Genomic characterization and gene regulation optimization to further improve an enzymatic mix used as feed additive

A common indigestible fraction of cereal grains, representing a large part of poultry diet, is their content in non-starch polysaccharides (NSP). Talaromyces versatilis is a filamentous fungus presenting the capability to secrete a mixture of enzymes used as animal feed additive (Rovabio® Excel) to enhance hydrolysis of plant cell wall polysaccharides. When incorporated to feed, the nutrients are more efficiently digested leading to a decreased need in agricultural products and hence a more sustainable production of poultry meat. In this context, the genome of T. versatilis was sequenced and annotated with a focus on genes likely to encode glycoside hydrolases, transcription factors and proteins involved in the secretion pathway. We also undertook a genome-wide transcriptome analysis, of the fungus exposed to glucose or milled wheat straw (a complex lignocellulosic material), using RNA-seq. The data revealed that, incubated on glucose and then transferred to wheat straw, the mycellium expressed differentially 926 genes between the two conditions. The differential response in gene expression of key mutants such as △xlnR, △creA or △araR were analysed in order to study their roles in regulating transcription. This approach provides a global view of the network that regulates the expression of the glycoside hydrolyse-encoding genes. More specifically, XlnR was identified as the transcription factor controling expression of genes involved in arabinoxylan degradation. Within the variety of NSP, arabinoxylan is the prominent type for wheat and corn (around 50%). Despite being mainly composed of xylose (X) and arabinose (A), the A:X ratio are different between corn and wheat, with a higher value for corn and a higher proportion of substituted xyloses compared to wheat. Arabinofuranosidase activity enhancement is key to attack arabinoxylans with a high A:X ratio which are recalcitrant to breakdown by single xylanase activity. Therefore, we aimed at improving the Rovabio® Excel in order to improve its capacity to degrade highly branched arabinoxylans, by enriching it in arabinofuranosidases and xylanases. To address such a goal while keeping its enzymatic diversity, we over-expressed the XlnR transcription factor. As a result, we obtained a modified strain of Talaromyces versatilis with on optimized genetic regulation to secrete a higher amount of arabinoxylan degrading enzymes. The resulting product, named Rovabio® Advance, tested in broilers allowed restoring nutrient availability, and so growth performance, even with a nutrient content diluted by 3% compared to a control diet

MHC-class-II are expressed in a subpopulation of human neural stem cells in vitro in an IFN gamma-independent fashion and during development

This work was supported by grants from Great Ormond Street Hospital Children’s Charity, Newlife Foundation, the Antony Nolan Trust, a studentship to CAG from Consejo Nacional de Ciencia y Tecnologia (CONACyT) and Instituto Jaliscience de la Juventud (IJJ), Mexico and GOSH NIHR Biomedical Research Centre. The human embryonic and fetal material was provided by the Human Developmental Biology Resource (http://hdbr.org) jointly funded by the Medical Research Council (grant G070089) and The Wellcome Trust (grant GR082557)

Results of an international phosphorus digestibility ring test with broiler chickens

The objective of this ring test was to investigate the prececal phosphorus (P) digestibility of soybean meal (SBM) in broiler chickens using the trial protocol proposed by the World's Poultry Science Association. It was hypothesized that prececal P digestibility of SBM determined in the collaborating stations is similar. Three diets with different inclusion levels of SBM were mixed in a feed mill specialized in experimental diets and transported to 17 collaborating stations. Broiler chicks were raised on commercial starter diets according to station-specific management routine. Then they were fed the experimental diets for a minimum of 5 d before content of the posterior half of the ileum was collected. A minimum of 6 experimental replicates per diet was used in each station. All diets and digesta samples were analyzed in the same laboratory. Diet, station, and their interaction significantly affected (P < 0.05) the prececal digestibility values of P and calcium of the diets. The prececal P digestibility of SBM was determined by linear regression and varied among stations from 19 to 51%, with significant differences among stations. In a subset of 4 stations, the prececal disappearance of myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate)-P; InsP6-P) also was studied. The prececal InsP6-P disappearance correlated well with the prececal P digestibility. We hypothesized that factors influencing InsP6 hydrolysis were main contributors to the variation in prececal P digestibility among stations. These factors were probably related to the feeding and housing conditions (floor pens or cages) of the birds in the pre-experimental phase. Therefore, we suggest that the World's Poultry Science Association protocol for the determination of digestible P be should extended to the standardization of the pre-experimental period. We also suggest that comparisons of P digestibility measurements among studies are made only with great caution until the protocol is more refined

Lipid bodies containing oxidatively truncated lipids block antigen cross-presentation by dendritic cells in cancer

Tumor-associated dendritic cells are defective in their ability to cross-present antigens, and they accumulate lipid bodies. Here the authors show that this defect is due to an impaired trafficking of peptide-MHC class I caused by the interaction of electrophilic lipids with chaperone heat shock protein 70

Effects of histocompatibility and host immune responses on the tumorigenicity of pluripotent stem cells

Pluripotent stem cells hold great promises for regenerative medicine. They might become useful as a universal source for a battery of new cell replacement therapies. Among the major concerns for the clinical application of stem cell-derived grafts are the risks of immune rejection and tumor formation. Pluripotency and tumorigenicity are closely linked features of pluripotent stem cells. However, the capacity to form teratomas or other tumors is not sufficiently described by inherited features of a stem cell line or a stem cell-derived graft. The tumorigenicity always depends on the inability of the recipient to reject the tumorigenic cells. This review summarizes recent data on the tumorigenicity of pluripotent stem cells in immunodeficient, syngeneic, allogeneic, and xenogeneic hosts. The effects of immunosuppressive treatment and cell differentiation are discussed. Different immune effector mechanisms appear to be involved in the rejection of undifferentiated and differentiated cell populations. Elements of the innate immune system, such as natural killer cells and the complement system, which are active also in syngeneic recipients, appear to preferentially reject undifferentiated cells. This effect could reduce the risk of tumor formation in immunocompetent recipients. Cell differentiation apparently increases susceptibility to rejection by the adaptive immune system in allogeneic hosts. The current data suggest that the immune system of the recipient has a major impact on the outcome of pluripotent stem cell transplantation, whether it is rejection, engraftment, or tumor development. This has to be considered when the results of experimental transplantation models are interpreted and even more when translation into clinics is planned

Digestible energy values of feed ingredients with or without addition of enzymes complex in growing pigs

The DE values and digestible nutrients content of 6 diets were measured in 60-kg male growing pigs fed restricted amount of feed. Diets were prepared from 5 ingredients [wheat (Triticum aestivum), corn (Zea mays), barley (Hordeum vulgare), wheat bran, and soybean (Glycine max) meal; inclusion levels of ingredients were not correlated] with or without carbohydrose enzyme (Rovabio Excel AP; 3300 endo-β-1,4-xylanase visco units and 300 endo-1,3(4)-β-glucanase units/kg of feed; 150 g/t of feed) according to a 6 × 2 factorial arrangement; dietary NDF ranged from 10.6 to 20.1% of DM. Pigs (5 per treatment) were placed in metabolism cages that allowed total collections of feces and urine for 10 d after a 11-d adaptation. Samples of feed, urine, and feces were analyzed for GE, ash, and N. Digestibility of GE, N, and OM were calculated. The effects of diet and enzyme (Enz) were evaluated by ANOVA. In addition, the DE and digestible nutrient contents of ingredients were calculated by regression of nutritive values of diets on level of ingredient inclusions. Apparent total tract digestibility of OM, N, and GE of diets were associated with dietary NDF content (r = –0.97; P < 0.001) and were increased (P < 0.05) by Enz addition by 0.4, 1.6, and 0.5%-units (a difference between two percentage values) for OM, N, and GE digestibility, respectively. Improvement in DE value due to Enz averaged 0.09 MJ/kg DM (15.11 vs. 15.02 MJ/kg DM; P < 0.05). The ADG (891 vs. 850 g/d; P < 0.05) was also increased by Enz addition. The calculated DE content without Enz addition averaged 16.3, 16.4, 14.9, 10.5, and 17.2 MJ/kg DM for wheat, corn, barley, wheat bran, and soybean meal, respectively. The Enz addition increased the DE value of ingredients similarly, but the best response was observed for wheat (0.33 MJ/kg DM)

Evaluation of phytase dose effect on performance, bone mineralization, and prececal phosphorus digestibility in broilers fed diets with varying metabolizable energy, digestible amino acids, and available phosphorus concentration

ABSTRACT: The nutritional composition of diets and the provision of exogenous phytases play important roles in animal performance. Therefore, we evaluated the individual and combined impact of metabolizable energy (ME), digestible lysine (dLys), available phosphorus (avP) and calcium (Ca), and phytase dose (1,000 or 2,000 FTU/kg) on the growth performance, feed efficiency, phosphorus digestibility, and bone ash content of broiler chickens from 10 to 42 d of age. Experimental diets were formulated in a Box-Behnken design to contain various levels of ME (11.9, 12.2, 12.54, or 13.1 MJ/kg), dLys (0.91, 0.93, 0.96, or 1.00%) and avP/Ca (0.12/0.47, 0.21/0.58, or 0.33/0.68%). The effect of phytase was expressed in terms of the extra nutrients released. The diets were formulated to have consistent phytate substrate contents (0.28% in average). Body weight gain (BWG) and feed conversion ratio (FCR) were described via polynomial equations (R2 = 0.88 and 0.52, respectively), with interconnections between variables (ME, dLys, and avP/Ca). No interaction was observed among variables (P > 0.05). Metabolizable energy was the most important factor affecting BWG and FCR (linearly; P < 0.001). Reducing ME content from 13.1 to 11.9 MJ/kg in control diet resulted in a 6.8% decrease in BWG and a 3.1% increase in FCR (P < 0.001). The dLys contents also affected performance linearly (P < 0.001), but to a lesser degree; BWG decreased by 160 g when the dLys was reduced by 0.09% units, while the same reduction in dLys increased the FCR by 0.108 points. The inclusion of phytase alleviated the negative effects on feed intake (FI), BWG, and FCR. Phytase improved phosphorus digestibility and bone ash content according to a quadratic relationship. When phytase was added, ME negatively affected FI (r = −0.82, P < 0.001), whereas the dLys content was correlated with FCR (r = −0.80, P < 0.001). Supplementing phytase allowed the reduction of ME, dLys, and avP–Ca in the diet without affecting performance. The addition of phytase increased of ME, dLys, and avP by 0.20 MJ/kg, 0.04 and 0.18% units for 1,000 FTU/kg and 0.4 MJ/kg, 0.06 and 0.20% units for 2,000 FTU/kg