Small Ruminant Research 226 (2023) 107058

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Small Ruminant Research

journal homepage: www.elsevier.com/locate/smallrumres

Influence of marine algae (Schizochytrium sp.) supplementation, ripening

and vacuum packaging on goat cheese composition and fatty acid profile
Ákos Bodnár a, István Egerszegi a, Péter Póti a, Jan Kuchtik b, Ferenc Pajor a, *
a
Hungarian University of Agriculture and Life Sciences, Institute of Animal Sciences, Department of Animal Husbandry Technology and Animal Welfare, Páter Károly 1,
H-2100 Gödöllő, Hungary
b
Mendel University in Brno, Faculty of AgriSciences, Zemědělská 1665/1, CZ-61300 Brno, Czech Republic

A R T I C L E I N F O A B S T R A C T

Keywords: The effect of diet (marine algae – MA; control – C), time of ripening (2 days, 4 and 8 weeks) and vacuum
Cheese packaging on the composition and fatty acid (FA) profile of semi-hard cheese were evaluated in this study.
Maturation Twenty-eight Alpine goats were housed indoors and randomly allocated to two groups: control group (C) was fed
DHA
by 1500 g alfalfa hay and 600 g concentrate (n = 14); experimental group (MA) was fed by the same feed as C
Vacuum folie
Beneficial effect
group supplemented 10 g/head/day Schizochytrium limacinum (n = 14). The study lasted 45 days, when milk
samples were taken the last 10 days for their processing and analysis. Cheese samples (C: n = 15; MA: n = 25)
were prepared from bulk milk from both groups and ripened. On day 2 of ripening, ten MA cheese samples were
vacuum packed in foil. The semi-hard cheese samples were collected at 2nd day, then 4th and 8th weeks of
ripening period. The MA supplementation did not significantly affect the composition of the milk. However, the
MA supplementation had a significant (p < 0.05) effect on chemical composition of cheese, with the exception of
fat content and fat content in TS. The supplementation of MA significantly (p < 0.05) increased the contents of
DHA and rumenic acid in the cheese samples (MA group: 0.18 and 0.34 g/100 g, C group: 0.0 and 0.30 g/100 g of
fatty acids (FA), respectively). The MA supplementation had a significant (p < 0.05) effect on contents of all odd-
and branched-chain fatty acids in cheese, with the exception of C9:0. The ripening had significant (p < 0.05)
influence on the rumenic acid (c9t11 C18:2) and alpha-linolenic acid (C18:3) content in the cheese. Vacuum
packaging (VP) had a significant (p < 0.05) effect on the contents of fat, fat in TS and TS in cheese, while their
contents were in all cases higher in unpackaged cheese. The VP also had a significant (p < 0.05) effect on the
contents of most of monitored FAs in cheese.

1. Introduction
The total world production of cheese in 2020 was approximately
more than 21,743 thousand tons, including 511 thousand tons of goat
cheese (FAO, 2023). The consumers’ interesting for goat milk and dairy
products has intensively increased during the last decades due to their
bioactive substances with a beneficial effect on the human health (such
as healthier fatty acid composition). Moreover, goat milk is more ad­
vantageous than cow milk regarding smaller fat globules and softer curd
formation during digestion (Park, 2007). In the period from 1990 to
2020, the total world production of goat milk increased by about 207%,
while the total production of cow milk increased in the same period by
only 155% (FAO, 2023). In addition, the production of goat milk and
dairy products are economically important in several regions and
countries.
Nowadays, some of the fatty acids (FA) are linked with unhealthy
properties, consequently, there has been increased interest in the
modifying of the FA composition of dairy products, such as improve the
concentration of n-3 FAs. The most common way to improve the FA
composition of milk and dairy products with n-3 polyunsaturated fatty
acids (PUFA) is to supplement animal diets with them (Toral et al., 2017;
Pajor et al., 2021). Marine algae supplements are not only good source of
n-3 PUFA, but also contains high concentrations of long-chain n-3 PUFA,
such as docosahexaenoic acid (DHA), whilst DHA has great beneficial
effects for human health, such as reducing the risk of coronary heart
disease (Kerdiles et al., 2017). Marine-originated supplements (such as
fish oil and algae) contain notable DHA concentrations (Toral et al.,
2017). Nevertheless, advantages of marine algae compared to the fish oil

* Corresponding author.
E-mail address: pajor.ferenc@uni-mate.hu (F. Pajor).

https://doi.org/10.1016/j.smallrumres.2023.107058
Received 30 May 2023; Received in revised form 9 July 2023; Accepted 21 July 2023
Available online 24 July 2023
0921-4488/© 2023 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
Á. Bodnár et al.
are faster growth rate and high productivity of DHA (Ren et al., 2010).
The importance of DHA for human health is well known and therefore
the European Food Safety Authority (EFSA) and the United Nations Food
and Agriculture Organization (FAO) recommend it for human con­
sumption. Presently, the recommended daily intake of eicosapentaenoic
acid (EPA) + docosahexaenoic acid (DHA) fatty acids for an adult is 250
mg (EFSA, 2010) and for pregnant and lactating women is 300 mg/day,
but at least 200 mg/day should be DHA (FAO, 2008).
Ripening is a crucial technological process in cheese manufacturing.
Rennet-coagulated cheeses are ripened for periods ranging from about
two weeks to two or more years. The ripening process of cheese is
compounded and includes microbiological and biochemical changes to
the curd resulting in the aroma and texture attribute of the different type
of cheeses due to its effect on the chemical composition (Fox et al., 2015;
Khattab et al., 2019).
Vacuum packaging is the most common method to reduce the water
and weight loss of cheeses (Nájera et al., 2021). During the vacuum
packaging, the air is removed around the cheese, using a suitable
packaging material with low permeability to oxygen (Miloradovic et al.,
2018). Previously, Romani et al. (2002) confirmed the usefulness of
vacuum packaging for preservation and cheese shelf-life extension.
Vacuum packaging changes the biochemical processes in the cheese
during ripening. However, the effect of vacuum packaging on proteol­
ysis and lipolysis during cheese maturing is controversial when for
example Hayaloglu et al. (2012) found more intensive proteolytic and
lipolytic processes during ripening. In contrast, Barać et al., (2021) and
Garabal et al. (2010) reported a lower proteolysis and lipolysis in the
cheese ripened in a vacuum.
Previously, when the daily marine algae supplementation in dairy
ewes and goats’ diets were 10–60 g per head, MA intake has significantly
increased the DHA concentration in milk. However, high daily intake of
MA associated with reduced milk fat content in sheep and goat milk
(Mavrommatis and Tsiplakou, 2020; Pajor et al., 2021; Zisis et al.,
2022).
We hypothesized that daily 10 g per animal marine algae enriched
diet significantly influences the fatty acid profile of cheese samples,
moreover, ripening and vacuum packaging can modify the chemical
composition and fatty acid profile of them.
The present study aimed to evaluate the composition and fatty acid
profile of goat cheese according to marine algae supplementation (10 g/
head/day) and ripening (8 weeks), as well as vacuum packaging on
chemical composition and fatty acid profile of the semi-hard goat
cheeses.
2. Materials and methods
2.1. Experimental design
This study was carried out on an Alpine goat farm in Ősagárd
(Nógrád County, Hungary; coordinates: 47◦ 85′34.96″N 19◦ 18′77.50″E).
During this study, the investigated animals care was in line with the EU
directive on the protection of animals used for scientific purposes.
Twenty-eight Alpine goats (DIM: 161 ± 3.61d) were involved in this
study, which were equalized by parity (2nd and 3rd lactation), time of
kidding (March) and kid rearing (8 w). During the entire experimental
period, goats were kept indoor in a barn with pens. After weaning, all
goats were milked twice a day by machine milking and individually
recorded each day. Goats were randomly allocated to two groups, con­
trol (C) and experimental group (MA), which were balanced by daily
milk yield (control group: 1.26 kg/d; experimental group: 1.27 kg/d,
respectively). The study lasted 45 days, when milk samples were taken
the last 10 days for their processing and analysis. Cheese samples were
prepared from milk from different bulk tanks by groups and ripened.
The animals in the C group were fed by 1500 g alfalfa hay and 600 g
concentrate (ingredients of concentrate, g/kg: maize grain, 300; winter
wheat, 150; soybean hull, 150; sugar beet pellets, 100; sunflower meal,
2
Small Ruminant Research 226 (2023) 107058
100; soybean meal, 70; sodium bicarbonate, 50; calcium carbonate, 50;
salt, 30); in the MA group, goats were fed by the same feed as C group
with addition 10 g/head/d of dried Schizochytrium limacinum marine
algae (ALL-G-RICH®; Dunboyne, Co Meath, Ireland; chemical compo­
sition: dry matter (DM): 929 g/1000 g, crude protein: 148 g/kg DM,
crude fat: 482 g/kg DM, crude fibre: 23 g/kg DM, ash: 38 g/kg DM, DHA:
30.10 g/100 fatty acids). The concentrates in both groups were offered
individually to the animals two times (at morning and evening) during
milking. Alfalfa hay was given in barn to the animals, in two equal parts,
after morning and evening milking. Drinking water was offered ad
libitum to all animals. The chemical composition of diets were iso­
nitrogenous and isoenergetic. The crude protein and net energy for
lactation (NEl) were 198.57 g/kg DM and 198.32 g/kg DM and 6.17 MJ/
kg DM and 6.18 MJ/kg DM for C and MA, respectively. Main fatty acids
(g/100 g of fatty acids) were C14:0 (0.58 and 1.21), C16:0 (13.17 and
17.52), C18:1n-9 (30.33 and 27.53), C18:2n-6 (32.04 and 29.02) and
DHA (0.0 and 2.85) for C and MA, respectively.
2.2. Cheese processing and collect of cheese samples
Cheese samples (~350 g) from MA group (n = 5 per day) were
prepared every other (odd) day (total cheese samples in MA group, n =
25), while those from C group (n = 3 per day) were processed every
other (even) day (total cheese samples in C group, n = 15) from bulk
milk independently during the investigation period. In addition, at 2nd
day ten cheese samples from MA group were packed in vacuum foil
(maximum oxygen transmission rate 50 cm3 m− 2 24 h− 1 1 atm− 1;
maximum water vapour transmission rate 2.3 g m− 2 24 h− 1 1 atm− 1). All
cheese samples were ripening for 8 weeks in a controlled ripening
chamber (temperature: 13 0C, relative humidity: 90%)(Model: STG ALL
700 INOX CF Evertouch (Everlasting s.r.l., Suzzara, Italy). Cheese sam­
ples were collected at day 2, week 4 and week 8, then frozen and stored
at − 20 ◦ C until further analysis. The description of cheese processing is
presented in Table 1.
2.3. Chemical analysis
Fat, protein, lactose, and total solids contents of milk were deter­
mined using an IR spectrometry analyser (LactoScope™, Delta In­
struments, Drachten, The Netherlands). The cheese samples’ fat, protein
and total solids content were analysed as described in the Hungarian
Standards (Hungarian Standard, 1978, 1980 and 2002). Moisture on a
fat-free basis (MFFB, %) was calculated as follows: MFFB =
(100-TS)/(100-Fat content) x 100. Fat in total solids was calculated as
follows: Fat/TS x 100.
The concentrations of individual fatty acids in each cheese samples
were determined by a gas chromatograph method described by Samková
et al. (2020). Cheese fat was extracted with petroleum ether from
freeze-dried cheese samples. Fatty acids in isolated fat were reesterified
Table 1
Description of the cheese processing.
Processing steps Description
Heating of raw milk Temperature: up to 35 ◦ C for 15 min
Added commercial bacterial CHN 11 (mesophilic aromatic culture) and Chy-
culture and rennet to the milk Max commercial rennet (fermentation-
produced chymosin), duration 5 min
Coagulation of milk Up to 60 min
Post heating treatment Temperature: up to 38 ◦ C, duration 20 min
Scooping and draining of Curd cutting; size of pieces ~7 mm (wheat size)
coagulated milk
Formation of cheeses blocks Approx. 1500 g of each
Pressing of cheese blocks 5 kg/kg cheese, duration: 3 h
Salt bath of cheese blocks Salt concentration: 18%, salt temperature:
13 ◦ C, duration: 36 h, temperature: 17–18 ◦ C
Ripening of cheeses Relative humidity: 90%, temperature: 13 ◦ C on
oak board, duration: 8 weeks
Á. Bodnár et al.
to their methyl esters with a methanolic solution of potassium hydrox­
ide. Methyl esters of FA were determined by a gas chromatographic
method (GC) using a Varian 3800 apparatus (Varian Techtron, Palo Alto,
CA, USA) with a flame ionization detector (FID) and 4000 MS detector
on a column (Varian Techtron, 50 m × 0.25 mm, 0.25 µm). The split
injection ratio was 10:1. Helium was used as the carrier gas, applying a
flow rate of 1.8 ml/min. Both the temperature of the injector and de­
tector was 250 ◦ C. The oven temperature programme started at 55 ◦ C for
5 min, then increased 40 ◦ C/min up to 170 ◦ C, and then increased
2 ◦ C/min up to 196 ◦ C, from this point increased again 10.0 ◦ C/min up to
210 ◦ C then 8 ◦ C/min.
The identification of FA was carried out using analytical standards
(Supelco, Darmstadt, Germany). The proportions of individual FA were
calculated from the ratio of their peak area to the total area of all the
observed FA. In total, 50 FA were determined by GC method. The
selected FA combinations were calculated by using FA data: saturated
fatty acids (SFA); monounsaturated fatty acids (MUFA); polyunsaturated
fatty acids (PUFA); OBCFA: odd- and branched-chain fatty acids; total n-
6 and n-3 PUFA and n-6/n-3 ratio.
2.4. Statistical analysis
Statistical analysis using the general linear model (GLM) method was
processed by the SPSS 27.0 software package. Shapiro–Wilk’s test was
used for testing the normality distribution. The effect of diet (C and MA)
on milk composition was assessed by F-test and Student’s t-test.
The effect of diet (C and MA; n = 15 per group), ripening time (2nd
days, 4th and 8th weeks; n = 10 per time), and the interaction of diet and
ripening time on cheese composition and FA profile were determined.
The statistical model was as follows:
yijk = μ + Di + Rj + (D x R)k + eijk
where yijk is the value of the dependent variable, μ is the overall mean,
Di is the effect of diet, Rj is the effect of ripening time, (D x R)k is the
effect of the interaction of diet and ripening time, and eijk is the random
error.
The effect of vacuum packaging (UP – unpackaged and VP – vacuum
packaged, n = 10 per group), ripening time (4th and 8th weeks, n = 10
per time), and the interaction of vacuum packaging and ripening time on
cheese composition and FA profile were also determined. The statistical
model was as follows:
yijk = μ + Pi + Rj + (P x R)k + eijk
where yijk is the value of the dependent variable, μ is the overall mean,
Pi is the effect of vacuum packaging, Rj is the effect of ripening time, (P x
R)k is the effect of the interaction of vacuum packaging and ripening
time, and eijk is the random error.
When the analysis of variance showed significant differences be­
tween the groups, the LSD post hoc test was applied. The differences
were considered significant if p < 0.05.
3. Results
The mean values of milk components are reported in Table 2.
In this study (Table 2), the marine algae supplementation did not
Table 2
Effect of marine algae enriched diet on the goat milk composition.
Traits C (n = 10) MA (n = 10) SEM P-value
Fat% 3.62 3.66 0.057 0.724
Protein, % 3.39 3.37 0.085 0.919
Lactose, % 4.54 4.46 0.058 0.557
Total solids, % 12.28 12.37 0.122 0.732
C-control diet (hay and concentrate), MA-feed as C group supplemented 10 g/
head/day marine algae.
Small Ruminant Research 226 (2023) 107058
significantly affect the composition of the milk. The contents of fat and
protein in milk were 3.62% and 3.66% for C and 3.66% and 3.37% for
MA groups.
The cheese composition is shown in Table 3.
MA supplementation (Table 3) had a significant (p < 0.05) effect on
chemical composition of cheese, with the exception of fat content and fat
content in TS. However, slightly higher contents of fat and fat in TS were
found in the cheese of the group with MA addition.
In contrast, the ripening period had a significant (p < 0.05) effect on
all monitored contents, except content of MFFB, when the contents of
fat, protein, fat in TS and TS were significantly higher after four weeks of
ripening than their contents after two days of ripening. On the other
hand, the contents of all monitored chemical components of the cheese
were comparable after eight weeks of ripening with their values found
after four weeks of ripening. In conclusion to this part, it must be added
that the interaction between diet and periods of ripening had no sig­
nificant effect on all monitored chemical components of cheese.
The fatty acid profiles of cheese samples are presented in Table 4.
The MA supplementation had a significant (p < 0.05) effect on most
of the monitored fatty acids (FA) in cheese. The concentrations of
butyric (C4:0), caproic acid (C6:0), caprylic acid (C8:0), capric acid
(C10:0), lauric acid (C12:0), myristic acid (C14:0), vaccenic acid (t11
C18:1), rumenic acid (c9t11 C18:2), docosahexaenoic acid (C22:6) and
SFA were found to be significantly (p < 0.05) higher in cheese from the
group with MA supplementation.
In contrast, the concentrations of myristoleic acid (C14:1), palmitic
acid (C16:0), palmitoleic acid (C16:1), stearic acid (18:0), oleic acid
(c11 C18:1), linoleic acid (C18:2), alpha-linolenic acid (C18:3), doco­
sapentaenoic acid (C22:5), MUFA, PUFA, n-6 fatty acids, and n-6/n-3
ratio were significantly (p < 0.05) higher in the cheese of the control
group.
(1) The ripening had significant (p < 0.05) influence on the rumenic acid
(c9t11 C18:2) and alpha-linolenic acid (C18:3) content in the cheese.
During ripening, the rumenic acid concentration slightly increased from
0.30 g/100 g of FA at 2nd day to 0.31 g/100 g of FA at the 4th week, then
significantly (P < 0.05) increased to 0.34 g/100 g of FA at the 8th week.
The interaction between diet and ripening had no significant influ­
ence on the FA of cheese fat.
The odd- and branched-chain fatty acid profiles of cheese samples
are presented in Table 5.
In this study, 14 odd- and branched-chain fatty acids were identified
in both groups (Table 5), including 8 odd-chain FA (C9:0, C11:0, C13:0,
(2) C15:0, C15:1, C17:0, C17:1 and C19:1), 6 branched-chain FA (iso-C10,
iso-C14:0, iso-C16:0, iso-C17:0, anteiso-C15:0 and anteiso-C17:0). The
MA supplementation (Table 5) had a significant (p < 0.05) effect on
contents of all odd- and branched-chain fatty acids in cheese, with
exception of C9:0.
The effect of vacuum packaging and ripening on the chemical
composition of the cheese is presented in Table 6.
Vacuum packaging (Table 6) had a significant (p < 0.05) effect on
the contents of fat, fat in TS and TS in cheese, while their contents were
in all cases higher in unpackaged cheese. In contrast, the ripening and
the packaging x ripening interaction had no significant (P < 0.05) effect
on the chemical composition of cheese.
The effect of vacuum packaging and ripening on fatty acid profile of
cheese is presented in Table 7.
The vacuum packaging (Table 7) had a significant (p < 0.05) effect
on the contents of most of monitored FAs in cheese, when the concen­
trations of myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid
(C16:1), stearic acid (C18:0), vaccenic acid (t11 C18:1), linoleic (C18:2),
odd fatty acids, MUFA, PUFA, n-6 fatty acids and n-6/n-3 ratio were
significantly (p < 0.05) higher in vacuum packaged cheese.
On the contrary, the concentrations of butyric (C4:0), caproic acid
(C6:0), caprylic acid (C8:0), capric acid (C10:0), lauric acid (C12:0),
myristoleic acid (C14:1) and saturated fatty acids were significantly (p
< 0.05) higher in unpackaged cheese. The ripening had a significant (p
3
Á. Bodnár et al. Small Ruminant Research 226 (2023) 107058

Table 3
Effect of marine algae enriched diet and ripening on chemical composition of cheese.
Traits Diet (D) Ripening (R) SEM P-value

C (n = 15) MA (n = 15) 2d (n = 10) 4w (n = 10) 8w (n = 10) D R DxR

MFFB1 59.26a 61.54b 61.49 59.99 59.70 0.371 0.005 0.128 0.544
Fat% 21.40 22.20 20.00a 21.20b 22.30b 0.229 0.317 0.002 0.862
Protein, % 21.13b 18.87a 18.60a 20.30b 21.10b 0.312 0.001 0.010 0.863
Fat in TS, % 40.04 40.76 39.38a 40.19a 41.64b 0.278 0.209 0.010 0.339
Total solids, % 53.40b 51.33a 50.80a 52.70b 53.60b 0.384 0.013 0.020 0.793

C-control diet (hay and concentrate), MA-feed as C group supplemented 10 g/head/day marine algae, 1MFFB–moisture content on a fat-free basis; TS-total solids, a,b-
means with different letters differ significantly at p < 0.05.

Table 4
Effect of marine algae enriched diet and ripening on fatty acid profile of cheese (g/100 g of fatty acids).
Fatty Acids Diet (D) Ripening (R) SEM P-value

C (n = 15) MA (n = 15) 2d (n = 10) 4w (n = 10) 8w (n = 10) D R D×R

C4:0 0.75a 1.34bb 1.07 1.02 1.05 0.011 0.000 0.141 0.069
C6:0 1.50a 2.13b 1.84 1.78 1.82 0.013 0.000 0.169 0.014
C8:0 2.18a 2.85b 2.654 2.49 2.51 0.013 0.000 0.309 0.078
C10:0 9.97a 11.22b 10.65 10.52 10.60 0.029 0.000 0.227 0.100
C12:0 5.75a 6.07b 5.91 5.90 5.93 0.008 0.000 0.324 0.255
C14:0 12.47a 13.00b 12.73 12.72 12.76 0.013 0.000 0.307 0.013
C14:1 0.56b 0.52a 0.54 0.54 0.53 0.001 0.000 0.543 0.165
C16:0 28.37b 27.79a 28.07 28.08 28.10 0.037 0.000 0.345 0.012
C16:1 0.37b 0.34a 0.36 0.36 0.36 0.001 0.000 0.627 0.103
C18:0 6.42b 5.68a 6.04 6.06 6.04 0.013 0.000 0.718 0.166
c11 C18:1n-9 17.37b 15.09a 16.2 16.36 16.29 0.033 0.000 0.139 0.457
t11 C18:1n-7 1.24a 1.61b 1.42 1.43 1.42 0.003 0.000 0.193 0.255
rumenic acid 0.30a 0.34b 0.30a 0.31 0.33b 0.006 0.046 0.041 0.002
C18:2n-6 2.54b 2.41a 2.47 2.48 2.47 0.004 0.000 0.258 0.625
C18:3n-3 0.78b 0.63a 0.70 0.71b 0.70a 0.002 0.000 0.018 0.497
C20:4n-6 0.12 0.12 0.11 0.12 0.11 0.001 0.422 0.087 0.822
C20:5n-3 (EPA) 0.04 0.04 0.04 0.04 0.04 0.001 0.536 0.698 0.752
C22:5n-3 0.09b 0.07a 0.08 0.08 0.08 0.001 0.000 0.507 0.328
C22:6n-3 (DHA) 0.00a 0.18b 0.09 0.095 0.089 0.002 0.000 0.201 0.201
OBCFA 5.09b 4.44a 4.75 4.80 4.75 0.017 0.000 0.274 0.738
SFA 72.48a 74.73b 73.60 73.35 73.54 0.044 0.000 0.116 0.629
MUFA 23.20b 21.02a 22.00 22.19 22.02 0.039 0.000 0.115 0.957
PUFA 4.32b 4.25a 4.26 4.30 4.30 0.009 0.000 0.130 0.341
n-6 2.66b 2.52a 2.59 2.6 2.59 0.004 0.000 0.560 0.758
n-3 0.87 0.88 0.87 0.87 0.87 0.004 0.085 0.178 0.307
n-6/n-3 ratio 3.07b 2.87a 2.96 2.95 2.99 0.009 0.000 0.258 0.922

C-control diet (hay and concentrate), MA-feed as C group supplemented 10 g/head/day marine algae; OBCFA-odd- and branched-chain fatty acids; a,b-means with
different letters differ significantly at p < 0.05.

Table 5
Effect of marine algae enriched diet and ripening on odd- and branched-chain fatty acids of cheese (g/100 g of fatty acids).
Fatty Acids Diet (D) Ripening (R) SEM P-value

C (n = 15) MA (n = 15) 2d (n = 10) 4w (n = 10) 8w (n = 10) D R D×R

C9:0 0.09 0.09 0.09 0.09 0.09 0.002 0.946 0.933 0.078
isoC10:0 0.09b 0.08a 0.08a 0.08a 0.09b 0.001 0.022 0.001 0.324
C11:0 0.12b 0.11a 0.12 0.12 0.12 0.001 0.000 0.236 0.005
C13:0 0.10b 0.09a 0.09 0.09 0.09 0.001 0.005 0.814 0.121
isoC14:0 0.18b 0.15a 0.16a 0.17b 0.17b 0.001 0.000 0.000 0.351
C15:0 1.22b 1.10a 1.16 1.16 1.16 0.001 0.000 0.773 0.052
anteisoC15:0 0.53b 0.46a 0.49 0.50 0.49 0.002 0.000 0.131 0.815
C15:1 0.13b 0.11a 0.11 0.13 0.11 0.003 0.003 0.094 0.053
isoC16:0 0.35b 0.27a 0.30a 0.31b 0.31b 0.001 0.000 0.041 0.778
isoC17:0 0.43b 0.39a 0.41 0.41 0.41 0.001 0.000 0.675 0.035
anteisoC17:0 0.56b 0.50a 0.53 0.53 0.53 0.002 0.000 0.850 0.149
C17:0 0.77b 0.66a 0.72 0.72 0.71 0.003 0.000 0.353 0.457
C17:1n7cis 0.39b 0.31a 0.35 0.35 0.35 0.001 0.000 0.605 0.835
C19:1 0.15b 0.11a 0.13 0.14 0.13 0.003 0.000 0.563 0.881
branched FA 1.28b 1.03a 1.14 1.18 1.15 0.006 0.000 0.383 0.922
odd FA 3.81b 3.41a 3.61 3.62 3.60 0.011 0.000 0.176 0.378
OBCFA 5.09b 4.44a 4.75 4.80 4.75 0.017 0.000 0.274 0.738

OBCFA-odd- and branched-chain fatty acids; C-control diet (hay and concentrate), MA-feed as C group supplemented 10 g/head/day marine algae; a,b—means with
different letters differ significantly at p < 0.05.

4
Á. Bodnár et al. Small Ruminant Research 226 (2023) 107058

Table 6
Effect of vacuum packaging and ripening on chemical composition of cheese.
Traits Packaging (P)1 Ripening (R) SEM P-value

UP (n = 10) VP (n = 10) 4w (n = 10) 8w (n = 10) P R PxR

MFFB 61.09 61.44 61.56 60.98 0.338 0.636 0.440 0.440

Fat% 21.60b 19.60a 20.40 20.80 0.342 0.003 0.493 0.998
Protein,% 19.60 19.10 19.05 19.70 0.356 0.517 0.368 0.368
Fat in TS,% 41.46b 37.72a 39.99 40.21 0.518 0.009 0.814 0.814
Total solids,% 52.10b 50.60a 51.00 51.70 0.363 0.046 0.327 0.671
1
vacuum packaging of cheese samples at 2nd day of ripening; MFBB-moisture content on a fat-free basis; UP-unpackaged cheese; VP-vacuum packaged cheese; a,b-
means with different letters differ significantly at p < 0.05.

Table 7
Effect of vacuum packaging and ripening on fatty acid profile of cheese (g/100 g of fatty acids).
Fatty acids Packaging (P) Ripening (R) SEM P-value

UP (n = 10) VP (n = 10) 4w (n = 10) 8w (n = 10) P R PxR

C4:0 1.31b 1.18a 1.23 1.27 0.019 0.015 0.072 0.766

C6:0 2.18b 2.01a 2.07 2.13 0.025 0.004 0.236 0.499
C8:0 2.80b 2.65a 2.70 2.75 0.016 0.017 0.402 0.879
C10:0 11.15b 10.36a 10.67 10.84 0.051 0.001 0.105 0.747
C12:0 6.07b 5.92a 5.98 6.02 0.015 0.001 0.303 0.340
C14:0 13.03b 13.15a 13.06 13.13 0.019 0.006 0.079 0.867
C14:1 0.52b 0.48a 0.52 0.49 0.014 0.313 0.387 0.401
C16:0 27.88a 29.07b 28.40 28.55 0.083 0.001 0.383 0.255
C16:1 0.34a 0.35b 0.35 0.35 0.002 0.788 0.447 0.381
C18:0 5.70a 6.08b 5.90 5.88 0.027 0.001 0.738 0.716
c11 C18:1n-9 15.26a 15.68b 15.61b 15.33a 0.063 0.001 0.046 0.190
t11 C18:1n-7 1.61a 1.70b 1.67 1.65 0.006 0.001 0.066 0.766
rumenic acid 0.34 0.34 0.35 0.34 0.007 0.889 0.268 0.715
C18:2n-6 2.41a 2.48b 2.47 2.43 0.007 0.001 0.273 0.321
C18:3n-3 0.63 0.63 0.64b 0.62a 0.002 0.467 0.001 0.997
C20:4n-6 0.11 0.11 0.12 0.11 0.001 0.820 0.432 0.730
C20:5n-3 (EPA) 0.04 0.04 0.04 0.04 0.001 0.853 0.905 0.795
C22:5n-3 0.07 0.07 0.07 0.07 0.001 0.584 0.542 0.577
C22:6n-3 (DHA) 0.18 0.17 0.18 0.18 0.002 0.085 0.110 0.217
branch FA 0.52 0.53 0.54 0.52 0.003 0.131 0.162 0.715
odd FA 3.91a 3.99b 3.96 3.95 0.007 0.001 0.523 0.187
SFA 74.67b 74.03a 74.11a 74.59b 0.072 0.001 0.004 0.114
MUFA 21.06a 21.62b 21.54b 21.14a 0.064 0.001 0.007 0.097
PUFA 4.27a 4.35b 4.35 4.27 0.013 0.008 0.123 0.627
n-6 2.53a 2.59b 2.58b 2.55a 0.007 0.001 0.018 0.323
n-3 0.88 0.88 0.89b 0.87a 0.005 0.677 0.012 0.626
n-6/n-3 ratio 2.87a 2.96b 2.89 2.94 0.013 0.004 0.109 0.250

UP-unpackaged cheese; VP-vacuum packaged cheese; a,b-means with different letters differ significantly at p < 0.05.

< 0.05) effect only on the concentrations of oleic acid (C18:1), alpha-
linolenic acid (C18:3), SFA, MUFA, n-6 and n-3 fatty acids in cheese
and the interaction between vacuum packaging and ripening had no
significant effect on the contents of FAs in cheese.
4. Discussion
In present study, all monitored milk composition parameters
(Table 2) in both groups were within the normal range for dairy goats
reported by earlier (Park et al., 2006; Kuchtík et al., 2015; Pajor et al.,
2019). In the present study the goats’ milk composition was not affected
by daily 10 g MA feeding. This is in concordance with previous reports,
where daily MA supplementation dose was relatively low, such as 105
g/head/day for dairy cows and 15 g/head/day for dairy goats (Moran
et al., 2018; Pajor et al., 2019).
In contrast to milk composition, the MA feeding significantly (p <
0.05) influenced the contents of MFFB, protein and total solids in the
cheese when the protein and total solids contents in cheese from MA
group were lower, while the MFFB content was higher compared with
the control group. In contrast, there were no significant differences be­
tween C and MA groups in fat and fat in total solids contents in cheese.
Regarding to the chemical composition of the cheese samples, the results
found in our study were very similar to the data reported by Fekadu et al.
(2005) and Pajor et al. (2012). Results of the present study suggest that
MA diet may influence the milk processing properties, however, further
studies are needed to test the effect of MA supplementation on cheese­
making parameters.
Moreover, according to the decision of the European Commission,
different cheeses are categorized according to the moisture content
parameter on a fat-free basis (MFFB), whereby five different groups of
cheeses have been created and identified: soft (MFFB ≥ 68%), semi-soft
(68%> MFFB ≥ 62%), semi-hard (62%>MFFB ≥ 55%), hard (55%
>MFFB ≥47%), and very hard (MFFB< 47%). In present study, cheese
samples could be defined as a semi-hard cheese, based on its moisture
content on a fat-free basis (MFFB, 55 < and >68%)(European Com­
mission, 1997).
The ripening not influenced the content of MFFB in cheese (Table 3).
In contrast, the ripening had a significant (p < 0.05) effect on the con­
tents of fat, fat in total solids, protein and total solids in cheese. These
parameters markedly increased with the progress of ripening, which is in
accordance with the study carried out by Guizani et al. (2006) However,
sharply changes were detected between 2nd day and 4th weeks samples
in fat, protein and total solids contents, while the content of fat in TS
reached the highest values at 8th weeks of ripening. These results sug­
gest that the water losing was faster during the first 4 weeks of ripening,
in turn, it was modest during in the last remained ripening period, which

5
Á. Bodnár et al.
is consistent with the results published by Serhan et al. (2010).
In present study, the MA feeding had a significant (p < 0.05) effect on
the concentrations of most of the monitored FAs in cheese, includes
short-chain fatty acids (C4:0-C10:0). Recently, the short-chain saturated
fatty acids (SCFA) are more interesting for nutritionists. The consump­
tion of these FAs has more beneficial effect on human health, such as
SCFA can exert antimicrobial activities and help to avoid malabsorption
syndrome (Tan et al., 2014; Gómez-Cortés et al., 2018). The SCFAs are
rapidly absorbed in the intestine and transferred directly to liver portal
vein (Gómez-Cortés et al., 2018).
The MA supplementation also increased the concentrations of lauric
acid (C12:0), myristic acid (C14:0) and palmitic acid (C16:0). These
fatty acids, mainly C14:0 and C16:0 fatty acids, had higher concentra­
tions in the cheese of the MA group compared to the control group.
In addition, the concentrations of lauric, myristic and palmitic acids
are most strongly linked with higher blood total cholesterol and LDL
cholesterol concentrations in blood serum (Ulbricht and Southgate,
1991). They called as hypercholesterolemic saturated fatty acids.
Recently, lauric acid would have no impact on cardiovascular health
parameters when supplied in the dairy matrix. Matrix components as
calcium, peptides, phosphorus and the milk fat globule membrane
modify blood lipid responses to saturated fatty acids, such as lauric fatty
acid intake (Thorning et al., 2017). Further evidences have also indi­
cated that myristic acid from milk products has a beneficial effect on
lipid biomarkers by increasing HDL cholesterol and decreasing TAG
levels, without changes in LDL cholesterol (Dabadie et al., 2008).
Currently, only high consumption of palmitic acid (more than 8–10% of
daily energy) is a health concern in terms of cholesterol levels and
cardiovascular disease risk (Legrand and Rioux, 2015).
In present study, the MA-enriched diet significantly (p < 0.05)
decreased the concentrations of stearic acid (C18:0) and oleic acid (c11
C18:1), and increased the concentration of trans vaccenic acid in cheese.
The long-chain PUFAs in the animals’ diet inhibits the PUFAs hydro­
genation to C18:0, this resulted in higher concentrations of in­
termediates, such as trans C18:1 (trans vaccenic acid – TVA) fatty acid
and other biohydrogenation intermediates. Some of these have a nega­
tive effect on the viability of protozoa and bacterial populations in
rumen and caused milk fat depression in dairy animals (Or-Rashid et al.,
2008; Toral et al., 2017; Białek et al., 2018). The TVA is a precursor of
rumenic acid; TVA is converted to rumenic acid by Δ9-desaturase in the
mammary gland (Kuhnt et al., 2011). The rumenic acid has several
favourable effects on human health: anti-inflammatory, anticarcino­
genic, anti-atherosclerotic and anti-obesity (Guillocheau et al., 2019).
The reduced availability of stearic acid influenced the concentration of
oleic acid (c11 C18:1) in cheese, accordingly the concentration of oleic
acid was significantly decreased in MA group.
The concertation of n-3 fatty acids in cheese was slightly higher in
MA group. The most important n-3 fatty acids are docosahexaenoic acid
(DHA; 22:6n-3), eicosapentaenoic acid (EPA; 20:5n-3), and α-linolenic
acid (ALA; 18:3n-3). The concentrations of DHA, EPA and ALA were
higher in MA than C cheeses (0.85 vs. 0.82; P < 0.001, not presented in
Table). These FAs have several benefits in human health. These are to
reduce the risk of cardiovascular disease and some neurological illness
including Alzheimer’s and Parkinson’s diseases (Kerdiles et al., 2017).
Moreover, DHA has substantial role in the normal operation of brain
(Horrocks and Yeo, 1999). Although, the DHA is synthetised from ALA,
unfortunately, this conversion is very limited in the human body (Patel
et al., 2020); it must be contained in the diet as essential fatty acid.
Unhealthy diets are associated with higher consumption of n-6 and
lower consumption of n-3 fatty acids. A balanced n-6/n-3 PUFAs ratio is
essential for human health. Increasing ratio of n-6/n-3 PUFAs connects
with higher risk of coronary heart disease and neurodegenerative dis­
orders (Trebaticka et al., 2020) and led to obesity (Simopoulos et al.,
2016). Currently, the n-6/n-3 PUFAs ratio in the human diet has
changed to 15–20:1 or even higher (Patel et al., 2022). Recently, the
recommended ratio of the n-6/n-3 PUFAs is less than 4 (Simopoulos
6
Small Ruminant Research 226 (2023) 107058
et al., 2016). In the present study, all cheese samples’ n-6/n-3 PUFAs
ratio were lower than the recommended ratio for human nutrition,
whereas this ratio was favourable in MA group.
The ripening only slightly affected the fatty acid profile of cheese in
present study. Earlier studies on other type of cheeses also found no
significant effect of ripening on the fatty acid profile (Gnädig et al.,
2004; Bonanno et al., 2013). In addition, during ripening, the rumenic
acid concentrations were slightly changed from 0.31 g/100 g of FA at
2nd day to 0.32 g/100 g of FA at 4 weeks, then significantly (P < 0.05)
increased to 0.34 g/100 g of FA at 8th weeks compared with beginning
of ripening, while the DHA concentrations were stable during ripening.
The concentrations of odd- and branched-chain fatty acids (OBCFAs)
markedly decreased in the cheese of the MA goats fed daily 10 g/head
marine algae supplementation compared with animals of C group. The
OBCFAs mostly originated from the rumen bacterial populations (Har­
foot, 1981). These fatty acids are associated with the health indicator of
bacterial populations in rumen. The OBCFAs absorbed in the gut
throughout the intestinal wall and get into blood flow, then uptake of
blood circulating FA by the mammary gland led to the appearance of
OBCFAs in milk fat. Most abundant odd-chained fatty acids are the
pentadecanoic (C15:0) and heptadecanoic (C17:0) acids, in agreement
with other reports (Vlaeminck et al., 2006; Nudda et al., 2021), that are
mostly derived from ruminal bacteria cell wall (Nudda et al., 2021).
However, several epidemiological studies suggest that the OBCFAs
(mainly C15:0 and C17:0) are associated with reduced disease risks of
CHD, hepatic steatosis and type 2 diabetes (Jenkins et al., 2015; Ima­
mura et al., 2018). The marine algae inclusion contains high concen­
trations of LC-PUFA, which are inhibit the step of hydrogenation from
trans C18:1 to 18:0 causing higher levels of trans C18:1 fatty acid and
other specific biohydrogenation intermediates, which have negative
effect on the abundance of bacterial population in rumen (Toral et al.,
2017; Białek et al., 2018).
Moreover, Nudda et al. (2021) reported that milk processing caused
significant difference between the OBCFAs concentrations of milk and
cheese or other dairy products. Nudda et al. (2021) also stated that
different microbial cultures which are interesting in the dairy industry,
may change the raw milk OBCFAs concentrations in cheese or in other
dairy products after processing. Although, the background of this
change is still unknown. In addition, the odd- and branched-chain fatty
acids concentrations remained constant in the cheese during ripening in
the present study.
The total solids content in VP group (50.6 g/100 g cheese) was lower
than UP cheeses in present study. This is most likely due to the fact that
the cheeses were packaged and ripened in low moisture permeability
plastic bags. This is in concordance with the previous results of
(Agboola, Radovanovic-Tesic., 2002), Frau et al. (2021) and Darnay
et al., (2019). The contents of fat and fat in TS were similar to TS con­
tent. In contrast, the protein content was stable during the maturation;
this probably reflected to the low level of proteolysis during maturation.
Earlier, Barać et al., (2021) found less intensive proteolysis in vacuum
packaged cheese than in cheese ripened in brine. In addition, vacuum
packaging prevents moisture loss, which may have economic benefit for
cheese makers, when cheeses are sold by their weight.
Finally, in present study, the levels of short chain fatty acids (C4:0 -
C12:0) were lower in VP cheeses; parallel to this, the concentrations of
medium and long chain fatty acids were higher compared with UP
cheeses. These results suggest an indication of the lower intensive
lipolytic changes in VP cheeses than unpackaged cheeses during
ripening. It was concordance with results of Barać et al., (2021). During
lipolysis, the triglycerides in cheeses hydrolysed by the milk-originated
lipases, which resulted in higher concentrations of fatty acids in cheese
during ripening (Sausa et al., 2001). The lipolytic activity was influ­
enced by other factors such as the starter microorganisms (Mallatou
et al. 2003). Moreover, the low concentration of short chain fatty acids
may influence the cheese flavours. The flavour of cheese is mainly
produced during the maturation of cheese from proteolysis (amino acids
Á. Bodnár et al.
and peptides) and from lipolysis (mainly short and medium-chain fatty
acids) (Fox, 1989; Molimard et al., 1997). The occurrence of short-chain
fatty acids in cheese contributes to its sensory qualities due to their
lower thresholds of perception compared to long-chain fatty acids
(Laskaridis et al., 2013). However, Efthymiou and Mattick (1964) re­
ported that objectionable rancid flavour developed in cheeses with high
levels of C12:0 and other longer fatty acids.
5. Conclusions
Dietary supplementation of 10 g/head/d marine algae had no effect
on the milk composition, however, feeding marine algae significantly
influenced the cheese composition without affecting the fat and the fat
in total solids contents. As expected, marine algae supplementation
improved the concentrations of DHA and rumenic fatty acids, further­
more, the ripening elevated the concentration of rumenic acid in cheese.
Moreover, marine algae inclusion in diet had strong effect on concen­
trations of odd- and branched-chain fatty acids, with significantly
decreasing their concentrations. In addition, the results clearly demon­
strate that the vacuum packaging has a notable impact not only on the
cheese composition, but also on the fatty acids profile of the cheeses.
Author contributions
ÁB and FF: Conceptualization, Methodology. ÁB and IE: Data
collection. ÁB and FP: Data curation, Data analysis, Data Visualization.
ÁB, IE, JK, and FP: original draught preparation. All authors: Writing –
review & editing. All authors agreed with the pre-published version of
this paper.
Declaration of Competing Interest
The authors declare no conflict of interest.
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