Immunological effects of yogurt addition to a re-nutrition diet in a malnutrition experimental model

The therapeutic and preventive effects of yogurt and lactic acid bacteria on diseases such as cancer, infection and gastrointestinal disorders are well documented. The aim of this research was to study the effects of different doses of yogurt addition after milk re-nutrition diet, on the recovery of the intestinal barrier and mucosal immune function. Experiments were performed on groups of mice, malnourished and re-nourished with milk during 7 d, and mice with diet supplemented with yogurt for 2, 5 and 7 consecutive d. Nutritional parameters such as weight gain, serum total protein, and the number of IgA, IgM and IgG B cells of the small intestine were determined. We also quantified intraepithelial leukocytes, mastocytes and goblet cells, and performed structural and ultrastructural studies on the small intestine. We observed that 5 d of yogurt feeding was the optimal dose for improving gut barrier function and mucosal immune system in a malnutrition model. This effect was not observed with milk re-nutrition. Although the results were better for 5 d of yogurt, addition for 7 d also showed beneficial effects. Yogurt feeding in our model did not impair any gut functions. These results suggest that yogurt addition after a re-nutrition diet gives better recovery of intestinal function than the re-nutrition diet usually recommended. Although these results were obtained in an animal model, they indicate that consumption of yogurt by malnourished children might accelerate the restoration of gut function.Fil: Gauffin Cano, María Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad del Norte Santo Tomás de Aquino; ArgentinaFil: Aguero Villoldo, Maria Graciela. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; ArgentinaFil: Perdigon, Gabriela del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentin

Yoghurt accelerates the recovery of defence mechanisms against Streptococcus pneumoniae in protein-malnourished mice

Experiments studied the effect of yoghurt on the recovery of defence mechanisms against Streptococcus pneumoniae respiratory infection in malnourished mice. Weaned mice were malnourished with a protein-free diet (PFD) for 21d. Malnourished mice were made replete with a balanced diet (BD), yoghurt, or the BD with supplemental yoghurt (BD + Y) for 7, 14 or 21d. The normal control (NC) group was fed the BD whereas malnourished control (MC) mice consumed only the PFD. Mice were challenged with pneumococci at the end of each dietary treatment. MC mice showed increased susceptibility to pneumococcal infection. Blood leucocytes, phagocyte activity and serum and bronco-alveolar anti-pneumococcal IgG and IgA were significantly lower in the MC than in the NC group. Repletion of malnourished mice with the BD for 21d was necessary to obtain a response to infection similar to that of NC mice; however, administration of the BD + Y for 14d was enough to normalise the immune defence mechanisms. Histological examination of MC lungs showed progressive loss of alveolar architecture. Lung injuries were significantly less pronounced in NC mice. Mice treated with the BD + Y for 14d showed histological signs similar to the NC group. The present study showed that administration of yoghurt to malnourished mice induced an early recovery of the immunological parameters studied. Despite the uncertainties about the mechanisms involved and about the human relevance of the effects observed in animal models, the present study provides a strong rationale for the hypothesis that yoghurt consumption by malnourished hosts will accelerate the recovery of the immune mechanisms involved in the protection against respiratory infections.Fil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Racedo, Silvia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Aguero Villoldo, Maria Graciela. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Alvarez, Gladis Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentin

Evaluación de las propiedades adyuvantes de fracciones celulares de bacterias inmunobióticas para el diseño de nuevas formulaciones vacunales contra Streptococcus pneumoniae

Streptococcus pneumoniae (neumococo) representa uno de los mayores problemas de salud a nivel mundial debido a que es causa común de infecciones del tracto respiratorio y enfermedades invasivas especialmente en grupos de alto riesgo como niños en sus primeros años de vida, adultos y pacientes inmunocomprometidos. La emergencia de cepas de neumococos multiresistentes en el mundo ha conducido a un incremento en la necesidad de la prevención de infecciones por S. pneumoniae mediante la vacunación. Sin embargo, la vacunas disponibles para su aplicación en humanos todavía tienen las desventajas asociadas con su reducida inmunogenicidad y la baja memoria inmunológica en las poblaciones de alto riesgo como los niños (i.e. vacunas de polisacáridos de neumococos) o con su elevado costo como estrategia de salud pública en comunidades de escasos recursos (i.e. vacunas conjugadas) . Por lo tanto, los actuales esfuerzos en el desarrollo de vacunas para neumococo se focalizan en la exploración de estrategias alternativas que permitan mejorar la eficacia y efectos de las formulaciones existentes. Con el objeto de mejorar la eficiencia de las vacunas, es necesario incluir nuevos tipos de adyuvantes, capaces de estimular a las células presentadoras de antígenos, induciendo la producción de citoquinas que favorezcan la presentación antigénica, la producción de anticuerpos específicos y la mejora de la memoria inmunológica. Entre estos tipos de adyuvantes, se encuentran componentes de origen bacteriano, capaces de estimular los receptores de reconocimiento de patrones (PRR), como por ejemplo el receptor de tipo Toll-2 (TLR2). Previamente, hemos demostrado que tanto Lactobacillus rhamnosus CRL1505 no viable como sus fracciones celulares (pared celular y péptidoglicano) tienen la capacidad de inducir una mejor respuesta inmune innata a nivel sistémico y respiratorio en animales desnutridos, al ser administrados por vía nasal u oral. Sin embargo, hasta el momento, no se ha evaluado la capacidad de sus fracciones celulares, empleadas como adyuvantes de vacunas de administración nasal, para conferir inmunidad protectora en poblaciones de riesgo. Por lo tanto, en este trabajo evaluamos si las fracciones celulares provenientes de la cepa inmunomoduladora L. rhamnosus CRL1505 pueden ser empleadas como adyuvantes de mucosa efectivos para la mejora de la eficiencia de la inmunización en ratones infantes. Más aún, estudiamos si la inmunización de ratones albino Suizos infantes con las fracciones celulares son capaces de mejorar la resistencia al patógeno respiratorio.Streptococcus pneumoniae is a major global health problem in high-riskgroups. Even when vaccines are available, they show reduced low immunological memory in these populations. Other disadvantageis the high cost as a public health strategy in communities with low incomes. Inthis context, immunobiotic bacteria such as Lactobacillus rhamnosus CRL1505 or its cellular fractions could represent candidates as mucosal adjuvant. Theaim of this work was to study the potential use of cell wall (PC) or peptidoglycan(PG) from the immunomodulatory strain L. rhamnosus CRL1505, aseffective mucosal adjuvant in infant and adult mice when are co-administered with the commercially available conjugated-polysaccharide 13-valent pneumococcal vaccine (Vc). Results showed that administration of the Vc induced production of pneumococcal-specific antibodies in serum and broncho-alveolarlavage (BAL) in adult and infant mice. Infant mice showed higher levels of specific IgM and IgA in serum, and specific IgA and IgG in BAL, respectto adults after nasal immunization. In addition, subcutaneous immunization induced higher antibody levels in adults than infant mice. Co-administrationof Vc+PC or Vc+PG induced higher pneumococcal-specific antibodies (IgM,IgA, IgG) compared to Vc alone in infant mice. To evaluate the capability of Vcand cell fractions to protect from infection, four S. pneumoniae serotypes (3,6B, 14, and 19F) were used. Only the nasal immunization with Vc+PC significantlyreduced the pneumococcal lung colonization by the 4 serotypes, whilethe combination Vc+PG reduced lung colonization by serotype 3 and 6B, withno effect on serotypes 14 and 19F. All treatments (Vc, Vc+PC, and Vc+PG) avoided pathogen dissemination into blood. The results suggest that cell wallfrom L. rhamnosus CRL1505 is an interesting and potential mucosal adjuvantto improve effectiveness of commercial vaccines and increasing protection in children.Fil: Laiño, Jonathan Emiliano. 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: Kolling, Yanina Noralí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Aguero Villoldo, Maria Graciela. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; ArgentinaFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentin

Copper deficiency in Creole goat kids

Serum copper determination is important to confirm hypocupremia. Twenty healthy kids constituted the Control Group, and sixteen kids with symptoms of copper deficiency, the Problem Group. Animals from the Problem Group showed a low number of red blood cells (RBC) and variations in RBC size and shape. The values found for hemoglobin and mean corpuscular hemoglobin (MCH) in the Control Group were 10.42 ± 1.34 g·dL-1 and 33.07 ± 1.11 g·dL-1 respectively, while the levels of the Problem Group were 7.95 ± 1.21 g·dL-1 and 29.45 ± 0.78 g·dL-1, respectively. The kids from the Problem Group presented an important increase in monocytes, neutrophiles and leukocytes; precursor cells of the neutrophile were also observed. The anemia of these animals was hypochromic and macrocytic. Our results indicate the Creole kids with serum copper levels > 450 μg·L-1 improved after treatment with copper glycinate. The six goats with cupremia < 450 μg·mL-1 were unable to improve their deficiency and died.Fil: Draksler, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Nuñez, Martha Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Apella, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; ArgentinaFil: Aguero Villoldo, Maria Graciela. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clínica Aplicada; ArgentinaFil: Gonzalez, Silvia Nelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentin

Immunomodulatory lactic acid bacteria in regulation of haemostatic alterations induced during a respiratory infection in immunosuppressed host by malnourishment. Basic mechanisms of action

Malnutrition induces a decrease in immunity that affects the ability of the organism to deal with an infectious challenge. The clotting system is considered a branch of immunity and its activation is important in the pathogenesis of an infectious disease. This work was conducted to determine coagulation modifications in malnourished hosts before and during infection. Weaned mice were malnourished via a protein-free diet. Well-nourished control mice (WNC) consumed a balanced conventional diet. Malnourished mice (MN) and WNC were challenged intranasally with Streptococcus pneumoniae. Blood, bronchoalveolar lavages (BAL), and lung samples were taken at different times post infection. The results were that MN showed altered hemostatic tests and fibrin(ogen) deposits in the lung. Thus, an increase in thrombin antithrombin complexes (TATc) in plasma and BAL was observed. In the MN group, infection induced a rise in TATc in plasma and BAL and increased plasma fibrinogen and fibrin(ogen) deposits in the lung. A decrease in activated protein C and antithrombin in BAL and an early decrease followed by an increase in plasma Factor VIII were also observed. Also MN demonstrated high levels of acute phase reactant and an increase in TNF-alfa. The feeding with a diet supplemented with Lactobacillus casei induced an anti-inflamatory cytokine pattern which contributed to limit coagulation activation and to normalize the regulatory protein. These findings will help to develop further strategies to reduce the damaging effects of clotting and enhance its beneficial contribution to immune reactions. Key words: malnourishment, blood coagulation, plasma, bronchoalveolar lavage, coagulation activation, coagulation inhibitors, TAT complexes.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; ArgentinaFil: Laiño, Jonathan Emiliano. 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: Haro, Ana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; ArgentinaFil: Aguero Villoldo, Maria Graciela. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; Argentin

Lactobacillus casei CRL431 modulates hemostatic activation induced by protein malnourishment and pneumococcal respiratory infection

Previously, we demonstrated that Lactobacillus casei CRL431, a well-known immunomodulatory bacterium, beneficially regulates coagulation activation, fibrin formation in lung, and the pro-inflammatory state induced by protein malnourishment and pneumococcal infection. In this study, we deepen in the understanding of the mechanisms involved in the immunoregulatory activity of L. casei CRL431 during a nutritional repletion process by evaluating (a) platelet and endothelial activation, (b) tissue factor (TF) expression, and (c) protease-activated receptor (PAR) activation in an experimental bacterial respiratory infection model in malnourished mice. Our findings demonstrate for the first time that the repletion diet supplemented with L. casei CRL431 was effective to normalize platelet counts in blood, modulate platelet activation and their recruitment into the lung, and regulate local and systemic TF expression and endothelial activation, which were affected by malnourishment. Streptococcus pneumoniae challenge induced local and systemic increase of platelet counts, PARs activation, P-selectin and TF expression, as well as endothelial activation in both well-nourished and malnourished mice. Malnourished animals evidenced the highest alterations of the parameters evaluated while the mice fed with the probiotic bacterium had similar behavior to normal controls but with lower PAR activation in lung. These results demonstrate that supplementation of repletion diet with L. casei CRL431 is effective to modulate alterations induced by malnourishment and pneumococcal infection, restraining coagulation activation, the inflammatory process, and lung damage. These observations contribute to set the basis for the application of probiotic functional foods to modulate the inflammation-hemostasis interactions altered by malnourishment or bacterial respiratory infections.Fil: Zelaya, María Hortensia del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Aplicada; ArgentinaFil: Laiño, Jonathan Emiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Marranzino, Gabriela. Universidad del Norte Santo Tomás de Aquino. Facultad de Ciencias de la Salud; ArgentinaFil: Alvarez, Gladis 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: Aguero Villoldo, Maria Graciela. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Clinica Aplicada. Cátedra de Bioquímica Clinica I; Argentin

Nasal priming with immunobiotic Lactobacillus rhamnosus modulates inflammation–coagulation interactions and reduces influenza virus-associated pulmonary damage

Objective: To evaluate the effect of the nasal administration of live and heat-killed Lactobacillus rhamnosus CRL1505 (Lr1505) on immune-coagulative response during influenza virus (IFV) infection to improve survival and reduce lung injury. Methods: Six-week-old BALB/c mice were treated with live or heat-killed Lr1505 by the nasal route during two consecutive days. Treated and untreated control mice were then nasally challenged with IFV. Results: Both viable and non-viable Lr1505 protected infected mice by reducing pulmonary injury and lung viral loads trough several mechanisms: (a) Inflammatory cytokines were efficiently regulated allowing higher clearance of virus and reduction of inflammatory lung tissue damage, associated to higher levels of the regulatory cytokine IL-10. (b) The antiviral immune response was enhanced with improved levels of type I interferons, CD4+IFN-g+ lymphocytes, and lung CD11c+CD11blow- CD103+ and CD11c+CD11bhighCD103- dendritic cells. (c) The procoagulant state was reversed mainly by downregulating tissue factor expression and restoring thrombomodulin levels in lung. The capacity of Lr1505 to improve the outcome of IFV infection would be related to its ability to beneficially modulate lung TLR3-triggered immune response. Conclusions: Our work is the first to demonstrate the ability of an immunobiotic strain to beneficially modulate inflammation-coagulation interactions during IFV infection. Interestingly, non-viable L. rhamnosus CRL1505 was as effective as the viable strain to beneficially modulate respiratory antiviral immune response.Fil: Zelaya, María Hortensia del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Aplicada; ArgentinaFil: Tada, Asuka. Tohoku University; JapónFil: Vizoso Pinto, María Guadalupe. 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; ArgentinaFil: Salva, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Kanmani, Paulraj. Tohoku University; JapónFil: Aguero Villoldo, Maria Graciela. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Aplicada; ArgentinaFil: Alvarez, Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Bioquímica Aplicada; 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; Argentina. Tohoku University; Japó