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We previously reported a study on 288 broiler (Gallus gallus) chicks who received caffeine in water between days 3 and 42, at levels of 0, 6.25, 12.5, 25, 50 and 100 mg/kg body weight (BW)/day. In the previous report, we found that caffeine caused pulmonary hypertension (PH)-associated mortality in a significant minority (20%-30%) of birds, including right ventricular hypertrophy and ascites. We have also shown a significant upregulation of the serotonin transporter (SERT), troponin T2, adenosine A1 receptor (ADORA1) and phosphodiesterase 5A (PDE5) in chicken suffering from PH. Here, we examine the resistant (survived) chicks from the first study that had not died due to acute heart failure and did not have clinical signs of pulmonary hypertension. Our goal was to determine whether birds who lacked overt signs of disease had subclinical manifestations, including similar changes in gene expression, growth rates and altered systemic haemodynamics. We found that growth was significantly increased by caffeine consumption (p < 0.01) at low doses; however, dosage over 50 mg/BW/d had remarkable adverse effects on growth (p < 0.01). Blood pressure, troponin T2 and PDE5 gene expression were not significantly altered by caffeine administration (p > 0.05). However, SERT gene expression linearly increased with increasing caffeine dosage (p < 0.01). The impact of caffeine on ADORA1 gene expression was dose dependent and nonlinear. In conclusion, despite the significant effects of caffeine on birds' growth, no significant negative effects of caffeine were observed on the cardiovascular function of resistant chickens. This work provides valuable information for further study on different dosage of caffeine in an animal model.
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Pollos , Hipertensión Pulmonar , Animales , Cafeína/farmacología , Pollos/metabolismo , Expresión Génica , Hipertensión Pulmonar/veterinaria , Receptor de Adenosina A1/genética , Receptor de Adenosina A1/metabolismo , Troponina T/genéticaRESUMEN
Broiler chickens reared under heat stress (HS) conditions have decreased growth performance and show metabolic and immunologic alterations. This study aimed to evaluate the effect of supplementation with a standardized blend of plant-derived isoquinoline alkaloids (IQ) on the growth performance, protein catabolism, intestinal barrier function, and inflammatory status of HS-treated chickens. Three hundred sixty 0-day-old Ross 308 male broiler chickens were randomly distributed into 2 treatment groups: control diet (no additives) or diet supplemented with 100 ppm IQ. At day 14, the chicks in each diet group were further divided into 2 groups, each of which was reared under thermoneutral (TN) (22.4°C) or constant HS (33.0°C) conditions until day 42. Each group consisted of 6 replicates with 15 birds per replicate, and chickens were provided ad libitum access to water and feed. During days 15-21, the body weight gain (BWG) and feed intake (FI) were significantly lower in the HS treatment group than in the TN group, and feed conversion ratio was higher (P < 0.05); these factors were not alleviated by IQ supplementation. During days 22-42, the final BW, BWG, and FI of the HS birds were better among those administered IQ than those that were not (P < 0.05). HS treatment increased plasma lipid peroxide, corticosterone, and uric acid concentrations as well as serum fluorescein isothiocyanate-dextran, a marker of intestinal barrier function, and decreased plasma total protein content (P < 0.05). These changes were not observed in the IQ group, suggesting that IQ supplementation improved oxidative damage, protein catabolism, and intestinal barrier function of chickens under HS. Isoquinoline alkaloid supplementation inhibited the expression of intestinal inflammatory factors, IL-6, tumor necrosis factor-like factor 1A, and inducible nitric oxide synthase under HS treatment (P < 0.05). These results suggest that IQ supplementation can improve the growth performance of broiler chickens under HS conditions, which may be associated with amelioration of oxidative damage, protein catabolism, intestinal barrier function, and inflammation.
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Alcaloides/farmacología , Pollos/fisiología , Respuesta al Choque Térmico/fisiología , Intestinos/efectos de los fármacos , Isoquinolinas/administración & dosificación , Alcaloides/administración & dosificación , Alimentación Animal/análisis , Animales , Pollos/crecimiento & desarrollo , Dieta/veterinaria , Suplementos Dietéticos , Calor , Intestinos/fisiología , Isoquinolinas/química , MasculinoRESUMEN
The growth promoting effect of supplementing animal feed with antibiotics like tetracycline has traditionally been attributed to their antibiotic character. However, more evidence has been accumulated on their direct anti-inflammatory effect during the last two decades. Here we used a pig model to explore the systemic molecular effect of feed supplementation with sub therapeutic levels of oxytetracycline (OTC) by analysis of serum proteome changes. Results showed that OTC promoted growth, coinciding with a significant down regulation of different serum proteins related to inflammation, oxidation and lipid metabolism, confirming the anti-inflammatory mechanism of OTC. Interestingly, apart from the classic acute phase reactants also down regulation was seen of a hibernation associated plasma protein (HP-27), which is to our knowledge the first description in pigs. Although the exact function in non-hibernators is unclear, down regulation of HP-27 could be consistent with increased appetite, which is possibly linked to the anti-inflammatory action of OTC. Given that pigs are good models for human medicine due to their genetic and physiologic resemblance, the present results might also be used for rational intervention in human diseases in which inflammation plays an important role such as obesity, type 2 diabetes and cardiovascular diseases.
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Proteínas de Fase Aguda/análisis , Regulación hacia Abajo/efectos de los fármacos , Glicoproteínas/sangre , Oxitetraciclina/administración & dosificación , Porcinos/crecimiento & desarrollo , Alimentación Animal , Animales , Antibacterianos/sangre , Antiinflamatorios/administración & dosificación , Suplementos Dietéticos , Electroforesis , Haptoglobinas/análisis , Hibernación , Proteína Amiloide A Sérica/análisis , Espectrometría de Masas en TándemRESUMEN
BACKGROUND AND AIMS: Malnutrition and the use of Total Parenteral Nutrition (TPN) contribute considerably to hospital costs. Recently, we reported on the introduction of malnutrition screening and monitoring of TPN use in our hospital, which resulted in a large (40%) reduction in TPN and improved quality of nutritional care in two years (2011/12). Here, we aimed to assure continuation of improved care by developing a detailed malnutrition screening and TPN use protocol involving instruction tools for hospital staff, while monitoring the results in the following two years (2013/14). METHODS: A TPN decision tree for follow up of TPN in patients and a TP-EN instruction card for caregivers was introduced, showing TPN/EN introduction schedules based on the energy needs of patients according to EB guidelines, also addressing the risk of refeeding syndrome. TPN patients were monitored by dietitians and TPN usage and costs were presented to the (medical) staff. Screening and treatment of malnourished patients by dietitians is simultaneously ongoing. RESULTS: In 2014 48% of patients, hospitalized for at least 48 h, were screened on malnutrition, 17% of them were diagnosed at risk, 7.9% malnourished and treated by dietitians. TPN usage dropped by 53% and cost savings of 51% were obtained due to 50% decrease of TPN users in 2014 versus 2010. TPN over EN ratio dropped from 2.4 in 2010 to 1.2 in 2014. CONCLUSION: Sustained improvement of nutritional care and reduction of TPN usage and costs is possible by introduction of procedures embedded in the existing structures.
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Ahorro de Costo , Costos de Hospital , Apoyo Nutricional , Nutrición Parenteral Total/economía , Nutrición Parenteral Total/normas , Servicio de Admisión en Hospital/economía , Dietoterapia , Adhesión a Directriz , Hospitalización/economía , Hospitales , Humanos , Desnutrición/dietoterapia , Política Nutricional , Estado Nutricional , Grupo de Atención al PacienteRESUMEN
To study host-probiotic interactions in parts of the intestine only accessible in humans by surgery (jejunum, ileum and colon), pigs were used as model for humans. Groups of eight 6-week-old pigs were repeatedly orally administered with 5 × 10(12) CFU Lactobacillus plantarum 299v (L. plantarum 299v) or PBS, starting with a single dose followed by three consecutive daily dosings 10 days later. Gene expression was assessed with pooled RNA samples isolated from jejunum, ileum and colon scrapings of the eight pigs per group using Affymetrix porcine microarrays. Comparison of gene expression profiles recorded from L. plantarum 299v-treated pigs with PBS-treated pigs indicated that L. plantarum 299v affected metabolic and immunological processes, particularly in the ileum. A higher expression level of several B cell-specific transcription factors/regulators was observed, suggesting that an influx of B cells from the periphery to the ileum and/or the proliferation of progenitor B cells to IgA-committed plasma cells in the Peyer's patches of the ileum was stimulated. Genes coding for enzymes that metabolize leukotriene B4, 1,25-dihydroxyvitamin D3 and steroids were regulated in the ileum. Bioinformatics analysis predicted that these metabolites may play a role in the crosstalk between intestinal immune cells and sub-mucosal adipocytes. Together with regulation of genes that repress NFKB- and PPARG-mediated transcription, this crosstalk may contribute to tempering of inflammatory reactions. Furthermore, the enzyme adenosine deaminase, responsible for the breakdown of the anti-inflammatory mediator adenosine, was strongly down-regulated in response to L. plantarum 299v. This suggested that L. plantarum 299v-regulated production of adenosine by immune cells like regulatory T cells may also be a mechanism that tempers inflammation in the ileum, and perhaps also in other parts of the pig's body.
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The non-antibiotic anti-inflammatory theory of antimicrobial growth promoters (AGP) predicts that alternatives can be selected by simple in vitro tests. In vitro, the known AGP oxytetracycline (OTC) and a Macleaya cordata extract (MCE) had an anti-inflammatory effect with a half-maximal inhibitory concentration of 88 and 132 mg/l, respectively. In vivo, chickens received three different concentrations of MCE in drinking-water, OTC in feed and a control. Body weight (BW), feed intake (FI) and gain:feed (G:F) ratio were determined on days 14, 21 and 35. On day 35, body composition was determined. Plasma α1-acid glycoprotein (α1-AG) concentration was measured on days 21 and 35, and the expression of several jejunal inflammatory genes was determined on day 35. OTC-fed chickens showed a significantly higher BW, FI and G:F ratio compared with the control group at all time points. MCE had a significant linear effect on BW on days 21 and 35, and the G:F ratio was improved only over the whole period, whereas FI was not different. Only MCE but not OTC decreased the percentage of abdominal fat. Plasma α1-AG concentration increased from day 21 to 35, with the values being lower in the treatment groups. Both OTC and MCE significantly reduced the jejunal mucosal expression of inducible NO synthase. For most parameters measured, there was a clear linear dose-response to treatment with MCE. In conclusion, the results are consistent with the anti-inflammatory theory of growth promotion in production animals.
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Antiinflamatorios/farmacología , Pollos/crecimiento & desarrollo , Oxitetraciclina/farmacología , Papaveraceae/química , Extractos Vegetales/farmacología , Animales , Composición Corporal , Peso Corporal/efectos de los fármacos , Expresión Génica , Inflamación/genética , Interleucina-10/genética , Interleucina-1beta/genética , Mucosa Intestinal/metabolismo , Yeyuno/metabolismo , Masculino , Óxido Nítrico Sintasa de Tipo II/genética , Orosomucoide/análisis , Fitoterapia , Aumento de Peso/efectos de los fármacosRESUMEN
One of the major challenges in pig production is managing digestive health to maximize feed conversion and growth rates, but also to minimize treatment costs and to warrant public health. There is a great interest in the development of useful tools for intestinal health monitoring and the investigation of possible prophylactic/ therapeutic intervention pathways. A great variety of in vivo and in vitro intestinal models of study have been developed in the recent years. The understanding of such a complex system as the intestinal system (IS), and the study of its physiology and pathology is not an easy task. Analysis of such a complex system requires the use of systems biology techniques, like proteomics. However, for a correct interpretation of results and to maximize analysis performance, a careful selection of the IS model of study and proteomic platform is required. The study of the IS system is especially important in the pig, a species whose farming requires a very careful management of husbandry procedures regarding feeding and nutrition. The incorrect management of the pig digestive system leads directly to economic losses related suboptimal growth and feed utilization and/or the appearance of intestinal infections, in particular diarrhea. Furthermore, this species is the most suitable experimental model for human IS studies. Proteomics has risen as one of the most promising approaches to study the pig IS. In this review, we describe the most useful models of IS research in porcine and the different proteomic platforms available. An overview of the recent findings in pig IS proteomics is also provided.
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Mucosa Intestinal/metabolismo , Proteómica/métodos , Porcinos/genética , Animales , HumanosRESUMEN
Weaned piglets are very susceptible to diarrhea caused by enterotoxigenic Escherichia coli. In the past, various natural components were proposed to have beneficial effects by reducing the effects of diarrheal infectious diseases in humans and animals, and thus may represent an alternative for the use of (prophylactic) antibiotics. Alternatives may inactivate enterotoxigenic Escherichia coli heat-labile toxin (LT) by interfering with toxin binding to the cellular receptor GM1. In this study, various plants and other natural substances were tested for inhibitory properties, in the GM1 binding assay, and in the LT-induced cAMP production in Vero cells. The toxic dose of each compound was determined in a cell viability assay, and the highest nontoxic concentrations were used in the GM1 and cAMP assays. Results demonstrated that only d-(+)-galactose, lactose, N-acetyl-d-galactosamine, and two tea extracts were able to inhibit the binding of LT to its GM1 receptor. In the cAMP assay, only the two tea extracts showed inhibitory activity. This shows that d-(+)-galactose, lactose, and N-acetyl-d-galactosamine can indeed inhibit LT binding to GM1 based on structural homology with GM1 in the absence of living cells. However, in the cAMP assay, d-(+)-galactose, and lactose, N-acetyl-d-galactosamine are apparently metabolized to below their effective inhibitory concentration, likely predicting limited practical applicability in vivo. Both tea extracts maintained their activity in the presence of cells. The active compounds in both are probably polyphenols, which are not easily metabolized, and most likely work by aggregating the toxin. In conclusion, the combination of methods used here is a convenient and fast method for preselecting natural substances containing potentially toxin-binding compounds. Furthermore, if antidiarrhea activity is attributed to compounds found inactive here, their activity is unlikely based on interference with toxin binding.
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Toxinas Bacterianas/antagonistas & inhibidores , Diarrea/veterinaria , Escherichia coli Enterotoxigénica/efectos de los fármacos , Enterotoxinas/antagonistas & inhibidores , Infecciones por Escherichia coli/veterinaria , Proteínas de Escherichia coli/antagonistas & inhibidores , Receptores de Superficie Celular/metabolismo , Enfermedades de los Porcinos/prevención & control , Abrus/química , Acetilgalactosamina/farmacología , Animales , Toxinas Bacterianas/metabolismo , Canavalia/química , Supervivencia Celular , Chlorocebus aethiops , Diarrea/microbiología , Diarrea/prevención & control , Enterotoxinas/metabolismo , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/prevención & control , Proteínas de Escherichia coli/metabolismo , Fabaceae/química , Galactosa/farmacología , Humanos , Lactosa/farmacología , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Polifenoles/aislamiento & purificación , Polifenoles/farmacología , Unión Proteica , Porcinos , Enfermedades de los Porcinos/microbiología , Té/química , Factores de Tiempo , Células VeroRESUMEN
BACKGROUND: The aim of this study was to identify transcription factors/regulators that play a crucial role in steering the (innate) immune response shortly (within a few hours) after the first contact of the intestinal mucosa with an inflammatory mediator, and to test whether the processes regulated by these factors/regulators can be modulated by chemical substances of natural origin. METHODS: We experimentally induced inflammation by perfusion of surgically applied jejunal loops with Salmonella enterica subspecies enterica serovar Typhimurium DT104 in three pigs. Segments of mock and Salmonella treated loops were dissected after 2, 4 and 8 hours of perfusion. IL8 and IL1-beta mRNA expression levels were measured in mucosal scrapings of all segments. Furthermore, intra-animal microarray comparisons (isogenic) between Salmonella and mock treated segments after 8 hours, and inter-animal comparisons between similar Salmonella-treated loops of each pig at 2 and 4 hours, were performed. RESULTS: IL-1beta and IL8 mRNA levels, and intra-animal microarray comparisons at 8 hours between Salmonella and mock treated segments showed that the response-time and type of response to Salmonella was different in all three pigs. This plasticity allowed us to extract a comprehensive set of differentially expressed genes from inter-animal comparisons at 2 and 4 hours. Pathway analysis indicated that many of these genes play a role in induction and/or tempering the inflammatory response in the intestine. Among them a set of transcription factors/regulators known to be involved in regulation of inflammation, but also factors/regulators for which involvement was not expected. Nine out of twenty compounds of natural origin, which according to literature had the potential to modulate the activity of these factors/regulators, were able to stimulate or inhibit a Salmonella-induced mRNA response of inflammatory-reporter genes IL8 and/or nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha in cultured intestinal porcine epithelial cells. CONCLUSIONS: We describe a set of transcription factors/regulators possibly involved in regulation of "very early" immune mechanism which determines the inflammatory status of the intestine later on. In addition, we show that these mechanisms may be modulated by chemical substances of natural origin.
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Pollos , Fiebre/etiología , Mitocondrias/fisiología , Músculo Esquelético/fisiopatología , Enfermedades de las Aves de Corral/etiología , Animales , Bélgica/epidemiología , Femenino , Fiebre/mortalidad , Fiebre/fisiopatología , Masculino , Países Bajos/epidemiología , Enfermedades de las Aves de Corral/mortalidad , Enfermedades de las Aves de Corral/fisiopatologíaRESUMEN
Diarrhoea due to enterotoxigenic Escherichia coli with fimbriae F4 (ETEC-F4) is an important problem in neonatal and just weaned piglets and hence for the pig farming industry. There is substantial evidence for a genetic basis for susceptibility to ETEC-F4 since not all piglets suffer from diarrhoea after an ETEC-F4 infection. It is assumed that the wild boar was originally ETEC-F4 resistant and that susceptibility towards ETEC arose after domestication. There are different phenotypes in the pig determined by which of the three existing F4 variants (F4ab, F4ac or F4ad) they are susceptible or resistant for. This suggests that several F4 receptors exist, expressed individually or in combination with each other on the brush border of the piglet's small intestine. As such, the mucin-type glycoproteins (IMTGP) are described as F4ab/ac receptors, while the intestinal neutral glycospingolipid (IGLad) is proposed as an F4ad receptor. GP74 is a putative F4ab receptor. However, the specific genes that encode for the susceptibility are not yet known. In the past decades, linkage analyses revealed that the loci encoding for the receptor(s) for the two most frequent variants F4ab and F4ac were mapped to the 13th chromosome of the pig (Sus scrofa 13, SSC13). After fine mapping, the region of interest was mapped between two microsatellite markers, Sw207 and S0075, and interesting candidate genes surfaced. Numerous SNP analyses and a few expression studies on the three MUC-genes (MUC4, MUC13 and MUC20) and the transferrin receptor gene (TFRC) as well as on some other positional candidate genes have been performed in order to find the causative mutation for the ETEC-F4ab/ac receptor(s). However, until today, the exact mutation causing susceptibility to ETEC-F4 remains unknown.
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Escherichia coli Enterotoxigénica/fisiología , Infecciones por Escherichia coli/veterinaria , Fimbrias Bacterianas/fisiología , Enfermedades de los Porcinos/genética , Animales , Susceptibilidad a Enfermedades/microbiología , Susceptibilidad a Enfermedades/veterinaria , Infecciones por Escherichia coli/genética , Infecciones por Escherichia coli/microbiología , Mutación , Fenotipo , Porcinos , Enfermedades de los Porcinos/microbiologíaRESUMEN
Enterotoxigenic Escherichia coli (ETEC) strains that produce heat-stable (ST) and/or heat-labile (LT) enterotoxins are cause of post-weaning diarrhea in piglets. However, the relative importance of the different enterotoxins in host immune responses against ETEC infection has been poorly defined. In the present study, several isogenic mutant strains of an O149:F4ac(+), LT(+) STa(+) STb(+) ETEC strain were constructed that lack the expression of LT in combination with one or both types of ST enterotoxins (STa and/or STb). The small intestinal segment perfusion (SISP) technique and microarray analysis were used to study host early immune responses induced by these mutant strains 4 h after infection in comparison to the wild type strain and a PBS control. Simultaneously, net fluid absorption of pig small intestinal mucosa was measured 4 h after infection, allowing us to correlate enterotoxin secretion with gene regulation. Microarray analysis showed on the one hand a non-toxin related general antibacterial response comprising genes such as PAP, MMP1 and IL8. On the other hand, results suggest a dominant role for STb in small intestinal secretion early after post-weaning infection, as well as in the induced innate immune response through differential regulation of immune mediators like interleukin 1 and interleukin 17.
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Escherichia coli Enterotoxigénica/metabolismo , Enterotoxinas/química , Absorción , Animales , Cartilla de ADN/genética , Genotipo , Sistema Inmunológico , Interleucina-1/metabolismo , Interleucina-17/metabolismo , Interleucina-8/metabolismo , Mucosa Intestinal/metabolismo , Metaloproteinasa 1 de la Matriz/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Perfusión , Fenotipo , Programas Informáticos , Porcinos , Factores de TiempoRESUMEN
The high similarity between pigs and humans makes pigs a good gastrointestinal (GI) model for humans. Recently an epithelial cell line originating from the jejunum of pig (IPEC-J2) became available. Once validated, this model can be used to investigate the complex interactions occurring in the intestine. The advantages of using IPEC-J2 as in vitro model of the GI tract are the high resemblance between humans and pigs, and the ease of extrapolating in vitro to in vivo characteristics. In this study, the IPEC-J2 cells were functionally characterized by measuring the trans-epithelial electrical resistance (TEER), and by histological and ultrastructural studies. IPEC-J2 cells grown on six different permeable support systems, were investigated. The Transwell(®)-COL collagen-coated membrane (1.12 cm(2)) showed the best results concerning time efficiency and TEER values. The optimum seeding density of 12 × 10(5) cells/mL ensured that after 9 days of differentiation a confluent monolayer was formed. The decrease in TEER values after a maximum had been reached, coincided with the ultrastructural development of apical microvilli. We conclude that IPEC-J2 cells grown on collagen-coated membranes represent a valuable in vitro model system for the small intestinal epithelium which can be of great interest for intestinal research.
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Acute secretory diarrhea is a major cause of morbidity and mortality in young animals and humans. Deaths result from excessive fluid and electrolyte losses. The disease is caused by non-invasive bacteria such as Vibrio cholerae and Escherichia coli which produce enterotoxins, however, much less is known about the role of individual host responses. Here we report the response of intact porcine small intestinal mucosa to infection with enterotoxigenic E. coli (ETEC). Jejunal segments in four piglets were infused with or without ETEC, and perfused for 8h, and net absorption measured. Microarray analysis at 8h post-infection showed significant differential regulation of on average fifteen transcripts in mucosa, with considerable individual variation. Differential net absorption varied between animals, and correlated negatively with the number of up regulated genes, and with one individual gene (THO complex 4). This shows that quantitative differences in gene regulation can be functionally linked to the physiological response in these four animals.
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Escherichia coli Enterotoxigénica/fisiología , Infecciones por Escherichia coli/fisiopatología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Enfermedades Intestinales/microbiología , Enfermedades de los Porcinos/metabolismo , Animales , Antígenos de Neoplasias , Biomarcadores de Tumor , Northern Blotting , Interacciones Huésped-Patógeno , Intestino Delgado/metabolismo , Intestino Delgado/microbiología , Yeyuno/metabolismo , Yeyuno/microbiología , Lectinas Tipo C , Análisis por Micromatrices , Proteínas Asociadas a Pancreatitis , Porcinos , Enfermedades de los Porcinos/microbiologíaRESUMEN
IPEC-J2, a promising in vitro model system, is not well characterized especially on the transcriptional level, in contrast to human counterparts. The aim of this study was to characterize the gene expression in IPEC-J2 cells when coincubated with enterotoxigenic Escherichia coli (ETEC), nonpathogenic E. coli, and E. coli endotoxin. Apical infection of polarized IPEC-J2 monolayers caused a time-dependent decrease in transepithelial electrical resistance (TEER). Microarray analysis showed up-regulation of interleukins when IPEC-J2 were cocultured with E. coli strains this has so far never been measured in this cell line. Highest IL8 expression was found with the ETEC strain possessing the F4 fimbrium, suggesting IPEC-J2 cells to be F4 receptor positive, confirmed in a brush border membrane adhesion assay. It is concluded that the innate immune responses to pathogens and LPS makes the IPEC-J2 cell line a suitable model for research on intestinal host pathogen interaction.
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Host-microorganism interactions in the intestinal tract are complex, and little is known about specific nonpathogenic microbial factors triggering host responses in the gut. In this study, mannose-specific interactions of Lactobacillus plantarum 299v with jejunal epithelium were investigated using an in situ pig Small Intestinal Segment Perfusion model. The effects of L. plantarum 299v wild-type strain were compared with those of two corresponding mutant strains either lacking the gene encoding for the mannose-specific adhesin (msa) or sortase (srtA; responsible for anchoring of cell surface proteins like Msa to the cell wall). A slight enrichment of the wild-type strain associated with the intestinal surface could be observed after 8 h of perfusion when a mixture of wild-type and msa-mutant strain had been applied. In contrast to the mutant strains, the L. plantarum wild-type strain tended to induce a decrease in jejunal net fluid absorption compared with control conditions. Furthermore, after 8 h of perfusion expression of the host gene encoding pancreatitis-associated protein, a protein with proposed bactericidal properties, was found to be upregulated by the wild-type strain only. These observations suggest a role of Msa in the induction of host responses in the pig intestine.
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Mucosa Intestinal/microbiología , Yeyuno/microbiología , Lactobacillus plantarum/fisiología , Manosa/metabolismo , Adhesinas Bacterianas/genética , Adhesinas Bacterianas/fisiología , Aminoaciltransferasas/genética , Aminoaciltransferasas/fisiología , Animales , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/fisiología , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Cisteína Endopeptidasas/genética , Cisteína Endopeptidasas/fisiología , Absorción Intestinal , Yeyuno/fisiología , Lactobacillus plantarum/genética , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Masculino , Modelos Animales , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Proteínas Asociadas a Pancreatitis , Probióticos , PorcinosRESUMEN
Germ-free piglets were orally infected with virulent rotavirus to collect jejunal mucosal scrapings at 12 and 18 hours post infection (two piglets per time point). IFN-gamma mRNA expression was stimulated in the mucosa of all four infected piglets, indicating that they all responded to the rotavirus infection. RNA pools prepared from two infected piglets were used to compare whole mucosal gene expression at 12 and 18 hpi to expression in uninfected germ-free piglets (n=3) using a porcine intestinal cDNA microarray. Microarray analysis identified 13 down-regulated and 17 up-regulated genes. Northern blot analysis of a selected group of genes confirmed the data of the microarray. Genes were functionally clustered in interferon-regulated genes, proliferation/differentiation genes, apoptosis genes, cytoskeleton genes, signal transduction genes, and enterocyte digestive, absorptive, and transport genes. Down-regulation of the transport gene cluster reflected in part the loss of rotavirus-infected enterocytes from the villous tips. Data mining suggested that several genes were regulated in lower- or mid-villus immature enterocytes and goblet cells, probably to support repair of the damaged epithelial cell layer at the villous tips. Furthermore, up-regulation was observed for IFN-gamma induced guanylate binding protein 2, a protein that effectively inhibited VSV and EMCV replication in vitro (Arch Virol 150:1213-1220, 2005). This protein may play a role in the small intestine's innate defense against enteric viruses like rotavirus.
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Yeyuno/inmunología , Yeyuno/virología , Infecciones por Rotavirus/inmunología , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/virología , Animales , Proteínas de Unión al GTP/biosíntesis , Perfilación de la Expresión Génica , Vida Libre de Gérmenes , Interferón gamma/biosíntesis , Mucosa Intestinal/inmunología , Modelos Biológicos , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , PorcinosRESUMEN
Age-dependent changes in blood concentrations of four bovine acute phase proteins (APPs), serum amyloid A (SAA), lipopolysaccharide binding protein (LBP), haptoglobin (Hp) and alpha(1)-acid glycoprotein (AGP), were examined using two groups of newborn dairy calves. APP concentrations were monitored for either 3 weeks (Group A, n=13) or 2 months (Group B, n=13) after birth. Blood was collected at day 0-1 (Group A only), day 3 and then once or twice weekly until the end of the study. The possible transfer of colostral SAA as a source of SAA in the offspring was investigated by determining SAA isoforms in the serum of calves and in colostrum of their dams. Serum concentrations of all four APPs were high after birth, and concentrations showed a gradual decrease during the first 3 weeks of life. The lowest concentrations were at 21 days of age, after which concentrations stabilized. The calves synthesized adult SAA isotypes, and circulating SAA was not derived from colostrum. The results indicated that post-partum elevation of APPs is associated with the birth process and/or factors in colostrum and not necessarily with disease-related processes. This stresses the importance of considering a calf's age when interpreting APP concentrations in serum.
Asunto(s)
Proteínas de Fase Aguda/metabolismo , Bovinos/sangre , Factores de Edad , Animales , Animales Recién Nacidos , Western Blotting/veterinaria , Femenino , Focalización Isoeléctrica/veterinaria , Masculino , Isoformas de ProteínasRESUMEN
The objective of the present study was to evaluate the potential effects of dietary L-carnitine supplementation on acute phase protein response upon a lipopolysaccharide (LPS) challenge of male broiler chickens receiving a commercial broiler diet supplemented with 15 or 100 mg L-carnitine/kg or an unsupplemented (control) diet from 14 days of age onwards. At 28 days of age, eight chickens per dietary treatment were weighed and subcutaneously injected with 300 microg LPS from E. coli (100 microg LPS/ml saline) or 3 ml saline (unsupplemented group only). During the next 10 days, blood samples were taken repeatedly and analysed for their hemopexin (HX) and alpha-1 acid glycoprotein (AGP) levels. Extra dietary L-carnitine did not affect broiler performance. At day 1 postinjection, plasma HX and AGP levels were significantly increased in all treatment groups. However, the elevations in circulating HX and AGP levels were more pronounced in the L-carnitine supplemented chickens, especially in the 100mg L-carnitine group. It is concluded that extra L-carnitine in the diet of broiler chickens enhances or advances the acute phase protein response. The exact mode of action needs to be elucidated but seems to be consistent with a glucocorticoid mimicking effect.
Asunto(s)
Reacción de Fase Aguda/inducido químicamente , Carnitina/farmacología , Pollos/metabolismo , Dieta/veterinaria , Hemopexina/metabolismo , Lipopolisacáridos/farmacología , Orosomucoide/metabolismo , Alimentación Animal , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Carnitina/administración & dosificación , Pollos/crecimiento & desarrollo , Suplementos Dietéticos , Masculino , Factores de Tiempo , Aumento de Peso/efectos de los fármacosRESUMEN
Salmonella enterica serovar typhimurium (S. typhimurium) species are a leading cause of human invasive gastroenteritis. There is increasing in vitro evidence about Salmonella interaction with isolated cells or cell lines (macrophages, and enterocytes) on the molecular level, however, very little is known about in vivo interactions during actual invasion. We investigated the early interaction of S. typhimurium with intact small intestinal mucosa, in a pig model. Intestinal segments were infected with or without S. typhimurium DT104, and perfused. Whole mucosal gene expression was analyzed by cDNA array on 0, 2, 4, and 8h post-infection. Invasion resulted in the upregulation of only eight transcripts in jejunal mucosa, among those the proinflammatory IL-8 (at 4h only), and the antiinflammatory STAT3 (at 4 and 8h). The limited number of differentially expressed genes found here in vivo compared to in vitro is most likely due to the presence of multiple, heterogenous cell interactions in intact mucosa. Furthermore, it is concluded that S. typhimurium evades strong host responses by downregulating the local inflammatory response.