Research Article

Effect of Local Microorganisms from Bali Cattle Rumen Fluid and Palmyra Sap on Fiber Fractions and Nutrient Composition of Fermented Rice Bran

Tri Anggarini Yuniwati Foenay1, Theresia Nur Indah Koni1*, Agustinus Paga1, Oswilda Henyati1, Yosefus Frederikus da-Lopez2

1Department of Animal Science, Politeknik Pertanian Negeri Kupang, Jl. Prof. Dr. Herman Yohanes Lasiana Kupang P.O.Box. 1152, Kupang, Indonesia; 2Department of Dryland Agriculture Management, Politeknik Pertanian Negeri Kupang, Jl. Prof. Dr. Herman Yohanes Lasiana Kupang P.O.Box. 1152, Kupang, Indonesia.

Received | October 31, 2024; Accepted | February 03, 2025; Published | March 28, 2025

*Correspondence | Theresia Nur Indah Koni, Department of Animal Science, Politeknik Pertanian Negeri Kupang, Jl. Prof. Dr. Herman Yohanes Lasiana Kupang P.O.Box. 1152, Kupang, Indonesia; Email: Indahkoni@gmail.com

Citation | Foenay TAY, Koni TNI, Paga A, Henyati O, da-Lopez YF (2025). Effect of local microorganisms from bali cattle rumen fluid and palmyra sap on fiber fractions and nutrient composition of fermented rice bran. Adv. Anim. Vet. Sci. 13(4): 860-865.

DOI | https://dx.doi.org/10.17582/journal.aavs/2025/13.4.860.865

ISSN (Online) | 2307-8316; ISSN (Print) | 2309-3331

Copyright: 2025 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

INTRODUCTION

Rice bran, a by-product of rice milling and polishing (Bello et al., 2018; Munira et al., 2016), is a commonly used feed ingredient in livestock farming (Ahmad et al., 2019). Rice milling yields approximately 65% rice and 35% by-products, consisting of 23% husk and 10% bran and polishings (Superianto et al., 2018). The nutrient composition of rice bran includes 13-15% protein, 12-13% oil (Ahmad et al., 2019), metabolizable energy of 2980 kcal/kg, 11.4% crude fat, 0.07% calcium, 0.22% phosphorus, and 0.95% magnesium (Novita et al., 2017). However, it also contains 14-16% crude fiber (Ikhwanuddin et al., 2018), with crude fiber content in rice bran from Kupang reaching up to 24% (Koni et al., 2022). Fermentation effectively reduces crude fiber content (Bello et al., 2018; Nalar et al., 2014).

The fermentation process requires additives that accelerate fermentation, such as microbial inoculants (Wilkinson et al., 2003). Rumen fluid contains cellulolytic microorganisms capable of degrading crude fiber and fiber fractions (Ermalia et al., 2016; Nalar et al., 2014; Umbu et al., 2020). In addition to microbial inoculants, easily fermentable carbohydrate sources such as palmyra sap, sugar water, tapioca, or corn can act as fermentation substrates. Palmyra sap contains 4.0% fructose (Naiola, 2008), 36-78.86% sucrose, and 1.66-3.5% glucose (Humaidah et al., 2017; Vengaiah et al., 2017). Besides serving as a carbohydrate source, palmyra sap also provides microorganisms for fermentation, as it has been shown to contain Saccharomyces cerevisiae (Irmayuni et al., 2018) and lactic acid bacteria (Chayaningsih, 2006).

Rumen fluid and palmyra sap can be combined to formulate local microorganisms (MOL NR) following modifications from the study of Sio et al. (2022). MOL NR is a source of inoculant for microorganisms made from a mixture of Bali catle rumen fluid, palmyra sap, and granulated sugar water solution, which is incubated for three days. This MOL NR will be used as an inoculant for rice bran fermentation. The utilization of rice bran as a feed ingredient is limited by its high crude fiber and phytic acid content. Although MOL NR fermentation can address these issues, the optimal level of MOL NR for rice bran fermentation remains unknown. Koni et al. (2022) reported that the inclusion of 10% palmyra sap effectively reduced phytic acid in rice bran, while Ermalia et al. (2016) demonstrated that 30% rumen fluid inclusion, fermented for 72 hours, decreased fiber fractions in rice bran. Additionally, Namah et al. (2021) found that 30% rumen fluid inclusion reduced fiber fractions in banana peel, and Sio et al.(2022) showed that MOL NR derived from rumen fluid reduced fiber fractions in straw.

This study modifies the methodology of Sio et al. (2022) by preparing MOL NR NR from a mixture of Bali cattle rumen fluid (sourced from slaughterhouse waste), palmyra sap, and sugar water, fermented for three days. The MOL NR is used as an inoculant at varying inclusion levels during the fermentation of rice bran. Specifically, MOL NR is applied at 0%, 10%, 20%, and 30% (based on dry matter content) in rice bran fermented for six days. The objective of this study is to evaluate the effect of local microorganisms derived from a mixture of rumen fluid and palmyra sap (MOL NR) at different inclusion levels on the nutrient content and fiber fractions of fermented rice bran.

MATERIALS AND METHODS

Research Materials

The primary material used in this study was rice bran (Oryza sativa), purchased from a local distributor in Penfui, Kupang City. Palmyra sap was obtained from tappers in Lasina, Kupang, and Bali cattle rumen fluid was sourced from a local slaughterhouse. The equipment used included 20 plastic containers (1 kg capacity), bags, a digital Ohaus scale, trays, measuring cylinders, beakers, a pH meter, universal pH indicators, pipettes, a steamer, an autoclave, aluminum trays, a 60°C oven, heat-resistant plastic bags, and a blender.

Experimental Procedure

The experimental procedure is presented in the graphical format in Figure 1.

 

Preparation of Rice Bran

The rice bran was sieved to separate from husks and analyzed for dry matter (DM) content at the Nutrition and Animal Feed Laboratory, Politeknik Pertanian Negeri Kupang. The rice bran was sterilized using an autoclave at 121°C and 1 atm for 15 minutes. All equipment was washed with dish soap (Sunlight) and rinsed with antiseptic solution (20 mL Bayclin per 5 liters of water) before being air-dried. The plastic containers and their lids were sterilized by steaming for 10 minutes.

Production of Local Microorganism Inoculant (MOL NR)

The MOL NR inoculants, derived from Bali cattle rumen fluid, palmyra sap, and granulated sugar water solution, which is incubated for three days, was produced by following a modified protocol based on Sio et al. (2022). Rumen fluid was collected from the slaughterhouse and filtered to remove large particles. A mixture was prepared with 250 mL of rumen fluid, 250 mL of palmyra sap, and 250 mL of sugar water (200 g sugar dissolved in 50 mL water). This mixture was placed in a plastic container and incubated at room temperature for 7 days to develop the MOL NR inoculant.

Rice Bran Fermentation

The rice bran fermentation process followed the methodology described by Koni et al. (2022) with slight modifications. Rice bran (1 kg per replicate) was weighed according to the capacity of the containers. Each container was labeled with a unique number (1–20) to represent the experimental units, and treatments were assigned randomly using a lottery system.

The pH of the MOL NR inoculants were measured using a pH meter prior to its application. MOL NR was applied at four levels: 0%, 10%, 20%, and 30% of the rice bran dry matter. The inoculants were thoroughly mixed with the rice bran until homogeneous. The mixture was packed into the containers, compacted, covered with transparent plastic, and tightly sealed. Transparent tape was used to create anaerobic conditions. The containers were placed in a shaded area and fermented for six days following the protocol of Koni et al. (2022).

After six days, the containers were opened, and the fermented rice bran was inspected for mold. Any visible mold was removed, weighed, and discarded. The remaining rice bran was spread on labeled aluminum trays for physical evaluation (color, odor, and texture). A two grams sample from each replicate was taken twice for pH measurement.

The fermented rice bran was weighed, placed in paper envelopes, and sun-dried for two days. Once dry, it was ground, and 10% of the sample was set aside and labeled (1–20) for further analysis. These samples were sent to the laboratory for nutrient and fiber fraction analysis.

Nutrient and Fiber Fraction Analysis

The nutrient composition (dry matter, ash, crude fat, crude fiber, and crude protein) was analyzed following the procedures outlined by the Association of Official Analytical Chemists (AOAC, 2005). The fiber fractions, including neutral detergent fiber (NDF), acid detergent fiber (ADF), hemicellulose, cellulose, and lignin, were determined according to the method described by Van Soest et al. (1991).

Research Design

This study used a Completely Randomized Design (CRD) with four treatments and five replicates. The treatments were based on different levels of the local microorganism inoculants. (MOL NR) derived from cattle rumen fluid and palmyra sap, as follows: P0: Rice bran without MOL NR, P1: Rice bran with 10% MOL NR, P2: Rice bran with 20% MOL NR, P3: Rice bran with 30% MOL NR.

Research Parameters

The study evaluated the nutrient content of Dry Matter (DM), Crude Protein (CP), Crude Fiber (CF), Crude Fat (EE), and Ash. These parameters were measured using AOAC methods (AOAC, 2005). The fiber fractions analyzed included: Neutral Detergent Fiber (NDF), Acid Detergent Fiber (ADF), Hemicellulose, Cellulose, and Lignin, whilst fiber analysis was conducted according to the Van Soest method Van Soest et al. (1991).

Data Analysis

The nutrient content and fiber fractions were analyzed using analysis of variance (ANOVA). When significant effects (P < 0.05) were observed, Duncan’s New Multiple Range Test (DMRT) was employed to identify differences among the treatments.

RESULTS AND DISCUSSION

Effect of MOL NR Levels on Nutrient Content of Fermented Rice Bran

Fermentation with MOL NR significantly influenced the nutrient composition of rice bran, as detailed in Table 1. The process reduced dry matter, ash, and crude fiber content, while crude protein and crude fat levels showed minimal change (P > 0.05) across the different MOL NR concentrations.

Higher MOL NR levels led to a noticeable decrease in ash content. This can be attributed to the increased microbial activity during fermentation, where essential minerals are consumed to support their growth (Stanbury et al., 2003). The lowest ash levels were observed with 30% and 20% MOL NR, while unfermented rice bran had the highest ash content.

 

Table 1: Nutrient composition of fermented rice bran with different MOL NR doses.

Parameter	Treatment				P value
	P0	P1	P2	P3
Dry Matter (%)	61.4± 0.43c	60.9± 0.24bc	60.3± 0.58ab	60.1± 0.31a	0.003
Ash (%)	21.4± 0.43c	20.9± 0.24bc	20.3± 0.58ab	20.1± 0.31a	0.003
Crude Protein (%)	9.82± 0.24	10.2± 0.37	10.4± 0.62	10.3± 0.85	0.510
Crude Fiber (%)	27.6± 0.07b	27.1± 0.31ab	26.9± 0.35a	26.5± 0.48a	0.003
Crude Fat (%)	2.69± 0.35	2.52± 0.28	2.27± 0.23	2.50± 0.13	0.206

 

Note: a. b. ab. bc. c mean ± standard deviation followed by followed by different superscript letters within the same column indicate significant differences (P < 0.05). P0: Rice bran without MOL NR; P1: Rice bran with 10% MOL NR; P2: Rice bran with 20% MOL NR; P3: Rice bran with 30% MOL NR.

 

Although changes in crude protein were not statistically significant, fermentation improved protein levels compared to unfermented rice bran. Similar findings have been reported by Ibrahim and Usman, (2019) and (Ahmad et al., 2019), demonstrating that fermentation increases crude protein content.

A significant reduction in crude fiber content (P < 0.05) was observed with higher MOL NR levels. This reduction is likely due to cellulolytic microorganisms in MOL NR, which degrade fiber during fermentation. Previous studies, including those by Ermalia et al. (2016) and (Ahmad et al., 2019), also noted similar reductions, emphasizing the role of microbial enzymes such as cellulase and xylanase in breaking down fiber.

Interestingly, the crude fat content remained unaffected by MOL NR levels, likely because of limited lipase activity in the microorganisms present.Overall, using MOL NR in fermentation improved the nutritional quality of rice bran by reducing its ash and crude fiber content while enhancing its protein levels, making it a more valuable feed ingredient.

Effect of MOL NR Levels on the Fiber Fraction of Fermented Rice Bran

The fiber fraction content of rice bran treated with varying levels of MOL NR is summarized in Table 2. The addition of MOL NR significantly (P < 0.05) influenced the Neutral Detergent Fiber (NDF), Acid Detergent Fiber (ADF), hemicellulose, and cellulose content of rice bran.

NDF represents the structural components of plant cell walls, including cellulose, hemicellulose, lignin, silica, and fibrous proteins (Van Soest et al., 1991). High NDF levels in feed are less desirable for livestock as they reduce nutrient digestibility (Irawati et al., 2019). Fermentation with MOL NR reduced NDF content, likely due to the microbial breakdown of complex fiber fractions into simpler, more digestible compounds. Previous studies have also reported similar reductions in NDF content during fermentation. Koni et al. (2023) found that fermentation with palmyra sap reduced NDF content in rice bran by up to 56.57%. Likewise, Islam et al. (2022) observed a decrease from 49.61% to 31.31% after 12 hours of anaerobic fermentation using rumen fluid.

 

Table 2: Fiber fractions of fermented rice bran with different MOL NR doses

Parameters	Treatment				P value
	P0	P1	P2	P3
NDF (%)	76.5 ±4.63c	68.5 ±0.32b	58.3 ±1.06a	68.6 ±1.32b	0.000
ADF (%)	52.0 ±2.62c	49.9 ±0.51bc	47.5 ±0.54a	47.8 ±0.79ab	0.003
Hemicelulose (%)	24.6 ±2.22d	18.6 ±0.21b	11.4 ±0.67a	20.8 ±0.56c	0.000
Celulose (%)	21.5 ±1.85b	21.4 ±0.16b	17.5 ±1.59a	17.9 ±0.73a	0.001

 

Note: a. b. ab. c mean ± standard deviation followed by followed by different superscript letters within the same column indicate significant differences (P < 0.05). P0: Rice bran without MOL NR; P1: Rice bran with 10% MOL NR; P2: Rice bran with 20% MOL NR; P3: Rice bran with 30% MOL NR.

 

ADF, which includes cellulose, lignin, and silica, reflects the less digestible components of plant cell walls (Van Soest et al., 1991). While lignin is highly resistant to digestion, cellulose can be partially degraded. The decrease in ADF content observed with higher MOL NR levels is likely due to cellulolytic enzymes produced by MOL NR microorganisms. These enzymes, such as cellulase, degrade cellulose into simpler sugars like glucose, making it more accessible for digestion.

Hemicellulose, a digestible component of cell walls, is calculated as the difference between NDF and ADF. Fermentation with MOL NR altered hemicellulose levels, with the highest content observed in unfermented rice bran (P0). The microbial activity during fermentation likely contributed to changes in hemicellulose content, enhancing its digestibility and value as a feed component (Ati et al., 2020).

Cellulose, a primary component of plant cell walls, was also affected by MOL NR levels. Increasing the MOL NR concentration to 20% significantly reduced cellulose content, attributed to the action of cellulase enzymes produced by MOL NR microorganisms. Pamungkas (2011) noted that rumen fluid contains enzymes like cellulase, amylase, and protease, which play key roles in fiber degradation.

In conclusion, fermentation with MOL NR effectively improved the fiber fraction profile of rice bran by reducing NDF, ADF, and cellulose content, enhancing its digestibility and nutritional value as livestock feed.

CONCLUSIONS AND RECOMMENDATIONS

Increasing the dose of local microorganisms (MOL NR) derived from a mixture of rumen fluid and palmyra sap in the fermentation process resulted in a significant reduction in dry matter, ash content, crude fiber, and the fiber fractions of rice bran. Among the tested levels, the optimal MOL NR concentration for achieving these improvements was identified as 20% (P2), highlighting its effectiveness in enhancing the nutritional quality of rice bran through fermentation.

ACKNOWLEDGMENTS

The authors express gratitude to the Politeknik Pertanian Negeri Kupang for funding this research through the applied research scheme in 2024 under contract number 03/P3M/SP DIPA-023.18.2.677616/2024, dated March 27, 2024.

NOVELTY STATEMENTS

This study examines the use of local microorganisms derived from Bali cattle rumen fluid and palmyra sap and their effects on the nutritional quality of rice bran for animal feed.

AUTHOR’S CONTRIBUTIONS

Theresia Nur Indah Koni contributed to creating the research idea, designing experiments, analyzing data, and writing this article. Tri Anggarini Yuniwati Foenay and Agustinus Paga contributed data collection, Oswilda Henyati has contributed to carrying out proximate analysis and collecting research data, Yosefus Frederikus da-Lopez has helped with data analysis, and corrected the article.

Conflict of Interest

There was no conflict of interest in this research with other parties, both individuals and organizations.

REFERENCES

Ahmad A, Anjum AA, Rabbani M, Ashraf K, Awais MM, Nawaz M, Ahmad N, Asif A, Sana S (2019). Effect of fermented rice bran on growth performance and bioavailability of phosphorus in broiler chickens. Indian J. Anim. Res., 53:361–5.

AOAC (2005). Official methods of analysis of the association of analytical chemist. Association of Official Analytical Chemist, Inc, Virginia USA.

Ati S, Kleden MM, Yunus M (2020). Effect of length time of corn cob meal fermentation of using effective microoeganism-4 (EM4) on changing of ADF, NDF, Cellulose and Lignin component. J. Peternak. Lahan Kering, 2: 1162–1170.

Bello OA, Ayanda OI, Aworunse OS, Olukanmi BI (2018). A Comprehensive review on functional properties of fermented rice bran. pharmacogn. Rev. 12:218–224. https://doi.org/10.4103/phrev.phrev_11_18

Chayaningsih H (2006). Identification of Lactic Acid Bacteria From Palm Sap and Its Application in Reducing Salmonella typhimurium and Aspergillus flavus in Cocoa Beans. Sekolah Pasca Sarjana IPB, Bogor.

Ermalia AAU, Sjofjan O, Djunaidi IH (2016). Evaluation nutritients of rice bran second quality fermented using rumen fluid. Bul. Anim. Sci., 40: 113–123. https://doi.org/10.21059/buletinpeternak.v40i2.8700

Humaidah N, Widjaja T, Budisetyowati N, Amirah H (2017). Comparative study of microorganism effect on the optimisation of ethanol production from palmyra sap (Borassus flabellifer) using response surface methodology. Chem. Eng. Trans., 56: 1789–1794.

Ibrahim, Usman (2019). Efficiency of rations with the use of fermented rice brant in growth phase native chickens. Tolis Ilmiah; J. Penelit., 1: 124–129.

IIkhwanuddin M, Putra AN, Mustahal (2018). Utilization of rice bran fermentation with Aspergillus niger on feed raw material of Tilapia (Oreochromis niloticus). J. Perikan. dan Kelaut., 8: 79–87.

Irawati E, Purnamasari E, Arsyad F (2019). Physical and nutritional qualities of water hyacinth silage (Eichornia crassipes) with different fermentation lengths. J. Peternak. 16: 18. https://doi.org/10.24014/jupet.v16i1.3679

Irmayuni E, Nurmila, Sukainah A (2018). Effectiveness of nira lontar (Borassus flabellifer) as an ingredient for the development of apem cake doungh. J. Pendidik. Teknol. Pertan. 4: 170–183. https://doi.org/10.26858/jptp.v4i0.7122

Islam KMS, Elsabagh M, Lv R, Dang HL, Sugino T, Obitsu T (2022). Anaerobic fermentation of rice bran using rumen liquor for desirable chemical changes as animal feed. J. Adv. Vet. Anim. Res., 9: 728–735. https://doi.org/10.5455/javar.2022.i642

Koni TN., Foenay TAY, Vertigo S (2023). The use of urea and palmyra sap (Borassus flabellifer) on the characteristics and nutrient composition of fermented rice bran. Adv. Anim. Vet. Sci., 11: 624–629. https://doi.org/10.17582/journal.aavs/2023/11.4.624.629

Koni TNI, Foenay TAY, Jehemat A (2022). Fermentation characteristics and chemical composition of fermented rice bran with different levels of palmyra sap (Borassus flabellifer). Livest. Res. Rural Dev., 34: 88.

Munira S, Nafiu LO, Tasse AM (2016). Performance of cross bred native chickens fed feed substituted with fermented rice bran with different fermenters. J. Ilmu dan Teknol. Peternak. Trop., 3: 21–29. https://doi.org/10.33772/jitro.v3i2.1683

Naiola E (2008). Amylolitic microbes of nira and laru from Timor Island, East Nusa Tenggara. Biodiversitas 9: 165–168. https://doi.org/10.13057/biodiv/d090302

Nalar HP, Irawan B, Rahmatullah SN, Muhammad N, Kurniawan AK (2014). Utilization of rumen fluid in the fermentation process as an effort to improve the nutritional quality of rice bran for animal feed, in: Nat. Semin. Location Specif. Agric. Technol. Innov. Banjarbaru, 563–568.

Namah M, Wea R, Koni TNI (2021). Tannin, calcium (Ca), and phosphorus (P) content of banana peel flour fermented by goat rumen fluid. J. Ilmu dan Teknol. Peternak. Trop., 8: 51–56.

Novita N, Sofyatuddin K, Nurfadillah N (2017). The effect of fermented rice bran (Saccharomyces cerevisiae) on the growth of Rotifera (Brachionus plicatilis). J. Ilm. Mhs. Kelaut. dan Perikan. Unsyiah 2: 268–276.

Pamungkas W (2011). Fermentation technology, an alternative solution in effort to use local feed ingredients. Media Akuakultur 6: 43. https://doi.org/10.15578/ma.6.1.2011.43-48

Sio S, Bira GF, Batu MS, Pardosi L, Mau RJ, Klau MO, Hoar J (2022). Organoleptic quality and nutrition of rice straw silage utilizing local microorganisms of cattle rumen fluid at different inoculum levels. J. Adv. Vet. Res., 12: 36–41.

Stanbury PF, Whitaker A, Hall SJ (2003). Principles of Fermentaton Technology. Butterworth Heinemann. Burlington, Burlington.

Superianto S, Harahap AE, Ali A (2018). Nutrition value of cabbage vegetable waste silage with rice bran addition and different duration of fermentation. J. Sain Peternak. Indones., 13: 172–181.

Umbu CJT, Hilakore MA, Amalo D (2020). Effect of fermentantion time using goat rumen fluid on changes of putak quality. J. Peternak. Lahan Kering 2: 1022–1028.

Van Soest P, Robertson JB, Lewis BA (1991). Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J. Dairy Sci., 74: 3583–3597. https://doi.org/10.3168/jds.S0022-0302(91)78551-2

Vengaiah PC, Murthy GN, Sattiraju M, Maheswarappa HP (2017). Vale added food products from palmyrah palm (Borassus flabellifer L). J. Nutr. Heal. Sci., 4: 2–5. https://doi.org/10.15744/2393-9060.4.105

Wilkinson JM, Bolsen KK, Lin CJ (2003). History of silage. americam society of agronomy, crop science society of america, soill science society of america, Madison. https://doi.org/10.2134/agronmonogr42.c1