Identification of Chemical Components in Namo-Namo Fruit (Cynometra cauliflora L.) from Halmahera Island and its Potential as a Broiler Chicken Feed Addition

The ban on antibiotic growth promoters (AGP) in the broiler industry has had negative impacts on the growth and health of broiler chickens, threatening the sustainability of broiler production (Prasetyo et al., 2020). To support the sustainability of the broiler industry, the presence of AGP alternatives is urgently needed. Several alternatives to AGP have been recommended, one of which is the use of plant extracts as feed additives. The use of plant extracts as natural additives in poultry feed has increased in recent years, owing to their bioactive components, which are reported to improve production performance, reduce pathogenic bacteria and lower antibiotic residues in meat and eggs (Omar et al., 2016; Jin et al., 2020). According to reports, adding plant extracts to poultry feed can improve meat quality, raise body weight and improve nutrient metabolism by reducing cholesterol and preventing the production of peroxide (Sapsuha et al., 2021; Sugiharto et al., 2022).
       
Halmahera Island is the largest island in North Maluku Province, Indonesia, known for its rich biodiversity. The flora of Halmahera includes various plant species that have long been utilized by the local population, particularly indigenous communities, for traditional medicine (Irmayanti et al., 2022). One of the plant species found on Halmahera Island is Namo-namo (Cynometra cauliflora L.). In Indonesia, the name “namo-namo” is used by the people of Ternate, North Maluku Province, to refer to this plant (Fig 1). However, different regions in Indonesia have their own names for the fruit, such as pukih (Sunda), namu namu (Makassar), puci anggih (Bima) and kuwanjo (Bali). The Namo-namo plant has distinctive features that differentiate it from other plants, including zigzag-shaped branches with an orderly arrangement, leaves that are pale pink when young and glossy dark green when mature and flat coin-shaped seeds located in the center of its flowers, which are pink in color (Suprayogi et al., 2020).


       
Triterpenoids, flavonoids and saponins have been found in ethanol extracts of Namo-namo fruit in earlier research. The ethanol extract of Namo-namo fruit also has antioxidant capacity with an IC50 value of 125.89 ìg/mL and antibacterial activity against foodborne pathogens S. aureus and E. coli, with inhibition zones of 11.43 mm at a concentration of 20% (Ulpiyah et al., 2019; Anliza et al., 2021). The growing location can influence the composition and bioactive content of each plant, leading to variability in its effects on livestock performance and growth (Sugiharto, 2016). To date, the bioactive content and composition of the Namo-namo plant from Halmahera Island, as well as its effects on broiler performance, have not been studied. Based on these issues, it is necessary to conduct research to identify the chemical composition of Namo-namo fruit from Halmahera Island and its potential as a feed additive for broiler chickens.

Preparation of namo-namo fruit extract
 
Namo-namo fruit was collected from the forests of Halmahera Island in North Maluku Province, Indonesia. The fruit was washed with running water, sliced thinly and dried at room temperature. A blender was used to grind the dried fruit into a powder. It is anticipated that the small particle size (powder form) will maximize the extraction of bioactive components by increasing the surface area in direct contact with the solvent. The Namo-namo fruit powder was placed in an Erlenmeyer flask and methanol was added at a 1:4 ratio (w/v). The mixture was macerated at room temperature for three days, with daily stirring. On the fourth day, a rotary vacuum evaporator was used to evaporate the mixture after it had been filtered through Whatman filter paper, yielding a thick Namo-namo fruit extract (Ado et al., 2019).
 
Phytochemical composition and antioxidant activity of namo-namo fruit extract
 
The Namo-namo fruit extract was analyzed for its complete proximate nutrient content based on AOAC (2007). methods. The Folin-Ciocalteu method (Sahreen et al., 2010) was used to assess the total phenol content and spectrophotometry was used to determine the total flavonoids (Mayur et al., 2010). The 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) test was used to measure antioxidant activity (Sochor et al., 2010). Antibacterial activity against E. coli was tested using the disc diffusion (Kirby-Bauer) method.
 
In vivo experiment
 
This study was approved by the Universitas Khairun Faculty of Agriculture’s Animal Ethics Committee (No. 019/KEPH/PH/2024). Between 0 and 7 days of age, 250 unsexed Lohmann broiler chicks were grown together on commercial feed. On day eight, the broilers (147±2.64 g) were divided into five treatments at random, each of which was duplicated five times: T0 (basal feed without Namo-namo fruit extract), T1 (basal feed + 0.25% Namo-namo fruit extract), T2 (basal feed + 0.50% Namo-namo fruit extract), T3 (basal feed + 0.75% Namo-namo fruit extract) and T4 (basal feed + 1% Namo-namo fruit extract). The feed, which came in mash form, was prepared as finisher feed (days 22–35) and starting feed (days 8-21) (Table 1). Up till day 35, food and water were given freely. On day four, all chickens received eye drops of a commercial Newcastle disease (ND) vaccination; on day eighteen, they received drinking water; and on day twelve, they received drinking water containing the Gumboro vaccine. The hens were housed in ventilated broiler houses with sawdust for bedding during the raising phase.


       
Body weight, feed consumption and feed conversion ratio (FCR) were recorded weekly. On day 35, one chick per replicate, with a body weight representing the average of each replicate, was selected, slaughtered, defeathered and eviscerated. The internal organs were taken out and weighed when they were empty. Measurements were also taken of the commercial and broiler carcasses.
               
Analysis of variance was used to statistically evaluate the data (SPSS version 25.0). Duncan’s test was used to identify significant differences between treatments if the treatment had a significant effect (p<0.05) on the measured parameters.

The phytochemical makeup and antioxidant activity of namo-namo fruit extract
 
Natural compounds called phytobiotics or phytogenics are extracted from plants and added to feed to increase the productivity and health of animals. Phytobiotics include various compounds such as essential oils, plant extracts and spices, which possess antimicrobial, antioxidant and anti-inflammatory properties. The use of phytobiotics in the livestock industry is increasingly popular due to their ability to replace antibiotics as growth promoters (Sugiharto 2016). The phytochemical makeup and antioxidant capacity of the Namo-namo fruit extract from Halmahera Island are displayed in Table 2. Phytochemical analysis indicated phenol and flavonoid contents of 1.98 g/100 g and 2.01 g/100 g, respectively. Our study also showed that Namo-namo fruit from Halmahera Island exhibited antioxidant activity with an IC50 of 4.72 (2,2-diphenyl-1-picrylhydrazyl hydrate, DPPH).


       
The Namo-namo fruit from Halmahera Island has an IC50 for antioxidant activity and contains bioactive substances like flavonoids and phenols. These results are in line with earlier research showing that the Namo-namo plant exhibits potent antioxidant activity and phytochemical content (Abd Aziz et al., 2017; Suprayogi et al., 2020). The bioactive compounds in plants are reported to be highly beneficial for animal health and productivity. By enhancing digestive enzymes and the absorption of vital nutrients, certain bioactive substances-like saponins, tannins and flavonoids-can improve digestive efficiency and have a good effect on cattle growth (Bera et al., 2019; Perin et al., 2019; Zhang et al., 2017).
       
The results of the antibacterial inhibition test of the Namo-namo fruit extract using the disc diffusion method (Kirby-Bauer) against E. coli growth showed an inhibition zone of 12.16 mm (Table 2), indicating that the Namo-namo fruit extract has moderate inhibitory activity against E. coli growth. The study by Ulpiyah et al., (2019) demonstrated that the leaves of the Namo-namo fruit could inhibit the growth of the pathogenic bacterium Porphyromonas gingivalis, while the study by Abd Wahab et al., (2019) showed that ethanol extracts of C. cauliflora leaves could serve as a potential antibacterial agent due to the presence of active antibacterial compounds.
 
Performance of broiler chicken growth
 
According to this study, broiler chicken body weight gain was significantly (p<0.05) increased when Namo-namo fruit extract from Halmahera Island was added to diet (Table 3). There are currently no studies in the literature that describe how Namo-namo fruit extract affects the body weight gain of broilers. Nonetheless, the synergistic activity of the fruit’s many phytochemicals, including flavonoids and phenols (Table 2), is probably what makes Namo-namo fruit extract so powerful. This, in turn, enhances the physiological circumstances of the hens. According to reports, phytoche-micals including phenols and flavonoids support the preservation of the microbial equilibrium in the broiler’s digestive system, thereby enhancing nutrient absorption and chicken health, which leads to better feed utilization and efficiency. Moreover, previous studies have shown that Namo-namo fruit has antibacterial properties (Ulpiyah et al., 2019), antifungal properties (Suprayogi, 2020), antiviral properties (Abd Wahab et al., 2021), antimicrobial properties (Abd Wahab et al., 2019) and antioxidant properties (Abd Aziz et al., 2017; Ado et al., 2019; Samling et al., 2022), which can improve the development performance of broiler chickens by promoting the growth of good bacteria, deactivating harmful bacteria and speeding up metabolism and nutrient absorption in the digestive tract.


       
Better feed usage is demonstrated by the increase in feed conversion seen in broiler chickens fed Namo-namo fruit extract from Halmahera Island as opposed to the control group (T0). better digestion, especially protein digestion, through higher digestive enzyme activity is probably the cause of the better feed conversion in broilers fed Namo-namo fruit extract (Salgado-Tránsito et al., 2011). Because the bioactive chemicals in plant extracts lead to increased body weight in chickens, previous research has suggested using plant extracts as phytobiotics in broiler feed to boost growth (Basit et al., 2020; Zaikina et al., 2022). The use of Namo-namo fruit extract may enhance feed consumption, which in turn can lead to increased body weight. The increase in feed consumption is likely due to the flavonoid and phenol content in the Namo-namo fruit extract, which can improve the flavor of the feed, thereby increasing its palatability and stimulating the chickens’ appetite (Sugiharto, 2016; Sapsuha et al., 2022). An interesting finding was observed regarding the feed conversion ratio (FCR) of broilers. In this study, the inclusion of 1% Namo-namo fruit extract in the feed significantly (p<0.05) reduced the FCR of broilers, indicating improved feed efficiency. This suggests that feeding broilers with Namo-namo fruit extract from Halmahera Island does not adversely affect the digestibility of the chickens.
 
Weight of internal organs and characteristics of the broiler chicken carcass
 
The weight of the internal organs of broiler chickens was not significantly affected (p>0.05) by the administration of Namo-namo fruit extract from Halmahera Island, according to the results (Table 4). This indicates that the use of Namo-namo fruit extract does not affect the health of broilers. The relative weight of the internal organs of broiler chickens was unaffected by the addition of telang flower extract to feed, according to Sapsuha et al., (2023), who discussed the use of herbal plants in feed. Age, strain type and environmental factors like feed and management techniques can all affect how much weight the internal organs of broilers weigh. According to other research, giving broilers cashew, nutmeg pulp, or bilimbi had no influence on the relative weight of their internal organs (Mareta et al., 2020; Sapsuha et al., 2021; Martínez et al., 2021).


               
According to the study’s findings, broiler chickens’ carcass % and commercial carcass cuts were unaffected (p>0.05) by the addition of Namo-namo fruit extract from Halmahera Island (Table 5). According to Mareta et al., (2020), there were no appreciable changes in the carcass weight or commercial carcass cuts of broiler chickens when bilimbi was added to the diet. According to Vase-Khavari et al., (2019), a different study that used three medicinal plants-R. coriaria, H. persicum and M. piperita-also found no discernible impact on the commercial carcass cuts and carcass weight of broilers.

Because it enhances performance without negatively impacting the carcass features of broiler chickens, the Namo-namo fruit extract from Halmahera Island, which includes bioactive substances such phenols and flavonoids, can be utilized as a feed addition.
 

The Republic of Indonesia’s Ministry of Education and Culture’s Directorate of Research and Community Service, Directorate General of Higher Education, provided financial assistance for the study under Contract No. 055/E5/PG.02.00.PL/2024.
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.
 
Informed consent
 
The Committee of Experimental Animal Care and the Animal Ethics Committee of the Faculty of Agriculture, Universitas Khairun, approved all animal procedures for experiments (No. 019/KEPH/PH/2024).
 

The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.