition of the probiotic strain Lacticaseibacillus paracasei subsp. paracasei. The addition of probiotic strains to these products makes them functional preparations. The presence of BB-12® culture in baby food products was confirmed by AFLP and PCR with primers specific for Bifidobcterium species. PCR for Lacticaseibacillus paracasei species and sequencing confirmed the presence of this strain in food for special medical purposes. A further aim was to examine in vitro the influence of these different matrices on the specific probiotic properties of the used bacteria. Baby food had a positive effect on the survival of BB-12® in simulated gastrointestinal tract conditions, in cholesterol assimilation, coaggregation with other microorganisms, antagonistic activity against other Gram-positive bacteria tested by the agar spot-test method and antagonistic activity against lactic acid bacteria (LAB) and test-microorganisms tested by turbidimetric method. Food for special medical purposes have had a positive effect on the Lacticaseibacillus paracasei subsp. paracasei during cholesterol assimilation, coaggregation with other microorganisms, antagonistic activity against test-microorganisms tested by agar hole method and inhibitory activity against related LAB and test-microorganisms tested by turbidimetric method. Adhesion to extracellular matrix proteins and Caco-2 cells, combined with antibacterial activity, confirmed the competitive exclusion of S. Typhimurium FP1 and E. coli 3014 pathogens by probiotic strains of Bifidobacterium animalis subsp. lactis BB-12® and Lacticaseibacillus paracasei subsp. paracasei