Limosilactobacillus fermentum CECT5716: Mechanisms and Therapeutic Insights

This work was supported by the Junta de Andalucía (CTS 164) and Instituto de Salud Carlos III (PI19/01058) with funds from the European Union.M.J. Rodríguez-Sojo is a predoctoral fellow from University of Granada (“Programa de Doctorado en Biomedicina”); A.J. Ruiz-Malagón is a predoctoral fellow from Formación de Profesorado Universitario Program (“Programa de Doctorado en Medicina Clínica y Salud Pública”), and A. Rodríguez-Nogales is a postdoctoral fellow of Instituto de Salud Carlos III (Miguel Servet Program).Probiotics microorganisms exert their health-associated activities through some of the following general actions: competitive exclusion, enhancement of intestinal barrier function, production of bacteriocins, improvement of altered microbiota, and modulation of the immune response. Among them, Limosilactobacillus fermentum CECT5716 has become one of the most promising probiotics and it has been described to possess potential beneficial effects on inflammatory processes and immunological alterations. Different studies, preclinical and clinical trials, have evidenced its anti-inflammatory and immunomodulatory properties and elucidated the precise mechanisms of action involved in its beneficial effects. Therefore, the aim of this review is to provide an updated overview of the effect on host health, mechanisms, and future therapeutic approaches.Junta de Andalucia CTS 164Instituto de Salud Carlos III European Commission PI19/01058European Commissio

Gut Microbiota Composition Can Predict Colonization by Multidrug-Resistant Bacteria in SARS-CoV-2 Patients in Intensive Care Unit: A Pilot Study

The SARS-CoV-2 infection has increased the number of patients entering Intensive Care Unit (ICU) facilities and antibiotic treatments. Concurrently, the multi-drug resistant bacteria (MDRB) colonization index has risen. Considering that most of these bacteria are derived from gut microbiota, the study of its composition is essential. Additionally, SARS-CoV-2 infection may promote gut dysbiosis, suggesting an effect on microbiota composition. This pilot study aims to determine bacteria biomarkers to predict MDRB colonization risk in SARS-CoV-2 patients in ICUs. Seventeen adult patients with an ICU stay >48 h and who tested positive for SARS-CoV-2 infection were enrolled in this study. Patients were assigned to two groups according to routine MDRB colonization surveillance: non-colonized and colonized. Stool samples were collected when entering ICUs, and microbiota composition was determined through Next Generation Sequencing techniques. Gut microbiota from colonized patients presented significantly lower bacterial diversity compared with non-colonized patients (p < 0.05). Microbiota in colonized subjects showed higher abundance of Anaerococcus, Dialister and Peptoniphilus, while higher levels of Enterococcus, Ochrobactrum and Staphylococcus were found in non-colonized ones. Moreover, LEfSe analysis suggests an initial detection of Dialister propionicifaciens as a biomarker of MDRB colonization risk. This pilot study shows that gut microbiota profile can become a predictor biomarker for MDRB colonization in SARS-CoV-2 patients.Junta de Andalucía (CTS 164; CV20-77708)Instituto de Salud Carlos III (PI19/01058; PI20/01447Fundación Andaluza de Farmacia Hospitalaria” (3095/2020)

Intestinal anti-inflammatory and visceral analgesic effects of a Serpylli herba extract in an experimental model of irritable bowel syndrome in rats

Ethnopharmacological relevance: Serpylli herba extract (SHE), composed of the aerial parts of wild thyme (Thymus serpyllum L.) (Lamiaceae family), is traditionally used in Europe and North Africa to treat diarrhea, gastric ulcers, intestinal parasites and upper respiratory tract infections. Recently, SHE has generated a great interest for irritable bowel syndrome (IBS) management, probably due to its intestinal anti-inflammatory properties shown in experimental colitis and the fact that its active components could preserve the intestinal barrier integrity, which is altered in patients with IBS. Aim of study: We aimed to test the effects of a SHE in a rat experimental model resembling human IBS. Materials and methods: IBS was provoked by deoxycholic acid (DCA). Rats were then treated with SHE (100 mg/kg) or gabapentin (70 mg/kg) and different inflammatory and gut barrier integrity markers were evaluated. Moreover, several gut hypersensitivity and hyperalgesia determinations were performed. Results: SHE improved referred pain and visceral hypersensitivity. Additionally, SHE enhanced immune status by downregulating of the expression of the proinflammatory mediators Il-1β, Il-6, Ifn-γ, Tlr-4, and the inducible enzyme Cox-2, thus inducing visceral analgesia, and promoting the restore of the gut barrier function by upregulating the mucins Muc-2 and Muc-3. These antiinflammatory effects could be related to its action on mast cells since it significantly inhibited the β-Hexosaminidase production in RBL-2H3 cells. Lastly, SHE also seems to modulate the serotonin pathway by restoring the altered expression of the 5-HT receptors Htr-3 and Htr-4. Conclusion: SHE could be considered a potential new treatment for IBS, since it ameliorates hypersensitivity, visceral hyperalgesia, and inflammation. These beneficial effects may be due to the inhibition of mast cells degranulation and serotonin pathway.Junta de Andalucia AGR6826 CTS 164Spanish Government SAF 2011-29648Instituto de Salud Carlos IIIEuropean Commission PI19/01058 PI20/0144

Novel Horizons in Postbiotics: Lactobacillaceae Extracellular Vesicles and Their Applications in Health and Disease

Lactobacillus probiotics contained in dietary supplements or functional foods are wellknown for their beneficial properties exerted on host health and diverse pathological situations. Their capacity to improve inflammatory bowel disease (IBD) and regulate the immune system is especially remarkable. Although bacteria–host interactions have been thought to occur directly, the key role that extracellular vesicles (EVs) derived from probiotics play on this point is being unveiled. EVs are lipid bilayer-enclosed particles that carry a wide range of cargo compounds and act in different signalling pathways. Notably, these EVs have been recently proposed as a safe alternative to the utilisation of live bacteria since they can avoid the possible risks that probiotics may entail in vulnerable cases such as immunocompromised patients. Therefore, this review aims to give an updated overview of the existing knowledge about EVs from different Lactobacillus strains, their mechanisms and effects in host health and different pathological conditions. All of the information collected suggests that EVs could be considered as potential tools for the development of future novel therapeutic approaches.Junta de Andalucia CTS 164 PY20_01157Instituto de Salud Carlos III European Commission PI19/01058 European Commissio

Anti-Inflammatory and Chemopreventive Effects of Bryophyllum pinnatum (Lamarck) Leaf Extract in Experimental Colitis Models in Rodents

Inflammatory bowel diseases, mainly ulcerative colitis and Crohn’s disease are characterized by chronic inflammation in the intestine. Currently several therapeutic strategies available to treat inflammatory bowel diseases. Though, most treatments can be associated with serious adverse effects what justifies the search for new treatments. In this sense, we highlight the interest in herbal products rich in bioactive compounds which immunomodulatory and antioxidant properties as is the case of Bryophyllum pinnatum (Crassulaceae). This plant is used in traditional medicine in Brazil for treating inflammatory diseases. We hypothesized that hydroethanolic B. pinnatum leaf extract has intestinal anti-inflammatory effects on two experimental colitis models: 2.4-dinitrobenzene sulfonic acid (DNBS) in rats, and dextran sulfate sodium (DSS) in mice. Ultra-fast liquid chromatography method used for the quantification of the main compounds indicated good linearity, specificity, selectivity, precision, robustness and accuracy. The major flavonoids (mg/g of the extract) quantified were: quercetin 3-O-a-L-arabinopyranosyl-(1!2)-a-L-rhamnopyranoside (35.56 ± 0.086 mg/g), kaempferol 3-O-a-L-arabinopyranosyl-(1!2)-a-L-rhamnopyranoside (4.66 ± 0.076 mg/g) and quercetin-3-O-rhamnopyranoside (4.56 ± 0.026 mg/g). The results obtained in the DNBS and DSS models indicate that extract has both chemopreventive and antiinflammatory effects, observing a significant reduction in the disease activity index score, and less macroscopic and microscopic damage. The extract promoted downregulation of Tolllike receptor and kappa B p65 nuclear factor gene expression, leading to a reduction in pro-inflammatory and oxidative mediators, chemokines, and cell adhesion molecules. This immunomodulatory property was proposed that one of the possible action mechanisms of extract. An improvement in intestinal damage was also associated with a reduction in oxidative stress and infiltration of leukocytes, as evidenced by the reduction in malonaldialdehyde and myeloperoxidase activity and increase in total glutathione in the colonic tissue. Moreover, the extract improved the cytoarchitecture of the colonic tissue and the integrity of the intestinal epithelial barrier by restoring the expression of the proteins associated with mucosa protection. In view of the beneficial effects showed by the B. pinnatum leaf extract in preclinical rodent models of colitis there is the potential to conduct some future clinical studies to ensure safe and effective development of a phytotherapeutic treatment for human inflammatory bowel diseases.CAPESJunta de Andalucia CTS 164Spanish Ministry of Economy and Competitiveness AGL2015-67995-C3-3-REuropean Union (EU)Instituto de Salud Carlos II

Calcium Pyruvate Exerts Beneficial Effects in an Experimental Model of Irritable Bowel Disease Induced by DCA in Rats

Pyruvate is a normal constituent of the body that participates in carbohydrate metabolism and functions as a scavenger of free radicals. Calcium pyruvate monohydrate (CPM) is a more stable derivative that has proved its anti-inflammatory effect in experimental colitis, among other disorders, and that could also be considered a source of calcium. Thus, it would be useful for the treatment of diseases with an inflammatory component and a high prevalence of osteoporosis like the irritable bowel syndrome (IBS). The aim of the present study is to evaluate the effects of CPM in a rat model of chronic post-inflammatory visceral pain induced by deoxycholic acid (DCA) that resembles IBS. Rats were administered DCA for three days intracolonically and then treated daily with CPM (40 and 100 mg/kg) or gabapentin (70 mg/kg) (positive control) by oral gavage for 17 days. The treatments reduced the visceral hypersensitivity measured by response to colorectal distension and referred pain. DCA induced changes in the colonic immune response characterized by increased expression of the cytokine Il-1b and the inducible enzyme Cox-2, which was reduced by the treatments. DCA also decreased the gut expression of the mucins Muc-2 and Muc-3, which was normalized by CPM, whereas gabapentin only increased significantly Muc-3. Moreover, DCA increased the expression of Tlr3, which was decreased to basal levels by all the treatments. However, the serotonin receptor Htr-4, which was also elevated, was not affected by any of the treatments, indicating no effect through this signalling pathway. In conclusion, CPM ameliorated the visceral hypersensitivity and the referred pain caused by DCA, being as effective as the control drug. Furthermore, it improved the immune status of the animals, which could contribute to the visceral analgesia and the regeneration of the intestinal epithelial barrier integrity.This work was supported by the Junta de Andalucía (CTS 164) and by the Spanish Ministry of Economy and Competitiveness (AGL2015-67995-C3-3-R) with funds from the European Union. The CIBER-EHD is funded by the Instituto de Salud Carlos III

Tigecycline reduces tumorigenesis in colorectal cancer via inhibition of cell proliferation and modulation of immune response

Junta de Andalucía (CTS 164)Instituto de Salud Carlos III (Spain)Fondo Europeo de Desarrollo Regional (FEDER)European Union, through the research grants PI18/00826, P18-RT-4930, PI0206–2016, PIE16/00045 and PI19/01058Spanish Ministry of Science and Innovation (MCIN/AEI/ 10.13039/501100011033/FEDER)RTI2018–101309-BC22Chair “Doctors Galera-Requena in cancer stem cell research” (CMC-CTS963Spanish Ministry of Science and Innovation (“Programa de Doctorado: Medicina Clínica y Salud Pública” B12.56.1).Instituto de Salud Carlos III (FI17/00176)Junta de Andalucía (P18-RT-4930)University of GranadaCIBER-EHD is funded by the Instituto de Salud Carlos II

Beneficial Effects of Limosilactobacillus fermentum in the DCA Experimental Model of Irritable Bowel Syndrome in Rats

Limosilactobacillus fermentum CECT5716, a probiotic strain isolated from human milk, has reported beneficial effects on different gastrointestinal disorders. Moreover, it has shown its ability to restore altered immune responses, in association with microbiome modulation in different pathological conditions. Therefore, our aim was to assess the effects of a Limosilacbacillus fermentum CECT5716 in a rat experimental model of irritable bowel syndrome (IBS) that resembles human IBS. The experimental IBS was induced by deoxycholic acid (DCA) in rats and then, Limosilactobacillus fermentum CECT5716 (109 CFU/day/rat) was administered. Behavioral studies, hyperalgesia and intestinal hypersensitivity determinations were performed and the impact of the probiotic on the inflammatory and intestinal barrier integrity was evaluated. Additionally, the gut microbiota composition was analyzed. Limosilactobacillus fermentum CECT5716 attenuated the anxiety-like behavior as well as the visceral hypersensitivity and referred pain. Moreover, this probiotic ameliorated the gut inflammatory status, re-establishing the altered intestinal permeability, reducing the mast cell degranulation and re-establishing the gut dysbiosis in experimental IBS. Therefore, our results suggest a potential use of Limosilactobacillus fermentum CECT5716 in clinical practice for the management of IBS patients.Junta de Andalucia A-CTS-447-UGR18 CTS 164Instituto de Salud Carlos IIIEuropean Commission PI19/01058 PI20/0144

Macrofauna asociada al alga stypocaulon scoparium en el Estrecho de Gibraltar y comparación con el resto de la Península Ibérica

Se estudió la macrofauna asociada al alga Stypocaulon scoparium en el estrecho de Gibraltar y el resto de la península Ibérica. Se seleccionaron un total de 14 estaciones, 3 de ellas localizadas en el Estrecho y las 11 restantes distribuidas en las costas cantábricas, atlánticas y mediterráneas de la península Ibérica. En cada estación se midieron parámetros fisicoquímicos (temperatura, oxígeno disuelto, conductividad, pH y turbidez), se estimó la cobertura del alga y se recolectaron muestras de la misma en el submareal somero (1- 3 m de profundidad) mediante buceo en apnea. Se separaron e identificaron un total de 48.430 individuos pertenecientes a 16 grupos distintos de artrópodos, moluscos, anélidos y equinodermos. Atendiendo a los parámetros fisicoquímicos, los análisis de clasificación mostraron una mayor similaridad del área del Estrecho con las estaciones atlánticas. La costa mediterránea se caracterizó por una mayor temperatura y conductividad, mientras que la atlántica mostró valores más altos de oxígeno disuelto y turbidez, consecuencia un mayor oleaje. La cobertura del alga S. scoparium fue significativamente mayor en el Estrecho de Gibraltar que en las estaciones restantes y se registró una mayor diversidad de invertebrados asociados a este alga en el Estrecho que en otras zonas de la Península. Los crustáceos anfípodos fueron los más abundantes, seguidos de poliquetos y moluscos gasterópodos. El Análisis Canónico de Correspondencias (CCA) mostró que anfípodos, isópodos, quironómidos y crinoideos correlacionaron principalmente con la cobertura del alga, mientras que la abundancia de gasterópodos estuvo condicionada por la turbidez y la de los grupos restantes por la conductividad y temperatura principalmente. El estrecho d

Patrones de variación espacial de las comunidades intermareales de la Isla de Tarifa (Cádiz)

El ecosistema mediolitoral se encuentra muy deteriorado en la mayor parte de la costa andaluza debido a la facilidad de acceso y la consiguiente presión humana. Sin embargo, la Isla de Tarifa refleja un estado de excelente conservación, en gran medida consecuencia de las restricciones de acceso y de su localización biogeográfica. Se seleccionaron un total de cinco estaciones distribuidas a lo largo de todo el perímetro de la Isla y se estudió la distribución vertical de las principales especies de macroalgas y de fauna sésil utilizándose cuadrículas de 50x50cm subdivididas en 25 subcuadrículas. Los valores de temperatura, salinidad, pH y concentración de oxígeno fueron similares en todas las estaciones. Las mayor diversidad se registró en los cinturones intermedios. Las especies de algas dominantes en todas las estaciones muestreadas fueron Asparagopsis armata y Corallina elongata que ocuparon los niveles más bajos, próximos al submareal, y el césped de Caulacanthus ustulatus y Gelidium spp que se extendió en los cinturones intermedios. En cuanto a la fauna sésil, estuvo dominada por el crustáceo cirrípedo Chatamalus stellatus y el molusco Littorina neritoides que ocuparon los niveles más altos. Las lapas Patella caerulea y P. rustica también fueron abundantes en la franja intermareal; la primera se distribuyó en la zona intermedia mientras que la segunda estuvo restringida a los niveles superiores. Las algas Codium adherens y Colpomenia sinuosa mostraron preferencia por las estaciones con mayor influencia del sedimento; Ulva rigida, Valonia utricularis y Osmundea pinnatifida predominaron en las estaciones expuestas a vientos de levante, mientras que Chaetomorpha linum se encontró en los cinturones expuestos al poniente. En cuanto a la fauna, el percebe Pollicipes pollicipes restringió su presencia a las estaciones de mayor grado de exposición, el pulmonado Siphonaria pectinata se registró en las estaciones más embolsadas y el cirrípedo C. stellatus mostró una clara preferencia por zonas expuestas a vientos de poniente. En el intermareal de la Isla de Tarifa se distribuyen las especies amenazadas Astroides calycularis, Dendropoma petraeum, Patella ferruginea y Cymbula nigra, lo que debe incrementar su valor en las estrategias de conservación.The intertidal ecosystems are seriously damaged in most of the Andalousian coast, mainly due to accessibility and human effect. However, Tarifa Island, is still in an excellent state of conservation, due to the access restrictions and unique biogeographical position. Five stations were selected along the island and vertical distribution of the main species of macroalgae and sessile fauna were studies using quadrats of 50x50cm (subdivided in 25 square units). Temperature, salinity, pH and oxygen concentration were similar in all the stations. The highest diversity was measured in the intermediate belts. The dominant species of algae were Asparagopsis armata and Corallina elongata, distributed in the lowest levels close to the subtidal, and the turf of Caulacanthus ustulatus and Gelidium spp, which occupied the intermediate levels. Regarding with the fauna, the crustacean Chtamalus stellatus and the mollusc Littorina neritoides dominated the highest levels. The limpets Patella caerulea and P. rustica were also abundant, the former distributed in the intermediate zone and the latter restricted to the upper belts. The algae Codium adherens and Colpomenia sinuosa preferred stations with sediment influence, Ulva rigida, Valonia utricularis and Osmundea pinnatifida were dominant in the stations exposed to east winds, whereas Chaetomorpha linum was found in sites exposed to west winds. In connection with the fauna, the barnacle Pollicipes pollicipes was only present in the most exposed stations, Siphonaria pectinata was registered in the less exposed sites and C. stellatus dominated the stations exposed to west winds. The endangered species Astroides calycularis, Dendropoma petraeum, Patella ferruginea and Cymbula nigra are also present along Tarifa intertidal, which should increase the value of the site in conservation policies.Ministerio de Educación y Ciencia CGL2007-60044/BOSConsejería de Innovación, Ciencia y Empresa de la Junta de Andalucía P07-RNM- 0252