Ruminant livestock are important sources of human food and global greenhouse gas emissions. Feed degradation and methane formation by ruminants rely on metabolic interactions between rumen microbes and affect ruminant productivity. Rumen and camelid foregut microbial community composition was determined in 742 samples from 32 animal species and 35 countries, to estimate if this was influenced by diet, host species, or geography. Similar bacteria and archaea dominated in nearly all samples, while protozoal communities were more variable. The dominant bacteria are poorly characterised, but the methanogenic archaea are better known and highly conserved across the world. This universality and limited diversity could make it possible to mitigate methane emissions by developing strategies that target the few dominant methanogens. Differences in microbial community compositions were predominantly attributable to diet, with the host being less influential. There were few strong co-occurrence patterns between microbes, suggesting that major metabolic interactions are non-selective rather than specific.
Phytate breakdown in the digestive tract of broilers is affected by supplements of mineral phosphorus (P) and phytase with unknown interactions between the 2 factors. It was the objective to study phytate hydrolysis and the presence of inositol phosphate isomers (InsPs) as affected by supplements of mineral P and phytase in the small intestine of broilers. Fifteen-day old broilers were assigned to 48 pens of 20 broilers each (n = 8 pens/treatment). Two low-P corn-soybean meal-based diets without (BD-; 4.4 g P/kg dry matter) or with monocalcium phosphate (MCP; BD+; 5.2 g P/kg dry matter) were supplied without or with added phytase at 500 or 12,500 FTU/kg. On d 24, digesta from the duodenum/jejunum and lower ileum was pooled per segment on a by-pen basis, freeze-dried, and analyzed for P, InsPs, and the marker TiO2. Another 180 broilers (n = 6 pens/treatment, 10 birds each) were fed the 3 BD+ diets from d 1 to 21 to assess the influence of supplemented phytase on tibia mineralization and strength. Significant interactions between MCP and phytase supplements on myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) hydrolysis (duodenum/jejunum: P ≤ 0.001; ileum: P = 0.004) and level of specific lower InsPs were detected. Supplementation with 12,500 FTU/kg phytase resulted in 92% InsP6 hydrolysis and strong degradation of InsP5. This treatment resulted in higher P net absorption, affirmed by higher BW gain, tibia strength, and mineralization compared to treatments without or with 500 FTU/kg phytase (P ≤ 0.05). MCP supplementation reduced the degradation of InsP6 and specific lower InsPs in birds fed diets without or with 500 FTU/kg of phytase (P ≤ 0.05), but did not reduce InsP6 hydrolysis or degradation of InsP5 at the high phytase dose. Effects of added MCP on phytase efficacy depend on the dose of supplemented phytase. Differences in the concentrations of lower InsPs indicated that the initial step of InsP6 hydrolysis is not the only catabolic step that is influenced by MCP or phytase levels.
Molecular fingerprinting and sequencing based techniques have been widely used to characterize microbial communities. Terminal restriction fragment length polymorphism (T-RFLP) and 454-pyrosequencing were used to determine the microorganisms present in the different sections of the chicken gastrointestinal tract (GIT) (crop, jejunum, ileum and caeca). Broilers fed with diets differing in phosphorous (P) and calcium (Ca) as well as in phytase levels were used to study the microbiota of the upper and lower part of the GIT. A database with terminal restriction fragments (T-RF) of the most important organism present in the different gastrointestinal sections was constructed. The analysis revealed a distinct microbial assemblage on each section. Regardless of the diet, crop, jejunum and ileum were mainly colonized by Lactobacillaceae, and caeca were the most diverse site. The correlation between Lactobacillus crispatus and L. reuteri was positive in the crop, but negative in the jejunum. In crop samples, higher P and Ca levels led to a shift in the abundance of L. reuteri and L. crispatus to L. salivarius and L. taiwanensis whereas in the ileum supplementation of phytase favored L. salivarius and L. taiwanensis but resulted in decreased abundance of L. crispatus. Both methods were correlating significantly, being T-RFLP a reliable fingerprinting method to rapidly analyze large numbers of samples in a cost-effective and rapid manner. Results are easy to interpret with no need of deep bioinformatics knowledge and can be integrated with taxonomic information.
The objectives of this study were to determine the availability of P from mineral phosphate sources by using different basal diets and measurement of P retention and prececal (pc) P digestibility as well as pc myo-inositol phosphate (InsP) degradation in broilers. Semi-synthetic and corn-soybean meal-based basal diets were used in experiment 1, and corn-based and wheat-based basal diets were used in experiment 2. Anhydrous monosodium phosphate (MSPa) or monocalcium phosphate monohydrate (MCPh) was supplemented to increment the P concentration by 0.05, 0.10, and 0.15% or by 0.075 and 0.150% in experiments 1 and 2, respectively. Titanium dioxide was used as an indigestible marker. Diets were pelleted through a 3-mm screen. In experiment 1, retention was measured based on total excreta collection from 20 to 24 d of age using 7 replicated birds per diet. In experiment 2, digesta from the terminal ileum was collected from 22-d-old broilers penned in groups of 19 with 5 replicated pens per diet. The P retention response to supplemented MSPa did not differ between the 2 basal diets in experiment 1. The response in pc P digestibility to MCPh supplements also did not differ between the 2 basal diets in experiment 2, as calculated by linear regression analysis. Hydrolysis of InsP6 measured on both the excreta and pc levels was high in the basal diets without a mineral P supplement. Mineral P supplementation significantly decreased (P < 0.05) InsP6 hydrolysis from the InsP-containing diets in both experiments. Thus, the choice of the basal diet did not affect the evaluation of the supplemented mineral P source. However, calculated values for mineral P sources need to be adjusted for the decline in hydrolysis of InsP contained in the basal diet that results from the P supplement.
The responses of broilers to increments of dietary P concentration in P retention and P prececal digestibility were studied in 2 experiments using Ross 308 broilers in their fourth week of age. The low-P basal diet was mainly based on corn, potato protein, and corn starch. Titanium dioxide was used as the indigestible marker. Monobasic calcium phosphate was added in 6 (experiment 1) or 5 (experiment 2) graded levels up to a maximum of 8.1 g of P/kg of the diet. Five (experiment 1) and 6 (experiment 2) replicated pens of 10 birds were used. Excreta were collected from trays underneath the pens. Birds were asphyxiated by carbon dioxide exposure and the ileum (from Meckel's diverticulum to 2-cm anterior to the ileo-ceca-colonic junction) was dissected. Digesta was collected from the entire section (experiment 1) or from 3 subsections of equal lengths (experiment 2). Excretion of P increased linearly with increasing dietary P concentration up to a level of about 5.2 g of P/kg of the diet and increased nonlinearly with further increase in dietary P. In contrast, P flow in the ileum increased linearly over the entire range of P intake. Up to the level of 5.2 g of P/kg of the diet, P excretion was slightly lower than P flow in the ileum, but responses in P excretion and P flow in the ileum were similar, suggesting that P excretion with urine is very low and unaffected by P intake when the birds are supplied with P below their requirement. Between the 3 subsections of the ileum, calculated digestibility was significantly different for the basal diet and the diet with the 3 lowest levels of P supplementation. We concluded that the response in P prececal digestibility to increments in dietary P concentration is linear over a wider range of dietary P than the response in P retention. When digesta is collected from the ileum for determination of P prececal digestibility, the first third of the ileum should not be considered.
This study examined ruminal microbial community composition alterations during initial adaption to and following incubation in a rumen simulation system (Rusitec) using grass or corn silage as substrates. Samples were collected from fermenter liquids at 0, 2, 4, 12, 24, and 48 h and from feed residues at 0, 24, and 48 h after initiation of incubation (period 1) and on day 13 (period 2). Microbial DNA was extracted and real-time qPCR was used to quantify differences in the abundance of protozoa, methanogens, total bacteria, Fibrobacter succinogenes, Ruminococcus albus, Ruminobacter amylophilus, Prevotella bryantii, Selenomonas ruminantium, and Clostridium aminophilum. We found that forage source and sampling time significantly influenced the ruminal microbial community. The gene copy numbers of most microbial species (except C. aminophilum) decreased in period 1; however, adaption continued through period 2 for several species. The addition of fresh substrate in period 2 led to increasing copy numbers of all microbial species during the first 2–4 h in the fermenter liquid except protozoa, which showed a postprandial decrease. Corn silage enhanced the growth of R. amylophilus and F. succinogenes, and grass silage enhanced R. albus, P. bryantii, and C. aminophilum. No effect of forage source was detected on total bacteria, protozoa, S. ruminantium, or methanogens or on total gas production, although grass silage enhanced methane production. This study showed that the Rusitec provides a stable system after an adaption phase that should last longer than 48 h, and that the forage source influenced several microbial species.
The objectives of this study were to compare measurements of retention and prececal (pc) digestibility in evaluating mineral phosphorus (P) sources in 3- and 5-wk-old broilers. A corn-soybean meal-based basal diet was used (0.35% P on DM basis). Anhydrous monosodium phosphate (MSP(a)) or anhydrous dibasic calcium phosphate (DCP(a)) was supplemented to increment the P concentration by 0.08%, 0.16%, and 0.24%. Titanium dioxide was used as the indigestible marker. Two retention trials with excreta collection from d 16 to 20 and d 30 to 34 were conducted (n = 8 birds per diet). Another 8 pens of 10 birds from the same hatch were allocated to each diet on d 11 or 25 each to measure pc digestibility in both age periods. After 10 d of feeding, these birds were euthanized and the content of a defined section of the terminal ileum was obtained. Percentage P retention and pc digestibility for MSP(a) and DCP(a) were calculated by linear regression analysis. In 3-wk-old broilers, P retention for MSP(a) was 70% and significantly higher (P < 0.001) than for DCP(a) (29%). Values determined for pc digestibility at the same age were very similar (67% for MSP(a) and 30% for DCP(a); P < 0.001). In 5-wk-old broilers, P retention was 63% (MSP(a)) and 29% (DCP(a); P < 0.001) and pc digestibility was 54% (MSP(a)) and 25% (DCP(a); P = 0.002). We concluded that both retention and pc digestibility can be used for evaluating mineral P sources in broilers based on a regression approach. In 3-wk-old broilers, results obtained with both approaches were the same. In 5-wk-old broilers, the ranking of the 2 P sources was also the same for both approaches. Values did not differ significantly between the 2 age periods, but further studies on the relevance of broilers' age in P evaluation are suggested.
The chicken gastrointestinal tract (GIT) harbours a complex microbial community, involved in several physiological processes such as host immunomodulation and feed digestion. For the first time, the present study analysed dietary effects on the protein inventory of the microbiome in crop and ceca of broilers. We performed quantitative label-free metaproteomics by using 1-D-gel electrophoresis coupled with LC-MS/MS to identify the structural and functional changes triggered by diets supplied with varying amount of mineral phosphorous (P) and microbial phytase (MP). Phylogenetic assessment based on label-free quantification (LFQ) values of the proteins identified Lactobacillaceae as the major family in the crop section regardless of the diet, whereas proteins belonging to the family Veillonellaceae increased with the P supplementation. Within the ceca section, proteins of Bacteroidaceae were more abundant in the P-supplied diets, whereas proteins of Eubacteriaceae decreased with the P-addition. Proteins of the Ruminococcaceae increased with the amount of MP while proteins of Lactobacillaceae were more abundant in the MP-lacking diets. Classification of the identified proteins indicated a thriving microbial community in the case of P and MP supplementation, and stressed microbial community when no P and MP were supplied. Data are available via ProteomeXchange with identifier PXD003805.
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