Antibiotic resistance and probiotics properties of lactic acid bacteria isolated from Andean vegetable and animal products

In Andean regions of America, many traditional fermented foods and beverages are produced. Vegetable traditional products involve quinoa and amaranth whose widespread consumption as grains and fermented flours (sourdoughs) has grown recently due to their high nutritional value and lack of gluten. Fermented potatoes (Tocosh) have also been produced from ancient times in the Central Andean region of Peru, whereas llama meat fermented sausages are traditionally produced and consumed in Northwestern Argentina. At present, there is great concern that commensal bacterial populations from food and the gastrointestinal tract (GIT) of humans and animals, such as lactic acid bacteria (LAB) can serve as reservoir for antibiotic resistance genes, which can be horizontally transferred to other microorganisms. On these bases, the antibiotic resistance profiles (EFSA-2012) of eighty LAB strains isolated from Andean products were investigated. The analyzed exhibited a general sensitive profile against ampicilline (AMP), vancomycin (VAN), gentamycin (GEN), kanamycin (KAN), streptomycin (STR), erythromycin (ERY), clindamycin (CLI), tetracycline (TET) and chloramphenicol (CHL). Several strains were resistant mainly to the aminoglycosides (KAN, STR and GEN) while a multi-resistant pattern was found particularly among strains isolated from quinoa /amaranth grains and sourdoughs. However, sensitive LAB strains were observed among Lb. sakei, Lb. reuteri, Lb. plantarum, Lb. casei and Leuc. mesenteroides. On the other hand, to select LAB strains with probiotic potential the resistance to stress factors (lysozyme, bile salts and low pH) present in the GIT, surface properties (hydrophobicity, autoaggregation and adhesion to caco-2 cells) and production of antimicrobial compounds were evaluated. Results allowed selecting Lb. plantarum (2 strains), Lb. reuteri (1) and Lb. casei (1) as the best candidates to be used as probiotics.Fil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Vera Pingitore, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Fontana, Cecilia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Università Cattolica del Sacro Cuore; ItaliaFil: Cocconcelli, Pier. S.. Università Cattolica del Sacro Cuore; ItaliaFil: Jimenez, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Plumed Ferrer, Carmen. University Of Eastern Finland.; FinlandiaFil: von Wright, Atte. University Of Eastern Finland.; FinlandiaFil: Aznar, Rosa. Universidad de Valencia; España11th International Symposium on Lactic Acid Bacteria: Health, Sustainability, Diversity, and ApplicationEgmond aan ZeePaíses BajosRoyal Netherlands Society for MicrobiologyFederation of European Microbiological Societie

Elevated Humoral Immune Response to SARS-CoV-2 at High Altitudes Revealed by an Anti-RBD “In-House” ELISA

The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused a global pandemic with dramatic health and socioeconomic consequences. The Coronavirus Disease 2019 (COVID-19) challenges health systems to quickly respond by developing new diagnostic strategies that contribute to identify infected individuals, monitor infections, perform contact-tracing, and limit the spread of the virus. In this brief report, we developed a highly sensitive, specific, and precise “In-House” ELISA to correctly discriminate previously SARS-CoV-2-infected and non-infected individuals and study population seroprevalence. Among 758 individuals evaluated for anti-SARS-CoV-2 serology in the province of Tucumán, Argentina, we found a weak correlation between antibodies elicited against the RBD, the receptor-binding domain of the Spike protein, and the nucleocapsid (N) antigens of this virus. Additionally, we detected mild levels of anti-RBD IgG antibodies in 33.6% of individuals diagnosed with COVID-19, while only 19% showed sufficient antibody titers to be considered as plasma donors. No differences in IgG anti-RBD titers were found between women and men, neither in between different age groups ranging from 18 to 60. Surprisingly, individuals from a high altitude village displayed elevated and longer lasting anti-RBD titers compared to those from a lower altitude city. To our knowledge, this is the first report correlating altitude with increased humoral immune response against SARS-CoV-2 infection.Fil: Tomas Grau, Rodrigo Hernán. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Ploper, Diego. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Avila, Cesar Luis. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Vera Pingitore, Esteban. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Maldonado Galdeano, María Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Chaves, Analia Silvina. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Socias, Sergio Benjamin. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Stagnetto, Agustín. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Navarro, Silvia Adriana. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; ArgentinaFil: Chahla, Rossana Elena. Gobierno de la Provincia de Tucumán. Ministerio de Salud; ArgentinaFil: Aguilar López, Mónica. Gobierno de la Provincia de Tucuman. Hospital de Dia Presidente Nestor Carlos Kirchner; ArgentinaFil: Llapur, Conrado Juan. Gobierno de la Provincia de Tucumán. Ministerio de Salud; ArgentinaFil: Aznar, Patricia. Gobierno de la Provincia de Tucuman. Hospital de Dia Presidente Nestor Carlos Kirchner; ArgentinaFil: Alcorta, María Elena. Gobierno de la Provincia de Tucuman. Hospital de Dia Presidente Nestor Carlos Kirchner; ArgentinaFil: Costas, Dardo. Gobierno de la Provincia de Tucuman. Hospital de Dia Presidente Nestor Carlos Kirchner; ArgentinaFil: Flores, Isolina. Gobierno de la Provincia de Tucuman. Hospital de Dia Presidente Nestor Carlos Kirchner; ArgentinaFil: Heinze, Dar. University of Boston. School of Medicine; Estados UnidosFil: Apfelbaum, Gabriela. Universidad Nacional de Tucumán; ArgentinaFil: Mostoslavsky, Raul. Harvard Medical School; Estados UnidosFil: Mostoslavsky, Gustavo. Harvard Medical School; Estados UnidosFil: Cazorla, Silvia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Perdigon, Gabriela del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Chehin, Rosana Nieves. Universidad Nacional de Tucumán. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucumán. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Investigaciones en Medicina Molecular y Celular Aplicada del Bicentenario; Argentin

Long-term analysis of antibodies elicited by SPUTNIK V: A prospective cohort study in Tucumán, Argentina

Background: Gam-COVID-Vac (SPUTNIK V) has been granted emergency use authorization in 70 nations and has been administered to millions worldwide. However, there are very few peer-reviewed studies describing its effects. Independent reports regarding safety and effectiveness could accelerate the final approval by the WHO. We aimed to study the long-term humoral immune response in nay¨ve and previously infected volunteers who received SPUTNIK V. Methods: Humoral immune responses, assayed by anti-SARS-CoV-2-spike-RBD IgG ELISA and neutralization assays, were measured in 602 healthcare workers at 0, 14, 28, 60 and 180 days after receiving SPUTNIK V between December 2020 and July 2021 in Tucuman, Argentina. Findings: Seroconversion was detected in 97% of individuals after 28 days post-vaccination (dpv) (N = 405). Anti-RBD titers began to decrease after 60 dpv (N = 328), but remained detectable in 94% at 90 dpv (N = 224). At 180 dpv, anti-RDB titers persisted in 31% (N = 146). Previous infection triggered an increased immune response to the first dose and increased neutralization activity against variants of concern (VOC). Second doses in previously infected individuals further increased titers, even 90 dpv (N = 75). Basal antibody titers had more influence on postvaccination anti-RBD responses than the time elapsed between diagnosis and vaccination (N = 274). Interpretation: Data presented herein provides essential knowledge regarding the kinetics of antibodies induced by SPUTNIK V up to six months after immunization, and suggests that when considering one-dose vaccination policies for individuals with previous SARS-CoV-2 infection, serological studies to determine basal titers may be important, independent of when diagnosis occurred.Fil: Chahla, Rossana Elena. Ministerio de Salud Pública de Tucumán; ArgentinaFil: Tomas Grau, Rodrigo Hernán. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; ArgentinaFil: Cazorla, Silvia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Ploper, Diego. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; ArgentinaFil: Vera Pingitore, Esteban. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; ArgentinaFil: Aguilar López, Mónica. Gobierno de la Provincia de Tucuman. Ministerio de Salud. Departamento Bioquimico. Laboratorio de Salud Publica.; ArgentinaFil: Aznar, Patricia. Gobierno de la Provincia de Tucuman. Ministerio de Salud. Departamento Bioquimico. Laboratorio de Salud Publica.; ArgentinaFil: Alcorta, María Elena. Gobierno de la Provincia de Tucuman. Ministerio de Salud. Departamento Bioquimico. Laboratorio de Salud Publica.; ArgentinaFil: Velez, Eva Maria del Mar. Gobierno de la Provincia de Tucuman. Ministerio de Salud. Departamento Bioquimico. Laboratorio de Salud Publica.; ArgentinaFil: Stagnetto, Agustín. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; ArgentinaFil: Avila, Cesar Luis. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; ArgentinaFil: Maldonado Galdeano, María Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Socias, Sergio Benjamin. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; ArgentinaFil: Heinze, Dar. University Of Boston. School Of Medicine. Center For Regenerative Medicine.; Estados UnidosFil: Navarro, Silvia Adriana. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; ArgentinaFil: Llapur, Conrado Juan. Ministerio de Salud Pública de Tucumán; ArgentinaFil: Costas, Dardo. Gobierno de la Provincia de Tucuman. Ministerio de Salud. Departamento Bioquimico. Laboratorio de Salud Publica.; ArgentinaFil: Flores, Isolina. Gobierno de la Provincia de Tucuman. Ministerio de Salud. Departamento Bioquimico. Laboratorio de Salud Publica.; ArgentinaFil: Edelstein, Alexis. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; ArgentinaFil: Kowdle, Shreyas. Icahn School of Medicine at Mount Sinai; Estados UnidosFil: Perandones, Claudia. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; ArgentinaFil: Lee, Benhur. Icahn School of Medicine at Mount Sinai; Estados UnidosFil: Apfelbaum, Gabriela. Universidad Nacional de Tucumán; ArgentinaFil: Mostoslavsky, Raul. Harvard Medical School; Estados UnidosFil: Mostoslavsky, Gustavo. University Of Boston. School Of Medicine. Center For Regenerative Medicine.; Estados UnidosFil: Perdigon, Gabriela del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Chehin, Rosana Nieves. Universidad Nacional de Tucuman. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Gobierno de la Provincia de Tucuman. Ministerio de Salud. Sistema Provincial de Salud. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto de Investigaciones En Medicina Molecular y Celular Aplicada del Bicentenario.; Argentin

Effect of lyophilization and storage temperature on the activity of salivaricin CRL 1328, a potential bioactive ingredient of a urogenital probiotic product

Bacteriocins are antimicrobial peptides with potential applications as therapeutic agents for the treatment of microbial infections. The aim of this work was to investigate the effect of different protectors on the activity of salivaricin CRL1328, a bacteriocin produced by Lactobacillus salivarius CRL1328, during the lyophilization process and subsequent storage at different temperatures for 18 months using statistical models. Different protectors such as mannitol, Tween 80, polyethylene glycol 8000 (PEG), monosodium glutamate (MSG), reconstituted skim milk, sucrose and ascorbic acid were used for the lyophilization and storage of salivaricin. The biplot of principal component analysis was used for the interpretation of the interactions between the different factors studied. The antimicrobial activity of salivaricin was dependent mainly on temperature, and also on the time of storage and protector assayed. The stability of salivaricin was higher at -20°C and 4°C than 25°C and decreased during the time of storage; however, salivaricin was active after 18 months of storage at 25°C. Sucrose, mannitol plus sucrose, PEG plus sucrose and MSG were the most effective agents in protecting the bacteriocin during the lyophilization process. Effective maintenance of the activity of the bacteriocin was observed by storage with sucrose and ascorbic acid at –20°C as well as with PEG plus sucrose at 4°C and –20°C. The results obtained suggest that sucrose alone or combined with PEG can effectively maintain the activity of salivaricin during lyophilization and storage. This study provides useful information for the potential application of salivaricin as a bioactive principle for a pharmaceutical formulation.Fil: Vera Pingitore, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Bru Chauve, Elena Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Nader, Maria Elena Fatima. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin

Potential applications of bacteriocins as bioactive substances for human health

Lactic acid bacteria are constituents of most of the indigenous microbiota or mucosarelated ecosystems, both in humans or animals. During the last years, many bacteriocins produced by bacteria classified into this group were described. Bacteriocins are ribosomally-synthesized antimicrobial peptides produced by a wide range of bacteria. Historically, they were used as bioactive compounds to prevent the growth of pathogen or spoilage microorganisms in foods. The most widely used is nisin, produced by Lactococcus lactis, that is included in the CODEX and has the GRAS (Generally Recognized as Safe) status to be incorporated into different types of processed or longterm storage foods. As bacteriocins are active against many pathogens, potentially pathogens or some drug-resistant pathogens, some of them have been thought as alternative bioactive substances to avoid the broad side-effects and alarming resistance dissemination produced by the use of classical antibiotics. Our research group is working in the design of a beneficial product to prevent prevent women urogenital infections, and we were the first to describe salivaricin CRL 1328, a bacteriocin produced by Lactobacillus salivarius isolated from human vagina. This bioactive compound is heat-stable, and active against a wide variety of urogenital pathogens. The molecular characterization of salivaricin indicates that is constituted by two structural peptides and the evaluation of the mechanism of action reveals that acts by dissipating the proton motive force of the sensitive pathogens. As we are interested to incorporate salivaricin into a pharmaceutical product, the optimization of the culture media and physico-chemical conditions for the higher production were determined. And also the stability to the freeze-drying process and to the long term storage conditions was studied. Some of these experiments were performed by the application of statistical methods that allowed a faster and easier interpretation of the experimental results obtained. The incorporation of bacteriocins as bioactive compounds in a pharmaceutical product needs different type of studies to demonstrate the no existence of adverse effects, which must be performed both in in vitro and in in vivo experimental systems. Most of the bacteriocins do not exert significant cytoxicity or hemolytic activity against human erythrocytes in in vitro assays. Experiments in animal models have shown the success of some lantibiotics for the treatment of infections produced by Streptococcus pneumoniae and methicillin-resistant Staphylococcus aureus (MRSA), and also in the prevention of gingivitis. Nisin is included into a pharmaceutical product to prevent cow mastitis, and is proposed as spermicidal for human use. In this chapter, a revision of many of the bacteriocins described for prevention of human health are reviewed, together with those available commercially, evaluated by in vitro or in vivo assays. Up today, the use of bacteriocins as bioactive compounds into pharmaceutical products for human use has not been accepted yet by the regulators or reference organizations. There are a wide variety of bacteriocins produced by different bacterial genera that must be further studied. More complex experiments are needed to encourage and support the inclusion of these types of bioactive substances into products that could be applied to prevent or treat many of the wide arrays of pathogenic microorganisms that affect human and animal health.Fil: Nader, Maria Elena Fatima. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Vera Pingitore, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Juárez Tomás, María Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin

Comparative proteomic analyses for elucidating metabolic changes during EPS production under different fermentation temperatures by Lactobacillus plantarum Q823

Exopolysaccharide (EPS)-producing bacteria are of growing interest in industrial processes, mainly concerning food. Lactic acid bacteria are widely appreciated for their GRAS (generally recognized as safe) status and their ascertained or putative probiotic features. Detailed investigation on what happens at metabolic level during EPS production is scarce in the literature. The facultative heterofermenter Lactobacillus plantarum Q823 was studied in order to compare growth and EPS production at 30 °C and 37 °C. A higher growth rate was observed at 37 °C, whereas, a significantly higher (tenfold increase) EPS amount was produced at 30 °C. To understand the molecular mechanisms leading to the different EPS production in the two conditions, a comparative proteomic experiment was performed. The results of the in-gel proteomics revealed that: i) at 37 °C a higher abundance of proteins involved in carbon catabolism and nucleic acid biosynthesis together with a significant amount of stress proteins was observed; ii) at 30 °C the production of an atypical manganese-containing non-heme catalase (pseudocatalase) was increased, in agreement with previous data reporting that growth-rates of catalase negative Lactobacillus plantarum strains were greater than that of catalase positive strains. Taken together, all these findings provide further insights about the metabolic pathways stimulated during EPS production, and the mechanism that triggers EPS biosynthesis.Fil: Vera Pingitore, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina. Università di Torino; ItaliaFil: Pessione, Alessandro. Università di Torino; ItaliaFil: Fontana, Cecilia Alejandra. Università Cattolica del Sacro Cuore; Italia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mazzoli, Roberto. Università di Torino; ItaliaFil: Pessione, Enrica. Università di Torino; Itali

Antibiotic resistance assessment of lactic acid bacteria isolated from traditional Andean fermented sausages

Antimicrobial susceptibility profiles of lactic acid bacteria (LAB) from different foods, including fermented dry sausages have been described. Several studies suggest that the presence of antimicrobial resistances in the microbial communities of fermented foods is a consequence of the use of antibiotics in animal farming. In this study we have analyzed the antimicrobial resistance profiles of LAB from Andean traditional fermented sausages, produced in areas characterized by a low anthropic impact. These artisanal fermented sausages produced in the northern province of Jujuy, Argentina are usually prepared with 75% of llama meat, 19% of pork fat, together with salt, sodium nitrite, sucrose and species (white pepper, clove and garlic). A total of 48 Lactobacillus sakei strains isolated from llama fermented sausages presenting different RAPD profiles were selected for the antimicrobial susceptibility tests. All the antibiotics recommended for the EFSA (2012) were included in the assay: ampicillin and vancomycin as inhibitors of cell wall synthesis, clindamycin, chloramphenicol, erythromycin, gentamicin, kanamycin, streptomycin and tetracycline as inhibitors of protein synthesis. The minimum inhibitory concentration of antibiotics (MICs, µg/mL) for all bacteria was determined by the broth microdilution method (International Standard [ISO 10932:2010(E)]. Differences in the antimicrobial susceptibility were observed among the strains. All of them were sensitive to erythromycin, tetracycline and clindamycine. Resistant to ampicillin was observed only in a multiresistant strain presenting a resistant profile to four antibiotics. Most of the strains were resistant to the aminoglycosides, mainly to streptomycin (22 strains) but also to kanamycin (5 strains) and gentamycin (5 strains). The rest of the strains were sensitive to all the antibiotics tested. This data indicates that also in marginal areas the resistance to clinically relevant antibiotics is widespread in the adventitious bacterial communities responsible for meat fermentation, although the high prevalence of aminoglycoside resistance may be related to intrinsic resistance mechanisms.Fil: Fontana, Cecilia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Università Cattolica del Sacro Cuore; ItaliaFil: Vera Pingitore, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Cocconcelli, Pier S.. Università Cattolica del Sacro Cuore; Italia2nd International Symposium on Fermented MeatValenciaEspañaConsejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimento

Design of novel urogenital pharmabiotic formulations containing lactobacilli, salivaricin CRL 1328 and non-microbial compounds with different functionalities

Context: The administration of pharmabiotics is a promising alternative to antimicrobial drugs for the treatment and/or prevention of female urogenital infections. Objective: To design pharmabiotic formulations including bioactive ingredients of microbial origin combined with non-microbial substances and then to evaluate the stability of the combinations during freeze-drying and storage. Materials and Methods: Different formulations including Lactobacillus gasseri CRL 1263, Lactobacillus salivarius CRL 1328, salivaricin CRL 1328 (a bacteriocin) and non-microbial compounds (lactose, inulin and ascorbic acid) were assayed and the ingredients were freeze-dried together or separately. The formulations were stored in gelatin capsules at 4ºC for 360 days. Results: The viability of lactobacilli was affected to different extents depending on the strains and on the formulations assayed. L. salivarius and ascorbic acid were successfully combined only after the freeze-drying process. Salivaricin activity was not detected in formulations containing L. gasseri. However, when combined with ascorbic acid, lactose, inulin or L. salivarius, the bacteriocin maintained its activity for 360 days. The selected microorganisms proved to be compatible for their inclusion in multi-strain formulations together with lactose, inulin and ascorbic acid. Salivaricin could be included only in a L. salivarius CRL 1328 single-strain formulation together with non-microbial substances. Conclusions: This study provides new insights into the design of urogenital pharmabiotics combining beneficial lactobacilli, salivaricin CRL 1328 and compounds with different functionalities.Fil: Vera Pingitore, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Juárez Tomás, María Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Weise, Birgitt. Hannover Medical School. Institute of Biometry; AlemaniaFil: Nader, Maria Elena Fatima. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin