The affinity of milk fat globule membrane fragments and buttermilk proteins to hydroxyapatite.

Buttermilk differs from skim milk by the presence of milk fat globule membrane (MFGM) fragments that are released during cream churning. MFGM is rich in health-promoting components, such as phospholipids and membrane proteins, but these compounds have a negative impact on buttermilk techno-functional properties in dairy applications. The isolation of MFGM from buttermilk improved its functionality while also recovering the MFGM bioactive components. Hydroxyapatite (HA) can be used to extract MFGM by adsorption via charged site interactions. However, the affinity of HA to MFGM or the main buttermilk proteins (casein micelles (CM), ß-lactoglobulin (ß-lg) and α-lactalbumin (α-lac)) is not known. The influence of important physicochemical parameters such as pH and temperature on these interactions is also unclear. For each buttermilk component, a quartz crystal microbalance diffusion analysis was performed to determine the maximum adsorption time and the attached mass density on HA-coated gold sensors. The influence of pH, ionic strength (IS), and temperature (T) on the affinity of each buttermilk component for HA particles was assessed using a 3-levels and 3-factors Box-Behnken design. The absorption rate was highest for the CM, followed by ß-lg and α-lac, and then by the MFGM. Nevertheless, the final maximal attached mass densities to the HA were similar for the MFGM and CM, and 2.5 times higher than for ß-lg and α-lac. This difference can be explained by the higher number of binding sites found in CM and their heavier mass. The model obtained by the Box-Behnken design plan showed that the adsorption of the CM changed with T, pH and IS. These results suggest that the techno-functional properties of buttermilk may be restored by specifically extracting MFGM with HA. Experiments are ongoing to determine conditions for fractionating MFGM directly from buttermilk.

Effect of reverse osmosis and ultra-high-pressure homogenization on the composition and microstructure of sweet buttermilk.

Buttermilk (BM), the by-product of butter making, is similar to skim milk (SM) composition. However, it is currently undervalued in dairy processing because it is responsible for texture defects (e.g., crumbliness, decreased firmness) in cheese and yogurt. One possible way of improving the incorporation of BM into dairy products is by the use of technological pretreatments such as membrane filtration and homogenization. The study aimed at characterizing the effect of preconcentration by reverse osmosis (RO) and single-pass ultra-high-pressure homogenization (UHPH) on the composition and microstructure of sweet BM to modify its techno-functional properties (e.g., protein gel formation, syneresis, firmness). The BM and RO BM were treated at 0, 15, 150, and 300 MPa. Pressure-treated and control BM and RO BM were ultracentrifuged to fractionate them into the following 3 fractions: a supernatant soluble fraction (top layer), a colloidal fraction consisting of a cloudy layer (middle layer), and a high-density pellet (bottom layer). Compositional changes in the soluble fraction [lipid, phospholipid (PL), protein, and salt], as well as its protein profile by PAGE analysis, were determined. Modifications in particle size distribution upon UHPH were monitored by laser diffraction in the presence and absence of sodium citrate to dissociate the casein (CN) micelles. Microstructural changes in pressure-treated and non-pressure-treated BM and RO BM particles were monitored by confocal laser scanning microscopy. Particle size analysis showed that UHPH treatment significantly decreased the size of the milk fat globule membrane fragments in BM and RO BM. Also, pressure treatment at 300 MPa led to a significant increase in the recovery of total lipids, CN, calcium, and phosphate in the BM soluble fraction (top layer) following ultracentrifugation. However, PL were primarily concentrated in the pellet cloud (middle layer), located above the pellet in BM concentrated by RO. In contrast, PL were evenly distributed between soluble and colloidal phases of BM. This study provides insight into the modifications of sweet BM constituents induced by RO and UHPH from a compositional and structural perspective.

On the use on hydroxyapatite suspensions for the separation of milk fat globule membrane components from buttermilk.

The milk fat globule membrane (MFGM), which surrounds and stabilizes the fat globules, is released in buttermilk during cream churning. MFGM has many health benefits due to its composition rich in phospholipids and membrane proteins. Many techniques have been tried to separate the MFGM from the remaining milk solids non-fat, but they are challenging to carry out at an industrial scale. This research proposes a new approach to separating MFGM from buttermilk. This paper assessed the efficacy of hydroxyapatite (HA) cristal in interacting with MFGM isolates obtained from either raw or pasteurized cream. Different HA to MFGM ratios were used (10:1 and 20:1) to determine the impact of HA concentration on the adsorption. The results showed a very high affinity of the MFGM for HA and suggested the potential for its separation from buttermilk to improve its valorization.

Effect of alkalinization and ultra-high-pressure homogenization on casein micelles in raw and pasteurized skim milk.

Mechanical and physicochemical treatments of milk induce structural modifications of the casein (CN) micelles, affecting their techno-functional properties in dairy processing. Here, we studied the effect of alkalinization and ultra-high-pressure homogenization (UHPH) on CN micelles in raw skim milk (rSM) and pasteurized skim milk (pSM). The pH of both skim milks (approximately 6.7) was adjusted to 8.5 and 10.5 before UHPH at 100, 200, and 300 MPa. The structural changes of the CN micelles during the treatments were assessed using laser diffraction, transmission electron microscopy, and turbidity measurements. Finally, ultracentrifugation (70,000 × g for 1 h at 20°C) was carried out to evaluate the protein's distribution between the supernatant (serum phase) and the pellet (colloidal phase) by gel electrophoresis and protein concentration measurement. Alkalinization of both skim milks induced a significant reduction in turbidity, whereas an increase of the average particle size was observed, the effect being more severe in pSM than rSM. At alkaline pH, more proteins were recovered in the serum phase, which suggested that the CN underwent major rearrangements into nonsedimentable CN forms of various sizes, as confirmed by transmission electron microscopy. The amount of CN found in the serum phase at pH 8.5 also increased with the UHPH pressure. Although UHPH did not influence the average CN micelle size at pH 6.7 and 8.5, a pressure-dependent decrease was observed at pH 10.5 for both skim milks. The structural changes of the CN micelles observed in this study throughout the combination of alkalinization and UHPH could be of interest for developing new dairy ingredients with improved functionality.

Use of X-ray micro tomography to study the microstructure of loose-packed and compacted milk powders.

The use of X-rays to evaluate food microstructure has developed considerably in recent years. In this work we used X-ray micro tomography (XMT) to visualize the microstructure of loose-packed and compacted samples of spray-dried skim milk powder (SMP) and whole milk powder (WMP) and to quantify the proportion of both interstitial and occluded air voids in each sample. Visual scrutiny of the acquired XMT images showed details of the microstructure of the milk powders such as the spherical morphology of the particles, the size of the particles and internal air voids of various sizes. Within loose-packed powders, the proportion of air voids was higher (13% average) in WMP than in SMP. This was reversed in the compacted powders, in which the proportion of air voids was higher in SMP. The disparity in the proportion of air voids in both loose-packed and compacted samples of SMP and WMP was attributed to the size, shape and surface properties of the particles.

Effect of iron saturation on the recovery of lactoferrin in rennet whey coming from heat-treated skim milk.

This study aimed to determine the effect of thermal treatments on the recovery of lactoferrin in whey coming from rennet-coagulated skim milk. The impact of lactoferrin iron saturation was also assessed using skim milk spiked with different lactoferrin iron forms. The recovery of lactoferrin in the rennet whey fraction was determined by reverse-phase HPLC. One- and 2-dimensional sodium dodecyl sulfate PAGE analyses were performed on rennet curds to characterize the protein interactions involving lactoferrin in heated milk. The extent of lactoferrin recovered in the whey fraction was found to reduce as the heating temperature increased. The binding of iron by lactoferrin improved its thermal stability and its recovery in the whey fraction. Poly-acrylamide gel electrophoresis results showed that the association of lactoferrin in the unheated milk rennet curd involved noncovalent interactions, whereas upon heating, lactoferrin also interacted via an intermolecular disulfide link. Depending on the severity of the heat treatment, lactoferrin aggregates with Cys-containing proteins (beta-lactoglobulin, alpha-lactalbumin, alpha(s2)-casein, and kappa-casein) occurred by intermolecular thiol/disulfide exchange reactions. These noncovalent and covalent interactions explained the lower recovery of lactoferrin in heated milk.

Gender differences in temperature and vascular characteristics during exercise recovery.

Temperature and vascular responses during exercise recovery were examined in men and women of similar age and fitness status (VO2max: 76 +/- 5 vs 73 +/- 5 mL O2 / kg Fat Free Mass x min). Forearm blood flow (venous occlusion plethysmography; FBF), rectal (Trectal) and forearm skin (Tskin) temperatures (degree C) were measured before and every 15 min up to 105 min (t105) during recovery from a 45-min run at 75% of VO2max. Results indicate Trectal decreased to pre-exercise levels within 25 min in men but reached and remained at values lower than baseline between 60 and 105 min of recovery in women. From 90 to 105 min of recovery, Tskin was lower in women than men (t105 : 29.0 +/- 1.3 vs 30.7 +/- 1.5; p <.05). Recovery FBF (mL/100mL x min) was higher in men than women from the start (6.2 +/- 1.9 vs 4.9 +/- 1.9) to the end of recovery (t105 = 1.7 +/- 0.6 vs 2.6 +/- 1.1) (p <.05). Heat flux calculated at the forearm was higher in women and increased throughout the last hour of recovery (p <.05). Further investigations are needed to examine mechanisms underlying failure of post-exercise core and skin temperatures in women to stabilize at pre-exercise levels.

Effects of buffers on milk fatty acids and mammary arteriovenous differences in dairy cows fed Ca salts of fatty acids.

Ten Holstein cows in early lactation were used in a replicated 5 x 5 Latin square design to study the effects of MgO and three buffers added to diets containing Ca salts of canola oil fatty acids. Treatments were 1) control (basal diet; no buffer). 2) 1.1% NaHCO3 plus 1.1% KHCO3, 3) 1.9% NaHCO3, 4) 0.5% MgO, and 5) 2.0% Na sesquicarbonate (percentage of dry matter). The control diet contained 53% grass silage, 43% concentrate, and 4% Ca salts. Body weight, intake, milk yield, and percentages of milk fat, protein, and lactose were unaffected by treatments. Buffers and MgO tended to increase triacylglycerol extraction by the mammary gland and changed the proportions of some fatty acids in milk. Arterial concentrations of acetate and triacylglycerol were correlated with their respective arteriovenous differences. Extraction by the mammary gland was high for acetate (approximately equal to 58.2%), triacylglycerol (approximately equal to 47.3%) propionate (approximately equal to 34.6%), and glucose (approximately equal to 24.3%). Extraction of free fatty acids, phospholipids, or cholesterol was negligible. Mammary triacylglycerol arteriovenous difference tended to be higher than when MgO was fed than when NaHCO3 was fed. Sodium sesquicarbonate, NaHCO3, and the blend of bicarbonate buffers increased C18:2 in milk fat when compared with the control treatment. The concentration of C18:2 in milk fat decreased when MgO was fed, but the ratio of cis-C18:1 to trans-C18:1 increased compared with effects of dietary NaHCO3. Medium-chain fatty acids in milk fat tended to be higher than Na sesquicarbonate than with NaHCO3. Buffers and MgO modified the profiles of fatty acids in milk.

Fatty acid profile and physical properties of milk fat from cows fed calcium salts of fatty acids with varying unsaturation.

Holstein cows (n = 24) averaging 42 d in milk were used in a randomized complete block design during a 4-wk trial. A control total mixed ration (TMR) was compared with TMR supplemented with Ca salts of fatty acids from canola oil, soybean oil, or linseed oil. The three vegetable oils were progressively more unsaturated; the dominant fatty acids were, respectively, cis-delta-9-C18:1, C18:2, and C18:3. Apparent total tract digestibility of dry matter, crude protein, ether extract, and neutral detergent fiber was higher for rations containing Ca salts than for the control ration. Milk yield increased linearly as the unsaturation of the dominant fatty acid in the Ca salts increased. Milk fat percentage was reduced when Ca salts were added to the rations. The addition of Ca salts to the ration decreased the proportions of saturated fatty acids that contained C6 to C16 and increased the proportions of C18:0, cis-delta-C18:1, and trans-delta-11-C18:1 in milk fat. Proportions of C18:2 and C18:3 increased linearly, and cis-delta-9-C18:1 decreased linearly, as the unsaturation of the dominant fatty acid in the Ca salts increased. The proportion of fat that was liquid at 5 degrees C was higher for butter from cows fed diets containing Ca salts, but the proportion of liquid fat at 20 degrees C was not affected. Calcium salts of unsaturated fatty acids added to the diets of dairy cows improved the thermal properties of milk fat.

Timothy grass or alfalfa silage for cows in midlactation: effect of supplementary barley.

Twenty-four multiparous Holstein cows averaging 104 d of lactation were used in a trial with a split-plot design to evaluate the nutritive value of two silages, timothy grass or alfalfa, both treated with formic acid and stored in plastic bag silos. Silages were offered for ad libitum intake either alone or with 17 or 34% (dry matter basis) dry-rolled barley. Both silages contained similar amounts of acid detergent fiber (ADF) (27.5 and 26.7% for timothy grass and alfalfa, respectively). After 110 d of storage, alfalfa silage contained higher amounts of organic acids and NH3 N but had lower soluble N. Total dry matter intake (DMI) and silage DMI were similar between cows fed both silages. Increased barley proportion decreased silage DMI (19.2 to 14.2 kg/d). Apparent total tract digestibility of dry matter was unaffected by treatment. The digestibility of neutral detergent fiber and ADF was higher for the timothy grass silage than for alfalfa silage and was unaffected by the barley percentage added to either silage. Milk yield was lower (23.9 to 22.6 kg/d) for cows fed the highest proportion of barley. The 4% fat-corrected milk yield was unaffected by treatment. Percentages of fat, protein, and total solids in milk were higher for cows fed diets with the higher barley content. Milk fat and protein yields were similar among treatments. Urea in blood was lower for cows fed timothy grass silage than for cows fed alfalfa silage (4.68 vs. 6.23 mg/100 ml). These results suggest that timothy grass silage and alfalfa silage, when stored at a similar ADF content, have comparable nutritive value for midlactation cows.

Dietary soybeans extruded at different temperatures: milk composition and in situ fatty acid reactions.

Twenty-four multiparous Holstein cows, 4 of which were ruminally fistulated, were assigned to one of four diets containing full-fat soybeans, either raw or extruded at 120, 130, or 140 degrees C. Our hypothesis was that the extrusion of full-fat soybeans, as well as the extrusion temperature, would affect the bypass of fatty acids in the rumen and, thus, would modify the fatty acid profile of milk fat. Total mixed diets containing 23.7% soybeans (percentage of DM) were fed for 8 wk. Milk yield was lower, and the proportion of milk CP was higher, for cows fed raw soybeans than for cows fed extruded soybeans. Compared with raw soybeans, extruded soybeans increased the concentration of delta-11-trans-C18:1 from 2.72 to 11.41% in milk fat but had no effect on yield or percentage of milk fat. Polyunsaturated fatty acids of raw soybeans disappeared more rapidly than did those of extruded soybeans from bags incubated in the rumen of fistulated cows. However, more delta-11-trans-C18:1 and C18:0 appeared in bags containing extruded soybeans than in bags containing raw soybeans. Extrusion of full-fat soybeans influenced the metabolism of fatty acids in the rumen and the fatty acid profile of milk fat, but the temperature of extrusion had only minor effects on these parameters.

Lactational response of cows to different concentrations of calcium salts of canola oil fatty acids with or without bicarbonates.

Holstein cows (n = 24) averaging 39 d of lactation were used in a randomized complete block design during an 8-wk trial. From wk 1 to 4, diets contained 62% alfalfa silage and 38% concentrates (dry matter basis), and, from wk 5 to 8, diets contained 47% forage and 53% concentrates. The concentrates were increased for the second phase so that the effect of bicarbonates could be expressed more fully. Diets 1, 2, and 3 contained 2% of a blend of Na and K bicarbonates and 0, 2, or 4% of Ca salts of canola oil fatty acids (percentage of dry matter), respectively. Diet 4 contained the same percentage of Ca salts as did diet 3 but without bicarbonates. Dry matter intake decreased linearly (wk 4), and milk yield was altered quadratically (wk 4), as the percentage of Ca salts in the diet increased. Milk fat percentage (wk 8) and yield (wk 4 and 8), as well as milk protein percentage (wk 4 and 8) and yield (wk 4), decreased linearly as the percentage of Ca salts in the diet increased. Short- and medium-chain saturated fatty acids decreased linearly, and C18:0, trans-delta-11-C18:1, cis-delta-9-C18:1, cis-delta-11-C18:1, and C18:2 increased linearly, as Ca salts in the diet increased. Addition of Na and K bicarbonates to the diets that contained Ca salts increased milk and milk protein yields and increased the proportions of C18:2 in milk fat at wk 8. Dietary bicarbonates had no effect on the responses of other milk fatty acids to supplementation of 4% Ca salts of canola oil fatty acids.

Oxidation of 13C-glucose and 13C-fructose ingested as a preexercise meal: effect of carbohydrate ingestion during exercise.

The oxidation of 13C-labeled glucose and fructose ingested as a preexercise meal between 180 and 90 min before exercise was measured on 6 subjects when either a placebo or sucrose was ingested during the exercise period. Labeled hexose oxidation, which occurred mainly during the first hour of exercise, was not significantly modified when sucrose was ingested, but exogenous glucose oxidation was significantly higher than exogenous fructose oxidation in both situations. The results suggest that the absorption rate of exogenous hexoses was high when exercise was initiated but diminished thereafter, and that glucose and fructose released from sucrose ingested during exercise did not compete with glucose or fructose ingested before exercise for intestinal absorption, for conversion into glucose in the liver (for fructose), or for uptake and oxidation of glucose in peripheral tissues. However, as already shown, in terms of availability for oxidation of carbohydrates provided by the preexercise meal, glucose should be favored over fructose.

Influence of a transscrotal testosterone propionate administration on the serum level of selected hormones of the hypophyso-gonadal axis.

This study was designed to examine the effect of a percutaneous scrotal administration of testosterone propionate (TP) on selected blood variables, in order to identify a reliable anti-doping probe liable to disclose the illicit use of testosterone. Twelve healthy adult males gave their informed consent for the study. Each morning (8:30) and for 10 consecutive days (D), a placebo (D:1,2,3,...8,9,10) or a testosterone propionate (200mg TP on D:... 4,5,6,7...) scrotal patch was installed. On D2 or D3 (placebo-treated) or D7 (TP-treated), venous blood samples were collected at 5 min intervals from 9:00 until 13:00. Serum LH, FSH, 17alpha-hydroxyprogesterone (17HP), testosterone (T), estradiol (E2) and SHBG contents were analysed by immunoassays. The high sampling frequency revealed that TP was associated with the complete abolition of serum LH pulses. Although statistically significant, TP treatment was not related to explicit changes in serum FSH, E2, T/E2 and T/SHBG. TP-induced effects were most significant on serum LH, T and 17HP and were most clearly illustrated by a bi-dimensional distribution plot of serum values of the latter variables. The expression of a combination of the latter parameters could eventually serve to detect testosterone misusers.

Frequent serum sampling in healthy men discloses testosterone peaks exacerbated by testosterone propionate administration.

Interval samplings uncover blood diurnal oscillations for several hormones, highlighting the importance of short time intervals in the disclosure of subtle pulsatile patterns of some peptide hormones, namely LH. In a study designed to develop new probes against steroid misuse, venous blood was sampled at 5-min intervals for 4 hours from 12 eugonadal adult male athletes, 6 receiving transcutaneous administrations of testosterone propionate and 6 placebo subjects. Brief supraphysiologic serum testosterone peaks were disclosed, the amplitude and frequency of these peaks being larger for the treated group. No solid explanation could be given to explain these bursts. Neither the binding/dissociation kinetics of SHBG molecules with and without increased circulating level of dihydrotestosterone, nor brief testosterone-inducing LH bursts, nor increased Leydig cell release could be invoked to explain these peaks. Their occurrence, although relatively rare, could represent a threat and lead to improper treatment.

Performance and profiles of milk fatty acids of cows fed full fat, heat-treated soybeans using various processing methods.

Twenty-four multiparous Holstein cows (48 d in lactation) were used in an 8-wk trial employing a completely randomized block design to determine the effect of heat treatment of full fat soybeans on the fatty acid composition of milk fat. Treatments consisted of a total mixed ration supplemented with 1) ground raw, 2) extruded, 3) micronized, or 4) roasted soybeans included at 17.5% of the dry matter of the total mixed ration. Dietary treatment had no effect on body weight, yields of milk or solids-corrected milk, fat percentage, or yield of milk fat or protein. Cows fed ground raw soybeans had a higher dry matter intake and a higher percentage of protein in milk than those fed heat-treated soybeans. The concentrations of saturated fatty acids from C8:0 to C16:0 were lower, and cis-delta-9-C18:1 tended to be higher, in the milk fat of cows fed heat-treated soybeans than in the milk fat of cows fed ground raw soybeans. The concentration of trans-delta-11-C18:1 was lowest for cows fed ground raw soybeans, intermediate for cows fed micronized and roasted soybeans, and highest for cows fed extruded soybeans. Cows fed extruded soybeans had lower concentrations of C18:2 and C18:3 than did cows fed micronized or roasted soybeans; cows fed ground raw soybeans had intermediate concentrations of C18:2 and C18:3.

Effect of short-term creatine supplementation on renal responses in men.

There is an increasing utilisation of oral creatine (Cr) supplementation among athletes who hope to enhance their performance but it is not known if this ingestion has any detrimental effect on the kidney. Five healthy men ingested either a placebo or 20 g of creatine monohydrate per day for 5 consecutive days. Blood samples and urine collections were analysed for Cr and creatinine (Crn) determination after each experimental session. Total protein and albumin urine excretion rates were also determined. Oral Cr supplementation had a significant incremental impact on arterial content (3.7 fold) and urine excretion rate (90 fold) of this compound. In contrast, arterial and urine Crn values were not affected by the Cr ingestion. The glomerular filtration rate (Crn clearance) and the total protein and albumin excretion rates remained within the normal range. In conclusion, this investigation showed that short-term oral Cr supplementation does not appear to have any detrimental effect on the renal responses of healthy men.

Juice-extracted grass pellets and sodium bicarbonate for cows fed timothy silage of two chop lengths.

Eighteen multiparous Holstein cows in midlactation were used in a switchback design trial to evaluate the effect of chop length (3 or 30 mm) of timothy grass silage, containing less than 30% ADF and treated with formic acid, on DMI and cow performance. Within chop length, addition of NaHCO3 (2% of DMI) or the replacement of 30% of silage DM with juice-extracted grass pellets was also evaluated. Cows were fed a TMR composed of 90% silage and 10% concentrate. Silage preservation characteristics were not different between chop lengths. The DMI and apparent digestibility were similar among treatments. Yields of 4% FCM (24.9 vs. 22.7 kg/d), fat (1.03 vs. .93 kg/d), and protein (.83 vs .77 kg/d) were higher with the short chopped silage. Milk protein, milk NPN content, and serum urea were higher for cows fed long chopped silage. Yields of milk and milk constituents were not affected by the addition of juice-extracted grass pellets or NaHCO3. Fat percentage and fatty acid composition of milk remained unchanged by treatments. Reduction of particle size, from 30 to 3 mm, of timothy grass, treated with formic acid at harvest and using compaction at ensiling in bag silos, did not affect silage conservation characteristics but did improve milk, fat, and protein yields when cows were fed high silage diets.