Peter J. Royle

Whey proteins as functional food ingredients

Putative anti-cancer activity of whey proteins has been investigated in an animal model to evaluate their potential role in disease prevention, and to contribute to a basis for their inclusion as ingredients in functional foods. Animal feeding trials have compared the efficacy of dietary whey proteins in retarding chemically induced colon cancer in a rat model of the disease. Dairy proteins, in particular whey protein, were found to be efficacious in retardation of colon cancer in young rats compared with other dietary proteins (meat, soy). The influence of dietary whey protein on development of colon cancer in mature rats has also been examined. Results similar to those with younger animals have been demonstrated, a finding that suggests age does not significantly alter the outcome. Efficacy of whey protein fractions has also been assessed. Preliminary results suggest that diets supplemented with lactoferrin or with β-lactoglobulin enhance protection against the development of putative tumor precursors (aberrant crypts) in the hind gut wall. The mechanism behind the apparent anti-cancer activity of dietary whey proteins in these studies may be related to their sulfur amino acid content, for which there is a high requirement in the rat, and hypothesised role in protecting DNA in methylated form. In a parallel study, a number of potential functional foods containing whey protein (flavored milk, pasta, ice cream, dessert pudding, muesli, and savory dip) have been developed in preparation for human clinical trials. The foods containing whey protein were generally highly acceptable in taste trials. These products are expected to be suitable as delivery vehicles for dietary whey protein in studies aimed at substantiating the human health benefits of this protein source.

A Probiotic Strain of L. Acidophilus Reduces DMH-Induced Large Intestinal Tumors in Male Sprague-Dawley Rats

Probiotic bacteria strains were examined for their influence on 1,2-dimethylhydrazine (DMH)-induced intestinal tumors in 100 male Sprague-Dawley rats. Lactobacillus acidophilus (Delvo Pro LA-1), Lactobacillus rhamnosus (GG), Bifidobacterium animalis (CSCC1941), and Streptococcus thermophilus (DD145) strains were examined for their influence when added as freeze-dried bacteria to an experimental diet based on a high-fat semipurified (AIN-93) rodent diet. Four bacterial treatments were compared: L. acidophilus, L. acidophilus + B. animalis, L. rhamnosus, and S. thermophilus, the bacteria being added daily at 1% freeze-dried weight (10(10) colony-forming units/g) to the diet. Trends were observed in the incidence of rats with large intestinal tumors for three treatments: 25% lower than control for L. acidophilus, 20% lower for L. acidophilus + B. animalis and L. rhamnosus treatments, and 10% lower for S. thermophilus. Large intestinal tumor burden was significantly lower for treated rats with L. acidophilus than for the control group (10 and 3 tumors/treatment group, respectively, p = 0.05). Large intestinal tumor mass index was also lower for the L. acidophilus treatment than for control (1.70 and 0.10, respectively, p < 0.05). Other treatments showed no statistically significant change from control for these indexes of tumorigenesis. For rats fed L. acidophilus, no adenocarcinomas were present in the colons. Pulsed-field gel electrophoresis of bacterial chromosomal DNA fragments was used to differentiate introduced (exogenous) bacterial strains from indigenous bacteria of the same genera present in the feces. Survival during gut passage and displacement of indigenous lactobacilli occurred with introduced L. acidophilus and L. rhamnosus GG during the probiotic treatment period. However, introduced strains of B. animalis and S. thermophilus were not able to be isolated from feces. It is concluded that this strain of L. acidophilus supplied as freeze-dried bacteria in the diet was protective, as seen by a small but significant inhibition of tumors within the rat colon.

Whey protein isolate and glycomacropeptide decrease weight gain and alter body composition in male Wistar rats

The effect of feed protein type on body composition and growth has been examined. Evidence exists that whey protein concentrate is effective at limiting body fat expansion. The presence of caseinomacropeptide, a mixture of glycosylated and non-glycosylated carbohydrate residues, in particular glycomacropeptide (GMP) in whey protein concentrate may be important for this effect. The influence of whey protein isolate (WPI) and GMP on weight gain and body composition was examined by feeding Wistar rats ad libitum for 7 weeks with five semi-purified American Institute of Nutrition-based diets differing in protein type: (1) casein; (2) barbequed beef; (3) control WPI (no GMP); (4) WPI+GMP at 100 g/kg; (5) WPI+GMP at 200 g/kg. Body composition was assessed, and plasma samples were assayed for TAG, insulin and glucose. Body-weight gain was lower (- 21 %) on the control WPI diet relative to casein, with a non-significant influence associated with GMP inclusion (- 30 %), the effect being equivalent at both levels of GMP addition. Renal and carcass fat mass were reduced in the highest GMP diet when compared with WPI (P < 0.05). Plasma insulin was lowered by GMP at the highest addition compared with WPI alone (- 53 %; P < 0.01). Plasma TAG in the WPI+GMP (200 g/kg) group were lower (- 27 %; P < 0.05) than the casein and beef groups. In conclusion, GMP appears to have a significant additional influence when combined with WPI on fat accumulation. WPI alone appears to have the predominant influence accounting for 70 % of the overall effect on body-weight gain. Mechanisms for this effect have not been identified but food intake was not responsible.

A comparative study of the influence of differing barley brans on DMH-induced intestinal tumours in male Sprague-Dawley rats.

The influence of barley brans on the incidence and burden of intestinal tumours in rats induced by 1,2‐dimethylhydrazine (DMH) was studied in a 7 month feeding experiment. The basic diet was American Institute of Nutrition (AIN) 76 modified by adjustment to 20% fat and 40% starch; brans were added so as to supply 5% dietary fibre. The barley brans studied were commercial barley bran (BB1; 13.0% dietary fibre) from the aleurone/subaleurone layer, outerlayer barley bran (BB2) including the germ (25.5% dietary fibre) and spent barley grain bran (SBG; a by‐product of the brewery and including the hull; 47.7% dietary fibre). They were compared with wheat bran (WB; 44% dietary fibre) and cellulose (or control; 98% dietary fibre). Commercial barley bran and wheat bran were most effective in reducing tumour incidence and burden. The incidence of tumours fell significantly from 70% (BB2) and 50% (SBG) to 10% (BB1) and 20% (WB) and tumour burden and tumour mass index (TMI) were also reduced by similar orders of magnitude. There were significantly higher short chain fatty acid (SCFA) concentrations in WB‐ and BBl‐fed rat faecal pellets relative to cellulose‐ and BB2‐fed rat faeces; butyrate, in particular, was affected. Regression analysis of butyrate against tumour incidence showed a trend (r=‐0.898; P=0.055), but the concentration of butyrate alone could not account for the reduction in tumour incidence observed. In a second experiment, when two brans (BB1 and SBG) were introduced after DMH dosing, there were higher incidences and burdens of tumours, indicating that protection by such brans was not as effective under these circumstances. Commercially available barley bran and wheat bran appear to significantly reduce tumour incidence and burden in this model relative to other brans, influencing both the initiatory as well as promotional stages of chemically induced carcinogenesis.

Bioequivalence of n -3 fatty acids from microencapsulated fish oil formulations in human subjects

Fish oil n-3 fatty acids (FA) have known health benefits. Microencapsulation stabilises and protects fish oil from oxidation, enabling its incorporation into foods. The aim of the present study was to compare the bioavailability of n-3 FA delivered as two microencapsulated fish oil-formulated powders or fish oil gel capsules (FOGC) taken with a flavoured milk in healthy participants. Formulation 1 (F1) composed of a heated mixture of milk protein-sugar as an encapsulant, and formulation 2 (F2) comprised a heated mixture of milk protein-sugar-resistant starch as an encapsulant. Participants consumed 4 g fish oil (approximately 1·0 g EPA and DHA equivalent per dose). Bioavailability was assessed acutely after ingestion of a single dose by measuring total plasma FA composition over a period of 48 h (n 14) using a randomised cross-over design, and over the short term for a period of 4 weeks using an unblinded parallel design (after daily supplementation) by measuring total plasma and erythrocyte FA composition at baseline and at 2 and 4 weeks (n 47). In the acute study, F1 greatly increased (% Δ) plasma EPA and total n-3 FA levels at 2 and 4 h and DHA levels at 4 h compared with FOGC. The time to reach maximal plasma values (T(max)) was shorter for F1 than for FOGC or F2. In the short-term study, increases in plasma and erythrocyte n-3 FA values were similar for all treatments and achieved an omega-3 index in the range of 5·8-6·3 % after 4 weeks. Overall, the results demonstrated human bioequivalence for microencapsulated fish oil powder compared with FOGC.