Effect of either non-encapsulated or encapsulated acidifier-phytobiotic-probiotic on performance, intestinal characteristics and intestinal microflora of local hybrid ducks

Muhammad H Natsir, Osfar Sjofjan, Eko Widodo, Ilham Ardiansah and Eny S Widyastuti¹

Department of Animal Nutrition, Faculty of Animal Science, Brawijaya University, Malang (65145), Indonesia
emhanatsir@ub.ac.id
¹ Department of Animal of Product Technology, Faculty of Animal Science, Brawijaya University, Malang (65145), Indonesia

Abstract

The purpose of this study was to determine the effect of mixture either encapsulated or non-encapsulated of acidifier- phytobiotic-probiotic as a feed additive on production performance, intestinal characteristics and intestinal microflora of local hybrid ducks. The materials used were 192 local hybrid ducks aged 14 d old with average weight 310.28 ± 29.47 grams. Diet was supplemented with a mixture of natural acidifier (lactic acid and citric acid), phytobiotic ( Allium sativum and Phyllanthusiruri), probiotic with encapsulation and non-encapsulation treatment. The research method was a randomized nested 4*2 model design. The treatments were non-encapsulated and encapsulated of acidifier-phytobiotic-probiotic mixtures added at 4 levels (0, 0.5, 1.0 and 1.5 %) with 4 replications (6 ducks in each).

Encapsulation treatment improved body weight gain, feed conversion, the number of Lactic Acid Bacteria (LAB), reduced number of E. coli, improved IOFC, the number and the length of villi. With regard to the level use, this increased body weight gain, improved feed conversion, the number and the length of villi, crypt depth, as well as the number of bacteria (LAB,E coli). It is concluded that the use of feed additives in the form of encapsulated mixture among natural acidifier-phytobiotic-probiotic provide better results than a non-encapsulated form with the optimum level of 1.5 %.

Keywords: acidifier, duck, encapsulated, feed additives phytobiotic, probiotic

Introduction

The local hybrid duck is descended from the Mojosari duck and is well-liked by the Indonesian farmer because it is fast-growing and easily manage. Unfortunately, uncontrolled use of antibiotics feeds as growth promoters (AGPs) have harmful risks for human and ducks. It problem caused using high level of antibiotics to decrease the morbidity, diseases, sickness and mortality. Long term use of antibiotics may cause resistance and leave the residue in the meat and egg product that may be harmful to consumers.

One of the attempts to overcome this problem is going back to nature by utilizing an organic acid as a natural acidifier, which can obtained from the fermentation of molasses (Natsir et al 2010). Extracts from plants such as Phyllanthus niruri and Allium sativum are examples of phytobiotics, which can be used as feed additives to replace synthetic antibiotics. Thus another potential category is the probiotics, such as Lactic acid bacteria (LAB) and yeasts (Zorica et al 2016). A promising approach is to mix acidifiers, phytobiotics and probiotics. It could protected by encapsulation method, so that they pass unchanged through the mouth to the gizzard and intestines which is the area where they will be most effective. The most common method of encapsulating is by spray drying, but it has the disadvantage or requiring high temperatures (140-180 °C) for its process.

The objective of this research reported was to test a simple method of encapsulation using a modified microwave oven at a lower temperature of 60 °C and to determine whether this would improve the effectiveness of a mixed combination of organic acids, phytobiotics, and probiotics in the diet of growing ducks.

Materials and methods

Materials and diets

The research was conducted in Sumbersekar Field Laboratory, Malang, Indonesia. One hundred and ninety-two day-old male hybrid ducks from a local hatchery were used. They had an initial body weight of 43.4 ± 3.32 g/bird. They were randomly allotted to 32 experimental cages of 100 cm x 100 cm x 45 cm size. A mixture of phytobiotics, organic acids, and probiotics was prepared in encapsulated and non-encapsulated form using a modified-microwave oven and a juice extractor.

The sources of phytobiotics (Allium sativum and Phyllanthus niruri L.) were cleaned then blended a with juicer machine. The juices phytobiotics were extracted with ethanol and water (ratio 60:40 w/w). The mixture of phytobiotic was prepared by mixing juice from Allium sativum and Phyllanthus niruri L. in the ratio of 3:1, then BHT (Butylated Hydroxy Toluene) was added at 0.075%. The source of organic acids was prepared by fermenting 60 g pure molasses (within 100 ml buffer pH 7.1) with 2.3 x 10⁹ CFU/ ml Bacillus coagulans in the media, which also contained 5 g yeast extract, 1 ml Tween 89, 2 g (NH₄)₂SO ₄, and 1 g NH₂HPO_4. This fermentation was processed at pH 7.1 and temperature 43^oC for 45 hours.

The combined acid-phytobiotic-probiotic (1:1:1) was mixed with Gum arabic and whey in the ratio of 4:1 at a level of 30%, and then dried using a modified microwave oven equipped with a forced-air blower operating at a temperature of 60 °C.

Experimental design

The research method was a randomized nested 4*2 design. The treatments were non-encapsulated and encapsulated mixtures of an acidifier, phytobiotic, probiotic added at 4 levels: 0, 0.5, 1.0, and 1.5 %, with 4 replications (6 ducks in each). Used feed (Table 1) was free from antibiotics. Diets were fed to appetite with water freely available.

Statistical analysis

The results were analyzed by the ANOVA program in the Genstat Software (Snell and Simpson 1991)

Results

Data on comparation non-encapsulated and encapsulated showed in Table 2. Giving encapsulated mixture among acidifier-phytobiotic-probiotic improved (p< 0.01) on FCR, total colony of LAB and Escherichia coli. But this mixture did not give significant effect on feed consumption and crypt depth. Compared between non-encapsulated and encapsulated mixture among acidifier-phytobiotic-probiotic as feed additive in local hybrid ducks, encapsulated treatment showed better performance on production, characteristics of intestine and intestinal microflora than non-encapsulated tretament.

Figure 1. Effect of level of non- encapsulated or an encapsulated mixture of acidifier- phytobiotic-probiotic on growth performance	Figure 2. Effect of level of non- encapsulated or an encapsulated mixture of acidifier- phytobiotic-probiotic on feed conversion

Figure 3. Effect of level of non- encapsulated or encapsulated mixture of acidifier- phytobiotic-probiotic on Villa height	Figure 4. Effect of level use of either non- encapsulated or encapsulated mixture of acidifier- phytobiotic-probiotic nested on intestinal crypt depth

Figure 5. Effect of level use of either non- encapsulated or encapsulated mixture of acidifier- phytobiotic-probiotic nested on Escherichia coli)

Discussion

A. The effect of level use of non-encapsulated mixture among acidifiers - phytobiotic - probiotic on performances, intestinal characteristics and intestinal microflora of local hybrid ducks

The level use of non-encapsulated mixture among acidifier-phytobiotic-probiotic improved the (p< 0.01) on BWG and villi length (p< 0.01). There was no significant effect on the FC, FCR, IOFC, crypt depth, villi number, and intestinal microflora. Encapsulation could protect citric acid and lactic acid better than non-encapsulated. According to the data, the content of citric acid, essential oils, allicin, phyllanthin and total flavonoid on non-encapsulated mixture of acidifier - phytobiotic were respectively reported as follows: 10.88%, 101.44 mg / 100 g, 11.07 mg/ 100 g and 0.25 mg/ 100g, while the content of citric acid, essential oils, allicin, phylantin and total flavonoids on encapsulated mixture of acidifier-phytobiotic was11.66 %, 694.20 mg/ 100 g, 48.90 mg/ 100 g and 0.27 mg/ 100g respectively. Aksu and Bozkurt (2009) reported that the effect of addition of essential oils, humic acid and essential oil blends - humic acids provide a real influence on feed consumption cumulative broiler until 42 d. Likewise, probiotics do not work to improve feed intake but increase digestibility and health. El-Hakim et al (2009) reported that the use of acidifiers, phytobiotic and a mixture of both of them in broilers did not significantly effect on body weight of broilers 42 d. Furthermore. Isabel and Santos (2009) reported that the addition of acidifier (a mixture of propionic acid and formic acid), essential oils from Syzygium aromathicum and cinnamon extract, as well as acidifier mixed with essential oils extracted from plant, did not show any effect on the body weight of the broiler until the age of 46 days. Thus, Onu (2010) reported broilers which were given Allium sativum, ginger, and a mixture of both could increase the performance of the production. The encapsulation of Origanum virens L. was achieved by the immersion of the aggregates in the essential oil (Ribeiro et al 2004). However, there is a better trend of essential oils and essential oil blends-acidifier use than acidifiers only. The addition of acidifier, phytobiotic extract, and a mixture of both acidifier and extracts phytobiotic have highly significant effect (p< 0.01) toward turkey body weight aged 84 d compared to control. However, among those three types of feed additives have no effect (Mikulski et al 2008).

B. The effect of level use of encapsulated mixture among acidifiers - phytobiotic - probiotic on performances, intestinal characteristics and intestinal microflora of local hybrid ducks

The level use of encapsulated mixture among acidifier-phytobiotic-probiotic showed that encapsulated treatment improved on BWG, FCR, number and length of villi, crypt depth, and LAB significantly (p< 0.01). Importantly, the formulation could reduce the number of Escherichia coli significantly (p< 0.01), while the level use of encapsulated mixture among acidifier-phytobiotic-probiotics does not give significant effect on FC and IOFC. The tendencies effect of level use of either non- encapsulated or encapsulated mixture among acidifier- phytobiotic-probiotic nested on performances, intestinal characteristics and intestinal microflora of local hybrid ducks (Figure 1-5). These results are consistent with Natsir et al (2010) reported that the use of encapsulated lactic acid does not make a significant effect on feed intake than lactic acid liquid, there is even a tendency to decrease feed intake. Ashayerihzadeh et al (2009) reported that the addition of Allium sativum and black cumin did not show a significant effect on the increase of feed intake in broilers. Furthermore, Gauthier (2002) reported that the addition of encapsulated acidifiers could reduce the flow rate of feed that led to slower gastric and then decreased feed consumption. While Gheisari et al (2012) stated that the encapsulated acidifier given to broilers could increase (p< 0.05) Lactobacilli but decrease Coliforms in the ileum of broilers. Natsir et al (2013) reported that encapsulated mixture of Allium sativum and Phyllanthus niruri showed better performance production and characteristics of broiler chicken intestine than in powder form. Gunal et al (2006) stated the use of acidifiers mixture (Genex) in broiler feed decreased (p< 0.05) the number of gram-negative bacteria of 7.95 log CFU/ g (control) to 7.06 log CFU/ g (feed plus acidifier). Yakhkeshi et al (2011) reported that phytobiotic, acidifiers, probiotics, and antibiotics extracts have the ability to reduce pathogenic bacteria in the digestive tract of chicken so that the intestinal health of the chicken could be improved. The results of this research were reinforced by Tollba et al (2010) that reported that the use of mixture between acidifiers and herbal-extract of 2 mg/ kg feed decrease significantly (p< 0.05) the amount of Escherichia coli in chicken intestines compared to the use of herbal extracts at 2 mg/kg.

The mechanism of reduction on pathogenic bacteria in the digestive tract of local hybrid ducks could be explained that acidifier can reduce environmental pH, so that the acidifier can penetrate into the cell walls of pathogens, then resulting in a decrease in pH of bacterial cells. Pathogenic bacteria can not tolerate the decrease of internal and external pH in bacterial cells, so there will be releasing protons H⁺ that requires great energy ATPase pump and this phenomenon lead to bacteria died.Non-pathogenic bacteria can survive and still grow well although the internal pH is low (Gauthier 2002).

Conclusion

• It was concluded that the use of feed additives with encapsulated treatment on mixture among the optimum level of 1.5%. Therefore, using an encapsulated mixture of acidifier-phytobiotic-probiotic at 1.5 % in the feed is recommended.

Acknowledgment

The authors acknowledge financial support from the Ministry of National Education and Culture of Indonesia

Conflict of Interest

We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper.

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Received 30 September 2018; Accepted 25 November 2018; Published 1 January 2019

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