AAVS_13_10_2305-2312

INTRODUCTION

Bali cattle is one of Indonesia’s local cattle, which is excellent because it has a high reproductive ability (Sutarno and Setyawan, 2016), high carcass percentage, lean meat, and high adaptability to the environment. Despite these advantages, revealed that there had been a decrease in the genetic quality and productivity of Balinese cattle. One of the efforts to improve the performance of Bali cattle is to provide additional feed in the form of feed concentrates. Concentrates are feeds that are rich in protein sources and or energy sources and can contain feed complements and or feed additives. Giving concentrates with a crude protein content (PK) of 15-16% is very suitable for fattening cows in the tropics because in this area, during the dry season, forage productivity decreases so that concentrate feed is needed to meet their feed protein needs (Budiari et al. 2020). Giving concentrates based on DDGS to steers, is economically feasible and able to improve the growth performance of finishing cattle (Eun et al., 2009).

The use of legumes as one of the concentrate materials aims to meet the protein needs of livestock (Wati et al., 2020). Some types of legumes have a high crude protein content that can be used as concentrate material. Indigofera is one of the feed legumes that contain high crude protein (PK), namely 27.89%, crude fat or ether extract (EE) of 3.70%, and crude fiber (SK) of 14.96% (Akbarillah et al., 2008) total digestibility nutrient 69-72% (Antari et al., 2022) with dry matter digestibility ranging from 76% and protein digestibility ranging from 83% (Abdullah, 2010). Lamtoro leaves have a fairly high protein content, reaching 25-35% based on dry matter compared to other forages (Ghos and Bandyopadhyay, 2007). Gamal is a legume plant that can grow quickly in arid areas. The nutritional content of Gamal forage (G. sepium) is a protein content of 25.7%, crude fiber 13.3%, ash 8.4%, and BETN 4.0%. Several experiments have shown that giving concentrates containing legumes can increase the productivity of young male goats’ PE. This study aims to determine the quality and digestibility of concentrates made from several types of legumes and their effect on the performance of Balinese cows.

MATERIALS AND METHODS

Sample preparation and analysis

Indigofera sp. legumes (Indigofera), Leucaena leucocephala (lamtoro) and Gliricidia sepium (Gamal) aged 60 days were obtained from the Gowa Experimental Garden. Indigofera, lamtoro and fresh Gamal leaves are harvested and then dried in the sun for three days (28–37°C), then ventilated for 42 hours, and then ground until they are in the form of flour. Indigofera flour, lamtoro flour and Gamal flour are then mixed with other ingredients into concentrates according to the treatment (Table 1). Samples were tested for chemical composition, which included dry matter (DM), organic matter (OM), ash, crude protein (CP), ether extract (EE), crude fiber (CF) and nitrogen-free extract (NFE) by following the AOAC (2005) procedure. Samples were also specified for neutral detergent fibers (NDF) and detergent fiber acids (ADF) (Van Soest et al., 1991).

In vitro procedure

The in vitro procedure was performed according to (Tilley and Terry, 1963). The first step is that the cow’s rumen fluid is filtered using a sterile filter cloth to separate between the supernatant and the precipitate. In the supernatant section, a pH analysis of rumen fluid, NH3, and VFA is carried out. The rest, in the precipitate part to analyze the digestibility of dry matter and the digestibility of organic matter. A fermenter tube that had been filled with 500 mg of sample was added 10 ml of rumen fluid and 40 ml of Mc. Dougall’s solution. The fermenter tube is shaken by flowing CO2 gas for 30 seconds (pH 6.5-6.9) and covered with ventilated rubber. The tube was inserted into a water bath shaker with a temperature of 39 oC, fermented for 4 hours for a sample of the pH value of the rumen, NH3, total VFA, and 48 hours of fermentation for the digestibility of dry matter/digestibility of organic matter sample. The termination of the fermentation process is carried out by opening the lid of ventilated rubber and then dripping two drops of HgCl2. Furthermore, an analysis of the digestibility of dry matter and the digestibility of organic matter is carried out.

Table 1: The nutritional content of legume-based concentrates.

Parameter	Factor B	Factor A
	Level legum	Types of legumes
	Level legum	A1	A2	A3
Dry matter (%)	B1	86.94	88.20	87.80
	B2	85.75	88.76	88.14
	B3	87.40	87.49	85.12
Organic matter (%)	B1	83.82	85.27	87.80
	B2	80.46	88.76	88.14
	B3	87.80	88.43	85.12
Crude protein (%)	B1	14.11	14.26	13.34
	B2	14.86	14.37	13.10
	B3	14.31	14.11	14.20
Ether extract (%)	B1	3.79	3.41	3.36
	B2	3.92	2.86	3.99
	B3	2.10	3.45	3.33
ADF (%)	B1	42.14a	47.36b	45.00b
	B2	43.26a	43.46a	41.61a
	B3	38.79a	45.52b	41.60a
NDF (%)	B1	58.04ab	61.75b	60.55b
	B2	61.09b	56.63a	55.68a
	B3	53.74a	59.91ab	56.13a

Different superscript letters (a, b) within a row row/column indicate significant differences (P < 0.05). Factor A types of legumes: A1= concentrate made from Indigofera flour, A2= concentrate made from lamtoro flour and A3 = concentrate made from Gamal flour. Factor II level of legumes in concentrates (B) respectively: B1= level 0%, B2= level 5 % and B3= level 10%.

In vivo procedure

Cows are kept in individual pens and fed twice a day, namely in the morning at 07.00 WITA given elephant grass and concentrates as much as 2 kg/head/day and in the afternoon at 16.00 WITA given elephant grass. Drinking water is given ad libitum. Feeding is carried out for four months and is carried out weighing the rest of the feed and weighing body weight once a month. The complete stages of the research are presented in the visual (Box-and-Arrow) shown in Figure 1.

Statistical analysis

The experimental design used was a factorial design of a complete randomized design pattern. Factor one is the type of legumes (A) respectively: A1= concentrate made from Indigofera flour, A2= concentrate made from lamtoro flour and A3= concentrate made from Gamal flour. Factor two is the level of legumes in the concentrate (B) respectively: B1= level 0%, B2= level 5 % and B3= level 10%. Each combination of treatments was repeated four times so that there was 36 experimental livestock. When the ANOVA results for a certain parameter show a significant difference at p<0.05, the Duncan test was applied to compare different treatments. Statistical analysis was performed by using SPSS statistics software version 23.

RESULTS AND DISCUSSION

Table 2 shows the results of the proximate analysis of various types of legumes in concentrates with a significant effect (P<0.05) on dry matter and organic matter, but there is no significant difference in the level of legumes (P>0.05). Concentrates made from lamtoro flour (A2) have a higher dry matter content than concentrates made from Indigofera flour (A1) and Gamal flour (A3). The measurement of concentrate dry matter is to determine the quality and affect the shelf life of the concentrate. The type and level of legumes in the concentrate have no significant effect (P>0.05) on the crude protein content of the concentrate.

Table 2: Rumen fermentation.

Parameter	Level legum	Types of legumes			Average
Parameter	Level legum	A1	A2	A3
VFA (mMol)	B1	93.98	91.29	95.07	90.11a
	B2	118.86	102.09	107.38	109.44b
	B3	111.95	117.98	111.44	113.79b
Average		104.93	103.78	104.63
NH3 (mMol)	B1	8.47	8.52	8.01	8.33a
	B2	8.94	8.42	7.96	8.44a
	B3	10.11	9.89	9.34	9.44b
Average		9.17b	9.11b	8.43a
pH	B1	6.93 e	6.82abc	6.85bcd	6.87
	B2	6.80 ab	6.95e	6.90de	6.88
	B3	6.86 bcd	6.88cde	6.77 a	6.84
Average		6.86	6.89	6.84

Different superscript letters (a, b, c, d, e) within a row/column indicate significant differences (P < 0.05). Factor A types of legumes respectively: A1 = concentrate made from Indigofera flour, A2 = concentrate made from lamtoro flour and A3 = concentrate made from Gamal flour. Factor II level of legumes in concentrates (B) respectively: B1 = level 0%, B2 = level 5 % and B3 = level 10%.

Table 3: Feed consumption rates.

Parameter	Level legum	Types of legumes			Average
Parameter	Level legum	A1	A2	A3	Average
Total dry matter intake (g/d)	B1	6578.26b	6738.23b	3427.86a	5581.45
	B2	6741.44b	6714.71b	3592.50a	5682.88
	B3	6622.67b	6734.46b	6798.24b	6718.46
Average		6647.46	6729.13	4606.20
Total organic matter intake (g/d)	B1	5302.40b	5267.41b	2772.69a	4447
	B2	5424.40b	5519.87b	2912.28a	4618.85
	B3	5513.93b	5513.06b	5407.58b	5407.68
Average		5413.58	5433.45	3697.51
Crude protein intake (g/d)	B1	1040.85	1062.00	917.35	1006.73
	B2	1088.49	1022.96	961.21	1024.22
	B3	1039.50	1026.50	1033.05	1033.01
Average		1056.28b	1037.15b	970.53a

Different superscript letters (a, b) within a row indicate significant differences (P < 0.05). Factor A types of legumes respectively: A1= concentrate made from indigofera flour, A2= concentrate made from lamtoro flour and A3 = concentrate made from gamal flour. Factor II level of legumes in concentrates (B) respectively: B1= level 0%, B2= level 5 % and B3= level 10%.

Table 4: Influence of concentrates made from several types of legumes and their effect on the performance of Balinese cows.

Parameter	Level Legum	Types of legumes			Average
Parameter	Level Legum	A1	A2	A3	Average
Body weight, kg
Initial, kg	B1	100.75	99.50	94.00	98.08
	B2	100.12	102.50	101.25	101.29
	B3	94.25	85.00	98.50	92.58
Average		98.37	95.66	97.91
Final, kg	B1	122.00	116.50	108.50	115.66a
	B2	137.15	129.75	123.50	130.12b
	B3	126.37	109.25	122.25	119.29ab
Average		128.50	118.50	118.08
Average daily gain, kg/d	B1	0.24	0.23	0.16	0.19a
	B2	0.41	0.30	0.25	0.32b
	B3	0.36	0.36	0.26	0.29b
Average		0.33c	0.25b	0.22a
Feed conversion ratio	B1	28.23c	36.27c	22.05b	28.85
	B2	16.55a	22.20b	14.60a	17.78
	B3	17.03b	25.15c	25.89c	22.69
Average		20.61	27.87	20.84

Different superscript letters (a, b, c) within a row row/column indicate significant differences (P < 0.05). Factor A types of legumes: A1= concentrate made from Indigofera flour, A2= concentrate made from lamtoro flour and A3= concentrate made from Gamal flour. Factor II level of legumes in concentrates (B) respectively: B1= level 0%, B2= level 5 % and B3= level 10%.

The average nutrient consumption of treated feed during the study is listed in Table 3. The results of the various analysis showed that there was an interaction between the type of legumes and the level of legumes in the concentrate against the consumption of dry matter and organic matter.

The digestibility in the rumen of the nutrient feed treatment during the study is listed in Table 4. The results of the various analysis showed that the type and level of legumes in the concentrate had a significant effect (P<0.05) on dry matter and organic matter, and Duncan’s further test showed an interaction between the type of legumes and the level of legumes in the concentrate against the digestibility of dry matter and organic matter.

Table 5: Digestibility nutrient and nitrogen retention.

Parameter	Level legum	Types of legumes			Total	Average
Parameter	Level legum	A1	A2	A3	Total	Average
Dry matter (%)	B1	80.00b	80.63b	61.59a	888.92	74.07
	B2	80.23b	79.78b	62.69a	890.86	74.23
	B3	78.61b	79.46b	78.95b	948.12	79.01
Average		79.61	79.96	67.74
Organic matter (%)	B1	76.14b	75.78b	56.25a	208.12	69.39
	B2	75.43b	76.41b	57.67a	209.51	60.84
	B3	73.56b	76.68b	75.27b	225.51	75.17
Average		75.04	76.29	63.06
Crude protein (%)	B1	89.20b	90.86b	90.90b	271.83	90.61
	B2	91.20b	89.63b	89.93b	270.96	90.32
	B3	91.43b	90.46b	79.06a	267.54	89.18
Average		90.61	90.32	89.18
Nitrogen in feces (g/day)	B1	15.78e	12.92a	13.95b	42.85	14.22
	B2	15.70de	13.79b	15.11cd	44.60	14.87
	B3	12.86a	14.64c	15.31de	42.81	14.27
Average		14.78	13.78	14.79
Nitrogen in Urine (g/day)	B1	9.58a	10.89a	11.39a	31.86	10.62
	B2	15.39c	17.09c	17.51c	49.99	16.66
	B3	14.89b	15.66b	17.08b	47.63	15.88
Average		13.29	14.55	15.32
Nitrogen digested (g/day)	B1	132.36b	139.16b	141.76b	413.28	137.76
	B2	141.10b	137.80b	137.36b	416.25	138.75
	B3	144.46b	137.23b	60.86a	342.54	114.18
Average		139.31	138.16	113.33
Nitrogen retention (g/day)	B1	122.86	127.70	45.93	296.49	98.83
	B2	130.63	122.46	119.70	372.18	124.06
	B3	133.43	120.80	124.56	378.79	126.26
Average		128.97	123.65	96.73

Different superscript letters (a, b, c, d, e) within a row row/column indicate significant differences (P < 0.05). Factor A types of legumes: A1 = concentrate made from Indigofera flour, A2= concentrate made from lamtoro flour and A3= concentrate made from Gamal flour. Factor II level of legumes in concentrates (B) respectively: B1= level 0%, B2= level 5 % and B3= level 10%.

This study showed that the type of legumes in the concentrate had no significant effect (P>0.05) on VFA production, but the level of legumes in the concentrate had a real impact on VFA production (P<0.05). Table 5 shows that the higher the level of legumes in the concentrate, the higher the production of the resulting VFA.

However, the results of proximate tests show that the average crude protein content of A1 concentrate is higher than A2 and A3, which is 14.43%. The highest at the level of 10% at 14.21%; it is suspected that because Indigofera has a high protein content when mixed in the concentrate, it will increase the crude protein content of the concentrate. The protein value of Indigofera leaves varies from 25–28% and can even be up to 31%. Duncan’s multiple spacing test showed an interaction between legume type and legume level in the concentrate against ADF content. The lowest ADF content is A1 concentrate, while the highest is A2 concentrate. As with the ADF content, there is also an interaction between the type of legumes and the level of legumes in the concentrate to the NDF content. A3 concentrate has the lowest NDF content, while the highest NDF value is in A2 concentrate, and 10% has the lowest NDF content. The components of NDF and ADF are fractions of crude fibers /dry matter from feed (Fisher et al., 1985). Cell walls are chemically expressed in NDF (Neutral Detergent Fiber). NDF represents the content of the cell wall consisting of lignin, cellulose, hemicellulose and proteins that bind to the cell wall. Cell walls that are insoluble in acid detergents, namely ligno-cellulose, commonly called Acid Detergent Fiber (ADF). Chemically, A1 concentrate with a level of 10% is a concentrate that has better quality than other concentrates, because low NDF and ADF values indicate good feed quality.

The B3 treatment has the highest VFA value of 113.79 mM. Increased production of volatile fatty acids (VFA) may indicate the ease with which a nutrient in the feed, especially carbohydrates and proteins, is digested by rumen microbes (Izzatullah et al., 2018). Statistical tests of NH3 values showed a significant influence (P<0.05) on the type and level of legumes in the concentrate but did not show any interaction between the two treatments. The highest NH3 value at A1 (9.17 mM) was then A2 (9.11 mM), and the lowest was A3 (8.43 mM) as shown in Figure 2. At the legume level treatment, it shows that the higher the level of legumes in the concentrate, the higher the value of NH3. This result can occur because the concentrate made from Indigofera with a level of 10% has a high protein content and is easily degraded by the rumen. An increase in the concentration of NH3 in the rumen can occur because it is suspected that the feed ingredients contain crude protein, which is easily digested by rumen microbes (Sujarnoko et al., 2020). Although higher NH₃ values may indicate reduced nitrogen-use efficiency, they did not appear to compromise growth performance in this study.

The degree of acidity (pH) of the rumen fluid indicates fermentation process in the rumen. Treatment resulted in an average rumen pH of 6.8. pH values are within the normal range of rumen pH 5.5-7.5 (Franzolin et al., 2010), so that it does not interfere with rumen microbial activity, supports rumen microbial growth, and produces VFA and NH3. According to (Kamra, 2005), the optimum pH for the growth of rumen microbes is 6-6.9.

In general, it is said that changing the concentrate level in ruminant feed can significantly affect DMI and nutrient digestibility (Gizachew, 2012). Microbial manipulation with consentrate can improve feed digestibility and health status of Angus cow (Chen et al., 2021). Maximum feed consumption depends mainly on the balance of nutrients in digestion (Preston and Leng, 1984; Wilson and Kennedy, 1996). This is because nutritional needs are the main stimulant to convey the hypothesis of hunger. Furthermore, (Preston and Leng, 1984) stated that the imbalance of feed nutrients would affect feed consumption. The highest consumption of dry matter in A1B2 was 6741.44 g/d, and the highest consumption of organic matter at A1B3 was 5513.93 kg/d. The lowest amount of dry matter was at A3B1 (3427.86 g/d).

This shows that A1 concentrate at the level of 5%-10%s has a high level of palatability. According to Faverdin et al. (1995), palatability is an important factor that explains the differences in dry matter intake between feeds and among low-quality feed. Protein consumption showed a significant effect (P < 0.05) on the type of legumes and had no apparent effect (P > 0.05) on the level of legumes. The highest crude protein consumption was in the A1 treatment (1056.28 g/d), not significantly different from A2 (1037.15 g/d) and the lowest treatment, A3 (970.53 g/d). The high consumption of dry matter, organic matter and crude protein in A1 is due to the concentration having a lower ADF and NDF content than other concentrates. In the opinion of (Cherry, 1982), the higher fibre content in the ration causes the amount of ration consumption to decrease because the ration is “bulky” so that the ration consumed is limited.

The potential of feed to provide nutrients to livestock is determined through chemical analysis, but the actual value is indicated by the share lost after digestion, absorption and metabolism. Defined digestibility or digestibility is part of the feed nutrients that are not excreted in the faeces and are assumed to be the part absorbed by livestock. The highest digestibility of dry matter in the legume type is A2 (79.96 %), not much different from A1 (79.61%), at the highest legume level in B3, which is 79.01%, while the highest legume level is at 10% (79.01%).

This means that the quality concentrates containing legume lamtoro and Indigofera has a better digestibility of dry matter than Gamal concentrates, and concentrates containing 10% legumes have better quality than 0% and 5%. This is in accordance with the statement of (Chen et al., 2021) that the digestibility of DM in ruminants shows a high level of feed substances that can be digested by microbes and digestive enzymes in the rumen. The higher the percentage of DM digestibility of a feed ingredient, indicating that the higher the quality of the feed ingredient. As with the digestibility of DM, the average digestibility of real A2 and A1 organic matter is higher than that of A3, namely 76.29%, 75.04% and 63.06%, respectively. OM digestibility at the level of 10% (B3) is higher than 0% and 5%, namely 75.17%, 69.39% and 60.84%, respectively. Organic matter (OM) is a component of dry matter (DM); therefore, an increase in DM will also result in an increase in the OM content of the same material (Wahyuni et al., 2014). The results of the various analysis showed that the type and level of legumes in the concentrate had a significant effect (P<0.05), and there was an interaction with the digestibility of crude protein. Crude protein digestibility was highest in concentrates containing Indigofera legumes with a level of 10% (A1B3) of 91.43% and lowest in concentrates with Gamal legumes with 10% (A3B3) of 79.06%. This result is because concentrates containing Indigofera legumes have the highest crude protein content, while concentrates containing Gamal legumes have the lowest crude protein content compared to other concentrations. This is in line with what was stated by Sutrisno et al. (2013) that the high level of protein digestibility depends on the protein content of the feed ingredients and the amount of protein consumed. The values of N faeces and N Urine statistically show a noticeable difference (P<0.05), and there is an interaction between the type and level of legumes in the concentrate. The lowest faeces N value at A1B3 was 12.86 g/day, and the highest at A1B1 was 15.78 g/day. The amount of nitrogen in the faeces results from consumed N, which cannot be absorbed in the digestive tract and is disposed of as faeces. Increased N discharge through faeces will decrease the N absorbed or utilization of N by livestock. N that is less excreted through faeces is expected so that the utilization of N increases to support production, which means that the utilization of N rations is getting better (Tahuk et al., 2020). N urine values were lowest at A1B1 (9.58 g/day) and highest at A3B2 (17.51 g/day). Specifically, Indigofera concentrate had lower ADF and NDF contents compared to Leucaena and Gliricidia, which may have improved fiber digestibility and dry matter intake. In contrast, Leucaena is known to contain mimosine, and Gliricidia has secondary metabolites such as coumarins and tannins, which may negatively affect intake and nutrient utilization.

N-urine describes the amount of UN-used from N-available and N-absorbed through the gastrointestinal tract and with a slight mixture of N-endogenous amounts from the body itself. An increase in N urine leads to an increase in unused and N-absorbed. This means that the decrease in the value of profit or performance of livestock is due to an increase in the amount of N secreted in the form of urine (Sunarso, 2012). The undigested N value of the statistic shows a noticeable difference (P<0.05), and there is an interaction between the type and level of legumes in the concentrate. The highest digestible N value at A1B3 was 144.46 g/day, and the lowest at A3B3 was 60.86 g/day. Although nitrogen retention was not significantly different, the highest value was observed in A1 has the highest retention N value of 128.97 g/d, and A3 has the lowest retention N value of 96.73 g/d. These results illustrate that less N consumed is wasted through faeces and urine means that its digestibility and N retention increase. According to McDonald et al. (2012), the digestibility of nitrogen is obtained from the incoming nitrogen minus the nitrogen in the faeces.

The average statistical value of body weight gain of Balinese cattle given concentrates with different types and levels of legumes showed a real influence (P<0.05), but there was no interaction between the two. The Duncan test (Table 4 and Figure 3) showed that the average daily weight gain of A1 was higher than that of A2 and A3, namely 0.33 kg/h/day, 0.25 kg/h/day, 0.22 kg/h/day, respectively. At the legume level, B2 is higher but not different from B3 to the increase in body weight of Balinese cattle, namely 0.32 kg/h/day and 0.29 kg/h/day, respectively, while real B1 is the lowest in increasing the body weight of Balinese cattle by 0.19 kg/h/day. This result shows that giving concentrates containing Indigofera legumes can increase the body weight gain of Balinese cows better than concentrates with lamtoro and Gamal legumes. It is suspected to be because Indigofera has a high protein content, so when mixed in the concentrate, it will increase the crude protein content of the concentrate. This is in line with the opinion of Riswandi et al. (2017) that to improve the nutritional quality of feed is to supplement it using legumes. According to Muzzazinah (2016), the protein value of Indigofera leaves varies from 25–28% and can even be up to 31%. The results also showed that the addition of legumes to the concentrate at the level of 10-15% was able to increase the weight gain of cows compared to concentrates without legumes. Feed conversion ratio showed a noticeable difference (P<0.05), and there was an interaction between the type and level of legumes in the concentrate. The lowest ration conversion value was in the treatment with A3B2 (14.60) but did not differ markedly from A1B2 (16.55), and the ration conversion value was highest in the A2B1 treatment (36.2). These results show that feed containing concentrates with Gamal legumes and Indigofera is more efficient than concentrates with lamtoro legumes, and concentrates with a level of 10% l are more efficient than concentrates with 15% and 0% legumes.

CONCLUSION

Concentrates formulated with 10% Indigofera exhibited a high crude protein content, low ADF and NDF levels, elevated VFA and NH₃ concentrations, and an acidic rumen pH that promoted feed intake. These concentrates also demonstrated high crude protein digestibility and nitrogen utilization. Supplementation with 10% Indigofera concentrate improved weight gain in Bali cattle and enhanced feed efficiency.

ACKNOWLEDGEMENT

The authors are thankful to Indonesian Agency for Agricultural Research and Development, Ministry of Agricultural, Republic of Indonesia.

Novelty Statement

This study is novel as it compares three indigenous legumes—Indigofera, Leucaena, and Gliricidia—as concentrate ingredients for Balinese cattle, an approach that has not been thoroughly investigated previously. Moreover, the findings provide new evidence that incorporating 10% Indigofera flour into the concentrate significantly enhances crude protein digestibility and daily weight gain compared to the other legume sources.

AUTHOR’S CONTRIBUTION

NQ, UU: Format analysis.

NQ, AMP: Visualization.

NQ, AE, UU: Writing-review and editing.

AE: Conceptualization.

AE, AMP: Supervision.

AN, II: Data curation.

AN: Methodology.

NQ, AN, UU: Writing-original draft.

AN: Project-administration.

AMP: Resources.

II: Software and validation.

Ethical approval

The study procedures received approval from the Institutional Animal Care and Use Committee (Approval No. Balitbangtan/BPTP Sulsel/Rm/04/2020) and were implemented in accordance with recognized animal welfare guidelines, including provisions for housing, nutrition, and minimizing animal distress. The experiment was conducted at the Gowa Experimental Garden, South Sulawesi Province, Indonesia. This study used 36 female Balinese cows aged 1-1.5 years with an initial body weight ranging from ±85 - 100 kg.

Generative AI and AI-assisted technology statement

The authors used generative AI tools (e.g., ChatGPT, Grammarly) solely to assist with language editing and to enhance the clarity and readability of the English text. No AI tools were employed for data analysis, interpretation, or the generation of original scientific content. The authors take full responsibility for the accuracy and integrity of all scientific aspects of this manuscript.

Conflict of interest

All authors declare that there is no conflict of interest.

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