Buttermilk: Much more than

a source of milk phospholipids

V. Conway,*† S.F.Gauthier,* and Y. Pouliot*
*STELA Dairy Research Center, Institute of Nutrition and Functional Food (INAF), Université Laval, Québec, Canada, G1V 0A6
†
Research Center on Aging, Health, and Social Services Center, University Institute of Geriatrics of Sherbrooke, Sherbrooke, Canada, J1H 4C4

et al., 2013; Conway et al., 2013, 2014). The purpose of this review is to
Implications present recent advances in sweet buttermilk utilization and examine its

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• R
 esulting from churning of cream, sweet buttermilk is a source of status as a high-value by-product of dairy processing. The health benefits
unique bioactive molecules capable of modulating cell signaling, of sweet buttermilk MFGM components will be discussed in view of re-
lipid transport, metabolism, and immunity. cent examples from in vitro, in vivo, and clinical studies.
• S
 everal in vitro and in vivo results provide support for the claim of
the cholesterol-lowering action of buttermilk components. The Churning Process and its Consequences:
• C
 linical evidence now confirms the cardiovascular health benefits of The Unique Composition of Buttermilk
short-term consumption of whole buttermilk, due most likely to its
Buttermilk can be obtained through acidification of cream (i.e., cultured
phospholipid content.
buttermilk) or by the churning of cream into butter (i.e., sweet buttermilk).
• B
 uttermilk consumption represents a natural way to manage blood
Cultured buttermilk, the commercially available form of buttermilk, will
pressure and blood lipids among healthy subjects.
not be discussed in this review. Sweet buttermilk, referred simply as but-
• P
 eptides and polar lipids originating from the milk fat globule termilk in the following discussion, is the aqueous fraction resulting when
membrane are most likely responsible for the improved blood cream is churned to make butter. Churning causes the separation of cream
pressure and blood chemistry resulting from buttermilk consumption. (an oil-in water emulsion) into two distinct phases, an aqueous phase called
buttermilk, and an oily phase or dairy fat concentrate (i.e., butterfat). This
Key words: buttermilk, cholesterol-lowering activity, milk fat globule separation happens as a result of mechanical destabilization of the initial
membrane, minor lipids, sphingomyelin emulsion. Contact with air and repetitive physical collisions disrupt the
thin membrane that surrounds and stabilizes the triglyceride globules,
causing globule coalescence (i.e., aggregation) and ultimately, formation
Introduction of a solid phase (i.e., butter) from which buttermilk is separated simply
The minor components of the milk fat globule membrane (MFGM) by draining. Buttermilk is very similar to skim milk in many ways. For
have been associated with various health benefits. As concluded in previ- example, more than 80% of its proteins are major milk proteins, namely
ous reviews (Dewettinck et al., 2008; El-Loly, 2011; Vanderghem et al., caseins and whey proteins. However, its fat composition differs substan-
2011; Contarini and Povolo, 2013), there is much evidence in support of tially from that of skim milk, in amount and composition, due to the pres-
cholesterol-lowering, anti-inflammatory, chemotherapeutic and anti-neuro- ence of MFGM-derived substances. Close to 20% of buttermilk proteins
degenerative effects of MFGM lipids, mainly through the action of the polar are of MFGM origin. These proteins are solubilized during churning, as
lipid portion (i.e., phospholipids). Not only lipids, but also minor proteins illustrated schematically in Figure 1. In fact, any treatment causing disrup-
associated with the MFGM are thought to be important bioactive compo- tion or breakdown of the MFGM will affect the distribution of the minor
nents. Sweet buttermilk, the co-product of butter making, is particularly components of milk-origin in the final matrix. For example, polar lipids
rich in MFGM components. Processes of isolating and purifying buttermilk account for about 0.9% of the total fat content of cream; they make up
MFGM components have been examined in several studies over the past more than 4.5% of buttermilk fat, but only 0.2% of butterfat (Contarini and
decade (Astaire et al., 2003; Morin et al., 2007; Morin et al., 2006; Rombaut Povolo, 2013). The MFGM residues in buttermilk are responsible for the
et al., 2006). The number of scientific publications on the subject of butter- unique nutritional and technological properties of this dairy co-product.
milk components has quadrupled over the past 20 years (PubMed). Table 1 shows the ratio of polar lipids to total fat in various dairy products.
So far, the vast majority of these studies have focused on either frac- The MFGM is a thin structure (10 to 50 nm) but also a complex biophys-
tionating or concentrating various MFGM components, primarily minor ical system that represents 2 to 6% of the milk fat globule total mass (Lopez,
lipids. Meanwhile, the biological effects of the whole sweet buttermilk 2011). In whole milk, fat globules are dispersed throughout the continuous
matrix remain poorly understood. A few well-controlled clinical trials pro- serum phase due to the emulsifying capacity of the MFGM components,
vide support for the buttermilk health-benefits hypothesis (Baumgartner and coalescence is prevented through electrostatic and steric repulsions (Lo-
pez, 2011). To fulfill the function of ensuring the physicochemical stability
© Conway, Gauthier, and Pouliot. of milk, the MFGM is composed of a highly complex mixture of proteins
doi:10.2527/af.2014-0014 and polar and non-polar lipids, which represent more than 90% of its dry

44 Animal Frontiers
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Figure 1. Schematic representation of the different processing steps used in the manufacturing of butter, with their impact on milk matrix structure. (I) Pasteurized cream
is churned in order to induce a phase inversion; (II) the milk fat globule membrane (MFGM) is ruptured; (III) the expelled liquid (i.e., sweet buttermilk) resulting from
churning is drained, and butter grains are further processed.

mass (Spitsberg, 2005). These constituents are arranged in a heterogeneous portant bioactive molecules are delivered to the newborn at the same time
tri-layer structure secreted by the epithelial cells of the mammary glands to promote healthy growth. It is not surprising that MFGM acts as a vehicle
of cattle. The MFGM thus consists principally of 1) a single inner layer of for the transport of important signaling lipids and proteins to the gastro-
polar lipids, 2) a dense protein coat, and 3) an outer bilayer of polar lipids intestinal tract of newborns, to promote the development of their immune
and embedded specific proteins (Vanderghem et al., 2010). and nervous systems as well as proper development of their intestinal and
Proteins are reported to represent 25 to 70% of the MFGM total mass, metabolic functions (Lopez, 2011). The biological activities associated
but those minor MFGM proteins account only for 1 to 4% of all milk with the major MFGM proteins have been reviewed recently (Dewettinck
proteins (El-Loly, 2011). As noted by authors of other reviews, this high et al., 2008) and will not be discussed further in the present paper.
variability is due primarily to the method used for membrane isolation and The MFGM lipids are a complex mixture containing around 70% neu-
analysis. More than 130 specific proteins have now been identified us- tral lipids (primarily triglycerides, di-glycerides, mono-glycerides, choles-
ing contemporary proteomic approaches (Affolter et al., 2010). However, terol esters, and free cholesterol) and 26 to 30% polar lipids, as presented
resolution by sodium dodecyl sulfate polyacrylamide gel electrophoresis in Table 2. The MFGM contains about 60 to 70% of all milk polar lipids
(SDS-PAGE) reveals a small number of major bands. These correspond (Vanderghem et al., 2010). These are largely responsible for the stabil-
to the major MFGM proteins, namely butyrophilin (BTN), xanthine oxi- ity of fat globules in the milk oil/water emulsion, due to their amphiphi-
dase/dehydrogenase (XO/XDH), mucin-like glycoproteins MUC1 and lic nature. Milk polar lipids are divided into two major classes, namely
MUC15, cluster of differentiation 36 (CD36), lactadherin (PAS6/7), adi-
pophilin (ADPH), and fatty-acid-binding protein (FABP). Buttermilk is
particularly rich in BTN, XO/XDH, PAS6/7, ADPH, and FABP (Affolter Table 1. Phospholipid contents of different dairy
et al., 2010). BTN is by far the most abundant, making up about 40% products. Adapted from MacGibbon and Taylor, 2006.
of MFGM protein (Lopez, 2011). Figure 2A provides an example of the Dairy product
major bands resolved by SDS-PAGE, while Figure 2B illustrates the het- Whole Skim
Composition (%, wt/wt) milk milk Cream Buttermilk
erogeneous distribution of proteins in the MFGM tri-layer structure.
Due to their low abundance in the whole milk matrix, MFGM proteins Lipids (a) 4 0.06 40 0.6
are interesting from a bio-functional perspective rather than for their basic Phospholipids (b) 0.035 0.015 0.21 0.13
nutritional value. Although the primary function of milk is nutritional, im- Ratio [(b/a) X 100] 0.9 25 0.5 22

Apr. 2014, Vol. 4, No. 2 45

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Figure 2. (A) Separation of buttermilk proteins (six samples) by sodium dodecyl sulfate polyacrylamide gel electrophoresis (gel concentration 13.5%, STD = molecular
weight standard; Conway et al., 2010). The identified proteins are (top to bottom): mucin-like glycoprotein 1 (MUC1), xanthine oxidase/dehydrogenase (XO/XDH),
cluster of differentiation 36 (CD36), butyrophilin (BTN), lactadherin (PAS6/7), caseins (CN), lactoglobulin (LB), and lactalbumin (LA). (B) Schematic representation of
the multilayer heterogeneous structure of the milk fat globule membrane, according to Lopez (2010, 2011)

glycerophospholipids and sphingolipids. Glycerophospholipids are com- sphingolipids are composed of a backbone (i.e., sphingosine) to which a
posed of a polar head (e.g., ethanolamine, choline, serine, or inositol at- long saturated fatty acid chain is attached to form a molecule called a ce-
tached to a phosphate group) and a glycerol backbone to which C14–C24 ramide. Different units are added to ceramides, namely a sugar unit to form
fatty acid chains are esterified to form the hydrophobic tail of the molecule a cerebroside, an oligosaccharide residue to form a ganglioside, or a phos-
(Figure 3A). Milk fat is typically composed of short-to-medium-length phate group to form sphingomyelin, as shown in Figure 3B (MacGibbon
fatty acid (C4–C14), mostly saturated. However, these fatty acids (C4– and Taylor, 2006). It is interesting that about 97% of all MFGM sphingo-
C14) are fairly absent in milk glycerophospholipids, which are primarily myelins are composed of medium-chain or long-chain saturated fatty acids
composed of medium-to-long unsaturated fatty acids (C18:1, C18:2, and (i.e., C16:0, C22:0, C23:0, and C24:0) with high melting temperatures.
C18:3) (Dewettinck et al., 2008). MFGM matrix fluidity is due to these More rarely, 17% or more of milk sphingomyelins contain C23:0 fatty ac-
longer unsaturated fatty acids (Lopez, 2010). Like glycerophospholipids, ids (Dewettinck et al., 2008). This uncommon characteristic of milk sphin-
gomyelins suggests a specific function, more likely biological, well beyond
its simple structural role in the MFGM. Sphingomyelin is the only phos-
Table 2. Average lipid composition of bovine MFGM.
phorus-containing sphingolipid in milk. Thus, phosphatidylethanolamine,
Based on Keenan and Mather, 2006; Lopez, 2010; phosphatidylcholine, and sphingomyelin are the major milk phospholipids,
and MacGibbon and Taylor , 2006. representing about 90% of all MFGM polar lipids (Table 2).
Lipid class Total lipids (%) The MFGM sphingomyelins are now known to interact with cholesterol
Neutral to form organized rigid domains called lipid rafts, which float in a disorga-
Triglycerides 62 nized fluid matrix of glycerophospholipids (Lopez, 2010). This ability to
Di-glycerides 9 gather cholesterol into ordered domains depends greatly on the saturation of
Mono-glycerides Trace the fatty acid tails. Lipid rafts have important functions in cell signaling, trans-
Sterols 0.2 to 2.0 duction, and intracellular trafficking (Küllenberg et al., 2012). Modification of
Esters 0.1 to 0.3 MFGM lipid composition may therefore greatly affect the biological activity
Free fatty acids 0.6 to 6.0 of buttermilk. Differences in the size and pattern of sphingomyelin-containing
Polar lipid rafts have been reported (Lopez, 2011). The biological significance of
Phospholipids 26 to 31 milk phospholipids will be discussed in detail in the following section.
- Phosphatidylethanolamine 31.1 to 42.0*
- Phosphatidylcholine 19.2 to 34.5*
- Sphingomyelin 17.9 to 34.5*
Health Benefits Associated with Minor Lipids in Milk
- Phosphatidylinositol 4.7 to 6.2* Health benefits of milk phospholipids have been suggested in associa-
- Phosphatidylserine 2.8 to 8.5* tion with cardiovascular disease, inflammation and cancer since the early
* % of total phospholipids. 1900s (Contarini and Povolo, 2013). A recent review examines more than

46 Animal Frontiers
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Figure 3. Structure of milk phospholipids: (A) Glycerophospholipids and (B) sphingomyelin.

100 papers dealing with the health benefits of dietary phospholipids, primar- in modulation of various metabolic pathways. As noted in recent reviews
ily glycerophospholipids (Küllenberg et al., 2012). However, sphingolipids (Küllenberg et al., 2012; Contarini and Povolo, 2013), in vitro and in vivo
are also recognized as highly bioactive compounds capable of exerting their studies suggest a wide range of biological activities of milk phospholipids,
benefits at low concentrations (El-Loly, 2011). Their metabolites (i.e., ce- including anti-cancer and anti-stress, as well as potential for disease preven-
ramides or sphingosine-1-phosphate) are important lipid messengers con- tion. An exhaustive description of the health benefits attributed to MFGM
trolling cell growth and proliferation, apoptosis, and angiogenesis as well polar lipids will not be provided here. Furthermore, most studies of milk
as immune function (Contarini and Povolo, 2013). First discovered in 1884 minor lipids have focused on specific purified components rather than on
par J.L.W. Thudichum in studies of the brain, sphingolipids are important the whole MFGM fraction, and these will not be described in depth either.
structural lipids found in small amounts in various food products including In the section below, we discuss studies published over the past five years
dairy, meat, eggs, and vegetables (Vesper et al., 1999). However, the nature on the positive actions of bovine MFGM, particularly on neurological and
of the sphingolipid backbone, attached fatty acids, and head groups varies immune functions, the proliferation of cancerous cells, and cardiovascular
considerably depending on the food source. For example, plant sphingo- risk, with emphasis on whole buttermilk fraction.
lipids are composed mainly of cerebrosides, while eggs contain almost no
sphingolipid, and MFGM sphingolipids are represented almost entirely by
From In Vitro Data to Clinical Studies
sphingomyelin (Vesper et al., 1999; Lopez, 2010). Of all MFGM constitu-
ents, sphingomyelin is by far the most interesting bioactive component and Sphingomyelin plays an important structural role in all cellular mem-
the most studied. branes, but particularly in the brain cells. In view of their functions in
As mentioned, sphingomyelin is unusually rich in very long saturated brain myelination, dietary phospholipids have been studied as effective
fatty acids compared with glycerophospholipids, in which fatty acids over carriers of essential fatty acids to promote brain health (Küllenberg et al.,
20 carbons long are almost absent. It is this characteristic that allows its 2012). For example, a commercially available fraction of bovine MFGM
close interaction with cholesterol. The lipid rafts thus formed may provide phospholipids (sphingomyelin 8.4%, phosphatidylethanolamine 8.3%,
more fragile points in the MFGM structure (Figure 4). These weakened and phosphatidylcholine 1.9%) has been shown to reduce endoplasmic-
domains may contribute to the biological functions of MFGM during di- reticulum-stress-induced cell death in vitro (Nagai, 2012). Endoplasmic
gestion (Lopez, 2010), possibly by providing binding sites for digestive reticulum stress has been linked to many neurodegenerative disorders,
enzymes and facilitating lipid transport. They might also provide docking including Alzheimer’s disease. The protective action was attributed to
sites on microorganisms and abnormal cells, thus promoting phagocytosis modulation of cell signaling, measured as protein kinase C activation, as
and apoptosis, and facilitate the delivery of important fatty acids (e.g., ome- well as stimulation of autophagocytosis. Moreover, bovine MFGM sphin-
ga-3 fatty acids) to cell-signaling pathways. In newborns, absorbed phos- gomyelin has been shown to promote neurobehavioral development in
pholipids may be incorporated into cell membranes, all over the body, thus premature babies in an eight-week clinical trial (n = 24) using either milk
modifying their composition and the structure of their lipid rafts, resulting sphingolipid or a control treatment with egg sphingomyelin (Tanaka et al.,

Apr. 2014, Vol. 4, No. 2 47

 constitute major binding sites for rotaviruses and other
microorganisms, and any factor that influences milk
fat globule composition (e.g., processing, cow diet,
breed, and stage of lactation) might also affect the
composition, size, and number of these rafts (Lopez,
2011) and thus, their bioactivities. Clinical evidence
supporting a positive action of MFGM-enriched milk
on immunity in healthy children (n = 182) has been
published by Veereman-Wauters et al. (2012). Using a
validated questionnaire, the authors found a significant
reduction in short ( < 3 days) febrile episodes and days
of fever, in association with 4-month consumption of
MFGM-enriched milk. Using the Achenbach System
of Empirically Based Assessment (i.e., standardized

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questionnaires evaluating anxiety, depression, somatic
complaints, social problems, cognitive problems, at-
tention difficulties, rule-breaking behavior, and ag-
gressive behavior), positive changes in behavior
among the treatment group were also noted. Although
no precise mechanisms were proposed, the authors
concluded that dietary MFGM supplementation may
provide clinically significant improvement of both im-
Figure 4. Representation of sphingomyelin (SM)-cholesterol lipid rafts in the milk fat globule
mune and central nervous system functions.
membrane and their biological significance. Adapted from Lopez (2010).
The cholesterol-lowering action of milk phospho-
lipids is one of the most studied health benefits related to
2013). In this study, serum levels of sphingomyelin, and all measured neu-
buttermilk minor components consumption. A paper published in 1979
rodevelopment parameters, were positively associated with the MFGM
(Howard and Marks, 1979) was the first to report a lower concentration
sphingomyelin treatment.
of cholesterol in plasma of subjects consuming cream compared with
The products of sphingomyelin digestion are also known to affect cell
those consuming butter. This fascinating observation led to further in vivo
proliferation, apoptosis, and senescence, based mostly on in vitro studies.
research, which linked the effect first observed by Howard and Marks
Native MFGM isolates from raw milk have been shown to inhibit the pro-
to milk sphingolipids. Indeed, as shown schematically in Figure 5A, the
liferation of HT-29 colon cancer cells in vitro (Zanabria et al., 2013). In
absorption of cholesterol depends almost entirely on its incorporation
view of these effects, the anti-cancer potential of MFGM concentrate ob-
into and dissolution in mixed micelles (Ros, 2000). Furthermore, to be
tained from buttermilk ultrafiltration has been investigated in vivo (Snow
absorbed by intestinal enterocytes, cholesterol must be free for desorp-
et al., 2010). In this study, rats (n = 16 or 17 per group) were fed diets
tion from these micelles. The solubility of cholesterol in mixed micelles
containing either 0.03% (wt/wt) sphingomyelin in corn oil, 0.03% (wt/wt)
depends greatly on the concentration and nature of the phospholipids pres-
sphingomyelin in anhydrous milk fat, or MFGM from buttermilk (0.11%
ent, as well as on the availability of bile salts (Ros, 2000). This depen-
sphingomyelin, wt/wt). Using the aberrant crypt foci (ACF) model, the
dence on micellar solubility explains the cholesterol-lowering action of
buttermilk product was found to protect against colon cancer. The au-
sphingolipids. For example, through interaction with long saturated fatty
thors suggested, based on evidence in the literature, that sphingomyelin
acid chains, cholesterol forms molecular complexes with sphingomyelin
was specifically responsible for the reduction in the number of apopto-
in the intestinal lumen, resulting in a mutual inhibition of their absorp-
sis-resistant cells, predictive of colon cancer. The mechanism proposed
tion (Figure 5B). Slow and incomplete digestion of sphingomyelin in the
was related to sphingomyelin cell-signaling functions, particularly in cell
proximal segment of the intestine, an important site for lipid absorption,
growth, development, and differentiation. To the best of our knowledge,
enhances formation of complexes and reduces cholesterol availability
no clinical evidence of the anticancer potential of buttermilk consumption
(Eckhardt et al., 2002). The effect of raw-cream buttermilk and pasteur-
is actually available in literature, due likely to the difficulty of carrying out
ized-cream buttermilk concentrates, as obtained by microfiltration, on the
long-term cohort studies on humans.
micellar solubility of cholesterol in vitro has been investigated (Conway
Protective effects of dietary sphingolipids against toxins and microor-
et al., 2010). These authors observed a significant reduction (–57.1%) of
ganisms have been proposed in the literature, the active mechanism most
cholesterol solubility in the presence of buttermilk compared with control.
often suggested being competition for binding sites on intestinal mucosal
Although the bioactive component was not clearly identified in this study,
cells. An in vitro study of the anti-infective action of buttermilk and whey
the formation of sphingomyelin-cholesterol complexes was proposed as
cream MFGM concentrates, obtained by microfiltration, has been pub-
an important mechanism modulating cholesterol solubility. It is interest-
lished recently (Fuller et al., 2013). The buttermilk concentrate was found
ing that the effect was much smaller (-17.0%) with pasteurized-cream but-
to be the more potent inhibitor of infection of monkey kidney cells by
termilk. Processing of cream is known to cause the aggregation of mul-
rotavirus. The authors attributed this anti-infective difference between but-
tiple proteins on the MFGM surface, as well as modification of MFGM
termilk and whey cream to variation of their MFGM lipid composition. As
composition (Lopez, 2011). Processing of raw cream or buttermilk (e.g.,
discussed earlier in this paper, sphingomyelin-cholesterol lipid rafts might

48 Animal Frontiers
pumping, air inclusion, homogenization, and temperature changes) likely
affects the ability of the MFGM surface to interact with enzymes, pro-
teins, lipids, or microorganisms. The authors also observed unexpectedly
a greater decrease in cholesterol solubility when the buttermilk was not
fractionated or concentrated through microfiltration. The effect on solubil-
ity was greater for whole buttermilk, followed by buttermilk microfiltra-
tion retentate, and finally microfiltration permeate. In an in vitro study of
the antioxidant properties of buttermilk, similar losses due to fractionation
were observed (Conway et al., 2012).
Clinical evidences of the cholesterol-lowering properties of buttermilk
consumption have been published recently (Baumgartner et al., 2013;
Conway et al., 2013). Short-term consumption significantly reduced
plasma cholesterol and triglyceride concentrations in a double-blind,
randomized, placebo-controlled crossover study on healthy subjects (n =

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34) with mild-hypercholesterolemia (Conway et al., 2013). Participants
underwent both treatments consecutively in random order, consuming ei-
ther 45 g of buttermilk (sphingomyelin = 0.6% of total fat) or 45 g of a
macronutrient-matched placebo (sphingomyelin < 0.1% of total fat) daily
for 4 weeks. The participant then switched to the other treatment (i.e.,
buttermilk or placebo) for an additional 4-week period. A reduction in
serum LDL-cholesterol (–5.6% compared with placebo) was observed in
subjects with greater LDL-cholesterol at screening. The authors attributed
this improvement in blood lipid profile to the inhibitory action of sphin-
gomyelin on intestinal absorption of cholesterol. The highly significant
reduction in blood triglycerides observed in the buttermilk group (-10.7%
compared with placebo) could be attributed to a reduction of triglyceride
hepatic synthesis, resulting from the increase in polar lipids due to but-
termilk consumption, as proposed in a rodent model by Reis et al. (2013).
Baumgartner et al. (2013) studied the effect of the food matrix on cho-
lesterol absorption in a 12-week placebo-controlled study (n = 97). Sub-
jects were assigned to a control group, a group that consumed one egg per Figure 5. (A) The different steps of lipid digestion in the intestinal lumen (adapted
day, or a group that consumed one egg yolk in 100 mL of buttermilk per from Ros, 2000): (1) Arrival of the initial lipid emulsion (i.e., gastric chyme) in the
day. Among women, daily consumption of one egg resulted in increased duodenum; (2) hydrolysis in the duodenum by pancreatic enzymes; (3) formation
plasma cholesterol and LDL-cholesterol compared with the control group, of mixed micelles (i.e., multi-molecular soluble aggregates) and transport of di-
gested lipids to the intestinal microvilli to be further absorbed as monomers; (4) in
while no difference was seen between the control and egg yolk in butter-
the absence of adequate amounts of bile salt, digested lipids form large vesicles that
milk groups. The authors suggested that buttermilk components, mainly are poorly absorbed (PL = phospholipid, TG = triglyceride, MG = mono-glyceride,
sphingomyelin, interfered with intestinal absorption of cholesterol. These FA = fatty acid, DG = di-glyceride) (B) Complex formed between the OH group of
two studies are, to the best of our knowledge, the only available clinical the cholesterol molecule (i.e., polar head) and the amine group of sphingomyelin
investigations of the impact of whole buttermilk on plasma lipids. (SM) and through hydrophobic interaction. This complex inhibits the absorption
of free cholesterol.
Clinical trials conducted using purified sources of sphingolipids have
been published in recent years. An investigation of the acute (n = 29)
and long-term (n = 20) benefits of sphingolipid-enriched buttermilk on al. (1982) reported no modification of blood lipid levels on consumption
the blood chemistry of healthy humans indicated no significant effect on of cultured buttermilk for 3 weeks in a small group (n = 11) of healthy
fasting and postprandial plasma lipid concentrations (Ohlsson et al., 2009, subjects. As explained previously, unlike buttermilk obtained from the
2010). In a randomized crossover study of the cholesterol-reducing poten- churning of cream (i.e., sweet buttermilk), cultured buttermilk is obtained
tial of sphingomyelin (of unspecified source) in 10 healthy subjects, the through acidification. Thus, cultured buttermilk is not enriched in MFGM
supplement did not affect blood lipid profile or cholesterol absorption and components like sweet buttermilk.
synthesis (Ramprasath et al., 2013). Studies involving less than 30 sub- Even if MFGM polar lipids are usually considered as the active compo-
jects with relatively normal LDL-cholesterol concentrations likely limit nents in many studies, the in vivo production of bioactive peptides by diges-
the possibility of observing significant effects. Furthermore, in Ohlsson tion of MFGM material cannot be ruled out as a contributor to the health
et al. (2009, 2010) studies, the beverages used were sterilized at 143°C, benefits observed on buttermilk consumption. For example, it was found
which very likely affected the composition of the MFGM components. through secondary analysis of data published originally by Conway et al.
Although the exact nature of the compound responsible was not iden- (2013) that consumption of buttermilk for 4 weeks significantly reduced
tified in earlier clinical studies on cholesterol-reducing action of whole blood pressure in normotensive subjects compared with control patients
buttermilk fraction (Baumgartner et al., 2013; Conway et al., 2013), its (Conway et al., 2014). Although the exact nature of the blood-pressure-
MFGM origin appears likely. Supporting this assumption, Thompson et reducing principle was not identified, the authors suggested that peptides

Apr. 2014, Vol. 4, No. 2 49

 ponents and matrix surrounding them both seem to
affect their biological activity. For this reason, future
research should view buttermilk as a functional food
rather than as a source of ingredients needing concen-
tration, isolation, or purification by industrial sepa-
ration processes. Since separation methods could not
achieve enrichment of buttermilk polar lipids without
modification of its bioactivity, the production of en-
riched phospholipids milk through modifications of
cow diets must be further investigated as an innova-
tive way to improve buttermilk composition.
From a product development point of view, it could
be of a great interest to conduct clinical investigations
on the synergic action of other well-known cholester-

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ol-reducing or triglyceride-reducing molecules, such
as phytosterols and omega-3 fatty acids, in buttermilk-
based drinks. This could lead to the development of
new and innovative functional foods to prevent car-
diovascular diseases through improvement in blood
chemistry, anti-oxidative stress reduction, and blood
pressure management. Functional food is a growing
and lucrative market. Thus, buttermilk-based bioactive
Sweet buttermilk is the co-product resulting from the churning of cream into butter. This
drinks may represent a solution to the disposal of but-
unique dairy product in enriched in high-value components from the milk fat globule membrane
(MFGM). Courtesy of Valérie Conway. termilk, still seen as a waste effluent of butter making
by dairy industrials.

originating from the digestion of MFGM components could be responsi-

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50 Animal Frontiers
Fuller, K.L., T.B. Kuhlenschmidt, M.S. Kuhlenschmidt, R. Jiménez-Flores, and (INPULSE) enriched formula milk decreases febrile episodes and may improve
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dietary phospholipids. Lipids Health Dis. 11:1–16.
Lopez, C. 2010. Lipid domains in the milk fat globule membrane: Specific role of
sphingomyelin. Lipid Technol. 22:175–178. About the Authors
Lopez, C. 2011. Milk fat globules enveloped by their biological membrane: Unique
colloidal assemblies with a specific composition and structure. Curr. Opin. Col- Dr. Valérie Conway has a Ph.D. in Food
loid Interface Sci. 16:391–404. Science and is currently a Postdoctoral
Fellow in Medicine at the Université de

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MacGibbon, A.K.H., and M.W. Taylor. 2006. Composition and structure of bovine
milk lipids. In: P. F. Fox and P. L. H. McSweeney, editors, Advanced dairy Sherbrooke (Sherbrooke, Canada). Dur-
chemistry: Volume 2. Lipids. Springer, New York. p. 1–43. ing her Ph.D., under the supervision of
Morin, P., R. Jiménez-Flores, and Y. Pouliot. 2007. Effect of processing on the com- Dr. Yves Pouliot, she specialized in the
position and microstructure of buttermilk and its milk fat globule membranes. valorization of buttermilk component,
Int. Dairy J. 17:1179–1187. mainly its lipids fraction, in the prevention
Morin, P., Y. Pouliot, and R. Jiménez-Flores. 2006. A comparative study of the of cardiovascular diseases. From 2007 to
fractionation of regular buttermilk and whey buttermilk by microfiltration. J. 2013, she worked at the Dairy Science and
Food Eng. 77:521–528. Technology Research Centre (STELA) at
Nagai, K. 2012. Bovine milk phospholipid fraction protects Neuro2a cells from Université Laval (Quebec, Canada). She is
endoplasmic reticulum stress via PKC activation and autophagy. J. Biosci. Bio- currently doing research in nutrigenomics,
eng. 114:466–471. under the supervision of Dr. Mélanie Plourde, at the Research Center on
Ohlsson, L., H. Burling, R.D. Duan, and A. Nilsson. 2010. Effects of a sphingolip- Aging. Her interest is prevention of age-related neurological impairments
id-enriched dairy formulation on postprandial lipid concentrations. Eur. J. Clin. through omega-3 fatty acid supplementation.
Nutr. 64:1344–1349.
Ohlsson, L., H. Burling, and Å. Nilsson. 2009. Long term effects on human plasma Dr. Sylvie Gauthier is professor at the de-
lipoproteins of a formulation enriched in butter milk polar lipid. Lipids Health partment of Food Science and Nutrition at
Dis. 8:1–12. Université Laval. She has a strong back-
Ramprasath, V., P. Jones, D. Buckley, L. Woollett, and J. Heubi. 2013. Effect of ground in biochemistry and nutrition. Her
dietary sphingomyelin on absorption and fractional synthetic rate of cholesterol initial expertise was on the comparative
and serum lipid profile in humans. Lipids Health Dis. 12:125. digestion profile of various food proteins
Reis, M.G., N.C. Roy, E.N. Bermingham, L. Ryan, R. Bibiloni, W. Young, L. in model digestion system. Over the years,
Krause, B. Berger, M. North, K. Stelwagen, and M.M. Reis. 2013. Impact of she became recognized for her practical
dietary dairy polar lipids on lipid metabolism of mice fed a high-fat diet. J. expertise in enzymatic hydrolysis of pro-
Agric. Food Chem. 61:2729–2738. teins, production and characterization of
Rombaut, R., J.V. Camp, and K. Dewettinck. 2006. Phospho- and sphingolipid distribu- bioactive peptides from food proteins. Dr
tion during processing of milk, butter and whey. Int. J. Food Sci. Technol. 41:435–443. Gauthier has been involved in the devel-
Ros, E. 2000. Intestinal absorption of triglyceride and cholesterol. Dietary and phar- opment and scale-up of bioactive peptide
macological inhibition to reduce cardiovascular risk. Atherosclerosis 151:357–379. fractions and milk growth factors from dairy products. Dr Gauthier is mem-
Snow, D.R., R. Jimenez-Flores, R.E. Ward, J. Cambell, M.J. Young, I. Nemere, and ber of the STELA Dairy research Center at the Institute of Nutrition and
K.J. Hintze. 2010. Dietary milk fat globule membrane reduces the incidence of Functional Foods (INAF) from Université Laval.
aberrant crypt foci in Fischer-344 rats. J. Agric. Food Chem. 58:2157–2163.
Spitsberg, V.L. 2005. Invited review: Bovine milk fat globule membrane as a poten- Dr. Yvse Pouliot is professor at the de-
tial nutraceutical. J. Dairy Sci. 88:2289–2294. partment of Food Science and Nutrition
Tanaka, K., M. Hosozawa, N. Kudo, N. Yoshikawa, K. Hisata, H. Shoji, K. Shino- at Université Laval. He graduated from
hara, and T. Shimizu. 2013. The pilot study: Sphingomyelin-fortified milk has University Laval in 1987 with a PhD in
a positive association with the neurobehavioural development of very low birth Food Science and Technology. From the
weight infants during infancy, randomized control trial. Brain Dev. 35:45–52. very beginning of his career at the STE-
Thompson, L.U., D.J. Jenkins, M.A. Amer, R. Reichert, A. Jenkins, and J. Kamul- LA Dairy research Center, he focused at
sky. 1982. The effect of fermented and unfermented milks on serum cholesterol. membrane separation processes applied to
Am. J. Clin. Nutr. 36:1106–1111. dairy fluids. The main focus has been the
Vanderghem, C., P. Bodson, S. Danthine, M. Paquot, C. Deroanne, and C. Blecker. production/separation of whey bioactive
2010. Milk fat globule membrane and buttermilks: From composition to valori- peptides, buttermilk minor components
zation. Biotechnol. Agron. Soc. Environ. 14:485–500. in the development of nutraceuticals from
Vanderghem, C., F. Francis, S. Danthine, C. Deroanne, M. Paquot, E. De Pauw, whey and buttermilk. Dr Pouliot has been
and C. Blecker. 2011. Study on the susceptibility of the bovine milk fat globule involved in a large number of university-industry research projects. He is
membrane proteins to enzymatic hydrolysis and organization of some of the currently Director of the Institute of Nutrition and Functional Foods (INAF)
proteins. Int. Dairy J. 21:312–318. from Université Laval.
Veereman-Wauters, G.S. Staelens, R. Rombaut, K. Dewettinck, D. Deboutte, R.- Correspondence: [email protected]
J. Brummer, M. Boone, and P. Le Ruyet. 2012. Milk fat globule membrane

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