PHYSICOCHEMICAL AND SENSORY QUALITY OF PROBIOTIC DRINK FROM SUMBAWA WILD HORSE MILK
Laksmi Nur Fajriani1), Ahmad Sulaeman2), Budi Setiawan2)
1) Postgraduate in Nutrition Science, Department of Community Nutrition, Faculty of Human Ecology, IPB University, Bogor 16680, Indonesia
2) Department of Community Nutrition, Faculty of Human Ecology, IPB University, Bogor 16680, Indonesia
*Corresponding Email: [email protected] Submitted 17 October 2023; Accepted 27 November 2023
ABSTRACT
Sumbawa wild horse milk is pure milk produced from Sumbawa horses with high nutritional content. It can survive for five months at room temperature without processing and has health benefits. However, the utilization of Sumbawa wild horse milk is still limited because of its low sensory quality due to natural fermentation, which also adversely affects its acceptability.
Adding probiotic bacteria is one of the methods to improve its quality. This study aims to develop and analyze the physicochemical and sensory quality of probiotic drink products from Sumbawa wild horse milk by adding probiotic bacteria. This study used a one-factor, Completely Randomized Design (CRD), namely different types of starters. The treatment in this study is using probiotic starter Lactobacillus rhamnosus (P1) and commercial starter Lactobacillus bulgaricus + Streptococcus thermophilus (P2). The probiotic drink from Sumbawa wild horse milk in the treatment P1 has the best results, pH 4.35, 0.55% titratable acidity, 13.10% total dissolved solids, 78.00 mPa.s viscosity, 74.66% syneresis, and 3.42x109 CFU/ml total LAB. The most preferred product based on the hedonic test is P2. P2 has a whiter color and more sour taste. The addition of probiotic starters can improve the physicochemical product, and commercial starters can improve the sensory quality of the product.
Keywords: Probiotics; sensory quality; starter; wild horse milk
INTRODUCTION
Probiotic drinks are one of the popular drinks today because they are highly nutritious and contain probiotic bacteria that can improve digestive health. Probiotics consist of live microorganisms which when administered in adequate amounts confer a health benefit on the host (FAO/WHO, 2002). The most commonly utilized probiotic bacteria are from the species of Lactobacillus sp. and Bifidobacterium (Vlasova et al., 2016). Currently, probiotic drinks from fermented milk or probiotic yogurt dominate the global market (Market Analysis Report, 2020). One of the probiotic drink products that has been widely developed and is a good product as a carrier for probiotics is yogurt. Yogurt is a dairy product that is widely consumed throughout the world. In recent years, yogurt production has increased significantly, namely in 2015 it reached 35.5 million tons with a growth of more than 20% over the last 5 years. In Asia, yogurt production has also increased significantly, reaching 9.4 million tons with growth of more than 36% over the last 5 years (World Yogurt Market Report, 2016).
Probiotic drinks are typically made with cow's milk. However, milk from different sources has been frequently used to manufacture probiotic drinks, including goat's milk, sheep's milk, buffalo's milk, horse's milk, and camel's milk (Balthazar et al., 2016; Marnianti et al., 2021; De Santis et al., 2019; Adane Shegaw et al., 2020;
Sinamo et al., 2020). Horse milk is a type of milk that can be utilized in producing probiotic drinks since it has excellent composition and health benefits. The interest in horse milk processed products stems from its limited utilization, particularly in Indonesia. Furthermore,
fermented products from horse's milk are likely to boost the supply of dairy products with higher nutritious content. Horse milk is also an excellent substrate for the growth of probiotic bacteria.
Sumbawa wild horse milk is pure milk produced from native Sumbawa horses that are in the lactation period and breed naturally in grasslands (Yulianto and Saputri, 2017). Sumbawa wild horse milk is part of the people of West Nusa Tenggara's cultural heritage and ancestral traditions, especially on Sumbawa Island. Horse milk is considered a minor dairy product that has been gradually gaining popularity due to the socio-economic role of horses in the geographic areas where they live. It is known for its high nutritional value and health benefits (Prastyowati, 2021). In Central Asia, horse milk is a type of milk that has long been consumed and is believed to have health benefits.
One of the fermented horse’s milk products usually consumed is koumiss, or in Mongolia, it is called Airag. Koumiss or Airag is horse milk fermented using lactic acid bacteria (LAB) and yeast, which has sour and slightly alcoholic characteristics (Choi, 2016; Kondybayev et al., 2021;
Simonenko and Begunova, 2021). Based on research results from Suharto, (2019), it is known that Sumbawa wild horse milk contains 5.52% carbohydrates, 1.15%
protein, 1.14% fat, 91.53% water, 0.39%
ash, 0.04% crude fiber, and 3.11% total sugar. Sumbawa wild horse milk has a good shelf life, remaining stable at room temperature for up to 5 months without processing. This preservation ability is due to natural antimicrobial compounds and antimicrobials associated with lactic acid bacteria (LAB) found in the milk (Hermawati, 2005).
*Corresponding author:
Ahmad Sulaeman
Email: [email protected]
Department of Community Nutrition, Faculty of Human Ecology, IPB University, Bogor 16680, Indonesia
How to cite:
Fajriani, L. N., Sulaeman, A., & Setiawan, B.
(2023). Physicochemical and Sensory Quality of Probiotic Drink from Sumbawa Wild Horse Milk.
Jurnal Ilmu dan Teknologi Hasil Ternak (JITEK), 18 (3), 189-202
Despite its high nutritional content and good quality, Sumbawa wild horse milk remains relatively unpopular compared to cow's milk and other milk varieties, resulting in its limited utilization. The low acceptance of the product may be attributed to its inferior sensory quality resulting from natural fermentation (Sofiyatin and Widiada, 2018). The process of natural fermentation can lead to inconsistency in the quality and flavor of the product, as well as the risk of microbial contamination, which may include harmful pathogens or result in food spoiling (Suharto, 2019). The characteristics of Sumbawa wild horse milk are its thin consistency, sweet taste, and white color. However, during the whole process of fermentation, Sumbawa wild horse milk has a yellowish-white color, has a liquid consistency, and produces a sour aroma and taste.
Several studies related to horse milk and processed horse milk products such as yogurt have low sensory quality due to natural fermentation, so their acceptability is low due to they have a strong, distinctive taste and smell, so they have yet to be widely developed (Murti et al., 2015; Sofiyatin and Widiada, 2018; Marnianti et al., 2021). One way to increase the utilization and diversification of products from Sumbawa wild horse milk to become a better alternative product choice for the community than other commercial products is to develop it into a probiotic drink.
One factor that influences the quality of probiotic drinks is the type of starter.
Lactobacillus rhamnosus, classified as a probiotic bacteria, can enhance the nutritional content and sensory characteristics of probiotic drinks, making it a viable starter culture. According to the findings of Nuraida et al., (2014), yogurt made with L. rhamnosus R23 starter had the highest total LAB, 9.38 x 109 CFU/ml, a pH range of 3.68 to 3.82, and the panelists preferred its taste, smell, texture, and overall quality. The use of Lactobacillus bulgaricus and Streptococcus thermophilus is based on the fact that they are commonly used in
making commercial yogurt. However, these two bacteria are not probiotic bacteria, so they cannot survive in the digestive tract.
The aroma of yogurt fermented with L.
rhamnosus is similar in quality to that of yogurt fermented with standard culture (Innocente et al., 2016). According to De Santis et al., (2019), adding probiotic bacteria can improve the acceptability and sensory characteristics of fermented goat milk.
Based on the findings above, adding a probiotic starter is expected to improve product quality regarding physicochemical, total product LAB, sensory characteristics, and product acceptability. This study aims to develop a probiotic drink product from Sumbawa wild horse milk using different types of starters and then analyze the product's physicochemical, total LAB, sensory characteristics, and acceptability based on Indonesian National Standard 2981:2009.
MATERIALS AND METHODS
Time and location of research
This research was conducted at the Microbiology Laboratory, Food Biochemistry Laboratory, Bioprocess Engineering Laboratory, Faculty of Food Technology and Agroindustry, University of Mataram, West Nusa Tenggara.
Organoleptic test and Quantitative Descriptive Analysis (QDA) were carried out in the Organoleptic Laboratory, Department of Community Nutrition, Faculty of Human Ecology, IPB University, Bogor.
Research materials and methods
The research used an experimental design carried out in the laboratory, namely a Completely Randomized Design (CRD) with two treatments namely, different types of starters P1= Lactobacillus rhamnosus and P2= Lactobacillus bulgaricus and Streptococcus thermophilus. The materials used in this study were Sumbawa wild horse milk which is obtained directly from breeders residing in the Penyaring village,
North Moyo District of Sumbawa Regency, West Nusa Tenggara, Indonesia. Other ingredients consist of skim milk powder (Indoprima, Indonesia), honey (Mbojo Honey, Bima, Indonesia), and bacteria Lactobacillus rhamnosus FNCC 0052, Lactobacillus bulgaricus FNCC 0472, Streptococcus thermophilus FNCC 0040 were obtained from Center for Food and Nutrition Studies, Gadjah Mada University (UGM), Indonesia.
The equipment used in the manufacture and processing of Sumbawa wild horse milk probiotic drink is:
autoclave, incubator, water bath, homogenizer, aluminium foil, erlenmeyer, bottles, laminar flow.
Starter culture preparation
Making de Man Rogosa Sharpe Broth (MRSB) media
This process is adapted from (Marnianti et al., 2021). 2.34 g MRSB is dissolved in 45 mL of water (Oxoid). Then, the autoclave was used to sterilize the items for 5 minutes at 121°C.
Starter preparation
This process is adapted from (Marnianti et al., 2021). Lactobacillus rhamnosus FNCC 0052, Lactobacillus bulgaricus FNCC 0472, and Streptococcus thermophilus FNCC 0040 bacteria were obtained from Center for Food and Nutrition Studies, Gadjah Mada University (UGM), Indonesia. One loop of each stock culture was taken and inserted into the MRSB medium using a loop needle. After that, it was incubated in an incubator at 37°C for 48 hours.
Mother culture preparation
This process is adapted from (Marnianti et al., 2021). 200 mL of UHT milk (Greenfields) was mixed with 5% skim milk (Indoprima, Indonesia). Next, the mixture was pasteurized for 20 minutes at 65°C. The temperature was then lowered to 37°C. Inoculate pasteurized milk with 5% of each starter culture. After that, it was incubated for 48 hours in an incubator that is set to 37°C.
Ready-to-use culture preparation
This process is adapted from (Marnianti et al., 2021). 100 ml Sumbawa wild horse milk was mixed with 5% skim milk (Indoprima, Indonesia). Next, the mixture was pasteurized in a water bath at 65°C for 20 minutes. The temperature was then lowered to 37°C. In pasteurized Sumbawa wild horse milk, inoculate 5% of the mother cultures of L. rhamnosus FNCC 0052, L. bulgaricus FNCC 0472, and S.
thermophilus FNCC 0040. The next step was to incubate the samples in an incubator at 37°C for 48 hours.
The process of making probiotic drink from Sumbawa wild horse milk
The present study involves modifying the process based on Raut et al., (2015) to develop a probiotic drink using Sumbawa wild horse milk. The primary ingredient utilized in this context is Sumbawa wild horse milk, which is obtained directly from breeders residing in the Penyaring village, North Moyo District of Sumbawa Regency, West Nusa Tenggara, Indonesia. The milk obtained from Sumbawa wild horses exhibits distinct characteristics, including a sweet taste, a white coloration, a watery consistency, and a pH level of 6.49. A total volume of 500 ml of Sumbawa wild horse milk was prepared.
Then, it was mixed with 5% of skim milk (Indoprima, Indonesia). In addition, homogenization was conducted using a homogenizer at room temperature, operating at a rotational speed of 4000 revolutions per minute (rpm) for 10 minutes. The next step is pasteurization, in which a water bath was used at 65°C for 20 minutes. The temperature was lowered to 37°C. A 4%
ready-to-use culture was prepared for inoculation based on the treatments, P1 (L.
rhamnosus) with ratios 100% and P2 (L.
bulgaricus and S. thermophilus) with ratios 50%:50%. Afterward, the mixture was incubated at 37°C for 42 hours. The incubation time of 42 hours is based on preliminary research, in which the incubation time of 42 hours reached the pH of the probiotic drinks, and curd was
formed. The probiotic drink that underwent incubation was supplemented with 1%
honey (Mbojo Honey, Bima, Indonesia) and then stored below 10°C.
Variable Test
pH according to according to (AOAC, 2005)
The pH of the Sumbawa wild horse milk probiotic drink was measured using a pH meter (SCHOTT handy lab pH 11). The process of measurements starts by first calibrating the pH meter using a buffer.
Measurements were conducted by immersing the pH meter electrode into the sample and allowing sufficient time for the number on the pH meter to stabilize. The electrode was rewashed using distilled water prior to the next measurement. pH measurements were carried out in two repetitions.
Total titratable acidity analysis (%) according to (Nielsen, 2017)
Three drops of phenolphthalein indicator were applied to 10 ml of the sample. After that, the sample was titrated using a 0.1 N sodium hydroxide (NaOH) solution until a noticeable color transition to pink occurred. It was ensured that the color remained consistent throughout the homogenization process without any eventual disappearance. Total titratable
acidity measurements were carried out in two repetitions.
Total dissolved solid (°Bx) according to (AOAC, 2005)
The sample is dropped into the prism of the refractometer, and the measurement of °Bx is obtained. Total dissolved solid measurements were carried out in two repetitions.
Viscosity (mPa.s) according to (AOAC, 2005)
The viscosity was measured with a digital viscometer (Uchen). The spindle used is spindle number 1 at a speed of 60 rpm. The viscosity value can be immediately observed on a digital viscometer, expressed in millipascal-second units (mPa.s).
Viscosity measurements were carried out in two repetitions.
Determination of syneresis (%) according to (AOAC, 2005)
The centrifuge tube was weighed.
Then, a 15 g sample was added. Following the procedure above, centrifugation was performed with a centrifuge operating at 1500 rpm for 20 minutes. Furthermore, the liquid containing the precipitate was separated through filter paper. The resulting solid was later weighed, and the syneresis process is calculated.
Syneresis (%) = (𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡−𝑠𝑒𝑑𝑖𝑚𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡)
𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡 x 100%
Total lactic acid bacteria (LAB) (CFU/ml) according to (Pelczar and Chan, 2007)
The dilution buffer was prepared.
Dilution 1 (P-1) was made by adding 1 ml of the sample to 9 ml of buffer. To make dilution 2 (P-2), 1 ml of dilution 1 was added to 9 ml of buffer. Dilution was carried out up to P-9 (dilution 9). The final three dilutions, dilutions 7 (P-7), 8 (P-8), and 9 (P-9) were then taken and inoculated on de Man Rogosa Sharpe Agar (MRSA) media. It was then incubated in an incubator for 48 hours at 37°C. Next, the number of colonies is counted.
Organoleptic tests according to Indonesian National Standard or SNI-01- 2346-2006 (BSN, 2006)
The organoleptic test involved 35 semi-trained panelists who had either participated in or received material related to organoleptic tests. Hedonic liking tests with 9 rating scales (1= dislike extremely, 2=
dislike very much, 3= dislike, 4= dislike slightly, 5= neither like nor dislike, 6= like slightly, 7= like, 8= like very much, 9= like extremely) are included in organoleptic tests. Color, smell, texture, taste, mouthfeel, aftertaste, and overall were the sensory attributes tested.
Quantitative Descriptive Analysis (QDA) according to (Adawiyah et al., 2019;
García‐Gómez et al., 2019; Hunaefi et al., 2019)
The QDA test involved 8 trained panelists who had completed sensory attribute training and testing. The eight trained panelists are IPB Postgraduate Nutrition Science students who have completed the selection and training process. Previously trained panelists were chosen based on their skill and sensory acuity. Panelists were trained on basic tastes, smells, and textures, as well as how to identify the characteristics of the test samples. Following that, the product's sensory characteristics were discovered through a Focus Group Discussion (FGD) held before the QDA test to obtain panelist agreement on the sample characteristics. At the QDA test stage, each product's sensory attributes will be evaluated based on the previous FGD. All attributes will be reviewed by the panelists, ranking them from 1 to 10.
Statistical analysis
The research data were analyzed using Microsoft Excel 2019 and IBM Statistical Program Social Sciences (SPSS) version 23.0 (SPSS Inc., USA), specifically the independent T-test different test with a significant difference or p-value significance less than (p<0, 05).
RESULTS AND DISCUSSION
pH
Table 1 shows the physicochemical characteristics and total LAB of the Sumbawa wild horse milk probiotic drink.
The physicochemical characteristics were analyzed, including pH, total titratable acidity, total dissolved solid, viscosity, syneresis, and total LAB. The pH value is an indicator of the acidity level. Based on statistical analysis, it shows significant results which means that different types of starters influence reducing pH. The pH of the Sumbawa wild horse milk probiotic drink ranges from 4.09±0.01 to 4.35±0.01.
These findings suggest that the pH of fresh milk (6.49) decreases due to the fermentation process. One of the characteristics of fermentation by LAB is the production of various organic acids.
Organic acids in fermented dairy products became indicators of the metabolic activity of bacteria (Melia et al., 2021). The LAB fermentation process breaks lactose into lactic acid and produces other components because bacteria have different metabolic processes. L. rhamnosus is a heterofermentative bacterium, which, during the fermentation process, not only produces lactic acid but also produces ethanol, carbon dioxide, and acetic acid (Chen et al., 2017; Valik et al., 2008).
Meanwhile, L. bulgaricus and S.
thermophilus are homofermentative bacteria, that, during the fermentation process only produce lactic acid, so that the pH decreased more quickly as indicated by the P2 probiotic drink having a lower pH (Bostan et al., 2017; Sfakianakis and Tzia, 2014). Szajnar et al., (2021) reported that yogurt with L. rhamnosus had a higher pH than L. acidophilus which is homofermentative bacterium. Additionally, using more than one type of bacteria that can lower pH through a faster fermentation process can also result in a significant drop in pH. The pH of the probiotic drink P2 (4.09±0.01), which is lower than P1 (4.35±0.01), demonstrates this result. The results of this study are in line with the findings of Hassan and Amjad (2015), who found that the average pH of yogurt with L.
acidophilus was 4.29, while Yang and Yoon (2022) reported that Greek yogurt with L.
bulgaricus and S. thermophilus had a pH of 4.16.
Total titratable acidity
Table 1 shows that total titratable acidity is not significantly different. It means different types of starters do not affect the total titratable acidity. Total titratable acidity is the accumulation of total acid during fermentation. In Sumbawa wild horse milk probiotic drinks, the total titratable acidity value ranges from
0.55±0.07% to 0.61±0.02%. The total titratable acidity in P1 probiotic drink is lower (0.55±0.07%) than in P2 probiotic drink (0,61±0.02%). This is due to the lower pH of the P2 probiotic drink. The higher total titratable acidity in P2 probiotic drink indicates a higher acidity level, which is consistent with the lower pH. This finding is consistent with the findings of Rossi et al., (2021) who discovered that total titratable acidity levels are higher when pH is lower.
Yogurt with L. plantarum and S.
thermophilus starts has total titratable acidity levels between 0.63% and 0.99%.
One factor that influences the total titratable acidity is the amount of sugar. Sandra et al., (2019) reported that the greater the amount of sugar, the greater the activity of LAB, so more sugar is converted into lactic acid, which causes the pH to decrease and the total acid to increase. Besides that, Melia et al., (2021) stated that fermentation time affects the increase in lactic acid. The longer the fermentation, the production of lactic acid increase because more lactose is hydrolyzed into lactic acid. The results of this study fulfilled the total titratable acidity quality requirements for yogurt based on SNI 2981:2009 (Badan Standarisasi Nasional, 2009), namely a minimum of 0.5- 2.0%.
Total dissolved solids
In general, the total dissolved solids is a measurement utilized to determine the quantity of materials that have dissolved in a solution. When it comes to lactose, total dissolved solids can be used to figure out how much sugar is in a substance (Rizqiati et al., 2021). The Sumbawa wild horse milk probiotic drinks contain between 11.95±0.14°Bx and 13.10±0.07°Bx of total dissolved solids. Total dissolved solids differed significantly and generally decreased. Consistent with the findings of Rizqiati et al., (2021), the total dissolved solids content of buffalo milk kefir ranges from 4.00 to 4.94°Bx and decreases as the concentration of kefir starter increases. It's possible that the change happened because lactose broke down into galactose and
glucose, which then turned into lactic acid.
A drop in sugar levels can be a sign that microorganisms are active.
Microorganisms can grow best when there is enough sugar in the solution for it to break down, which lowers the percentage of sugar in the solution. The change may occur due to the degradation of lactose into galactose and glucose, which results in the formation of lactic acid as the final product.
A decrease in sugar levels can indicate the presence of microorganism activity.
Microorganisms can grow best when there is enough sugar in the solution for it to break down, which lowers the concentration of sugar in the solution. In addition to these substances, total dissolved solids may also include proteins, pigments, lactic acid, and organic acids (Ismawati, 2016). Based on the findings of Rossi et al., (2021) the total dissolved solid value of full-cream milk yogurt ranges from 9.63 to 10.40%. The results of this study fulfilled the solid non fat quality requirements for yogurt based on SNI 2981:2009 (Badan Standarisasi Nasional, 2009), namely a minimum of 8.2%.
Viscosity
The viscosity of the fresh Sumbawa wild horse milk increases after going through a fermentation process. The viscosity of the Sumbawa wild horse milk probiotic drink ranges from 57.00±2.82 mPa.s to 78.00±2.82 mPa.s. Different types of starters have a significant effect on viscosity. The rise in viscosity can be attributed to the production of exopolysaccharides (EPS) by bacteria during the fermentation process. The viscosity of the probiotic drink P1 is greater than that of the probiotic drink P2 because probiotic drink P1 contains L. rhamnosus that can produce more exopolysaccharides (EPS) than other LABs. Meanwhile, L.
bulgaricus and S. thermophilus can produce EPS in glucose-rich media. EPS is a polysaccharide produced by microbial cells that has a variety of roles in improving yogurt characteristics, including texture (Muncan et al., 2020).
Aside from the fermentation process, viscosity can be influenced by adding skim milk, which increases the total dissolved solids. This can also occur due to the probiotic drink P1 having a higher concentration of dissolved solids, resulting in increased viscosity. Suharto et al., (2016) found that the incorporation of rosella into goat's milk yogurt resulted in increased viscosity, ranging from 127 mPa.s to 243 mPa.s. This is because goat's milk has higher total solids and fat, so the viscosity of goat's milk yogurt is higher than cow's milk.
Sumbawa wild horse milk probiotic drinks have a lower viscosity than Suharto’s research results. The protein content can influence it. Protein content is one of the factors that plays an important role in coagulum formation so that the product’s viscosity is directly proportional to the level of protein present. Sumbawa wild horse milk probiotic drinks have a low viscosity because horse milk has a higher whey protein content than casein. The low casein content in horse milk causes the coagulation process during fermentation to be weak to
have a more liquid consistency (Kondybayev et al., 2021).
Syneresis
Syneresis is a condition when two phases are separated, specifically water and whey (Paz-Díaz et al., 2021). The probiotic drink syneresis in Sumbawa wild horse milk ranged from 74.66±5.61% to 80.65±4.19%.
In general, the increase in syneresis is affected by total dissolved solids. Based on the research results, it is known that the higher the total dissolved solid the lower the syneresis and the higher the viscosity.
Syneresis is likewise negatively related to viscosity, with low syneresis indicating high viscosity (Wati et al., 2018). Adane Shegaw et al., (2020) reported that camel milk yogurt without adding EPS had the highest syneresis of 71% compared to yogurt with EPS addition, which was 67%. Furthermore, changes in acidity might produce higher syneresis. The P2 probiotic drink has a pH of 4.09, indicating strong acidity. This high acidity in yogurt can lead to an increase in syneresis (Yilmaz-Ersan and Kurdal, 2014).
Table 1. Physicochemical characteristics and total LAB of probiotic drink from Sumbawa wild horse milk
Properties Value
Significance
P1 P2
pH 4,35±0,01 4,09±0,01 0,003*
Total Titratable Acidity (%) 0,55±0,07 0,61±0,02 0,381
Total Dissolved Solids (°Bx) 13,10±0,14 11,95±0,07 0,009*
Viscosity (mPa.s) 78,00±2,82 57,00±2,82 0,018*
Syneresis (%) 74,66±5,61 80,65±4,19 0,350
Total LAB (CFU/ml) 3,42x109±0,49 2,56x109±0.01 0,135
P1= L. rhamnosus, P2= L. bulgaricus and S. thermophilus. *p value <0,05 (significance)
Total lactic acid bacteria (LAB)
Sumbawa wild horse milk naturally contains LAB. L. acidophilus, L. brevis, and L. rhamnosus are prospective probiotic candidates isolated from Sumbawa wild horse milk (Antara et al., 2009; Sujaya, 2008; Shi et al., 2012; Widiada, 2021). The total LAB of fresh Sumbawa wild horse milk was 2.9x108 CFU/ml. The total LAB increased after Sumbawa wild horse milk was converted into a probiotic drink. Based on statistical analysis, different types of
starters do not have significant effect on total LAB. The probiotic drinks treated with P1 had a significantly higher total LAB count of 3.42x109 CFU/ml compared to the probiotic drinks treated with P2, which had a count of 2.56x109 CFU/ml. Paramita et al. (2017) reported that yogurt containing L.
rhamnosus A6 has a total LAB count of 3.74x106 CFU/ml. In line with the research of Nuraida et al., (2014) yogurt containing L. rhamnosus R23 had the highest total LAB, specifically 9.38 log CFU/ml. This
could be because L. rhamnosus grows well as a sole starter in yogurt production. The minimum total LAB specified in SNI 2981:2009 (Badan Standarisasi Nasional, 2009) for yogurt products is 107 CFU/ml.
Based on the Food Drug Administration (FDA), the minimum strain of microorganisms for yogurt is 106-108 CFU/ml. Meanwhile, based on BPOM Regulation Number 1 of 2022, the minimum number of live probiotic microorganism strains is 108 CFU/ml per serving. The total LAB range of the probiotic drink was 2.56x109 to 3.42x109 CFU/ml, indicating that the probiotic drink made from Sumbawa wild horse milk met the SNI, FDA, and BPOM requirements for the total LAB of yogurt.
Organoleptic hedonic test probiotic drink from Sumbawa wild horse milk
Table 2 depicts the hedonic characteristics of the Sumbawa wild horse milk probiotic drink. The T difference test's statistical results revealed that it was insignificant for any sensory attributes. This suggests that different types of starter treatment do not affect the overall sensory attributes of the Sumbawa wild horse milk
probiotic drink. This is because the components used are all the same; the only difference is the type of starter used.
The Sumbawa wild horse milk probiotic drink has an average overall preference score of 5.29±1.44 to 5.39±1.04. The like results suggest that the Sumbawa wild horse milk probiotic drink with commercial bacteria, specifically L. bulgaricus and S.
thermophilus (P2), has a preference that is higher in various sensory attributes, including color (7.08±1.46), taste (5.31±1.19), mouthfeel (5.13±1.29), and aftertaste (5.03±1.17). The smell (5.46±1.67) and texture (6.00±1.57) attributes of the Sumbawa wild horse milk probiotic drink with the probiotic bacteria L.
rhamnosus (P1) tend to be higher. The like test of taste preference results aligns with research by Yang and Yoon, (2022) that Greek yogurt has the highest preference for taste (5.03). Most panelists agreed that the Sumbawa wild horse milk probiotic drink has a sour taste and a strong, distinctive smell. This could be because the raw material used for Sumbawa wild horse milk has a strong, distinct smell and taste, which may have reduced the panelists' preferences.
Table 2. Hedonic preference level for the probiotic drink from Sumbawa wild horse milk
Attribute Hedonic Score
Significance
P1 P2
Color 7,01±1.29 7,08±1,46 0,843
Smell 5,46±1,67 5,37±1,69 0,837
Texture 6,00±1,57 5,78±1,56 0,559
Taste 5,07±1,14 5,31±1,19 0,383
Mouthfeel 5,00±1,35 5,13±1,29 0,687
Aftertaste 4,84±1,32 5.03±1,17 0,530
Overall 5,29±1,44 5,39±1,04 0,727
P1= L. rhamnosus. P2= L. bulgaricus, and S. thermophilus. 1= dislike extremely, 2= dislike very much, 3=
dislike, 4= dislike slightly, 5= neither like nor dislike, 6= like slightly, 7= like, 8= like very much, 9= like extremely.
Quantitative Descriptive Analysis (QDA) probiotic drink from Sumbawa wild horse milk
Quantitative Descriptive Analysis is a tool used to measure and optimize the sensory quality attributes of a product. The QDA method is carried out based on the principle of the panelists’ ability to measure specific quality attributes of a product
(Hunaefi and Ulfah, 2019; Puri et al., 2016;
Rahmawati et al., 2015). The sensory qualities obtained by QDA are as follows:
appearance (color and smoothness), smell (skunky and sour), texture (viscosity), taste (skunky, sour, bitter, sweet), and mouthfeel (skunky). The QDA results show that the probiotic drink P2 (L. bulgaricus and S.
thermophilus) has a whiter color and a more
sour taste than P1 (L. rhamnosus). The whiter color is produced by casein without carotene, while the slightly yellow color is caused by fat globules in milk (Khoiriyah and Fatchiyah, 2013).
The panelists preferred a more sour taste since it can mask the skunky taste. In contrast to Yang and Yoon, (2022) findings, Greek yogurt containing L. bulgaricus and S. thermophilus starters had the lowest preference score for acidity. The probiotic
drink P1 (L. rhamnosus) appears smoother and thicker. The sour smell and skunky taste attribute of Sumbawa wild horse milk is more prominent in probiotic drink P1, decreasing the panelists' liking. Apart from that, probiotic drink P1 has a slightly bitter and sweet taste, influencing the liking for the aftertaste.
The addition of 1% honey is insufficient to mask the characteristic smell and taste of Sumbawa wild horse milk.
Figure 1. Intensity of sensory quality of Sumbawa wild horse milk probiotic drink made from different types of starter. This study used L. rhamnosus (P1), L. bulgaricus and S.
thermophilus (P2) CONCLUSION
Different types of starters substantially enhance the physicochemical characteristics, total LAB, and sensory quality of probiotic drinks made from Sumbawa wild horse milk. According to SNI 2981:2009, all probiotic drinks formulations met the total LAB requirements for yogurt. Based on the physicochemical characteristics probiotic drink P1 can improve the physicochemical product, and organoleptic hedonic test, the probiotic drink with commercial starters (P2) had the highest sensory acceptability with a whiter color and more sour taste.
Disclosure and conflict of interest
The authors declare that they have no competing interests.
ACKNOWLEDGMENT
We present special thanks for all the people who have helped this research.
REFERENCES
Shegaw, A., Ipsen, R., Kurtu, M. Y., Eshetu, M., Hansen, E. B., Hailu, Y., Waktola, A., & Dashe, D. (2020). Development of Yogurt from Camel Milk Using Exopolysaccharide Producing Lactic Acid Bacteria. Eth. J. Anim. Prod., 20(1), 29–43.
Adawiyah, D. R., Azis, M. A., Ramadhani, A. S., & Chueamchaitrakun, P.
(2019). Perbandingan profil sensori teh hijau menggunakan metode analisis deskripsi kuantitatif dan cata (check-all-that-apply). Jurnal
Teknologi dan Industri Pangan, 30(2), 161–172. https://doi.org/10.6066/jtip.
2019.30.2.161
Antara, N. S., Dibia, I. N., & Aryanta, W. R.
(2009). Karakterisasi Bakteri Asam Laktat yang diisolasi dari susu kuda Bima. Agritech: Jurnal Fakultas Teknologi Pertanian UGM, 29(1), 89873.
AOAC. (2005). Official Methods of Analysis of International 18th Ed.
Badan Standarisasi Nasional. (2009). SNI 2981:2009 tentang Yogurt.
Balthazar, C. F., Conte Júnior, C. A., Moraes, J., Costa, M. P., Raices, R. S.
L., Franco, R. M., Cruz, A. G., &
Silva, A. C. O. (2016).
Physicochemical evaluation of sheep milk yogurts containing different levels of inulin. Journal of Dairy Science, 99(6), 4160–4168. https://
doi.org/10.3168/jds.2015-10072 Bostan, K., Unver Alcay, A., Yalçin, S.,
Eren Vapur, U., & Nizamlioglu, M.
(2017). Identification and characterization of lactic acid bacteria isolated from traditional cone yoghurt.
Food Science and Biotechnology, 26(6), 1625–1632. https://doi.org/10.
1007/s10068-017-0222-z
BPOM. (2022). Peraturan badan pengawas obat dan makanan nomor 1 tahun 2022 tentang pengawasan klaim pada label dan iklan pangan olahan.
BSN. (2006). Petunjuk pengujian sensoris dan sensori. Standar Nasional Indonesia 01-2346-2006.
Chen, C., Zhao, S., Hao, G., Yu, H., Tian, H., & Zhao, G. (2017). Role of lactic acid bacteria on the yogurt flavour: A review. International Journal of Food Properties, 20(sup1), S316–S330.
https://doi.org/10.1080/10942912.201 7.1295988
Choi, S.-H. (2016). Characterization of airag collected in Ulaanbaatar, Mongolia with emphasis on isolated lactic acid bacteria. Journal of Animal Science and Technology, 58(1), 10. https://
doi.org/10.1186/s40781-016-0090-8
De Santis, D., Giacinti, G., Chemello, G., &
Frangipane, M. T. (2019).
Improvement of the sensory characteristics of goat milk yogurt.
Journal of Food Science, 84(8), 2289–
2296. https://doi.org/10.1111/1750- 3841.14692
FAO/WHO. (2002). Health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria, a joint FAO/WHO expert consultation. Cordoba, Argentina, 1-4 October 2001.
FDA. (2008). Food labelling guide:
appendix B: additional requirements for nutrient content claims.
García‐Gómez, B., Romero‐Rodríguez, Á., Vázquez‐Odériz, L., Muñoz‐Ferreiro, N., & Vázquez, M. (2019). Sensory quality and consumer acceptance of skim yoghurt produced with transglutaminase at pilot plant scale.
International Journal of Dairy Technology, 72(3), 388–394. https://
doi.org/10.1111/1471-0307.12595 Hassan, A., & Amjad, I. (2015). Nutritional
evaluation of yoghurt prepared by different starter cultures and their physiochemical analysis during storage. Advanced Journal of Microbiology Research.
Hermawati, D. (2005). Kajian aktivitas dan karakterisasi senyawa antimikroba dari susu kuda Sumbawa. Disertasi.
Hunaefi, D., Khairunnisa, W., Sholehuddin, Z., & Adawiyah, D. (2019). Sensory profile of commercial coffee products using QDA (Quantitative Descriptive Analysis), flash profile, and CATA (Check-All-That-Apply) methods.
Proceedings of the 2nd SEAFAST International Seminar, 20–30. https://
doi.org/10.5220/0009977500200030 Hunaefi, D., & Ulfah, F. (2019). Pendugaan
umur simpan produk pastry dengan quantitative descriptive analysis (QDA) dan metode arrhenius. Jurnal Mutu Pangan: Indonesian Journal of Food Quality, 6(2), 72–78. https://
doi.org/10.29244/jmpi.2019.6.72
Innocente, N., Biasutti, M., Rita, F., Brichese, R., Comi, G., & Iacumin, L.
(2016). Effect of indigenous Lactobacillus rhamnosus isolated from bovine milk on microbiological characteristics and aromatic profile of traditional yogurt. LWT - Food Science and Technology, 66, 158–
164. https://doi.org/10.1016/j.lwt.20 15.10.031
Ismawati, N. (2016). Nilai ph, total padatan terlarut, dan sifat sensoris yoghurt dengan penambahan ekstrak bit (beta vulgaris l.). Jurnal Aplikasi Teknologi Pangan, 5(3). https://doi.org/10.17728 /jatp.181
Khoiriyah, L. K., & Fatchiyah, F. (2013).
Karakter biokimia dan profil protein yogurt kambing PE difermentasi bakteri asam laktat (BAL). The Journal of Experimental Life Sciences, 3(1), 1–6. https://doi.org/
10.21776/ub.jels.2013.003.01.01 Kondybayev, A., Loiseau, G., Achir, N.,
Mestres, C., & Konuspayeva, G.
(2021). Fermented mare milk product (Qymyz, Koumiss). International Dairy Journal, 119, 105065.
https://doi.org/10.1016/j.idairyj.2021.
105065
Market Analysis Report. (2020). Probiotic drink market size, share and trends analysis report by product (dairy- based, plant-based), by distribution channel (offline, online), by region, and segment forecast, 2020-2070.
Cicilia, S., Nazaruddin, N., & Marnianti, S.
S. (2021). The quality of yoghurt wild horse milk with the addition of sweet wood extract in various concentrations. Pro Food, 7(1), 773- 784.
Melia, S., Juliyarsi, I., Fitri Kurnia, Y., Endra Pratama, Y., & Azahra, H.
(2021). Examination of Titratable acidity, pH, total lactic acid bacteria and sensory properties in whey fermented with probiotic pediococcus acidilactic BK01. Advances in Animal and Veterinary Sciences, 10(1).
https://doi.org/10.17582/journal.aavs/
2022/10.1.114.119
Muncan, J., Tei, K., & Tsenkova, R. (2020).
Real-Time monitoring of yogurt fermentation process by aquaphotomics near-infrared spectroscopy. Sensors, 21(1), 177.
https://doi.org/10.3390/s21010177 Murti, T. W., Robiyati, E., Jundi, H. L.,
Ramadhani, F., Rustamadji, B., &
Suranindyah, Y. Y. (2015).
Development of fermented mare’s milk using mixed probiotic cultures.
Media Peternakan, 39(1), 9–13.
https://doi.org/10.5398/medpet.2016.
39.1.9
Nielsen, S. S. (2017). Food analysis laboratory manual. Springer International Publishing. https://doi.
org/10.1007/978-3-319-44127-6 Nuraida, L., Nurdin, Q., & Firlieyanti, A. S.
(2014). Pengembangan yoghurt berisi Lactobacillus rhamnosus dan Pediococcus pentosaceus dan Viabilitasnya selama penyimpanan.
Jurnal Mutu Pangan: Indonesian Journal of Food Quality, 1(1), 47-55.
Paramita, S., & Arihantana, P. S. (2017).
Studi potensi Lactobacillus rhamnosus A6 hasil isolasi dari air susu ibu sebagai starter dalam pembuatan yoghurt. Media Ilmiah Teknologi Pangan, 4(2), 103-112.
Paz-Díaz, H. J., Mora-Vergara, L. Z., Navarro-Arana, C. E., Navarro, A. S.,
& Pacheco-Valderrama, M. M.
(2021). Effect of process variables on the physicochemical and rheological properties of yogurt. Revista U.D.C.A Actualidad & Divulgación Científica, 24(1). https://doi.org/10.31910/rudca.
v24.n1.2021.1922
Pelczar, M, & Chan, R. C. (2007). Dasar- dasar Mikrobiologi.
Prastyowati, A. (2021). Sumbawa wild horse milk: production, usage, chemical compound, microbial community, and probiotics potency.
Indonesian Bulletin of Animal and Veterinary Sciences, 31(3), 147.
https://doi.org/10.14334/wartazoa.v3 1i3.2850
Puri, R., Khamrui, K., Khetra, Y., Malhotra, R., & Devraja, H. C. (2016).
Quantitative descriptive analysis and principal component analysis for sensory characterization of Indian milk product cham-cham. Journal of Food Science and Technology, 53(2), 1238–1246. https://doi.org/10.1007/
s13197-015-2089-4
Rahmawati, D., Andarwulan, N., & Lioe, H.
N. (2015). Identifikasi atribut rasa dan aroma mayonnaise dengan metode quantitative descriptive analysis (QDA). Jurnal Mutu Pangan, 2(2), 80- 87.
Raut, V., Sawant, P., Sawant, D., & Ingole, A. S. (2015). Studies on preparation of mango yoghurt drink. Asian Journal of Dairy and Food Research, 34(1), 13.
https://doi.org/10.5958/0976-0563.20 15.00003.2
Rizqiati, H., Nurwantoro, Susanti, S., Surya, K. R. A., & Prayoga, M. I. Y. (2021).
Chemical and microbiological properties of buffalo milk kefir with different starter concentrations. IOP Conference Series: Earth and Environmental Science, 803(1), 012045. https://doi.org/10.1088/1755- 1315/803/1/012045
Rossi, E., Restuhadi, F., Efendi, R., & Dewi, Y. K. (2021). Physicochemical and microbiological properties of yogurt made with microencapsulation probiotic starter during cold storage.
Biodiversitas Journal of Biological Diversity, 22(4). https://doi.org/10.13 057/biodiv/d220450
Sandra, A., Kurnia, Y. F., Sukma, A., &
Purwati, E. (2019). The chemical characteristics of yoghurt (Lactobacillus fermentum MGA40-6 and Streptococcus thermophilus) with additional puree from Senduduk fruit (Melastoma malabathricum, L.). In IOP Conference Series: Earth and Environmental Science (Vol. 287, No.
1, p. 012024). https://doi.org/10.1088/
1755-1315/287/1/012024
Sfakianakis, P., & Tzia, C. (2014).
Conventional and innovative processing of milk for yogurt manufacture; development of texture and flavor: a review. Foods, 3(1), 176–
193. https://doi.org/10.3390/foods301 0176
Shi, T., Nishiyama, K., Nakamata, K., Aryantini, N. P. D., Mikumo, D., Oda, Y., Yamamoto, Y., Mukai, T., Sujaya, I. N., Urashima, T., & Fukuda, K.
(2012). Isolation of potential probiotic Lactobacillus Rhamnosus strains from traditional fermented mare milk produced in Sumbawa Island of
Indonesia. Bioscience,
Biotechnology, and Biochemistry, 76(10), 1897–1903. https://doi.org/10.
1271/bbb.120385
Simonenko, E. S., & Begunova, A. V.
(2021). Development of fermented milk product based on mare milk and lactic microorganisms association.
Problems of Nutrition, 90(5), 115–
125. https://doi.org/10.33029/0042- 8833-2021-90-5-115-125
Sinamo, K. N., Hasan, F., & Hasanah, U.
(2020). Effect of concentration starter and fermentation time on the quality of yoghurt drink from buffalo milk.
Journal of Physics: Conference Series, 1542(1), 012070. https://doi.org/10.
1088/1742-6596/1542/1/012070 Sofiyatin R, W. I. (2018). Pengaruh
penambahan kacang merah terhadap sifat organoleptik produk minuman susu kuda sumbawa. Jurnal Kesehatan Prima, 12(1), 07–13.
Suharto, A. R. (2019). Dinamika populasi bakteri pada fermentasi alami susu kuda sumbawa melalui pendekatan metagenomik dan aspek fisikokimia.
Skripsi.
Suharto, E. L. S., Arief, I. I., & Taufik, E.
(2016). Quality and antioxidant activity of yogurt supplemented with roselle during cold storage. Media Peternakan, 39(2), 82–89. https://doi.
org/10.5398/medpet.2016.39.2.82
Sujaya, N., Ramona, Y., Widarini, N. P., Suariani, N. P., Dwipayanti, N. M. U., Nocianitri, K. A., & Nursini, N. W.
(2008). Isolasi dan karakterisasi bakteri asam laktat dari susu kuda Sumbawa. Jurnal Veteriner, 9(2), 52- 59.
Szajnar, K., Pawlos, M., & Znamirowska, A.
(2021). The effect of the addition of chokeberry fiber on the quality of sheep’s milk fermented by Lactobacillus rhamnosus and Lactobacillus acidophilus.
International Journal of Food Science.
https://doi.org/10.1155/2021/7928745 Liptáková, D., Valík, L., & MedveĎová, A.
(2008). Characterization of the growth of Lactobacillus rhamnosus GG in milk at suboptimal temperatures. J Food Nutr Res, 47, 60-67.
Vlasova, A. N., Kandasamy, S., Chattha, K.
S., Rajashekara, G., & Saif, L. J.
(2016). Comparison of probiotic lactobacilli and bifidobacteria effects, immune responses and rotavirus vaccines and infection in different host species. Veterinary Immunology and Immunopathology, 172, 72–84.
https://doi.org/10.1016/j.vetimm.201 6.01.003
Wati, A. M., Lin, M. J., & Radiati, L. E.
(2018). Physicochemical characteristic of fermented goat milk
added with different starters lactic acid bacteria. Jurnal Ilmu Dan Teknologi Hasil Ternak, 13(1), 54–62.
https://doi.org/10.21776/ub.jitek.2018 .013.01.6
Widiada, I. G. N. W. (2021). Karakterisasi fenotifik isolat bakteri asam laktat (bal) dari susu kuda liar bima. Jurnal Riset Kesehatan Poltekkes Depkes Bandung, 13(2), 304–309.
https://doi.org/10.34011/juriskesbdg.
v13i2.1867
World Yogurt Market Report. (2016). The World Yoghurt Market Report 2000- 2025.
Yang, S.-Y., & Yoon, K.-S. (2022). Effect of probiotic lactic acid bacteria (LAB) on the quality and safety of greek yogurt. Foods, 11(23), 3799. https://
doi.org/10.3390/foods11233799 Yilmaz-Ersan, L., & Kurdal, E. (2014). The
production of set-type-bio-yoghurt with commercial probiotic culture.
International Journal of Chemical Engineering and Applications, 5(5), 402–408. https://doi.org/10.7763/IJC EA.2014.V5.418
Yulianto, K., & Saputri, D. S. (2017).
Strategi peningkatan mutu susu kuda di kabupaten Sumbawa. Jurnal TAMBORA, 2(3). https://doi.org/10.
36761/jt.v2i3.169
