Dasar Nutrisi Ruminansia

PAZLA et al. Effects of supplementation during oil palm frond fermentation 1837

found for the T3 treatment (P> 0.05) compared with the T1 REFERENCES
treatment, but the content from the T3 treatment did not
differ significantly (P <0.05) from that of the T2 treatment. AOAC. 2010. The Official Method of Analysis of the Association of
The low content of CF in the T3 treatment with 2000 ppm
P is assumed to be capable of triggering growth and Official Analytical Chemists. Washington D.C., USA.
extension of the fungus mycelia, thereby resulting in a
reshuffle in the fermentation palm cell wall section. Cell BPS. 2015. Statistik Kelapa Sawit Indonesia (Indonesian Oil Palm
wall reshuffling occurs during the fermentation process due
to the role of Phanerochaete chrysosporium, which utilizes Statistics) 2015. BPS Indonesia, Jakarta. [Indonesian]
cell contents as nutrients for growth. According to Tuomela
et al. (2000), the utilization of soluble material occurs in Brook EJ, Stanton WR, Wall-Bridge A.1969. Fermentation methods for
the early phase of fermentation, and the reshuffle of cell
wall components occurs as required by the mold (Suparjo protein enrichment of cassava. Biotechnol Bioeng 11: 1271-1284.
and Nelson 2012). The greater the dose of P used, the
greater the decrease observed in the CF content. In the Elihasridas. 2012. The effect of supplementation of mineral Zinc on in
fermentation process, microorganisms need macro- and
micronutrients for growth. One of the required vitro digestibility of amoniated corn cobs ration. J Peternakan 9: 9-14.
macronutrients is phosphorus (Pazla et al. 2018). The
process of glycolysis produces chemical energy in the form [Indonesian]
of ATP compounds that contain high-energy phosphate
groups because glycolysis is instrumental in the process of Fadilah, Distantina S, Dwiningsih SR, DS. 2009. Pengaruh
phosphorylation through the transfer of radical phosphate
groups during metabolism (Rodwell et al. 2019). In penambahan glukosa dan ekstrak yeast terhadap biodelignifikasi
addition, in the cell, phosphorus is present as a
phosphoprotein, phospholipid and nucleic acid compound. ampas batang aren. Ekuilibrium 8 (1): 29-33. [Indonesian]
The combination of Ca, P and Mn minerals in the T3
treatment succeeded in producing optimum mold growth, Febrina D. 2016. Pemanfataan Biodelignifikasi Pelepah Sawit
enabling enzyme production to be even greater so that the
mold work process in degrading the cell wall will increase. menggunakan Kapang Phanerochaete chrysosporium sebagai Pakan
Febrina (2016) stated that the optimal combination of Ca
and Mn minerals can produce optimal ligninase enzymes as Utama Ternak Ruminansia. [Disertasi]. Program Pasca Sarjana,
well as break down the cell wall in the fermentation
process. Fakultas Peternakan, Universitas Andalas, Padang. [Indonesian]

The P (2000 ppm), Ca (2000 ppm) and Mn (150 ppm) Gassara F, Brar SK, Tyagi RS, Verma M, Surampalli RY. 2010.
minerals in T3 give the best results in increasing the supply
of the ligninase enzyme to achieve a decrease in lignin Screening of agro-industrial wastes to produce ligninolytic enzymes
content during OPF fermentation. This result has positive
implications for the digestibility of cellulose and by Phanerochaete chrysosporium. Biochem Eng J 49: 388-394.
hemicellulose by rumen microbes on the palm frond. The
main challenge to the implementation of this research is the Imsya A. 2013. Hasil Biodegradasi Lignoselulosa Pelepah Kelapa Sawit
fermentation process, which is difficult to carry out in rural
areas. (Elaeis quineensis) oleh Phanerochaete chrysosporium sebagai

To conclude, supplementation of fermented OPFs by Antioksidan dan Bahan Pakan Ternak Ruminansia. [Disertasi].
Phanerochaete chrysosporium with treatments of Ca (2000
ppm), P (2000 ppm) and Mn (150 ppm) in T3 provided the Sekolah Pasca Sarjana, Institut Pertanian Bogor, Bogor. [Indonesian]
lowest crude fiber content, the highest ligninase enzyme
activity, a decreased lignin content, and the highest crude Jamarun N, Arief, Satria B. 2016. Pemanfaatan Limbah Kebun dan
protein content.
Limbah Industri Kelapa Sawit Supplementasi Probiotik pada Ransum
ACKNOWLEDGEMENTS
Kambing Peranakan Etawa Menunjang Program Swasembada Susu
This research was funded by the Andalas University
Professor Grant 2016-2018. We thank the staff of the 2020. Laporan MP3EI. Kementerian Pertanian, Jakarta. [Indonesian]
technology laboratory and the feed industry of the Faculty
of Animal Science, Andalas University, Padang, Indonesia Jamarun N, Zain M, Arief, Pazla R. 2017. Effects of calcium, phosphorus
and the staff of the dairy nutrition laboratory of the IPB
University, Bogor, Indonesia. and manganese supplementation during oil palm frond fermentation

by Phanerochaete chrysosporium on laccase activity and in vitro

digestibility. Pak J Nutr 16: 119-124. DOI: 10.3923/pjn.2017.119.124

Jamarun N, Zain M. 2013. Dasar Nutrisi Ruminansia. Penerbit Jasa Surya,

Padang. [Indonesian]

Jamarun N, Zein M, Arief, Pazla R. 2018. Populations of rumen microbes

and the in vitro digestibility of fermented oil palm fronds in

combination with Tithonia ( Tithonia diversifolia ) and elephant grass

(Pennisetum purpureum). Pak J Nutr 17: 39-45. DOI:

10.3923/pjn.2018.39.45

JamarunN, Agustin F, Pazla R, Cahyani O. 2017. N. Effects of phosphor

(P) supplementation in combination with calcim (ca) and manganese

(mn) during oil palm frond fermentatation by Phanerochaete

chrysosporium on fiber fraction content. Proceedings of The 5th

International Seminar of Animal Nutrition and Feed Science 7-9

Nopember. Mataram, Indonesia.

Jonathan SG, Fasidi IO, Ajayi AO, Adegeye A. 2008. Biodegradation of

Nigerian wood waste by Pleurotustuber-regium (Fries) Singer.

Bioresour Technol 99: 807-811.

Martínez AT, Speranza M, Ruiz- Dueñas FJ, Ferreira P, Camarero S,

Guillén F. 2005. Biodegradation of lignocellulosics: microbial,

chemical, and enzymatic aspects ofthe fungal attack of lignin. Int

Microbiol 8: 195 204.

Musnandar E. 2003. Pengaruh dosisinokulum Marasmius sp. dan lama

inkubasi terhadap kandungan komponen serat dan protein murni pada

sabut kelapa sawit untuk bahan pakan ternak. Jurnal Ilmiah Ilmu-Ilmu

Peternakan 9(4): 225-234.

Nelson, Suparjo. 2011. Penentuan lama fermentasi kulit buah kakao

dengan Phanerochaete chrysosporium: Evaluasi kualitas nutrisi

secara kimiawi. Agrinak 1(1): 1-10. [Indonesian]

Pasaribu T, Arini N, Purwadaria T, Sinurat AP. 2003. Evaluasi nilai gizi

fermentasi lumpur sawit dengan penambahan fosfor dari sumber yang

berbeda. JITV 8(3): 157-163. [Indonesian]

Pazla R, Jamarun N, Zain M, Arief. 2018. Microbial protein synthesis and

in vitro fermentability of fermented oil palm fronds by Phanerochaete

chrysosporium in combination with tithonia (Tithonia diversifolia)

and elephant grass (Pennisetum purpureum). Pak J Nutr 17: 462-470.

DOI: 10.3923/pjn.2018.462.470.

Pazla R, Zain M, Ryanti I, Dona A. 2018. Supplementation of minerals

(phosphorus and sulfur) and Saccharomyces cerevisiae in a sheep diet

based on a cocoa by-product. Pak J Nutr 17: 329-335. DOI:

10.3923/pjn.2018.329.335.

Perez J, Dorado JM, Rubia T, Martinez J. 2002. Biodegradation and

biological treatment of cellulose, hemicellulose, and lignin; an

overview. Int Microbiol 5: 53-63.

287Dasar Nutrisi Ruminansia (Edisi ke II)

1838 21 (5): 1833-1838, May 2020

Rodwell VW, Bender D, Botham KM, Kennelly PJ, Weil PA. 2019. enriched ion Mn2+ and Ca2+. [Dissertation]. Bogor Agricultural
Harper's Illustrated Biochemistry. 31st ed. McGraw-Hill Education / Institute, Bogor. [Indonesian]
Medical, New York. Suryani H, Zain M, Ningrat RWS, Jamarun N. 2016. Supplementation of
Direct Fed Microbial (DFM) on in vitro fermentability and
Singh D, Chen S. 2008. The White-rot fungus Phanerochaete degradability of ammoniated palm frond. Pak J Nutr 15(1): 89-94.
chrysosporium conditions for the productions if lignin degrading DOI:10.3923/pjn.2016.89.94
enzymes. Appl Microbiol Biotechnol 81: 399-417. Tuomela M, Vikman M, Hattaka A, Itavaara M. 2000. Biodegradation of
lignin in a compost environment: A review. Biosresour Technol 72:
Subagiyo, Margino S, Triyanto. 2015. Pengaruh penambahan berbagai 169-183.
jenis sumber karbon, nitrogen dan fosfor pada medium deMan, Van Soest PJ, Robertson JB, Lewis BA. 1991. Methods for dietary fiber,
Rogosa and Sharpe (MRS) terhadap pertumbuhan bakteri asam laktat neutral detergent fiber and nonstarch polysaccharides in relation to
terpilih yang diisolasi dari istestinum udang penaeid. Jurnal Kelautan
Tropis 18 (3): 127-132. [Indonesian] animal nutrition. J Dairy Sci 74: 3583-3597.
Zain M, Jamarun N, Tjakradidjaja AS. 2010. Phosphorus supplementation
Suparjo, Nelson. 2012. Fraksi serat dan kecernaan in vitro kulit buah
kakao yangdifermentasi dengan Phanerochaete chrysosporium. of ammoniated rice straw on rumen fermentability, synthesised
Agrinak 2 (1): 41-48. [Indonesian] microbial protein and degradability in vitro. World Acad Sci Eng
Technol 4: 357-359.
Suparjo. 2008. Degradasi komponen lignoselulosa oleh kapang pelapuk
putih. jajo66.wordpress.com. [Indonesian]

Suparjo. 2010. Fruit Leather Nutritional Quality Improvement Cocoa Pod
Husk as Feed in Bioprocess by Phanerochaete chrysosporium

288 Dasar Nutrisi Ruminansia (Edisi ke II)

289Dasar Nutrisi Ruminansia (Edisi ke II)

290 Dasar Nutrisi Ruminansia (Edisi ke II)

291Dasar Nutrisi Ruminansia (Edisi ke II)

292 Dasar Nutrisi Ruminansia (Edisi ke II)

293Dasar Nutrisi Ruminansia (Edisi ke II)

294 Dasar Nutrisi Ruminansia (Edisi ke II)

Quest Journals
Journal of Research in Agriculture and Animal Science
Volume 7 ~ Issue 1 (2020) pp: 31-34
ISSN(Online) : 2321-9459
www.questjournals.org

Research Paper

Effect of Biourine and Fungi Mycorrhiza arbuscular on
Production and In-vitroNutrient Digestibility of Kumpai grass
(Hymenachneamplexicaulis(Rudge) Nees) planted at Ex-coal Mine

Land as Animal feed.

Hardi Syafria1), Novirman Jamarun2)and Roni Pazla2

1Faculty of Animal Science, Jambi University, Jambi,
2 Faculty of Animal Sciece AndalasUniversity, Limau Manis, Padang.

ABSTRACT
The aim of this study is to show the effect of biourine and fungi Mycorrhiza arbuskula (FMA) on production and
in-vitro nutrient digestibilityof Kumpai grass (Hymenachneamplexicaulis (Rudge) Nees) planted at ex-coal mine
land as animal feeds. The experimental design was completely randomized with 4 treatments and 5 replications
per treatment. The treatment consists of: A, concentration of 0% Biourine + FMA 20 g/pot, B, 15% Biourine +
FMA 20 g/pot, C, 30% Biourine+ FMA 20 g/pot and D, 45% Biourine + FMA 20 g/pot. Variables observedare
grass production, dry matter and crude protein content, percentage of root infections by Mycorrhiza arbuscula,
and in-vitro dry and organic matter digestibility. The results of the experiment indicated that the treatments had
a very significant effect (P<0.01) on all observed parameters. Treatment D, 45% biourin + FMA 20 g/pot gave
the best result, followed by the treatment C, 30% biourine + FMA 20 g/pot, and treatment B, 15% biourine +
FMA 20 g/pot and the lowest value was in treatment A, 0% biourine + FMA 20 g/pot. From the results, it can be
concluded that the treatment D, 45% biourine + FMA 20 g/pot was the best treatment;it produced the highest
value for all observed variables.
KEYWORD: Biourin, Mycorrhiza, Kumpai grass, digestibility, ex-coal land.

Received 09 October, 2020; Accepted 24 October, 2020 © The author(s) 2020.
Published with open access at www.questjournals.org

I. INTRODUCTION
Kumpai grass (Hymenachneamplexicaulis (Rudge) Ness), is a natural resource of high biological value
in Indonesia especially in Jambi province. It has the potential to support the availability of adequate forage for
ruminant feeds based on local resources [1]. Available land for forage planting usually decreased with time,
because fertile land is mainly used for food crops, plantations and even non-agricultural needs [2. The ex-coal
mine area in Jambi province is advancing further, due to the increased mining activities. Hundreds or even
thousands of hectares of land have been rendered unproductive due to structural damage and degradation of soil
nutrients. Therefore, use of land productively is very necessary. The utilization of biourin and Mycorrhizae
arbuscula as an agent of biotechnology for increasing productivity of ex-coal mine land is a viable alternative that
requires research. Therefore, this study aims to find the effect of utilization of biourin and Mycorrhizae arbuscula
fungi on the production, in-vitro nutrient digestibility, and nutrient composition of kumpai grass
(Hymenachneamplexicaulis(Rudge) Nees) planted in ex-coal mine land as animal feed.

II. MATERIALS AND METHODS
The study was conducted in Kotabaru city, District of Jambi for 5 (five) months. Forages were analyzed
at the laboratory of Animal Nutrition, Faculty of Animal Husbandry University of Jambi, and in-vitro nutrient
digestibility analyzed at Ruminant Nutrition Laboratory, Faculty of Animal Science, Andalas University.

Material and Equipment
The ex-coal mine land was used as a planting medium, 5 kg / pot each, kumpai grass, fungi ofMycorrhiza

arbuscular of multiple spore types (Glomus sp, Acaulosporasp and Scutellosporasp,.), and biourine. The
equipment used include tillage tools, lawn mowers, rule, sprinklers, plastic bags, scales, and apparatus for

Corresponding Author: Hardi Syafria 31 | Page

295Dasar Nutrisi Ruminansia (Edisi ke II)

Effect of Biourine and Fungi Mycorrhiza arbuscular on Production and In-vitroNutrient ..

analyzing forage.

Research methods
The experiment applied a completely randomized design (CRD), with four treatments and five

replications per treatment. Each treatment consists of: A, Concentration of 0% biourine+ FMA 20 g/pot, B,
Concentration of 15% biourine+ FMA 20 g/pot, C, Concentration of 30% biourine+ FMA 20 g/pot, and D,
Concentration of 45% biourine+ FMA 20 g/pot.

Observed variables
Variables observed include dry matter production and content, crude protein,in-vitro digestibility of both

dry and organic matter and percentage of roots infections from Mycorrhiza arbuscula fungi.

Research Implementation
Before the kumpai grass (Hymenachneamplexicaulis(Rudge) Nees) were planted, the planting soil was

placed into a composite medium of depth of 0-20 cm. Next, it was air-dried and cleaned, with unneeded materials
such as plant roots removed. The use of Mycorrhiza arbuscular fungi as a treatment was based on results of [3]
research, which used 20 g/pot. Biourin, used in the experiment was aerated for 6 hours and then fermented for 21

days. Two weeks before planting, a polybag was prepared and filled with soil at 5 kg/pot.Mycorrhizae arbuscula
was then released into the grass within the pot, by inserting an inoculum into each planting hole, while Biourine
was administered when the grasses reached two weeks old.

Data processing
Data was processed in a completely randomized design and analysis of variance (ANOVA) was used

were used to observe for any significant effects.

III. RESULTS AND DISCUSSION
Dry Matter Production
The results of the experiment on dry matter production at first and second cutting of Kumpai grass
(Hymenachneamplexicaulis(Rudge) Nees) plantedat ex-coal mine landare shown in Table 1.

Table1.Effect of application of Biourine and Fungi of Mycorrhizae arbuscular on dry matter production at

first and second cutting of Kumpai grass (g/pot).

Treatments Dry matter production

First cutting Second cutting

A 50.60 d 56.60 d

B 65.10 c 70.25 c

C 72.40 b 75.60 b

D 78.10 a 82.34 a

The numbers in the same row followed by different letters are significantly different (P<0.05).

The Production of Kumpai grass at the first and second cutting periods with application of 45% biourine
+ FMA 20 g/pot (treatment D) showed the highest production (P <0.05), compared to the other treatments. The
average dry matter production during the second cutting period went higher than the first (for all treatments).
Treatment D (45% Biourine + FMA 20 g / pot) showed a better plant growth compared with the others, in both
first and second cutting periods. This is because Mycorrhizae arbuscularequires an organic fertilizer for nutrition
and energy. Also, oxygen consumption was increased, enabling the plants to absorb more mineral salts and
hydrogen ions which can be exchanged. [4] stated that Fungi Mycorrhiza arbuscula increases the absorption of
nutrients and water from the soil, thus enabling these plants to reproduce new cells and hormones to improve plant
growth and soil aggregates. It also speeds up the process of mass flow. In the same climatic conditions, soil fertility
exerts more influence on growth and development of plant cells [5]. [6] indicated that application of FMA and
organic fertilizer (compost, cow dung, etc) in ultisol soil can increase production of grass dry matter.

In-vitro Dry Matter and Organic Matter digestibility.
The average in-vitro digestibility of dry matter and of organic matter ofKumpai grass (Hymenachneamplexicaulis
(Rudge) Ness) in this study are shown in Table 2.

Corresponding Author: Hardi Syafria 32 | Page

296 Dasar Nutrisi Ruminansia (Edisi ke II)

Effect of Biourine and Fungi Mycorrhiza arbuscular on Production and In-vitroNutrient ..

Table2.Effect of application Biourine and FMA on in-vitro dry matter and organic matter digestibility of

Kumpai grass (Hymenachneamplexicaulis(Rudge) Nees).

Treatment Dry matter In-vitro digestibility (%)
Organic matter

A 50.43 c 48.43 c

B 51.54 c 49.15 c

C 54.58 b 51.14 b

D 57.13 a 55.24 a

The numbers in the same row followed by different letters are significantly different (P<0.05)

Increasing in-vitro dry and organic matter digestibility of Kumpai grass (Hymenachneamplexicaulis
(Rudge) Ness) occur due to the ability of Mycorrhizae arbuscula to increase the absorption of nutrients and water
from the soil. This results in increased plant growth and in turn, affects the digestibility of dry and organic matter.
Dry matter contains organic matter, therefore if dry matter digestibility increases, the same occurs for that of
organic matter. [7] states that in-vitro digestibility of dry matter is directly proportional to that of organic matter;
Increased digestibility of dry matter indicates increased digestibility of organic matter. Anotherfactor that helps
to boost digestibility is the growth and development of soil microorganisms; this usually begins the process of
nitrogen transformation. In the soil, nitrogen transformation effects the conversion of organic nitrogen into
inorganic forms that are absorbable by plants [8].

Crude protein content.
The results of the analysis of variance showed that the treatment had a significant effect (P<0.01) on

crude protein content of Kumpai grass (Hymenachneamplexicaulis (Rudge) Ness). The average forage protein
content in the study is listed in Table 3.

Table 3. Effect of application of Biourine and Fungi of Mycorrhizae arbuscular on Crude

Protein content of Kumpai grass (Hymenachneamplexicaulis(Rudge) Nees).

Treatment Crude Protein (%)

A 12,10 c

B 13,65 b

C 15,20 a

D 15,35 a

The numbers in the same row followed by different letters are significantly different (P<0.01)

Increasing the addition of Biouring increases the crude protein content of Kumpai grass
(Hymenachneamplexicaulis(Rudge) Nees).This is because the hypha from Mycorrhiza arbuscula associated with
plant roots help in the absoprtion of water and nutrient from soil pores. Mycorrhiza arbuskula infects the root
system by forming hyphae intensively to increase nutrient uptake, especially phosphorus mineral for carbohydrate
metabolism. Also, it improves soil structure which enhances the development of plant roots, thus affecting the
quality or crude protein of the Kumpai grass. Spores of Mycorrhizae arbuskula contain nitrate reductase, therefore
its external hyphae contain a nitrate absorption capacity [9](Bago et al., 1996). External hyphae of the fungi can
also increase the absorption of N, Ca and Mg from the soil [10], including that of micro minerals such as Zn, Cu,
and B [11].

Kumpai Roots Infection by Mycorrhizae arbuscula.
Results of the analysis of variance showed that the treatments had an effect (P <0.01) on the percentage of root

infection of Kumpai grass by Mycorrhizae arbuscula as shown. in Table 4.

Table 4. Effect of application of Biourine and Fungi Mycorrhizae arbusculae on roots infection of

Kumpai grass (Hymenachneamplexicaulis(Rudge) Nees).

Treatment Roots Infection (%)

A 20 c

B 32 b

C 47 a

D 50 a

The numbers in the same row followed by different letters are significantly different (P<0.05)

Corresponding Author: Hardi Syafria 33 | Page

297Dasar Nutrisi Ruminansia (Edisi ke II)

Effect of Biourine and Fungi Mycorrhiza arbuscular on Production and In-vitroNutrient ..

The increasing root infections of Kumpai grass (Hymenachneamplexicaulis (Rudge) Ness) at treatment
D occurred due to increased administration of Biourin which enhanced the availability of carbohydrate and
phosphorus in supporting the optimal root infection process. It was discovered that the roots structure contained
vesicles, spores and hyphae which characterizes root infection by mycorrhiza. Fungi of Mycorrhiza arbusculae
infect plant root systems by forming external hyphal braids, thereby increasing root capacity and rate of absorption
from the soil [12]. Mycorrhizae roots, when bound with hyphae (knownas root hairs; rhizomorf) absorb available
nutrients and water from the soil. However, hyphae are widely spread in the soil, which boosts and enhances
absorption.

IV. CONCLUSION
The treatment of D (concentration of 45% biourine and FMA 20 g/pot) was successful, as it showed thehighest
dry matter production, and in-vitro dry and organic matter digestibility of kumpai grass

(Hymenachneamplexicaulis(Rudge) Nees) compared to the others treatments.

ACKNOWLEDGMENTS

The author appreciates the Directorate of Research and Community Service, Ministry of Research, Technology

and Higher Educationfor the second quarter of 2019.

REFERENCES

[1]. Syafria. H. 2016. Increase of result and nutritional value of kumpai grass (Hymenachneamplexicaulis (Rudge) Nees.) with fungi of
mycorrhiza arbuscular and organic fertilizer in Ultisol as animal feed. Dissertation. Pos-tgraduate of Andalas University. Padang.

[2]. Jamarun, N. dan Mardiati Zain. 2013. Basic Ruminant Nutrition. Publisher ofJasa Surya Padang.
[3]. Syafria. H. 2016. Increase of result and nutritional value of kumpai grass (Hymenachneamplexicaulis (Rudge) Nees.) with fungi of

mycorrhiza arbuscular and organic fertilizer in Ultisol as animal feed. Dissertation. Pos-tgraduate of Andalas University. Padang
[4]. Beinroth, F. H. 2001. Land resources for forage production in the tropics. In Sotomayor-Rios A. Pitman Wd (eds) Tropical Forage

Plants Development and Use .CRC Press. P 3 - 15.

[5]. Syafria. H. 2009. Effects of nitrogen fertilization and spacing on growth and production of local kumpai grass
(Hymenachneamplexicaulis (Rudge) Nees.). Scientific Magazine of Percikan Bandung. Edition on May 2009. ISSN: 0854 - 8986.

page: 97-100. Bandung
[6]. Syafria. H. 2016. Increase of result and nutritional value of kumpai grass (Hymenachneamplexicaulis (Rudge) Nees.) with fungi of

mycorrhiza arbuscular and organic fertilizer in Ultisol as animal feed. Dissertation. Pos-tgraduate of Andalas University. Padang
[7]. Sutardi, T. 1980. The cornerstone of nutrition science. DepartemenIlmu Department of Animal Food Sciences Faculty of Animal

Husbandry, Bogor Agricultural University, Bogor
[8]. Widjajanto, D.W., Honmura, T., Matsushita, K., and Miyauchi, N. 2001. Studies on the release of N from waterhyacinincorporated

intosoil-crop system susing15N- labeling techniques. Pak.J. Biol. Sci.,4 (9): 1075 - 1077.
[9]. Bago, B., H. Vierheilig, Y. Piche, C. Azcon-Aguilar. 1996. Nitrat Depletion and pH Changes Induced by the Extraradica Mycelium

of the Arbuscular Mycorrhizal Fungus Glomus Intraradices in Monoxenic Culture. New Phytol.133:273-280
[10]. Hapsoh. 2008. Utilization of FMA in Soy Cultivation in Dry Land.University of

Sumatera Utara.
[11]. Smith, S. E. and D. J. Read. 2008. Mycorrhizal Symbiosis. Third edition: Academic Press. Elsevier Ltd. New York, London,

Burlington, San Diego. 768p.

[12]. Cruz, C., J. J. Green, C.A. Watson, F. Wilson, and M.A. Martin-Lucao. 2004. Functional aspect of root architecture and mycorrhizal
inoculation with respect to nutrient uptake capacity. Mycorrhiza 14:177-184

Hardi Syafria, et. al. "Effect of Biourine and Fungi Mycorrhiza arbuscular on Production and In-
vitroNutrient Digestibility of Kumpai grass (Hymenachneamplexicaulis(Rudge) Nees) planted at
Ex-coal Mine Land as Animal feed." Quest Journal of Research in Agriculture and Animal Science,
vol. 07, no. 01, 2020, pp. 31-34.

Corresponding Author: Hardi Syafria 34 | Page

298 Dasar Nutrisi Ruminansia (Edisi ke II)

Volume 21, Number 11, November 2020 ISSN: 1412-033X
Pages: 5230-5236 E-ISSN: 2085-4722
DOI: 10.13057/biodiv/d211126

Chemical composition and rumen fermentation profile of mangrove
leaves (Avicennia marina) from West Sumatra, Indonesia

NOVIRMAN JAMARUN1,2, , RONI PAZLA1,3, , ARIEF1, ANURAGA JAYANEGARA4, GUSRI YANTI1

1Department of Nutrition Science and Feed Technology, Faculty of Animal Husbandry, Universitas Andalas. Jl. Raya Unand, Limau Manis, Padang
25163, West Sumatra, Indonesia. Tel./fax.: +62-751-71464, email: [email protected], [email protected]

2Department of Education Quality Assurance, Faculty of Medical, Universitas Baiturrahmah. Jalan Raya By Pass Km. 14, Aie Pacah, Padang 25586,
West Sumatra, Indonesia

3Department of Extension Science and Development Communication, Graduate Program, Universitas Andalas. Jl. Raya Unand, Limau Manis, Padang
25163, West Sumatra, Indonesia.

4Department of Nutrition Science and Feed Technology, Faculty of Animal Husbandry, Institut Pertanian Bogor . Jl. Agatis, Kampus IPB Darmaga,
Bogor 16680, West Java, Indonesia

Manuscript received: 16 September 2020. Revision accepted: 16 October 2020.

Abstract. Jamarun N, Pazla R, Arief, Jayanegara A, Yanti G. 2020. Chemical composition and rumen fermentation profile of mangrove
leaves (Avicennia marina) from West Sumatra, Indonesia. Biodiversitas 21: 5230-5236. This study aimed to determine the potential of
mangrove leaves of Avicennia marina for ruminant animal feed. Laboratory tests were carried out on A. marina with three replicates.
Parameters measured were proximate and fiber contents, rumen fluid profile (pH, NH3 and VFA), digestibility of nutrients (DM, Ash,
CP, CF, NDF, ADF, cellulose, and hemicellulose), macro and micro mineral contents, and phytochemical compounds. The results
showed the nutritional content of A. marina were CP 13.37%; Ash 7.17%; lignin 7.34%; TDN 79%, rumen fluid profile is in reasonable
condition, digestibility of food substances is more than 50%, rich in macro and micro minerals and contains phytochemical compounds
such as phenols, steroids, triterpenoids, and tannins. Macro and micro minerals content of Ca 0.38%, Na 0.20%, Mg 0.20%, K 0.48%, P
0.51%, S 0.01%, Cl 1.03%, Fe 388 ppm, Zn 164 ppm, Mn 211 ppm, and Cu 128 ppm. This research concludes that A. marina is very
potential to be used as a ruminant animal feed.

Keywords: Avicennia marina, mangroves, minerals, phytochemicals, proximate, rumen fluid

INTRODUCTION 1.5-2.0 cm (Kitamura et al. 1997). In the coastal areas of
Indonesia, people use their leaves to feed goats. These
Indonesia is a country with the most extensive leaves fall off, and the amount is quite adequate as a forage
mangrove forests globally (Richards and Friess 2016; source for animal feed. Nevertheless, to date, there is little
Bunting et al. 2018). Indonesia's reliable mangrove forests research that explores the potential of A. marina leaves as
are currently 3,361,216.61 ha (Rahardian et al. 2019). ruminant feed. This study aimed to evaluate A. marina
Mangrove forests help to reduce the impact of hurricanes, leaves' possibility as ruminant feed in terms of nutritional
large waves, and winds from tropical cyclones. Mangrove content, phytochemicals, digestibility, and rumen fluid
trees reduce wave energy as they pass through mangrove profile in vitro. This research held on Sebelas Tarusan Sub-
forests and become barriers between streams and land district, Pesisir Selatan District, Province of West Sumatra,
(United Nations Environment Program 2014). When the Indonesia. The mangrove area mostly consists of A.
sea is high tide, mangrove forests are flooded with water, marina, but there are also Avicennia alba plant clusters in
and at low tide, thick mud covers the surface of the soil, some locations.
which stores wealthy organic material (FAO 2007).
MATERIALS AND METHODS
Avicennia marina (Forsk.) Vierh.) is a mangrove tree
species almost always found in major mangrove Sample collection and nutrient analysis
ecosystems (Tomlinson 1986). Local people use this plant's The materials used in this experiment consist of
stems and twigs for firewood, furniture, building materials,
boat balancing joints, and fishing net dyes (Armitage Avicennia marina leaves and fruit, Tithonia diversifolia
2002). These products are harvested on a small and large leaves, Gliricidia sepium leaves, Leucaena leucocephala
scale, contributing to local livelihoods and national exports. fruit, and leaves. A. marina leaves were taken from the
South Coast mangrove forest, Sebelas Tarusan Sub-district,
Avicennia marina leaves have a pointed shape at the tip Pesisir Selatan District, West Sumatra, Indonesia. T.
and are green at the front and grayish at the bottom with diversifolia, G. sepium, and L. leucocephala leaves were
about 5-11 cm. The flowers are small round with a collected from the experimental gardens of the Faculty of
diameter of about 0.4-0.5 cm and yellow to orange, while Agriculture, Andalas University, Padang, Indonesia.
the fruit is round with a pointed tip and smooth-haired
surface, green with a length of 1.5-2.5 cm and a width of

299Dasar Nutrisi Ruminansia (Edisi ke II)

JAMARUN et al. Chemical composition of mangrove leaves ( Avicennia marina) 5231

Leaves from these species have been traditionally used for formed from the addition of H2SO4) was taken, and one
feeding ruminants and therefore used as references for drop of Meyer reagent was added. A white mist
evaluating A. marina leaves' potency. characterized positive reactions. The flavonoid Test layer
of water as much as 2 ml from the preparation stage was
Avicennia marina leaf samples taken were the top 3-5 taken and put into a test tube. Then 1-2 grains of
leaves, such as tea leaves. The samples taken were dried at Magnesium was added, and three drops of HCl were added.
60 oC for 24 hours. The sample is grinded until smooth. All Positive samples contain flavonoids. If they form orange to
the leaf samples were oven-dried at 60oC for 24h. Concerning the phenolic test, a layer of water from the
Especially for A. marina leaves, the chemical composition, preparation stage was taken and put into a drip plate, then
rumen fluid profile, and nutrient digestibility were tested. added ferric chloride to each drip plate that has been
Proximate content was analyzed by standard methods, sampled. The formation of blue and purple characterizes
according to AOAC (2000). Neutral detergent fiber (NDF), the presence of phenolic compounds. A 2 ml layer of water
cellulose, and acid detergent fiber (ADF) were analyzed from the preparation stage was taken and put into a test
according to Van Soest et al. (1991). In-vitro rumen tube then shaken for the saponin test. Positive samples
incubation method followed the procedure of Tilley and contain saponins if they are formed permanently, which do
Terry (1963), macro and micro minerals using Inductively not disappear within 15 minutes. Steroid and triterpenoid
Coupled Plasma Optical Emission Spectroscopy (ICP- test was performed by taking the chloroform layer from the
OES) while phytochemical compounds by the Harborne preparation stage and put into a Pasteur pipette, which
(1998). All the analyses were carried out at the contains charcoal. The filtrate that comes out of Pasteur's
Biochemistry Laboratory of the Faculty of Pharmacy and pipette was inserted into three holes on the drip plate,
Water Laboratory of the Faculty of Engineering, Andalas adding one drop of anhydrous acetic acid and one drop of
University, Padang, Indonesia. All the data obtained were H2SO4. Positive samples containing steroid compounds
described descriptively. were shown in blue to purple, while positive samples
contain triterpenoid compounds if produced in red.
Phytochemical analysis
Phytochemical analysis was carried out by the Determination of mineral contents
Avicennia marina leaves and fruits, T. diversifolia
extraction method based on Franswort (1996). Before
phytochemical analysis, A. marina fruit and leaves, T. leaves, G. sepium leaves, L. leucocephala fruits and leaves
diversifolia leaves, G. sepium leaves, L. leucocephala fruit, were dried in an oven at 60°C for 24 hours. Then the
and leaves were ground into flour, put into a bottle, added sample was ground and filtered using a 20 mesh filter to
with 90% methanol solvent in a ratio of 1: 3 (w/v), obtain a powdered sample. One gram of powdered sample
macerated with solvent methanol 3x24 hours and every 24 was added with 2 ml of distilled water, then dried in the
hours the methanol solvent was replaced. The maceration furnace at 150 °C for 15 minutes. Then the sample was
results were then filtered using Whatman filter paper no. 42 cooled at room temperature. Dilute using aqua dest to a
so that the resulting filtrate. The filtrate was subjected to volume of 25 ml, and then the sample was filtered using 45
several phytochemical screening tests, i.e., alkaloid, mesh filter paper. The destruction results were analyzed in
flavonoid, phenolic, saponin, steroid, and triterpenoid tests. the mineral content of Fe, Zn, Mn, Cu, and Co using the
For the alkaloid test, the chloroform layer was added ten Inductively Coupled Plasma Optical Emission
drops of H2SO4 and shaken slowly, allowed to form an Spectroscopy (ICP-OES) tool.
acidic layer. A layer of acid (the part under the clear ring

Figure 1. Location of Avicennia marina leaf sampling in Sebelas Tarusan Sub-district, Pesisir Selatan District, West Sumatra, Indonesia

300 Dasar Nutrisi Ruminansia (Edisi ke II)

5232 21 (11): 5230-5236, November 2020

RESULT AND DISCUSSION Crude fiber is needed for ruminants to maintain the
development of rumen microbes. Crude fiber that is too
Chemical composition of Avicennia marina leaves low will interfere with the digestive system of ruminants.
Avicennia marina leaves contain 13.37% crude protein The A. marina crude fiber content (12.18%) is almost equal
to the minimum requirement of crude fiber content in feed
(CP) with 79% Total Digestible Nutrient (TDN) (Table 1). ingredients, which is 13% for cattle, according to
This value makes A. marina leaves included in the category Sudarmono and Sugeng (2008).
of high-quality forage. (Jamarun and Zain 2013) classify
forage quality in three categories based on CP and TDN The NDF content is closely related to feeding
content, namely low quality forage (CP <4%, TDN> 40%), consumption because all its components meet the rumen
medium quality forage (CP 5-10%, TDN 40-50%) and space and are slow to digest. The lower the NDF content,
high-quality forage (CP> 10%, TDN> 50%). High forage the more food can be consumed. ADF's content (cellulose,
CP and TDN are needed by livestock to optimize their lignin, silica) is an indicator of forage digestibility because
growth and production. Some CP in the rumen will be lignin's content is part of an indigestible fraction (Pazla et
overhauled into NH3 by proteolytic enzymes produced by al. 2020). NDF is always higher than ADF because ADF
rumen microbes. NH3 concentration is an important source does not contain hemicellulose. NRC (2001) suggests a
of N for rumen microbes and is used for microbial protein minimum of NDF in feed 21% with ADF 19%. The
synthesis. NH3 production is influenced by the amount of percentage of ADF and NDF content given to livestock
protein in feed ingredients (Pazla et al. 2018a). High TDN should be 25-45% ADF and 30-60% NDF from forage dry
illustrates that these leaves have a high digestibility, so matter (Anas 2010). The average value of lignin that
only a few nutrients come out as feces. livestock can tolerate is 7% (Goering and Van Soest 1970).
The NDF, ADF, and lignin values of these leaves are still
The high CP content in A. marina leaves is caused by within the tolerance range for ruminant animal feed.
soil organic matter (OM). (FAO 2007) In mangrove areas,
there is high organic matter in thick mud that lines the Table 1. Chemical composition of Avicennia marina leaves
surface of the land at low tide. Land influences nutrition,
plant growth, and development. Plants will grow and Chemical composition %
develop optimally if the soil conditions in which they live
fit the nutritional and nutrient requirements. According to Dry matter 89.19±0.07
Kennish (2000), mangrove roots can accumulate sediment Ash 7.17±0.09
and play a role in forming soil formations. Mangroves are Organic matter 92.83±0.11
suppliers of organic material to provide food for organisms Crude protein 13.37±0.23
that live in the surrounding waters. Sedimentation that Crude fiber 12.18±0.27
occurs in mangrove areas is different from other regions. Crude fat 3.18±0.39
Sources of sedimentation come from the land, sea, and NDF 45.99±0.41
mangrove areas in the form of deposited leaf deposits, ADF 35.95±0.43
twigs, and dead organisms that are collected so that this Cellulose 23.10±0.42
region is rich in organic and mineral materials such as N, P, Hemicellulose 10.03±0.67
K, Fe, and Mg (Nugroho et al. 2013). A. marina leaves' Lignin 7.34±0.72
crude protein value in this study was higher than reported TDN 79.00±0.98
by (Handayani 2013), 11.04%, and lower than (Ghosh et al.
2015), 15.14%. This variation in crude protein values can Table 2. Rumen fluid profile and nutrition in vitro digestibility
be caused by plant age, soil fertility, and the source (Jama from Avicennia marina leaves
et al. 2000).
Parameters Value
The high protein content of a feed ingredient will also
increase the value of organic matter. This is due to crude Rumen fluid profile 6.79±0.02
protein is part of organic material. Table 1 shows the pH 117.5±0.04
organic matter content of the leaves is also relatively high, VFA (mM) 16.88±0.51
at 92.83%. High organic matter will automatically reduce NH3 (mM)
the value of ash content. The higher the ash content, the 56.68±0.54
worse the quality of feed ingredients (Suparjo 2010). (SNI Nutrition digestibility (%) 63.74±0.67
2017) suggests that cattle's low ash content is 12 % DM 69.96±0.62
maximum, while poultry livestock is 8%. The low-crude fat OM 61.37±1.58
(CF) content in these leaves (3.18%) is advantageous in CP 57.44±0.96
ruminant animals. The high-fat content in feed ingredients CF 51.44±0.92
has been reported to be a cause of digestive and metabolic NDF 60.24±0.73
disorders in cattle (Atteh 2002). Preston and Leng (1987) ADF 62.03±1.04
supported this, and Palmquist and Jenkins (1980) stated Cellulose
that ruminant animal feed ingredients' standard fat content Hemicellulose
is below 5%.

301Dasar Nutrisi Ruminansia (Edisi ke II)

JAMARUN et al. Chemical composition of mangrove leaves ( Avicennia marina) 5233

Table 3. Mineral macrocontent of Avicennia marina, Thitonia diversifolia, Gliricidia sepium and Leucaena leucocephala

Mineral Avicennia marina Thitonia diversifolia Gliricidia sepium Leucaena leucocephala
content (%)
Leaf Fruit Leaf Leaf Leaf Fruit
Ca 0.21±0.007 0.25±0.014
Na 0.38±0.007 0.35±0.014 0.14±0.014 0.28±0.014 0.24±0.007
Mg 0.09±0 0.13±0.007
K 0.20±0.014 0.17±0.007 0.20±0.007 0.28±0.014 0.16±0.007 0.13±0
P 0.26±0.007 0.23±0.007
S 0.20±0.07 0.19±0.007 0.32±0.014 0.0063±0 0.16±0.007 0.15±0.007
Cl 0.0052±<0.001 0.92±0.021
0.48±0.021 0.41±0.014 0.89±0.014 0.32±0.014 0.27±0.014

0.51±0.014 0.47±0.014 0.42±0.014 0.23±0.014

0.01±0 0.0092±<0.001 0.0071±<0.001 0.0054±<0.001

1.03±0.021 0.99±0 0.96±0.014 0.85±0.021

Table 4. Mineral microcontent of Avicennia marina, Thitonia diversifolia, Gliricidia sepium and Leucaena leucocephala

Mineral content Avicennia marina Thitonia diversifolia Gliricidia sepium Leucaena leucocephala
(ppm)
Leaf Fruit Leaf Leaf Leaf Fruit
Fe 293±<0.001 228±<0.001
Zn 388±<0.001 293±<0.001 76±<0.001 56±<0.001 390±<0.001 328±<0.001
Mn 75±<0.001 56±<0.001
Cu 164±<0.001 135±<0.001 40±<0.001 43±<0.001 88±<0.001 75±<0.001

211±<0.001 139±<0.001 88±<0.001 75±<0.001

128±<0.001 107±<0.001 83±<0.001 53±<0.001

Table 5. Phytochemical composition test results of Avicennia marina, Tithonia diversifolia, Gliricidia sepium dan Leucaena
leucocephala

Test result

Parameters Avicennia Avicennia Tithonia Gliricidia Leucaena Leucaena
marina fruit marina leaves leucocephala leucocephala
Alkaloid diversifolia leaves sepium leaves
Flavonoid + - leaves fruit
Phenols - - -- - -
Saponin + + -- - -
Steroid - -
Triterpenoid + + -- - -
Tanin + + -+ - -
+ + ++ + +
++ + +
++ + +

Rumen fluid profile and nutrition digest ibility hemicellulose, which are the main components of forming
The pH value of the rumen fluid from A. marina leaves VFA. The high protein content of A. marina leaves also
contributed to the high VFA value. There is a positive
in this study was within the normal range for the growth correlation between high crude protein values and VFA
and development of rumen microbes, mostly bacteria values (Jamarun et al. 2017b; Jamarun et al. 2018).

(Table 2). The ideal pH for fiber digestion is 6.4-6.8 The concentration of NH3 in A. marina leaves in this
(France and Siddon 1993). The pH below 6.2 will reduce study was included in the amount of NH3 that supports
plant fiber digestibility because cellulolytic bacteria's rumen microbial growth, namely 6 mM-21 mM (Mc
activity is inhibited (Erdman 1988). A pH value above 7.1 Donald 2010). Paengkoum et al. (2006) stated that the
maximum NH3 concentration required for rumen microbes
can reduce the microbial population drastically so that the to digest feed was 3.57-14.28 mM. Rumen microbes use
energy generated from the rumen fermentation process is NH3 as a N for microbial protein synthesis source, and its
low (Van Soest 1982). value is also influenced by crude protein levels (Pazla et al.
2018a). The pH, VFA, and NH3 values of A. marina leaves
Volatile fatty acid (VFA) is a source of energy for the in this study were almost the same as other forages such as
growth and development of rumen microbes. The VFA T. diversifolia (6.78, 125.88mM, 22.48mM) and Elephant
grass (6.79, 87.53 mM 20, 41mM) (Jamarun et al. 2019).
value produced by A. marina leaves sufficient for rumen
microbes to grow and develop optimally. Mc Donald et al. Feed digestibility is a large amount of feed that
(2010) stated that the optimum VFA condition is 80-160 livestock can utilize to meet basic needs and production.
mM. The high value of the resulting VFA indicates that A. Based on Table 2 above, it can be seen that rumen
microbes can digest more than 50% of the nutrients from
marina leaves are a feed material with a high level of
fermentability, which is suitable as a source of forage for
ruminants. The low lignin content will make it easier for
enzymes from rumen microbes to penetrate cellulose and

302 Dasar Nutrisi Ruminansia (Edisi ke II)

5234 21 (11): 5230-5236, November 2020

these leaves; this is due to the low lignin content. Lignin in (2008) states that Mn functions as carbohydrate synthesis,
feed ingredients can reduce digestibility, as reported by mucopolysaccharide, and enzyme systems, such as
Jamarun et al. (2017a). Rumen microbes can digest food pyruvate carboxylase and arginine synthetase. In addition
substances in feed ingredients when the lignin content is to enzymatic reactions, Mn also functions for growth and
low. Imsya et al. (2013) stated that lignin in plant cell walls reproduction of livestock, Onwuka et al. (2001) which
limits the feed material's digestibility. Crude protein states that Mn's mineral content in goats ranges from 2.98-
content in feed ingredients will also affect the digestibility 13.9 mg/dl. Based on these data, it can be concluded that
level of a feed ingredient. The high protein content of A. the livestock reared with A. marina leaf-based feed does
marina leaves will provide more nitrogen for the growth of not experience Mn mineral deficiency because the Mn
rumen microbes. Profitable microbial growth will lead to content in the forage is sufficient. Nugroho's (2008)
better feed digestibility (Febrina et al. 2016). opinion states that Mn mineral deficiency rarely occurs
because Mn levels in the feed are enough for livestock
Macro and micro mineral contents needs.
The amount of macro minerals (Ca, Na, Mg, K, S, P,
Zinc (Zn) is the micro-mineral often deficient for rumen
and Cl) A. marina leaves is higher than that of A. marina microbial growth (Leng 1991). To maximize feed
fruit, T. diversifolia, G. sepium leaves, and L. leucocephala degradation in the rumen, the adequacy of Zn minerals is
leaves (Table 3). The high mineral content is because the critical, given the strategic role of Zn in increasing rumen
soil in the mangrove forest is rich in minerals and organic microbial growth and as an activator of many enzymes
matter. Nugroho et al. (2013) explained that the (Elihasridas et al. 2012). Mineral Zn can stimulate rumen
sedimentation in the mangrove area is different from other microbial growth and improve the appearance of livestock.
depositional environments. Sources of sediment in The Zn content in Indonesia's ruminant animal feed ranges
mangrove areas come from land and sea (allochthonous) from 20-38 mg/kg of dry ration material (Little 1986). This
and from the mangrove area itself (autochtonous) in the value is far below the need for rumen microbes, namely
form of heaps of fallen leaves, twigs, and dead organisms 130-220 mg/kg of dry ration material (Hungate 1966). Zn
deposited in the mangrove area and contain a lot of organic deficiency can interfere with rumen microbial metabolism
and mineral matter (N, P, K, Fe, and Mg). The and decrease enzyme activity. Therefore to achieve high
allochthonous sediment is deposited in mangroves through feed degradation and microbial growth in the rumen, Zn
sediment transport, where suspended particles are carried must be available in sufficient and balanced amounts. The
by tidal currents stored in the mangrove area. Because amount of Zn in A. marina leaves still in the range to meet
mangroves have a unique root system, they can reduce tidal the needs of rumen microbes.
currents in the mangrove area.
According to Darmono and Bahri (1989), the low Cu in
Macrominerals are needed by livestock to build body animal feed sources will adversely affect Fe intake, even
structures such as bones and teeth (Jamarun and Zain though the Fe content in the feed is adequate. It was
2013). P mineral is an important mineral to support the reported that low Cu content in forage is one of the causes
growth of rumen microbes digesting fiber (Suyitman et al. of anemia in livestock. According to Little (1986), several
2020). Sulfur minerals are needed by rumen microbes to types of grass or forage are used as sources of feed for
form amino acids that contain sulfur (Bal and Ozturk ruminants in Indonesia, especially on Sumatra, whose
2006). Mineral P and S can stimulate rumen microbial Cunya content is below average (low) limits. As reported
performance to improve feed digestibility (Pazla et al. by Prabowo et al. (1997) and Mathius (1988) from the
2018b). Mineral P, S, and Mg were able to increase rumen results of field examinations that are commonly used as the
VFA concentrations (Febrina et al. 2016). Minerals Ca, P, main feed for goats generally have Cu content below the
and Mg at normal levels in the rumen can increase rumen standard (critical) limit. The Cu content will be even lower
microbial activity in digesting cellulose and VFA (Adriani during the dry season. This results in animals that consume
and Mushawwir 2009). Na functions to increase appetite them, thus experiencing mineral deficiencies. Mc Dowell et
and maintain osmotic pressure (Jamarun and Zain 2013). A. al. (1992) states that Cu requirements are influenced by the
marina leaves' mineral content is still in the normal range levels of other mineral rations, which increases the need for
to help supply the mineral needs. According to Mc Dowell ruminants in the presence of high molybdenum (Mo)
et al. (1983) the range of normal values for mineral content levels. NRC (1989) recommends a Cu requirement figure
in animal feed for Ca is 0.17-1.53 %, Mg 0.05-0.25%, P of 10 mg/kg for ruminants. The mineral value of Cu in the
0.17 0.59%, K 0.50-0.70%, Na 0.01-0.06%, S 0.08-0.15%. leaves of the A. marina is sufficient for livestock needs.
The definition of Cu will cause bone disorders (paralysis),
Fe's mineral content in A. marina leaves relatively high joint swelling, bone fragility. Pigment deficiency in Cu -
compared to A. marina fruit, T. diversifolia, G. sepium, and deficient animals and humans. However, giving enough
L. leucocephala fruit, but L. leucocephala leaves have copper salt, especially to sheep, will cause accumulation in
slightly higher Fe (Table 4). Nugroho (2008) states that Fe the liver. Sheep are sensitive to 20-30 mg Cu/kg of Cu
content in grass is usually 100-200 ppm while in legume ration (Tillman et al. 1998).
200-300 ppm. According to Darmono (2007), mineral Fe is
used in the enzymatic metabolism of hemoglobin in the Phytochemical contents
livestock body. The phytochemical contents of the samples were varied

The minerals Zn, Mn, and Cu in A. marina leaves show (Table 5). Ruminant animals are more resistant to feed
the highest value than other forages in Table 3. Nugroho

303Dasar Nutrisi Ruminansia (Edisi ke II)

JAMARUN et al. Chemical composition of mangrove leaves ( Avicennia marina) 5235

ingredients that contain phytochemicals than poultry. This Bal MA, Ozturk D. 2006. Effects of sulfur-containing supplements on
is due to some phytochemicals that can be used to simplify
the process of feed metabolism. Tannins are ruminal fermentation and microbial protein synthesis. Res J Anim Vet
phytochemicals that function as by-pass protein agents.
This means that the protein from feed ingredients eaten by Sci 1: 33-36.
livestock will be protected from rumen bacteria's
degradation to enter the small intestine. This tannin can Bunting P, Ake R, Richard M, Lucas, Lisa MR, Lammert H, Nathan T,
only release its bonds with feed ingredients by enzymes in
the small intestine and low pH levels, while in the rumen, Andy H, Takuya I, Masanobu Sand C, Max F . 2018. The global
tannins are problematic in the rumen bacterial break and
normal rumen pH (Jamarun and Zain 2013). Tannin mangrove watch-a new 2010 global baseline of mangrove extent.
addition increased neutral detergent insoluble crude protein
(NDICP) and acid detergent insoluble CP (ADICP) Remote Sens 10 (10): 1669.
(Jayanegara et al. 2019). However, the levels should not be
excessive because if excessive phytochemicals can hurt Darmono, Bahri S. 1989. Defisiensi tembaga dan seng pada sapi di daerah
livestock productivity. Phytochemicals in feed ingredients
such as T. diversifolia, G. sepium, and L. leucocephala Kalimantan Selatan. Penyakit Hewan 21 (38): 128 -131. [Indonesian]
have been tested in livestock, apparently still in normal
conditions for consumption by ruminant animals and do not Darmono. 2007. Penyakit defiseinsi mineral pada ternak ruminansia dan
show a negative effect on livestock metabolic activities
(Arief et al. 2020; Pazla 2018; Ningrat et al. 2018). upaya pencegahannya. Jurnal Litbang Pertanian 26 (3): 104-108.
Jamarun et al. (2019) tested T. diversifolia by using up to a
100% level, still having a positive effect on the digestibility [Indonesian]
of dry matter and organic matter and the fermentability of
rumen fluids such as pH, NH3, and VFA, even better when Elihasridas. 2012. Respon suplementasi mineral zink (zn) terhadap
compared to elephant grass. Fasuyi et al. (2010) identified
many phytochemicals found in T. diversifolia such as kecernaan in-vitro ransum tongkol jagung amoniasi. Jurnal
phytates, alkaloids, saponins, and far more than A. marina
leaves. This study confirms that A. marina leaves are Peternakan 9 (2): 9-14. [Indonesian]
entirely safe for livestock consumption. In conclusion, the
research showed that A. marina leaves could be used as Erdman RA. 1988. Dietary buffering requirements of lactating dairy cows.
alternative feed ingredients for ruminant animals with CP
content of 13.37%, lignin 7.34%, rich in macro and micro A Review. J Dairy Sci 71: 3246-3246.
minerals, and containing phytochemical compounds such
as tannins, steroids, and triterpenoids. FAO. 2007. 1980-2005. Food and Agriculture

ACKNOWLEDGEMENTS Organization of the United Nations, Rome.

Andalas University funded this research by the research Fasuyi AO, Dairo FAS, Ibitayo FJ. 2010. Ensiling wild sunflower
contract No: T/12/UN.16.17/PP.KP-KPR1GB/LPPM/2019
the Fiscal Year 2019. (Tithonia diversifolia) leaves with sugar cane molasses. Livest Res

REFERENCES Rural Dev 22 (3): 42.

Adriani L, Mushawwir A. 2009. Level of blood glucose, lactose, and dairy Febrina D, Jamarun N, Zain M, Khasrad. 2016. The Effects of P, S and
cattle milk yield at a different level of macromineral
supplementation. J Indon Trop Anim Agric 34 (2): 88-95. Mg supplementation of oil palm fronds fermented by Phanerochayte

AOAC. 2000. Official methods of analysis. Association of analytical chrysosporium on rumen fluid characteristics and microbial protein
chemists, Arlington, Virginia, USA.
synthesis. Pak J Nutr 15 (3): 299-304.
Anas S. 2010. Kandungan NDF dan ADF silase campuran jerami ja gung
(I) dengan beberapa level daun gamal ( Gliricidia maculata). Balai France J, Siddons RC. 1993. Volatile fatty acids production . In: Forbes JM,
Pengkajian Teknologi Pertanian (BPTP), Gorontalo . [Indonesian]
France J (eds.). Quantitative Aspect of Ruminant Digestion and
Arief, Sowmen S, Rusdimansyah, Pazla R, Rizqan. 2020. Milk production
and quality of Etawa crossbreed dairy goat given Tithonia Metabolism. CAB Internasional, Wallingford, UK.
diversifolia, corn waste, and concentrate based palm kernel cake.
Biodiversitas 21 (9): 4004-4009. Franswort NR. 1996. Biological and phytochemical screenings of plant. J

Armitage D. 2002. Socio-institutional dynamics and the political ecology Pharm Sci 55: 225-265.
of mangrove observation Indonesia. Glob Environ Change 12: 203-
217. Ghosh S, Somraj C, Agniv N. 2015. Proximate composition of some

Atteh JO. 2002. Principles and practice of livestock feed manufacturing. mangrove leaves used As alternative fodders in Indian Sunderban
Adlek Printers, Ilorin, Niger.
Region. Intl J Livest Res 5 (11): 62-65.

Goering HK, Van Soest PJ. 1970. Forage fiber analysis. USDA Agric.

Handbook No. 379. USDA-ARS, Washington, DC.

Handayani S. 2013. Kandungan flavonoid kulit batang dan daun pohon

api-api (Avicennia marina (forks.)vierh.) sebagai senyawa aktif

antioksidan. [Thesis]. Bogor Agricultural University, Bogor.

[Indonesian]

Harborne AJ. 1998. Phytochemical Methods: A Guide to Modern

Techniques of Plant Analysis. Springer, Nederland.

Hungate RR. 1998. The rumen and its microbe. Department of

Bacteriology and Agriculture Experiment Station University of

California. Davis California Academy Pres s, London.

Imsya A, Laconi EB, Wiryawan KG, Widyastuti Y. 2013. In Vitro

digestibility of ration containing different levels of palm oil frond

fermented with Phanerochaete chrysosporium. Media Peternakan 36

(2): 131-136. [Indonesian]

Jama B, Palm CA, Buresh RJ. 2000. Tithonia diversifolia as a green

manure for soil fertility improvement in western Kenya: A

review. Agrofor Syst 49: 201-221.

Jamarun N, Zain M. 2013. Dasar nutrisi ruminansia. Jasa Surya Press,

Padang. [Indonesian]

Jamarun N, Zain M, Arief, Pazla R. 2017a. Effects of calcium, phosphorus

and manganese supplementation during oil palm frond fermentation

by Phanerochaete chrysosporium on laccase activity and in vitro

digestibility. Pak J Nutr 16 (3): 119-124.

Jamarun N, Zain M, Arief, Pazla R. 2017b. Effects of calcium (Ca),

phosphorus (p) and manganese (mn) supplementation during oil palm

frond fermentation by Phanerochaete chrysosporium on rumen fluid

characteristics and microbial protein synthesis. Pak J Nutr 16 (6):

393-399.

Jamarun N, Zain M, Arief, Pazla R. 2018. Populations of rumen microbes

and the in vitro digestibility of fermented oil palm fronds in

combination with tithonia (Tithonia diversifolia) and elephant grass

(Pennisetum purpureum). Pak J Nutr 17 (1): 39-45.

Jamarun N, Pazla R, Zain M, Arief. 2019. Comparison of in vitro

digestibility and rumen fluid characteristics between the tithonia

(Tithonia diversifolia) with elephant grass (Pennisetum Purpureum).

IOP Conf Ser: Earth Environ Sci 287 (1): 1-6.

Jayanegara A, Aldi Y, Lilis K. 2019. Reduction of Proteolysis of high

protein silage from moringa and Indigofera leaves by addition of

tannin extract. Vet World 12 (2): 211-217.

Kennish MJ. 2000. Estuary restoration and maintenance: the national

estuary program. CRC Press, Boca Raton, USA.

304 Dasar Nutrisi Ruminansia (Edisi ke II)

5236 21 (11): 5230-5236, November 2020

Kitamura S, Anwar C, Chaniago A, Baba S. 1997. Handbook of Pazla R, Jamarun N, Zain M, Arief. 2018a. Microbial Protein synthesis

mangroves in Indonesia; Bali & Lombok. The development of and fermentability of fermented oil palm fronds by Phanerochaete
sustainable mangrove management project, Ministry of Forest chrysosporium in combination with tithonia ( Tithonia diversifolia)
Indonesia and Japan International Cooperation Agency, Denpasar. and elephant grass ( Pennisetum purpureum). Pak J Nutr 17 (10): 462-

[Indonesian] 70.
Leng RA. 1991. Feeding strategies for improving milk production of dairy Pazla R, Zain M, Ryanto I, Dona A. 2018b. Supplementation of minerals

animals managed by small farmers in the tropics. Feeding dairy cows (phosphorus and sulfur) and Saccharomyces cerevisiae in a sheep diet
in the tropics. (eds) Speedy A, Sansoucy R. Proceedings of the FAO based on a cocoa by-product. Pak J Nutr 17 (7): 329-335.
Expert Consultation held in Bangkok, Thaila nd, 82. Pazla R, Jamarun N, Agustin F, Zain M, Arief, Cahyani NO. 2020. Effects
Little DA. 1986. The Mineral Content of ruminant feeds and potential for of supplementation with phosphorus, calcium and manganese during

mineral supplementation in southeast Asia with particular reference to oil palm frond fermentation by Phanerochaete chrysosporium on
Indonesia. In: Dixon RM (ed). Proc. of the Fifth Annual Workshop of ligninase enzyme activity. Biodiversitas 21 (5): 1833-1838.
the Australian-Asia Ruminant Feeding System Utilizing Fibrous Prabowo A, Djajanegara A, Diwyanto K. 1997. Nutrisi mineral pada

Agricultural Residues. 1985. Int. Dev. Prog. of Austr. Univ. and ternak ruminansia. Jurnal Litbang Pertanian 16 (2): 53-64.
Colleges Limited (IDP), Canberra, Australia. [Indonesian]
Mathius IW. 1988. Jenis dan nilai gizi hijau an makanan domba dan Preston TR, Leng RA. 1987. Matching ruminant production systems with

kambing di pedesaan Jawa Barat. Dalam : Prosiding Pertemuan Ilmiah available resources in the t ropics and sub-tropics. The Technical
Ruminansia. Jilid 2, Ruminansia Kecil. Cisarua, Bogor. [Indonesian] Centre for Agricultural and Rural Co -operation (CTA). Wageningen,
Mc. Donald P, Edwards RA, Greenhalgh JFD, Morgan CA. 2010. Animal Netherlands.
Nutrition. 7th ed. Longman, New York. Rahardian A, Lilik BP, Yudi S, Ketut W. 2019. Tinjauan historis data dan
Mc Dowell LR, Conrad, JH, Ellis GL Loosli JK. 1983. Minerals for informasi luas mangrove Indonesia. Media Konservasi 24 (2): 163-78.
grazing ruminants in tropical regions. Universitas of Florida and The Richards DR, Friess DA. 2016. Rates and drivers of mangrove

Agency for International Development, USA. deforestation in Southeast Asia, 2000-2012. Proc Natl Acad Sci USA
Ningrat RWS, Zain M, Erpomen, Suryani H. 2018. Effects of 113 (2): 344-349.
Standar Nasional Indonesia (SNI). 2017. Pakan konsentrat -
supplementation of different sources of tannins on nutrient
Potong. Badan Standarisasi Nasional , Jakarta. [Indonesian]
digestibility, methane production and daily weight gain of beef cattle Sudarmono AS, Sugeng YB. 2008. Edisi Revisi Sapi Potong. Penebar
fed an ammoniated oil palm frond based diet. Int l J Zool Res 14: 8-
13. Swadaya, Jakarta. [Indonesian]

NRC. 1989. Nutrient Requirement of Dairy Cattle. 7th Edition. Nationa l Suparjo. 2010. Diktat Laboratorium Makanan Ternak. Fakultas
Academic of Science, Washington DC., USA. Peternakan Universitas Jambi, Jambi. [Indonesian]

NRC. 2001. Nutrient requirements of beef cattle: Seventh Revised Suyitman, Warly L, Rahmat A, Pazla R. 2020. Digestibility and

Edition: Update 2000. Subcommittee on Beef Cattle Nutrition. performance of beef cattle fed ammoniated palm leaves and fronds
Committee on Animal Nutrition. National Research Council , USA. supplemented with minerals, cassava leaf meal and their
Nugroho P. 2008. Agribisnis ternak ruminansia Jilid 1 Untuk SMK. combinations. Adv Anim Vet Sci 8 (9): 991-996.
Direktorat Pembinaan Sekolah Menengah Kejuruan, Direktorat Tilley JMA, Terry RA. 1963. A two-stage technique for the in vitro
Jenderal Manajemen Pendidikan Dasar dan Menengah, Departemen digestion of forage crops. J Brit Grassl Soc 18 (2):104-111.
Pendidikan Nasional, Jakarta. [Indonesian] Tillman AD, Hartadi H, Reksohadiprojo S, Prawirokusumo S,

Nugroho RA, Sugeng W, Rudhi P. 2013.Studi kandungan bahan organik Lebdosukodjo S. 1998. Ilmu Makanan Ternak Dasar. Gadjah Mada
dan mineral (N, P, K, Fe dan Mg) sedimen di kawasan mangrove University Press, Yogyakarta. [Indonesian]
Desa Bedono, Kecamatan Sayung. J Mar Res 2: 62-70. Tomlinson PB. 1986. The Botany of Mangroves. Cambridge University

Onwuka SK, Avwioro OG, Akpan MO, Ahmed Y. 2001. Distribution of Press, Cambridge.
cobalt, manganese, and iron in the skin and hair of West African United Nations Environment Program. 2014. United nations environment
dwarf sheep and goat in Nigeria. Afr J Biomed Res 4: 151 -154.
program world conservation monitoring center. The Importance of
Paengkoum P, Liang JB, Jelan ZA, Basery M. 2006. Utilization of steam-
treated oil palm fronds in growing Saanen goats supplementation with Mangroves to People: A Call to Action .
energy and urea. Asian-Aust J Anim Sci 19 (11): 1623-1631. http://newsroom. unfccc.int/es/el -papel -de-la -naturaleza/la -onu -al erta -
de-la -rapida -destruccion-de-los-manglares/.
Palmquist DL, Jenkins TC. 1980. Fat in lactation ration: a review. J Dai ry Van Soest PJ. 1982. Nutritional ecology of the ruminant metabolism
Sci 63: 1-14. chemistry and forage and plant fiber. Cornell University, Oregon,
USA.
Pazla R. 2018. Pemanfaatan Pelepah Sawit dan Titonia (Tithonia
Van Soest PJ, Robertson JB, Lewis BA. 1991. Methods for dietary fiber,
diversifolia) dalam Ransum Kambing Peranakan Etawa untuk neutral detergent fiber, and nonstar ch polysaccharides in relation to
Menunjang Swasembada Susu 2020. [Disertasi]. Universitas Andalas, animal nutrition. J Dairy Sci 74 (10): 3583-3597.
Padang. [Indonesian]

305Dasar Nutrisi Ruminansia (Edisi ke II)

306 Dasar Nutrisi Ruminansia (Edisi ke II)

307Dasar Nutrisi Ruminansia (Edisi ke II)

308 Dasar Nutrisi Ruminansia (Edisi ke II)

309Dasar Nutrisi Ruminansia (Edisi ke II)

310 Dasar Nutrisi Ruminansia (Edisi ke II)

311Dasar Nutrisi Ruminansia (Edisi ke II)

312 Dasar Nutrisi Ruminansia (Edisi ke II)

313Dasar Nutrisi Ruminansia (Edisi ke II)

Volume 22, Number 9, September 2021 ISSN: 1412-033X
Pages: 3936-3942 E-ISSN: 2085-4722
DOI: 10.13057/biodiv/d220940

Quality evaluation of tithonia (Tithonia diversifolia) with fermentation
using Lactobacillus plantarum and Aspergillus ficuum at different
incubation times

RONI PAZLA , NOVIRMAN JAMARUN1, , MARDIATI ZAIN1, GUSRI YANTI2, RIKI HISKIA CHANDRA3

1Department of Nutritional Science and Feed Technology, Faculty of Animal Husbandry, Universitas Andalas. Jl. Unand, Limau Manis Campus, Padang
25163, West Sumatra, Indonesia. Tel./fax.: +62-751-71464, email: [email protected], email: [email protected]

2Doctoral Program, Faculty of Animal Husbandry, Universitas Andalas. Jl. Unand, Limau Manis Campus, Padang 25163, West Sumatra, Indonesia
3Graduate Program, Faculty of Animal Husbandry, Universitas Andalas. Jl. Unand, Limau Manis Campus, Padang 25163, West Sumatra, Indonesia

Manuscript received: 19 July 2021. Revision accepted: 26 August 2021.

Abstract. Pazla R, Jamarun N, Zain M, Yanti G, Chandra RH. 2021. Quality evaluation of tithonia (Tithonia diversifolia) with
fermentation using Lactobacillus plantarum and Aspergillus ficuum at different incubation times. Biodiversitas 22: 3936-3942. This
research aimed to evaluate the nutritional quality of tithonia (Tithonia diversifolia) fermented using Lactobacillus plantarum and
Aspergillus ficuum with different incubation times on nutritional content, digestibility, phytase enzyme activity, and characteristics of
rumen fluid. The research used an experimental method with a factorial completely randomized design to evaluate the nutritional
content and phytase enzyme activity after fermentation (stage 1). A randomized block design was used to evaluate in vitro digestibility
and rumen fluid characteristics (stage 2). For factorial completely randomized design, factor A is the type of microbe (L. plantarum and
A. ficuum), then factor B is the incubation time (3,5,7 days). Parameters observed were the nutritional content of dry matter (DM),
organic matter (OM), crude protein (CP), crude fiber (CF), and phytase enzyme activity. For the Randomized Block Design, the
research treatments were A = A. ficuum + 5 days of incubation, B = A. ficuum + 7 days of incubation, C = L. plantarum + 3 days of
incubation, D = L. plantarum + 5 days of incubation. Parameters measured were the digestibility of dry matter(DMD), organic matter
(OMD), crude protein (CPD), crude fiber (CFD), rumen pH, VFA production, and NH3 rumen fluid. The results showed that there was
an interaction between the type of microbe and incubation time . The treatment had a significantly different effect (P<0.05) on the
content of OM, CP, CF and phytase enzyme activity, but no interaction with the content of DM treatment. In the digestibility, the results
showed that the effect was not significantly different (P>0.05) on DMD, OMD, CPD but had a significant effect (P <0.05) on CFD. The
treatments also had no significant effect (P>0.05) on VFA but were significantly different (P<0.05) on NH3. From this study, it can be
concluded that fermented tithonia using A. ficuum with an incubation period of 7 days could improve the quality of tithonia. It can be
seen from the content of CP (31.02%), CF (16.52%), phytase enzyme activity (37.36 U/ml), DMD (66.86%), OMD (67.36%), CFD
(81.01%), CPD (70.37%), VFA production 135 mM and NH3 concentration 14.31 mg/100 ml, and pH value 6.72 which is suitable for rumen
microbial growth.

Keywords: Aspergillus ficuum, digestibility, enzyme activity, fermentation, L. plantarum, Tithonia diversifolia

INTRODUCTION 4.10 tons/ha-10.20 tons/ha dry biomass production or
24.00-46.80 tons/ha/year fresh production. Tithonia as
Forage is the primary source of feed for ruminants. animal feed has not been widely used; this is because there
Forage feed serves to meet the needs of livestock both for is not much information about the use of these plants as
basic life, growth, reproduction, and production. Ruminant animal feed. Also, tithonia is often considered a weed so
livestock that experiences a shortage of forage feed that it is thrown away, even though it has a fairly good
ingredients will be stunted in the growth process. Various nutritional content. The nutritional content of whole
efforts to increase livestock production to meet animal tithonia (leaves + stems) are dry matter (DM), (25.57%),
protein sources' needs will be challenging if the availability organic matter (OM), (84.01%), crude protein (CP),
of forage is not proportional to the needs and existing (22.98%), and crude fiber (CF), (18.17%) (Jamarun et al.
livestock population. One of the efforts to overcome the 2017a). Therefore, tithonia has good potential to be used
shortage of forage feed ingredients is to look for alternative and developed as forage feed because it has good
feeds with high nutritional content, high production, and nutritional content and high productivity.
easy to adapt. One type of plant that can be used is tithonia
(Tithonia diversifolia). The obstacle to using tithonia for animal feed is anti-
nutritional substances such as phytic acid, tannins,
Tithonia can grow along roads, lakesides, on the edge saponins, oxalate, alkaloids, and flavonoids (Aye 2016).
of rice fields, and vacant land not used by the community These limiting factors, both substances directly contained
(Arief et al. 2019; Arief et al. 2020). This plant has large in feed ingredients or through metabolic products in
roots, many branches, a soft trunk and overgrows so that in livestock, can interfere with the use of feed. Also, it can
a short time, it can form a dense bush. Hafis (2019) stated affect the health and production of animals through
that tithonia plants harvested six times a year can produce mechanisms of decreasing nutrient intake, digestive and

314 Dasar Nutrisi Ruminansia (Edisi ke II)

PAZLA et al. Nutritional evaluation of Tithonia diversifolia using fermentation technology 3937

absorption disorders and causing other adverse side effects. MATERIALS AND METHODS
Oluwasola and Dairo (2016) stated that the most
antinutrient content in tithonia was phytic acid, 79.2 Sample collection and nutrient analysis
mg/100gr. The high phytic acid content in tithonia causes a Tithonia was taken in the Koto Lalang agricultural land
bitter taste, so it is not liked by livestock. Phytic acid in a
material can also interfere with mineral absorption because area, Kuranji, Padang City, West Sumatra Province. For the
phytic acid has chelating agent properties that can bind fermentation process, In-vitro treatment and chemical
minerals so that the biological availability of these minerals analysis were carried out at the Feed Industry and
decreases. Phytic acid can also bind to proteins and Technology Laboratory, Ruminant Nutrition Laboratory,
carbohydrates so that the digestion and absorption of these and Biotechnology Laboratory from Andalas University,
nutrients are disturbed (Selle et al. 2021). Padang, Indonesia. Proximate analysis was carried out
based on the (AOAC 2010) method. In vitro test using the
Various processing methods can be used to reduce anti- method of Tilley and Terry (1963). Phytase enzyme
nutritional substances in feed ingredients, one of which is activity follows the Alltech method with some
through fermentation technology. Fermentation is the modifications (Shieh and Ware 1968).
process of breaking down organic compounds into simple
ones involving microorganisms. The fermentation process Rejuvenation of Aspergillus ficuum and Lactobacillus
can increase food substances such as protein and energy plantarum
and break down complex components into simple components
(Jamarun et al. 2017b). Fermentation can also increase the The rejuvenation of A. ficuum was carried out on potato
nutritional value of low-quality ingredients and function in dextrose agar sloping media, incubated at 30 °C for seven
preserving feed ingredients, and one way to remove days. In contrast, L. plantarum was grown on MRS Broth
antinutrients or toxins contained in feed ingredients. media at 37°C for 48 hours. The MRS Broth media was
weighed as much as 5.52 grams and then added 100 ml of
Microbes that can be used for fermentation in tithonia distilled water, heated until it was clear on a hot plate.
are Lactobacillus plantarum bacteria and Aspergillus Cover with cotton and aluminum foil, then sterilize using
ficuum mold. These microbes produce a phytase enzyme an autoclave at 121°C for 30 minutes. Then cool and
(Myo-inositol hexakisphosphate phosphohydrolase) which inoculate with 1 ml of the bacterial parent. Incubate in an
can hydrolyze phytic acid (myo-inositol hexakisphosphate) incubator at 37 °C.
into inorganic monophosphate. Also, it can hydrolyze low
myo-inositol and some into free Myo-inositol. Therefore, Titonia fermentation process
the nutrients bound by phytic acid can be utilized. One hundred grams of tithonia flour was put into a
Sumenger et al. (2013) reported that L. plantarum could
produce high intracellular and extracellular phytase plastic bag and added with distilled water until the water
enzymes. Susana et al. (2000) also said that A. ficuum had content reached 60% and then homogenized. Sterilize in an
high activity and production of phytase enzymes from 60 autoclave for 30 minutes at a temperature of 121°C. After
isolates collected. Fermentation applications of A. ficuum the medium has cooled, 10% of the sample weight has been
and L. plantarum to reduce the phytic acid content in inoculated with microbes and 5 ml for L. plantarum, then
tithonia have not been carried out. The selection of these stored in a sterile room. Then, samples were harvested after
microbes in this study is because these microbes are 3,5 and 7 days and weighed fresh, then in the oven to dry at
relatively safe, not pathogenic, and have been widely 60°C. If it is dry, the medium is stored and ready for the
applied in fermentation. next stage, namely proximate analysis and In-vitro
digestibility testing.
Fermentation time is also one of the factors that must be
considered in making fermented feed. The fermentation Research design
that is too short resulted in limited opportunities for Stage 1; the research method used to evaluate the
microorganisms to grow. The substrate components that
can be remodelled into cell mass will also be small; for this chemical composition of post-fermentation tithonia, is an
reason, proper fermentation time is needed so that experimental method with a randomized completely design
microorganisms have more opportunities to grow and with a factorial pattern of 2 x 3 with three replications. The
reproduce. The ability of microorganisms to grow and parameters measured were dry matter, organic matter,
develop will affect the quality of a feed ingredient crude protein, crude fiber, and phytase enzyme activity.
physically, chemically, and biologically. Therefore, by The research treatment is as follows:
knowing the adequate time for the development of
microorganisms, we can get the best fermentation results in The first factor is the addition of microorganisms (A):
terms of quality. This study aimed to evaluate the quality of A1 = Tithonia + L. plantarum
fermented tithonia using L. plantarum and A. ficuum with A2 = Tithonia + A. ficuum
different fermentation times in vitro. The second factor is the length of fermentation (B):
B1 = 3 days
B2 = 5 days
B3 = 7 days
Stage 2; the experimental design used in the in vitro test
was a randomized block design with four treatment
combinations and three groups (rumen fluid collection) as
replication. The treatment in this second stage was based

315Dasar Nutrisi Ruminansia (Edisi ke II)

3938 22 (9): 3936-3942, September 2021

on the results of the best nutritional value (4 best Organic matter content of fermented tithonia
treatments) in stage one. The parameters measured were The organic matter content of fermented tithonia with
dry matter digestibility (DMD), organic matter digestibility
(OMD), crude protein digestibility (CPD), crude fiber different microbes and incubation time for each treatment
digestibility (CFD), total VFA concentration, NH3 is presented in Table 1.
concentration, and rumen fluid pH.
The analysis of variance showed an interaction
The treatments tested are: (P<0.05) between different types of microbes and
A: Fermented tithonia with A. ficuum for five days incubation time on OM content of fermented tithonia.
B: Fermented tithonia with A. ficuum for seven days Factor A (type of microbe) had an insignificant effect
C: Fermented tithonia with L. plantarum for three days (P>0.05), but factor B (incubation time) has a significant
D: Fermented tithonia with L. plantarum for five days impact (P<0.05) on the OM content of fermented tithonia.
The results of the DMRT test showed that the OM content
Data were analyzed using analysis of variance in the A2B1 treatment was significantly (P<0.05) higher
(ANOVA) according to (Steel and Torrie 2002). If the than the A1B1, A1B2, A1B3, and A2B3 treatments. The
study of variance results shows a significant effect, further high and low content of OM in the treatment is also made
tests are carried out with Duncan's Multiple Range Test. possible by microbial activity in the fermentation process,
which causes the breakdown of the substrate content,
RESULT AND DISCUSSION making it easier for existing microorganisms to digest
organic matter. The fermentation of organic matter releases
Dry matter content of fermented tithonia fermented products in the form of sugar, alcohol, and
The dry matter content of fermented tithonia with amino acids caused by the activity of micro-services so that
changes occur that affect the nutritional value. It is
different microbes and incubation time for each treatment following the opinion of Putra et al. (2019), which states
is presented in Table 1. that the fermentation process carried out by
microorganisms will cause changes that affect the
The analysis of variance showed no interaction between nutritional value in which carbohydrates are converted into
the treatment of different types of microbes and incubation alcohol, organic acids, water, and CO2.
time on the DM content of fermented tithonia. However,
every microbial treatment and incubation time showed a The decrease in OM content is due to the nutrients that
significant effect (P<0.05) on DM content. DMRT test have been utilized and remodelled by microbes. Microbial
results showed that the DM content of the A1 (L. growth is closely related to the length of fermentation,
plantarum) treatment showed a significant impact (P<0.05) where microbial development cycles start from the growth
with the A2 (A. ficuum) treatment. Incubation time in phase to the death phase. Mirnawati et al. (2013) added that
treatment B1 (3 days incubation) showed significantly the longer the fermentation time, the more food substances
different results (P<0.05) with treatment B2 (5 days were overhauled.
incubation) and treatment B3 (7 days incubation), while
treatment B2 (5 days incubation) showed similar results, Table 1. The phytochemical characteristic of fermented tithonia
not significantly different (P>0.05) with treatment B3 (7 with different types of microbes and incubation time
days incubation).
Factor A Factor B (incubation time) Average
The difference DM content in treatments A1 and A2 (Types of microbes) B1 B2 B3
was associated with the type and ability of each different
microbe in fermenting a material. Bacteria have a faster Dry matter
growth at optimum conditions when compared to molds, so A1 90.17 89.86 89.49 89.84 B
that the ability to digest substrate is also more
extraordinary. Bacteria as inoculum in the fermentation A2 91.76 91.26 90.12 91.05 A
process takes less time than molds; bacteria have a more
straightforward cell structure. So most bacteria have a Average 90.96a 90.56 b 89.8 b 90.44
shorter generation time when compared to molds whose
cell structure is more complicated and the generation time Organic matter
is quite long (Saylor and Casale 2020).
A1 87.78aA 87.51abA 87.04bA 87.44
The high DM content in treatments B1 was influenced
by the short fermentation time. According to Mirnawati et A2 88.40aB 87.91aA 86.67bA 87.66
al. (2010), the short fermentation time resulted in the 88..09a 87.71 a 86.86 b 87.55
substrate decomposition process not being optimal, so that Average
the water content was low, and DM was still high. During
the fermentation process, the substrate undergoes a Crude protein
decomposition process that causes changes in water
content. Changes in DM occur due to evaporation, A1 29.01aA 27.56bB 24.30cB 26.96 B
substrate hydrolysis, or metabolic water production.
A2 27.25cB 29.14bA 31.02aA 29.14 A

Average 28.13 28.35 27.66 28.05

Crude fiber
A1 17.95aB 17.37abA 17.18bA 17.50

A2 18.84aA 16.75bA 16.52bA 17.37

Average 18.40a 17.06b 16.85b 17.44

Phytase activity

A1 9.12bB 11.71aB 6.68cB 9.17B

A2 28.37cA 36.12bA 37.46aA 33.98A

Average 18.74c 23.91a 22.07b 21.58

Note: Values with different superscripts in the row (lower case)

and columns (capital letters) are significant (P<0.05). A1= L.

plantarum, A2= A. ficuum, B1= 3 days, B2= 5 days, B3= 5 days

316 Dasar Nutrisi Ruminansia (Edisi ke II)

PAZLA et al. Nutritional evaluation of Tithonia diversifolia using fermentation technology 3939

The crude protein content of fermented tithonia with A. ficuum could reduce CF content by up to 10.29%.
The crude protein content of fermented tithonia with The content of CF is still high due to the short fermentation
time; the microbes have not thrived and evenly, so the
different microbes and incubation time for each treatment enzymes produced to degrade CF have not worked
is presented in Table 1. optimally in reducing the CF content.

The analysis of variance showed an interaction Phytase enzyme activity of fermented tithonia
(P<0.05) between different types of microbes and The phytase enzyme activity of fermented tithonia with
incubation time on OM content of fermented tithonia.
Factor A (type of microbe) had an insignificant effect different microbes and incubation time for each treatment
(P>0.05), but factor B (incubation time) has a significant is presented in Table 1.
impact (P<0.05) on the OM content of fermented tithonia.
The results of the DMRT test showed that the OM content The analysis of variance showed that there was an
in the A2B1 treatment was significantly (P<0.05) higher interaction (P<0.05) between the type of microbe and the
than the A1B1, A1B2, A1B3, and A2B3 treatments. The length of incubation time on the activity of the fermented
high and low content of OM in the treatment is also made tithonia phytase enzyme. Factor A (type of microbe) and
possible by microbial activity in the fermentation process, factor B (incubation time) also had a significant effect
which causes the breakdown of the substrate content, (P<0.05) on the activity of the fermented tithonia phytase
making it easier for existing microorganisms to digest enzyme. The results of the DMRT test showed that the
organic matter. The fermentation of organic matter releases movement of the phytase enzyme in the A2B3 treatment
fermented products in the form of sugar, alcohol, and was significantly (P<0.05) higher than the other treatments.
amino acids caused by the activity of micro-services so that The increased activity of the phytase enzyme is due to A.
changes occur that affect the nutritional value. It is ficuum reaching the optimum conditions for its growth, the
following the opinion of Putra et al. (2019), which states better the development of the mold, the more enzymes
that the fermentation process carried out by produced, so that the enzyme activity will be higher. The
microorganisms will cause changes that affect the lowest enzyme activity was found in the A1B3 treatment,
nutritional value in which carbohydrates are converted into this was due to cell death which resulted in decreased
alcohol, organic acids, water, and CO2. enzyme activity. After reaching the optimum point,
microbial growth will slowly decrease because it begins to
The decrease in OM content is due to the nutrients that enter the death phase so that the activity of the phytase
have been utilized and remodelled by microbes. Microbial enzyme begins to fall.
growth is closely related to the length of fermentation,
where microbial development cycles start from the growth According to Maryanti (2015), the growth phase begins
phase to the death phase. Mirnawati et al. (2013) added that with the lag phase (adaptation phase), where microbes
the longer the fermentation time, the more food substances adjust to changes in the media and their environment. The
were overhauled. phase occurs shortly after inoculation, where the cells have
not experienced growth, and the number is still relatively
The crude fiber content of fermented tithonia constant. Next is the log phase (growth phase). As the
The crude fiber content of fermented tithonia with name implies, this phase is a phase of cell growth
characterized by a significant increase in the number of
different types of microbes and incubation time for each cells because the cell division process occurs optimally.
treatment is presented in Table 1. This phase is the best in determining the optimal time of
inoculation of a cell. The third phase is the stationary
The analysis of variance showed an interaction phase, in which cells will no longer grow and remain
(P<0.05) between the types of microbes and the length of relatively constant. The condition is due to reduced
incubation time on the CF content of fermented tithonia. nutrients and increased waste in the growth medium. This
Factor A (type of microbe) had an insignificant effect phase continues until it enters the death phase, which
(P>0.05), but factor B (incubation time) had a significant means the number of cells decreases drastically. Cells
effect (P<0.05) on the CF content of fermented tithonia. begin to die because the concentration of nutrients is
The results of the DMRT test showed that the CF content deficient and causes cell growth to be inhibited.
in the A2B1 treatment was significantly (P<0.05) higher
than the A1B1, A1B2, A1B3, A2B2, A2B3 treatments. Nutritional digestibility of fermented tithonia
Table 1 shows that the lowest decrease in CF content was Digestibility of dry matter, organic matter, crude
found in the A2B3 treatment (A. ficuum and incubation
time of 7 days) of 16.52%. The low content of CF in the protein, and crude fiber from fermented tithonia are
A2B3 treatment was due to the A. ficuum producing presented in Table 2.
cellulase enzymes. The cellulase enzymes could work
optimally in reducing crude fiber content. Cellulase The analysis of variance showed that fermented tithonia
enzyme breaks beta-1,4 glycosidic bonds in cellulose, with different types of microbes and incubation time had
cyclodextrin, cellobiose, and other cellulose derivatives no significant effect (P>0.05) on DMD of fermented
(Mingardon et al. 2011; Alami et al. 2017). tithonia. This is because the CF content of the treatment is
relatively the same. Digestibility is closely related to the
The longer the fermentation process, the lower the CF chemical composition of the material, especially the CF
content; this is due to the increased opportunity for A. content. CF content in treatments A, B, C and D
ficuum to degrade CF from the treatment substrate. respectively 16.75; 16.52; 17.95 and 17.37%. The DMD
Maulana (2019) reported that soy milk dregs fermented

317Dasar Nutrisi Ruminansia (Edisi ke II)

3940 22 (9): 3936-3942, September 2021

value in each treatment ranged from 62.21 to 66.86. DMD thicker and resistant to fiber-digesting microbes. It can
is higher than the study of Susanti et al. (2020), where result in a decrease in the digestibility of the feed
DMD in the combination of fermented tithonia and ingredients. It can be seen from the lowest CFD found in
sugarcane shoots was 59.15%, and Jamarun et al. (2019), treatment C (71.52%), with a crude fiber content of 17.95%
which obtained DMD in fresh tithonia of 58.56%. higher than treatments A, B, and D.

The analysis of variance showed that tithonia Rumen fluid characteristics of fermented tithonia
fermentation with different types of microbes and Rumen fluid characteristics from fermented tithonia are
incubation time gave no significant effect (P>0.05) on the
OMD. The value of OMD in this study ranged from 66.03 presented in Table 3.
to 67.36%. This value is not much different from the DMD The analysis of variance showed that tithonia
of fermented tithonia. The difference is not the OMD. After
all, the OMD is very closely related to the DMD because fermentation with different types of microbes and
some dry matter consists of organic matter. The OMD incubation time gave no significant effect (P>0.05) on the
pattern followed the DMD pattern (Pazla et al. 2018a; Arief pH of the rumen fluid. The pH value of the rumen fluid in
et al. 2021; Jamarun et al. 2021). The value of OMD in this this study ranged from 6.72 to 6.85, and the pH value was
study was higher than the study of Jamarun et al. (2019), still in a condition that was quite optimal for rumen
which obtained a digestibility value of organic matter of microbial growth. Jamarun and Zain (2013) stated that the
55.46% in tithonia without fermentation. optimal rumen pH for digestive activity in the rumen
ranged from 6.0 to 7.0. Rumen fluid pH less than 6.0 or
The analysis of variance showed that fermented tithonia above 7.0 can inhibit rumen microbial activity so that the
with different types of microbes and incubation time had ability to degrade feed decreases.
no significant effect (P>0.05) on the CPD of fermented
tithonia. These results illustrate that the ability of rumen Changes in rumen pH related to the production of
microbes to utilize protein is relatively the same. Besides, volatile fatty acid (VFA) produced in Table 3 show that the
digestibility is closely related to the chemical composition higher the production of VFA produced, the lower the
of the ingredients. If the design in feed ingredients is rumen pH. VFAs contain organic acids such as acetic,
relatively the same, the digestibility produced will also be propionate, butyrate, which make the atmosphere in the
pretty similar. The CPD material is also directly rumen acidic. Bhatia and Yang (2017) stated that the higher
proportional to the crude protein content of the material in the value of VFA, the more other organic acids (acetic,
the feed. propionic, butyric, isobutyrate, and isovalerate) were
produced the pH of the rumen fluid would below.
In addition, another factor that caused the CPD not However, based on the results, the rumen pH was not
significantly different in the treatment was the crude fiber significantly different from each treatment A, B, C, and D.
content of the material between treatments which was It was influenced by the provision of Mc Dougall buffer
relatively the same. CPD values in this study ranged from (artificial saliva), which played a role in maintaining pH.
67.78 to 70.38. This result is higher than the study of According to Van Soest et al. (1991), the condition of
Susanti et al. (2020), which obtained a CPD value of rumen pH remains constant due to the buffering capacity
55.45% in the combination of sugarcane top and fermented from saliva because it contains a lot of bicarbonate and
tithonia. phosphate and the absorption system of VFA through the
rumen wall.
The analysis of variance showed that fermented tithonia
with different types of microbes and incubation time had a The degree of acidity in the rumen is affected by the
very significant effect (P<0.05) on the CFD. DMRT further type of feed given but generally remains at a constant pH.
test results showed that treatment A was significantly Stable rumen pH in this study created a familiar
different (P < 0.05) with treatment B, C, and D. Treatment atmosphere in the rumen and is suitable for rumen
B was quite different (P < 0.05) with treatment C and D. microbes in carrying out their activities. Jamarun et al.
Treatment C was not quite different (P>0.05) with (2017c) stated that rumen environmental conditions have a
treatment D. The average of CFD in this study ranged from close relationship with the pH of the rumen fluid because
71.52-81.01%. Fermentation in tithonia can increase CFD. the high and low pH in the rumen will affect the rumen
It is because fermentation can stretch the bonds of fiber microbial activity.
fraction components to make rumen microbes work more
optimally. The analysis of variance showed that tithonia
fermentation with different types of microbes and
The highest CFD value in this study was found in incubation time gave no significant effect (P>0.05) on the
treatment B, which was 81.01%. The increase in CFD in production of rumen fluid VFA. It indicated that feed
treatment B was due to the crude fiber content of the fermentation using L. plantarum and A. ficuum did not
material in treatment B, which was lower than the other affect rumen microbial activity in producing VFA in the
treatments with 16.52%, so that rumen microbial activity in form of acetic acid, propionic acid, and butyric acid. The
digesting crude fiber was more optimal. According to Yanti production of VFA in this study ranged from 116.67 mM
et al. (2021) and Jamarun et al. (2018), feed ingredients 135 mM; this value was still in the range of VFA
with low crude fiber will generally be easier to digest concentrations under normal conditions. McDonald et al.
because microbes easily penetrate the cell walls of these (2010) stated that the normal range for total VFA was 70-
materials. On the contrary, the higher the crude fiber 150 mM.
content contained in a feed ingredient, the cell walls will be

318 Dasar Nutrisi Ruminansia (Edisi ke II)

PAZLA et al. Nutritional evaluation of Tithonia diversifolia using fermentation technology 3941

Table 6. Digestibility of dry matter, organic matter, crude protein, degradation of CP in the rumen will increase the
and crude fiber from fermented tithonia concentration of NH3 and vice versa

Nutritional digestibility (%) The lowest NH3 concentration was found in treatment
D, which was 9.21. The low concentration of NH3 in
Treatments Dry Organic Crude Crude treatment D was due to the low availability of N in the
feed; this was indicated by the low crude protein content of
matter matter Protein fiber the material, which was 27.56. Mayulu (2014) and
Wulandari et al. (2017) stated that the lower the crude
A 65.73 66.22 69.70 78.11b protein content of the ration, the production of NH3 would
also decrease. The NH3 production produced in each
B 66.86 67.36 71.50 81.01a treatment in this study ranged from 9.21 to 14.31. This
value is still in the normal range for rumen microbial
C 62.21 66.03 69.57 71.52c activity and growth. The opinion of Satter and Slyter
(1973) stated that the minimum concentration of ammonia
D 66.01 66.69 67.78 73.79c required for microbial protein synthesis is 5 mg/100ml
rumen fluid. The study concluded that fermented tithonia
SE 0.32 0.36 0.62 0.69 using A. ficuum with an incubation period of 7 days
improve the quality of tithonia. It is seen from the content
Note: Different superscripts in the same column show of CP (31.02%), CF (16.52%), phytase enzyme activity
(37.36 U/ml), DMD (66.86%), OMD (67.36%), CFD
significantly different effects (P<0.05) (81.01%), CPD (70.37%), VFA production 135 mM and
NH3 concentration 14.31 mg/100 ml, and pH value 6.72
Table 7. Rumen fluid characteristics from fermented tithonia which is very suitable for rumen microbial growth.

Treatments Rumen fluid characteristics
A
pH VFA (mM) NH3 (mg/100ml)

6.79 130 12.18ab
14.31a
B 6.72 135

C 6.85 116.67 10,48b

D 6.83 123.33 9.21b

SE 0.03 6.96 0.86

Note: Different superscripts in the same column show very

significant different effects (P<0.05)

The insignificant difference in VFA production was ACKNOWLEDGEMENTS
also due to the relatively similar composition, feeding
level, and physical form of the feed. Storm and Kristensen Thank you to the Indonesian Ministry of Research and
(2010) stated that the design of VFA in the rumen changes Technology/ National Research and Innovation Agency for
with differences in physical form, feed composition, level funding this research with contract number
and frequency of feeding, and processing. High or low 266/SP2H/LT/DPRM/2021.
VFA production can be used as a measure of the
fermentability of a feed ingredient. Jones and Murphy REFERENCES
(1989) stated that the higher the fermentability level of the
feed, the greater the VFA produced. Alami NH, Kuswytasari ND, Zulaika E, Shovitri M. 2017. Optimization
of cellulase production by candida G3. 2 from the rhizosphere of
The analysis of variance showed that tithonia Gunung Anyar mangrove, Surabaya. Proc Intl Conf Green Technol 8:
fermentation with different types of microbes and 399-406.
incubation time had a significantly different effect (P<0.05)
on the production of rumen fluid NH3. The results of the AOAC. 2010. Official Methods of Analysis. Association of Analytical
DMRT further test showed that treatment A was not Chemists, Arlington, Virginia, USA.
significantly different (P>0.05) from treatments B, C, and
D. However, Treatment B showed a significant difference Arief, Rusdimansyah, Sowmen S, Pazla R, Rizqan. 2020. Milk production
(P<0.05) with treatments C and D. Also, treatment C and quality of Etawa crossbred dairy goat that given Tithonia
showed the effect was not significantly different from diversifolia, corn waste and concentra tes based palm Kernel cake.
treatment D. Biodiversitas 21 (9): 4004-4009. DOI: 10.13057/biodiv/d210910

The highest NH3 production was found in treatment B, Arief, Simel S, Rusdimansyah, Pazla R. 2019. Ration digestibility based
namely, 14.31 mg/100ml. The high output of NH3 in on palm oil industry by products, tithonia (Tithonia diversifolia) and
treatment B indicated that the rumen microbial activity was corn waste for Etawa crossbred dairy goat. Pak J Nutr 18 (8): 733-
running well. NH3 concentration means the fermentability 738. DOI: 10.3923/pjn.2019.733.738
of feed related to protein digestibility, rumen microbial
activity, and microbial population (Pazla et al. 2018b). In Arief, Jamarun N, Satria B, Pazla R. 2021. Milk quality of Etawa dairy
addition, the high production of NH3 in treatment B was goat fed palm kernel cake, tithonia (Tithonia diversifolia), and Sweet
due to the increased degradation of crude protein in the potato leaves (Ipomoea batatas L). IOP Conf Ser Earth Environ Sci
rumen. NH3 concentration was strongly influenced by 709 (1): 012023. DOI: 10.1088/1755-1315/709/1/012023.
protein solubility. The more soluble the protein, the easier
it will be for rumen microbes to degrade it; the CPD Aye PA. 2016. Comparative nutritive value of Moringa oleifera, Tithonia
indicates this in treatment B, which was higher than in diversifolia, and Gmelina arborea leaf meals. Am J Food Nutr 6 (1):
other treatments (Table 2). It is following the opinion of 23-32.
Bannink et al. (2016), which states that. The higher the
Bannink A, Henk J, Van L, Jennifer L, Ellis, James F. 2016. The
contribution of mathematical modeling to understanding dynamic
aspects of rumen metabolism. Front Microbiol 7: 1-16. DOI:
10.3389/fmicb.2016.01820.

Bhatia KS, Yang HY. 2017. Reviews: Microbial production of volatile
f Environ Sci
Bio/Technol. DOI: 10.1007/s11157-017-9431-4.

Hafis A. 2019. Produksi titonia (Tithonia diversifolia) sebagai pakan
hijauan dengan jenis pupuk berbeda pada tanah ultisol. [Thesis].
Fakultas Peternakan Universitas Andalas, Padang. [Indonesian]

319Dasar Nutrisi Ruminansia (Edisi ke II)

3942 22 (9): 3936-3942, September 2021

Jamarun N, Agustin F, Pazla R, Oktiacahyani N. 2017b. Effects of Oluwasola TA, Dairo FAS. 2016. Proximate composition, amino acid

phosphor (P) supplementation in combination with calcium (Ca) and profile, and some anti-nutrients of Tithonia diversifolia cut at two
manganese (Mn) during oil palm frond fermentation by different times. Afr J Agric Res 11 (38): 3659-363. DOI:
Phanerochaete chrysosporium on fiber fractions content. In 10.5897/AJAR2016.10910.

Proceedings of The 5th International Seminar of Animal Nutrition Pazla R, Jamarun N, Zain M, Arief. 2018b. Microbial protein synthesis
and Feed Sciences, Mataram: Indonesian Association of Nutrition and in vitro fermentability of fermented oil palm fronds by
And Feed Scientist. [Indonesian] Phanerochaete chrysosporium in combination with tithonia (Tithonia
Jamarun N, Elihasridas, Pazla R, Fitriyani. 2017a. In vitro nutrients diversifolia) and elephant grass ( Pennisetum purpureum). Pak J Nutr
digestibility of the combination tithonia (Tithonia Diversifolia) and 17 (10): 462-470. DOI: 10.3923/pjn.2018.462.470.
Napier Grass (Pennisetum purpureum). In The 7th International
Pazla R, Zain M, Ryanto I, Dona A. 2018a. Supplementation of minerals
Seminar on Tropical Animal Production, Yogyakarta . [Indonesian]
Jamarun N, Pazla R, Yanti G. 2021. Effect of boiling on in-vitro nutrients (phosphorus and sulfur) and Saccharomyces cerevisiae in a sheep diet
based on a cocoa by-product. Pak J Nutr 17 (7): 329-335. DOI:
digestibility, rumen fluid characteristics, and tannin content of 10.3923/pjn.2018.329.335.

mangrove (Avicennia marina) leaves as animal feed. Intl Conf Green Putra AYT, Rosida, Azmir K. 2019. Chemical and sensory characteristics
Agro-Ind Bioecon. DOI: 10.1088/1755-1315/733/1/012106 of sorghum (Sorghum bicolor) tapai with traditional packaging. Food
Jamarun N, Pazla R, Zain M, and Arief. 2019. Comparison of in vitro ScienTech J 1 (2): 92-99. DOI: 10.33512/fsj.v1i2.7096.

digestibility and rumen fluid characteristics between the tithonia Satter LD, Slyter LL. 1973. Effect of ammonia concentration on rumen
(Tithonia diversifolia) with elephant grass (Pennisetum purpureum). microbial protein production in vitro. Br J Nutr 32: 199-207. DOI:
International Conference on Animal Production for Food 10.1079/BJN19740073.
Sustainability. IOP Conf Ser Earth En viron Sci 287: 1-5. DOI:
10.1088/1755-1315/287/1/012019. Saylor BA, Casale F. 2020. Effect of microbial inoculation and particle
Jamarun N, Zain. M. 2013. Dasar Nutrisi Ruminansia. Jasa Surya Press, size on fermentation profile, aerobic stability, and ruminal in situ
starch degradation of high-moisture corn ensiled for a short period. J
Padang. [Indonesian]
Jamarun N, Zain M, Arief, Pazla R. 2017c. Effects of calcium (Ca), Dairy Sci 103 (1): 1-16. DOI: 10.3168/jds.2019-16831.
Selle PH, Ravindran P, Caldwell RA, Bryden WL. 2021. Phytate and
phosphorus (P), and manganese (Mn) supplementation during oil
. Nutr Res Rev 13 (2):
palm frond fermentation by Phanerochaete chrysosporium on rumen
fluid characteristics and microbial protein synthesis. Pak J Nutr 16 255-278. DOI: 10.1079/095442200108729098
(6): 393-399. DOI: 10.3923/pjn.2017.393.399. Shieh TR, Ware JH. 1968. Survey of microorganisms for the production

Jamarun N, Zain M, Arief, Pazla R. 2018. Populations of rumen microbes of extracellular phytase. Appl Microbiol 16 (9): 1348-1351. DOI:
and the in vitro digestibility of fermented oil palm fronds in
combination with tithonia (Tithonia diversifolia) and elephant grass 10.1128/am.16.9.1348-1351.1968
Steel RGD, Torrie JH. 2002. Principle and Procedures of Statistics: A
(Pennisetum purpureum). Pak J Nutr 17 (1): 39-45. DOI:
10.3923/pjn.2018.39.45. Biometrical Approach. 3rd ed. McGraw Hill Book, New York.
Jones S, Murphy MR. 1989. Production of volatile fatty acids in the
rumen and cecum- Storm AC, Kristensen NB. 2010. Effects of particle size and dry matter
and forage physical form. J Dairy Sci 72 (2): 485-492. DOI: content of a total mixed ration on intraruminal equilibration and net
10.3168/jds.S0022-0302(89)79130-X. portal flux of volatile fatty acids in lactating dairy cows. J Dairy Sci
93 (9): 4223-4238. DOI: 10.3168/jds.2009-3002.
Maryanti T. 2015. Isolasi dan karakteristik kapang endofit dari ranting
tanaman parijoto (Medinilla speciosa Reinw. Ex Blume) dan uji Sumenger M, Dincer S, Kaya A. 2013. Production and characterization of
aktifitasnya sebagai antibakteri. [Thesis]. UIN Syarif Hidayatullah , phytase from Lactobacillus plantarum. Food Biotechnol 27 (2): 105-

Jakarta. [Indonesian] 118. DOI: 10.1080/08905436.2013.781507.
Maulana Z. 2019. Pengaruh dosis inoculum Aspergillus ficuum dan lama Susana IWR, Tangenjaya B, Hastiono S. 2000. Seleksi kapang penghasil

fermentasi terhadap aktifitas selulase, serat kasar dan daya cerna serat enzim fitase. Jurnal Ilmu Ternak dan Veteriner 5 (1): 1-6.

kasar ampas susu kedelai fermentasi. [Thesis]. Universitas Andalas, [Indonesian]
Padang. [Indonesian] Susanti D, Jamarun N, Agustin A, Astuti A, Yanti G. 2020. Kecernaan in-
Mayulu H. 2014. The nutrient digestibility of local sheep fed with amofer
palm oil byproduct-based complete feed. Int J Sci Eng 7: 106-111. vitro fraksi serat kombinasi pu cuk tebu dan titonia fermentasi sebagai
DOI: 10.12777/ijse.7.2.106-112.
McDonald P, Edwards R, Greenhalg JFD, Morgan CA. 2010. Animal pakan ruminansia. Agripett 20: 86-95. DOI:
10.17969/agripet.v20i1.16040. [Indonesian]
nutrition. Pearson Education Limited, England. Tilley JMA, Terry RA. 1963. A two-stage technique for the in vitro
Mingardon F, Bagert JD, Maisonnier C, Devin L. Trudeau, Frances H. A. digestion of forage crops. J Brit Grassland Soc 18: 104-111. DOI:
10.1111/j.1365-2494.1963.tb00335.x.
2011. Comparison of family 9 cellulases from mesophilic and Van Soest PJ, Robertson JB, Lewis BA. 1991. Methods for dietary fiber,

thermophilic bacteria. Appl Environ Microbiol 77 (4): 1436-1442. neutral detergent fiber, and nonstarch polysaccharides in relation to
DOI: 10.1128/AEM.01802-10. animal nutrition. J Dairy Sci 74 (10): 3583-3597. DOI:
Mirnawati, Djulardi A, Marlida Y. 2013. Improving the quality of palm 10.3168/jds.S0022-0302(91)78551-2.

kernel cake through fermentation by Eupenicillium javanicum as Wulandari, Budi PW, Cuk TN, Ali A. 2017. In vitro degradation and
poultry ration. Pak J Nutr 12 (12): 1085-1088. DOI: rumen fermentation characteristics of soybean meal protected with
10.3923/pjn.2013.1085.1088. different levels of formaldehyde. in The 7th International Seminar on

Mirnawati, Rizal Y, Marlida Y, Kompiang PI. 2010. The role of humic Tropical Animal Production, Yogyakarta. [Indonesian]
acid in palm kernel cake fermented by Aspergillus niger for poultry Yanti G, Jamarun N, Elihasridas, Astuti T. 2021. Quality improvement of
ration. Pak J Nutr 9 (2): 182-185. DOI: 10.3923/pjn.2010.182.185.
sugarcane top as an animal feed with biodelignification by

Phanerochaete chrysosporium fungi on in-vitro digestibility. In
ICOMSET2020. DOI: 10.1088/1742-6596/1940/1/012063.

320 Dasar Nutrisi Ruminansia (Edisi ke II)

1

321Dasar Nutrisi Ruminansia (Edisi ke II)

2

322 Dasar Nutrisi Ruminansia (Edisi ke II)

Table 1. Average laccase enzyme activity (U/mL).

Type of fungi Fermentation time B3
B1 B2 2.68±0.83a
1.32±0.08b
A1 1.16±0.55b 1.62±0.13b

A2 1.00±0.01b 1.18±0.05b

SE 0.25

Table 2. Crude protein content sugarcane top biodelignification (%).

Type of fungi Fermentation time B3 Average
B1 B2
8.33
A1 8.56 9.23 8.69 8.75

A2 8.45 8.57 9.22

Average 8.51 8.90 8.95

3

323Dasar Nutrisi Ruminansia (Edisi ke II)

Table 3. Average lignin content degradation (%).

Type of fungi Fermentation time Rataan

A1 B1 B2 B3 19.01±4.91a
A2 9.72±2.18b
Rataan 13.36 18.35 25.32
14.37
6.67 11.61 10.88

10.02±3.34b 14.98±3.37ab 18.10±7.2a

Nyakira B, Tuitoek J, Onjoro,P, Ambula 2010 Evaluation of the Nutritive Value of Sugar Cane
Tops, Mulberry Leaves (M. Alba) and Calliandra (C. Calothyrsus) as Feed Supplements for
Goats. J. of Animal Science Advances. 5. 10.5455/jasa.20150315015511

Gutiérrez B, Ortiz-Muñiz B, Rodriguez G, Javier, Cardenas, Angel, González, José, Aguilar-
Uscanga, Maria 2015 Bioethanol production from hydrolyzed sugarcane bagasse
. Renewable Energy. 74. 399 405.
10.1016/j.renene.2014.08.030

Sharma S, Duggirala S 2020 Production of Lignolytic and Cellulolytic Enzymes by using
Basidiomycetes Fungi in the Solid State Fermentation of Different Agro-Residues. 15. 17

Kenealy W, Jeffries, Thomas 2003 Enzyme Processes for Pulp and Paper: A Review of Recent
Developments. ACS Symposium Series. 845 : 210-239. 10.1021/bk-2003-0845.ch012

Aleksieva P. Tchorbanov, Bozhidar, Nacheva, Lilyana 2010 High-Yield Production of Alpha-
Galactosidase Excreted from Penicillium Chrysogenum and Aspergillus Niger. Biotechnology
& Biotechnological Equipment - biotechnol biotechnol equip. 24. 1620-1623.
10.2478/V10133-010-0015-5

Bentley R, Bennett JW 2008 A ferment of fermentations: Reflections on the production of
commodity chemicals using microorganisms. Journal Applied Microbiology 63: 1-32

Mojsov K 2010 Experimental investigations of submerged fermentation and synthesis of
pectinolytic enzymes by Aspergillus Niger : Effect of inoculum size and age of spores.
App.Technologies and Innovations. 2. 40-46. 10.15208/ati.2010.13

Zhang, Yi-Heng P.Hong, Jiong, Ye, Xinhao 2009 Cellulase Assays. Methods in molecular biology
(Clifton, N.J.). 58. 13-31. 10.1007/978-1-60761-214-8_14

Thakkar, Asha, Pandya DC, Bhatt, Shreyas 2020 Optimization of Laccase Enzyme Production
by Amesia atrobrunnea A2: A First Report. Biosciences, Biotechnology Research Asia. 17.
65-72. 10.13005/bbra/2810

Senthivelan T, Kanagaraj J, Rames C, Panda T, Narayani 2019 Screening and production of a

4

324 Dasar Nutrisi Ruminansia (Edisi ke II)

potential extracellular fungal laccase from Penicillium chrysogenum: Media optimization by
response surface methodology (RSM) and central composite rotatable design (CCRD),
Biotechnology Reports. Volume 23. ISSN 2215-017X,
https://doi.org/10.1016/j.btre.2019.e00344
Sharma K, Kuhad R 2008 Laccase: Enzyme revisited and function redefined. Indian journal of
microbiology. 48. 309-16. 10.1007/s12088-008-0028-z
Mallak AM, Lakzian A, Khodaverdi E, Haghnia G, Mahmoudi S 2020 Effect of Pleurotus
ostreatus and Trametes versicolor on triclosan biodegradation and activity of laccase and
manganese peroxidase enzymes. Microbial Pathogenesis. Vol 149. 104473. ISSN 0882-4010.
https://doi.org/10.1016/j.micpath.2020.104473
Oboh C, Elusiyan CA 2007 Changes in the nutrient and anti nutrient content of micro-fungi
fermented cassava flour produced from low and medium-cyanida variety of cassava tubbers.
J. African Journal of Biotechnology 6:2150-2157
Schoenherr, Stefan, Ebrahimi, Mehrdad, Czermak, Peter 2018Lignin Degradation Processes and
the Purification of Valuable Products. 10.5772/intechopen.71210
Kumar A, Chandra R, 2020 Ligninolytic enzymes and its mechanisms for degradation of
lignocellulosic waste in environment, Heliyon, Vol 6, Issue 2, ISSN 2405-8440.
https://doi.org/10.1016/j.heliyon.2020.e03170
Reid I. 2011 Biodegradation of lignin. Canadian Journal of Botany. 73. 1011-1018. 10.1139/b95-
351

5

325Dasar Nutrisi Ruminansia (Edisi ke II)

Reproduced with permission of copyright owner. Further reproduction
prohibited without permission.

326 Dasar Nutrisi Ruminansia (Edisi ke II)

Volume 22, Number 11, November 2021 ISSN: 1412-033X
Pages: 4794-4798 E-ISSN: 2085-4722
DOI: 10.13057/biodiv/d221111

Degradation of phytic acid from tithonia (Tithonia diversifolia) leaves
using Lactobacillus bulgaricus at different fermentation times

RONI PAZLA , GUSRI YANTI1, NOVIRMAN JAMARUN1, ARIEF2, ELIHASRIDAS1,
LARAS SUKMA SUCITRA1

1Department of Nutrition Science and Feed Technology, Faculty of Animal Husbandry, Universitas Andalas. Jl. Kampus, Limau Manis, Padang 25163,
West Sumatra, Indonesia. Tel./fax.: +62-751-71464, email: [email protected]

2Department of Technology and Animal Production, Faculty of Animal Husbandry, Universitas Andalas. Jl. Kampus, Limau Manis, Padang 25163, West
Sumatra, Indonesia

Manuscript received: 31 August 2021. Revision accepted: 14 October 2021.

Abstract. Pazla R, Yanti G, Jamarun N, Arief, Elihasridas, Sucitra LS. 2021. Degradation of phytic acid from tithonia (Tithonia
diversifolia) leaves using Lactobacillus bulgaricus at different fermentation times. Biodiversitas 22: 4794-4798. The aim of this study is
to reduce high level of phytic acid in tithonia (Tithonia diversifolia) leaves by fermentation technique using Lactobacillus bulgaricus. A
completely randomized design consisting of four treatments (fermentation time) i.e. T2: 2 days, T3: 3 days, T4: 4 days and T5: 5 days
and experiment was performed in four replicates. The parameters measured were pH, phytase enzyme activity, total bacterial colony,
phytic acid content, and phytic acid degradation. The results showed that the fermentation time had a significant effect (P<0.05) on pH,
phytase enzyme activity, total bacterial colony, phytic acid content, and phytic acid degradation. The conclusion of this study is that on
fifth day (Treatment T5) fermented tithonia leaves produced the lowest levels of phytic acid (3.48 mg/ 100g) with the highest level of
degradation (64.81%).

Keywords: Fermentation times, Lactobacillus bulgaricus, phytic acid, ruminant feed, Tithonia diversifolia.

INTRODUCTION chance of feed phytic acid escaping to post-rumen. This
condition negatively affects because phytic acid as a
Tithonia diversifolia (Hamsley) A.Gray is a forage chelating agent can bind some minerals or compounds
protein source for ruminants has been studied by essential to ruminants. Rumen microbes produce many
researchers. Pazla (2018) reported that the leaves and stem phytase enzymes, but very few enzymes are present in the
of tithonia plant contain 84.01% organic matter, 22.98% post-rumen. It is difficult to break down phytic acid into
crude protein, 18.17% crude fiber and 34.59% cellulose. free inositol and phosphate (Park et al. 1999). Non-
Tithonia leaves are rich in essential amino acids, such as ruminant livestock, such as pigs and chickens do not have
methionine, leucine, isoleucine, and valine which are phytase enzymes, so they cannot degrade phytic acid to
important for the growth of rumen microbes (Oluwasola digestible phosphorus (Greiner and Konietzny 2011).
and Dairo 2016). Tithonia leaves contain macro minerals, Phytic acid content in pig feed is usually between 7 and 10
such as Ca and Mg and several micro-minerals that are g/kg of diet (Selle et al. 2009).
very beneficial for rumen microbial activity and growth
(Mahecha and Rosales 2005). The phosphorus in phytic acid is difficult to digest.
This causes phosphorus cannot be utilized by rumen
Besides containing good nutritional potential, tithonia microbes and livestock bodies. The phosphorus will be
also contains many anti-nutritional substances, which are wasted through feces and pollute the environment. The
harmful to livestock health (Aye 2016; Jamarun et al. body can use elemental P in the degradation of phytic acid.
2020). Phytic acid is the most anti-nutritional substance in Degradation is the process of breaking the bonds of myo-
tithonia (Fasuyi et al. 2010). Phytic acid (C6H18O24P6 or inositol group with the phosphate group. The released
IP6) is a phosphate-bound myo-inositol ring (Turk 1990; phosphate is a source of phosphorus for the body and
Susanti 2012). The high level of phytic acid in tithonia rumen microbes (Pazla et al. 2018, 2020).
produces a bitter taste, which diminishes the flavor of
plant, causing the animal to consume less of it (Pazla et al. Fermentation technology can be used to degrade phytic
2021a). Phytic acid has also been reported to bind proteins, acid. Lactobacillus bulgaricus which produces phytase
carbohydrates and minerals essential for rumen microbes enzymes used as a phytate-degrading microorganism
such as P, Mg and Zn (Lai et al. 2013; Selle et al. 2021). (Mohamed et al. 2011; Sripo et al. 2016). The aim of this
Feeds containing high phytic acid have a fast rate of study is to reduce the high level of phytic acid content in
degradation (< 8 hours) in the rumen, leading to limited tithonia (T. diversifolia) leaves using L. bulgaricus at
phytic acid degradation by rumen microbes and a greater different fermentation times.

327Dasar Nutrisi Ruminansia (Edisi ke II)

PAZLA et al. Degradation of phytic acid from tithonia leaves using Lactobacillus bulgaricus 4795

MATERIALS AND METHODS Measurement of phytic acid content
One gram of sample was homogenized with 50 mL of
Experimental site
Lactobacillus bulgaricus culture was collected from the 0.5 M HNO3 using a shaker for 2 hours, then strained. The
filtrate was used for the measurement of phytic acid. 0.9
Livestock Technology Laboratory, Faculty of Animal mL of 0.5 N HNO3, and 1 mL of FeCl3 (containing iron
Husbandry, Bogor Agricultural University, Indonesia. ions 50 micrograms/mL) were added to 0.5 mL of filtrate,
Tithonia leaves were taken from Padang Panjang, West covered with aluminum foil, and then kept in boiling water
Sumatra, Indonesia. The fermentation process was carried for 20 minutes. Cool the tubes with water, adding 5 mL of
out in the Technology and Feed Industry Laboratory, amyl alcohol and 1 mL ammonium thiocyanate (10%).
Faculty of Animal Husbandry, Andalas University, Tubes were centrifuged for 10 minutes at 10,000 rpm and
Indonesia. absorbance was measured at 465 nm using spectrophotometry.

Procedures Measurement of phytase enzyme activity
Rejuvenation of Lactobacillus bulgaricus A 0.15 mL of enzyme was taken in a test tube, then 0.6

Lactobacillus bulgaricus was grown on Potato Dextrose mL of 0.1 M Tris HCl buffer containing Ca-phytate, CaCl
Agar (PDA) media. 4.04 g of PDA media was dissolved 2, 0.75 TCA 5% was added and incubated at 37°C for 30
with 175 mL of distilled water in an Erlenmeyer flask, and min. After that, 1.5 mL molybdate color reagent was added
then heated on a hot plate until boiling. The flask was and homogenized by vortex. Absorbance of the
covered with cotton plug and sealed with aluminum foil, characteristic color was measured at 700 nm against blank
then sterilized in autoclave for 30 minutes at 121°C. After using spectrophotometer.
cooling, media was inoculated with 5 mL of L. bulgaricus
and incubated in a shaker incubator at 37°C for 48 hours. Data analysis
Data were statistically analyzed by analysis of variance
Tithonia fermentation process
A 50 g of tithonia flour was mixed with 80 mL of with four treatments (fermentation time) and four
repetitions using SPSS software version 20.0. Differences
distilled water and put in a plastic bag, then homogenized. between treatments means were determined by Duncan's
Then plastic bags were sterilized in an autoclave at 121°C Multiple Range Test. The level of statistical significance
for 30 minutes and cool down. After that, 1.5 mL of L. was set at p < 0.05.
bulgaricus was inoculated into broodstock and
homogenized. All samples were tightly covered with tape RESULTS AND DISCUSSION
and plastic wrap and stored at room temperature for further
investigation. Fermentation was done for four days i.e. T2: Total bacterial colonies
2 days, T3: 3 days, T4: 4 days and T5: 5 days. The statistical analysis results showed that the treatment

Total bacterial colonies had a significantly different effect (P<0.05) on the total
Weighed 10 g of the fermented sample, dissolved in 90 bacterial colonies. Based on Table 1, the average entire
bacteria colony ranged from 8.50-31×109 U/mL. L.
mL of distilled water (101). Prepared 8 test tubes bulgaricus has four growth phases: the lag phase,
(containing 9 mL of distilled water). Pipette 1 mL of exponential phase, stationary phase, and death phase
broodstock liquor (101), transferred to test tube 102 and (Malaka 1997). L. bulgaricus reached its optimum growth
then homogenized, continued until 109 dilution. Next, on second day of fermentation with a total bacterial colony
pipette 1 mL of solution from test tube 8 (109) transferred of 31×109 U/mL. Bacterial growth was decreased by the
into a petri dish. Add 10 mL of MRS Agar (De man 5th day of fermentation with whole colony of 8.5×109
Rogosa and Sharpe) medium, homogenize. Incubated at U/mL. The lag phase of L. bulgaricus occurred on 24 hours
37oC for 24 hours. Count the total bacterial colonies. of incubation. After that, the bacteria entered an
exponential phase and with the maximum number of
Making phytase media colonies appeared on second day. There was a sharp
The purpose of making phytase media was to measure decrease in total bacterial colonies on the third day. This
may be due to decreased nutrient levels and accumulation
the activity of phytase enzyme. For this 1.5 g of glucose, of toxic products that interfere with bacterial cell division.
1.5 g of MRS Agar, 0.05 g of MgSO4, 0.02 g of MnSO4, Bacterial death rate was more significant on the 5th day of
0.05 g of KCl, 0.5 g of Ca-Phytate, 0.5 g of NH4NO3 and incubation, leading to a reduction in bacterial population.
0.001 g of FeSO4 was put in a flask and added 100 mL of The total bacterial colony in this study was higher than the
distilled water. Then boil it on a hot plate, cover with results of Malaka and Laga (2005) research, which was 4.9
aluminum foil and sterilized at 121°C for 30 minutes in × 109, and lower than the Sripo et al. (2016) study, which
autoclave. Media was poured into a sterilized petri dish and was 7x1010 U/mL. The difference is caused by the type of
inoculated by spread with 1 mL of L. bulgaricus substrate and the length of fermentation. The total colonies
broodstock. The plates were then tightly covered with of L. bulgaricus bacteria at different fermentation times can
plastic wrap and stored at room temperature for 24 hours. be seen in Figure 1.

328 Dasar Nutrisi Ruminansia (Edisi ke II)

4796 22 (11): 4794-4798, November 2021

Phytase enzyme activity activity by the fifth day of fermentation. Phytase
Table 1 showed that the length of fermentation had a (myoinositol-hexakisphosphate-3-phosphohydrolase) is an
enzyme that catalyzes myo-inositol hexakisphosphate
significant effect (P<0.05) on phytase enzyme activity. The (phytate) to inorganic orthophosphate and a lower series of
highest i.e. 15.02 U/mL phytase enzyme activity occurred phosphoric (inositol Penta phosphate to monophosphate)
on second day of fermentation (T2 treatment) and and finally to free Myo-inositol (Selle and Ravindran
2007). Phytase can release phosphorus bound to phytate to
decreased with the length of fermentation time. The lowest be available and can be utilized by livestock (Hidayat
i.e. 5.06 U/mL phytase enzyme activity was recorded on 2016). The duration of fermentation results in the
fifth day (T5). The optimal movement of phytase enzyme accumulation of phytate degradation by L. bulgaricus
on second day of fermentation (T2) was due to the high bacteria which leads to a decrease in phytate content on the
fifth day of fermentation. In this study, phytic acid
nutrient content of the substrate for the growth of L. degradation was higher than Sripo et al. (2016) research,
bulgaricus. The total number of bacterial colonies reached 54.8% in fermented black glutinous rice and lower than the
an optimum growth of 31 × 109 U/mL on the second day of results of Mohamed et al. (2011) study, which was 69.2%
fermentation. Bacterial populations can result in high in fermented mung bean using L. bulgaricus. These
enzyme activity. Enzyme activity is strongly influenced by differences in results may be due to the different types of
substrates. The phytic acid content of fermented tithonia
the nutrient content of the substrate (Poernomo et al. 2017; leaves by L. bulgaricus bacteria at different fermentation
Pazla et al. 2020). times can be seen in Figure 3.

The longer the fermentation time, the lower the nutrient Figure 1. Total bacterial colonies of fermented tithonia leave by
content of the substrate as bacteria use up all the food for Lactobacillus bulgaricus at different fermentation times. * T2 (2
days), T3 (3 days), T4 (4 days) and T5 (5days)
their activity and growth. Putra et al. (2019) stated that the
fermentation process carried out by microorganisms can
remodel carbohydrates into alcohol, organic acids, H2O,
and CO2. Also, Mirnawati et al. (2013) added that the

length of fermentation time depletes the nutrients in the
substrate as the microorganisms utilized it for their growth.
Phytase enzymes produced during the fermentation process
contribute significantly to the level of phytic acid

substrates. The higher the activity of phytase enzyme
produced by microbes, the lower the phytic acid content
(Reddy and Pierson 1994). In this study, phytase enzyme
activity from L. bulgaricus was higher, but the activity of

phytase enzyme produced by Lactobacillus plantarum in
the fermented tithonia plant was 6.68-11.71 U/mL (Pazla et
al. 2021b). The difference in values may be due to the type
of lactic acid bacteria used. The phytase enzyme activity
of L. bulgaricus bacteria at different fermentation times can

be seen in Figure 2.

Phytic acid content and phytic acid degradation Figure 2. Phytase enzyme activity of fermented tithonia leaves by
Statistical analysis showed that the treatment duration Lactobacillus bulgaricus at different fermentation times. * T2 (2
days), T3 (3 days), T4 (4 days) and T5 (5days)
of fermentation had a significantly different effect (P<0.05)

on phytic acid content and phytic acid degradation (Table 1
and Table 2). The T2 treatment (2 days of fermentation)
showed highest phytic acid content of 4.85 mg/100g with a
degradation rate of 51.03%. The lowest (3.48 mg/100g)

phytic acid content was observed on fifth day of
fermentation (T5), with a degradation rate of 64.81%.
Phytic acid content decreased with increasing fermentation
time. This is due to the phytase enzyme produced by L.
bulgaricus bacteria during the fermentation process. Table

1 showed that L. bulgaricus carried phytase enzyme

Table 1. Total bacterial colonies, phytase enzyme activity, phytic acid content and pH of fermented tithonia leaves by Lactobacillus
bulgaricus at different fermentation times

Parameters Treatments T5 SEM

*T2 T3 T4
Total bacterial colonies (109) 31d ±5.48 16.75c±2.22 12.75b±2.22 8.5a±1.29
0.81

Phytase enzyme activity (U/mL) 15.02d±0.26 7.32c ±0.35 5.94b ±0.18 5.06a ±0.25 0.07

phytic acid content (mg/100 g) 4.85d±0.17 3.99bc±0.18 3.89 b±0.13 3.48a±0.34 0.05

pH 6.97c±0.06 6.98c ±0.04 7.09a±0.02 7.29ab ±0.09 0.02

Note: Different superscripts (a,b,c,d) in the same row are significantly different (P<0.05). * T2: 2 days, T3: 3 days, T4: 4 days and T5: 5days

329Dasar Nutrisi Ruminansia (Edisi ke II)

PAZLA et al. Degradation of phytic acid from tithonia leaves using Lactobacillus bulgaricus 4797

pH fermentation Statistical analysis showed that fermentation time had a
pH is an acid-base condition of the fermentation significant effect (P<0.05) on pH. The lowest pH value was
found in T2 treatment, i.e. two days of fermentation. The
medium associated with the growth activity of pH value continued to increase till the fifth day of
microorganisms. The rate of microbial cell death can be fermentation i.e. 7.29. An increase in pH from 2nd day to
triggered when the pH is too low (acidic) or too high the 5th day of fermentation indicates that the fermentation
process is well underway. The pH can encourage the
(alkaline). The high mortality rate of microorganisms optimum activity of bacterial microorganisms. The degree
affects the speed of fermentation. The range of pH values of acidity at the beginning of fermentation process converts
obtained was 6.97-7.29. This pH value supported the organic matter into organic acids, besides the reshuffle of
growth of L. bulgaricus. The optimum pH for the phytic acid by the phytase enzyme that occurs during the
fermentation process, phytic acid is hydrolyzed into
development of L. bulgaricus is 5.5-6.2, and the growth inositol, thereby increasing the pH (Mittal et al. 2011). In
rate decreases in the early alkaline media (Sneath et al. addition, Sneath et al. (1986) reported that the L.
1986). Malaka and Laga (2005) reported that L. bulgaricus bulgaricus also reduced nitrate, causing an increase in the
could grow at alkaline pH (8.1). pH value of fermentation towards alkaline. Also, Hati
(2018) states that the increase in pH in the fermentation
Table 2. Phytic acid degradation of fermented tithonia leaves by process towards neutral occurs due to microorganisms that
Lactobacillus bulgaricus at different fermentation times convert organic acids. The pH of fermented tithonia by L.
bulgaricus at different fermentation times can be seen in
Phytic acid content (mg/100 g) Phytic acid Figure 4. From the results above, it can be concluded that
tithonia leaves fermented with L. bulgaricus on fifth day
Treatments Before After degradation showed the lowest phytic acid content 3.48 mg/100 g with
phytic acid degradation of 64.81%.
fermentation fermentation (%)
ACKNOWLEDGEMENTS
*T2 9.93±0.58 4.85±0.17 51.03a±4.26

T3 9.93±0.58 3.99±0.18 59.74b±3.10

T4 9.93±0.58 3.89±0.13 60.83bc±1.05

T5 9.93±0.58 3.48±0.34 64.81c±4.58

SEM 0.88

Note: Different superscripts (a, b, c, d) in the same column are

significantly different (P<0.05). * T2: 2 days, T3: 3 days, T4: 4

days and T5: 5days

The authors would like to thank the research and
community service institute Andalas University, Indonesia
who has funded this research with the contract number:
T/19/UN.16.17/PT.01.03/Pangan-RPB/2021 and also the
technicians of the technology and feed industry laboratory
technicians, Faculty of Animal Husbandry, Andalas
University and laboratory technicians from livestock
products technology, Faculty of Animal Husbandry, Bogor
Agricultural University, Indonesia.

Figure 3. The phytic acid content of fermented tithonia leaves by REFERENCES
L. bulgaricus at different fermentation times. * T2 (2 days), T3 (3
days), T4 (4 days) and T5 (5days) Aye PA. 2016. Comparative nutritive value of Moringa oleifera, Tithonia
diversifolia and Gmelina arborea leaf meals. Amer J Food Nutr 6 (1):
Figure 4. pH of fermented tithonia leaves by Lactobacillus 23-32.
bulgaricus at different fermentation times. * T2 (2 days), T3 (3
days), T4 (4 days) and T5 (5days) Fasuyi AO, Dairo FAS, Ibitayo FJ. 2010. Ensiling wild sunflower
(Tithonia diversifolia) leaves with sugar cane molasses . Lives Res
Rural Dev 22: 42. http://www.lrrd.org/lrrd22/3/fasu22042.htm [18-
10-2020].

Greiner R, Konietzny U. 2011. Phytase: biochemistry, enzymology and
characteristics relevant to animal feed use. In: Bedford MR, Partridge
GG (eds.). Enzymes in Farm Animal Nurition 2nd ed. AB Vista Feed
Ingredients Ltd., UK. DOI: 10.1079/9781845936747.0096.

Hati S. 2018. Pembuatan Pupuk Kompos Cair dari Limbah Rumah
Tangga sebagai Penunjang Mata Kuliah Ekologi dan Masalah
Lingkungan. [Thesis]. Islam Negeri Ar-Raniry Darussalam
University, Banda Aceh. [Indonesian]

Hidayat C. 2016. Utilisation of phytase to overcome phytic acid in broiler
diet. Wartazoa 26 (2): 57-68. DOI: 10.14334/wartazoa.v26i2.1326.

Jamarun N, Pazla R, Arief, Jayanegara A, Yanti G. 2020. Chemical
composition and rumen fermentation profile of mangrove leaves
(Avicennia marina) from West Sumatra, Indonesia. Biodiversitas 21
(11): 5230-5236. DOI: 10.13057/biodiv/d211126.

Lai LR, Hsieh SC, Huang HY, Chou C. 2013. Effect of lactic fermentation
on the total phenolic, saponin and phytic acid contents as well as anti -

330 Dasar Nutrisi Ruminansia (Edisi ke II)

4798 22 (11): 4794-4798, November 2021

colon cancer cell proliferation activity of soymilk . J Biosci Bioeng energy ratios. Adv Anim Vet Sci 9 (10): 1608-1615. DOI:

115 (5): 552-556. DOI: 10.1016/j.jbiosc.2012.11.022. 10.5713/ajas.17.0759.
Mahecha L, Rosales M. 2005. Nutritional value of the foliage of wild
Pazla R, Jamarun N, Zain M, Yanti G, Chandra RH. 2021b. Quality
sunflower (Botón de Oro; (Tithonia diversifolia [Hemsl] A.Gray) for
evaluation of tithonia (Tithonia diversifolia) with fermentation using
tropical animal production. Lives Res Rural Dev 17 (9): 100.
Malaka R. 1997. Effect of curdlan, a bacteria polysaccharide on the Lactobacillus plantarum and Aspergillus ficuum at different

physical properties and microstructure of acid milk curd by lactic acid incubation times. Biodiversitas 22 (9): 3936-3942. DOI:
fermenation. [Thesis]. Miyazaki University, Japan.
Malaka R, Laga R. 2005. Isolasi dan identifikasi Lactobacillus bulgaricus 10.13057/biodiv/d220940.
strain ropy dari yoghurt komersial. Sains dan Teknologi 5 (1): 50-58.
Pazla R, Zain M, Ryanto I, Dona A. 2018. Supplementation of minerals
[Indonesian]
Mirnawati, Djulardi A, Marlida Y. 2013. Improving the quality of palm (phosphorus and sulfur) and Saccharomyces cerevisiae in a sheep diet

ketnel cake through fermentation by Eupenicillium javanicum as based on a cocoa by-product. Pak J Nutr 17 (7): 329-335. DOI:

poultry ration. Pak J Nutr 12 (12): 1085-1088. DOI: 10.3923/pjn.2018.329.335.
10.14334/wartazoa.v28i3.1820.
Mittal A, Singh G, Goyal F, Yadaf A, Aneja KR, Gautam SK, Agarwal Poernomo AT, Isnaeni, Sugianto, Purwanto DA, Dewi AC, Suryagama D.

NK. 2011. Isolation and biochemical characterisation of acido- 2017. Pengaruh nutrisi pada produksi dan karakterisasi protease dari
thermophilic extracellular phytase producing bacterial strain for
potential application in pou ltry feed. J Microbiol 4: 273-282. bakteri termofilik isolat LS-1 lumpur sidoarjo. Jurnal farmasi dan
Mohamed R, Esmat A, Abou A, Gibriel AY, Rasmy N, Ferial M, Salem
A. 2011. Effect of legume processing treatments individually or in Ilmu Kefarmasian Indonesia 4 (2): 52-59. DOI:
combination on their phytic acid content. Afr J Food Sci Technol 2
10.20473/jfiki.v4i22017.51-58. [Indonesian]
(2): 36-46.
Oluwasola T A, Dairo FAS. 2016. Proximate composition, amino acid Putra AYT, Rosida, Khoirul A. 2019. Chemical and sensory characteristic

profile and some anti-nutrients of Tithonia diversifolia cut at two of sorghum (Sorghum bicolor) tapai with traditional packaging. Food

different times. Afr J Agric Res 11 (38): 3659-63. DOI: Sci Technol J 1 (2): 92-99. DOI: 10.33512/fsj.v1i2.7096.
10.5897/AJAR2016.10910.
Park WY, Matsui T, Konishi C, Kim SW, Yano F. 1999. Formaldehyde Reddy NR, Pierson MD. 1994. Reduction in Anti-nutritional and toxic

treatment suppresses ruminal degradation of p hytate in soyabean meal components in plant foods (A) by fermentation. Food Res Intl 27 (3):
and rapeseed meal. Br J Nutr 81 (6): 467-471. DOI:
10.1017/S0007114599000823. 281-290. DOI: 10.1186/s43014-020-0020-5.

Pazla R. 2018. Pemanfaatan Pelepah Sawit dan Tithonia (Tithonia Selle PH, Ravindran V, Caldwell RA, Bryden WL. 2021. Phytate and
diversifolia) dalam Ransum Kambing Peranakan Etawa untuk
Menunjang Program Swasembada Susu 2020. [Dissertation]. Andalas phytase: consequences for protein utilisation. Nutr Res Rev 13 (2):
University, Padang . [Indonesian]
255-278. DOI: DOI: 10.1079/095442200108729098.
Pazla R, Jamarun N, Agustin F, Zain M, Arief, Oktiacahyani N. 2020.
Effects of supplementation with phosphorus, calcium and manganese Selle PH, Ravindran V. 2007. Microbial phytase in poultry nutrition.

during oil palm frond fermentation by Phanerochaete chrysosporium Anim Feed Sci Technol 135 (1): 41. DOI:
on ligninase enzyme activity. Biodiversitas 21 (5): 1833-1838. DOI:
10.13057/biodiv/d210509. 10.1016/j.anifeedsci.2006.06.010.

Pazla R, Adrizal, Sriagtula R. 2021a. Intake, nutrient digestibility and Selle PH, Cowieson AJ, Ravindran V. 2009. Consequences of calcium
production performance of Pesisir cattle fed Tithonia diversifolia and
Calliandra calothyrsus-based rations with different protein and interactions with phytate and phytase for poultry and pigs. Livest Sci

124 (1-3): 126-141. DOI: 10.1016/j.livsci.2009.01.006.

Sneath PHA, Mair NS, Sharpe ME, Holt JG. 1986. Be of

Systematic Bacteriology. 2nd ed. Williams and Wilkins , Baltimore.

Sripo K, Phianmongkhol A, Wirjantoro TI. 2016. Effect of inoculum

levels and final ph values on the antioxidant properties of black

glutinous rice solution fermented by Lactobacillus bulgaricus. Intl

Food Res J 23 (5): 2207-2213.

Susanti ER. 2012. Degradasi Asam Fitat pada Kambing Peranakan

Etawah Laktasi yang Mendapat Ransum Bersuplemen Kedelai

Sangrai, Vitamin dan Mineral. [Thesis]. Bogor Agricultural Institute,

Bogor. [Indonesian]

Turk M. 1990. Cereal and Microbial Phytase, Phytase Degradation,

Mineral Binding and Absorption. [Dissertation]. Chalmers University

of Technology, Swedia.

331Dasar Nutrisi Ruminansia (Edisi ke II)

IOP Conference Series: Earth and Environmental Science

PAPER • OPEN ACCESS
To cite this article: N Jamarun et al 2021 IOP Conf. Ser.: Earth Environ. Sci. 733 012106
View the article online for updates and enhancements.

This content was downloaded from IP address 38.145.111.54 on 05/05/2021 at 02:07

332 Dasar Nutrisi Ruminansia (Edisi ke II)

Effect of boiling on in-vitro nutrients digestibility, rumen fluid
characteristics, and tannin content of mangrove (Avicennia
marina) leaves as animal feed

1

333Dasar Nutrisi Ruminansia (Edisi ke II)

In-vitro method: In vitro analysis was carried out to determine the digestibility of mangrove
leaves (Avicennia marina) for each feed treatment. The rumen process was stopped by immersing the
Erlenmeyer tube in ice water to stop the microbial activity, then measuring the pH using a pH meter.
The next step was to separate the supernatant from the residue. The mixture obtained from in vitro
analysis was centrifuged for 30 minutes at a speed of 3000 rpm and a temperature of 4oC until there was
a separation between the supernatant and the residue. The residue was filtered using Whatman No.41
filter paper and then dried in an oven at 60oC, prior to nutrient digestibility analysis.

2

334 Dasar Nutrisi Ruminansia (Edisi ke II)

a,b,c superscript means significantly different in a row (p<0.05)

3

335Dasar Nutrisi Ruminansia (Edisi ke II)

a,b,c superscript means significantly different in a row (p<0.05)

4

336 Dasar Nutrisi Ruminansia (Edisi ke II)