The Duration of the Trial Influences the Effects of Mineral Deficiency and the Effective Phytase Dose in Broilers' Diets

[EN] Two trials varying in duration (short- and long-term) were conducted to evaluate the effects of providing deficient (NC) or sufficient (PC) Ca and P levels, and different doses of a new phytase (250, 500, and 1000 FTU/kg feed), in broiler feed on growth performance, nutrient digestibility and retention, and tibia mineralization. A total of 80 and 490 male chicks (Ross) of 21 and 1 days of age were used in the short- and long-term trials, respectively. In the long-term trial, chicks fed NC diets showed a lower (p < 0.05) average daily gain and feed intake compared to chicks fed PC and a greater (p < 0.05) feed conversion ratio compared to 500 and 1000 FTU/kg feed during the starting period. Regarding the effects on minerals¿ and nutrients¿ coefficients of retention, animals fed NC showed a significantly higher digestibility for P than those fed the PC diet in the long-term trial. Additionally, feeding 250 to 500 FTU/kg diets increased most of the nutrients¿ digestibility in the short-term but only P digestibility in the long-term trial. Tibia mineralization increased linearly with phytase addition (p < 0.05) only in the long-term trial. In conclusion, the effects of dietary mineral and phytase levels on growth performance are more noticeable in young animals. In addition, the duration of the trial is key due to a possible adaptation phenomenon of birds to low P supplementary levels.This research was funded by the Centre for the Development of Industrial Technology (CDTI), supported by the Ministry of Science and Innovation, Government of Spain.Javadi, M.; Cerisuelo, A.; Cambra López, M.; Macias-Vidal, J.; Donadeu, A.; Dupuy, J.; Carpintero, L.... (2022). The Duration of the Trial Influences the Effects of Mineral Deficiency and the Effective Phytase Dose in Broilers' Diets. Animals. 12(11):1-19. https://doi.org/10.3390/ani12111418119121

Estimation of phosphorous and nitrogen waste in rainbow trout (Oncorhynchus mykiss, Walbaum, 1792) diets including different inorganic phosphorous sources

[EN] Aquaculture effluents with high levels of phosphorus (P) and nitrogen (N) contribute to eutrophication in the aquatic ecosystem. The environmental impact of phosphorus and N aquaculture waste may be diminished by modifying diet ingredients that improve phosphorous (P) digestibility, and therefore, reduce the P in metabolic waste. The content of P in fishmeal is high (30 g/kg), but the inclusion of fishmeal in the diet is reducing due to its high costs and limited accessibility; therefore, the addition of an inorganic P source is necessary to ensure a satisfactory level of available P in fish diets. Consequently, the present study aimed to evaluate the effect of four different inorganic P sources on P digestibility and excretion in rainbow trout (Oncorhynchus mykiss), as one of the most relevant aquaculture species. Monosodium/monocalcium phosphate with 2% of sodium source presented a P digestibility similar to monoammonium phosphate, but with lower nitrogen and phosphorus excretion into the environment, which is advantageous from a nutritional, environmental and industrial point of view (biofilters and recirculation systems in fish farms). This study was conducted to evaluate the apparent availability and P and N excretion in rainbow trout (Oncorhynchus mykiss) using different inorganic phosphorus sources. With this goal, fish (153 +/- 14.1 g) fed four inorganic P sources were assayed: monoammonium phosphate (MAP, NH4H2PO4), monosodium/monocalcium phosphate (SCP-2%, AQphos+, NaH2PO4/Ca(H2PO4)(2)center dot H2O in proportion 12/88), monosodium/monocalcium phosphate (SCP-5%, NaH2PO4/Ca(H2PO4)(2)center dot H2O in proportion 30/70) and monocalcium phosphate (MCP, Ca(H2PO4)(2)center dot H2O). Phosphorus (P) digestibility, in diets that included MAP and SCP-2% as inorganic phosphorus sources, were significantly higher than for SCP-5% and MCP sources. In relation to the P excretion pattern, independent of the diet, a peak at 6 h after feeding was registered, but at different levels depending on inorganic P sources. Fish fed an MAP diet excreted a higher amount of dissolved P in comparison with the rest of the inorganic P sources, although the total P losses were lower in MAP and SCP-2% (33.02% and 28.13, respectively) than in SCP-5% and MCP sources (43.35% and 47.83, respectively). Nitrogen (N) excretion was also studied, and the fish fed an SCP-5% diet provided lower values (15.8%) than MAP (28.0%). When N total wastes were calculated, SCP-2% and SCP-5% showed the lowest values (31.54 and 28.25%, respectively). In conclusion, based on P and N digestibility and excretion, the SCP-2% diet showed the best results from a nutritional and environmental point of view.This study has been developed under the framework of the project entitled "Improvement of the nutritional quality of aquaculture feeds through the incorporation of microalgae hydrolysates enriched in probiotic microorganisms-ALQUABIOTIC," ITC-20181099, corresponding to the FEDER INNTERCONECTA 2018 call, funded by CDTI (Centre for the Development of Industrial Technology), supported by the Ministry of Science, Innovation and Universities and co-financed by FEDER funds (European Regional Development Fund) within the 2014-2020 Intelligent Growth Operational Program.Milián-Sorribes, MC.; Tomas-Vidal, A.; Peñaranda, D.; Carpintero, L.; Mesa, JS.; Dupuy, J.; Donadeu, A.... (2021). Estimation of phosphorous and nitrogen waste in rainbow trout (Oncorhynchus mykiss, Walbaum, 1792) diets including different inorganic phosphorous sources. Animals. 11(6):1-14. https://doi.org/10.3390/ani11061700S11411

Profiling of open chromatin in developing pig (Sus scrofa) muscle to identify regulatory regions

There is very little information about how the genome is regulated in domestic pigs (Sus scrofa). This lack of knowledge hinders efforts to define and predict the effects of genetic variants in pig breeding programs. To address this knowledge gap, we need to identify regulatory sequences in the pig genome starting with regions of open chromatin. We used the "Improved Protocol for the Assay for Transposase-Accessible Chromatin (Omni-ATAC-Seq)" to identify putative regulatory regions in flash-frozen semitendinosus muscle from 24 male piglets. We collected samples from the smallest-, average-, and largest-sized male piglets from each litter through five developmental time points. Of the 4661 ATAC-Seq peaks identified that represent regions of open chromatin, >50% were within 1 kb of known transcription start sites. Differential read count analysis revealed 377 ATAC-Seq defined genomic regions where chromatin accessibility differed significantly across developmental time points. We found regions of open chromatin associated with downregulation of genes involved in muscle development that were present in small-sized fetal piglets but absent in large-sized fetal piglets at day 90 of gestation. The dataset that we have generated provides a resource for studies of genome regulation in pigs and contributes valuable functional annotation information to filter genetic variants for use in genomic selection in pig breeding programs

Circulating microRNA Profiles during the Bovine Oestrous Cycle

Up to 50% of ovulations go undetected in modern dairy herds due to attenuated oestrus behavior and a lack of high-accuracy methods for detection of fertile oestrus. This significantly reduces overall herd productivity and constitutes a high economic burden to the dairy industry. MicroRNAs (miRNAs) are ubiquitous regulators of gene expression during both health and disease and they have been shown to regulate different reproductive processes. Extracellular miRNAs are stable and can provide useful biomarkers of tissue function; changes in circulating miRNA profiles have been reported during menstrual cycles. This study sought to establish the potential of circulating miRNAs as biomarkers of oestrus in cattle. We collected plasma samples from 8 Holstein-Friesian heifers on days Days 0, 8 and 16 of an oestrous cycle and analysed small RNA populations on each Day using two independent high-throughput approaches, namely, Illumina sequencing (n = 24 samples) and Qiagen PCR arrays (n = 9 sample pools, 3-4 samples / pool). Subsequently, we used RT-qPCR (n = 24 samples) to validate the results of high-throughput analyses, as well as to establish the expression profiles of additional miRNAs previously reported to be differentially expressed during reproductive cycles. Overall, we identified four miRNAs (let-7f, miR-125b, miR-145 and miR-99a-5p), the plasma levels of which distinctly increased (up to 2.2-fold, P < 0.05) during oestrus (Day 0) relative to other stages of the cycle (Days 8 and 16). Moreover, we identified several hundred different isomiRs and established their relative abundance in bovine plasma. In summary, our results reveal the dynamic nature of plasma miRNAs during the oestrous cycle and provide evidence of the feasibility of using circulating miRNAs as biomarkers of reproductive function in livestock in the future

Isolation and characterization of equine native MSC populations

Abstract Background In contrast to humans in which mesenchymal stem/stromal cell (MSC) therapies are still largely in the clinical trial phase, MSCs have been used therapeutically in horses for over 15 years, thus constituting a valuable preclinical model for humans. In human tissues, MSCs have been shown to originate from perivascular cells, namely pericytes and adventitial cells, which are identified by the presence of the cell surface markers CD146 and CD34, respectively. In contrast, the origin of MSCs in equine tissues has not been established, preventing the isolation and culture of defined cell populations in that species. Moreover, a comparison between perivascular CD146+ and CD34+ cell populations has not been performed in any species. Methods Immunohistochemistry was used to identify adventitial cells (CD34+) and pericytes (CD146+) and to determine their localization in relation to MSCs in equine tissues. Isolation of CD34+ (CD34+/CD146–/CD144–/CD45–) and CD146+ (CD146+/CD34–/CD144–/CD45–) cell fractions from equine adipose tissue was achieved by fluorescence-activated cell sorting. The isolated cell fractions were cultured and analyzed for the expression of MSC markers, using qPCR and flow cytometry, and for the ability to undergo trilineage differentiation. Angiogenic properties were analyzed in vivo using a chorioallantoic membrane (CAM) assay. Results Both CD34+ and CD146+ cells displayed typical MSC features, namely growth in uncoated tissue culture dishes, clonal growth when seeded at low density, expression of typical MSC markers, and multipotency shown by the capacity for trilineage differentiation. Of note, CD146+ cells were distinctly angiogenic compared with CD34+ and non-sorted cells (conventional MSCs), demonstrated by the induction of blood vessels in a CAM assay, expression of elevated levels of VEGFA and ANGPT1, and association with vascular networks in cocultures with endothelial cells, indicating that CD146+ cells maintain a pericyte phenotype in culture. Conclusion This study reports for the first time the successful isolation and culture of CD146+ and CD34+ cell populations from equine tissues. Characterization of these cells evidenced their distinct properties and MSC-like phenotype, and identified CD146+ cells as distinctly angiogenic, which may provide a novel source for enhanced regenerative therapies

Comprehensive analysis of blood cells and plasma identifies tissue-specific miRNAs as potential novel circulating biomarkers in cattle

Abstract Background The potential of circulating miRNAs as biomarkers of tissue function, both in health and disease, has been extensively demonstrated in humans. In addition, circulating miRNA biomarkers offer significant potential towards improving the productivity of livestock species, however, such potential has been hampered by the absence of information on the nature and source of circulating miRNA populations in these species. In addition, many miRNAs originally proposed as robust biomarkers of a particular tissue or disease in humans have been later shown not to be tissue specific and thus to actually have limited biomarker utility. In this study, we comprehensively analysed miRNA profiles in plasma and cell fractions of blood from cattle with the aim to identify tissue-derived miRNAs which may be useful as biomarkers of tissue function in this important food animal species. Results Using small RNA sequencing, we identified 92 miRNAs with significantly higher expression in plasma compared to paired blood cell samples (n = 4 cows). Differences in miRNA levels between plasma and cell fractions were validated for eight out of 10 miRNAs using RT-qPCR (n = 10 cows). Among miRNAs found to be enriched in plasma, we confirmed miR-122 (liver), miR-133a (muscle) and miR-215 (intestine) to be tissue-enriched, as reported for other species. Profiling of additional miRNAs across different tissues identified the human homologue, miR-802, as highly enriched specifically in liver. Conclusions These results provide novel information on the source of bovine circulating miRNAs and could significantly facilitate the identification of production-relevant tissue biomarkers in livestock. In particular, miR-802, a circulating miRNA not previously identified in cattle, can reportedly regulate insulin sensitivity and lipid metabolism, and thus could potentially provide a specific biomarker of liver function, a key parameter in the context of post-partum negative energy balance in dairy cows