Vol.4, No.5B, 78-83 (2013) Agricultural Sciences
Survival of encapsulated probiotics through spray
drying and non-refrigerated storage for animal feeds
Norfahana Abd-Talib1,2, Siti Hamidah Mohd-Setapar2*, Aidee Kamal Khamis1, Lee Nian-Yian2,
Ramlan Aziz1
1Institute Bio Product Development, Universiti Teknologi Malaysia, UTM Johor Bahru, Johor, Malaysia
2Centre of Lipid Engineering and Applied Research (CLEAR), Universiti Teknologi Malaysia, UTM Johor Bahru, Johor, Malaysia;
*Corresponding Author: sitihamidah@cheme.utm.my
Received 2013
The objective of this study is to verify potential of
various types of microorganisms during spray
drying and non-refrigerated storage that can be
enhanced substantially by selecting suitable pro-
tective colloids. Four selected probiotics tested
are Lactbacillus plantarum B13 and B18, which
are the bacteria probiotics and Kluyveromyces
lactis and Saccharomyces blouradii, non-bac-
teria probiotics. Two levels of experiment occur
starting with formulation study of encapsulation
agent followed by the viability study of different
probiotics after spray dry and two weeks nonre-
frigerated storage. The formulation of 30% of
gum Arabic, 15% of gelatin and 45% of coconut
oil can homogenize well at least for two hours
and can produce acceptable dried product (be-
low 4% of moisture content) at low outlet tem-
perature (70 - 75). K. lactis, S. blouradii
gives 2.57% and 2.4% of viability percentage
after spray drying process and 25.84% and
2.04% after two weeks nonrefrigerated storage
respectively. The colonies of non-probiotics
bacteria after both conditions are between 1010
and 106 cfu/mL which is among the accepted
level for industrial application. However, the
survival of probiotics in a spray-dried form dur-
ing non-refrigerated storage is higher at low of
moisture content compared to others.
Keywords: Probiotics; Survival Rate; Spray Drying;
Nonrefrigerated Storage
Probiotics have been recently defined as “live microbes
which transit to the gastro-intestinal tract and having posi-
tive effect s to th e health for the consu mers [1]. I t have b een
applied in aquaculture for controlling diseases,
enhancing the immune response, supplementing or even
in some cases replacing the use of anti microbial. The
use of probiotics as farm animal feed supplements dates
back to the 1970' and originally incorporated into feed to
increase the animal's growth and to improve its health by
increasing its resistance to disease. It was assumed that
the effectiveness of probiotics were related to the
gastroin-testinal tract and effects on incidence of diarrhea
and other gut infection s. According to literatu res show in
Table 1, there are some probiotic strains which can give
positive effect to various typ es of animals.
Some of other probiotics which used in animal feed
are microscopic fungi such as strain of yeasts. There are
billion strains of yeasts that are supposed to be explored
in animal feed study. The strains like Saccharomyces
boulardii and Kluveromyces lactis are having high po-
tential as a probiotic that can improve animal health.
Yeast seemed to facilitate increased mobilization of body
reserves and to increase milk fatty acid production of
ruminants. Therefore, it was possible to apply yeast’s
strain as probiotics strain in powder form of animal feed
in line with the effectiveness in animal health. For exam-
ple camel calves can improve weight gain, average daily
gain and feed utilizations after apply yeast as supple-
mentation diet in different forms [9].
It is known that probiotics were very useful for animal
but the preparation of animal feed that containing probi-
otic especially in powder form are not easy. To ensure
the amount of probiotics cu lture required can achiev e the
target; some of the factors are to be tested first in order to
increase stability or viability of the probiotic products.
Encapsulation agent is the formulation of single or com-
bination of wall material that can resist the condition
along the journey. Encapsulation is a process in which
tiny droplets or particles are wrapped with a protective
Copyright © 2013 SciRes. Openly accessible at http://www.scirp.org/journal/as/
N. Abd-Ta lib et al. / Agricultural Sciences 4 (2013) 78-83 79
coating yielding capsules for countless application. Sev-
eral methods of encapsulation of probiotic have been
reported by [10]. The objective of the research is to dem-
onstrate the various types of probiotics microorganisms
during spray drying and storage at room temperature by
selecting suitable protective colloids as an encapsulation
2.1. Stability of the Feed Composition
The stage one of this experiments was conducted by
the conventional try and error method. The several feed
formulations were choose which with high advantages in
spray dryer process and animal feed industry. At the first
stage, the homogenize mixture without probiotic cell was
hold for 2 hours to test stability of the mixture by using
homogenizer (Heidolth, DIAX 900, Germany). This is
holding time of the feed formulation in the feed bottle
during spray dry (Pilot Spray Drying Plant PSD-00,
Hemray Enterprise, Bombay). The homogenized mix-
tures were proceed for drying process in the spray dryer
at inlet temperature 110℃ and outlet temperature of air
70- 75℃ and should give the moisture content below
4%. Details of the various wall materials used are shown
in Table 2.
Table 1. Some probitiocs strain with the benefits to animals.
Animal Probiotics Functions References
Lactobacilli spp.
Lactobacillus sakei) Enhance the host protection
against pathogens. [2]
reuteri (33%)
crispatus (18.7%)
salivarius (12.3%)
To maintain balanced
microbiota and to reduce
potential pathogens [3]
Pig Enterococcus
Great production of specific
antibodies against
salmonella enteric [4]
acidphilus and
Reducing population of
Clostridium perfringers
which can lead to necrotic
enteritis prevention
To promote growth a n d
enhance immunity and
resistance against
Streptococcus sp. and an
Piglets Lactobacillus
Improve gro wt h
performance without
affecting the
gastrointestinal ecology.
acidophilus and
Body weight increased [8]
Table 2. Formulation of emulsion.
A 30% gum Arabic, 15% gelatin, 45% coconut oil
B 30% gum Arabic, 15% gelatin, 45% lecithin
C 30% gum Arabic, 15% lecithin, 45% coconut oil
D 15% gelatin, 30% lecithin, 45% coconut oil
E 15% gelatin, 30% maltodextrin, 45% coconut oil
F 30% maltodextrin, 15% le ci th in , 45% coconut oil
G 15% gelatin, 30% maltodextrin, 45% lecithin
H 15% gum Arabic, 45% gelatin, 7.5% lecithin,
67.5% coconut oil
I 45% gum Arabic, 15% gelatin, 7.5% lecithin,
67.5% coconut oil
2.2. Survival of Different Types of Probiotics
During Spray Dry
The second stage of stud y was focused on the viability
of different type of probiotics on the spray dry condition
and storage at 25℃ in two weeks. Four different types
of probiotics microorganisms were chosen to investigate
the viability, stability with wall material used, condition
of spray dry, and two weeks non-refrigerated condition.
The chosen probiotics were Lactobacillus plantarum B13
and B18, Kluyveromyces lactis and Saccharomyces
blouradii. The mixtures of wall material s were mi xed wi t h
deionised water and autoclave a 121℃ for 15 minutes
(water phase). The coconut oil was sterilized filter and
then the probiotic culture was dispersed into coconut oil
using ho- mogenizer at the lowest speed (3000 rpm) for
10 - 15 minutes (Zen tr ifugen , Hettich,D-78532,Tuttlinge
n, Ger- many). Then, the coconut oil were mixed up with
water phase mixture by using homogenizer at 3000 rpm
for 10 minutes in sterilized condition.
2.3. Viability Analysis
1 mL of each formulation was transfer into a universal
bottle and 9 mL sterilized water is added. The solution is
serially diluted until dilution factor of 1010. 0.01 mL is
taken from all the dilution factors and transferred into the
petri dish contain medium agar for each type of probiotic
respectively. After that, petri dish is incubate for 24 hours
at 37℃ for Lactobacillus plantarum and Kluyveromyces
lactis and 30℃for 24 hours for Saccharomyces bloura-
dii. After 24 hours, the number of colony forming is cal-
culated with naked eyes. The moisture content of spray
dried powders was determined by oven drying at 102℃
until constant weight was obtained.
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N. Abd-Ta lib et al. / Agricultural Sciences 4 (2013) 78-83
3.1. Formulations study of Encapsulation
Try and error method carried out to get the suitable
emulsion formulation in order to get the good product.
Based on previous studies, four basic materials which
can contribute on production of good powder product
were selected. After doing some try and error formula-
tion and stability checked, the observ ation were shown in
Table 3.
After doing the observation, the stability test resulted
that Formulation A and Formulation D show a highest
potential in maintaining the stable characteristics after
keeping for two hours. The maximum duration of spray
drying is about two hours. So, at least the formulation
can homogenized well at the duration time is more than
enough. Lecithin was an economic an effective wall ma-
terial but unfortunately it cannot homogenize well with
other wall material selected. The ability of solid per-
centage used in the solution has been checking up after
selection of the suitable emulsion formulation successful.
Formulation A and Formulation D were the only two
formulations that have been selected to the next part of
Table 3. Stability of different of encapsulation agent formula-
Formulations Ingredients Stability Observations
A 30% gum Arabic,
15% gelatin,
45% coconut oil
After homogenize, milky whit e
and stable solution produced
B 30% gum Arabic,
15% gelatin,
45% lecithin
After homogenize, 70 percent
from solution precipitated.
C 30% gum Arabic,
15% lecithin,
45% coconut oil
After homogenize, 50 percent
from solution precipitated.
D 15% gelatin,
30% lecithin,
45% coconut oil
After homogenize, m ilk y y ell ow
and stable solution produced
E 15% gelatin,
30% maltodextrin,
45% coconut oil
After homogenize, milky whit e
and unstable solution produced
F 30% maltodextrin,
15% lecithin,
45% coconut oil
After homogenize, 70 percent
from solution precipitated
G 15% gelatin,
30% maltodextrin,
45% lecithin
After homogenize, 50 percent
from solution precipitated.
15% gum Arabic,
45% gelatin,
7.5% lecithin,
67.5% coconut oil
After homogenize, 30 percent
from solution precipitated.
45% gum Arabic,
15% gelatin,
7.5% lecithin,
67.5% coconut oil
2 layer of milky a n d p l a i n
solution produced
3.2. The Effect of Feed Formulation on
Moisture Content
Formulations A and D were the only two emulsions
that can be homogenize well in the long time given. After
spray drying process, Formulation A give more positive
result than Formulation D but still cannot achieve the
target. According to Desmond et al.,[11], the level of
moisture content required for prolonged powder storage
life and stability are at least four percent. Different of
solid percentage selected and being tested to impro ve the
dried dairy product and the parameters used also can
contribute to the improvement of dried product quality.
By increasing the solid percentage used in the solu tion,
it shows the positive result for formulation A. The mois-
ture content decreased to a very stable value. Because of
the suitable outlet temperature have been proven by
Meng et al.,[12] which the best value to dry the probiotic
culture optimally is around 70 - 75. Inlet tempera-
ture have been controlled based on the outlet temperature
selected. The inlet temperature was maintained at 110
in order to obtain an outlet air temperature in the range of
70 - 75. Therefore, the only parameters that can be
manipulated here is feed flowrate. Based on previous
part of study, wet product produced if using 120 rpm of
feed rate. It preferred to decrease the feed rate to produce
the better form of powder. By using 100 rpm of feed rate,
the product form gives a very low moisture content per-
centage that was below four percent.
For formulation D, when the amount of the solid par-
ticle in the formulation increases, the moisture content
was increased up to 15%. The high stickiness of the
powder on the walls of drying chamber was observed.
The results were unreliable and the formulation D dis-
qualified to the n ext part of study automatically.
3.3. Spray Drying of Different Types of
Probiotic Microorganisms
Different types of microorganisms selected based on
the priority in animal health enhancement. Other re-
searcher has been proved that Lactobacillus strain and
yeast strain can improve animal health. Encapsulations
of the entire microorganisms by the best-selected wall
material and the best-feed flowrate from the previous
part of study have been done. The objective of this part
of the study is to check the viability of different probiot-
ics such as L. plantarum strain 13 and 18, K.lactis and
Figure 1 shows the viability of the different types of
microorganisms through spray dryer condition and two
weeks of non-refrigerated storage at 25. It directly
shows the yeast strain of Kluyveromyces lactis give
highest survival rate on both situation. The recom-
mended minimum numbers of viable microorganisms in
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N. Abd-Ta lib et al. / Agricultural Sciences 4 (2013) 78-83
Copyright © 2013 SciRes. http://www.scirp.org/journal/as/
probiotic food for efficacy are 106 cfu/g [13]. The reten-
tion of high viability during drying and storage presents
particular challenged and can be classified as a major
problem in commercial probiotic prod uction [12]. In this
research, the inlet, outlet temperature and feed rate was
constant at 110, 75 and 100 rpm respectively. Gen-
erally, it was found that percentage of survival of Lacto-
bacillus plantarum B18 strain was higher than others
during spray dry with the similar carrier. However, it
gives lowest viab ility dur ing storag e. It was believed that
better survival of Lactobacillus plantarum B18 may be
attributed to less sensitivity of this organism when ex-
posed to heat. Only Kluyveromyces lactis can gives a sta-
ble survival characteristic than others.
Openly accessible at
Figure 1 proved the low potential of Lactobacillus
plantarum B13 strain through spray dry and two weeks
storage. The number of cells reduced from 1.28 × 108
cfu/ ml to 2.1 × 106 cfu/ml and final ly to 3.0 × 105 cfu/ml.
This type of bacteria not effective and not suitable for
comercial used with the wall material of gum Arabic, gela-
tin an d coconut oil. The resu lt will be better with the other
mi xture as an e ncapsulation agent. The resistance was very
low even only in the spray dry condition. It was not appli-
cable to select this type of bacteria to improve the animal
feed if the carrier cannot match well with the bacteria.
Lactobacillus plantarum B18 resulted the best viabil-
ity after spray drying and storage at room temperature
with combination of gum Arabic, gelatin and co conut oil
as a wall material. Therefore, it was only reliable in spray
dry condition not during storage. The figure also shows
how the cells drastically decrease just after spray dry and
after two weeks of storage compared to before. The
number of cells reduced from 3.25 × 107 cfu/ml to 2.15 ×
107 cfu/ml and after two weeks storage, the total de-
creased to 4 ×102 cfu/mL. The viability of the bacteria
along the storag e depends on the sensibility of the b acte-
ria to oxygen and the ability of carrier material as a pro-
tector. This is because of the low moisture content of the
powder about to affect the viability of the bacteria of
Lactobacillus Plantarum B18 to be decreased drastically.
The survival of Kluyveromyces lactis comparing with
the other bacteria, it gives the most reliable where the
rate of survival of the cell during spray dry and even
storage were higher. Number of cells reduced from 2.99
× 109 cfu/ml to 7.4 × 107 cfu/ml and 1.9 × 107 cfu/mL
after two weeks storage at 25. The ability of this cell
with the carrier which consists of gum Arabic, gelatin
and coconut oil can be commercialized. Different from
human, a probiotic dose between the ranges of 106 to 107
cfu/g in the animal feeds is more than enough [13]. Be-
cause of that, the value resulted for Kluyveromyces lactis
shows the ability to improve the probiotic in the animal
feeds. The other strain of yeast name Saccharomyces
blouradii also gives positive result for this study. The
Figure 1 proved the amount of cell culture that can
maintained in heat and storage. The value of cell culture
can give good impact on animal feed industry which
were reduced from 2.1 × 1010 cfu/ml to 3.9 × 108 cfu/ml
and after two weeks of storage at 25, it becomes 7.82
× 106 cfu/ml.
no. of bacteria (cfu/mL )
type of bacteria
before spray
after spray
after 2 weeks storage
Figure 1. The colony of different probiotics after spray dry and two weeks storage.
N. Abd-Ta lib et al. / Agricultural Sciences 4 (2013) 78-83
mo istu r e c o n te n t ( %)
type of bacteria
Figure 2. Moisture percentage of powder form of different probiotics after drying using spray dry.
3.4. Moisture Content of Different
Spray-Dried Probiotics
The carrier used during spray drying of probiotics is
known to have an influence on storage ability. Because
of this study deal with the different types of probiotics,
the ability of the encapsu lation agent to protect probiotic
microorganisms during drying is important characteristic.
Figure 2 had shown the graph of moisture content per-
centage in the different type of dried form probiotics.
Moisture content values give the big impact to viability
of probiotic microorganisms during storage. Spraydried
cultures retain viability for longer at low temperature;
however, refrigeration is expensive to supplier and re-
tailer of product. There is a need to produce the probiotic
bacteria cultures that are stable in ambient temperature
This study demonstrated the possibility of producing
dry probiotic that suitable as an animal feed using spray
drying. Using combination of gum Arabic, gelatin and
coconut oil with inlet and outlet temperature, feed rate
and solid percentage in the solution is 110, 75, 100
rpm and 13.5% respectively give the best powder form
of product which is below 4%. A bacterial survival rate
during both spray dry and two weeks non-refrigerated
storage condition have a wide range of percentages.
Kluyveromyces lactis and Saccharomyces blouradii gives
the big potential result for this study, wh ich is suitable to
use in animal feed industry. Survival of probiotic in a
spray-dried form during storage is higher at lower mois-
ture content.
The Universiti Teknologi Malaysia is thankful for the grant given to
Dr. S.H. Mohd-Setapar’s project (Q.JI3000.7125.00H02) under Re-
search University Grant Scheme, part of which enabled this review
article to be prepared. We are grateful for the contribution of all people
involved in this paper pr ep aration.
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