RESUMEN
Humans and their immune system are confronted with mold-contaminated food and/or mold-contaminated air in daily life and indoor activities. This results in metabolic stress and unspecific disease symptoms. Other studies provided evidence that exposure to mold is associated with the etiology of allergies. Deoxynivalenol (DON) is of great concern due to its frequent occurrence in toxically relevant concentrations. The exposure to this toxin is a permanent health risk for both humans and farm animals because DON cannot be significantly removed during standard milling and processing procedures. However, the direct effect on immunity or hematology is poorly defined because most investigations could not separate the effect of DON-contaminated feed intake. Due to the widespread distribution of DON after rapid absorption, it is not surprising that DON is known to affect the immune system. The immune system of the organism has one important function, to defend against the invasion of unknown substances/organisms. This study shows for the first time a synergistic effect of both-low physiological DON-doses in combination with low LPS-doses with the focus on the IL-8 expression on protein and RNA level. Both doses were found in vivo. IL-8 together with other anorectic cytokines like IL-1ß can affect the food intake and anorexia. We could also show that a calcium-response is not involved in the increased IL-8 production after acute DON stimulation with high or low concentrations.
Asunto(s)
Interleucina-8 , Monocitos , Transducción de Señal , Tricotecenos , Tricotecenos/toxicidad , Interleucina-8/metabolismo , Transducción de Señal/efectos de los fármacos , Monocitos/efectos de los fármacos , Monocitos/metabolismo , Animales , Biosíntesis de Proteínas/efectos de los fármacos , Humanos , Células CultivadasRESUMEN
A 10-week feeding experiment was carried out examining the effects of deoxynivalenol (DON)-contaminated maize treated with different sodium sulphite (SoS) concentrations on performance, health and DON-plasma concentrations in fattening pigs. Two maize batches were used: background-contaminated (CON, 0.73 mg/kg maize) and Fusarium-toxin contaminated (DON, 44.45 mg/kg maize) maize. Both were wet preserved at 20% moisture content, with one of three (0.0, 2.5, 5.0 g/kg maize) sodium sulphite concentrations and propionic acid (15%). Each maize batch was then mixed into a barley-wheat-based diet at a proportion of 10%, resulting in the following 6 feeding groups: CON- (CON + 0.0 g SoS/kg maize), CON2.5 (CON + 2.5 g SoS/kg maize), CON5.0 (CON + 5.0 g SoS/kg maize), DON- (DON + 0.0 g SoS/kg maize), DON2.5 (DON + 2.5 g SoS/kg maize) and DON5.0 (DON + 5.0 g SoS/kg maize). Dietary DON concentration was reduced by ~ 36% in group DON2.5 and ~ 63% in group DON5.0. There was no impact on ZEN concentration in the diets due to SoS treatment. Pigs receiving diet DON- showed markedly lower feed intake (FI) compared to those fed the control diets. With SoS-treatment of maize, FI of pigs fed the DON diet (DON5.0: 3.35 kg/d) were comparable to that control (CON-: 3.30 kg/day), and these effects were also reflected in live weight gain. There were some effects of SoS, DON or their interaction on serum urea, cholesterol and albumin, but always within the physiological range and thus likely negligible. SoS wet preservation of Fusarium-toxin contaminated maize successfully detoxified DON to its innocuous sulfonates, thus restoring impaired performance in fatteners.
Asunto(s)
Alimentación Animal/análisis , Contaminación de Alimentos/prevención & control , Sulfitos/farmacología , Tricotecenos/análisis , Aumento de Peso/efectos de los fármacos , Albúminas/análisis , Crianza de Animales Domésticos , Animales , Colesterol/sangre , Fusarium/patogenicidad , Porcinos , Urea/sangre , Zea mays/químicaRESUMEN
We studied the constancy of the relationship between rectal and intraabdominal temperature as well as their linkage to inflammatory markers (leucocyte counts, kynurenine-to-tryptophan ratio (Kyn-Trp ratio), tumour necrosis factor alpha (TNF-α) in healthy and in pigs exposed to lipopolysaccharide (LPS) and/or deoxynivalenol (DON). Barrows (n = 44) were fed 4 weeks either a DON-contaminated (4.59 mg DON/kg feed) or a control (CON) diet and equipped with an intraabdominal temperature logger and a multicatheter system (V.portae hepatis, V.lienalis, Vv.jugulares) facilitating infusion of 0.9% NaCl (CON) or LPS (7.5 µg/kg BW) and simultaneous blood sampling. Body temperatures were measured and blood samples taken every 15 min for leucocyte counts, TNF-α and Kyn-Trp ratio. Combination of diet and infusion created six groups: CON_CONjug .-CONpor. , CON_CONjug. -LPSpor. , CON_LPSjug. -CONpor. , DON_CONjug. -CONpor. , DON_CONjug. -LPSpor. , DON_LPSjug. -CONpor. . The relationship between both temperatures was not uniform for all conditions. Linear regression revealed that an intraabdominal increase per 1°C increase in rectal temperature was ~25% higher in all LPS-infused pigs compared to NaCl-infusion, albeit diet and site of LPS infusion modified the magnitude of this difference. Inflammatory markers were only strongly present under LPS influence and showed a significant relationship with body temperatures. For example, leucocyte counts in clinically inconspicuous animals were only significantly correlated to core temperature in DON-fed pigs, but in all LPS-infused groups, irrespective of diet and temperature method. In conclusion, the gradient between body core and rectal temperature is constant in clinically inconspicuous pigs, but not under various pathophysiological conditions. In the latter, measurement of inflammatory markers seems to be a useful completion.
Asunto(s)
Temperatura Corporal/efectos de los fármacos , Inflamación/veterinaria , Lipopolisacáridos/toxicidad , Tricotecenos/toxicidad , Alimentación Animal/análisis , Animales , Biomarcadores/sangre , Inflamación/inducido químicamente , Inflamación/metabolismo , Quinurenina/sangre , Porcinos , Tricotecenos/administración & dosificación , Triptófano/sangreRESUMEN
The objective of this experiment was to determine the effects of conjugated linoleic acid (CLA) and vitamin E as well as their interaction on performance variables and lipomobilization during late pregnancy and early lactation (wk 6 antepartum until wk 10 postpartum). For this purpose, 59 pluriparous German Holstein cows were assigned to 4 dietary groups in a 2 × 2 design with the factors CLA and vitamin E at 2 levels. For this trial, we selected cows with a high body condition score because they are more likely to mobilize fat and consequently are at a higher risk of developing ketosis. Furthermore, concentrate proportions were adjusted to provoke ketosis. Lactation performance variables were analyzed in 3 periods (d 42 antepartum until calving, 1 to 21 d in milk, 22 to 70 d in milk). Dry matter intake and net energy intake were reduced in animals receiving CLA. Milk fat content was reduced in the CLA group compared with the control group (4.83 vs. 5.46% in period 2; 3.36 vs. 4.57% in period 3). In the vitamin E and the CLA + vitamin E groups, reduction of milk fat content was observed in period 3 (3.76 vs. 4.57% compared with the control group). Milk yield was not affected by treatment. ß-Hydroxybutyrate concentrations and liver lipid contents were not influenced by CLA or vitamin E. Moreover, longitudinal changes of adipose tissue depot mass were not affected by dietary treatments. Results suggest that the effects CLA had on milk composition were compensated by an increased milk yield and a decreased dry matter intake. Reduced milk energy output in CLA-treated animals was compensated by a reduced dry matter intake. Therefore, the net energy balance was not affected by either treatment. Consequently, we found no group effect on the mobilization of adipose tissue.
Asunto(s)
Ácidos Linoleicos Conjugados/farmacología , Vitamina E/metabolismo , Animales , Bovinos , Dieta/veterinaria , Suplementos Dietéticos , Metabolismo Energético , Femenino , Lactancia/efectos de los fármacos , Leche/metabolismoRESUMEN
The porcine intestinal epithelium is a primary target for mycotoxin deoxynivalenol (DON) and lipopolysaccharides (LPS). Although epithelial cells are exposed to these toxins mainly from the luminal-chyme compartment an exposure from the blood side resulting from systemic absorption cannot be excluded. Thus, we investigated the effect of DON and LPS, alone or combined, on porcine intestinal epithelial cells IPEC-J2 on a transcriptional, translational and functional level when administered either from apical or basolateral. IPEC-J2 cells were cultured on 12-well inserts in complete medium at 5% CO2 and 39°C and subjected to following treatments: control (CON), 2000 ng/mL DON, 1 µg/mL LPS or DON+LPS for 72 h, either from apical or basolateral. Transepithelial electrical resistance (TEER), protein and IL-8 content were measured and microarray analysis, qRT-PCR (IL-8, zonula occludens-1 ZO-1, ß-actin), Western Blot (ZO-1, ß-actin) and immunofluorescence (ZO-1) were performed. Data of at least three independent experiments were analysed with ANOVA and Dunnett's post hoc test. Basolateral DON resulted in significantly lower cell counts (p<0.05) with larger cells (p<0.01), whereas apical DON reduced total (p<0.001) and specific protein content (IL-8 content CON vs. DON: 2378 pg/3 mL vs. 991 pg/3 mL; p<0.001). Transcripts of ß-actin and ZO-1 were significantly upregulated in response to DON, irrespective of direction, whereas IL-8 mRNA remained unaffected. However, ZO-1 spatial distribution in the tight junction and its function (TEER) were detrimentally affected by basolateral DON only. In conclusion, direction of DON exposure affected IPEC-J2 differently on a translational and functional level, but was mainly inconsequential on a transcriptional level.
Asunto(s)
Células Epiteliales/efectos de los fármacos , Lipopolisacáridos/toxicidad , Tricotecenos/toxicidad , Actinas/genética , Actinas/metabolismo , Animales , Recuento de Células , Células Cultivadas , Escherichia coli/química , Interleucina-8/genética , Interleucina-8/metabolismo , Intestinos/citología , Intestinos/efectos de los fármacos , Porcinos , Uniones Estrechas/efectos de los fármacos , Uniones Estrechas/metabolismo , Regulación hacia Arriba , Proteína de la Zonula Occludens-1/genética , Proteína de la Zonula Occludens-1/metabolismoRESUMEN
The numerous pores in the basement membrane (BM) of the intestinal villi are essential for the communication of enterocytes with cells in the lamina propria, an important mechanism for the induction of intestinal immune responses. The intestinal epithelial barrier is affected by the mycotoxin deoxynivalenol (DON) from both the apical (luminal) and basolateral (serosal) side. The pig is the most susceptible species to the anorectic and immune-modulating effects of DON, which is most prevalent in crops. We analysed in pigs the effect of DON-contaminated feed on the composition and perforation of the BM and the presence of CD16(+) cells or their dendrites in the epithelium. In addition to in vivo experiments, in vitro studies were carried out. Using microarray analyses, the effects of DON on IPEC-J2 cells were studied with the focus on the BM. Our in vivo results showed in the control pigs: (1) a significant increased pore number (p ≤ 0.001) in the jejunum in comparison to ileum, (2) no difference in the pore size, and (3) comparable frequency of intraepithelial CD16(+) cells/dendrites in the jejunum and ileum. There was a marked trend that DON feeding increases: (1) the pore number in jejunum, and (2) the number of CD16(+) cells/dendrites in the epithelium (Tukey-Kramer; p = 0.055 and p = 0.067, respectively). The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells). The down-regulation of genes like syndecan, fibulin 6 and BM-40 was observed. These proteins are important factors in the BM composition and in formation of pores. Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells. Thus, DON affects the composition of the BM.
Asunto(s)
Membrana Basal/inmunología , Mucosa Intestinal/inmunología , Proteínas de la Membrana/inmunología , Porcinos/inmunología , Tricotecenos/farmacología , Alimentación Animal/toxicidad , Animales , Membrana Basal/citología , Membrana Basal/ultraestructura , Western Blotting , Línea Celular , Movimiento Celular/inmunología , Células Epiteliales , Contaminación de Alimentos , Mucosa Intestinal/citología , Mucosa Intestinal/ultraestructura , Análisis de los Mínimos Cuadrados , Masculino , Proteínas de la Membrana/genética , Microscopía Confocal/veterinaria , Microscopía Fluorescente/veterinaria , Análisis de Secuencia por Matrices de Oligonucleótidos/veterinaria , ARN/química , ARN/genéticaRESUMEN
The in vivo effects of the Fusarium toxin deoxynivalenol (DON) on albumin and fibrinogen synthesis in pigs and metabolic activity of porcine peripheral blood mononuclear cells (PBMCs) were studied alone or in combination with lipopolysaccharides (LPSs) in order to examine proposed synergistic effects of both substances. A total of 36 male castrated pigs (initial weight of 26 kg) were used. Uncontaminated (Control) and naturally DON-contaminated (chronic oral DON, 3.1mg/kg diet) wheat was fed for 37 days. On the day of protein synthesis measurement, pigs recruited from the Control group were treated once intravenously with (iv) DON (100 µg/kg live weight (LW)/h), iv LPS (7.5 µg/kgLW/h) or a combination of both substances, and six pigs from the chronic oral group were treated once with iv LPS. A treatment with DON alone exhibited no alterations of acute phase protein synthesis and metabolic activity of PBMC. There was no evidence that the chosen dosing regimen of DON had influences on the induced sub-acute stage of sepsis, as the LPS challenge, irrespective of DON co-exposure, mediated an acute phase reaction with a typical decrease of albumin synthesis, as well as changes in cytokine concentration and a loss of metabolic activity in PBMC.
Asunto(s)
Proteínas de Fase Aguda/biosíntesis , Citocinas/metabolismo , Leucocitos Mononucleares/efectos de los fármacos , Lipopolisacáridos/farmacología , Micotoxinas/farmacología , Porcinos/metabolismo , Tricotecenos/farmacología , Albúminas/metabolismo , Alimentación Animal , Animales , Fibrinógeno/metabolismo , Contaminación de Alimentos , Fusarium/química , Leucocitos Mononucleares/metabolismo , Masculino , Tricotecenos/análisis , Triticum/química , Triticum/microbiologíaRESUMEN
We investigated a proposed synergistic effect of deoxynivalenol (DON) and lipopolysaccharides (LPS) on small intestinal architecture and epithelial barrier integrity in pigs. Crypt depth and intestinal cell proliferation were analyzed, as well as expression of zonula occludens protein-1 (ZO-1) and ß-catenin of the apical junction complex along the small intestine. Barrows (26.2±4.1 kg) were fed restrictedly either a control diet (CON) or a diet naturally contaminated with 3.1 mg DON/kg feed (DON) for 37 d. At d 37, the control group was infused for 1 h either with 100 µg/kg BW of DON (CON-DON, n=6), 7.5 µg/kg BW of LPS (CON-LPS, n=6), a combination of both (CON-DON+LPS, n=7), or 0.9% NaCl (CON-CON, n=6) and the DON group with 7.5 µg/kg BW of LPS (DON-LPS, n=8) or 0.9% NaCl (DON-CON, n=6). Pigs were euthanized 3.25 h after start of infusion. Immunohistochemistry (5'-bromo-2'-deoxyuridine for proliferation) and immunofluorescence (ZO-1 and ß-catenin) from duodenum, proximal jejunum, mid-jejunum, proximal ileum, and terminal ileum were analyzed for crypt depth, cell proliferation, and apical junction proteins. Duodenal crypts were deeper compared with the other 4 intestinal regions, and proximal jejunal crypts were deeper than those of mid-jejunum and proximal ileum (P<0.001). Epithelial proliferation showed a bell-shaped distribution along the small intestinal axis. Duodenal proliferating cells had the least number compared with jejunal sections and proximal ileum (P<0.001). Neither DON nor LPS affected these variables. Zonula occludens-1 displayed a distinct spatial distribution in the epithelium with an apical and a cytosolic component. Apical expression of ZO-1 was severely damaged in the mid-jejunum (P<0.001) of CON-DON compared with animals treated with LPS. Also, in all animals receiving LPS systemically, the cytosolic ZO-1 fraction in the 3 upper gut sections disappeared completely. This effect was independent of DON presence. Control pigs had a greater basolateral ß-catenin accumulation (P<0.05) in the cells, whereas the protein distribution did not differ in CON-DON pigs. In conclusion, results of this experiment demonstrated that epithelial proliferation has a distinct pattern along the small intestine and is not necessarily positively linked to crypt depth in pigs. Furthermore, results indicate that LPS changed the spatial distribution of ZO-1. A synergistic effect of DON and LPS on intestinal architecture could not be verified in the present study.
Asunto(s)
Células Epiteliales/efectos de los fármacos , Escherichia coli/metabolismo , Mucosa Intestinal/citología , Lipopolisacáridos/farmacocinética , Porcinos/fisiología , Tricotecenos/farmacocinética , Animales , Proliferación Celular , Interacciones Farmacológicas , Células Epiteliales/citología , Regulación de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/toxicidad , Masculino , Tricotecenos/toxicidad , Proteína de la Zonula Occludens-1/genética , Proteína de la Zonula Occludens-1/metabolismo , beta Catenina/genética , beta Catenina/metabolismoRESUMEN
A total of 200 (Large White × Landrace) sows were used in a 39-day study to evaluate the effects of feeding a non-starch polysaccharide (NSP)-hydrolysing enzyme multicomplex (Rovabio(®) Excel) in conjunction with a high- or reduced nutrient-density diet during lactation on sow body condition, feed intake and progeny performance. Eight sows were selected each week for 25 weeks, blocked by parity and BW into groups of four, and within the block randomly assigned to one of the four treatments (n = 50/treatment). Treatments were: (1) LND: low energy (13.14 MJ of DE/kg), low CP (15%) diet; (2) LND + RE: LND with 50 mg/kg NSP-hydrolysing enzyme; (3) HND: high energy (14.5 MJ of DE/kg), high CP (16.5%) diet; and (4) HND + RE: HND with 50 mg/kg NSP-hydrolysing enzyme. Sows were fed treatment diets from day 109 of gestation until the day of subsequent service. Between weaning and re-service, Rovabio(®) Excel addition to LND diets resulted in an increase in energy intake; however, a reduction was observed when supplemented to the HND diet (P < 0.05). The inclusion of Rovabio(®) Excel increased feed and energy intake during week 3 (days 15 to 21) of lactation (P < 0.05). Sows fed diets supplemented with Rovabio(®) Excel had greater back-fat depth at weaning and service (P < 0.05); however, the magnitude of change in back-fat depth during lactation and from farrowing to service was not different between treatments. Feeding the HND diet increased energy intake before farrowing, throughout lactation and during the weaning to service interval (P < 0.01); however, overall, average daily feed intake tended to be reduced (P < 0.10). At service, sows fed the HND diet were heavier than sows fed the LND diet (P < 0.05); however, the magnitude of change in BW between treatments was not different. Feeding the HND diet to sows resulted in a tendency for heavier piglets at birth (P = 0.10) that tended to grow at a faster rate and be heavier at weaning than piglets from sows fed the LND diet (P = 0.06). These results indicate that NSP-degrading enzymes offer minimal benefit to sows and their progeny when fed before and during lactation; however, increasing energy intake of sows during lactation may beneficially affect progeny.
Asunto(s)
Alimentación Animal/análisis , Composición Corporal , Suplementos Dietéticos/análisis , Ingestión de Energía , Complejos Multienzimáticos/metabolismo , Porcinos/fisiología , Crianza de Animales Domésticos , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Dieta/veterinaria , Conducta Alimentaria , Femenino , Lactancia , Complejos Multienzimáticos/administración & dosificación , Polisacáridos/metabolismo , Porcinos/crecimiento & desarrolloRESUMEN
The specific function of the epithelium as critical barrier between the intestinal lumen and the organism's internal microenvironment is reflected by permanent maintenance of intercellular junctions and cellular polarity. The intestinal epithelial cells are responsible for absorption of nutritional components, facing mechanical stress and a changing oxygen supplementation via blood stream. Oxygen itself can regulate the barrier and the absorptive function of the epithelium. Therefore, we compared the dish cell culture, the transwell-like membrane culture and the oxygen enriched air-liquid interface (ALI) culture. We demonstrated strong influence of the different culture conditions on morphology and function of intestinal porcine epithelial cell lines in vitro. ALI culture resulted in a significant increase in cell number, epithelial cell layer thickness and expression as well as apical localisation of the microvilli-associated protein villin. Remarkable similarities regarding the morphological parameters were observed between ALI cultures and intestinal epithelial cells in vivo. Furthermore, the functional analysis of protein uptake and degradation by the epithelial cells demonstrated the necessity of sufficient oxygen supply as achieved in ALI cultures. Our study is the first report providing marked evidence that optimised oxygen supply using ALI cultures directly affects the morphological differentiation and functional properties of intestinal epithelial cells in vitro.
Asunto(s)
Diferenciación Celular , Células Epiteliales/citología , Mucosa Intestinal/citología , Citoesqueleto de Actina/metabolismo , Aire , Animales , Células Cultivadas , Células Epiteliales/metabolismo , Microscopía Electrónica de Transmisión de Rastreo , Microvellosidades/metabolismo , Oxígeno , PorcinosRESUMEN
Piglets, separated from their dam at 12 days of age and fed a milk substitute hourly, were used as a model for suckling. Animals were fitted with a terminal ileal T-cannula and a jugular vein catheter. At 28 days of age, half of the pigs had a dietary change to a cereal-based weaner diet fed as slurry, and the others remained on milk substitute. Animals were labelled by oral administration of 15N-labelled yeast for 10 days (days 15 to 25). Blood samples were taken twice a day to monitor 15N enrichment of the blood plasma. Diets included polyethylenglycol (PEG 4000) to allow calculation of apparent ileal digestibility of nitrogen and individual amino acids. Ileal bacterial nitrogen was calculated from D-alanine content of the digesta. Furthermore, small intestinal (SI) villus height and crypt depth were measured. Feed intake was increased by the dietary change. The total nitrogen flow was 3.2 ± 0.4 g/day and 5.9 ± 0.4 for the milk and weaner diet, respectively. Endogenous nitrogen flow at the terminal ileum was similar for both groups (milk diet 2.4 ± 0.4 v. weaner diet 2.2 ± 0.3 g/day), whereas the bacterial nitrogen content (0.08 ± 0.01 g/day milk diet v. 0.15 ± 0.01 g/day weaner diet, P < 0.01) and exogenous nitrogen flow (0.94 ± 0.16 g/day milk diet v. 3.29 ± 0.12 g/day weaner diet, P < 0.001) increased significantly in the weaner-diet group. The ileal apparent digestibility coefficient of protein was 0.81 ± 0.06 and 0.68 ± 0.01 for the milk replacer and the weaner diet, respectively. Morphology measurements made along the SI at 25%, 50% and 75% were similar between piglets fed milk replacer and those fed a cereal-based weaner diet. The only statistical effect (P < 0.01) of dietary change was an increase in crypt depth in the weaner-diet group. In conclusion, pigs, following a dietary change analogous to weaning, lack the capacity to fully digest a standard weaner diet. This may result in an increased nutrient content entering the large intestine and an altered microbiota. In the absence of a period of anorexia, often associated with traditional weaning, we saw no evidence of villous atrophy, but report here a significant crypt hyperplasia, especially at the 75% level, as a result of dietary change.