t-s

Grki

relanMatPolaent

4 April 2014Accepted 4 April 2014

as well as high CLA content with signicant amount of CLA9.Moreover, milk fat inuences processability of the rawmaterial andis a carrier of taste and aroma.

trzak-Fiecko et al.,jtowski, Dankw,a, & Bia1ek, 2011).cerning fatty acidery advantageousn. Investigation inncentration of a-atio favouring ALAPikul et al., 2008).erol was lower incan be favourablekul & Wjtowski,

vitamins. However, it transpires from a review of available litera-ture that few research teams have conducted comprehensive anddetailed analysis of vitamins A, D, E, K and b-carotene in maresmilk (Csap et al., 1995; Marconi & Panli, 1998). The aim of thecurrent study was to add to the body of knowledge on mares milk,the principal information about fatty acid prole, fat and choles-terol as well as vitamins A, D, E, K and b-carotene content in latelactation, machine milked, mares milk.

* Corresponding author. Tel.: 353 1 601 2143.E-mail addresses: maria.markiewicz-keszycka@ucd.ie, maria.kesz@gmail.com

Contents lists availab

International D

.e l

International Dairy Journal 38 (2014) 31e36(M. Markiewicz-Keszycka).and applies to high fat soluble vitamins and n3 fatty acid content It is commonly believed that milk contains a lot of fat-solubleAlthough mares milk production does not have such economicimportance and long traditions as production of milk from rumi-nants, it is gaining more importance in European countries. This isdue to the fact that thanks to its composition, mares milk can be aprotable alternative to cows milk especially among people withcows milk allergy, the elderly and convalescent (Salimei & Fantuz,2012; Uniacke-Lowe, 2011; Uniacke-Lowe, Huppertz, & Fox, 2010).

The rst part of our current studies presented the characteristicsof the whey protein fraction of late lactation mares milk(Markiewicz-Keszycka et al., 2013a); this manuscript describes thelipid fraction of late lactation mares milk. According to Palupi,Jayanegara, Ploegera, and Kahla (2012), the premium nutritionalquality of dairy products is highly correlated with milk fat quality

& Csap-Kiss, 1995; Marconi & Panli, 1998; Pie2009; Pikul & Wjtowski, 2008; Pikul, WKuczynska, & Lojek, 2008; Rutkowska, AdamskHowever, there are relatively few studies conprole in the last stage of lactation, which is a vperiod for obtaining milk for human consumptiothis area showed that mares milk has high colinolenic acid (ALA) and linoleic acid (LA), at a r(Csap et al., 1995; Pietrzak-Fiecko et al., 2009;Moreover, the concentration of fat and cholestmares milk than in human or cows milk, whichfor people with low-fat diet requirements (Pi2008).1. Introduction In recent years, the fatty acid composition of mares milk hasbeen examined by different authors (Csap, Steer, Martin, Makray,http://dx.doi.org/10.1016/j.idairyj.2014.04.0030958-6946/ 2014 Elsevier Ltd. All rights reserved.acids composition, fat soluble vitamins and b-carotene content were measured. The study revealed thatmares milk is a low fat product with low cholesterol content. The lipid fraction of mares milk wascharacterised by a low concentration of saturated fatty acids, as well as low ratio of linoleic and linolenicfatty acids. The results obtained showed that, despite its low fat content, mares milk is a good source offat-soluble vitamins, especially vitamins D3, E and K2.

2014 Elsevier Ltd. All rights reserved.Received 5 January 2014Received in revised formduced by Polish Cold-blooded mares between 141 and 210 d post partum. Total fat, cholesterol, fatty

Article history: The aim of this research was to study the lipid fraction composition of late lactation mares milk pro-Short communication

Concentration of selected fatty acids, fab-carotene in late lactation mares milk

Maria Markiewicz-Keszycka a,*, Jacek Wjtowski b,Beata Kuczynska c, Kamila Puppel c, Jzef Krzy _zewsArtur Jzwik d, Emilia Bagnicka d

a School of Agriculture and Food Science, University College Dublin, Beleld, Dublin 4, Ib Poznan University of Life Sciences, Department of Small Mammals Breeding and RawcWarsaw University of Life Sciences, Department of Animal Breeding, 02-787 Warsaw,d Institute of Genetics and Animal Breeding of the Polish Academy of Sciences, Departm

a r t i c l e i n f o a b s t r a c t

journal homepage: wwwoluble vitamins and

a _zyna Czy _zak-Runowska b,d, Nina Strza1kowska d,

derials of Animal Origin, Zotniki, 62-002 Suchy Las, Polandndof Animal Sciences, Jastrzebiec, 05-552 Magdalenka, Poland

le at ScienceDirect

airy Journal

sevier .com/locate/ idairyj

ate

y ac

201ang

natiTable 1The fat and cholesterol content and fatty acid composition of mares milk from the l

Itema Mean (SD) Minimum

Total fat (g L1) 4.2 (1.8) 2.4Cholesterol (mg L1) 2.04 (0.93) 1.22Fatty acid (g 100 g1 of fat)C8:0 2.54 (0.51) 1.54C10:0 4.52 (0.50) 3.47C10:1 1.17 (0.24) 0.78C12:0 4.25 (0.90) 3.34C14:0 5.43 (0.59) 4.32C14:1 0.50 (0.05) 0.45C16:0 20.53 (1.23) 18.56C16:1 cis-9 5.51 (0.64) 4.08C18:0 2.09 (0.60) 1.32C18:1 cis-9 25.04 (1.34) 21.96C18:1 cis-11 1.37 (0.18) 1.18C18:2 n6 LA 3.81 (1.11) 2.76C18:3n3 ALA 17.51 (3.03) 11.83SFA 49.35 (3.37) 34.35MUFA 33.59 (1.54) 30.55PUFA 21.33 (2.39) 17.41LA/ALA 0.23 (0.11) 0.14PUFA/SFA 0.55 (0.10) 0.39

AI 0.85 (0.13) 0.66TI 0.38 (0.09) 0.29

a Abbreviations are: LA, linoleic acid; ALA, a-linolenic acid; SFA, saturated fattatherogenic index; TI, trombogenic index.

b Strza1kowska et al. (2009).c Moate, Chalupa, Boston, and Lean (2007).d Puppel, Na1ecz-Tarwacka, Kuczynska, Go1ebiewski, and Grodzki (2012b).e Darragh (2002).f Koletzko et al. (2001); Koletzko, Agostoni, Bergmann, Ritzenthaler, and Shamir (g Arsic et al. (2009); Molt-Puigmart et al. (2011); Wan, Wang, Xu, Geng, and Zh

M. Markiewicz-Keszycka et al. / Inter322. Materials and methods

2.1. Animals, feeding and milk sample collection

The experiments were carried out on individual milk samplesfrom 9 Polish Cold-blooded (CBHs) multiparous mares reared onan equine dairy farm in the Wielkopolska region (WesternPoland). All animals were in the late stage of lactation between141 and 210 d post partum. The average body weight of mareswas 743 kg. The age of experimental animals ranged from 9 to14 y. The sample collection procedure, feeding and milkingtechniques were described previously (Markiewicz-Keszyckaet al., 2013a). In brief, the mares were machine-milked,without the use of oxytocin, after being separated from theirfoals forw3 h. Representative bulk milk samples from each marewere collected for laboratory analysis. Milk samples for total fat,vitamins and cholesterol analysis were collected twice: once atthe beginning of October and once at the beginning of Novemberof 2011 (n 18 samples), while samples for fatty acids compo-sition were collected only once, in November 2011 (n 9samples).

2.2. Experimental procedures

For examination of the milk fat content, extraction of fat wasperformed according to Rse-Gottlieb procedure (AOAC, 1990) atroom temperature. Fatty acid methylation was performed accord-ing to the transesterication method by EN ISO 5509 (2000). Theconcentration of fatty acids was determined according to themethod of Kuczynska et al. (2012), using a Hewlett Packard (HP)6890 gas chromatograph with HP ChemStation software (HewlettPackard/Agilent, Waldbronn, Germany), a ame ionisation detectorand a Varian Select FAME column (100m length, 0.25mmdiameter,stage of lactation in comparison with cow and human milk.

Maximum Cow (SD) Human

8.4 39.8e44.3b 30.0e50.0e

4.72 155.1e195.8b 100.0e200.0f

3.25 1.17 (0.35)c 0.06e0.21g

4.98 2.480 (0.73)c 0.65e1.39g

1.61 nd nd6.24 2.99 (0.850)c 3.59e6.48g

6.18 10.38 (1.71)c 3.92e7.44g

0.60 1.08 (0.36)c 0.16e0.24g

22.39 28.51 (4.98)c 18.68e22.24g

6.45 1.73 (0.63)c 0.44e2.50g

3.03 10.51 (3.59)c 5.63e6.45g

26.70 20.5 (5.35)c 31.26e37.45g

1.68 nd nd5.58 3.13 (2.11)c 15.24e17.73g

21.88 0.59 (0.36)c 0.6e1.36g

44.90 53.67e63.52d 38.52e44.3g

35.64 30.12e38.19d 36.56e42.92g

24.94 3.63e5.84d 18.24e19.1g

0.47 5.30c 25.40e33.24g

0.71 0.06e0.10d 0.47g

1.08 1.63e2.30d 0.74g

0.56 2.12e2.83d 0.68e1.0g

ids; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids; AI,

1).(2009).

onal Dairy Journal 38 (2014) 31e360.25 mm lm thickness; Varian/Agilent Technologies, Waldbronn,Germany).

Fatty acid separation was performed at a pre-programmedtemperature: 130 C for 1 min; 130e170 C at 6.5 C min1; 170e215 C at 2.75 C min1; 215 C for 12 min, 215e230 C at20 C min1 and 230 C for 3 min. The ow rate of the carrier gaswas equal to 25 cm s1. Other parameters were: carrier gas (He)ow-constant (1.2 mL min1), injector temperature 240 C, anddetector temperature 300 C.

Atherogenic (AI) and thrombogenic indices (TI) were calculatedusing equations described by Ulbricht and Southgate (1991):

AI C12 : 0 4 C14 : 0 C16 : 0=MUFA n 6 n 3 (1)

TI C14 : 0 C16 : 0 C18 : 0=0:5MUFA 0:5 n 6 3 n 3 n 3=n 6 (2)

In the equations, the 14:0 fatty acid is considered to be fourtimes more atherogenic than the other fatty acids; thus a coef-cient of 4 has been assigned to it. Polyunsaturated fatty acids(PUFAs) n6 and monounsaturated fatty acids (MUFAs) have beenassigned coefcients of 0.5 since they are less anti-atherogenic thanPUFAs n3, which have been assigned a coefcient of 3 (Pikul et al.,2008).

Analysis of fat-soluble vitamins A (a-retinol), D3 (chol-cecalciferol), E (a-tocopherol), K2 (menanchion) and b-carotenewas carried out using the method described by Puppel et al.(2012a). The content of cholesterol was determined as describedby Strza1kowska, Jzwik, Bagnicka, Krzy _zewski, and Horbanczuk(2009).

before absorption in small intestine, and a very limited absorptionof both CLA and trans-vaccenic acid from the large intestine of thehorse (Salimei & Fantuz, 2012).

Similar amounts of SFAs andMUFAs in late lactation mares milkwere found by Pikul and Wjtowski (2008) and Pikul et al. (2008).The content of PUFAs in the examined mares milk made up 21% oftotal fatty acids. Similar results in early lactation mares milk wereobtained byMarconi and Panli (1998) and by Pikul andWjtowski(2008) in the milk from the 5th month of lactation. However, inexperiments conducted by Csap et al. (1995), Pietrzak-Fiecko et al.(2009) and Pikul et al. (2008), the content of PUFAs in early and latelactation mares milk was higher and ranged from 28 to 36%. Itneeds to be stressed that the comparison of total SFAs, MUFAs and

rnati3. Results and discussion

3.1. Fat and cholesterol content and fatty acid prole of latelactation mares milk

Late lactation mares milk was characterised by low fat andcholesterol content in comparison with ruminant or human milk(Table 1). Presented results were similar to those recorded by otherauthors and proved that mares milk is a low-fat and low-cholesterol product and differs from cows milk (Martuzzi,Summer, Formaggioni, & Mariani, 2004; Pikul & Wjtowski,2008; Strza1kowska et al., 2009).

Dietary lipids provide essential fatty acids and facilitate theabsorption of fat-soluble vitamins. However individual saturatedfatty acids (SFAs) have different effects on the concentration of theplasma lipoprotein cholesterol fraction. For example lauric (C12:0),myristic (C14:0) and palmitic (C16:0) acids increase LDL cholesterolwhereas stearic acid (C18:0) has no effect (FAO, 2010). Moreover,dietary n3 and n6 fatty acids play a role in modulating immunefunctions and inammatory processes.

According to Berquin, Edwards, and Chen (2008) the diet ofcontemporary people is characterised by too high proportion ofn6 fatty acids and too low proportion of n3 fatty acids. Increasedlevels of n3 fatty acids in the diet supports prevention andtreatment of cancers, heart diseases, thrombosis, arterial hyper-tension, hyperlipidaemia, senile dementia, Alzheimers disease,depression and rheumatoid arthritis (Burdge & Calder, 2006;Calder, 2009; Carlson, 2001; Chavarro et al., 2007; Cuthbertson,1999; Gago-Dominguez, Yuan, Sun, Lee, & Yu, 2003; Horrobin,2000; Innis, 2007; Simopoulos, 2002).

The fatty acid composition of late lactation mares milk ischaracterised by a low proportion of SFA C14:0, C16:0 and C18:0and a high proportion of C8:0, C10:0 and C12:0 when comparedwith cows milk (Table 1). Similar amounts of C8:0 and C14:0 fattyacids in late lactation mares milk were found by Pikul andWjtowski (2008) and Pikul et al. (2008); however, these authorsfound higher concentration of C12:0 (above 6%) and lower con-centration of C18:0 (0.9e1.15%) in milk obtained in the 5th and 6thmonths of lactation. Relatively low levels of stearic acid (C18:0) inmares milk can be explained by dietary factors and the D9desaturase activity in the equine mammary gland (Salimei &Fantuz, 2012).

The amount of main SFA e C16:0 was similar as compared withstudies conducted by Pikul and Wjtowski (2008) and Pikul et al.(2008), who found 20e23% of this fatty acid in late lactation ma-res milk. Other studies conducted on early lactation mares milkalso report that the content of C16:0 in mares milk uctuatesaround 20%within a range of 18e23% (Csap et al., 1995; Marconi &Panli, 1998; Pietrzak-Fiecko et al., 2009; Pikul & Wjtowski, 2008;Pikul et al., 2008; Rutkowska et al., 2011; Salimei & Fantuz, 2012).This is similar to human milk and lower than in cows milk (Molt-Puigmart, Permanyer, Castellote, & Lpez-Sabater, 2011;ODonnell-Megaro, Barbano, & Bauman, 2011).

In the group of MUFAs, which made up 33.59% of total FA, oleicacid (C18:1c9) was dominant (Table 1). Its concentration in thestudied milk was higher than that presented by Pikul et al. (2008)who found 18% of C18:1c9, and Pikul and Wjtowski (2008) whofound 21% of this fatty acid; although a wide variability in con-centration of this FA can be observed among experimental data.Csap et al. (1995) found only 14%, and Pikul et al. (2008) 16% ofC18:1c9 in the milk from the rst two months of lactation; how-ever, other studies on early lactation mares milk report highervalues of this fatty acid ranging from 20 to 22% (Marconi & Panli,1998; Pikul & Wjtowski, 2008; Rutkowska et al., 2011). The rela-

M. Markiewicz-Keszycka et al. / Intetively high level of C16:1 (5.5%) and C18:1 (25%) found in thecurrent study reects high D9 desaturase activity (Uniacke-Lowe,2011).

PUFAs constituted a considerable proportion of FA in latelactation mares milk, of which C18:2n6 (LA) and C18:3n3 (ALA)were dominant (Table 1). The ratio of LA/ALA in mares milk wasdifferent when compared with milk from ruminants (Jzwik,Strza1kowska, Bagnicka, Po1awska, & Horbanczuk, 2010;Kuczynska, 2011; Markiewicz-Keszycka, Czy _zak-Runowska,Lipinska, & Wjtowski, 2013b; Szumacher-Strabel et al., 2011).Pikul et al. (2008) achieved a similar result for the LA/ALA ratioduring a 6 month lactation period (0.29 and 0.39 for the 5th and6th months of lactation, respectively). The LA/ALA ratio inexperiment of Csap et al. (1995), on early lactation mares milk,was also below 0.5. In turn Pikul and Wjtowski (2008), during 5months of lactation, recorded a higher value for the LA/ALA ratio,ranging from 3.5 in the 1st month of lactation to 2.0 in the 5thmonth of lactation.

These variations in LA/ALA ratios between studies are probablydue to the differences in the fatty acid composition of diets. Rationsricher in forages have more ALA than rations rich in concentrates.Because PUFAs consumed by horses are not microbiologicallydehydrogenated before intestinal absorption, milk long-chain fattyacid composition is related to the fatty acid prole of feedstuffs(Doreau, Boulot, Bauchart, Barlet, & Martin-Rosset, 1992; Salimei &Fantuz, 2012).

Mares milk has exceptional values of AI and TI. Our results for AIand TI were very similar to those obtained by Pikul et al. (2008) inlate lactation mares milk who noted values of 1.01 and 0.37 for AIand TI, respectively. According to data presented by Molt-Puigmart et al. (2011), human milk has AI and TI values similarto the values obtained in the current study. Cows milk is charac-terised by higher AI and TI values when compared with mares andhuman milk (Table 1).

CLA was not detected in the examined mares milk. Very smallamounts of CLA (0.03e0.14% of fatty acids) were found by Pikul andWjtowski (2008), Pikul et al. (2008) and Rutkowska et al. (2011).The difference between the CLA level in ruminant and equine milkis due to the lack of biohydrogenation of n6 octadecadienoic acid

Table 2Fat-soluble vitamins and b-carotene concentration in mare milk from the late stageof lactation in comparison with cow and human milk.

Vitamins Mean (SD) Minimum Maximum Cowa Humanb

A (mg L1) 0.403 (0.081) 0.210 0.515 0.435e0.799 0.455D3 (mg L1) 4.93 (1.63) 2.25 8.89 2.31e15.39 0.03e0.12E (mg L1) 1.13 (0.27) 0.61 1.61 1.05e1.95 5.09K2 (mg L1) 17.39 (7.54) 9.48 32.60 4.81e17 1.80b-carotene

(mg L1)0.388 (0.102) 0.166 0.599 0.166e0.380 0.002e0.375

a Kuczynska et al. (2012); Kuczynska (2011); Puppel et al. (2012a).b Kamao et al. (2007).

onal Dairy Journal 38 (2014) 31e36 33PUFAs between the studies should be done carefully. While Pikul

natiet al. (2008) analysed 16 FA, Pikul and Wjtowski (2008) analysed22, Pietrzak-Fiecko et al. (2009) analysed 18 and in the currentstudy we analysed 13 FA.

Differences in the fatty acid prole between the studies areprobably due to the fact that fatty acid composition is mostamenable to change, in comparison with other milk constituents.Studies on the effects of feeding, breed, stage of lactation or geneticvariation on the fatty acid prole of mares milk are scare. Pikul andWjtowski (2008) and Pikul et al. (2008) found that the FA prole isaffected by stage of lactation; results from the studies of Doreauet al. (1992), conrm that the FA prole depends on feeding; inturn Pietrzak-Fiecko et al. (2009), revealed that FA composition ofmares milk was breed-specic.

3.2. Vitamin A, D, E and K content and b-carotene content of latelactation mares milk

So far, few studies on the fat-soluble vitamins concentration inmares milk have been published. The only available sources ofliterature on this subject include the papers of Csap et al. (1995)and Marconi and Panli (1998).

Table 2 presents the results of analysis for fat-soluble vitamincomposition in milk of CBHs mares in the late stage of lactation.Among fat-soluble vitamins, the most abundantly representedwere vitamins E and A. The least abundant were vitamins D3(0.005 mg L1) and K2 (0.017 mg L1). Vitamin K2 contents inexamined milk were highly variable from the lowest value of 0.009to the highest of 0.033 mg L1. Its concentration in late lactationmares milk examined in this study, was lower compared with theconcentration of vitamin K3 determined by Csap et al. (1995) inearly lactation mares milk (0.029 mg L1). Vitamin K is commonlycalled the antibleeding factor. Apart from the synthesis of bloodclotting factors, one of its biological functions is to participate inbone formation (Morrissey & Hill, 2009).

Vitamin E is a major antioxidant that quenches singlet oxygenand inhibits the formation of peroxide radicals, and thus reducesthe level of DNA damage (Claycombe & Meydani, 2001; Sacheck,Milbury, Cannon, Roubenoff, & Blumberg, 2003). The results ob-tained showed that concentration of vitamin E varied from 0.61 to1.61 mg L1. The most extensive study on this topic, conducted byCsap et al. (1995), conrmed that the concentration of vitamin E inmares milk remains within that range. The above authorsdemonstrated that in the milk collected between d 8 and 45 oflactation, the average vitamin E content was the same as in thisstudy (1.13 mg L1).

Marconi and Panli (1998) determined the vitamin content inpowdered mares milk. In their experiment, fresh milk was ob-tained only once from one mare in the sixth week of lactation. Thelevel of vitamin E determined by these researchers indicated thatvitamin E content in mares milk was lower (0.26 mg L1) than thatobtained in our study and the study of Csap et al. (1995).

Literature data indicate that the concentration of vitamin E incows milk is similar to that obtained in this study for mares milk.The results of the study of Lindqvist, Nadeau, Waller, Jensen, andJohansson (2011) demonstrated that, in the case of cows milk,vitamin E concentration depends on feeding system and season andranges from 0.78 to 1.80 mg L1. Human milk contains morevitamin E at the beginning than at the end of lactation with theconcentration level ranging from 0.9 to 16.6 mg L1 (Kamao et al.,2007; Molt - Puimarti et al., 2011; Ortega, Lpez-Sobaler,Martinez, Andres, & Quintas, 1999; Romeu-Nadal, Castellote, &Lpez-Sabater, 2008; Schweigert, Bathe, Chen, Bscher, & Duden-hausen, 2004).

Other antioxidants in milk are vitamin A and b-carotene, which

M. Markiewicz-Keszycka et al. / Inter34are also required for many biological processes (Dewanto, Wu,Adom, & Liu, 2002; Di Vaio, Graziani, Marra, Cascone, & Ritieni,2008; Sauvant, Grolier, & Azais-Braesco, 2002; Zanni, Corbini, LaRosa, & Dreassi, 2010). For the human organism to be able to utilisecarotenoids as provitamin A, they have to be consumed with acertain amount of fat and protein (Rodriguez-Amaya, 2010).

Milk and milk products are an important source of vitamin A,which is present in the form of trans-retinol and provitamin A-b-carotene. In our study the level of vitamin A in the late lactationmares milk varied from 0.21 to 0.51 mg L1 Csap et al. (1995)obtained similar amounts of vitamin A in early lactation maresmilk (0.34 mg L1). In turn, in study of Marconi and Panli (1998),vitamin A concentration in early lactation mares milk was muchlower (0.09 mg L1) than that obtained in our experiment.

Concentration of total trans-retinol in cows milk is generallysimilar to that obtained in our study for mares milk and rangesfrom 0.29 to 0.42 mg L1 (Johansson, Persson Waller, Jensen,Lindqvist, & Nadeau, 2014). Human milk contains more vitamin Athan cows and mares milk, with values ranging from 0.26 to1.03 mg L1 (Darragh, 2002; Kamao et al., 2007; Mello-Neto et al.,2009; Schweigert et al., 2004).

The results recorded by Marconi and Panli (1998) for b-caro-tene concentration in early lactation mares milk, were much lower(0.03 mg L1) than those obtained in this study. According to Fallaand Emanuelson (2011), Lindqvist et al. (2011) and Nozire et al.(2006), b-carotene content in cows milk is similar to that ob-tained in this study for late lactation mares milk and varies from0.07 to 0.21 mg L1. The factors affecting b-carotene concentrationin cows milk include breed, age, stage of lactation and nutrition(Morrissey & Hill, 2009). To date there have been no reportsavailable on the inuence of these factors on b-carotene level inmares milk.

While in cows milk b-carotene represents 95% of total carot-enoids, in human milk, wide variety of carotenoids are present andb-carotene constitutes only about 20% of all carotenoids (Hulshof,van Roekel-Jansen, van de Bovenkamp, & West, 2006; Kamaoet al., 2007; Schweigert et al., 2004). To the best of our knowledge,the carotenoid prole of mares milk has not been assayed andrequires further research.

Vitamin D is another fat-soluble vitamin that was found insignicant amount in late lactation mares milk (2.25e8.89 mg L1).Csap et al. (1995) obtained a slightly lower average value forvitamin D concentration (3.2 mg L1) in early lactation mares milkthan in our study. The vitamin D content of cows milk is highlyvariable and depends on the vitamin status of the cow, feedingregime as well as on sunlight exposure (Kuczynska, 2011;Kuczynska et al., 2012; Kurmann & Indyk, 1994; Puppel et al.,2012a). According to Oberhelman et al. (2013), human milk usu-ally contains less vitamin D than does infant formula with theconcentrations below 7 ng mL1.

The importance of adequate vitamin D intake for bone health iswell known. Moreover, vitamin D may have other roles in pro-moting health and preventing many diseases. According to Glade(2012), supplementation with vitamin D signicantly decreasedthe risk of premature death and death from cancer, as well assupporting general health.

4. Conclusions

The results from this study conrmed that the fat content of thelate lactation mares milk is remarkably lower than the fat contentof human and ruminant milk. Like human, and unlike cows milk,mares milk is characterised by a low proportion of saturated fattyacids and a relatively high level of MUFAs and PUFAs. The fatty acidprole and fat-soluble vitamin content, indicate that mares milk

onal Dairy Journal 38 (2014) 31e36may constitute a valuable source of these components in the

Jzwik, A., Strza1kowska, N., Bagnicka, E., Po1awska, E., & Horbanczuk, J. O. (2010).The effect of feeding linseed cake on milk yield and fatty acid prole in goats.Animal Science Papers and Reports, 28, 245e251.

_

rnatiKamao, M., Tsugawa, N., Suhara, Y., Wadab, A., Moric, T., Muratad, K., et al. (2007).Quantication of fat-soluble vitamins in human breast milk by liquid chro-matographyetandem mass spectrometry. Journal of Chromatography B, 859,192e200.

Koletzko, B., Rodriguez-Palmero, M., Demmelmair, H., Fidler, N., Jensen, R., &Sauerwald, T. (2001). Physiological aspects of human milk lipids. Early HumanDevelopment, 65, S3eS18.

Koletzko, B., Agostoni, C., Bergmann, R., Ritzenthaler, K., & Shamir, R. (2011).human diet, and may have cosmetic purpose. However, there is stillneed for in-depth knowledge on sustainable production systemsand the use of mares milk to obtain a safe, eco-efcient andcompetitive product, which could meet the rising demand for milk-delivered bioactive lipid components with special consideration oftheir effects on human health.

References

AOAC. (1990). Ofcial methods of analysis. Washington, DC, USA: Association ofOfcial Analysis Chemists.

Arsic, A., Prekajski, N., Vucic, V., Tepsic, J., Popovic, T., Vrvic, M., et al. (2009). Milk inhuman nutrition: comparison of fatty acid proles. Acta Veterinaria, 59, 569e578.

Berquin, I. M., Edwards, I. J., & Chen, Y. Q. (2008). Multi-targeted therapy of cancerby omega-3 fatty acids. Cancer Letters, 269, 363e377.

Burdge, G. C., & Calder, P. C. (2006). Dietary D-linolenic acid and health-relatedoutcomes: a metabolic perspective. Nutrition Research Reviews, 19, 26e53.

Calder, P. C. (2009). Polyunsaturated fatty acids and inammation: therapeuticpotential in rheumatoid arthritis. Current Rheumatology Reviews, 5, 214e225.

Carlson, S. E. (2001). Docosahexaenoic acid and arachidonic acid in infant devel-opment. Seminars in Neonatology, 6, 437e449.

Chavarro, J. E., Stampfer, M. J., Li, H., Campos, H., Kurth, T., & Ma, J. (2007).A prospective study of polyunsaturated fatty acid levels in blood and prostatecancer risk. Cancer Epidemiology, Biomarkers and Prevention, 16, 1364e1370.

Claycombe, K. J., & Meydani, S. N. (2001). Vitamin E and genome stability. MutationResearch, 475, 37e44.

Csap, J., Steer, J., Martin, T. G., Makray, S., & Csap-Kiss, Zs (1995). Composition ofmares colostrum and milk. Fat content, fatty acid composition and vitamincontent. International Dairy Journal, 5, 393e402.

Cuthbertson, W. F. J. (1999). Evolution of infant nutrition. British Journal of Nutrition,81, 359e371.

Darragh, A. (2002). Human milk. In H. Roginski, J. W. Fuquay, & P. F. Fox (Eds.),Encyclopedia of dairy sciences (pp. 1350e1360). San Diego, CA, USA: AcademicPress.

Dewanto, V., Wu, X., Adom, K. K., & Liu, R. H. (2002). Thermal processing enhancesthe nutritional value of tomatoes by increasing total antioxidant activity. Journalof Agricultural and Food Chemistry, 50, 3010e3014.

Di Vaio, C., Graziani, G., Marra, L., Cascone, A., & Ritieni, A. (2008). Antioxidantcapacities, carotenoids and polyphenols evaluation of fresh and refrigeratedpeach and nectarine cultivars from Italy. European Food Research and Technol-ogy, 227, 1225e1231.

Doreau, M., Boulot, S., Bauchart, D., Barlet, J.-P., & Martin-Rosset, W. (1992).Volontary intake, milk production and plasma metabolites in nursing mares fedtwo different diets. Journal of Nutrition, 122, 992e999.

EN ISO 5509. (2000). Animal and vegetable fats and oils- preparation of methyl estersof fatty acid. International Standard ISO 5509:2000. Geneva, Switzerland: In-ternational Standardisation Organisation.

Falla, N., & Emanuelson, U. (2011). Fatty acid content, vitamins and selenium in bulktank milk from organic and conventional Swedish dairy herds during the in-door season. Journal of Dairy Research, 78, 287e292.

FAO. (2010). Fats and fatty acids in human nutrition. Report of an expert consultation.FAO Food and Nutrition Paper, 91. Geneva, Switzerland: WHO/FAO.

Gago-Dominguez, M., Yuan, J. M., Sun, C. L., Lee, H. P., & Yu, M. C. (2003). Opposingeffects of dietary n3 and n6 fatty acids on mammary carcinogenesis: theSingapore Chinese health study. British Journal of Cancer, 89, 1686e1692.

Glade, M. J. (2012). A 21st century evaluation of the safety of oral vitamin D.Nutrition, 28, 344e356.

Horrobin, D. F. (2000). Essential fatty acid metabolism and its modication in atopiceczema. American Journal of Clinical Nutrition, 71, 367Se372S.

Hulshof, P. J. M., van Roekel-Jansen, T., van de Bovenkamp, P., & West, C. E. (2006).Variation in retinol and carotenoid content of milk and milk products in TheNetherlands. Journal of Food Composition and Analysis, 19, 67e75.

Innis, S. (2007). Fatty acids and early human development. Early Human Develop-ment, 83, 761e766.

Johansson, B., Persson Waller, K., Jensen, S. K., Lindqvist, H., & Nadeau, E. (2014).Status of vitamins E and A and b-carotene and health in organic dairy cows feda diet without synthetic vitamins. Journal of Dairy Science, 97, 1682e1692.

M. Markiewicz-Keszycka et al. / IntePhysiological aspects of human milk lipids and implications for infant feeding: aworkshop report. Acta Paediatrica, 100, 1405e1415.Kuczynska, B. (2011). Bioactive components and technological parameters of milkproduced at ecological and conventional farms. Dissertations and Monographs.Warsaw, Poland: SGGW (in Polish, abstract in English).

Kuczynska, B., Puppel, K., Go1ebiewski, M., Kordyasz, M., Grodzki, H., &Brzozowski, P. (2012). Comparison of fat and protein fractions of milk constit-uents in Montbeliarde and Polish Holstein-Friesian cows from one farm inPoland. Acta Veterinaria Brno, 81, 139e144.

Kurmann, A., & Indyk, H. (1994). The endogenous vitamin D content of bovine milk:inuence of season. Food Chemistry, 50, 75e81.

Lindqvist, H., Nadeau, E., Waller, K.-P., Jensen, S., & Johansson, B. (2011). Effects ofRRR-a-tocopheryl acetate supplementation during the transition period onvitamin status in blood and milk of organic dairy cows during lactation. Live-stock Science, 142, 155e163.

Marconi, E., & Panli, G. (1998). Chemical composition and nutritional properties ofcommercial products of mare milk powder. Journal of Food Composition andAnalysis, 11, 178e187.

Markiewicz-Keszycka, M., Wjtowski, J., Kuczynska, B., Puppel, K., Czy _zak-Runowska, G., Bagnicka, E., et al. (2013a). Chemical composition and wheyprotein fraction of late lactation mares milk. International Dairy Journal, 31, 62e64.

Markiewicz-Keszycka, M., Czy _zak-Runowska, G., Lipinska, P., & Wjtowski, J.(2013b). Fatty acid prole of milk e a review. Bulletin of the Veterinary Institutein Pulawy, 57, 135e139.

Martuzzi, F., Summer, A., Formaggioni, P., & Mariani, P. (2004). Milk of Italian Saddleand Hainger nursing mares: physico-chemical characteristics, nitrogencomposition and mineral elements at the end of lactation. Italian Journal ofAnimal Science, 3, 293e299.

Mello-Neto, J., Rond, P. H. C., Oshiiwa, M., Morgano, M. A., Zacari, C. Z., &Domingues, S. (2009). The inuence of maternal factors on the concentration ofvitamin A in mature breast milk. Clinical Nutrition, 28, 178e181.

Moate, P. J., Chalupa, W., Boston, R. C., & Lean, I. J. (2007). Milk fatty acids. I. Vari-ation in the concentration of individual fatty acids in bovine milk. Journal ofDairy Science, 90, 4730e4739.

Molt-Puigmart, C., Permanyer, M., Castellote, A., & Lpez-Sabater, M. (2011). Ef-fects of pasteurisation and high-pressure processing on vitamin C, tocopherolsand fatty acids in mature human milk. Food Chemistry, 124, 697e702.

Morrissey, P., & Hill, T. (2009). Fat-soluble vitamins and vitamin C in milk and milkproducts. In P. L. H. McSweeney, & P. F. Fox (Eds.), Advanced dairy chemistry (pp.527e589). New York, NY, USA: Springer.

Nozire, P., Grolier, P., Durand, D., Ferlay, A., Pradel, P., & Martin, B. (2006). Variationsin carotenoids, fat-soluble micronutrients, and color in cows plasma and milkfollowing changes in forage and feeding level. Journal of Dairy Science, 89,2634e2648.

Oberhelman, S. S., Meekins, M. E., Fischer, P. R., Lee, B. R., Singh, R. J., Cha, S. S., et al.(2013). Maternal vitamin D supplementation to improve the vitamin D status ofbreast-fed infants: a randomized controlled trial. Mayo Clinic Proceedings, 88,1378e1387.

ODonnell-Megaro, A., Barbano, D., & Bauman, D. (2011). Survey of the fatty acidcomposition of retail milk in the United States including regional and seasonalvariations. Journal of Dairy Science, 94, 59e65.

Ortega, R. M., Lpez-Sobaler, A. M., Martinez, R. M., Andres, P., & Quintas, M. E.(1999). Inuence of smoking on vitamin E status during the third trimester ofpregnancy and on breast-milk tocopherol concentrations in Spanish women.American Journal of Clinical Nutrition, 68, 662e667.

Palupi, E., Jayanegara, A., Ploegera, A., & Kahla, J. (2012). Comparison of nutritionalquality between conventional and organic dairy products: a meta-analysis.Journal of the Science of Food and Agriculture, 92, 2774e2781.

Pietrzak-Fiecko, R., Tomczynski, R., Swistowska, A., Borejszo, Z., Kokoszko, E., &Smoczynska, K. (2009). Effect of mares breed on the fatty acid composition ofmilk fat. Czech Journal Animal Science, 54, 403e407.

Pikul, J., & Wjtowski, J. (2008). Fat and cholesterol content and fatty acidcomposition of mares colostrum and milk during ve lactation months. Live-stock Science, 113, 285e290.

Pikul, J., Wjtowski, J., Dankw, R., Kuczynska, B., & Lojek, J. (2008). Fat content andfatty acids prole of colostrum and milk of primitive Konik horses (Equuscaballus gmelini Ant.) during six months of lactation. Journal of Dairy Research,75, 302e309.

Puppel, K., Na1ecz-Tarwacka, T., Kuczynska, B., Go1ebiewski, M., Kordyasz, M., &Grodzki, H. (2012a). The age of cows as a factor shaping the antioxidant levelduring a nutritional experiment with sh oil and linseed supplementation forincreasing the antioxidant value of milk. Journal of the Science of Food andAgriculture, 92, 2494e2499.

Puppel, K., Na1ecz-Tarwacka, T., Kuczynska, B., Go1ebiewski, M., & Grodzki, H.(2012b). Inuence of combined supplementation of cows diet with linseed andsh oil on the content of thrombogenic and atherogenic indicators of milk fat.Animal Science Papers and Reports, 30, 317e328.

Rodriguez-Amaya, D. B. (2010). Quantitative analysis, in vitro assessment ofbioavailability and antioxidant activity of food previous carotenoids - a review.Journal of Food Composition and Analysis, 23, 726e740.

Romeu-Nadal, M., Castellote, A. I., & Lpez-Sabater, M. C. (2008). Effect of coldstorage on vitamins C and E and fatty acids in human milk. Food Chemistry, 106,65e70.

Rutkowska, J., Adamska, A., & Bia1ek, M. (2011). Comparison of fatty acid compo-

onal Dairy Journal 38 (2014) 31e36 35sition in mares and cows milk fat. Zywnosc Nauka Technologia Jakosc, 74, 28e38 (in Polish, abstract in English).

Sacheck, J. M., Milbury, P. E., Cannon, J. G., Roubenoff, R., & Blumberg, J. B. (2003).Effect of vitamin E and eccentric exercise on selected biomarkers of oxidativestress in young and elderly men. Free Radical Biology and Medicine, 34, 1575e1588.

Salimei, E., & Fantuz, F. (2012). Equid milk for human consumption. InternationalDairy Journal, 24, 130e142.

Sauvant, P., Grolier, P., & Azais-Braesco, V. (2002). Vitamin A, nutritional signi-cance. In H. Roginski, J. W. Fuquay, & P. F. Fox (Eds.), Encyclopedia of dairy sci-ences (pp. 2657e2664). San Diego, CA, USA: Academic Press.

Schweigert, F. J., Bathe, K., Chen, F., Bscher, U., & Dudenhausen, J. W. (2004). Effectof the stage of lactation in humans on carotenoid levels in milk, blood plasmaand plasma lipoprotein fractions. European Journal of Nutrition, 43, 39e44.

Simopoulos, A. P. (2002). The importance of the ratio of omega-6/omega-3 essentialfatty acids. Biomedical Pharmacotherapy, 56, 365e379.

Strza1kowska, N., Jzwik, A., Bagnicka, E., Krzy _zewski, J., & Horbanczuk, J. O. (2009).Studies upon genetic and environmental factors affecting the cholesterol con-tent of cow milk. I. Relationship between the polymorphic form of beta-

lactoglobulin, somatic cell count, cow age and stage of lactation and choles-terol content of milk. Animal Science Papers and Reports, 27, 95e103.

Szumacher-Strabel, M., Cieslak, A., Zmora, P., Pers-Kamczyc, E., Bielinska, S.,Stanisz, M., et al. (2011). Camelina sativa cake improved unsaturated fatty acidsin ewes milk. Journal of the Science of Food and Agriculture, 91, 2031e2037.

Ulbricht, T., & Southgate, D. (1991). Coronary heart disease: seven dietary factors.Lancet, 338, 985e992.

Uniacke-Lowe, T. (2011). Studies on equine milk and comparative studies on equineand bovine milk systems. PhD Thesis. Ireland: University College Cork.

Uniacke-Lowe, T., Huppertz, T., & Fox, P. F. (2010). Equine milk proteins: chemistry,structure and nutritional signicance. International Dairy Journal, 20, 609e629.

Wan, Z.-X., Wang, X.-L., Xu, L., Geng, Q., & Zhang, Y. (2009). Lipid content and fattyacids composition of mature human milk in rural North China. British Journal ofNutrition, 103, 913e916.

Zanni, A., Corbini, G., La Rosa, C., & Dreassi, E. (2010). Antioxidant activity of tomatolipophilic extracts and interactions between previous carotenoids and a-tocopherol in synthetic mixtures. LWT e Food Science and Technology, 43, 67e72.

M. Markiewicz-Keszycka et al. / International Dairy Journal 38 (2014) 31e3636

Concentration of selected fatty acids, fat-soluble vitamins and -carotene in late lactation mares' milk1 Introduction2 Materials and methods2.1 Animals, feeding and milk sample collection2.2 Experimental procedures

3 Results and discussion3.1 Fat and cholesterol content and fatty acid profile of late lactation mares' milk3.2 Vitamin A, D, E and K content and -carotene content of late lactation mares' milk

4 ConclusionsReferences