Shane M. Rutherfurd

The effect of microbial phytase on ileal phosphorus and amino acid digestibility in the broiler chicken

1. The study aimed to assess the effect of a commercially available microbial phytase on phytate phosphorus and total phosphorus content at the terminal ileum as well as true ileal amino acid digestibility. 2. Five diets, each containing a different plant-based feedstuff, were supplemented with microbial phytase and fed, along with a non-supplemented corresponding diet, to 28-d-old broiler chickens, Chromic oxide was used as an indigestible marker. Ileal contents were collected and analysed, along with the diets, for total phosphorus, phytate phosphorus and amino acids. 3. Endogenous phosphorus determined at the terminal ileum was 272 ± 108 mg/kg food dry matter (mean ± SE). Endogenous ileal amino acid flows ranged from 58 ± 10 mg/kg food dry matter for methionine to 568 ± 47 mg/kg food dry matter for glutamic acid. 4. Supplementation with microbial phytase resulted in a significantly greater phytate P disappearance from the terminal ileum for rice bran (17% units), but not for soyabean meal, maize, wheat or rapeseed meal. Similarly total phosphorus digestibility was significantly ( P < 0.05) higher when microbial phytase was added to the rice-bran-based diet but not for any of the other feedstuffs. 5. Amino acid digestibility was significantly greater in the presence of microbial phytase for all the amino acids examined in wheat, for several of the amino acids each in maize and rapeseed meal and for one amino acid in rice bran and soyabean meal. The average increase in amino acid digestibility for those amino acids affected, was 13, 6, 10, 7 and 12% units for wheat, maize, rapeseed meal, rice bran and soyabean meal, respectively. 6. It appears that microbial phytase improves phosphorus digestibility and amino acid digestibility for certain plant-based feedstuffs.

Protein Digestibility-Corrected Amino Acid Scores and Digestible Indispensable Amino Acid Scores Differentially Describe Protein Quality in Growing Male Rats

BACKGROUND The FAO has recommended replacing the protein digestibility-corrected amino acid score (PDCAAS) with the digestible indispensable amino acid score (DIAAS). OBJECTIVE The objective of this study was to compare aspects underlying the calculation of the DIAAS and PDCAAS, including 1) fecal digestibility vs. ileal digestibility, 2) using a single nitrogen digestibility value for all amino acids, and 3) the effect of truncation. Truncated PDCAAS and untruncated DIAAS values calculated as formally defined were also compared and DIAAS data presented for 14 dietary protein sources. METHODS Semisynthetic wheat starch-based diets were formulated to contain the test protein (as consumed by humans) source (whey- and soy-protein isolates, milk-, whey-, rice- and pea- protein concentrates, cooked kidney beans, roasted peanuts, cooked peas, corn-based breakfast cereal, cooked rice, cooked rolled oats, and wheat bran) as the sole nitrogen source and with an indigestible marker (titanium dioxide). Growing male rats (∼250 g bodyweight) were given a basal casein-based diet from day 1 to day 7 and then allocated (n = 6) to the test diets for day 8 to day 14 before ileal digesta were collected after the rats were killed. Total feces were collected from day 11 to day 14. RESULTS True fecal nitrogen digestibility was different (P < 0.05; 10% difference on average) from true ileal nitrogen digestibility for 11 of the 14 protein sources. True ileal nitrogen digestibility was different (P < 0.05) from true ileal amino acid digestibility for almost half of the indispensable and conditionally indispensable amino acids (differences ranged from 0.9% to 400%). DIAAS values ranged from 0.01 for a corn-based cereal to 1.18 for milk protein concentrate. CONCLUSION Untruncated PDCAAS values were generally higher than a DIAAS values, especially for the poorer quality proteins; therefore, the reported differences in the scores are of potential practical importance for populations in which dietary protein intake may be marginal.

In vivo digestion of bovine milk fat globules: Effect of processing and interfacial structural changes. I. Gastric digestion

The aim was to study the in vivo gastric digestion of fat globules in bovine cream from raw, pasteurised or pasteurised and homogenised milk. Fasted rats were gavaged once and chyme samples were collected after 30, 120 and 180 min post-gavage. Proteins from raw (RC) and pasteurised (PC) creams appeared to be digested faster and to a greater extent. Free fatty acids (FAs) increased throughout the 3h postprandial period. Short and medium chain FAs were released more rapidly than long chain FAs which were hydrolysed to a greater degree from PC. The size of the fat globules of all creams increased in the stomach. Protein aggregates were observed in pasteurised and homogenised cream chyme. Protrusions, probably caused by the accumulation of insoluble lipolytic products, appeared at the surface of the globules in RC and PC chyme. Overall, PC proteins and lipids appeared to be digested to a greater extent.

Actinidin Enhances Gastric Protein Digestion As Assessed Using an in Vitro Gastric Digestion Model

Consumption of kiwifruit has long been claimed anecdotally to assist in gastric digestion. This has generally been assumed to be due to the presence of the proteolytic enzyme actinidin; however, there is little published evidence supporting this assumption. This paper reports the findings of an in vitro study that examined the effect of kiwifruit proteases (actinidin) on the digestion of a range of common food proteins under simulated gastric conditions. An extract from green kiwifruit containing actinidin was prepared. Several protein sources derived from soy, meat, milk, and cereals were incubated in the presence or absence of the kiwifruit extract using an in vitro digestion system consisting of incubation with pepsin at pH 1.9, simulating gastric digestion in humans. The digests were subjected to gel electrophoresis (SDS-PAGE). For some protein sources, simulated digestion in the presence of kiwifruit extract resulted in a substantially greater loss of intact protein and different peptide patterns from those seen after digestion with pepsin alone. As an example, the addition of actinidin extract enhanced the digestion of alpha-, beta-, and kappa-caseins in sodium caseinate by 37, 33, and 48%, respectively. Under simulated gastric conditions, kiwifruit extract containing actinidin enhanced the digestion of some, but not all, food proteins over and above that found with pepsin alone.

Actinidin Enhances Protein Digestion in the Small Intestine As Assessed Using an in Vitro Digestion Model

This paper describes an in vitro study that tests the proposition that actinidin from green kiwifruit influences the digestion of proteins in the small intestine. Different food proteins, from sources including soy, meat, milk, and cereals, were incubated in the presence or absence of green kiwifruit extract (containing actinidin) using a two-stage in vitro digestion system consisting of an incubation with pepsin at stomach pH (simulating gastric digestion) and then with added pancreatin at small intestinal pH, simulating upper tract digestion in humans. The digests from the small intestinal stage (following the gastric digestion phase) were subjected to gel electrophoresis (SDS-PAGE) to assess loss of intact protein and development of large peptides during the in vitro simulated digestion. Kiwifruit extract influenced the digestion patterns of all of the proteins to various extents. For some proteins, actinidin had little impact on digestion. However, for other proteins, the presence of kiwifruit extract resulted in a substantially greater loss of intact protein and different peptide patterns from those seen after digestion with pepsin and pancreatin alone. In particular, enhanced digestion of whey protein isolate, zein, gluten, and gliadin was observed. In addition, reverse-phase HPLC (RP-HPLC) analysis showed that a 2.5 h incubation of sodium caseinate with kiwifruit extract alone resulted in approximately 45% loss of intact protein.

Effect of a novel phytase on growth performance, apparent metabolizable energy, and the availability of minerals and amino acids in a low-phosphorus corn-soybean meal diet for broilers

The addition of microbial phytase to diets for broiler chickens has been shown to improve the availability of phytate P, total P, some other minerals, and amino acids. In this study, the effect of a novel microbial phytase expressed by synthetic genes in Aspergillus oryzae on amino acid and mineral availability was assessed. Phytase was incorporated (1,000 and 2,000 U/kg) into low-P corn-soybean meal-based diets for broilers. Broilers received the experimental diets for 3 wk, and excreta were collected from d 18 to 21 for the determination of AME and mineral retention. On the 22nd day, the broilers were killed and the left leg removed and ileal digesta collected. Ileal phytate P and total P absorption, ileal amino acid digestibility, as well as the bone mineral content and bone mineral density were determined. Ileal phytate P absorption and absorbed phytate P content of the low-P corn-soybean meal diet were significantly (P < 0.05) higher after dietary inclusion of the novel phytase (49-60% and 65-77% higher, respectively). Apparent ileal total P absorption and apparent total P retention was 12 to 16% and 14 to 19% higher (P < 0.05), respectively, after dietary inclusion of phytase. The bone mineral content and bone mineral density in the tibia were 32 to 35% and 19 to 21% higher (P < 0.05), respectively, after dietary phytase inclusion. The apparent ileal digestibility of threonine, tyrosine, and histidine increased significantly (P < 0.05) by 14, 9, and 7%, respectively, after dietary inclusion of microbial phytase. Overall, the inclusion of a novel microbial phytase into a low-P corn-soybean meal diet for broiler chickens greatly increased phytate P and total P absorption, bone mineral content and density, as well as the digestibility of some amino acids.

Endogenous flow of amino acids in the avian ileum as influenced by increasing dietary peptide concentrations

The aim of the present study was to establish whether feeding broiler chickens with diets containing increasing dietary peptide concentrations would cause increases in ileal endogenous amino acid flow. The flow of N and most amino acids increased quadratically (P < 0.05 to 0.001) with increasing dietary concentrations of peptides. The exceptions were the flow of threonine, serine, glycine, tyrosine and cystine, which increased linearly (P < 0.001) with dietary peptide levels. Another notable exception to the general trend was the flow of proline, which was significantly higher (P < 0.01) in birds fed the protein-free diet. The amino acid profile of endogenous protein, expressed as proportion of crude protein, indicated that the ratios of threonine, glutamic acid, proline, glycine, leucine, histidine, arginine and cystine were influenced (P < 0.05) with increasing dietary peptide concentrations. In general, compared with the protein-free diet, the ratios of threonine and arginine in endogenous protein were lower (P < 0.05) and those of glutamic acid, glycine and histidine were greater (P < 0.05) in diets with high concentrations of peptides. The ratio of proline was found to decrease (P < 0.05) with increasing dietary peptide concentrations. These changes in the amino acid profile of endogenous protein are probably reflective of changes in the output of one or more of the components of endogenous protein. Overall, the present results demonstrated that increasing dietary peptide concentrations increased the flow of endogenous amino acid flow at the terminal ileum of broiler chickens in a dose-dependent manner and also caused changes in the composition of endogenous protein. The observed changes in endogenous amino flow will influence the maintenance requirements for amino acids and also have implications for the calculation of true digestibility coefficient of feedstuffs.

The extent of ovalbumin in vitro digestion and the nature of generated peptides are modulated by the morphology of protein aggregates

The impact of heat-induced aggregation on the extent of ovalbumin digestion and the nature of peptides released was investigated using an in vitro digestion model. The extent of hydrolysis, estimated by the disappearance of intact ovalbumin and the appearance of soluble peptides, was greater for the linear aggregates as compared to the spherical aggregates. The latter result may be due to differences in the surface area to volume ratio of the aggregates, or the degree of unfolding of the proteins during aggregate preparation. Peptide identification using LC-MS/MS highlighted that ovalbumin aggregation rendered a number of peptide bonds accessible to digestive proteases which were not accessible in native ovalbumin. Moreover, the peptide bonds that were cleaved appeared to be specific depending on the morphology of the aggregates. This work illustrates the links existing between food structure and their breakdown during the digestive process. Such quantitative and qualitative differences may have important nutritional consequences.

Development of a novel bioassay for determining the available lysine contents of foods and feedstuffs

Lysine is an important indispensable amino acid, and describing the lysine content of a food or feedstuff provides useful information about nutritional value. However, when a food or feedstuff is subjected to heating the lysine present can be altered to nutritionally unavailable derivatives. These derivatives can revert back to lysine during the acid hydrolysis step used in amino acid analysis causing an overestimate of the lysine content. There have been many chemical methods developed to determine the reactive (unmodified) lysine content of foods and feedstuffs, but these do not take into account the incomplete absorption of lysine from the small intestine. There are also a number of animal-based assays for determining available lysine (the lysine that can be absorbed in a form that can be used for protein synthesis). The true ileal amino acid digestibility assay is commonly used to determine amino acid availability and is accurate for application to unprocessed foods and feedstuffs but is not accurate for lysine and possibly other amino acids when applied to heat-processed foods or feedstuffs. For such protein sources, assays such as the slope-ratio assay, indicator amino acid oxidation assay and the BIOLYSINEtrade mark assay (true ileal digestible reactive lysine assay) have been developed to determine available lysine. The present paper discusses the efficacy of the BIOLYSINEtrade mark assay as well as other assays for determining available lysine in processed foods and feedstuffs.

In vivo digestion of bovine milk fat globules: effect of processing and interfacial structural changes. II. Upper digestive tract digestion.

The aim of this research was to study the effect of milk processing on the in vivo upper digestive tract digestion of milk fat globules. Fasted rats were serially gavaged over a 5h period with cream from raw, pasteurised, or pasteurised and homogenised milk. Only a few intact dietary proteins and peptides were present in the small intestinal digesta. Significantly (P<0.05) more longer chain (C≥10) fatty acids were present in the digesta of rats gavaged with raw (448 mg g(-1) digesta dry matter (DDM)) and homogenised creams (528 mg g(-1) DDM), as compared to pasteurised and homogenised cream (249 mg g(-1) DDM). Microscopy techniques were used to investigate the structural changes during digestion. Liquid-crystalline lamellar phases surrounding the fat globules, fatty acid soap crystals and lipid-mucin interactions were evident in all small intestinal digesta. Overall, the pasteurised and homogenised cream appeared to be digested to a greater extent.