Orally administered Lactobacillus rhamnosus modulates the respiratory immune response triggered by the viral pathogen-associated molecular pattern poly(I:C)

Some studies have shown that probiotics, including Lactobacillus rhamnosus CRL1505, had the potential to beneficially modulate the outcome of certain bacterial and viral respiratory infections. However, these studies did not determine the mechanism(s) by which probiotics contribute to host defense against respiratory viruses. In this work we demonstrated that orally administered Lactobacillus rhamnosus CRL1505 (Lr1505) was able to increase the levels of IFN-γ, IL-10 and IL-6 in the respiratory tract and the number of lung CD3(+)CD4(+)IFN-γ(+) T cells. To mimic the pro-inflammatory and physiopathological consecuences of RNA viral infections in the lung, we used an experimental model of lung inflammation based on the administration of the artificial viral pathogen-associated molecular pattern poly(I:C). Nasal administration of poly(I:C) to mice induced a marked impairment of lung function that was accompanied by the production of pro-inflammatory mediators and inflammatory cell recruitment into the airways. The preventive administration of Lr1505 reduced lung injuries and the production of TNF-α, IL-6, IL-8 and MCP-1 in the respiratory tract after the challenge with poly(I:C). Moreover, Lr1505 induced a significant increase in lung and serum IL-10. We also observed that Lr1505 was able to increase respiratory IFN-γ levels and the number of lung CD3(+)CD4(+)IFN-γ(+) T cells after poly(I:C) challenge. Moreover, higher numbers of both CD103(+) and CD11b(high) dendritic cells and increased expression of MHC-II, IL-12 and IFN-γ in these cell populations were found in lungs of Lr1505-treated mice. Therefore, Lr1505 treatment would beneficially regulate the balance between pro-inflammatory mediators and IL-10, allowing an effective inflammatory response against infection and avoiding tissue damage. Results showed that Lr1505 would induce a mobilization of cells from intestine and changes in cytokine profile that would be able to beneficially modulate the respiratory mucosal immunity. Although deeper studies are needed using challenges with respiratory viruses, the results in this study suggest that Lr1505, a potent inducer of antiviral cytokines, may be useful as a prophylactic agent to control respiratory virus infection.Fil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina. Tohoku University. Graduate School of Agricultural Science. Food Immunology Group; JapónFil: Chiba, Eriko. Tohoku University. Graduate School of Agricultural Science. Food Immunology Group; JapónFil: Tomosada, Yohsuke. Tohoku University. Graduate School of Agricultural Science. Food Immunology Group; JapónFil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Kitazawa, Haruki. Tohoku University. Graduate School of Agricultural Science. Food Immunology Group; JapónFil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin

Dietary supplementation with lactobacilli improves emergency granulopoiesis in protein-malnourished mice and enhances respiratory innate immune response

This work studied the effect of protein malnutrition on the hemato-immune response to the respiratory challenge with Streptococcus pneumoniae and evaluated whether the dietary recovery with a probiotic strain has a beneficial effect in that response. Three important conclusions can be inferred from the results presented in this work: a) protein-malnutrition significantly impairs the emergency myelopoiesis induced by the generation of the innate immune response against pneumococcal infection; b) repletion of malnourished mice with treatments including nasally or orally administered Lactobacillus rhamnosus CRL1505 are able to significantly accelerate the recovery of granulopoiesis and improve innate immunity and; c) the immunological mechanisms involved in the protective effect of immunobiotics vary according to the route of administration. The study demonstrated that dietary recovery of malnourished mice with oral or nasal administration of L. rhamnosus CRL1505 improves emergency granulopoiesis and that CXCR4/CXCR12 signaling would be involved in this effect. Then, the results summarized here are a starting point for future research and open up broad prospects for future applications of probiotics in the recovery of immunocompromised malnourished hosts.Fil: Herrera, Héctor Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Barbieri, Natalia Paola del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina. Universidad Nacional de Chilecito; ArgentinaFil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucuman. Instituto Superior de Investigaciones Biologicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; Argentin

A Novel In-House Enzyme-Linked Immunosorbent Assay for Genotype 3 Hepatitis E Virus Reveals High Seroprevalence in Blood Donors in Northern Argentina

Hepatitis E virus (HEV) is an emergent virus that causes acute hepatitis in immunocompetent hosts and chronic hepatitis in immunocompromised ones. In Latin America, the main circulating genotype (GT) is 3, which is usually of zoonotic origin. Diagnosis and seroprevalence studies mainly rely on the detection of specific antibodies. In Argentina there are scarce data on the seroprevalence of HEV infection mainly due to the lack of awareness of HEV circulation and because HEV testing is not included in routine assays. The following conditions were determined: an optimal antigen concentration of 0.25 µg/ml, a serum dilution of 1:80, gelatin as blocking agent, and a secondary antibody dilution of 1:1000. Specificity (97.8%) and sensibility (100%) were determined using a panel of previously characterized sera. When we compared our in-house ELISA with a commercial test (DIA.PRO), we obtained a kappa index of 0.939. We screened 813 blood donor samples with this newly developed ELISA and found a seroprevalence of 8.31%. We further analyzed the seroprevalence of blood transmitted diseases such as Chagas disease (1.9%), syphilis (1.2%), hepatitis B (0.9%), and brucellosis (0.4%) in a group of 412 blood donors. These samples did not cross reacted with HEV positive samples according to the χ2 test for comparison of proportions. We show for the first time, the circulation of HEV in Tucuman, the most populated city in northern Argentina. We expect that this study will raise the interest of health decision makers who should intercede to include indirect testing of HEV in regular diagnostic protocols. In conclusion, the in-house ELISA developed in this work shows a very good agreement with an already licensed commercial HEV IgG ELISA (DIA.PRO, ITALY), which can be used as an epidemiologic tool for HEV surveillance.Fil: Arce, Lorena Paola. 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. 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; Argentina. Universidad Nacional de Tucumán. Facultad de Medicina; ArgentinaFil: Müller, Melisa Florencia. 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; Argentina. Universidad Nacional de Tucumán. Facultad de Medicina; ArgentinaFil: Martinez, Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaFil: Baiker, Armin. Bavarian Health and Food Safety Authority; AlemaniaFil: Marranzino, Gabriela. Gobierno de la provincia de Tucumán. Ministerio de Salud. Banco Central de Sangre de Tucumán “Dr. César Guerra”; ArgentinaFil: Agote, Felicitas. Gobierno de la provincia de Tucumán. Sistema Provincial de Salud. Banco Central de Sangre "Dr. César Guerra" ; ArgentinaFil: Vizoso Pinto, María Guadalupe. 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; Argentina. Universidad Nacional de Tucumán. Facultad de Medicina; Argentin

Lacticaseibacillus rhamnosus CRL1505 Peptidoglycan Modulates the Inflammation-Coagulation Response Triggered by Poly(I:C) in the Respiratory Tract

Lacticaseibacillus rhamnosus CRL1505 beneficially modulates the inflammation-coagulation response during respiratory viral infections. This study evaluated the capacity of the peptidoglycan obtained from the CRL1505 strain (PG-Lr1505) to modulate the immuno-coagulative response triggered by the viral pathogen-associated molecular pattern poly(I:C) in the respiratory tract. Adult BALB/c mice were nasally treated with PG-Lr1505 for two days. Treated and untreated control mice were then nasally challenged with poly(I:C). Mice received three doses of poly(I:C) with a 24 h rest period between each administration. The immuno-coagulative response was studied after the last administration of poly(I:C). The challenge with poly(I:C) significantly increased blood and respiratory pro-inflammatory mediators, decreased prothrombin activity (PT), and increased von Willebrand factor (vWF) levels in plasma. Furthermore, tissue factor (TF), tissue factor pathway inhibitor (TFPI), and thrombomodulin (TM) expressions were increased in the lungs. PG-Lr1505-treated mice showed significant modulation of hemostatic parameters in plasma (PT in %, Control = 71.3 ± 3.8, PG-Lr1505 = 94.0 ± 4.0, p < 0.01) and lungs. Moreover, PG-Lr1505-treated mice demonstrated reduced TF in F4/80 cells from lungs, higher pro-inflammatory mediators, and increased IL-10 compared to poly(I:C) control mice (IL-10 in pg/mL, Control = 379.1 ± 12.1, PG-Lr1505 = 483.9 ± 11.3, p < 0.0001). These changes induced by PG-Lr1505 correlated with a significant reduction in lung tissue damage. Complementary in vitro studies using Raw 264.7 cells confirmed the beneficial effect of PG-Lr1505 on poly(I:C)-induced inflammation, since increased IL-10 expression, as well as reduced damage, production of inflammatory mediators, and hemostatic parameter expressions were observed. In addition, protease-activated receptor-1 (PAR1) activation in lungs and Raw 264.7 cells was observed after TLR3 stimulation, which was differentially modulated by PG-Lr1505. The peptidoglycan from L. rhamnosus CRL1505 is able to regulate inflammation, the procoagulant state, and PAR1 activation in mice and macrophages in the context of the activation of TLR3 signaling pathways, contributing to a beneficial modulation of inflammation-hemostasis crosstalk.Fil: Zelaya, María Hortensia del Rosario. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Arellano Arriagada, Luciano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Fukuyama, Kohtaro. Tohoku University; JapónFil: Matsumoto, Kaho. Tohoku University; JapónFil: Marranzino, Gabriela. Universidad del Norte Santo Tomás de Aquino. Facultad de Ciencias de la Salud; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Namai, Fu. Tohoku University; JapónFil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Alvarez, Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; ArgentinaFil: Agüero, Graciela. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; ArgentinaFil: Kitazawa, Haruki. Tohoku University; JapónFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin

Stimulation of macrophages by immunobiotic Lactobacillus strains: influence beyond the intestinal tract

Lactobacillus rhamnosus CRL1505 (Lr1505), L. rhamnosus CRL1506 (Lr1506) and L. casei CRL431 (Lc431) are able to stimulate intestinal immunity, but only Lr1505 and Lc431 are able to stimulate immunity in the respiratory tract. With the aim of advancing the understanding of the immunological mechanisms involved in stimulation of distant mucosal sites, this study evaluated the effects of orally administered probiotics on the functions of alveolar and peritoneal macrophages. Compared to a control group, these three lactobacilli were able to significantly increase phagocytic and microbicidal activities of peritoneal macrophages. After intraperitoneal challenge with pathogenic Candida albicans, mice treated with immunobiotics had significantly lower pathogen counts in infected organs. Moreover, lactobacilli- treated mice had a stronger immune response against C. albicans. On the other hand, only Lc1505 and Lc431 were able to improve activity of and cytokine production by alveolar macrophages. Only in these two groups was there better resistance to respiratory challenge with C. albicans, which correlated with improved respiratory immune response. The results of this study suggest that consumption of some probiotic strains could be useful for improving resistance to infections in sites distant from the gut by increasing the activity of macrophages at those sites.Fil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin

Probiotic Lactobacillus strains protect against myelosuppression and immunosuppression in cyclophosphamide-treated mice

This work evaluated the capacity of two probiotic strains, Lactobacillus casei CRL431 and Lactobacillus rhamnosus CRL1506, to protect against myelosuppression and immunosuppression in cyclophosphamide (Cy)-treated mice. Changes inmature granulocytes and progenitor cells in bonemarrow (BM) and blood were studied. In addition, the ability of probiotics to accelerate the recovery of the immune response against the opportunistic pathogen Candida albicans was evaluated.We demonstrated for the first time that the preventive treatmentwith immunomodulatory lactobacilli such as L. casei CRL431 or L. rhamnosus CRL1506 was able to increase immature myeloid progenitors in the BM, allowing an early recovery of myeloid cells after Cy administration. Probiotic lactobacilli were also capable to induce an early recovery of neutrophils in blood, improve phagocytic cells recruitment to infectious sites and increase the resistance against the opportunistic pathogen C. albicans. Although deeper studies regarding the cellular and molecularmechanisms of probiotic actions are needed, these findings support the idea that strains like CRL431 and CRL1506 may accelerate the recovery of Cy-caused immunosuppression by immunopotentiating myeloid cells. Then, probiotic lactobacilli have the potential to be used as alternatives for lessening chemotherapy-induced immunosuppression in cancer patients.Fil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); ArgentinaFil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); ArgentinaFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); ArgentinaFil: Agüero, Graciela. Universidad Nacional de Tucuman. Facultad de Bioquimica, Quimica y Farmacia. Instituto de Bioquimica Clinica Aplicada. Catedra de Bioquimica Clinica I; ArgentinaFil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); Argentina. Universidad Nacional de Tucuman. Facultad de Bioquimica, Quimica y Farmacia. Instituto de Bioquimica Clinica Aplicada. Catedra de Bioquimica Clinica I; Argentin

Morphological characterization of bacteria isolated from surfaces at the "Dr. César Guerra", central blood bank of Tucumán

The microbiomes within the health care field play an important role as reservoirs of infectious diseases. Our goal was to study microbial communities at the Central Blood Bank. The surfaces from the Production and Molecular Biology Services underwent the processes of swabbing and growing in plates (LB pH7). The identification of the isolated strains was carried out through MALDI-TOF, and the morphological characterization was executed by means of Scanning Electron Microscopy (SEM). The species obtained are of clinical importance: as opportunist pathogens (Staphylococcus haemolyticus, Staphylococcus warneri, Clostridium subterminale, Brevibacterium casei, Staphylococcus saprophyticus), multiresistance to antibiotics (Staphylococcus cohnii ssp cohnii), formation of spores (Bacillus cereus group, Bacillus sp), or responsible for serious infections related to biofilms (Micrococcus luteus, Burkholderia cenocepacia, Pseudomonas stutzeri). Some isolated bacteria are of significance to bioremediation (Dietzia cinnamea, Dietzia maris, Bacillus subtilis/amyloliquefaciens/vallismortis, Pseudomonas stutzeri; Bacillus altitudinis/pumilus), and production of enzymes (Bacillus megaterium). The analysis of some strains with SEM revealed the presence of tubular structures, possibly nanotubes, associated to the production of biofilms and resistance to antimicrobials. Our findings show the bacterial diversity predominant at the blood bank, and describe the structures possibly involved in mechanisms of pathogenicity.Fil: D'arpino, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alvarado, Natalia Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Galván, Fátima Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Martinez, L.. No especifíca;Fil: Albarracín, Virginia Helena. Universidad Nacional de Tucumán. Facultad de Agronomía y Zootecnia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaXXXVIII Jornadas Científicas de la Asociación de Biología de TucumánSan Miguel de TucumánArgentinaAsociación de Biología de Tucumá

Non-viable immunobiotic Lactobacillus rhamnosus CRL1505 and its peptidoglycan improve systemic and respiratory innate immune response during recovery of immunocompromised-malnourished mice

The effect of non-viable Lactobacillus rhamnosus CRL1505 and its cell wall and peptidoglycan on respiratory immunity in malnourished mice was studied.Weaned mice were malnourished with a protein-free diet for 21d and received BCD during 7d (BCD) or BCD with nasal non-viable L. rhamnosus CRL1505 (BCD + UV) or its cell wall (BCD + CW) or peptidoglycan (BCD + PG) supplementation during last 2d of the treatment. Malnourished mice without treatment (MNC) and well-nourished mice (WNC) were used as controls. Mice were infected nasally with Streptococcus pneumoniae after treatments. Resistance against pneumococci was reduced in MNC mice.Repletion with BCD reduced lung and blood bacterial cell counts when compared to MNC mice but the counts did not reach the levels of the WNC group. However, when malnourished mice received BCD + UV, BCD + CW or BCD+PG, pneumococci was not detected in lung or blood samples. Pneumococcal infection increased the levels of TNF-α, IL-1β, IL-6, and IL-10 in the respiratory tract, however the values were lower in MNC than in WNC mice.BCD + UV and BCD + PG groups showed values of phagocytes, IL-1β and IL-6 that were similar to WNC mice,while TNF-α was significantly higher in those groups when compared to WNC mice. Moreover, BCD + UV and BCD+PG treatments improved levels of respiratory IL-10, reaching values that were superior to those observed in WNC mice. The work demonstrates for the first time that non-viable probiotic bacteria or their cellular fractions could be an interesting alternative as mucosal mmunomodulators, especially in immunocompromised hosts in which the use of live bacteria might be dangerous.Fil: Kolling, Yanina Noralí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentin

Scanning electron microscopy for the study of microbiomes on surfaces of the medical-assistance field

El objetivo de este trabajo fue identificar y caracterizar las comunidades microbianas presentes en el ámbito médico-asistencial. Se tomaron muestras por duplicado mediante cinta de papel e hisopado y siembra en placas con medio de cultivo LB pH7 adicionado con antibióticos Cicloheximida (CH) y Ciclohehimida más Ácido Nalidixico (CH/NA). Los sitios de muestreo fueron: mesadas, aire acondicionado y equipamientos, de los Servicios de Producción, Distribución y Biología Molecular del Banco Central de Sangre Dr. César Guerra, (PRIS-SI.PRO.SA). El análisis de las cintas aplicando microscopía electrónica de barrido (SEM, Scannig Electron Microscopy) permitió detectar biofilms en los diferentes sitios de muestreo. Los mismos presentaron una organización tridimensional compleja caracterizada por microorganismos dispuestos en capas inmersas en abundante material extracelular. Los cultivos permitieron aislar 45 colonias (gram + y gram ) que exhibieron diversos morfotipos (cocos, bacilos y cocobacilos) de mesadas, aire acondicionado y equipos. Mediante SEM fue posible analizar en detalle la estructura topográfica, organización y morfología de las bacterias aisladas. Se observó además, que algunas colonias establecían contactos estrechos. SEM reveló un amplio espectro de relaciones entre ellas. Fue posible poner en evidencia cambios morfológicos en las bacterias como también la presencia de gran cantidad de material extracelular en los sitios de interacción. También, se identificaron diferencias en la conformación ultraestructural de distintos sectores de algunas colonias. Este trabajo propone a las técnicas de microscopía de alta resolución como herramientas claves para el estudio in-situ de biofilms en superficies del ámbito médico-asistencial, dada la estrecha la estrecha relación que presentan con las enfermedades humanas.Fil: D'arpino, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Galván, Fátima Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Alvarado, Natalia Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Martinez, L.. No especifíca;Fil: Albarracín, Virginia Helena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaX Jornadas de Investigación y Desarrollo 2020San Miguel de TucumánArgentinaUniversidad de San PabloInstituto de Desarrollo e Innovación Tecnológica para la Competitividad Territoria

Estudio de microbiomas en ámbitos médicos-asistenciales por microscopía electrónica de barrido

The identification of microbiomes present in the health field is extremely important given the close relationship with human diseases. The aim of this study was to investigate the composition of microorganisms on the laboratory surfaces of the Central Blood Bank "Dr. César Guerra", (PRIS-SI.PRO.SA). Duplicate samples from the countertops, air conditioners and the equipment on the Production, Distribution and Molecular Biology services were taken using paper tape and swabs. Plating was carried out in LB pH7 culture medium containing Cycloheximide (CH) and Cyclohehimide/ Nalidixic Acid (CH / NA) antibiotics. Both the paper tape and the colonies obtained from the cultures were subjected to Scanning Electron Microscopy (SEM). Sampling using paper tape allowed the presence of microbial biofilms to be detected in the internal part of the Production service centrifuge, in the Distribution service platelet shaker and in the Molecular Biology service countertops. They presented a complex three-dimensional organization characterized by microorganisms of different morphology arranged in layers immersed in abundant extracellular material. The tape analysis also revealed the presence of isolated bacteria (cocci and bacilli) or the formation of small groups of them at the different sampling sites. The cultures allowed isolating predominant microorganisms from countertops, air conditioners and equipment. A total of 45 colonies (gram + and gram -) that exhibited various morphotypes (cocci, bacilli and cocobacilli) were isolated. Using SEM, it was possible to analyze in detail the structure, organization and morphology of the bacteria in culture. Furthermore, it was observed that many colonies established close contacts. The SEM study revealed a wide spectrum of relationships among them. It was possible to analyze the contact points among interacting colonies, revealing morphological changes in the bacteria as well as a large amount of extracellular material at the interaction sites. In addition, the topographic analysis of the colonies showed differences in the conformation of the different sectors in some of them. This work, aimed at analyzing the microbiological communities developed in situ in healthcare settings, proposes high-resolution microscopy techniques as key tools for the study in situ of biofilms on a surface, which study is lacking in our country.Fil: D'arpino, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Galván, Fátima Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Alvarado, Natalia Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Marranzino, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Martinez, L.. Centro Integral de Microscopía Electrónica; ArgentinaFil: Albarracín, Virginia Helena. Universidad Nacional de Tucumán. Facultad de Agronomía y Zootecnia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaIV Reunión Conjunta de Sociedades de Biología de la República ArgentinaTucumánArgentinaSociedad de Biología de TucumánSociedad de Biología de CuyoSociedad de Biología de Córdob