The application of simple technology

for making yogurt powder to improve

biochemical blood profile of broiler
Cite as: AIP Conference Proceedings 2219, 070009 (2020); https://doi.org/10.1063/5.0003199
Published Online: 05 May 2020

Lovita Adriani, I. Made Joni, Chitra Kumalasari, and Ronny Lesmana

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AIP Conference Proceedings 2219, 070009 (2020); https://doi.org/10.1063/5.0003199 2219, 070009

© 2020 Author(s).
 The Application of Simple Technology for Making Yogurt
Powder to Improve Biochemical Blood Profile of Broiler
Lovita Adriani1, a), I Made Joni2, Chitra Kumalasari1 and Ronny Lesmana3, 4
1
Departement of Animal Nutrition, Faculty of Animal Husbandry, Universitas Padjadjaran,
2
Departement of Physics, Faculty of Manthematic and Natural Science, Universitas Padjadjaran,
3
Division Physiology, Departement of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran,
4
Biological Activity, Molecular Physiology Laboratory, Central Laboratory, Universitas Padjadjaran,
Jl. Raya Bandung Sumedang KM 21, Jatinangor, West Java 45363, Indonesia.

a)
Corresponding authors: lovita_yoghurt@yahoo.co.id

Abstract. Probiotics have several positive benefits, such as reducing pathogenic bacteria population, providing enzymes
which help for food digestion, and lowering blood lipid levels. Best formulation can be formulated by combining powder
with a mixture of L. bulgaricus: L. acidophilus: S. thermophilus = 1: 1: 1. Yogurt formulation using a simple coated
technology with 5% of maltodextrin. Aim of our study, we measured the effectiveness of formulated yoghurt powder coated
with maltodextrin to control broiler blood lipid profile. We developed soy-yogurt using probiotic mentioned and utilizing
the blower to accelerate the drying process.which will facilitate feeding process at livestock by mixing with feed rations
and extending the storage time. Total bacterial plate count after powder processing showed that there was a decrease of
the number of Lactic Acid Bacteria for each treatment, ranging from T-1 4.21 x 103; T-2 3.49 x 106; T-3 3.85 x 103, and T-
4 7.09 x 102 from total bacteria 1.55-6.75 x 109. Then, sample are divided into four treatments that were T-1 =Basal Diet,
T-2 = Basal Diet + 2% Probiotic ( 100% Fermented- Cow Milk), T-3 = Basal Diet + 2% Probiotic ( 50% Fermented-Cow
Milk + 25% Fermented-Soy Milk), T-4 = Basal Diet + 2% Probiotic ( 50% Fermented-Cow Milk + 25% Fermented-Soy
Milk + 25% Fermented-Mung Bean).The result showed that there was a significant result on blood cholesterol while the
other lipid treatments had no significant difference.

INTRODUCTION

Probiotics is microorganism that can modify the microflora in the body, it can replace the pathogenic microbe with
non-pathogenic microbe for the body in the human’s intestine [1]. Probiotics are microorganism consumed either in
the form cell or fermented product gives the advantage for improving indigenous microflora character [2]. Besides on,
probiotics are microorganism has the ability to improve the ecosystem of intestinal flora and to maintain the host.
Some probiotics have to reach colon in order to decrease the number of pathogenic bacteria in the colon also shows
its physiology role in maintaining the balance microflora at the gastrointestinal tract [3]. Efforts to reduce cholesterol
and triglyceride levels in livestock, especially in broiler chickens need attention, one of which is by providing
probiotics.
The provision of probiotics can use some methods. Probiotic supplementation using fermentation of milk, soy
milk, and green-bean milk gives the high results on broiler’s blood biochemistry. The administration of probiotic to
livestock experience difficulty in feeding because of that product in the form of liquid. Therefore, it needs technology
to change the product to be powder. One of its methods is the application of simple drying technology by using an
oven with a thermometer and blower. The objective of powder with a mixture of L. bulgaricus: L. acidophilus: S.
Thermophilus: Bifidobacterium = 1: 1: 1: 1. The making yogurt used a simple technology where yogurt was coated
by 5% of maltodextrin. This is carried out to supply nutrient as media for microbe. The purpose of the drying process
was to facilitate in feeding process at livestock because it could be mixed with feed ration so that it extended the
storage time.

2nd International Conference and Exhibition on Powder Technology (ICePTi) 2019

AIP Conf. Proc. 2219, 070009-1–070009-5; https://doi.org/10.1063/5.0003199
Published by AIP Publishing. 978-0-7354-1975-9/$30.00

070009-1
 Probiotics have many functions for the body, especially for improving the ecology bacteria at intestine , preventing
intestinal cancer and increasing the digestibility of nutrients [3-4]. Besides that probiotics have the ability to increase
the activity of digestive enzymes in the intestine so that the absorption of nutrients will become maximum. It can
increase the area for absorption nutrient at intestine with influence the intestinal anatomy like density and size of small
intestine villi [5]. In this research, milk either cow’s milk or soy is fermented by probiotic bacteria. Probiotics are
fermented with several materials like soy milk, cow’s milk then it will be searched its the best combination. Soy is a
group of oligosaccharides contains sucrose and raffinose it is difficult to digest and to absorb by the intestine.
However, it helps a growth substrate for non-pathogenic bacteria in the intestine [6]. Lactic acid bacteria in fermented
soy milk have the ability in increasing isoflavone digestibility [7]. Isoflavones are secondary metabolite compounds
that are usually synthesized by plants. In the soybean, there are 2 to 4 mg/g isoflavone contents. Kind of isoflavones
are genistein, daidzein and glisten [8]. Antioxidants have been known to have the ability in improving intestinal villi
because it gives an impact on increasing absorption area. Fermentation of lactic acid bacteria also results in hydrolysis
of soy protein into short peptides like Phe-Asp-His-Val-Glu and PheAsn- His-Leu-Asp-His, which are able to decrease
DPPH free radicals, thus acting as antioxidants [8].
Taken together, probiotic supplementation based on milk -coated powder has not been done will manage the blood
lipid profile in broiler. Therefore, it needs to be done the research to obtain information about the number of bacteria
in probiotic.

MATERIALS AND METHODS

Birds, Housing, and Diet

A total of 100 day-old broiler chicks were used in the experiment. The experiment was conducted for 28 days.
The fermented milk as a probiotic was mixed in rations. The composition was based on the type of fermented milk of
each treatment. The dose of fermented milk is 2% of the total rations. Cow and soybean milk was used as the basic
ingredients to be fermented. The fermentation process used bacteria, namely Streptococcus thermophilus,
Lactobacillus bulgaricus, Lactobacillus acidophilus and Bifidobacteria bifidum. The experiment design used
Completely Randomized Design with 4 treatments and 5 replications. The total experimental units were 20 and each
unit contained 5 day-old chicks. The treatments consisted of (T-1) basal ration (without fermented milk), (T-2) basal
ration mixed with the cow milk at a dose of 2% of total ration, (T-3) basal ration mixed with the fermented cow milk
and fermented soybean milk (ratio 1:1) at a dose of 2% of total ration, (T-4) basal ration mixed with the fermented
cow milk, soybean milk, and mung bean milk (ratio 2:1:1) at a dose of 2% of total ration.

Processing of Probiotic Powder

Fermentation of cow milk, soy milk, mung bean milk were done in the physiology research laboratory, Animal
Husbandry Faculty , Universitas Padjadjaran. Fermentation milk was performed using Streptococcus thermophilus,
Lactobacillus bulgaricus, Lactobacillus acidophilus, and Bifidobacterium bifidum. Fermented milk is mixed with
maltodextrin DE 10-12 5% as a nutrient for microorganism to stay alive. Drying probiotic used the dry oven (simple
method) with a temperature of 40oC for 25 hours. The probiotic powder is given to broilers in ration correspond with
treatments at a dose of 1.25% of the total ration [9].

Blood Parameters

Blood samples (3ml) were collected from 20 broilers per replicate, then were analyzed for cholesterol and
triglyceride levels at the Physiology and Biochemistry Laboratory, Animal Husbandry Faculty, Universitas
Padjadjaran. CHOD-PAP (Cholesterol Oxidase Phenylperoxidase Amino Phenozonphenol) method was used to
analyze the cholesterol. GPO (Glycerol-3-Phosphate Oxidase) method was used to analyze the triglycerides.

070009-2
 Statistical Analysis
Data were tabulated and analyzed using SPSSS ver 13.00 with one-way analysis of variance (ANOVA). The
significant differences among different treatment means were investigated using Tukeyʼs test by considering
differences significant at p<0.05.

RESULT AND DISCUSSION

Table 1 is shown the data about cholesterol and Triglyceride :
TABLE 1. The effect of various probiotic fermentation media on biochemical blood broiler
No. Variable Treatment P. Value
T-1 T-2 T-3 T-4
b b b
1 Cholesterol 155.08 151.68 140.28 119.38a <0.001
2 Triglyceride 155.075 151.69 140.283 119.37 >0.05
Note : T-1 (Basal Diet)
T-2 (Basal diet +2 % Probiotic (100% Fermented-cow milk)
T-3 (Basadiet + 2% Probiotic (50% Fermented-cow milk + 50% Fermented-soy bean milk)
T-4 (Basal diet + 2% Probiotic (50% Fermented-cow milk + 25% Fermented soy bean milk + 25% Fermented Green
Bean Milk)

We observed that all treatment gives a significant result on cholesterol, and the best outcome come from treatment
is 119.38 in T-4. We did not observe any significant alteration on triglyceride levels, however, there is a trend showed
T-3 give a positive impact that is T-3 stimulate the lowest value of triglyceride levels (Table1).

180
155.08 155.075 151.68 151.69
160
140.28 140.283
140
119.38 119.37
120
mg/dL

100

80

60

40

20

0
T-1 T-2 T-3 T-4

Cholesterol Triglyceride

FIGURE 1. Illustration of data in Table 1 regarding the effect of treatment on cholesterol and triglyceride

Cholesterol

The results of statistical analysis of blood cholesterol levels ranging from the highest to the lowest T-1 = 155.08
mg/dl, T-2 = 151.68 mg/dl, T-3 = 140.28 mg/dl, and T-4 = 119.38 mg/dl. Statistical analysis showed that the results
were significantly different (P <0.05) to decrease blood cholesterol levels. T-2 and T-3 treatment had no significant

070009-3
effect, but T-4 significantly affected the treatment compared to T1 in reducing blood cholesterol levels (Figure 1).
This finding might indicate that probiotics are capable of producing cholesterol reductase enzymes. The cholesterol
reductase enzyme can convert cholesterol to coprostanol, a type of sterol that cannot be absorbed by the intestine.
Coprostanol is a natural steroid that can be produced by bacteria in the lower intestine of humans or animals and
released through feces [10] .
The treatment of T4 (50% of fermented cow's milk, soybean milk 25%, and mung bean milk 25%) reduced broiler
chicken blood cholesterol levels. The difference in basic materials will affect microbial activity in culture because the
basic ingredients affect the growth of lactic acid formation [11]. This decrease occurs because in P3 using mung beans
(Vigna radiata L) that contained flavonoids, phenolic acids, proteins, amino acids, carbohydrate fractions, lipids,
minerals, and other metabolites [12]. Those can act as antioxidant, antitumor, anti-inflammatory, hepatoprotective,
antimicrobial, diuretic, hypoglycemic, and able to reduce cholesterol levels [13].
Soybean nutrition composition per 100 g of seeds including 3.7 g protein, 2.8 g carbohydrates, 0.6 g fiber, 120 mg
calcium, 0.2 mg vitamin B12, and 2.2 g total fat. There are also 0.4 saturated fatty acids, 0.5 g monounsaturated fatty
acids, 1.3 g polyunsaturated fatty acids consisting of linolenic acid (omega 6) 1.2 g and α-linolenic acid (omega 3) 0,
2 g (Burssens et. al., 2011).Flavonoid-compounds in soy milk and mung bean reduced cholesterol levels in blood.
Flavonoid is one of the phytochemical groups that have the same structure, namely polyphenols, whose mechanism
can reduce cholesterol levels due to HMG-CoA (Hydroxy Methyl Glutatyil-CoA) reductase activity, reduce the
activity of the enzyme acyl-CoA cholesterol acyltransferase (ACAT), and reduce cholesterol absorption in the
digestive tract [14]. Isoflavone compounds in soy milk and mung bean are also reported to reduce cholesterol levels
in addition to flavonoids. Isoflavones are included in the class of flavonoids which are polyphenolic compounds in
soy milk. The mechanism for reducing cholesterol by isoflavones is explained by the effect of increasing fat cell
catabolism for energy formation which results in a decrease in cholesterol content [15]. The conclusion shows that
blood cholesterol levels decrease and are significant. This finding occurs in T-4 treatments, which is 50% fermented-
cow milk, 25% soy milk, and 25% mung bean.
The study had shown that the probiotics with media of fermented-cow milk or a mixture of cow milk, mung bean,
and soy milk decreased of blood cholesterol and only T-4 that has shown a significant result.

Blood Triglyceride

The average blood triglyceride level of broiler chickens <150 mg / dL [16], Other previous studies have reported
that normal averages of triglyceride levels were 47.2-162 mg/dL. This shows that blood triglyceride levels in each
treatment are in the normal range.
The decrease of blood triglyceride levels in broiler chicken is due to the supplementation of 100% fermented cow's
milk (T-2), (T-3), and (T-4) in the diet due to the ability of probiotic bacteria to produce statins that play a role in the
biosynthesis of triglycerides. In line with research done by Cavallini et al. [17], probiotics can produce statins, namely
3-hydroxy-3-methyl-glutaryl-CoA reductase inhibitors (HMG-CoA reductase) which are cholesterol biosynthesis
regulating enzymes, lowering LDL, VLDL, and blood triglyceride levels. The mechanism of decreasing triglyceride
levels by statins begins when the inhibitor reduces cholesterol concentration in hepatocytes and increases the
performance of LDL-receptors that are closely related to VLDL components so that triglycerides will be reduced [18].
The occurrence of inhibition of triglyceride synthesis in the liver and small intestine will result in a decrease in blood
triglyceride levels. The reduction of fatty acid synthesis in the liver is the main factor causing the decline in
triglyceride synthesis in the liver which results in a reduction of the concentration of triglycerides in plasma [19].
The prior research proves that supplementation of 75% fermented cow's milk + 25% fermented soy milk decreased
triglyceride in plasma due to the closely related decrease in the number of pathogenic bacteria in the intestine. Probiotic
bacteria such as Lactobacillus acidophilus and Bifidobacterium can inhibit the growth of pathogenic bacteria [20].
Moreover, probiotic administration created a balance of intestinal microflora because of the presence of BAL in the
intestine which creates an acidic atmosphere that suppresses the growth of pathogenic bacteria in the small intestine
[21]. The acidic environment inhibits the secretion of the lipase enzyme, therefore the synthesis of fatty acids in the
digestive tract decreases and causes fat to be brought to the liver to decrease which results in a reduction in blood
triglyceride levels. Finally, it has been shown that simple drying technology can decrease the number of microbes,
but it may improve that the content of other substances such as flavonoids, antioxidants, vitamins, and fatty acids
which may play significantly in biological function.

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 CONCLUSIONS
Supplementation probiotics-based fermented milk with 50% fermented cow's milk , 25% fermented soybean milk
and 25% fermented green beam milk is the best result for improving blood lipid , also supplementation using mixed
milk in all treatments showed better results than control. Supplementation probiotics in powder form are recommended
because it shows better results.

ACKNOWLEDGEMENT

The authors thankful to Academic Leadership Grant, Universitas Padjadjaran for the financial support of this
research. The authors also acknowledge Norman Billi, who has helped in the experiment and analyzing data.

REFERENCES

1. Metnikoff, Probiotics : A Critical Review (Horizon Scientific Press, England, 1908).

2. R. Fuller, Probiotics 2 Application and Practical Aspects, 1st Edition (Springer Netherlands, Berlin, 1997).
3. Lovita Adriani, “Efek probiotic sebagai starter dan implikasi efeknya terhadap kualitas yoghurt ekosistem
saluran pencernaan dan biokimia darah mencit,”, Ph.D Thesis, Universitas Padjadjaran, 2006.
4. A. Savadogo, C. A. T. O. ara, I. H. N. Bassole and A. S. Traore, Afr. J. Biotechnol. 5, 678–683 (2006).
5. E. R. L. Pelicano, P. A. Souza, H. B. A. Souza, D. F. Figueiredo, M. M. Boiago, S. R. Carvalho and V. F.
Bordon, Rev. Bras. Cienc. Avic. 7,1221–1229 (2005).
6. I. Amadou, S. Olasunkanmi, Gbadamosi, Y. H. Shi, M. T. Kamara, S. Jin and G. W. L. Le, Res. J. Microbiol. 5,
372–380 (2010).
7. T. A. Larkin, L. B. Astheimer and W. E. Price, Europ. J. Clin. Nutr. 63, 238–245 (2009).
8. S. Pradana, Prospects and Benefits of Isoflavones as Phytoestrogens for Health (Jakarta, 2008).
9. L. Adriani, A. Andiany, D. Latipudin and C. Cahyani, Int. J.Poultry Sci. 6, 600–604 (2018).
10. A. N. Fadhilah, Hafsan and F. Nur, “Penurunan kadar kolesterol oleh bakteri asam laktat asal Dangke secara in
vitro,” in Seminar Nasional Mikrobiologi Kesehatan dan Lingkungan 2015, Prosiding Seminar Nasional
Mikrobiologi Kesehatan dan Lingkungan (Alauddin State Islamic University of Makassar, Makassar, 2015), pp.
174–180.
11. A. Tamime and H. C. Deeth, Yoghurt Technology and Biochemistry, J. Food Prot. 43(12), 939–977 (1980).
12. D. Tang, Y. Dong, H. L Ren and C. He, Chem. Cent. J. 8(1), 1–9 (2014).
13. J. Soucek, J. Skvor, P. Pouckova, J. Matougek, T. Slavik and J. Matousek, Neoplasma 53, 402–409 (2006).
14. J. H. Choi, M. C. Rho, S. W. Lee, J. N. Choi, K. Kim, G. Y. Song and Y. K. Kim, Arch Pharm Res. 11, 1419–
23 (2008).
15. S. Pawiroharsono, Benarkah tempe sebagai anti kanker. J. Kedokteran dan Farmasi Medika 12, 815–817 (1998).
16. H. Basmacioglu and Ergul, Turk. J. Vet. Anim. Sci. 29, 157–164 (2005).
17. D. C. U Cavallini, R. Bedani, L. Q. Bomdespacho, R. C. Vendramini and E. A. Rossi, BioMed. Cent. 8, 1–8
(2009).
18. S. M. Grundy, N. Engl. J. Med. 319, 24–32 (1988).
19. J. Scorve, A. Al-Shurbaji, D. Asiedu, I. Bjorkhem, L. Berglund and R. K. Berge. J. Lipid Res. 34, 1117–1185
(1993).
20. L. Adriani, N. Indrayati and S. Darana, CMU. J. Nat. Sci., Special Issue Agric. Natural Resour. 11(1), 383–386
(2012).
21. E. Purwati, S. Syukur and Z. Hidayat, Lactobacillus sp. Isolas dari Biovicophitomega sebagai Probiotik.
(Lembaga Ilmu Pengetahuan Indonesia, Jakarta, 2005).

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