Intestinal epithelium integrity after delayed onset of nutrition in broiler chickens
Hollemans, M.S. ; Baal, J. van; Vries Reilingh, G. de; Kemp, B. ; Lammers, A. ; Vries, S. de - \ 2020
Poultry Science (2020). - ISSN 0032-5791 - 10 p.
broiler - delayed nutrition - gene expression - intestinal integrity - tight junction
Fasting older broiler chickens (>7 d of age) enlarges the intestinal tight junction (TJ) pore size, resulting in high paracellular intestinal permeability. Broiler chickens often do not receive feed and water (nutrition) directly after hatch, which may result in fasting up to 72 h of age. Whether perinatal fasting affects intestinal permeability is minimally studied. We therefore investigated whether delayed access to nutrition after hatch increases intestinal permeability, compared with broilers receiving early access to nutrition. Therefore, 432 hatched broilers received nutrition 72 h after hatch (delayed nutrition [DN]) or directly after hatch (early nutrition [EN]) and were reared under similar conditions until 14 d of age. Two hours after application of an oral pulse dose (3.85 mg) of fluorescein isothiocyanate-dextran (4000 Da) at 4, 10, and 14 d of age, blood plasma concentrations of the marker were measured in 24 to 36 broilers per treatment and time point. Marker concentration in plasma did not differ between DN and EN broilers at any age. The villus width measured in at least 8 broilers per treatment was smaller in DN than in EN broilers at 4 d for both the ileum (92 ± 3 μm vs. 121 ± 4; P < 0.001) and colon (100 ± 3 vs. 120 ± 4; P < 0.01). Real-time quantitative PCR revealed that the expression of TJ protein claudin 3 in the ceca was elevated in DN, compared with EN broilers at 4 d of age, whereas that of zonula occludens 1 in the ileum was reduced. Expression of host defense-related genes was reduced in DN, compared with EN broilers, in the ileum (cyclo-oxygenase 2, mucin 2) and ceca (interleukin 1β, cyclo-oxygenase 2). We conclude that 72-hour DN reduced the BW up to 14 d of age, coinciding with transient effects on the villus width in the ileum and colon, and divergent expression of genes involved in TJ formation and host defense. These effects likely reflect the delayed onset of intestinal and immune development in DN, compared with EN broilers, while DN does not fundamentally alter intestinal permeability.
Training of primary chicken monocytes results in enhanced pro-inflammatory responses
Verwoolde, Michel B. ; Biggelaar, Robin H.G.A. van den; Baal, Jürgen van; Jansen, Christine A. ; Lammers, Aart - \ 2020
Veterinary Sciences 7 (2020)3. - ISSN 2306-7381
Flow cytometry - Inflammatory response - Innate immune memory - Macrophages - Primary chicken monocytes - Β-glucan
Beta-glucan-stimulated mammalian myeloid cells, such as macrophages, show an increased responsiveness to secondary stimulation in a nonspecific manner. This phenomenon is known as trained innate immunity and is important to prevent reinfections. Trained innate immunity seems to be an evolutionary conserved phenomenon among plants, invertebrates and mammalian species. Our study aimed to explore the training of primary chicken monocytes. We hypothesized that primary chicken monocytes, similar to their mammalian counterparts, can be trained with β-glucan resulting in increased responses of these cells to a secondary stimulus. Primary blood monocytes of white leghorn chickens were primary stimulated with β-glucan microparticulates (M-βG), lipopolysaccharide (LPS), recombinant chicken interleukin-4 (IL-4) or combinations of these components for 48 h. On day 6, the primary stimulated cells were secondary stimulated with LPS. Nitric oxide (NO) production levels were measured as an indicator of pro-inflammatory activity. In addition, the cells were analyzed by flow cytometry to characterize the population of trained cells and to investigate the expression of surface markers associated with activation. After the secondary LPS stimulation, surface expression of colony stimulating factor 1 receptor (CSF1R) and the activation markers CD40 and major histocompatibility complex class II (MHC-II) was higher on macrophages that were trained with a combination of M-βG and IL-4 compared to unstimulated cells. This increased expression was paralleled by enhanced NO production. In conclusion, this study showed that trained innate immunity can be induced in primary chicken monocytes with β-glucan, which is in line with previous experiments in mammalian species. Innate immune training may have the potential to improve health and vaccination strategies within the poultry sector.
Innate immune training and metabolic reprogramming in primary monocytes of broiler and laying hens
Verwoolde, Michel B. ; Biggelaar, Robin H.G.A. van den; Vries Reilingh, Ger de; Arts, Joop A.J. ; Baal, Jürgen van; Lammers, Aart ; Jansen, Christine A. - \ 2020
Developmental and Comparative Immunology 114 (2020). - ISSN 0145-305X
Chickens - Immune training - Lipopolysaccharide - Metabolism - Monocytes - β-glucan
Recently, we have reported trained innate immunity in laying chicken monocytes. In the present study, we further investigated trained innate immunity of monocytes in layers and broilers. Monocytes of both breeds isolated from blood were trained in vitro with β-glucan, rec-chicken IL-4 or a combination of both, and restimulated with lipopolysaccharide (LPS), after which inflammation and metabolism-related responses were measured. Training of laying and broiler hen monocytes resulted in increased mRNA levels of IL-1β, iNOS and HIF-1α, but enhanced surface expression of CD40 and NO production was only observed in layers. Our in vitro study demonstrates that monocytes from different genetic backgrounds can be trained. However, the observed differences suggest a differential effect on immune functionality associated with innate training. Whether these differences in immune functions between layers and broilers have effect on disease resistance remains to be elucidated.
Coarse limestone does not alleviate the negative effect of a low Ca/P ratio diet on characteristics of tibia strength and growth performance in broilers
Hu, Y.X. ; Bikker, P. ; Duijster, M. ; Hendriks, W.H. ; Baal, J. van; Krimpen, M.M. van - \ 2020
Poultry Science 99 (2020)10. - ISSN 0032-5791 - p. 4978 - 4989.
broilers - Ca/P ratio - particle size of limestone - phosphorus and calcium digestibility
The hypothesis was tested that an increased digestion of coarse compared with fine limestone can alleviate the negative effects of a low dietary Ca/P ratio on the growth performance and characteristics of tibia strength (CTS) in broilers. A total of 1,152 Ross 308 broiler chickens received a standard commercial starter feed from day 0 to 13. From day 14 onward, birds received 1 of 12 diets containing 1 of 6 Ca/P ratios (0.50, 0.75, 1.00, 1.25, 1.50, and 1.75) and 1 of 2 limestone particle sizes (<500 [fine] and 500 to 2,000 [coarse] μm) in a study with a 6 × 2 factorial arrangement of treatments. Total P content was fixed at 5.5 g/kg for all treatment diets. Each treatment was replicated 6 times with 16 birds per replicate pen. On day 20 and 21, twelve birds per pen were randomly selected from 4 of the 6 replicate pens for tibia analysis and digesta collection from different gut segments. The apparent Ca digestibility was higher for fine than coarse limestone in the jejunum (P = 0.043). However, this difference in Ca digestibility disappeared for the low, whereas it remained for the high Ca/P ratios in the proximal (Pinteraction = 0.067) and distal (Pinteraction = 0.052) ileum. In addition, coarse limestone improved apparent P digestibility in the proximal and distal ileum (P < 0.001) but not in the jejunum (P = 0.305). Regardless of limestone particle size, reducing dietary Ca/P ratio linearly improved apparent Ca and P digestibility in the proximal and distal ileum (P < 0.001). Moreover, decreasing dietary Ca/P ratio linearly (P < 0.001) and quadratically (P < 0.046) reduced the CTS. Reducing dietary Ca/P ratio linearly (P < 0.003) and quadratically (P ≤ 0.006) decreased body weight gain and increased feed conversion ratio. For both fine and coarse limestone, the optimal Ca/P ratio was 1.00 to 1.25 to optimize apparent Ca and P digestibility while maintaining growth performance and CTS. Reducing Ca/P ratio from 1.75 to 1.00 improved distal ileal Ca and P apparent digestibility from 36.6 to 53.7% and 48.0 to 58.3%, respectively. In conclusion, coarse limestone is equally digestible with fine limestone at a low Ca/P ratio but is less digestible at a high Ca/P ratio, and the optimal Ca/P ratio in the diet is 1.00 to 1.25 for both fine and coarse limestone.
Impact of post-ruminally infused macronutrients on bovine mammary gland expression of genes involved in fatty acid synthesis, energy metabolism, and protein synthesis measured in RNA isolated from milk fat
Nichols, Kelly ; Bannink, André ; Baal, Jurgen Van; Dijkstra, Jan - \ 2020
Journal of Animal Science and Biotechnology 11 (2020)1. - ISSN 1674-9782
Cytoplasmic crescent - Endoplasmic reticulum biogenesis - Mammary cell - Milk fat globule - Milk synthesis - Tricarboxylic acid cycle
Background: Characterising the regulation of milk component synthesis in response to macronutrient supply is critical for understanding the implications of nutritional interventions on milk production. Gene expression in mammary gland secretory cells was measured using RNA isolated from milk fat globules from 6 Holstein-Friesian cows receiving 5-d abomasal infusions of saline, essential amino acids (AA), or glucose (GG) or palm olein (LG) without (LAA) or with (HAA) essential AA, according to a 6 × 6 Latin square design. RNA was isolated from milk fat samples collected on d 5 of infusion and subjected to real-time quantitative PCR. We hypothesised that mRNA expression of genes involved in de novo milk fatty acid (FA) synthesis would be differently affected by GG and LG, and that expression of genes regulating transfer of tricarboxylic acid cycle intermediates would increase at the HAA level. We also hypothesised that the HAA level would affect genes regulating endoplasmic reticulum (ER) homeostasis but would not affect genes related to the mechanistic target of rapamycin complex 1 (mTORC1) or the integrated stress response (ISR) network. Results: Infusion of GG did not affect de novo milk FA yield but decreased expression of FA synthase (FASN). Infusion of LG decreased de novo FA yield and tended to decrease expression of acetyl-CoA carboxylase 1 (ACC1). The HAA level increased both de novo FA yield and expression of ACC1, and tended to decrease expression of mitochondrial phosphoenolpyruvate carboxykinase (PCK2). mRNA expression of mTORC1 signaling participants was not affected by GG, LG, or AA level. Expression of the ϵ subunit of the ISR constituent eukaryotic translation initiation factor 2B (EIF2B5) tended to increase at the HAA level, but only in the presence of LG. X-box binding protein 1 (XBP1) mRNA was activated in response to LG and the HAA level. Conclusions: Results show that expression of genes involved in de novo FA synthesis responded to glucogenic, lipogenic, and aminogenic substrates, whereas genes regulating intermediate flux through the tricarboxylic acid cycle were not majorly affected. Results also suggest that after 5 d of AA supplementation, milk protein synthesis is supported by enhanced ER biogenesis instead of signaling through the mTORC1 or ISR networks.
Effect of dietary Ca content and microbial phytase inclusion on Ca and P excretion and growth performance in growing pigs
Hu, Y.X. ; Krimpen, M.M. van; Resink, J.W. ; Hendriks, W.H. ; Baal, J. van; Bikker, P. - \ 2020
In: Wias Annual Conference 2020 WIAS - p. 85 - 85.
Sixty growing pigs (Hypor Libra*Maxter, body weight 30.4±1.3 kg) were used to test the hy-pothesis that the negative effect of dietary high Ca content on P absorption was greater in microbial phytase supplemented diets than in phytase-free diets. Pigs were arranged in a 2×3 factorial block design including 2 levels of phytase (0 vs 500 FTU/kg microbial phytase) and 3 levels of dietary Ca content (1.8, 5.3 and 8.8 g/kg). The total P content was fixed at 4.7 g/kg for all diets. Treatments were applied to pigs for 21 days including a 4-d balance trial period (d 14-17) to quantitatively collect the feces and urine. Results indicated that the incremental dietary Ca content enhanced while phytase inclusion reduced fecal Ca and P excretion (P<0.001). Compared to the low Ca content, high Ca content increased fecal P excretion by 0.3 g/kg in phytase-free diets and by 0.7 g/kg in microbial phytase-supplemented diets (Pinteraction<0.001). The negative effect of the high dietary Ca content on P ab-sorption in microbial phytase supplemented diets, therefore, was about 2-2.5 times as great as in in phytase-free diets. Phytase enhanced the P absorption by 1.5, 1.2 and 1.1 g/kg in the low, medium and high Ca diets, respectively. Hence the phytase efficacy to improve P absorption was about 50% greater in the low Ca diet compared to the high Ca diets. As for growth performance in the entire experimental period (d0-20), the incremental Ca lev-el impaired (P=0.016) while phytase improved (P=0.025) the average daily gain. Microbial phytase inclusion also reduced the feed:gain ratio at high Ca diets (2.01 vs 1.80) but not at low Ca diets (1.77 vs 1.74, Pinteraction=0.067). In conclusion, the negative effect of the dietary high Ca content on total tract P absorption was greater in microbial phytase-supplemented diets than in phytase-free diets; phytase efficacy to improve P absorption was greater in low Ca diets than in high Ca diets.
|The influence of genetic background on trained innate immunity in chicken macrophages
Verwoolde, M.B. ; Biggelaar, Robin H.G.A. van den; Baal, J. van; Jansen, Christine A. ; Lammers, A. - \ 2019
|Effect of limestone particle size and Ca/P ratio on tibia quality and growth performance in broilers
Hu, Yixin ; Bikker, P. ; Duijster, M. ; Hendriks, W.H. ; Baal, J. van; Krimpen, M.M. van - \ 2019
In: Proceedings of the 44th Animal Nutrition Research Forum. - - p. 14 - 15.
Short communication: Relationship between lysine/methionine ratios and glucose levels and their effects on casein synthesis via activation of the mechanistic target of rapamycin signaling pathway in bovine mammary epithelial cells
Wang, F. ; Baal, J. van; Ma, L. ; Loor, J.J. ; Wu, Z.L. ; Dijkstra, J. ; Bu, D.P. - \ 2019
Journal of Dairy Science 102 (2019)9. - ISSN 0022-0302 - p. 8127 - 8133.
amino acid - casein - glucose - mammary cell signaling
The synthesis of protein requires the availability of specific AA and a large supply of energy in bovine mammary epithelial cells (BMEC). Whether an interaction exists between Lys/Met ratio and glucose level on milk protein synthesis and its potential regulatory mechanism is unclear. We investigated the effects of different Lys/Met ratios and glucose levels on casein synthesis-related gene expression in BMEC to elucidate the underlying molecular mechanisms. Primary BMEC were subjected to 4 treatments for 36 h, arranged in a 2 × 2 factorial design with Lys/Met ratios of 3:1 (1.2:0.4 mM, LM3.0; total AA = 8.24 mM) and 2.3:1 (1.4:0.6 mM, LM2.3; total AA = 8.64 mM) and glucose levels of 17.5 mM (high glucose level) and 2.5 mM (low glucose level). No interactions between Lys/Met ratio and glucose level on cell viability, cell cycle progression, mRNA, or protein expression levels were found. High glucose level increased cell proliferation and promoted cell cycle transition from intermediate phase (G1 phase) to synthesis (S phase) by approximately 50%, whereas Lys/Met ratio had no effect. Both mRNA and protein abundance of αS1-casein and β-casein were positively affected by LM3.0, whereas a high glucose level increased protein abundance of αS1-casein and β-casein and increased gene expression of CSN1S1 but not of CSN2. Furthermore, high glucose increased the mRNA abundance of ELF5 and decreased that of GLUT8, enhanced protein expression of total and phosphorylated mechanistic target of rapamycin (mTOR), and decreased phosphorylated AMP-activated protein kinase (AMPK) levels. Treatment LM3.0 had a stimulatory effect on total and phosphorylated mTOR but did not affect AMPK phosphorylation. The mRNA levels of JAK2, ELF5, and RPS6KB1 were upregulated and mRNA levels of EIF4EBP1 were downregulated with LM3.0 compared with LM2.3. Our results indicate that casein synthesis was regulated by Lys/Met ratio via JAK2/ELF5, mTOR, and its downstream RPS6KB1 and EIF4EBP1 signaling. In contrast, glucose regulated casein synthesis through promoting cell proliferation, accelerating cell cycle progression, and activating the ELF5 and AMPK/mTOR signaling pathways. Within the range of substrate levels in the present study, a change in Lys/Met ratio had a stronger effect on abundance of αS1-casein and β-casein than a change in glucose level.
|Trained innate immunity in chicken macrophages
Verwoolde, M.B. ; Biggelaar, Robin H.G.A. van den; Baal, J. van; Jansen, Christine A. ; Lammers, A. - \ 2019
In: Trade-offs in science – keeping the Balance. - Wageningen University & Research - p. 27 - 27.
|In vitro model to study trained innate immunity in chicken primary monocytes
Verwoolde, M.B. ; Biggelaar, Robin H.G.A. van den; Baal, J. van; Jansen, Christine A. ; Lammers, A. - \ 2018
|Low phosphorous diets in gestation and lactation stimulate calcium absorption in sows and reduce calcium and phosphorous absorption in offspring
Bikker, P. ; Liesegang, A. ; Baal, J. van - \ 2018
- p. 66 - 66.
|Both dietary copper(I)oxide and copper sulphate stimulate growth performance in pigs but differentially affect copper absorption and metal transporter genes
Bikker, P. ; Durosoy, S. ; Romeo, A. ; Baal, J. van - \ 2018
- p. 65 - 65.
|Differential effects of dietary copper sulphate and copper(I)oxide on gut microbiota of weaned piglets
Sinche Ambrosio, Carmen ; Smidt, H. ; Baal, J. van; Romeo, A. ; Bikker, P. - \ 2018
|Effect of dietary Ca to P ratio and particle size of limestone on growth performance and digesta pH in different intestinal segments in broilers
Hu, Yixin ; Bikker, P. ; Duijster, M. ; Baal, J. van; Hendriks, W.H. ; Krimpen, M.M. van - \ 2018
- p. 45 - 46.
|Glucose and ratio of lysine to methionine stimulate expression levels of genes involved in casein synthesis in bovine mammary epithelial cells
Wang, Fang ; Niu, J.L. ; Loor, J. ; Dijkstra, J. ; Baal, J. van; Bu, D.P. - \ 2017
- p. 82 - 83.
Effect of supplemental concentrate during the dry period or early lactation on rumen epithelium gene and protein expression in dairy cattle during the transition period
Dieho, K. ; Baal, J. van; Kruijt, L. ; Bannink, A. ; Schonewille, J.T. ; Carreño, D. ; Hendriks, W.H. ; Dijkstra, J. - \ 2017
Journal of Dairy Science 100 (2017)9. - ISSN 0022-0302 - p. 7227 - 7245.
Gene and protein expression - Rumen adaptation - Rumen epithelium - VFA absorption and metabolism
We previously reported 2 experiments with rumen-cannulated Holstein-Friesian dairy cows showing that during the transition period, rumen papillae surface area, and fractional absorption rate of volatile fatty acids (VFA) increase after calving. However, supplemental concentrate during the dry period and rate of increase of concentrate allowance during lactation affected papillae surface area, but not VFA absorption. Here we report the changes in gene and protein expression in rumen papillae related to tissue growth and VFA utilization. The lactation experiment treatment consisted of a rapid [RAP; 1.0 kg of dry matter (DM)/d; n = 6] or gradual (GRAD; 0.25 kg of DM/d; n = 6) increase of concentrate allowance (up to 10.9 kg of DM/d), starting at 4 d postpartum (pp). The dry period experiment treatment consisted of 3.0 kg of DM/d of concentrate (n = 4) or no concentrate (n = 5) during the last 28 d of the dry period. Real-time quantitative PCR analysis of rumen papillae showed that the expression of apoptosis-related genes was neither affected by day nor its interaction with treatment for both experiments. Expression of epithelial transporter genes was not affected by day or treatment in the lactation experiment, except for NBC1. In the dry period experiment, expression of MCT1, NBC1, DRA, NHE2, NHE3, and UT-B generally decreased after calving. A day and treatment interaction was observed for ATP1A1 in the dry period experiment, with greater expression at 18 and 8 d antepartum for concentrate than no concentrate. Generally, expression of VFA metabolism-related genes was not affected by day or its interaction with treatment. In the lactation experiment, immunoblotting of 5 selected genes showed that protein expression of DRA and PCCA was greater at 16 d pp compared with 3 and 44 d pp. Expression of NHE2 was greater, and that of ATP1A1 lower, at 16 and 44 d pp compared with 3 d pp, suggesting alterations in intracellular pH regulation and sodium homeostasis. Both MCT1 and PCCA protein were upregulated by RAP from 3 to 16 d pp, indicating modulations in VFA metabolism. Our data suggests that VFA absorption and metabolic capacity changed little per unit of surface area during the transition period, and suggests that a change in mitosis rate rather than apoptosis rate is associated with the increased ruminal VFA production, resulting in tissue growth. A significant but weak correlation between the examined gene and protein expression levels was observed only for PCCA, indicating that care must be taken when interpreting results obtained at either level.
Dose-dependent effects of copper supplementation of nursery diets on growth performance and fecal consistency in weaned pigs
Bikker, P. ; Jongbloed, A.W. ; Baal, J. van - \ 2016
Journal of Animal Science 94 (2016)7 supplement 3. - ISSN 0021-8812 - p. 181 - 186.
Copper - Fecal consistency - Growth performance - Pigs
Pharmacological levels of dietary copper (150–250 mg/kg) improve growth performance in pigs, but the majority of Cu is excreted in the manure and may contribute to accumulation in the soil. This experiment with 4 levels of dietary Cu supplementation (15, 80, 120, and 160 mg/kg) from weaning to d 56 after weaning was conducted to determine the dose-dependent effects of dietary Cu on growth performance and fecal consistency in pigs. From d 56 to slaughter, all pigs received the same grower and finisher diets supplemented with 15 mg/kg Cu. Each treatment comprised 10 pens with 8 pigs each, mixed males and females, as experimental units. The pigs were housed in a nursery unit until d 40 after weaning and in a grow–finish unit thereafter. Growth performance and fecal consistency were determined in 2-wk intervals until d 56; thereafter, growth performance was monitored between d 56 and slaughter. The Cu supplementation linearly increased ADFI and ADG (P <0.001) until d 40 and linearly increased G:F (P <0.01) until d 28 of the nursery period. Copper supplementation linearly increased ADG (P = 0.024) from d 40 to 56. The proportion of piglets with loose feces and diarrhea was reduced with incremental dietary Cu in each 2-wk interval until d 56. The combination of our data with 5 published studies in weaned pigs and subsequent regression analysis showed that the inclusion of the European Union maximum Cu supplement of 160 mg/kg enhances the ADG by approximately 14%. Reduction of the Cu supplement to 120 mg/kg would reduce the ADG by 2 to 3% and increase the number of piglets with loose feces.
|Influence of dietary copper content on the expression of copper absorption-related genes in the digestive tract of weaned pigs
Bikker, P. ; Jongbloed, A. ; Baal, J. van - \ 2015
In: Book of Abstracts 13th Digestive Physiology in Pigs Symposium. - - p. 209 - 209.
Copper in diets for weaned pigs : influence of level and duration of copper supplementation
Bikker, P. ; Baal, J. van; Binnendijk, G.P. ; Diepen, J.T.M. van; Troquet, L.M.P. ; Jongbloed, A.W. - \ 2015
Wageningen : Wageningen UR Livestock Research (Livestock Research report 830) - 38
varkens - biggen - spenen - koper - diervoedering - minerale supplementen - groei - dunne darm - voedertoevoegingen - biggenvoeding - pigs - piglets - weaning - copper - animal feeding - mineral supplements - growth - small intestine - feed additives - piglet feeding
This study was conducted to determine the influence of level (15-160 mg/kg voer)and period (2-8 weeks) of supply of a Cu-supplement on growth performance and expression of Cu absorption-related genes in different segments of the small intestine of weaned pigs.