Interaction between the host and the enteric microbiome is highly complex. Microbial involvement in certain pathologies is moderately well established, but the contribution of the microbiome to ...animal welfare, behavior, sustainability, immune development, nutritional status, physiology, and maturation is less clear. A valuable experimental model to enable scientists to explore the role of the microbiome in various domains is to compare various phenotypes of a conventionally reared (CV) cohort with those in a germ-free (GF) state. A GF animal is one that is devoid of any detectable microbial life including bacteria, viruses, protozoa and parasites. The GF state is different from gnotobiotic animals where the microbiome is fully described, or ‘specific pathogen free’ (SPF) animals where a moderately normal microbiome is present but devoid of pathogenic microorganisms. Pioneering GF research in poultry in the late 1940s and 1950s has its origin in a need understand the mode of action of antibiotics. Early researchers quickly established that GF chicks responded differently to antibiotics than CV counterparts. The GF experimental model has since been exploited in many divergent fields including pathology, immunology, metabolism, anatomy, physiology, and others. The absence of a microbiome presents the host with a range of advantages and disadvantages. For example, GF chicks often grow more quickly and have lower feed conversion ratio (FCR) than their CV counterparts but may be less resilient to external stress and have a compromised immunological maturation rate. This review will summarize the literature on GF animal research with a special emphasis on poultry. The objective of the review is to establish a frame of reference to understand the extent of the role of the microbiome in animal health, welfare, nutrition, and growth, to provide opportunities for targeted modulation of the microbiome to achieve desired phenotypic responses whilst simultaneously minimizing unintended collateral effects.
Diets fed to nonruminant animals are composed mainly of feed ingredients of plant origin. A variety of antinutritional factors such as phytin, nonstarch polysaccharides, and protease inhibitors may ...be present in these feed ingredients, which could limit nutrients that may be utilized by animals fed such diets. The primary nutrient utilization-limiting effect of phytin arises from the binding of 6 phosphate groups, thus making the P unavailable to the animal. The negative charges allow for formation of insoluble phytin-metal complexes with many divalent cations. Furthermore, phytin and protein can form binary complexes through electrostatic links of its charged phosphate groups with either the free amino group on AA on proteins or via formation of ternary complexes of phytin, Ca2+, and protein. The form and extent of de novo formation of binary and ternary complexes of phytin and protein are likely to be important variables that influence the effectiveness of nutrient hydrolysis in plant-based diets. Nonstarch polysacharides reduce effective energy and nutrient utilization by nonruminant animals because of a lack of the enzymes needed for breaking down the complex cell wall structure that encapsulate other nutrients. Enzymes are used in nonruminant animal production to promote growth and efficiency of nutrient utilization and reduce nutrient excretion. The enzymes used include those that target phytin and nonstarch polysaccharides. Phytase improves growth and enhances P utilization, but positive effects on other nutrients are not always observed. Nonstarch polysaccharide-hydrolyzing enzymes are less consistent in their effects on growth and nutrient utilization, although they show promise and it is imperative to closely match both types and amounts of nonstarch polysaccharides with appropriate enzyme for beneficial effects. When used together with phytase, nonstarch polysaccharide-hydrolyzing enzymes may increase the accessibility of phytase to phytin encapsulated in cell walls. The future of enzymes in nonruminant animal production is promising and will likely include an understanding of the role of enzyme supplementation in promoting health as well as how enzymes may modulate gene functions. This review is an attempt to summarize current thinking in this area, provide some clarity in nomenclature and mechanisms, and suggest opportunities for expanded exploitation of this unique biotechnology.
A total of 360 Ross 708 male broiler chicks were used in an 8 treatment and 9 replicate cage study to explore the influence of day-old chick weight on the efficacy of exogenous phytase. Treatments ...were arranged as a 2 × 4 factorial with the factors being diet (a positive (PC) and negative control (NC) varying in nutrient density fed without or with 2 concentrations of exogenous phytase) and chick weight (light; <38.5 g or heavy; >42 g). Chicks were sourced from the same breeder flock, with light and heavy chicks being selected from the naturally occurring heterogeneity in the population. The diets were corn-soybean meal based and the PC was formulated to meet the nutrient requirements of male broiler chicks. The NC was formulated to contain 120 kcal/kg, 0.5, 0.18, and 0.18% less apparent metabolizable energy (AME), crude protein (CP), calcium (Ca), and phosphorus (P), respectively, than the PC. Amino acid (AA) density in the NC was also reduced in line with the reduction in CP and the manufacturers’ recommendations for the effect of phytase on amino acid digestibility. Phytase at either 1,000 FYT/kg or 3,000 FYT/kg was added only to the NC to create a total of 4 diets. Water and mash feed were available ad libitum and were offered to 8 replicate cages, each containing 5 chicks. The experiment was conducted over a period of 3 wk and diets were offered in 2 phases (starter from d 1 to 10 and grower from d 11 to 21). Growth performance was monitored at the end of each diet phase and on d 21 ileal digesta were collected for estimation of apparent digestibility of energy (DE), nitrogen (N), Ca, P, dry matter (DM), and AA. There were no statistically significant interactions between diet and day-old chick weight for any of the measured parameters. Light chicks had significantly lower weight gain (approx. 5%) at both d 10 and d 21 compared with heavy chicks. This effect was principally associated with reduced feed intake and there was no significant effect of chick weight on feed conversion ratio (FCR). Chick weight had no effect on ileal nutrient digestibility. The reduction in nutrient density from the PC to the NC generated a significant reduction in weight gain (around 12%) and a significant increase in FCR (1.68 vs. 1.83). This effect was associated with a significant reduction in ileal DE (approximately 150 kcal/kg) and in the digestibility of several AA. Exogenous phytase significantly increased weight gain, reduced FCR and generated a significant increase in the ileal digestibility of energy, N, P, and several AA. Although chick weight and diet did not interact statistically, heavy chicks benefited more than light chicks from high doses of exogenous phytase across almost all measured end points which was confirmed by regression analysis. In conclusion, light chicks have inferior performance outcomes than heavy chicks principally because of reduced feed intake, but putatively not in digestive capacity per se. Exogenous phytase is effective in improving performance and nutrient digestibility in nutrient deficient diets. The effect of chick weight per se, and also breeder flock age, on the utility of supra-nutritional inclusion concentrations of exogenous phytase warrants further study.
The use of enzymes has increased dramatically in the last 20years, and yet the fact that perhaps one of their most important actions is in mediating the size and composition of microbial populations ...is still largely unrecognized. As the public press for increasingly antibiotic (and by extension coccidiostat) free diets, it is clear that this under-investigated facet of enzyme action warrants far greater scrutiny. Until quite recently it was assumed that the intestinal microbiota were simply reactive to the nutrients left behind after digestion had completed. However it now seems that intestinal microbial population size and composition clearly plays a very large role in determining the extent of digestion accomplished by the host and by extension, growth rate and efficiency. This is becoming increasingly becoming apparent with the advent of new molecular techniques coupled with statistical methods which are beginning to identify desirable and undesirable clusters of organisms as far as good performance is concerned, although the variability in techniques utilised is hampering progress. The fact that exogenous enzymes not only influence the partitioning of nutrients to the host but also, through their action, produce nutrients for specific populations of bacteria, means that they are multifactorial in their effect. As a result their effects are influenced by many other factors such as presence of antibiotics, health status/disease challenge of the animals and quality of ingredients fed. Understanding of such interactions will lead to a more predictable and profitable outcome for the end user.
The effects of phytic acid and microbial phytase on the flow and composition of endogenous protein at the terminal ileum of broiler chickens were investigated using the peptide alimentation method. ...Phytic acid (fed as the sodium salt) was included in a synthetic diet at 8.5, 11.5 and 14.5 g/kg (or 2.4, 3.2 and 4.0 g/kg phytate-phosphorus) and each diet was fed without or with an Escherichia coli-derived microbial phytase at 500 phytase units/kg diet. A control containing no phytate was fed as a comparison to estimate basal endogenous flows. Ingestion of phytic acid increased (P<0.05) the flow of endogenous amino acids and N by an average of 47 % at the lowest phytic acid concentration and 87 % at the highest. The addition of microbial phytase reduced (P<0.05) the inimical effects of phytic acid on endogenous amino acid flow at all dietary phytic acid levels. The composition of endogenous protein was also influenced (P< 0.10-0.001) by increasing phytic acid concentrations and phytase addition. The effects of phytic acid and phytase on endogenous flow and composition of endogenous protein, however, varied depending on the amino acid. It is concluded that the effects of phytase on amino acid digestibility may be mediated, in part, through a route of reduced endogenous loss.
A total of 480 male Cobb 500 broiler chicks were assigned to one of 6 dietary treatments to explore the energy equivalence of myo-inositol compared with dextrose. The 6 dietary treatments included a ...corn and soy-based control ration formulated with 5% anhydrous dextrose and 5 further diets that were generated by the sequential displacement of increments of 1% dextrose with myo-inositol. Each diet was fed to 8 replicate cages of 10 chicks per cage from day 8 to day 18 after hatch. The BW gain, feed intake, and feed conversion ratio (FCR) were measured, and on day 15 to day 17, excreta were collected to estimate the total tract nutrient retention. Ileal digestibility of nutrients and tibia mineral content was assessed on day 18. The displacement of dextrose with myo-inositol generated a significant linear reduction in the FCR that did not reach a plateau at 5% dietary inclusion of myo-inositol. There was no effect of the displacement of dextrose with myo-inositol on bone mineral concentration. However, supplemental myo-inositol linearly reduced ileal digestibility of DM, calcium, and ileal digestible energy. Myo-inositol addition resulted in a significant linear increase in the total tract retention of CP. It can be concluded that myo-inositol has an energy equivalence equal to approximately 78% of that of dextrose for young broiler chicks but exerts a range of extra caloric effects that improve feed efficiency and may influence nitrogen (N) retention and the uric acid cycle. Future work should focus on the role of phytase and myo-inositol on uric acid, creatine kinase, and other metabolites involved in renal function and biochemical flows of N in urine and feces in nonruminants.
Trypsin inhibitors (TI) resident in soybeans affects protein utilization. While heat treatment influences residual TI, it simultaneously affects the structure and solubility of the soybean proteins ...and confounds any response to exogenous proteases. Using purified TI, the effect of exogenous protease to TI can be dissociated from changes in the soybean protein. Thus, the current study was designed to evaluate the growth performance and protein utilization responses of broiler chickens to purified TI and exogenous protease. Soybean meal (SBM) was preanalyzed for basal TI (2,996 TIU/g SBM), formulated into nutritionally adequate experimental diets to contain 1,033 TIU/g diet, and purified TI was added at 9,000 TIU/g diet. A total of 320 Cobb-500 broiler chicks were allocated to 4 diets, each with 8 replicate cages and 10 birds per replicate. The experimental diets were arranged as a 2 × 2 factorial with factors being dietary TI (1,033 or 10,033 TIU/g) and exogenous protease (0 or 15,000 PROT/kg). On day 7, 14, and 21 posthatching, protease supplementation improved the BW gain (P < 0.01) and gain to feed ratio (P < 0.05) of birds. On day 14 and 21 posthatching, the relative weight of pancreas increased (P < 0.05) with added TI but was reduced (P < 0.001) with protease supplementation. Apparent ileal digestibility of all amino acids, except methionine, decreased (P < 0.001) with added TI but increased (P < 0.05) with protease supplementation. Jejunal MUC-2 was downregulated (P < 0.01) and SCL7A-2 was upregulated (P < 0.05) by protease supplementation. Duodenal trypsin and chymotrypsin activities reduced (P < 0.05) with added TI but increased (P < 0.01) with protease supplementation. Exogenous protease produced longer villi (P < 0.05) and deeper crypts (P < 0.01) in the jejunal tissue. In conclusion, dietary addition of purified TI negatively affects nutrient utilization by broiler chickens. Furthermore, the study showed that the efficacy of the exogenous protease might be independent of dietary TI concentration.
Phytases have been used commercially since the early 1990s and have been the focus of considerable and sustained research for many decades. Despite this heroic effort there are still areas of ...persistent uncertainty such as the obscurity surrounding total compared with digestible calcium, appropriate modification to dietary sodium (and other electrolyte) concentrations, the usefulness of the amino acid and energy digestibility improvements and ultimately the effect of phytase on nutrient requirement. One further area which has attracted some attention recently is the effect of unconventionally high doses of phytase (i.e. >2,500 FTU/kg from Aspergillus niger or Escherichia coli) in an attempt to ostensibly 'de-phytinise' the diet. The effects of such 'super' doses of phytase can be considerable, and often beyond that which may be reasonably expected based on improvement in P digestibility per se. This review article addresses these effects and suggests mechanisms by which they may be explained.
A total of 945 male Ross 308 broiler chicks were used in a growth study to explore the interaction between dietary crude protein concentration and available phosphorus. Nine experimental treatments ...were constructed factorially by offering low, medium, or standard protein concentrations without or with low, standard, or high available phosphorus. Diets were based on corn, wheat, and soybean meal and all nutrients other than protein/amino acids and available phosphorus were maintained at or above breeder guidelines. Additional synthetic amino acids were used in the diets with low protein concentration in attempt to maintain digestible amino acid supply. Diets were offered to 7 replicate pens of 15 chicks per pen from day 8 to 35. Growth performance was measured during the grower (day 8–24) and finisher (day 25–35) periods. On day 35 carcass composition was determined, blood was drawn for various biochemical measurements and the tibia was excised for mechanical and compositional analyses. Birds that received the low-protein diet had lower terminal body weight and higher feed conversion ratio compared with those that received diets with adequate crude protein content. However, addition of available phosphorus to the low-protein diet resulted in significant reductions in weight-corrected feed conversion that were not evident in the diet with adequate protein content. Bone architecture was only moderately influenced by dietary treatment but birds that ingested the diets containing low and medium protein concentrations had relatively heavier abdominal fat pad weight. Blood biochemistry, especially ammonia, uric acid, and phosphorus, was influenced by both dietary protein and available phosphorus and trends suggested that both axes are involved in protein accretion and catabolism. It can be concluded that performance losses associated with feeding low protein diets to broiler chickens may be partially restored by additional available phosphorus. The implications for use of exogenous enzymes such as protease and phytase and protein nutrition per se warrants further examination.
A total of 1,408 male broiler chickens were used to evaluate the impact of age and duration of feeding low-phosphorus (P) diet on the efficacy of phytase using growth performance, nutrient ...utilization, tibia ash, and plasma indices. Diets were formulated with 2 non-phytate P (nPP) concentrations (i.e., 0.20 and 0.40%) and 2 phytase concentrations (i.e., 1,000 and 2,000 FTU/kg) added to the 0.20% nPP diet. Four dietary treatments with 8 replicate cages each were fed to broiler chicks at different ages and for different duration. Specifically, these were days 6 to 8 (12 birds per replicate), 12 to 14, 9 to 14, 20 to 22, or 6 to 22 (8 birds per replicate). Parameters were measured on the last day of each period. Duration of feeding effect was examined by comparing responses of birds fed for 2 or 5 D at day 14 and for 2 or 16 D at day 22; age effect was determined by comparing responses of birds fed for 2 D at age 8, 14 and 22 D post-hatching. Body weight gain and gain-to-feed ratio were increased (P < 0.01) in birds fed diets supplemented with phytase; however, an increase in duration of feeding improved (P < 0.01) feed efficiency with birds fed for 16 D performing better than birds fed for 2 D at day 22. In addition, phytase supplementation improved (P < 0.01) apparent P and calcium digestibility and retention, but the age effect on phytase efficacy was more apparent at day 14 and the duration of feeding effect was evident in birds fed for 2 D due to the increased levels of mineral utilization at that age/duration of feeding as compared with the other groups. The results of this study show that phytase efficacy was at optimum in birds fed low-P diet for 2 D at day 14. This period can be recommended for further bioefficacy studies of phytase.
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