Effect of in-ovo feeding of iron nanoparticles and methionine hydroxy analogue on broilers chickens small intestinal characteristics

The experiment was conducted with 644 Ross fertilized egg by 7 treatments 4 replicates and 23 eggs in each. Seven treatments included two control with and without injection, iron sulfate, iron sulfate nanoparticles, Alimet, Alimet + iron sulfate, Alimet + iron sulfate nanoparticles. After hatching 2 mg iron nanoparticles were applied as new treatment. The highest increased in the intestinal relative weight (p < 0.05) was observed by iron+Alimet in late feeding at day old of age. Also similar trend was found in cecum and duodenum length by iron control 2 and late feeding (18 hours after hatching). The highest cecum length was found among all treatments by in ovo injection of iron nanoparticles in early feeding at 21 days of age (p < 0.05). Significantly increased the duodenum length was found by iron sulfate in early feeding at 42 days of age (p < 0.05). In ovo injection of Alimet in late feeding was resulted in decrease jejunum crypt depth at 21 days of age (p < 0.05). The results of this study have shown that the highest jejunum villi width and surface area were recorded in dietary iron sulfate nanoparticles in late feeding at 21 and 42 days of age (p < 0.05).(AU)

Effect of in-ovo feeding of iron nanoparticles and methionine hydroxy analogue on broilers chickens small intestinal characteristics

The experiment was conducted with 644 Ross fertilized egg by 7 treatments 4 replicates and 23 eggs in each. Seven treatments included two control with and without injection, iron sulfate, iron sulfate nanoparticles, Alimet, Alimet + iron sulfate, Alimet + iron sulfate nanoparticles. After hatching 2 mg iron nanoparticles were applied as new treatment. The highest increased in the intestinal relative weight (p < 0.05) was observed by iron+Alimet in late feeding at day old of age. Also similar trend was found in cecum and duodenum length by iron control 2 and late feeding (18 hours’ after hatching). The highest cecum length was found among all treatments by in ovo injection of iron nanoparticles in early feeding at 21 days of age (p < 0.05). Significantly increased the duodenum length was found by iron sulfate in early feeding at 42 days of age (p < 0.05). In ovo injection of Alimet in late feeding was resulted in decrease jejunum crypt depth at 21 days of age (p < 0.05). The results of this study have shown that the highest jejunum villi width and surface area were recorded in dietary iron sulfate nanoparticles in late feeding at 21 and 42 days of age (p < 0.05).

Evaluation of Comparative Effect of Feed Additive of Allium Sativum and Zingeber Officinale on Bird Growth and Histomorphometric Characteristics of Small Intestine in Broilers

The present study was conducted to evaluate the effects of supplementation of garlic, ginger in the diets of broiler chickens and assessment in terms of feed intake, growth performance and economics of feeding. The results showed that groups supplemented with 0.5% garlic powder and 0.5% ginger powder has shown significant effects on body weight as compared to the control group at day 28. Groups supplemented with 0.5% garlic powder and 0.5% ginger powder show significant increase in body weight than the groups supplemented with 0.25% garlic powder and 0.25% ginger powder. Between different supplemented groups, villus length and width of duodenum and jejunum of birds served with 0.5% garlic powder and 0.5% ginger powder is significantly higher than the villus length and width of birds supplemented with 0.25% garlic and 0.25% ginger powder. Between different supplemented groups, villus length of ileum of the group supplemented with 0.5% garlic powder is significantly (p 0.05) lower than the villus length of the groups supplemented with 0.25% garlic powder and 0.25% ginger powder.

Evaluation of Comparative Effect of Feed Additive of Allium Sativum and Zingeber Officinale on Bird Growth and Histomorphometric Characteristics of Small Intestine in Broilers

The present study was conducted to evaluate the effects of supplementation of garlic, ginger in the diets of broiler chickens and assessment in terms of feed intake, growth performance and economics of feeding. The results showed that groups supplemented with 0.5% garlic powder and 0.5% ginger powder has shown significant effects on body weight as compared to the control group at day 28. Groups supplemented with 0.5% garlic powder and 0.5% ginger powder show significant increase in body weight than the groups supplemented with 0.25% garlic powder and 0.25% ginger powder. Between different supplemented groups, villus length and width of duodenum and jejunum of birds served with 0.5% garlic powder and 0.5% ginger powder is significantly higher than the villus length and width of birds supplemented with 0.25% garlic and 0.25% ginger powder. Between different supplemented groups, villus length of ileum of the group supplemented with 0.5% garlic powder is significantly (p 0.05) lower than the villus length of the groups supplemented with 0.25% garlic powder and 0.25% ginger powder.(AU)

Long-Term Dietary Intake of Chia Seed Is Associated with Increased Bone Mineral Content and Improved Hepatic and Intestinal Morphology in Sprague-Dawley Rats.

Chia seeds (Salvia hispanica) provide an unusually high content of α-linolenic acid with several potential health benefits, but few studies have examined the long-term intake of n-3 fatty acid-rich plant foods such as chia. In this work, we investigated some of the effects of a diet containing 10% chia seeds versus a conventional isocaloric diet for 10 and 13 months on body measurements, musculoskeletal system, the liver, and the intestines of 20 male Sprague-Dawley rats assigned into two groups. The n-6/n-3 ratios for the control and chia diets were 7.46 and 1.07, respectively. For the first 10 months of the diet, the body parameters and weights were similar, but at 13 months, the bone mineral content (BMC) of the chia-fed rats was significantly higher than that of the controls whether in total or proximal areas of the left tibia. Also, significant positive correlations were found between the age of the chia group and the bone mineral density, BMC, weight of the musculoskeletal system, final body weight, and skin weight. Liver and intestinal examinations showed improved morphology associated with lower lipid deposit in hepatocytes and increased intestinal muscle layers and crypt size in the chia group. This study provides new data suggesting the potential benefits associated with the long-term intake of chia seeds.

Comparison of Human Denuded Amniotic Membrane and Porcine Small Intestine Submucosa as Scaffolds for Limbal Mesenchymal Stem Cells.

Blinding corneal scarring is usually treated with allogeneic graft tissue. Nevertheless, the global shortage of donors leaves millions of patients in need of therapy. Traditional tissue engineering strategies involves the combination of cells, growth factors, and scaffolds that can supply cellular biological components allowing to restore the tissue function. The mesenchymal stem cells found in the limbal stroma (L-MSCs) have a self-renewal potential for multilineage differentiation. Thus, in this work we compared the potential of human amniotic membrane (hAM) and porcine small intestine submucosa (SIS) as scaffolds for L-MSCs, aiming at potential applications in corneal regeneration. For that, L-MSCs were seeded on hAM and SIS and we analyzed their viability, actin cytoskeleton, nuclei morphology, cell density, adhesion and surface markers. Our results showed that cells adhered and integrated into both membranes with a high cell density, an important characteristic for cell therapy. However, due to its transparency, the hAM allowed a better observation of L-MSCs. In addition, the analysis of surface markers expression on L-MSCs after two weeks showed a slight increase in the percentages of negative markers for MSCs grown on SIS membrane. Thus, considering a long-term culture, the hAM was considered better in maintaining the MSCs phenotype. Regarding the function as scaffolds, SIS was as efficient as the amniotic membrane, considering that these two types of biological matrices maintained the cell viability, actin cytoskeleton, nuclei morphology and mesenchymal phenotype, without causing cell death. Therefore, our data in vitro provides evidence for future pre-clinical studies were these membranes can be used as a support to transport mesenchymal stem cells to the injured area, creating a kind of temporary curative, allowing the release of bioactive molecules, such as cytokines and growth factors and then promoting the tissue regeneration, both in human and veterinary medicine.

Recreating Intestinal Peristalsis in the Petri Dish.

Here we describe a culture technique of cells dissociated from the external muscularis of the guinea pig small intestine, which allows us to maintain all the elements involved in the intestinal peristaltic reflex. After a few days in culture, these cells reorganize to form a small group of cells that permit the generation of pacemaker activity, spontaneous contractions, and the development of inhibitory and excitatory junction potentials in the petri dish, all elements involved in the peristaltic reflex. Therefore, these co-cultures are suitable to study the cellular and molecular aspects related to the development, maintenance, and modulation of motor intestinal functions.

Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase.

The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1ß, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology.

Mechanotransduction in small intestinal submucosa scaffolds: fabrication parameters potentially modulate the shear-induced expression of PECAM-1 and eNOS.

In small intestinal submucosa (SIS) scaffolds for functional tissue engineering, the impact of scaffold fabrication parameters on cellular response and tissue regeneration may relate to the mechanotransductory properties of the final arrangement of collagen fibres. We previously proved that two fabrication parameters, (a) preservation (P) or removal (R) of a dense collagen layer present in SIS, and (b) SIS in a final dehydrated (D) or hydrated (H) state, have an effect on the micromechanical environment of SIS. In a continuation of our studies, we herein hypothesized that these fabrication parameters also modulate early mechanotransduction in cells populating the scaffold. Mechanotransduction was investigated by seeding human umbilical vein endothelial cells (HUVECs) on scaffolds, exposing them to pulsatile shear stress (12 ± 4 dyne/cm2 ) for 1 h (n = 5) in a cone-and-plate shear system, and evaluating the expression of the mechanosensitive genes Pecam1 and Enos by immunofluorescence and qPCR. Expression of mechanosensitive genes was highest in PD grafts, followed by PH and RH grafts. The RD group had similar expression to that of unsheared control cells, suggesting that the RD combination potentially reduced mechanotransduction of shear to cells. We concluded that the two fabrication parameters studied, which modify SIS micromechanics, also potentially modulated the early shear-induced expression of mechanosensitive genes in seeded HUVECs. Our findings suggest that fabrication parameters influence the outcome of SIS as a therapeutic scaffold. Copyright © 2015 John Wiley & Sons, Ltd.

Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase

The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology.

Neuro-immune Interactions Drive Tissue Programming in Intestinal Macrophages.

Proper adaptation to environmental perturbations is essential for tissue homeostasis. In the intestine, diverse environmental cues can be sensed by immune cells, which must balance resistance to microorganisms with tolerance, avoiding excess tissue damage. By applying imaging and transcriptional profiling tools, we interrogated how distinct microenvironments in the gut regulate resident macrophages. We discovered that macrophages exhibit a high degree of gene-expression specialization dependent on their proximity to the gut lumen. Lamina propria macrophages (LpMs) preferentially expressed a pro-inflammatory phenotype when compared to muscularis macrophages (MMs), which displayed a tissue-protective phenotype. Upon luminal bacterial infection, MMs further enhanced tissue-protective programs, and this was attributed to swift activation of extrinsic sympathetic neurons innervating the gut muscularis and norepinephrine signaling to ß2 adrenergic receptors on MMs. Our results reveal unique intra-tissue macrophage specialization and identify neuro-immune communication between enteric neurons and macrophages that induces rapid tissue-protective responses to distal perturbations.

Morphological and Immunohistochemical Identification of Villous M Cells in the Small Intestine of Newborn Piglets / Identificación Morfológica e Inmunohistoquímica de los Epiteliocitos Microplegados de las Vellosidades en el Intestino Delgado de Lechones Recién Nacidos

Microfold (M) cells act as antigen-sampling sites for initiating antigen specific mucosal immune responses, but they may also provide a gateway for enteropathogen entry. In this study we demonstrated villous M cells by morphological and immunohistochemical methods to be present in the three regions of the small intestine from newborn piglets. Immunohistochemical analysis, using anti- cytokeratin 18 (CK18) primary antibodies, showed a gradually decreased density of M cells from the lower crypt epithelium to the upper villous epithelium. The proportion of villous M cells was greater in the ileum than in the duodenum or the mid-jejunum. Ultrastructural observation revealed that villous M cells were mainly columnar in shape in the duodenum and the mid-jejunum, and appeared as more pocket-like structure in the ileum. These villous M cells exhibited short and irregular microvilli, rich vesicles and reduced glycocalyx, but lacked the lymphocyte-containing basolateral invagination. Our results support evidence that M cells can develop in the small intestinal villous epithelium of newborn piglets, implying that villous M cells may begin developing in the pig's small intestine during fetal stages, which depends neither on the influence of the mucosal lymphoid tissue nor the antigen from the intestinal lumen stimulation. In addition, the variable morphology and heterogeneity distribution of villous M cells in the three regions of the small intestine may be indicative of its different functional properties. This information extent our understanding of the diversity of M cells and provides important basic knowledge for further research on the actual role of villous M cells in neonate.

Los epiteliocitos microplegados (células M) actúan como receptores de antígeno para iniciar la respuesta inmune específica de las mucosas, pero también pueden proporcionar una puerta de entrada para enteropatógenos. En este estudio, se demostró por métodos morfológicos e inmunohistoquímicos que los epiteliocitos microplegados de las vellosidades están presentes en las tres regiones del intestino delgado de lechones recién nacidos. Se utilizaron anticuerpos primarios de citoqueratina 18 (CK18) para el análisis inmunohistoquímico, el cual mostró una disminución gradual de la densidad de los epiteliocitos microplegados desde el epitelio de las criptas inferiores hasta el epitelio de las vellosidades superiores. La proporción de los epiteliocitos microplegados, fue mayor en el íleon que el duodeno o yeyuno medio. La observación ultraestructural reveló que los epiteliocitos microplegados fueron principalmente de forma columnar en el duodeno y el yeyuno medio. Además, mostraron microvellosidades cortas e irregulares, muchas vesículas y glucocáliz reducidos, pero carecían de invaginaciones basolaterales contenedoras de linfocitos. Nuestros resultados apoyan la evidencia de que los epiteliocitos microplegados pueden desarrollarse en el epitelio de las vellosidades intestinales de los lechones recién nacidos, lo que implica que estas células pueden comenzar a desarrollarse en el intestino delgado del cerdo durante las etapas fetales, y no dependen ni de la influencia del tejido linfoide de las mucosas ni del antígeno para la estimulación del lumen intestinal. Además, la morfología y heterogeneidad de distribución de los epiteliocitos microplegados en las tres regiones del intestino delgado pueden ser indicativas de sus diferentes propiedades funcionales. Esta información mejora nuestra comprensión de la diversidad de los epiteliocitos microplegados y proporciona conocimientos básicos importantes para la investigación sobre el papel de los epiteliocitos microplegados en las vellosidades del neonato.

Cdc42 coordinates proliferation, polarity, migration, and differentiation of small intestinal epithelial cells in mice.

BACKGROUND & AIMS: Cdc42 is a Rho GTPase that regulates diverse cellular functions, including proliferation, differentiation, migration, and polarity. In the intestinal epithelium, a balance among these events maintains homeostasis. We used genetic techniques to investigate the role of Cdc42 in intestinal homeostasis and its mechanisms. METHODS: We disrupted Cdc42 specifically in intestinal epithelial cells by creating Cdc42flox/flox-villin-Cre+ and Cdc42flox/flox-Rosa26-CreER+ mice. We collected intestinal and other tissues, and analyzed their cellular, molecular, morphologic, and physiologic features, compared with the respective heterozygous mice. RESULTS: In all mutant mice studied, the intestinal epithelium had gross hyperplasia, crypt enlargement, microvilli inclusion, and abnormal epithelial permeability. Cdc42 deficiency resulted in defective Paneth cell differentiation and localization without affecting the differentiation of other cell lineages. In mutant intestinal crypts, proliferating stem and progenitor cells increased, compared with control mice, resulting in increased crypt depth. Cdc42 deficiency increased migration of stem and progenitor cells along the villi, caused a mild defect in the apical junction orientation, and impaired intestinal epithelium polarity, which can contribute to the observed defective intestinal permeability. The intestinal epithelium of the Cdc42flox/flox-villin-Cre+ and Cdc42flox/flox-Rosa26-CreER+ mice appeared similar to that of patients with microvillus inclusion disease. In the digestive track, loss of Cdc42 also resulted in crypt hyperplasia in the colon, but not the stomach. CONCLUSIONS: Cdc42 regulates proliferation, polarity, migration, and differentiation of intestinal epithelial cells in mice and maintains intestine epithelial barrier and homeostasis. Defects in Cdc42 signaling could be associated with microvillus inclusion disease.

Endocrine cells and nerve ganglia of the small intestine of the Opossum Didelphis aurita Wied-Neuwied, 1826 (Mammalia: Didelphidae).

The nervous and endocrine systems jointly control intestinal movements, secretions of their glands and also participate of the processes of nutrient digestion and absorption. Therefore, the central objective of this study was to verify the existence of a possible relationship between the number of nervous cells and ganglia of the submucosal and myenteric plexuses and the number of endocrine cells in the small intestine of adult D. aurita. The utilized staining techniques were Grimelius, modified Masson-Fontana, direct immunoperoxidase and H-E. Argyrophillic, argentaffin and insulin immunoreactive endocrine cells do not numerically vary between the initial, mid and final regions of the duodenum, jejunum and ileum (P>0.05), except for argyrophillic cells in the jejunum (P>0.05). No numerical relationship has yet been verified between the number of nerve ganglia and endocrine cells, and also between nervous and endocrine cells. We recommended the use of new immunohistochemical techniques to confirm the numerical correlation between the nervous and endocrine systems in the small intestine. The morphology and distribution of endocrine cells and the nerve ganglia studied were similar to those encountered in eutherian mammals.

Characterizing the presence and sensitivity of the P2X7 receptor in different compartments of the gut.

Purinergic signaling has been established as an important feature of inflammation and homeostasis. The expression of a number of P2 receptor subtypes in the gut has been reported. In this study, using a well-known permeabilization method that is assessed by flow cytometry, we show that lymphocytes and macrophages from the mesenteric lymph nodes (MLN) and the peritoneal cavity exhibit different sensitivities to extracellular ATP. Compared with the macrophages, the lymphocytes are more sensitive to ATP in the MLN compartment, whereas in the peritoneal cavity the macrophages are more sensitive to ATP than the lymphocytes. In addition, we have shown that the epithelial cells from the small bowel are more resistant to the ATP effects than the cells from the colon. These cells, however, become susceptible after exposure to IFN-γ. Furthermore, by examining parameters such as pH manipulation, the exposure to divalent cations and the P2X7 antagonist Brilliant Blue G, and the use of cells from P2X7(-/-) mice, we have shown that the P2X7 receptors are the ATP-activated receptors responsible for the permeabilization phenomenon. In addition, using Western blot analysis, we have demonstrated the changes in the P2X7 receptor expression in immune cells isolated from different sites in the gut and in the gut-associated lymphoid tissues. Our findings suggest the existence of the site-specific modulation of P2X7 receptors on epithelial and immune cells, and we define purinergic signaling as a new regulatory element in the control of inflammation and cell fate in the gut and in the gut-associated lymphoid tissues.

Phenotypical and functional analysis of intraepithelial lymphocytes from small intestine of mice in oral tolerance.

In this work, we evaluated the effects of administration of OVA on phenotype and function of intraepithelial lymphocytes (IELs) from small intestine of transgenic (TGN) DO11.10 and wild-type BALB/c mice. While the small intestines from BALB/c presented a well preserved structure, those from TGN showed an inflamed aspect. The ingestion of OVA induced a reduction in the number of IELs in small intestines of TGN, but it did not change the frequencies of CD8(+) and CD4(+) T-cell subsets. Administration of OVA via oral + ip increased the frequency of CD103(+) cells in CD4(+) T-cell subset in IELs of both BALB/c and TGN mice and elevated its expression in CD8ß(+) T-cell subset in IELs of TGN. The frequency of Foxp3(+) cells increased in all subsets in IELs of BALB/c treated with OVA; in IELs of TGN, it increased only in CD25(+) subset. IELs from BALB/c tolerant mice had lower expression of all cytokines studied, whereas those from TGN showed high expression of inflammatory cytokines, especially of IFN-γ, TGF-ß, and TNF-α. Overall, our results suggest that the inability of TGN to become tolerant may be related to disorganization and altered proportions of inflammatory/regulatory T cells in its intestinal mucosa.

[Interaction of rotavirus with protein disulfide isomerase in vitro and cell system]. / Interacción de rotavirus con la proteína disulfuro-isomerasa in vitro y en sistemas celulares.

INTRODUCTION: Rotavirus entry process involves a multi-step mechanism, the first of which is when the outermost viral proteins interact with four different integrins and Hsc70. Recently, rotavirus infection reportedly has been decreased after blocking cell surface protein disulfide isomerase (PDI). This suggested that this protein interacts with rotavirus during the entry process. OBJECTIVES: The aim was to establish the rotavirus-PDI interaction in an in vitro system using PDI isolated from bovine liver, and in a cell system consisting of MA104 cells and mouse small intestinal villi. MATERIALS AND METHODS: Protein disulfide isomerase was isolated from a bovine liver homogenate using anti-PDI antibodies coupled to agarose through hydrazone bonds. Purity of purified protein was assessed by SDS-PAGE and Western blot. The purified PDI was used to study its in vitro interaction with the rotavirus particles. This interaction was compared with that taking place in MA104 cells and small intestinal villi isolated from sucking mice ICR. RESULTS: The purified PDI showed an electrophoretic homogeneity and was able to bind rotavirus particles in vitro. Rotavirus-PDI interaction was detected by capture ELISA using purified protein and rotavirus strains RRV and wild-type ECwt. Interaction between rotavirus particles and cellular PDI was detected by ELISA using cell lysates after virus inoculation. CONCLUSIONS: Rotavirus-PDI interaction was demonstrated in vitro as well as inMA104 cells and intestinal villi from suckling mice.

Impact of a probiotic fermented milk in the gut ecosystem and in the systemic immunity using a non-severe protein-energy-malnutrition model in mice.

BACKGROUND: Malnutrition affects the immune response, causing a decrease of defence mechanisms and making the host more susceptible to infections. Probiotics can reconstitute the intestinal mucosa and stimulate local and systemic immunity. The aim of this work was evaluate the effects of a probiotic fermented milk as a complement of a re-nutrition diet, on the recovery of the intestinal barrier, and mucosal and systemic immune functions in a murine model of non-severe protein-energy-malnutrition. Its potential protection against Salmonella enterica serovar Typhimurium (S. Typhimurium) infection was also analyzed. METHODS: Mice were undernourished and divided into 3 groups according to the dietary supplement received during re-nutrition (milk, probiotic fermented milk or its bacterial free supernatant) and compared to well-nourished and malnourished mice. They were sacrificed previous to the re-nutrition and 5 days post re-nutrition. The phagocytic activity of macrophages from spleen and peritoneum and the changes in the intestinal histology and microbiota were evaluated. Different immune cell populations and cytokine productions were analyzed in the small intestine tissues. The effect of the re-nutrition supplements on the systemic immunity using OVA antigen and against an infection with S. Typhimurium was also studied. RESULTS: Probiotic fermented milk was the most effective re-nutrition diet that improved the intestinal microbiota. Its administration also increased the number of IgA+ cells, macrophages and dendritic cells. The production of different cytokine (IFN-γ, TNF-α, IL-12) by these cells and the phagocytic activity in peritoneum and spleen was also increased. This re-nutrition diet also stimulated the systemic immune response against OVA antigen which was diminished after the malnutrition period and also improved the host response against S. Typhimurium, decreasing the spread of pathogenic bacteria to the liver and the spleen. The importance of the metabolites released during milk fermentation was also demonstrated through the analysis of the bacterial free supernatant obtained from the probiotic fermented milk, but the whole product showed the best effects in the parameters evaluated in this study. CONCLUSIONS: The administration of probiotic fermented milk as a dietary supplement during the re-nutrition process in a murine immunodeficiency model by malnutrition could be a good adjuvant diet to improve the gut and systemic immune response for the protection against Salmonella infection.

The glycan-binding protein galectin-1 controls survival of epithelial cells along the crypt-villus axis of small intestine.

Intestinal epithelial cells serve as mechanical barriers and active components of the mucosal immune system. These cells migrate from the crypt to the tip of the villus, where different stimuli can differentially affect their survival. Here we investigated, using in vitro and in vivo strategies, the role of galectin-1 (Gal-1), an evolutionarily conserved glycan-binding protein, in modulating the survival of human and mouse enterocytes. Both Gal-1 and its specific glyco-receptors were broadly expressed in small bowel enterocytes. Exogenous Gal-1 reduced the viability of enterocytes through apoptotic mechanisms involving activation of both caspase and mitochondrial pathways. Consistent with these findings, apoptotic cells were mainly detected at the tip of the villi, following administration of Gal-1. Moreover, Gal-1-deficient (Lgals1(-/-)) mice showed longer villi compared with their wild-type counterparts in vivo. In an experimental model of starvation, fasted wild-type mice displayed reduced villi and lower intestinal weight compared with Lgals1(-/-) mutant mice, an effect reflected by changes in the frequency of enterocyte apoptosis. Of note, human small bowel enterocytes were also prone to this pro-apoptotic effect. Thus, Gal-1 is broadly expressed in mucosal tissue and influences the viability of human and mouse enterocytes, an effect which might influence the migration of these cells from the crypt, the integrity of the villus and the epithelial barrier function.

Identificación de transportadores de glucosa intestinales mediante inmunohistoquímica y determinación de la absorción de glucosa durante el desarrollo y maduración del intestino delgado de crías de alpacas / Identification of intestinal glucose transporters by means of immunohistochemistry and determination of the glucose absorption during the development and maturation of the thin intestine of young of alpacas

Las alpacas, como otras especies animales, especialmente en la etapa de lactación, son susceptibles de sufrir afecciones que los conducen incluso a la muerte de no recibir la cantidad necesaria de energía en su dieta o adolecer de trastornos en el metabolismo de los carbohidratos. Es conocido que para que los monosacáridos pasen desde el lumen intestinal hacia la circulación sanguínea y luego a los tejidos es necesario que los enterocitos cuenten en su membrana con transportadores de glucosa como SGLT1 y GLUT2. En el presente estudio se ha evaluado la distribución de los transportadores de hexosas SGLT1 y GLUT2 en el eje cripta-vellosidad (zonas apical, media y cripta) de la mucosa intestinal del intestino delgado (duodeno, yeyuno e íleon) mediante la técnica de inmunohistoquímica (lHQ) así como los niveles de glucemia en crías de alpacas de 0-45 días de edad, las cuales fueron divididas en los siguientes grupos etáreos: 0 día, 1, 2, 3, 4, 5, 6 Y 7 semanas de edad. La evaluación correspondiente a la marcación por lHQ fue de 0 (sin marcación), 1 (marcación leve) y 2 (marcación fuerte); mientras que para la evaluación de la glucemia entre los grupos etáreos se usó la prueba de análisis de varianza de I sola vía con un nivel de confianza del 95 por ciento. Los resultados obtenidos muestran una marcación generalmente leve (1) desde el día 0 de edad para los 2 transportadores, la cual se hizo fuerte (2) hacia la semana 7, especialmente a nivel de yeyuno. Además, durante todo el estudio y en todas las porciones intestinales el SGLTI tuvo una marcación de mayor valor que la del GLUT2. Los niveles de glucemia mostraron valores entre 150±9.6 mg/dL (día de edad 0) y 176±12.5 mg/dL (semana 5 de edad), encontrándose diferencias estadísticas (p<0,05) entre los animales recién nacidos (crías sin ingesta de alimento) y las crías de las semanas 3, 4 Y 5 semana, que ya consumían leche y/o forraje. Se concluye que las proteínas transportadoras de glucosa SGLTI y GLUT2 están presentes desde el nacimiento lo que está relacionado a sus altos niveles de glucemia en este estadio y que la mayor presencia de SGLTl indicaría que su rol es el más importante en la etapa postnatal, siendo el yeyuno la porción de mayor absorción de monosacáridos

Alpacas, like other animal species, especially at the stage of lactation, are susceptible to conditions that could lead to death if they not receive the necessary amount of energy in their diet or suffer from disturbances in carbohydrate metabolismo It is known that the transport of monosaccharides from the intestinal lumen into the bloodstream and to the tissues requires the presence of the glucose transporters as SGLTl and GLUT2 in the membrane of the enterocytes. In the this study was evaluated the distribution of hexose transporters SGLTl and GLUT2 in the crypt-villus axis (apical, middle and crypt areas) of the intestinal mucosa of the small intestine (duodenum, jejunum and ileum) by immunohistochemistry (IHC), likewise the blood glucose levels in offspring of alpacas of 0-45 days old, which were divided the following age groups: 0 days, 1, 2, 3,4, 5,6 and 7 weeks. The assessment for dialing mc was 0 (no staining), 1 (mild staining) and 2 (strong staining). The comparative evaluation of blood glucose levels between the age groups was done using the analysis of variance test of 1 single track with a confidence level of 95 per cent. The results show mainly a mild staining (1) from day 0 of age for the 2 carriers, which becames strong (2) by week 7, especially at the jejunum level. In addition, throughout the study, the staining of SGLTl was greater than that of GLUT2. The blood glucose levels showed values between 150 ± 9.6 mg / dL (age day 0) and 176 ± 12.5 mg / dL (5 weeks old), which are statistical different (p<0.05) among newboms animals (babies with no food intake) and offspring of3, 4 and 5 weeks, which consumed food. Concluding that the glucose transporters SGLTl and GLUT2 are present since birth instant, which is related to the high blood glucose levels in this stage; the higher presence of the SGLTl seems to indicate that its function is the most important in the postnatal period, being the jejunum the portion of greatest absorption of monosaccharides