Fenotypowa i genotypowa charakterystyka probiotycznych szczepów bakteryjnych stosowanych w produktach leczniczych
[Phenotypic and genotypic characterization of probiotic bacterial strains used in medicinal products]

Streszczenie

W pracy ocenie poddano wybrane właściwości szczepów drobnoustrojów określanych jako probiotyki tj. tożsamość, ogólna liczba bakterii oraz wrażliwość na antybiotyki z zastosowaniem opracowanych i zoptymalizowanych procedur badań. Wyniki doświadczeń potwierdziły zgodność badanych właściwości bakterii z deklarowanymi parametrami podanymi na opakowaniu produktu probiotycznego oraz umożliwiły identyfikację ich wrażliwości na antybiotyki. Opracowane metody badań mogą zostać zastosowane do rutynowego
monitorowania cech drobnoustrojów probiotycznych suplementów diety oraz środków spożywczych specjalnego przeznaczenia medycznego zawierających probiotyki.

Abstract

Introduction: The optimization of quality testing strategy of products containing probiotics might allow to general improvement of its safer use in humans. The goal of the study was the evaluation of quality expressed by identity, colony forming unit (CFU) and antibiotic sensitivity of probiotics used in medicinal products available in Poland using the appropriate and validated procedures. Methods: The medicinal products containing L. rhamnosus, L. acidophilus, L. delbrueckii subsp. bulgaricus and B. animalis subsp. lactis, L. helveticus, and L. gasseri were tested for species identity performed with validated rep-PCR (BOXA1R) method. The antimicrobial susceptibility
of working seeds and strains isolated to 26 antibiotics were tested by disk diffusion and E-test methods using relevant references as recommended by EUCAST. The numbers of probiotic strains, expressed as cfu count per package, was done using plating plunge method. Results: All strains tested, except B. lactis, were found to be resistant to trimethoprim-sulphamethoxazole, nalidixic acid, metronidazole, and colistin. B. lactis was resistant to aminoglycosides. L. rhamnosus strains were found to be resistant to vancomycin, (MIC >256μg/ml) similarly to ATCC strains (L. rhamnosus GG 53103 and 244). The sensitivity to other antibiotics was strain specific. The rep-PCR method was found species and strain specific. All products tested fulfilled declared countent as measured by cfu count/package. Conclusions: Quality of medicinal products containing probiotics was found undoubted and confirmed. The optimized strategy of quality monitoring of probiotics used in medicinal products can be used in dietary supplements and foodstuffs intended for particular nutritional uses.