DOI: 10.21776/ub.jiip.2023.033.02.06 188
The Effect of Tamarind Seeds in Fermented Liquid Feed as Pig Feed on Nutrient and Tannin Content, Lactic Acid Bacteria Population, and Fiber
Fraction
Redempta Wea*1), Andy Yumina Ninu1), Maria Milka Nurak2), Bernadete Barek Koten2), and Yustus Serani Nong Mbeong3)
1) Animal Production Study Program, Department of Animal Husbandry, Kupang State Agricultural Polytechnic, Kupang 85011, Indonesia
2) Animal Feed Technology, Department of Animal Husbandry, Kupang State Agricultural Polytechnic, Kupang 85011, Indonesia
3) Kupang State Agricultural Polytechnic, Kupang 85011, Indonesia Submitted: 14 June 2023, Accepted: 04 August 2023
ABSTRACT: Feed is the biggest cost in the pig livestock business, therefore it is necessary to do processing using fermentation technology by utilizing tamarind seeds in the liquid feed.
This study aimed to determine the nutrient, tannins, and populations of lactic acid bacteria (LAB), and fiber fraction. This study was undertaken in May-July 2021. The study consisted of 4 treatments, namely; R0: Fermented liquid feed (FLF) of 0% tamarind seeds, R10: FLF of 10% tamarind seeds, R20: FLF of 20% tamarind seeds, and R30: FLF of 30% tamarind seeds.
This study was conducted on the experimental design using a Completely Randomized Designed (CRD). The study variables were nutrient content, tannin, LAB population, and fiber fraction. The results were analyzed quantitatively using Analysis of Variance (ANOVA), and the differences between treatments were further tested by DMRT. The results showed that the percentage of using tamarind seeds in a ratio up to 30% had a very significant effect (P<0.01) on the nutrient content (dry matter, crude protein, crude fiber, nitrogen-free extract (NFE), crude fat, and ash), anti-nutrient (tannin), fiber fraction (NDF, ADF, cellulose, hemicellulose, and lignin), but did not affect (P>0.05) the population of lactic acid bacteria. In conclusion, the use of tamarind seeds up to 30% in liquid feed increases nutrient content (dry matter, crude protein, crude fiber, nitrogen-free extract (NFE), and ash), anti-nutrient (tannin), and fiber fraction content (NDF, ADF, cellulose, hemicellulose, and lignin), but was able to reduce the crude fat content and had no impact on the population of lactic acid bacteria.
Keywords: Bioconversion; Pigs; Ration; Soluble carbohydrates; Tannins
*Corresponding Author: [email protected]
DOI: 10.21776/ub.jiip.2023.033.02.06 189 INTRODUCTIONS
Maintenance of pigs with good productivity can be produced if given quality feed. However, pig feed ingredients are generally conventional feed ingredients that compete with human needs. One of the unconventional feed ingredients that can be used is tamarind seeds. Wea et al (2018), the people of East Nusa Tenggara are accustomed to using tamarind seeds as pig feed, but their use in feeds has not been formulated properly so the performance of livestock production and reproduction is not optimal.
The results of the author's research indicate that the nutrient content of tamarind seeds is 92.19% dry matter, 14.93% crude protein, 6.75% crude fiber, 5.58% crude fat, 2.39% ash, 0.41% Ca, 0.07% P, and 5000.49 Kcal/kg of energy, but also has limitations in its use, namely hard seed coats and anti- nutrient (tannins), namely 2.47%. Tannins are classified as polyphenolic compounds with characteristics that can form complex compounds with other macromolecules so that they are not easily digested (Westendarp 2006). The limitations of using tamarind seeds can be overcome by using fermentation technology.
The processing of whole tamarind seeds is usually done as a single feed ingredient, while liquid feed fermentation is not widely used (Wea, Ninu, and Koten 2020). Missotten et al (2015) stated that fermented pig liquid feed is usually in a ratio of 1:1.5 – 1:4. According to research by Wea, Ninu, and Koten (2020), fermentation of liquid feed with a water ratio of 1:3 affects dry matter content but has no impact on crude protein, crude fat, crude fiber, nitrogen-free extract (NFE), ash, and anti- nutrient (tannins). Liquid feed made from whole tamarind seeds with a water ratio of 1:3 and a fermentation time of 14 days provides nutritional value and fiber fraction (increased hemicellulose and decreased
lignin) (Wea, Ninu, and Koten 2020). The results of a study by Wea and Koten (2013) showed that the use of fermented tamarind seeds up to 30% had no effect on the body composition of local pigs. Therefore, research was conducted on nutritional quality, tannins, lactic acid bacteria (LAB) content, and fiber fraction in liquid feed using different percentages of tamarind seeds. The aim is to assess nutritional quality, tannins, LAB, and fiber fraction using a 1:3 ratio of feed and water and 14 days of fermentation.
MATERIALS AND METHODS Research location
The research took place from May to July 2021 at the nutrition and animal feed laboratory, Animal Husbandry Department of the Agricultural Polytechnic of Kupang State.
Research materials
Feed ingredients such as rice bran, corn, soybean meal, and meat and bone meal are mixed with tamarind seeds. The tamarind seeds used are those that have been sorted from foreign objects such as stones and tamarind peel and have gone through a floating test process to determine the quality of the tamarind seeds. The number of tamarind seeds used was following the research treatment.
Then the tamarind seeds are mixed with other feed ingredients whose nutritional composition has been calculated (Tables 1 and 2). A mixture of feed ingredients with tamarind seeds mixed with water in a ratio of 1:3. Liquid feed is fermented for 14 days. After 14 days of liquid feeding, the population of lactic acid bacteria was analyzed in a fresh state. The fermented products were dried in an oven at 60°C for 48 hours, and the samples were prepared for analysis of nutrient and anti- nutrient (tannins) content.
DOI: 10.21776/ub.jiip.2023.033.02.06 190 Table 1. Nutrition Composition of Research Feed Ingredients
Feed ingredients Nutrition composition (%)
Dry Matter Crude Protein Crude Fat
Crude Fiber
Ca P ME
(Kcal/kg)
Corn 89 8.3 3.9 2.8 0.03 0.28 3420
Rice Bran 90 13.3 13 13.9 0.07 1.16 2850
MBM 93 51.5 10.9 5.6 9.99 4.98 2225
Soybean Meal 90 47.5 2.9 5.4 0.34 0.69 3380
Tamarind seeds * 84.87 14.19 5.58 6.75 0.41 0.08 3302
Note: NRC (1998), * Wea et al (2019), MBM = meat and bone meal, Ca = calcium, P = phosporus
Table 2. Composition and Nutrition Content of Research Feed
Feed ingredients Feed Composition (%)
R0 R1 R2 R3
Corn 53 49.5 47.5 40.5
Rice Bran 27 21.5 14.5 12.5
Meat and Bone Meal 8 10.2 13.5 11
Soybean Meal 12 8.8 4.5 6
Tamarind seeds 0 10 20 30
Nutrient content of feed:
Dry Matter (%) 89.71 89.30 88.90 88.39
Metabolizable energy (Kcal/kg) 3165.70 3160.24 3150.63 3179.50
Crude Protein (%) 17.81 17.82 17.80 17.80
Crude Fat (%) 6.80 6.65 6.46 6.25
Crude Fiber (%) 6.33 6.10 5.69 5.84
Ca (%) 0.87 1.12 1.47 1.26
P (%) 0.94 0.96 1.02 0.87
Note: the calculation results, Ca = calcium, P = phosporus
Research parameters
The observed parameters were sorghum nutrients production variables were nutrient content (dry matter, organic matter, crude protein, crude fiber, nitrogen- free extract (NFE), crude fat, and ash) and fiber fraction according to the AOAC procedure (2012), anti-nutrient (tannin), and lactic acid bacteria (LAB) will be analyzed the content of total LAB CFU/ml by diluting the sample sequentially, ie 101 - 108 (Fardiaz 1989).
Experimental design
The design used was a completely randomized design with 6 replications. The treatments consisted of R0 = fermented liquid feed without tamarind seeds, R1 = fermented liquid feed containing 10%
tamarind seeds, R2 = fermented liquid feed containing 20% tamarind seeds, and R3 =
fermented liquid feed containing 30%
tamarind seeds.
Data analysis
The results of the study will be analyzed quantitatively using the Analysis of Variance (ANOVA). The result with differences between the treatments was further tested with Duncan's test (Gomez and Gomez 2010).
RESULTS AND DISCUSSION
Fermentation technology carried out on liquid feed made from tamarind seeds with different percentages in the feed affects the nutrient and tannins content, but not affects lactic acid bacteria. The average effect of different percentages of tamarind seeds in the feed on the nutritional content, tannins, and LAB population, and is presented in Table 3.
DOI: 10.21776/ub.jiip.2023.033.02.06 191 Table 3. Nutrient content, anti-nutrient (tannins), and lactic acid bacteria in a fermented liquid
feed with different percentages of tamarind seeds
Nutrient content, % Without tamarind seeds
10% tamarind seeds
20% tamarind seeds
30% tamarind seeds
Dry matter 22.23±0.64 b 22.05±0.20 b 23.61±2.81 b 28.15±0.81 a Crude Protein 21.46±0.23 b 21.07±0.55 c 21.41±0.28 b 22.84±0.12 a
Crude fiber 9.38±0,41c 9.64±0,37bc 10.08±0,28b 11.22±0,55 a
NFE 44.87±0.81 b 44.79±0.79b 47.57±0.46 a 47.71±0.99 a
Crude fat 16.87±0.43a 17.31±0.59a 13.35±0.63b 10.27±0.37c
Ash 7.40±0.22 b 7.17±0.10 b 7.57±0.15 ab 7.93±0.66 a
Tannins 1.17±0.16b 1.22±0.01b 1.55±0.05a 1.59±0.04a
LAB (CFU/ml) 5.04 5.00 5.01 5.04
Note: a, b, c, means in the same rows with different superscripts differ significantly (p<0.05); NFE = nitrogen-free extract; R0: Fermented liquid feed of 0% tamarind seeds; R: Liquid feed of 10% fermented tamarind seeds; R2:
Liquid feed of 20% fermented tamarind seeds; R3: Liquid feed of 30% fermented tamarind seeds.
Nutrient Content on Liquid Feed with Different Percentages of Tamarind Seeds Based on Table 3, it is known that the results of the analysis of variance showed that the use of different percentages of tamarind seeds in the feed had a very significant effect (P<0.01) on the nutrient content of the fermented liquid feed. This indicates that the higher the percentage of using tamarind seeds in the feed, the water content decreases due to imbibition into the cell contents so that the availability of nutrients is higher.
Duncan's test results showed that there was no significant difference (P>0.05) in dry matter content between liquid feed without the use of tamarind seeds and the use of 10%
and 20% tamarind seeds. However, it was significantly different (P<0.05) with liquid feed containing 30% tamarind seeds in the feed. In addition, it can be seen that the higher the use of tamarind seeds in liquid feed, the higher the dry matter content, due to the increased use of tamarind seeds, the more water is needed and absorbed in the imbibition process into the contents of the tamarind seed cells which causes the decomposition of nutrients. According to Narsih, Yunianta, and Harijono (2008), immersion causes biological changes, namely the breakdown of various complex components into simpler compounds.
Crude protein content based on Duncan's test results showed that there was a significant difference (P<0.05) between liquid feed without the use of tamarind seeds
and the use of 10% and 30% tamarind seeds, but not significantly different (P>0.05) with the use of tamarind seeds. 20% acid in the feed, and an increase in crude protein content with the addition of the percentage of tamarind seeds in the feed. The increase in crude protein content was in line with the increase in dry matter. This happens because the water imbibition process causes the expansion of the cell contents and causes the seed coat to expand and soften.
This condition makes it easier for microorganisms that develop in the fermentation process, namely LAB to take advantage of nutrients and produce enzymes that function to break down complex nutrients such as proteins into simple ones.
Tandrianto, Mintoko, and Gunawan (2014) stated during fermentation lactic acid bacteria Lactobacillus plantarum produces protease enzymes present in the fermentation process so that the dissolved protein levels increase.
This fact also occurs in the crude fiber content which aligned with the increase in the percentage of tamarind seeds in the liquid feed. The increase in crude fiber was because the tamarind seeds used were whole tamarind seeds without removing the peel where the availability of crude fiber and tannins was mostly located in the tamarind seed coat as a plant cell wall forming. This increase in crude fiber content indicates an increase in the fiber fraction in this case lignin Hernaman et al (2017) as a plant cell wall constituent. Furthermore, it was stated
DOI: 10.21776/ub.jiip.2023.033.02.06 192 that lignin was indigestible while crude fiber
had a fraction other than lignin, namely cellulose and hemicellulose which could be digested.
Duncan's test results showed that there was no significant difference (P>0.05) in the NFE content between liquid feed without the use of tamarind seeds and the use of 10%
tamarind seeds, but it was significantly different (P<0.05) with the use of 20% and 30% tamarind seeds. There was also an increase in NFE along with the use of tamarind seeds in the liquid feed. The increase occurred due to an increase in dry matter. It is stated so because NFE is part of carbohydrates which is also part of organic matter and dry matter.
In addition, the increase in tamarind seeds also increases the tannin content and will stimulate the formation of the tannase enzyme which will break down the tannins.
The decomposed tannins will also cause the release of complex bonds with carbohydrates and proteins so that they become more available. It is stated so because the presence of tannin which is a phenolic component can interact with proteins to form insoluble complex bonds Narsih, Yunianta, and Harijono (2018), as well as carbohydrates.
The results of Duncan's test of crude fat content in Table 3 show that there is no significant difference (P>0.05) in crude fat content between liquid feed without the use of tamarind seeds and the use of 10%
tamarind seeds, but it is significantly different (P<0.05) with the use of 20% and 30% tamarind seeds and a decrease in crude fat content.
This is because the increasing use of tamarind seeds in the liquid feed will also change the composition of other liquid feed ingredients (Table 2), in this case, the more use of tamarind seeds, the lower the use of bran and MBM which have a high crude fat content in the feed. That's because there is the dissolution of fat. The presence of immersion can activate lipase enzyme activity which can produce several short- chain free fatty acids that are easily soluble
in water (Pangastuti, Affandi, and Ishartani 2013). Thus, Mulia et al (2015) stated that the decrease in crude fat content in the fermentation process was due to the use of fat by microorganisms for their metabolic activities.
Based on Table 3, it can be seen that Duncan's test showed that there was no significant difference (P>0.05) in ash content between liquid feed without the use of tamarind seeds and 10% and 20%
tamarind seeds, but it was significantly different (P<0.05) with the use of tamarind seeds. 30% tamarind seeds in the feed. Table 3 also shows that there was an increase in ash content as the percentage of tamarind seeds used in liquid feed increased. The increase in ash content is due to an increase in the dry matter because ash or inorganic materials are part of the dry matter after being separated from the water.
Anti-nutrient (tannins) on Liquid Feed with Different Percentages of Tamarind Seeds
Fermentation of liquid feed made from tamarind seeds not only affects the nutritional content but also affects the anti- nutritive content of tannins. The results of Duncan's test showed that there was no significant difference (P>0.05) in anti- nutrient (tannins) content between liquid feed without the use of tamarind seeds (0%) and the use of 10% tamarind seeds, but it was significantly different (P<0.05) with the use of tamarind seeds. 20% and 30%. Table 3 also shows that there was an increase of 32.47% in the anti-nutrient (tannins) content with the use of 20% tamarind seeds and 35.89% with the use of 30% tamarind seeds.
The increase in anti-nutrient (tannins) content is due to the increasing use of tamarind seeds, the availability of crude fiber content, and tannins which are abundant in the tamarind seed coat also increases. Setiarto and Widhyastuti (2017) stated that lactic acid bacteria will produce tannase enzymes after 24 hours of growth in a medium containing 2 g of tannic acid in a medium without oxygen.
DOI: 10.21776/ub.jiip.2023.033.02.06 193 Content of Lactic Acid Bacteria (LAB) on
Liquid Feed with Different Percentages of Tamarind Seeds
The data in Table 3 shows that the population of lactic acid bacteria in the fermented liquid feed that uses tamarind seeds is up to 30% the same as the fermented liquid feed without the use of tamarind seeds. This indicates that sufficient nutrients are available in the fermented liquid feed substrate to be used in the activity of lactic acid bacteria.
This is because the fermentation that occurs is a liquid-feed fermentation that uses water. The presence of water in the feed causes the softening of the cell walls so the process of imbibition of water into the feed cells causes the feed material to expand and soften. This is the statement by Polhaupessy and Sinay (2014) that the imbibition process or the entry of water into the seeds causes the cells to swell. This causes the
microorganisms that develop in the fermentation process to make it easier to use the available nutrients for life. Lumowa and Nurani (2014) state that during the immersion process, spontaneous fermentation will occur which causes lactic acid bacteria (LAB) to develop and will utilize available nutrients.
Fiber fraction content a fermented liquid feed with different percentages of tamarind seeds
Based on the data in Table 4, it is known that the analysis of variance showed that the percentage of the use of tamarind seeds in the liquid feed had a very significant effect (P<0.01) on the content of fiber fractions (NDF, ADF, Cellulose, and lignin) and had a significant effect (P<0.05) on the hemicellulose content. Generally, the fiber fraction content increases with the increase in the percentage of use of tamarind seeds in the liquid feed.
Table 4. Fiber fraction content a fermented liquid feed with different percentages of tamarind seeds
Treatment Fiber Fraction (%)
NDF ADF Hemicellulose Cellulose Lignin
R0 34.42±2.44b 13.85±0.94d 20.52±1.48a 10.52±0.75c 2.59±0.30d R1 34,52±1.14b 15.76±0.76c 18.35±0.36b 11.41±0.68b 3.53±0.21c R2 38.68±2.46a 19.62±1.04b 19.14±1.60ab 12.12±0.45b 7.14±0.56b R3 40.91±2.10a 21.68±0.42a 19.28±1.64ab 12.85±0.42a 8.74±0.37a Note: a, b, c, d Means in the same column with different superscripts differ significantly (p<0.05); R0: Fermented liquid feed of 0% tamarind seeds; R1: Liquid feed of 10% fermented tamarind seeds; R2: Liquid feed of 20%
fermented tamarind seeds; R3: Liquid feed of 30% fermented tamarind seeds
Duncan's test showed that there was a significant difference (P<0.05) in NDF content between liquid feed without the use of tamarind seeds and the use of 10%
tamarind seeds with the use of 20% and 30%
tamarind seeds and there was no significant difference (P>0.05) between the percentage use of tamarind seeds of 20% and 30%.
Increasing the percentage of tamarind seeds causes more tamarind seeds to be available in the fermented liquid feed. The presence of these tamarind seeds increases the amount of fiber which will affect the fiber fraction.
Besides that, in the fermentation process, microorganisms are also active, especially from the bacteria group. The increase in the
number of these bacteria also has an impact on the amount of fiber contained in the ration.
The same thing was stated by Fariani and Akhadiarto (2012) that the increase in crude fiber content (such as NDF) was caused because the organic matter that was easily digested had been broken down a lot by lactic acid bacteria during the ensilage process so that what remained was organic material that was not easily digested such as crude fiber.
The ADF content in Table 4 showed an increase with the increase in the use of tamarind seeds in liquid feed fermentation and Duncan's test showed that there was a
DOI: 10.21776/ub.jiip.2023.033.02.06 194 significant difference (P<0.05) in ADF
content between each liquid feed treatment.
This is also because the feed ingredients are submerged in water so that they become softer while breaking complex bonds.
Lumowa and Nurani (2014) stated that during immersion there will be spontaneous fermentation which causes lactic acid bacteria to develop.
Duncan's test results showed that there was a significant difference (P<0.05) in hemicellulose content between liquid feed without using tamarind seeds and liquid feed using 10% tamarind seeds, but there was no significant difference (P>0.05) with liquid feed using tamarind seeds. tamarind seeds 20% and 30%. Likewise, there was no significant difference (P>0.06) between liquid feed containing 10% tamarind seeds with 20% and 30% in the fermented liquid feed. This shows that microorganisms can digest hemicellulose well even though the percentage is increasing. Wibisono et al (2011) stated that in the cell wall, lignin together with hemicellulose forms a matrix (cement) that binds the fine cellulose fibers.
Duncan's test of cellulose content showed significant differences (P<0.05) between fermented liquid feed without using tamarind seeds using 10%, 20%, and 30%
tamarind seeds. Likewise, fermented liquid feed containing 30% tamarind seeds was significantly different (P<0.05) with liquid feed without tamarind seeds and using 10%
and 20% tamarind seeds, while there was no significant difference (P>0.05) between liquid feed using tamarind seeds 10% by 20%. This shows that the higher the use of tamarind seeds, the more tannase enzymes are produced to break down tannins so that the fiber fraction becomes more available.
This is because the ability to degrade tannins is very different between bacteria, fungi, and fungi (Aguilar-Z'arate et al 2014).
Like the ADF content, the lignin content also had a significant difference (P<0.05) between each liquid feed treatment. The presence of lactic acid bacteria (LAB) in the fermentation process will stimulate the production of tannase
enzymes which will degrade tannins so that the complex bonds between tannins with proteins and carbohydrates can be broken and nutrients become more available.
According to Martina et al (2015), naturally, lignin is difficult to degrade and only a few microorganisms can degrade it.
CONCLUSION
The use of tamarind seeds up to 30%
in liquid feed increases nutrient content (dry matter, crude protein, crude fiber, nitrogen- free extract (NFE), and ash), tannins, and fiber fraction content (NDF, ADF, cellulose, hemicellulose, and lignin), but was able to reduce the crude fat content and had no impact on the population of lactic acid bacteria.
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