Feed restriction affects milk performances and decreases milk lipolysis in dairy ewes.

Spontaneous lipolysis results in the breakdown of milk fat by the lipoprotein lipase (EC: 3.1.1.34), an enzyme present in milk. Free fatty acids (FFAs) and by-products released in milk during lipolysis can alter both the organoleptic value of milk (off-flavors release) and technological properties of dairy products (decrease in creaming capabilities). Current climate change is having significant impacts on the feeding of grazing animals, with negative consequences on the availability and quality of grass. We and others have demonstrated that dietary restriction increases milk lipolysis in the cow species. However, no data about the impact of feed restriction on milk lipolysis is available in the ewe species. Thus, this paper aims to investigate the effect of feed restriction on milk characteristics with regard to lipolysis values in dairy ewes. Two groups of 24 multiparous Lacaune ewes in mid-lactation received a "non-restricted" control diet (100% of ad libitum DM intake) or a "restricted" (RESTR) diet (65% of ad libitum DM intake) according to a 2 × 2 crossover design. Milk gross composition together with lipolysis analyses were performed. Blood samples were also screened for metabolites or hormone concentrations. The RESTR treatment induced a decrease in milk production (- 21% compared with control treatment) and a modification of the metabolism of dairy ewes characterized by an increase in plasma non-esterified fatty acids (NEFAs), which represents the balance between adipose tissue mobilization and the use of NEFA by other tissues (+153%), cholesterol (+17%) and ß-hydroxybutyrate (+4 %) levels. As a result, a decrease in BW of dairy ewes was observed (-7%). Feed restriction also resulted in a decrease in milk lipolysis estimated by the milk FFA measured by the copper-soap method (-63 and -62%, respectively, for morning and evening milking) or by the reference Bureau of Dairy Industry method (-51 and -57%, respectively, for morning and evening milking). The decrease in milk spontaneous lipolysis under feed restriction was not associated with a decrease in lipoprotein lipase activity in ewes. These results will be completed with proteomic and lipidomic studies in milk samples to better understand mechanisms initiated in the ewe species specifically with regard to lipolysis in milk.

Protein signatures of spontaneous lipolysis and lipoprotein lipase activity in cow's milk.

Spontaneous milk lipolysis refers to the breakdown of triacylglycerols in milk. Lipolysis impacts the organoleptic value of milk by causing off-flavours and reduces the technological properties of milk. Lipolysis is caused by lipoprotein lipase (LPL), a tightly regulated enzyme in milk. Our objective was to identify robust biomarkers of lipolysis and putative regulators of LPL enzyme in bovine milk. To achieve this goal, we used feed restriction as a lever to generate highly contrasted samples with regard to milk lipolysis. We combined statistical methods on proteomics data, milk lipolysis and LPL activity values. Following this strategy, we identified CD5L and GP2 as robust biomarkers of high lipolysis in cow milk. We also identified HID1, SURF4 and CUL9 as putative inhibitors of the lipolytic process in the milk. We thus proposed 5 putative biomarkers to be considered in future tools to manage milk lipolysis. SIGNIFICANCE: This manuscript is notable in three aspects. First, this is the first evaluation of the milk proteome relative to milk lipolysis or LPL activity. Second, the relationship between the abundance of proteins and milk traits was evaluated by a combination of univariate and multivariate analyses. Third, we provide a short list of five proteins to be tested in a larger population to feed the pipeline of biomarker discovery.

Co-construction of a method for evaluating the intrinsic quality of bovine milk in relation to its fate.

There are time-tested assessments for the environmental and economic aspects of sustainability. Its societal aspect has mainly been approached through the assessment of animal welfare. However, the intrinsic quality of milk is seldom taken into account. We developed a participatory construction method for the overall assessment of intrinsic milk quality in its different dimensions (sensory, technological, nutritional and health), according to the fate of the raw milk. Two assessment models were developed, for semi-skimmed standardized ultra-high temperature (UHT) milk and for pressed uncooked non-standardized raw milk cheese. They were constructed by a participatory approach involving experts in the dairy sector with the aim to obtain a diagnostic tool that could be used in the field to help farmers to manage the quality of their milk (by prioritizing improvements on major problems). They were shaped from prerequisite specifications (limited costs and time of application, desire to obtain a transparent tool with all the steps kept visible) and current technical and scientific knowledge. They were based on indicators obtained from raw bulk tank milk analyses (30 for UHT milk and 50 for cheese assessments), which were then aggregated into criteria, principles, dimensions and overall intrinsic quality at farm level. The assessment models had parts in common, for example, same four dimensions, common indicators for health and nutritional dimensions. They also had process-specific features: units chosen, criteria, indicators and weightings in relation to the final product specifications. For instance, sensory and technological dimensions are more complex and preponderant in the cheese assessment (three principles for cheese vs one for UHT milk in both dimensions). Another example is the lack of microbial pathogens (as potential health risk for consumer) in the UHT milk assessment because of pasteurization. The assessment models then underwent a sensitivity analysis and an application in 30 farms in indoor and grazing periods to finally obtain overall UHT milk and cheese quality scores at a 1-year level. The tool was found to be applicable at farm level. However, we observed low overall quality scores with a narrow dispersion, characteristic of a severe evaluation. Even so, the assessment models showed up seasonal differences of the UHT milk and cheese quality at both overall and dimensional levels. In the light of new scientific knowledge and future quality objectives, these are adaptable to other dairy products allowing for their specific features.

Evolution of milk composition, milk fat globule size, and free fatty acids during milking of dairy cows.

The objective of this study was to measure milk composition (fat, protein, and calcium contents; fatty acid profile), milk fat globule size, and free fatty acid content throughout milking. Composition was measured from milk samples collected every 1 min during morning milking in 2 previously published experiments. Experiments 1 and 2 used 9 and 6 dairy cows, respectively. From the beginning to end of milking in experiments 1 and 2, lactose content decreased (-0.45 percentage units), as did protein content (-0.28 and -0.17 percentage units, respectively). In contrast, fat content increased (+5.66 and +5.57 percentage units, respectively). Milk fat globule size increased (+1.51 and +0.43 µm, respectively), whereas free fatty acid content (measured after 24 h of storage at 4°C) decreased quickly during the first minutes (-0.45 mEq/100 g of fat from time point 1 to time point 4 in experiment 1, and -0.85 mEq/100 g of fat from time point 1 to time point 5 in experiment 2), and then largely stabilized, with a slight tendency to increase toward the end of milking period in experiment 2 (+0.32 mEq/100 g of fat). The evolution of milk fatty acid composition depended on the experiment. From the beginning to the end of milking, the concentration of C16:0 consistently increased (+3.4 wt/wt % in experiment 1 and +3.3 wt/wt % from time point 2 to time point 7 in experiment 2), whereas the C18:1/C16:0 ratio increased during the first minutes of milking and then slightly decreased (-0.050 in experiment 1 and -0.031 from time point 2 to time point 7 in experiment 2). Calcium content decreased in experiment 2 (-58 mg/kg). In conclusion, milk composition changed greatly during milking, suggesting that different mechanisms are involved in synthesis and excretion, depending on the type of milk component.

Adaptation of dairy cows to increasing degrees of incomplete milk removal during a single milking interval.

Milk accumulation in the udder decreases milk secretion and this effect is explained as well by the effects of the quantity of milk stored in the udder as by the duration and repetition of periods of milk stasis. This experiment aimed to better understand the underlying mechanisms of decreased milk yield in response to the specific effects of the quantity of milk stored in the udder, independent from storage duration, on milk yield and composition. Sixteen Holstein cows were assigned to 4 blocks of 4 cows in a 4 × 4 Latin square design using 7-d periods, with a 4-d sampling period and a 3-d washout period. Cows were milked twice daily at approximately 0700 and 1630 h throughout the trial. Treatments consisted of 4 degrees of milk removal (100, 70, 40, and 0%) applied at one morning milking, designated M0. Effects of the quantity accumulated were studied in relation to udder distension, via measurements of the total distance between the ends of the 4 teats, and cisternal capacity, via the evaluation of cisternal area by ultrasonographic scan at 1 and 9 h after M0. The effect of the quantity accumulated was also evaluated in relation to mammary epithelium permeability by determining plasma lactose concentrations 1 h before and 4, 7, and 10 h after M0. Leaving milk in the udder at M0 decreased milk production during the M0-M1 interval in a negative curvilinear manner. As a result, M0+M1 milk yield decreased or tended to decrease significantly by -1.3, -5.3, and -12.8 kg for the 70, 40, and 0% treatments compared with the 100% treatment (41.7 ± 1.26 kg/d), respectively. Negative carry-over effects on milk yield were observed until the M3 milking only for the 40 and 0% treatments, and no differences were observed between the effects of these treatments. The total distance between teats increased significantly but to decreasing degrees during the M0-M1 interval. For the 40 and 0% treatments, cisternal area, which was increased 1 h after M0 milking, exhibited no further increase during the M0-M1 interval, suggesting cisternal distension was close to maximum. Simultaneously, lactose concentrations increased in blood plasma for only these 2 treatments, and this increase occurred earlier for the 0% treatment. It was also observed that cows presenting the earliest increases in plasma lactose concentrations during milk accumulation lost more milk in response to extended milking intervals.

Characterization of the nongenetic causes of variation in the calcium content of bovine milk on French farms.

Milk is an important source of Ca in Western diets. Milk Ca is important for the cheesemaking process and could be a useful biomarker of Ca regulation in cows. The objective of this study was to identify and quantify nongenetic factors affecting the variation of Ca content in bovine milk. During the PhénoFinLait program, a survey was performed in 3 major areas of milk production in France. This survey consisted of collecting milk samples, together with information about herd management and cow nutrition, from 924 commercial farms. More than 200,000 individual milk samples were collected, and Ca content was measured by mid-infrared spectroscopy. Each farm was surveyed several times during the year, and 3 to 6 milk samples were collected from each cow. An equation to predict milk Ca content from mid-infrared spectra was developed based on the Ca contents of 292 milk samples, and the milk Ca contents of the 200,000 samples were then predicted. Milk Ca content was lowest in Holstein cows, intermediate in Montbéliarde cows, and highest in Normande cows. For all 3 breeds, milk Ca decreased during the first month of lactation and increased after the fourth month of lactation, with the range between minimum and maximum values largest in Holsteins, intermediate in Montbéliardes, and smallest in Normandes. Milk Ca content also decreased with parity in all 3 breeds. By using multiple factorial analysis, 6 major feeding strategies employed on French dairy farms were characterized based on the data from the survey. Calendar month and cow feeding strategy affected milk Ca content, which dropped in the spring during grazing turnout and was lower when cows were fed fresh and conserved grass rather than corn silage. In conclusion, environmental factors induce variations in milk Ca content in addition to the genetics of the cows, which to date have been identified as a main factor of variation of milk Ca content in dairy cows. In several of the tested conditions, increases in milk production and in the amount of Ca daily secreted in milk were associated with a decrease in milk Ca content as though the mammary gland operated to limit the exportation of Ca when milk production rapidly increased. This result would suggest that milk Ca content could be a biomarker of Ca regulation in dairy cows.

Effects of breed, feeding system, and lactation stage on milk fat characteristics and spontaneous lipolysis in dairy cows.

Spontaneous lipolysis (SL) is an enzymatic reaction that leads to a release of fatty acids that can modify technological and sensory properties of milk and milk products. However, few studies have been done to assess the effect of feeding systems (FS) and breed on SL. Most of them were conducted in the 1980s and are not fully representative of cattle today. No previous study investigated the effect of cow breed at the whole-lactation scale. Thus, a trial was carried out to study the effects of 2 FS (high- and low-input FS) with 2 breeds [Holstein (HO) and Normande (NO)] during 1 entire lactation. Sixty-three cows were followed throughout 1 lactation. Cows were divided into 4 groups according to their breed and their FS. The high FS (HFS) consisted of a high-energy diet (in winter, corn silage with 30% concentrate; otherwise, pasture with 4 kg/d of concentrate) and the low FS (LFS) consisted of a low-energy diet (in winter, conserved grass with no concentrate; otherwise, pasture with no concentrate). The cows calved between January and March. Individual milk samples were collected every month from both morning and evening milkings for fat, protein, milk fat globule size, major fatty acids and proteins profiles, and SL determinations. Data were analyzed using the mixed procedure of SAS. The SL was higher in evening milks compared with morning milks. In early lactation, in evening milks, SL was higher in LFS than in HFS. No difference was shown according to the FS in mid and late lactation. Pasture was associated with low SL rate in mid lactation. The NO cows were less susceptible to SL during the entire lactation than HO cows. Finally, early and late lactation periods were identified as being more susceptible to SL, but this depended on breed and FS. During early lactation, HO cows and LFS were associated with higher levels of SL, particularly in evening milks, and, during late lactation, HO cows were associated with higher levels of SL. No intertreatment or intercow correlations (coefficient of determination <0.16) were found between SL, milk fat and protein contents, milk production, milk fat globule size, proportion of fatty acids and proteins, body condition, and weight during the entire lactation. Effects of breed, FS, and lactation stage were clearly identified and quantified. Causal mechanisms might involve energy balance and circadian secretion of milk fat globule components.

Relationships between milks differentiated on native milk fat globule characteristics and fat, protein and calcium compositions.

Many studies have shown that milk fat globule (MFG) diameter varies in dairy cows in relation to diet and/or breed. However, the mechanisms governing the size of the fat globules remain hypothetical. Our objective was to determine the variable biochemical characteristics (fat, protein, fatty acids (FA), casein and calcium (Ca) contents) between individual milk which differed in both MFG diameter and membrane content, in order to speculate about the links between milk synthesis and MFG secretion. With this aim, we built five databases of individual milk samples from 21 experiments performed between 2003 and 2011. Three of them grouped data from trials dealing with breed/diet effects and included information about: (i) MFG size/membrane, fat and protein contents (n=982), (ii) previous parameters plus FA profile (n=529) and (iii) previous parameters plus true protein composition and calcium contents (n=101). A hierarchical clustering analysis performed on these three databases yielded four groups differing in the MFG characteristics. We observed significant differences among groups for the following parameters: (i) fat content and fat : protein ratio; (ii) de novo and polyunsaturated FA contents; (iii) Ca contents. These relationships could result from potential process regulating the synthesis and secretion of MFG: (i) the apical membrane turnover for MFG secretion and (ii) cytoplasmic lipid droplet formation in the lactocyte during its migration from the basal to the apical pole. The two other databases grouped data from trials dealing with milking frequency (n=211), milking kinetics and milk type (residual v. cisternal) (n=224). They were used to study the relationships between the size of the MFG and milk composition for high native fat contents (from 60 up to 100 g/kg in residual milks). We observed curvilinear relationships between the size of the MFG and fat content, as well as with the fat : protein ratio. This result suggests that MFG diameter reaches a threshold but mechanisms are still unknown.

Effects of DGAT1 K232A polymorphism and milking frequency on milk composition and spontaneous lipolysis in dairy cows.

Milk spontaneous lipolysis (SL) of milk triglycerides is induced by the lipoprotein lipase, a milk native enzyme, and leads to an accumulation of partial glycerides and free fatty acids that are responsible for the deterioration of the taste of milk products. The gene coding for diacylglycerol acyltransferase 1 (DGAT1), an enzyme implicated in triglycerides synthesis, has an important polymorphic site at the K232A locus. This gene is well known to modulate milk composition. No data are available on the effects of DGAT1 on SL. Thus, a trial was carried out to evaluate the effects of DGAT1 K232A polymorphism on milk SL upon milking frequency variations [once- (ODM) and twice-daily milking (TDM)]. Twenty-one cows were divided into 3 groups according their DGAT1 K232A genotype: 8 cows had the KK genotype of DGAT1 (KK cows), 8 had the KA genotype (KA cows), and 5 had the AA genotype (AA cows). The trial consisted in 3 successive periods: 3 wk of TDM, 3 of ODM, and 4 of TDM. Samples were collected for fat and protein contents, SL, fatty acid, and protein profiles determinations. The KK cows presented higher fat and protein contents, lower milk production, and higher κ-casein percentage. No significant difference in fatty acid composition was noted between groups. The SL was twice as high for KK cows in TDM situations (1.13 vs. 0.59 and 0.63mEq/100g of fat, respectively, for KK, KA, and AA cows during the first period of TDM, and 0.46 vs. 0.25 and 0.21mEq/100g of fat, respectively, for KK, KA, and AA during the second period of TDM). The SL remained lower in TDM2 than in TDM1. During ODM, no difference in SL was found between groups and SL remained below 0.2mEq/100g of fat. These results demonstrate the existence of a correlation between DGAT1 genotypes and spontaneous lipolysis, in interaction with an environmental factor, milking frequency, although it has not been possible to clarify the causal mechanism at this stage.

Effect of duration of milk accumulation in the udder on milk composition, especially on milk fat globule.

Our objective was to study the effect of duration of milk accumulation on milk fat globules (MFG) secretion to better understand relationships between milk yield, milk fat, and MFG secretion. The modification of the milk accumulation duration in the udder is a tool to increase milk fat content. Four milking frequencies were studied on 6 dairy cows averaging 118±22 d in milk: 2 milkings/d separated by 11- and 13-h or by 4- and 20-h intervals and 1 milking/d. The experimental trial was a double Latin square 3×3 with 2-wk periods. Postexperiment, a milking frequency of 36-h was repeated twice. Compared with 2 milkings with 11- and 13-h frequencies, 1 milking/d reduced milk and milk fat yields and increased fat content, without any effect on the size of MFG. Two milkings with 4- and 20-h intervals had no significant effect on milk fat yield and content but tended to increase the size of the MFG. Lipolysis, measured on morning milk, was weaker with 1 milking/d. When data were analyzed according to milk accumulation duration (4, 11, 13, 20, 24, and 36h), the highest fat content and the largest diameters of MFG were obtained on milks from 4 and 36h milkings (62.8g/kg, 4.15 µm and 57.7g/kg, 4.09 µm, respectively). Such observations could have 2 origins: the richness in residual milk of the 4-h milk and the coalescence of MFG related to the long milk accumulation duration in the 36-h milk. For each duration of milk accumulation, a relationship exists between MFG size and fat yield. The positive relation between MFG size and fat content was confirmed at each duration of milk accumulation. Rate of secretion of milk fat (milk accumulation of 4h excluded) was also well correlated with MFG size. For the 36-h milk, this relationship was also observed but with a significantly different slope, assuming phenomena of MFG coalescence in response to the supposed increased intramammary pressure or to slower secretion rate and, hence, fusion events of microlipids droplets in the cytoplasm. Duration of milk accumulation joined with large increases in milk fat content induces changes in MFG size.

Dietary cation-anion difference and day length have an effect on milk calcium content and bone accretion of dairy cows.

Milk and dairy products are an important source of Ca for humans. Recent studies have shown fluctuations in cow milk Ca content during the year in France, with high values in winter and with corn silage diets, and a decrease during May and June and with grass diets. The aim of this study was to identify the reasons for this seasonal decrease in milk Ca content by testing the effect of 2 levels of dietary cation-anion differences (DCAD; 0 mEq/kg of dry matter for DCAD 0 and 400 mEq/kg for DCAD 400) and 2 day lengths (8 h of light/d for short days: SD; and 16 h/d for long days: LD) on the Ca balances of dairy cows. The DCAD treatments were designed to mimic diets based either on corn silage or on herbage. The cows were only illuminated by solarium lights providing UVA and UVB. The trial was conducted according to 2 simultaneous replicates of a 4×4 Latin square design using 8 dairy cows averaging 103±44 d in milk with 4 periods of 14 d. Data were analyzed by ANOVA with a model including treatment, cow, and period effects. No significant interaction was found between day length and DCAD treatments. With DCAD 400 compared with DCAD 0, blood pH increased and plasma ionized Ca content decreased, whereas the plasma total Ca content did not differ between treatments. Milk Ca content, however, increased with DCAD 400 compared with DCAD 0, in relation to a decrease in the amount of Ca excreted in urine. The DCAD had no significant effect on protein and casein contents and DCAD 400 tended to decrease milk yield. This illustrates that the udder did not decrease Ca uptake from the blood at high DCAD even though DCAD 400 decreased the mammary availability of Ca by decreasing the proportion of blood ionized Ca. Milk Ca and casein contents were significantly lower with LD compared with SD, whereas day length had no effect on milk yield after 14 d of treatment. Bone accretion of cows increased when the Ca content of milk increased (i.e., with DCAD 400 compared with DCAD 0 and with SD compared with LD). This work suggests that long and sunny days could explain part of the seasonal decrease in milk Ca content in summer and refutes the hypothesis that low milk Ca contents at grazing could be due to the high DCAD of herbage.

Primary hematopoietic cells from DBA patients with mutations in RPL11 and RPS19 genes exhibit distinct erythroid phenotype in vitro.

Diamond-Blackfan anemia (DBA) is caused by aberrant ribosomal biogenesis due to ribosomal protein (RP) gene mutations. To develop mechanistic understanding of DBA pathogenesis, we studied CD34⁺ cells from peripheral blood of DBA patients carrying RPL11 and RPS19 ribosomal gene mutations and determined their ability to undergo erythroid differentiation in vitro. RPS19 mutations induced a decrease in proliferation of progenitor cells, but the terminal erythroid differentiation was normal with little or no apoptosis. This phenotype was related to a G0/G1 cell cycle arrest associated with activation of the p53 pathway. In marked contrast, RPL11 mutations led to a dramatic decrease in progenitor cell proliferation and a delayed erythroid differentiation with a marked increase in apoptosis and G0/G1 cell cycle arrest with activation of p53. Infection of cord blood CD34⁺ cells with specific short hairpin (sh) RNAs against RPS19 or RPL11 recapitulated the two distinct phenotypes in concordance with findings from primary cells. In both cases, the phenotype has been reverted by shRNA p53 knockdown. These results show that p53 pathway activation has an important role in pathogenesis of DBA and can be independent of the RPL11 pathway. These findings shed new insights into the pathogenesis of DBA.

Alteration of the nutrient uptake by the udder over an extended milking interval in dairy cows.

Little is known about modifications of the mammary utilization of nutrients circulating in blood plasma when milk yield is strongly decreased by once-daily milking. A trial was carried out to describe the mammary nutritional adjustments linked to the downregulation of milk synthesis as milk accumulated over an extended milking interval in the bovine udder. Three Holstein dairy cows yielding 34.0 kg/d of milk were fitted with an ultrasound flow probe around the left external pudic artery and with catheters inserted into the left carotid and milk vein to estimate mammary blood flow (MBF) and mammary uptake of acetate, ß-hydroxybutyrate, nonesterified fatty acids, glycerol, glucose, O(2), and CO(2) release. The trial was carried out over 2 consecutive weeks, with wk 2 repeating wk 1. Cows were milked twice daily at 12-h milking intervals. On d 3, cows were milked at 0630 h and were not milked for 36 h until d 4 at 1830 h. Over the following days, twice-daily milking was resumed using 12-h milking intervals. Each half-udder was milked separately. Secretion rates of milk and milk proteins decreased 67% during the 12-to-36-h interval of milk accumulation, whereas that of milk fat fell 30%. Timing of changes in MBF and lactose levels in blood plasma was concomitant and significant after 19.5 and 21.5h of milk accumulation in the udder, respectively. The MBF decreased, most likely because the usual increases in MBF no longer occurred when the udder was full of milk. After 24h of milk accumulation, MBF did not increase further when cows lay down, and did not increase as usual 3h after a meal, suggesting a possible physical effect of milk accumulated in the udder on MBF, complementing metabolic regulation. Mammary uptake or release of nutrients was lowered before 24h for glucose, acetate, and ß-hydroxybutyrate and after 24h for total glycerol, O(2), and CO(2), mostly associated with the impaired MBF. However, these decreases ranged from 12 to 17%, and cannot entirely explain the -45 and -20% decreases in milk secretion rates observed during the entire 36 h of milk accumulation, thus confirming the primary role of intramammary metabolic regulation in the downregulation of milk secretion. The larger amount of nutrients taken up by the udder could explain the enhanced milk fat levels, involving a strongly modified metabolic fate of nutrients.

Combined effects of trans-10,cis-12 conjugated linoleic acid, propionate, and acetate on milk fat yield and composition in dairy cows.

Diets inducing milk fat depression (MFD) are known to alter ruminal lipid metabolism, leading to the formation of specific isomers [such as trans-10,cis-12 conjugated linoleic acid (CLA)] that inhibit milk fat synthesis in lactating dairy cows. However, ruminal outflow of these isomers does not fully account for the decreases in milk fat synthesis observed during diet-induced MFD. The high-concentrate diets inducing MFD also induce a greater production of propionate, suggesting a possible inhibition of milk fat by propionate associated with trans-10,cis-12-CLA during MFD. The present experiment aimed to study the combined effects of propionate and trans-10,cis-12-CLA (both inhibitors of milk fat synthesis) on milk fat secretion and the effects of the combination of 2 nutrients with opposite effects (acetate and propionate). Six Holstein cows were used in a 6×6 Latin square design with 21-d periods (14 d of nutrient infusion). The treatments were control; ruminal infusion of 1,500 g/d of acetate (A); ruminal infusion of 800 g/d of propionate (P); duodenal infusion of 1.60 g/d of trans-10,cis-12-CLA (CLA); ruminal infusion of 750 g/d of acetate+400 g/d of propionate (A+P); and duodenal infusion of 1.60 g/d of trans-10,cis-12-CLA+ruminal infusion of 800 g/d of propionate (CLA+P). The amounts of nutrients infused were chosen to induce a similar variation in milk fat content. Treatments A and P decreased dry matter intake. Compared with the control, P and CLA treatments decreased milk fat content and yield by 9% and 15% on average. Treatment A increased milk fat content by 6.5% but did not modify milk fat yield (because of a decrease in milk yield). The effects of A and P, and CLA and P on milk fat and fatty acid percentages and yield were additive (A+P and CLA+P treatments). With a same dose of trans-10,cis-12-CLA, the additional supply of propionate induced a decrease in milk fat 40% higher than that induced by trans-10,cis-12-CLA alone. The milk fatty acid profile obtained with CLA+P was similar to those observed with high-concentrate diets inducing MFD. In conclusion, under our experimental conditions, the effects of the 3 nutrients were additive on mammary lipogenesis, regardless of their separate effects. We also show that propionate could contribute to the milk fat reductions unaccounted for by trans-10,cis-12-CLA during MFD induced by high-concentrate diets.

Linear relationship between increasing amounts of extruded linseed in dairy cow diet and milk fatty acid composition and butter properties.

The aim of this experiment was to compare the effects of increasing amounts of extruded linseed in dairy cow diet on milk fat yield, milk fatty acid (FA) composition, milk fat globule size, and butter properties. Thirty-six Prim'Holstein cows at 104 d in milk were sorted into 3 groups by milk production and milk fat globule size. Three diets were assigned: a total mixed ration (control) consisting of corn silage (70%) and concentrate (30%), or a supplemented ration based on the control ration but where part of the concentrate energy was replaced on a dry matter basis by 2.1% (LIN1) or 4.3% (LIN2) extruded linseed. The increased amounts of extruded linseed linearly decreased milk fat content and milk fat globule size and linearly increased the percentage of milk unsaturated FA, specifically alpha-linolenic acid and trans FA. Extruded linseed had no significant effect on butter color or on the sensory properties of butters, with only butter texture in the mouth improved. The LIN2 treatment induced a net improvement of milk nutritional properties but also created problems with transforming the cream into butter. The butters obtained were highly spreadable and melt-in-the-mouth, with no pronounced deficiency in taste. The LIN1 treatment appeared to offer a good tradeoff of improved milk FA profile and little effect on butter-making while still offering butters with improved functional properties.

Winter feeding systems and dairy cow breed have an impact on milk composition and flavour of two Protected Designation of Origin French cheeses.

This study investigates the effects of two feeding systems and two dairy cow breeds on milk yield and composition, physical and sensorial properties of Camembert and Pont-l'Evêque cheeses. The experiment consisted of a 2 × 2 factorial arrangement of treatments. A low energy grass diet with only 15% of concentrate (LowGS) was compared with a high-energy maize silage diet with 30% concentrate (HighMS). Thirty-four Holstein (Ho) and 34 Normande (No) cows in early lactation were assigned to one of two feeding systems for a 6-week period. Cows on the LowGS feeding system had lower milk yield, fat and protein content. In both feeding systems, No cows had lower milk yields but higher milk protein contents than Ho cows. The LowGS feeding system altered milk fatty acid (FA) composition by reducing saturated FA. Breed had only a small impact on milk FA. Concerning milk coagulating properties, only the firmness was reduced by the LowGS feeding and the Ho breed. The effects of breed and feeding system on the protein content of cheeses were more marked in Camembert cheese than in Pont-l'Evêque cheese. However, the Camembert cheese from Ho-LowGS was, in fact, characterized especially by lower protein content. LowGS feeding system and No breed produced more yellow cheeses. Feeding systems had limited effects on the firmness of Camembert and Pont-l'Evêque cheeses measured by penetrometry. In sensory analysis, Ho breed and LowGS feeding produced a Camembert cheese with a more melting texture in the mouth due to the increase of spreadability index and of proteolysis. The type of cheese also had an influence: the effects were more important on Camembert cheese than on Pont-l'Evêque cheese. Only the Ho-LowGS treatment produced a very specific Camembert cheese different from the others. The feeding systems and breed of dairy cow have no determinant effect on PDO (protected designation of origin) Camembert and Pont-l'Evêque cheeses, especially regarding taste. In this kind of trial, despite the effects of feeding systems and breed on milk composition, the role of cheese ripening and microbiology appears to be of considerable importance.

Effects of feeding camelina (seeds or meal) on milk fatty acid composition and butter spreadability.

The nutritional and rheological properties of butter depend on the fatty acid composition of milk. Therefore, feeding oilseeds rich in unsaturated fatty acids is likely to affect butter properties. The aim of this trial was to examine to what extent feeding the linolenic acid-rich cruciferous plant camelina can affect the fatty acid composition of dairy products and the properties of butter. A control diet composed of 60% corn silage-based ration and completed with high-energy and nitrogenous concentrates was compared with 2 experimental diets designed to provide the same amount of polyunsaturated fatty acids via either camelina seed (630 g/d, CS diet) or camelina meal (2 kg/d, CM diet). The diets were isoenergetic and isonitrogenous. The trial followed a double 3 x 3 Latin-square design with 4-wk periods on 6 Holstein dairy cows. The camelina diets tended to decrease dry matter intake but did not have a significant effect on milk production. They generated a slight decrease in milk protein and a strong decrease in milk fat yield and content. The CM diet led to a stronger decrease in fat content. Camelina generated a greater proportion of monounsaturated fatty acids, notably C18:1 trans isomers, including trans-10 and trans-11 C18:1, which increased by 11.0- and 2.6-fold, respectively, with the CM diet. Camelina also led to an increase in conjugated linoleic acids, particularly rumenic acid, cis-9, trans-11 C18:2. Camelina did not affect parameters of buttermaking except churning time with milk from CM fed cows, which was longer. The butters of camelina diets were softer at all temperatures tested, especially with the CM diet. In conclusion, feeding camelina can modify milk fatty acid profile and butter spreadability.

Glutamine stimulates translation of uncoupling protein 2mRNA.

Uncoupling protein 2 (UCP2) belongs to a family of transporters/exchangers of the mitochondrial inner membrane. Using cell lines representing natural sites of UCP2 expression (macrophages, colonocytes, pancreatic beta cells), we show that UCP2 expression is stimulated by glutamine at physiological concentrations. This control is exerted at the translational level. We demonstrate that the upstream open reading frame (ORF1) in the 5' untranslated region (5'UTR) of the UCP2 mRNA is required for this stimulation to take place. Cloning of the 5' UTR of the UCP2 mRNA in front of a GFP cDNA resulted in a reporter gene with which GFP expression could be induced by glutamine. An effect of glutamine on translation of a given mRNA has not been identified before, and this is the first evidence for a link between UCP2 and glutamine, an amino acid oxidized by immune cells or intestinal epithelium and playing a role in the control of insulin secretion.

Composition of milk fat from cows selected for milk fat globule size and offered either fresh pasture or a corn silage-based diet.

The objective of this study was to examine the synthesis and composition of milk produced by dairy cows that secrete either small milk fat globules (SMFG) or large milk fat globules (LMFG), and to study their response to diets known to alter milk composition. Four groups of 3 multiparous dairy cows were assigned to 2 isoenergetic feeding treatments: a corn silage treatment supplemented with soybean meal, and fresh pasture supplemented with cereal concentrate. The 4 groups comprised 2 groups of 3 dairy cows that produced SMFG (3.44 microm) and 2 groups of 3 dairy cows that produced LMFG (4.53 microm). The SMFG dairy cows produced higher yields of milk, protein, and calcium. Nevertheless, their milk had lower fat and protein contents. Both SMFG and LMFG cows secreted similar amounts of milk fat; therefore, higher globule membrane contents in milk fat were observed in SMFG cows. Higher calcium mineralization of the casein micelles in SMFG cows suggests that it may be possible to improve cheese-making properties even if the lower protein content may lead to lower cheese yields. The SMFG cows secrete milk fat with a higher concentration of monounsaturated fatty acids and a lower concentration of short-chain fatty acids. They also have a higher C18:1/C18:0 ratio than LMFG cows. This suggests that SMFG cows have more significant fatty acid elongation and desaturation. The pasture treatment led to an increase in milk and protein yields because of increased energy intake. It also resulted in lower milk fat yield and fat and protein contents. The pasture treatment led to a decrease in milk fat globule size and, as expected, an increase in monounsaturated and polyunsaturated fatty acid contents. However, it induced a decrease in the protein content, and in calcium mineralization of casein micelles, which suggests that this type of milk would be less suitable for making cheese. This study also shows that there is no correlation between the cows, based on milk fat globule size and diet. These results open up possibilities for improving milk fat quality based on milk fat globule size, and composition. The mechanisms involved in milk fat globule secretion are still to be determined.

Translation control of UCP2 synthesis by the upstream open reading frame.

Uncoupling protein 2 (UCP2) belongs to a family of transporters of the mitochondrial inner membrane. In vivo low expression of UCP2 contrasts with a high UCP2 mRNA level, and induction of UCP2 expression occurs without change in mRNA level, demonstrating a translational control. The UCP2 mRNA is characterized by a long 5' untranslated region (5'UTR), in which an upstream open reading frame (uORF) codes for a 36-amino-acid sequence. The 5'UTR and uORF have an inhibitory role in the translation of UCP2. The present study demonstrates that the 3' region of the uORF is a major determinant for this inhibitory role. In this 3' region, a single-base substitution that kept the codon sense unchanged significantly modified UCP2 translation, whereas some important amino acid changes had no effect. We discuss our results within the framework of the existing models explaining initiation of translation downstream of a uORF.