INVESTIGADORES
VERA PINGITORE Esteban
congresos y reuniones científicas
Título:
DESIGN OF A NEW UROGENITAL PROBIOTIC FORMULA CONTAINING SALIVARICIN, LACTOBACILLI AND OTHER BIOACTIVE SUBSTANCES
Autor/es:
VERA PINGITORE E.; JUÁREZ TOMÁS M. S.; WEISE B.; NADER-MACIAS M.E.
Lugar:
Córdoba
Reunión:
Congreso; 1ª Reunión Internacional de Ciencias Farmacéuticas; 2010
Institución organizadora:
Universidad Nacional de Córdoba
Resumen:
DESIGN OF A NEW UROGENITAL PROBIOTIC FORMULA CONTAINING SALIVARICIN, LACTOBACILLI AND OTHER BIOACTIVE SUBSTANCES Vera Pingitore E1, Juárez Tomás MS1, Wiese B2, Nader-Macías ME1 1. Centro de Referencia para Lactobacilos (CERELA–CONICET) Chacabuco 145 (T4000ILC), San Miguel de Tucumán, Tucumán, Argentina. 2. Institute of Biometry, Hanover Medical School, Germany Introduction. The urogenital infections (UGI) affect over 1 billion women worldwide each year (1). The conventional treatments are based in prescribed antibiotic therapies. However, they produce an imbalance of indigenous microbiota, adverse effects and antibiotic resistance, being not recommended for pregnant women. A promising alternative for the UGI is the administration of probiotics, defined as “live microorganisms which, when administered in adequate amounts, exert a beneficial physiological effect in the host health” (2). The main objective of our research group is the design of novel probiotic pharmaceutical products containing lactobacilli strains combined with antagonistic substances, as for example bacteriocins, which are antimicrobial peptides ribosomally synthesized by some bacteria. Salivaricin CRL1328 is a bacteriocin produced by Lactobacillus salivarius CRL 1328, a strain isolated from healthy human vagina, with potential applications for preventing urogenital infections (3,4). Salivaricin is active against pathogens including Gardnerella vaginalis, Enterococcus faecalis, E. faecium, Listeria monocytogenes, Streptococcus agalactiae, Staphylococcus sapropyticus and Neisseria gonorrheae. The long term storage of microorganisms is a challenge in the pharmaceutical area. Lyophilization is one of the process usually employed and the carefully selection of lyoprotectors plays a major role in the stability of bioactive compounds. The aim of this work was to evaluate the resistance of beneficial lactobacilli combined with salivaricin and other bioactive substances to the freeze-drying, and their stability during their shelf-life Materials and methods. Lactobacillus salivarius CRL 1328 and Lactobacillus gasseri CRL 1263 were grown in LAPTg medium at 37ºC. Sixteen different combinations of bacteria, salivaricin, lactose, inulin and ascorbic acid were designed and evaluated. Later, based on these results, a new set of eight combinations was assayed. The samples were lyophilized and stored in gelatin capsules at 4ºC during 180 days taking samples periodically. The viability of the strains (CFU/g), as well as salivaricin activity (AU/g) were evaluated to determine the resistance to the lyophilization process and during the storage. The data were statistically analyzed by applying ANOVA test and a model proposed to evaluate the experimental results obtained. Results. The selected microorganisms were compatible between them and with salivaricin. The ANOVA analysis showed that the protective agents and the storage time were the two factors that affect both, the activity of salivaricin and the viability of Lactobacillus strains. The viability of both strains was higher when lactose was present in the mixture. L.g.CRL 1263 was highly conserved with ascorbic acid or lactose, but not with inulin assayed individually. The negative effect of inulin disappeared when the three substances were present in the mixture. Ascorbic acid was detrimental for L.s.CRL 1328 when lyophilized simultaneously. However, when added after freeze-drying process, the survival rate increased significantly. The antimicrobial activity of salivaricin was lost in the samples combined with L.g.CRL 1263, probably by its adsortion to the bacterial cell. A progressive reduction of salivacin activity during storage was detected in the samples with inulin. However, when combined with lactose, ascorbic acid or L.s.CRL 1328, the activity was maintained for 180 days. Conclusions. This is the first study that provides new information for the potential application of salivaricin as bioactive principle, alone or combined with probiotic microorganisms, for their inclusion in probiotic pharmaceutical formulations. References. 1. Reid G. Probiotic Lactobacilli for urogenital health in women. J Clin Gastroenterol. 2008 Sep;42(2): 234-6. 2. Reid G, Sanders ME, Gaskins HR, Gibson GR, Mercenier A, Rastall R, et al. New scientific paradigms for probiotics and prebiotics. J Clin Gastroenterol. 2003 Aug;37(2):105-18. 3. Ocaña VS, Pesce de Ruiz Holgado A, Nader-Macías ME. Characterization of a bacteriocin-like substance produced by a vaginal Lactobacillus salivarius strain. Appl Environ Microbiol. 1999 Dec;65(12):5631-5. 4. Vera Pingitore E, Hébert EM, Nader-Macías ME, Sesma F. Characterization of salivaricin CRL 1328, a two-peptide bacteriocin produced by Lactobacillus salivarius CRL 1328 isolated from the human vagina. Res Microbiol. 2009 Jul-Aug;160(6):401-8.