Research Article

Fermentation of Corn Tumpi with Bacillus Amyloliquefaciens: Optimization of Fermentation Duration and Inoculum Dosage for Superior Poultry Feed

Romi Andika1*, Wizna2, Mirzah2, Yetti Marlida2

1Doctoral Program in Animal Science, Faculty of Animal Science, Andalas University, Padang, Indonesia; 2Faculty of Animal Science, Andalas University, Padang, Indonesia.

Received | December 29, 2024; Accepted | February 01, 2025; Published | February 22, 2025

*Correspondence | Romi Andika, Doctoral Program in Animal Science, Faculty of Animal Science, Andalas University, Padang, Indonesia; Email: [email protected]

Citation | Andika R, Wizna, Mirzah, Marlida Y (2025). Fermentation of corn tumpi with Bacillus amyloliquefaciens: Optimization of fermentation duration and inoculum dosage for superior poultry feed. J. Anim. Health Prod. 13(1): 160-165.

DOI | https://dx.doi.org/10.17582/journal.jahp/2025/13.1.160.165

ISSN (Online) | 2308-2801

Copyright © 2025 Kumar et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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

The success of poultry farming is influenced by several factors, with the quality and quantity of feed being the primary determinants of the industry’s development. According to Maulana et al. (2021), feed is a critical factor in poultry farming because it accounts for 70–80% of production costs and directly influences livestock performance. Poultry feed is categorized based on nutritional content, with energy feed holding the largest proportion, accounting for 45–55%. Corn is one of the most widely used energy feed sources globally in poultry farming (Hidayat et al., 2024).

The use of corn as an energy feed source for poultry remains irreplaceable due to its quality and high nutrient digestibility, which maximizes poultry performance. However, competition for corn usage in feed impacts its availability, encouraging innovations in developing alternative feeds derived from corn by-products, such as corn tumpi (Yanuarinto et al., 2023).

Corn tumpi are a by-product of corn kernel processing and hold great potential as poultry feed ingredients. The nutritional composition of corn tumpi includes: water content 11.54%, dry matter 88.46%, crude protein 5.04%, crude fat 1.60%, crude fiber 30.93%, and ash 7.67% (Pratiwi and Basri, 2020).

Improving the nutritional quality of feed requires an innovative processing technology approach to ensure that the nutritional needs of animals are met efficiently and sustainably. One of the technologies commonly used is fermentation, which involves microorganisms such as bacteria, yeast, or fungi (Zumiarti et al., 2024). The fermentation process allows the transformation of raw feed materials into products with higher nutritional value by breaking down complex compounds such as fiber, lignin, or anti-nutrients into more digestible substances, such as short-chain fatty acids and soluble proteins (Maulana et al., 2024). Additionally, fermentation can enhance the availability of vitamins, minerals, and other bioactive compounds that support the health and performance of livestock.

Fermentation is an effective method for enhancing the nutritional value and digestibility of feed ingredients. It has the potential to improve nutrient content (Anyiam et al., 2023). Bacillus amyloliquefaciens is a probiotic bacterium capable of producing enzymes that degrade crude fiber, increase protein levels, and improve the amino acid profile of feed ingredients. This bacterium produces enzymes such as amylase (Gangadharan et al., 2011), cellulase, protease, and xylanase (Zhao et al., 2022). The key factors influencing the success of fermentation are fermentation duration (Sugiarti et al., 2024) and inoculum dosage (Maulana et al., 2024). This study was conducted to determine the optimal fermentation duration and inoculum dosage for fermenting corn tumpi with Bacillus amyloliquefaciens. The objective was to identify the best fermentation parameters as an innovation for superior poultry feed development.

MATERIALS AND METHODS

Fermentation of Corn Tumpi with Bacillus Amyloliquefaciens

A 100-gram sample of corn husks was taken and sterilized in an autoclave for 15 minutes at 121℃ and 1 atm pressure. After sterilization, the medium was allowed to cool to room temperature. The inoculum of Bacillus amyloliquefaciens was added at the respective treatment dosages (1%, 3%, and 5%), mixed thoroughly until homogenized, and incubated for the designated fermentation durations (2, 4, and 6 days). After the incubation process was completed, the medium was dried in an oven at 60℃ until fully dried to obtain the fermented product (Zumiarti et al., 2024).

Experimental Design

This experiment used a 3x3 factorial design applied in a completely randomized design (CRD), with three replications per treatment to ensure more accurate and reliable results. The 3x3 factorial design means that two factors are tested, each with three levels of treatment. The first factor, fermentation duration, has three different levels: 2 days, 4 days, and 6 days. This is intended to observe how the length of fermentation time affects the desired outcomes. The second factor, inoculum dosage, also has three different levels: 1%, 3%, and 5%. This inoculum is used to initiate the fermentation process, and varying inoculum dosages will help determine the impact of microorganism concentration on the fermentation process and the quality of the final product. By using this design, the experiment aims to analyze the interactions between these two factors and their individual effects on the fermentation results.

Parameters

The measured parameters included water content, dry matter, crude protein, crude fiber, crude fat, and ash content that were analyzed according to procedures of AOAC, (2016).

Data Analysis

All collected data were statistically analyzed using analysis of variance (ANOVA). Differences between treatments were determined using Duncan’s Multiple Range Test (DMRT) (Steel et al., 1997).

 

Table 1: Dry matter content of fermented corn tumpi with Bacillus amyloliquefaciens.

Fermentation Duration (Factor A)	Inoculum Dose (Factor B)			AverageNS
	B1 (1%)	B2 (3%)	B3 (5%)
A1 (2 days)	90.24	88.98	87.85	89.03
A2 (4 days)	90.06	88.02	87.86	88.65
A3 (6 days)	88.93	87.68	87.66	88.09
Average**	89.74a	88.23ab	87.79b
SEM				0.60

 

**Different superscripts in a row indicate very significant effects (P<0.01). NS: Non significant (P>0.05); SEM: Standard error of the mean.

 

RESULTS AND DISCUSSION

Dry Matter

Table 1, shows that the dry matter content of corn tumpi fermented with Bacillus amyloliquefaciens ranges from 87.66% to 90.24%. This study showed that there was no interaction (P > 0.05) between factor A (fermentation duration) and factor B (inoculum dosage), but factor B (inoculum dosage) had a highly significant effect (P < 0.01) on the dry matter content of corn tumpi.

Dry matter content with different fermentation durations and inoculum doses showed similar results. This is because all treatments involved fermentation, during which simple sugars were converted into gas and organic acids, leading to a decrease in the nutritional content of the corn tumpi. Dry matter is the portion of feed that excludes all water content (Amelia et al., 2022). The reduction in dry matter occurred because of the fermentation process, which produces gas and organic acids. Feed materials with high water-soluble carbohydrates undergo fermentation more rapidly (Marlida et al., 2023). The decrease in dry matter content is also due to the fermentation of sugars into lactic acid by microorganisms (Pratiwi and Basri, 2020).

 

Table 2: Water content of fermented corn tumpi with Bacillus amyloliquefaciens.

Fermentation Duration (Factor A)	Inoculum Dose (Factor B)			AverageNS
	B1 (1%)	B2 (3%)	B3 (5%)
A1 (2 days)	9.76	11.02	12.15	10.97
A2 (4 days)	9.94	11.98	12.14	11.35
A3 (6 days)	11.07	12.32	12.34	11.91
Average**	10.26b	11.77ab	12.21a
SEM				0.60

 

**Different superscripts in a row indicate very significant effects (P<0.01). NS: Non significant (P>0.05); SEM: Standard error of the mean.

 

Water Content

Table 2, illustrates that the water content of corn tumpi fermented with Bacillus amyloliquefaciens ranged from 9.76% to 12.34%. This study showed that there was no interaction (P > 0.05) between factor A (fermentation duration) and factor B (inoculum dosage), but factor B (inoculum dosage) had a highly significant effect (P < 0.01) on the water content of the corn tumpi. The changes in water content in this study showed similar results. This is because the dry matter content before fermentation was the same, and fermentation occurred in all treatments (Rahmatullah et al., 2020). During the fermentation process, bacteria convert substrates into simple products, and this process produces by-products such as water and carbon dioxide (Maulana et al., 2021).

Crude Protein

Table 3, demonstrates that the crude protein content of corn tumpi fermented with Bacillus amyloliquefaciens ranged from 7.47% to 10.59%. This study showed a highly significant interaction (P < 0.01) between factor A (fermentation duration) and factor B (inoculum dosage).

The high crude protein content in treatments A2B2 and A3B2 can be attributed to the higher inoculum doses (3% and 5%), which promoted abundant and even microbial growth, resulting higher microbial biomass and increased crude protein derived from microbial protein sources (MPS). This is consistent with the findings of Nuraini et al. (2015), where increasing the inoculum dosage led to more microbial growth and greater breakdown of materials.

 

Table 3: Crude Protein Content of Fermented Corn Tumpi with Bacillus amyloliquefaciens

Fermentation Duration (Factor A)	Inoculum Dose (Factor B)			Average**
	B1 (1%)	B2 (3%)	B3 (5%)
A1 (2 days)	7.75de	8.72cd	9.78abc	8.75A
A2 (4 days)	7.47e	10.59a	10.2ab	9.42AB
A3 (6 days)	9.66abc	10.45a	9.26bc	9.79A
Average**	8.29B	9.92A	9.75A
SEM				0.33

 

a-e: Different lowercase superscripts in a row or column indicate significant difference between treatments (P<0.05). **Different uppercase superscripts in a row or column indicate very significant difference between averages (P<0.01). SEM: Standard error of the mean.

 

The lower protein content in treatment A3B3 (9.26%) was observed because of the fermentation duration and the high inoculum dosage, which depleted the nutrients in the fermentation substrate as the microbes entered the death phase. As a result, the available nutrients were exhausted. The higher the inoculum dosage, the faster microbial development during fermentation; however, the available nutrients were consumed quickly (Maulana et al., 2024).

Crude Fat

Table 4, show that the crude fat content of corn tumpi fermented with Bacillus amyloliquefaciens ranged from 1.55% to 1.90%. This study showed no interaction (P > 0.05) between factor A (fermentation duration) and factor B (inoculum dosage), but factor B (inoculum dosage) had a highly significant effect (P < 0.01) on the crude fat content of the corn tumpi.

 

Table 4: Crude fat content of fermented corn tumpi with Bacillus amyloliquefaciens.

Fermentation Duration(Factor A)	Inoculum Dose (Factor B)			AverageNS
	B1 (1%)	B2 (3%)	B3 (5%)
A1 (2 days)	1.90	1.68	1.64	1.74
A2 (4 days)	1.82	1.59	1.77	1.73
A3 (6 days)	1.83	1.55	1.83	1.73
Average**	1.85a	1.61b	1.75ab
SEM				0.06

 

**Different superscripts in a row indicate very significant effects (P<0.01). NS: Non significant (P>0.05); SEM: Standard error of the mean.

 

The decrease in crude fat in the treatment B2 is due to the lipase enzyme produced by Bacillus amyloliquefaciens, which breaks down the crude fat in the corn tumpi into simpler products. Microorganisms then use these products as an energy source. Certain microorganisms can produce lipase to catalyze the breakdown of triglycerides into free fatty acids and glycerol. Microorganisms use the breakdown products of triglycerides as an energy source for growth and development during fermentation (Munandar et al., 2024).

Crude fat in feed consists of triglycerides, molecules formed from one glycerol and three fatty acids. Fat provides the highest energy content compared to protein or carbohydrates (9 calories per gram). Lipase can break down fat into simpler forms, reducing the complex fat content in the substrate.

Crude Fiber

Table 5, show that the crude fiber content of fermented corn tumpi with Bacillus amyloliquefaciens ranges from 10.53% to 13.57%. This study shows a significant interaction (P<0.05) between factor A (fermentation duration) and factor B (inoculum dosage). Both factor A (fermentation duration) and factor B (inoculum dosage) had highly significant effects (P < 0.01) on the crude protein content of the corn tumpi. Factor A (fermentation duration) shows a significant effect (P<0.05), while factor B (inoculum dosage) shows a highly significant effect (P<0.01) on the crude fiber content of corn tumpi.

 

Table 5: Crude Fiber Content of Fermented Corn Tumpi with Bacillus amyloliquefaciens.

Fermentation Duration (Factor A)	Inoculum Dose (Factor B)			Average*
	B1 (1%)	B2 (3%)	B3 (5%)
A1 (2 days)	13.01ab	12.98ab	10.56c	12.18A
A2 (4 days)	13.57a	10.84c	10.76c	11.72AB
A3 (6 days)	11.68bc	10.64c	10.53c	10.95B
Average**	12.75A	11.48A	10.62B
SEM				0.50

 

a-e Different lowercase superscripts in a row or column indicate significant difference between treatments (P<0.05).**Different uppercase superscripts in a row or column indicate very significant difference between averages (P<0.01). SEM: Standard error of the mean.

 

The best crude fiber content in this study was found in treatments A2B2, A3B2, A2B3, and A3B3, due to the longer fermentation times (4 and 6 days), which provided optimal conditions for Bacillus amyloliquefaciens to grow and produce cellulase enzymes, thereby reducing the crude fiber content. Nuraini et al. (2019) reported that fermentation duration influences cellulase enzyme activity, which significantly impacts the reduction of crude fiber content.

The lowest crude fiber content was found in treatment A1B3 (10.56%). This was due to the highest inoculum dosage (5%), which accelerated the growth of microorganism biomass during fermentation. Maulana et al. (2024) reported that inoculum dosage influences the biomass of microorganisms in the fermentation substrate, which increases cellulase enzyme activity. This enzyme helps degrade crude fiber, leading to a reduction in its content in the fermented product.

Crude fiber is a complex carbohydrate that poultry cannot digest efficiently because they do not produce cellulase enzymes. This enzyme is produced only by microorganisms or single-celled organisms. Crude fiber consists of cellulose, hemicellulose, and lignin. In large quantities, crude fiber reduces nutrient digestibility, but small amounts can benefit poultry by aiding in intestinal movement (Yanuarianto et al., 2024).

The breakdown of crude fiber by microorganisms during fermentation results in: firstly sellulase enzymes hydrolyzing cellulose in crude fiber into cellobiose, which then converts into glucose; Secondly hemicellulase enzymes breaking down hemicellulose into simple sugars like xylose and arabinose; and Finallly ligninase enzymes breaking the aromatic bonds in lignin, which protects cellulose and hemicellulose from enzymatic access (Cahyono et al., 2024).

Ash

In Table 6, the ash content of fermented corn husk with Bacillus amyloliquefaciens ranges from 4.35% to 5.64%. This study shows that there is no interaction (P>0.05) between factor A (fermentation duration) and factor B (inoculum dosage). Factor A (fermentation duration) shows a significant effect (P<0.05), while factor B (inoculum dosage) shows a very significant effect (P<0.01) on the ash content of the fermented corn husk.

 

Table 6: Ash content of corn bran fermented with Bacillus amyloliquefaciens.

Fermentation Duration (Factor A)	Inoculum Dose (Factor B)			AverageNS
	B1 (1%)	B2 (3%)	B3 (5%)
A1 (2 days)	5.00	4.35	5.64	5.00
A2 (4 days)	5.15	5.25	5.44	5.28
A3 (6 days)	5.03	4.88	5.49	5.13
AverageNS	5.06	4.83	5.53
SEM				0.36

 

NS: Non significant (P>0.05); SEM: Standard error of the mean.

 

The ash content of the fermented corn husk with Bacillus amyloliquefaciens remains unaffected during fermentation. This is because, during the fermentation process, microorganisms do not produce inorganic substances but only utilize the organic components of the fermentation substrate. Nuraini et al. (2019) reported that microorganisms growing during fermentation utilize organic substances such as complex carbohydrates and proteins to grow, while inorganic substances are not affected by microbial activity during fermentation. The phytase enzyme produced by certain microorganisms can enhance the absorption of phosphorus minerals (Yanuartono et al., 2016). Changes in mineral content during fermentation occur due to physical processes such as leaching and dissolution in the fermentation liquid or the external addition of mineral substances (Ignacio et al., 2023).

CONCLUSIONS AND RECOMMENDATIONS

This study concludes that the optimal treatment for corn husk fermentation with Bacillus amyloliquefaciens was achieved with a fermentation duration of 4 days and an inoculum dosage of 3%. The nutritional composition of the fermented corn husk with Bacillus amyloliquefaciens is as follows: dry matter 88.02%, water content 11.98%, crude protein 10.59%, crude fat 1.59%, crude fiber 10.84%, and ash 5.25%.

ACKNOWLEDGMENTS

This research is supported by the Doctoral Dissertation Research Scheme based on the decree number 0459/E5/PG.02.00/2024 and the agreement/contract number 041/E5/PG.02.00.PL/2024. This research would not have been possible without the participation of the students and the technical assistance from the Feed Laboratory, Feed Industry Technology and Ruminant Nutrition Laboratory, Faculty of Animal Science, Andalas University, Indonesia.

NOVELTY STATEMENTS

This study provides novel insights into optimizing the fermentation process of corn tumpi using Bacillus amyloliquefaciens by determining the ideal combination of fermentation duration and inoculum dosage. The significant interaction between these factors on crude protein and crude fiber content highlights the potential of controlled fermentation to enhance the nutritional quality of corn tumpi. The findings contribute valuable knowledge to the field of feed technology by establishing an optimal fermentation condition (4 days fermentation, 3% inoculum dosage) that improves nutrient composition, making it a viable strategy for enhancing livestock feed efficiency.

AUTHOR’S CONTRIBUTIONS

Wizna designed the research, wrote the manuscript, and interpreted the data. Romi Andika was involved in data collection and contributed to manuscript preparation. Mirzah and yetti participated in the preparation and critical review of the manuscript.

Conflict of Interest

The authors declare that there is no conflict of interest related to this research, whether financial, personal, professional or academic.

REFERENCES

Amelia F, Andriani Y, Haetami K (2022). Utilization of Plants as Fish Feed Ingredients: A Review, Ruaya Journal: J. Res. Stud. Fish. Marine Sci., 10(1): 23–29. https://doi.org/10.29406/jr.v10i1.3360

AOAC (2016). Official methods of analysis of the association of the official analytical chemists. 20th ed. Washington, D.C, USA.

Anyiama PN, Nwuke CP, Uhuo EN, Ije UE, Salvador EM, Mahumbi BM, Boyiako BH (2023). Effect of fermentation time on nutritional, antinutritional factors and in-vitro protein digestibility of macrotermes nigeriensis-cassava mahewu, Measurement: Food, 11(March): 100096. https://doi.org/10.1016/j.meafoo.2023.100096

Cahyono TD, Sukaryani S, Purwati CS (2024). Nutrient Content of Fermented MA-11 Corn Pulp with Different Incubation Periods, 12(2): 70–74. https://doi.org/10.30598/ajitt.2024.12.2.70-74

Gangadharan D, Nampoothiri KM, Pandey A (2011). α-amylase production by Bacillus amyloliquefaciens using agro wastes as feed stock, Food Technol. Biotechnol., 49(3): 336–340.

Hidayat R, Khusairi A, Zakhiya M, Maulana F, Fajri F, Febrina BP, Sandri D, Susalam MK (2024). Improvement of Nutrients in Food Waste and Organic Kitchen Waste from Islamic Boarding Schools in Tanah Laut Regency, Indones. J. Anim. Husbandry, 26(3):128–137. https://doi.org/10.25077/jpi.26.3.128-137.2024

Ignacio D, José B, Daniel T (2023). Mineral and nutrient content of corn stover silage with livestock excreta and by-products rich in carbohydrates. Abanico Veterinario E-Issn, 2448-6132.

Marlida Y, Harnentis H, Nur YS, Ardani LR (2023). New probiotics (Lactobacillus plantarum and Saccharomyces cerevisiae). supplemented to fermented rice straw-based rations on digestibility and rumen characteristics in vitro, J. Adv. Vet. Anim. Res., 10(1): 96–102. https://doi.org/10.5455/javar.2023.j657

Maulana F, Nuraini N, Mirzah M (2021). Content and Nutrient Quality of Palm Oil Waste Fermented with Lentinus edodes. Indones. J. Anim. Sci., 23(2): 174. https://doi.org/10.25077/jpi.23.2.174-182.2021

Maulana F, Fajri F, Febrina BP, Sandri D, Hidayat R (2024). Improving the nutritional quality of rice bran by fermentation using rumen fluid inoculum of male Bali cows with different fermentation times. Jurnal Peternakan, 21(2): 308–317. https://doi.org/10.24014/jupet.v21i2.30721

Munandar I, Amalyadi R, Dwitya NA (2024). Substitution of MOL as Biostarter EM4 for Improving the Nutrient Quality of Fermented Waste of Corn Tumpi and Pulp in Sumbawa Regency, 15(2): 280–286. https://doi.org/10.47687/jt.v15i2.725

Nuraini NA, Djulardi A, Trisna (2019). Palm Kernel Cake Fermented with Lentinus edodes in the Diet of Quail. Int. J. Poult. Sci., 18(8): 387–392. https://doi.org/10.3923/ijps.2019.387.392

Nuraini, Marlida Y, Mirzah, Disafitri R, Febrian R (2015). Increasing the nutrient quality of coffee seed waste using Phanerochaete chrysosporium as alternative feedstuff. Jurnal Peternakan, 17(2):143–150. https://doi.org/10.25077/jpi.17.143-150.2015

Oscar Y, Azhary N, Muhamad A, Dilaga SH, Dahlanuddin, Imran T (2024). The Nutrient Composition of Fermented Maize Stover with Different Fermentors. Jurnal Biologi Tropis, 24 (1): 352 – 358. http://dx.doi.org/10.29303/jbt.v24i1.6466

Pratiwi H, Basri H (2020). Evaluation of Nutrients in Corn Pulp Fermented with Various Bioactivators, Faperta Uniki Journal, 1(1): 17–22.

Rahmatullah R, Hasnudi, Mirwandhono E, Patriani P, Ginting N, Siregar GAW (2020). The effects of fermentation time and em4 dose on nutrient content of kepoks peel as animal feed, J. Phys. Conf. Ser., 1542(1). https://doi.org/10.1088/1742-6596/1542/1/012030

Steel RGD, Torrie JH, Dickey D (1997). Principles and procedures of statistics: a biometric approach.

Sugiarti, Tintin Rostini PW (2024). The Effect of Fermentation Duration on the Characteristics of Complete Ration Silage, Proceedings of the 3rd National Seminar on Animal Husbandry Scholars, Kediri, February 7: 26–31.

Yanuartono A, Nururrozi A, Indarjulianto S (2016). Fitat dan fitase: Dampak pada hewan ternak. Jurnal Ilmu-Ilmu Peternakan, 26(3): 59–78. https://doi.org/10.21776/ub.jiip.2016.026.03.09

Yanuarianto O, Supriadin D, Saidi MR, Putra RA, Burhan (2023). Utilization of fermented corn husks as an alternative feed availability in Keli Village, Woha District. Jurnal Gema Ngabdi, 5(1): 123–127. https://doi.org/10.29303/jgn.v5i1.324

Yanuarianto O (2024). Biologi Tropis Tropis Jurnal The Nutrient Composition of Fermented Maize Stover with Different Fermentors, (2011).

Zhao H, Zheng Z, Zhang M, Wang Y, Zhang M, Yang Z (2022). Fermentation optimization of rennet-producing Bacillus amyloliquefaciens GSBa-1 for high-density culture and its kinetic model, Food Sci. Technol. (Brazil),: 42: 1–9. https://doi.org/10.1590/fst.40122

Zurmiati T, Trisna A, Rusli RK, Dewi YL, Handayani UF, Wizna (2024). Performance Enhancement in Male Bayang Ducks after Oral Administration of the Probiotic Bacillus subtilis FNCC 0059. Anim. Health Prod., 12(1): 11–16. https://doi.org/10.17582/journal.jahp/2024/12.1.11.16