The Effect of Arrowroot Flour (Maranta Arudinaceae) to

Lactic Acid Bacteria Total and Feces Profile

in the Constipated Sprague Dawley Rats

Dewi Astuti1 , Lily Arsanti Lestari 2 and Diah Mulyawati Utari 3

Nutrition of Public Health, Faculty of Public Health, University of Indonesia
2
Health and Nutrition Departement, Faculty of Medicine, University of Gadjah Mada
3
Nutrition Department, Faculty of Public Health, University of Indonesia
1

Abstract. Constipation prevalence increases along lack of fruit and vegetable consumption in population 10 years
aged (93,6%). Nowadays, food products commonly use Fruktooligosakarida (FOS) and other Non-digestible
Oligosacharida (NDO) as prebiotic to promote Bifidobacteria-kind of Lactic Acid Bacteria (LAB) growth which has
beneficial for health. Arrowroot Flour (Maranta Arudinaceae) are rich in fiber, its fibers are fermented in the colon
and contribute to increase feces volume. This study aims to find out the implication of formulation diet with
arrowroot flour to LAB total in constipated rats through improved feces profile. This is experimental study with case
control design, conducted 30 days at central laboratory Gadjah Mada University. Starts with three days adaptation
phase, then 24 rats fed non fiber diet for seven days to make it constipated. After that, samples divided into 3 groups,
each group fed standar diet as Control group (CON), FOS diets (FOS), and Arrowroot Flour diets (AF). LAB total
measured from digesta samples, are taken on the last day study, while the feces profile (weight, volume, water
content, acidity) are analysed every three days. Data is processed by ANOVA and Kruskal Wallis test. It showed
LAB total of FOS is the highest, but theres not significant difference compared with AF (p=0,006). Theres
significant difference between LAB total of AF and CON (p= 0,023) and the feces profile (weight, water content,
acidity) of AF better than CON. These finding suggest, NDO from arrowroot flour should be considered as potential
prebiotics.
Keyword: Non digestible Oligoshacarida, Arrowroot, LAB total, feces profile, constipated

1. Introduction
Health problems that occur in many people nowadays is gastrointestinal disorder. One of
gastrointestinal disorder that can suffered by whole groups of age is constipation. Constipation is
defined as the the slow movement of stool through colon, caused of lasts longer fluid absorption so a
large number of dry and hard feces accumulate in the colon descenden (Guyton and Hall). Constipation
prevalence increases along lack of fruit and vegetable consumption in population 10 years aged
(93,6%), Riskesdas 2013. The characteristic of the food consumed play an important part. In this case,
food has a significant influence on feces and intestinal microflora (Mahan).
In javanese local wisdom believed that Arrowroot (Marantha Arudinaceae), known as Garut
Tubers has health benefits. One of its utilization is made of flour, which can be use as an alternative
ingredients for supplementary food for child under-two years and for people who are recovering from
illness (Purwantoro, 2000). Arrowroot flour (Marantha Arudinaceae) are rich in fiber, has a charateristics
as a antidiabetic component because of its non digestible fibers (Nursanti). The soluble fiber are
fermented in the colon, promote Lactic Acid Bacteria (LAB) growth and insoluble ones contributed to
increase feces volume trought water holding mechanism, so make feces consistency better then transit
time shorter (Marsono).
NDO are carbohydrates with a low molecular weight of intermediate in nature between
simple sugar and polysaccharides and has effect as prebiotic (Bathia) . Prebiotics are food ingredient
that couldnt be digested but it has positive effect on consumers health by stimulating the growth and
activities of several microflora in the colon selectively (kanthi). Fermentability of NDO will affect on
fecal bulk because of the number of bacteria in. Fructooligoscharida (FOS), galactooligoschararida
(GOS), inulin are kind of prebiotics.
In this study, health effect of prebiotics are focus on its capacity overcome constipated.
Nowadays, NDO as prebiotics have been widely used in commercial food products. Indonesia still

depend on import from another country due to the utilization from natural resources is not maximized.
So, the study aims to find out the functional effect of arrowroot flour with NDO as one of the ingredient
to modulate LAB and the laxative effect trought improved feces profile toward healthier condition in the
constipated rats.

2. Method
The current study had a randomize pre-post test control group design, and compared 3 group of
male Sprague Dawley rats (200-300 gr each) fed a standar diet (Control, n = 8), diet containing FOS 10%
(positive control, n= 8) and diet containing Arrowroot flour (AF), equal to 10% FOS (treatment, n=8).
FOS doses well tolerated in humans is 10 gr/day (Musatto). The dose converted in the rats feeding that
given ad-libitum. Diet feeding is made in a dose of 1000 grams, so FOS obtained the required weight of 9
grams. The addition of arrowroot flour in accordance with the principle of FOS are added as well. FOS is
a soluble fiber, so in the calculation of the addition of arrowroot flour into the feed calculated the water
soluble fiber content, so the figures obtained 179 grams.
The study starts with three days adaptation phase followed by conditioning phase when the 24
rats fed non fiber diet for seven days to make it constipated and then the intervention phase, 14 days. LAB
total measured from digesta samples, are taken on the last day study. Digesta sample taking by
determination rats with vertebral dislocation method, during surgery are taken the cecum to get digesta for
analysis. Analysis of LAB total using spread plated method by diluting 0.5 g digesta and dissolved in 4.5
ml of peptone water, then made dilutions serial (10 -1 10-7). The used data is from dilution serial
containing 30-300 Colony Forming Units (CFU).
The feces profile (weight, volume, water content, acidity) are analysed every three days and also
the body weight of the rats. Water content analysis is conducted using thermogravimetri method. The
acidity analysis by diluting the sample to 10 with distilled water, then measure the acidity with a pHmeter tool. The measurement of intake include weight and residual feed do everyday. All breeding phase
to rules outlined by Integrated Analysis and Reseach Laboratory University of Gadjah Mada.
3. Result and Discussion
Dietary intake of rats along this study are similiar. the results of statistic analysis is rats dietary
intake in all phases showed there is no significant difference (p>0.05). There is an increase and decrease
the weigh od rats in each phase but in general, test of analysis of variance showed that there is no
significant difference (p> 0.05). The dietary intake generaly no significant difference in adaptation phase
(p=0,670), conditioning phase (p=0,209), nor intervension phase (p= 0,105).
Lactic Acid Bacteria (LAB) Total : LAB total analysis from digesta samples because are more
sterile, after surgery due to direct evacuation and further processed according to the planting.

CON

FOS

AF

Based on these images can be observed that LAB total highest in FOS is 8.63 0.17 log CFU/g,
followed by AF of 8.55 0.11 log CFU/g and at least CON is 8,26 0.12 log CFU / g. LAB Total
meaningful difference exists between the FOS-CON (p= 0.006) and between AF-CON (p=0.023). While
between FOS-AF there is no significant differences in LAB total (p=0.532).
This result related with some literature review, daily food intake affects various types of bacteria
in the colon (Bathia). The number, types, and activity of bacteria in the colon was significantly affected
by supporting substrate such as redox potential, acidity, and oxygen availability (Sheila). NDO resistant to
enzymatic digestion and produce Short Chain Fatty Acid (SCFA), especially acetate and lactate with the
ratio 3:2 through colonic fermentation process (Ruberfroid). SCFA improve the acidity of caecum. The
acid environment is a good medium to stimulate the growth of beneficial colonic microflora. Some
research said that Bifidobacteria is one of the lactic acid bacteria that use NDO as its growth substrate.
Lactic Acid Bacteria is recognized has beneficial effect for digestion health (bielecka).

Feces Profile: Feces profile consist of weight, water content and acidity. Stool Weight: In all
groups, the stool weight decreased on the seventh day and declined sharply on the tenth day, it indicates
the rats in constipation. In intervention phase, the heaviest stool weigh is AF 1.13 0.308 g, followed
CON is 1.09 0.55 g and the lightest stool weight precisely FOS, namely 0.53 0.24 gr. Effects of
dietary fiber in increasing stool weight associated with increased bacterial cell mass in charge of
fermentation. On the other hand, fermentability fiber have a high water binding capacity (Marsono). The
stool weight of FOS is the lightest. According to qualitative observations, it because stool samples
consistency of FOS is the most mushy so causing not maximal of taking stool sampling. There is no
significant differences between AF-CON (p= 0,840) but AF stool weight heavier than CON. Judging
from its ability to increase stool output, preboitics a class of weak laxatives (Bathia). This shows that the
AF diet has the effect to increases stool weight but the effect is not strong enough.
Water content: The water content during the adaptation phase contained in these three groups is
not significant (p=0.624). The water content began to decline in the early conditioning phases. While on
the 10th and 12th days no faecal samples measured water content, because the effect of constipation. The
water content average during the intervention phase in the FOS remains the highest in the amount of
55.29 0.66 g%, AF is to 37.82 0.78 g% and CON is 35.25 0.62 gr%. Water content of FOS and AF
larger than CON. There is significant differences between AF-CON (p= 0,00). The fiber physical form
affect the absorption ability. Coarse fibers produces greater absorption than the fine ones ( Tensiska).
Particles fiber size affected water binding capacity. The larger particle size, the lower water solubility. It
caused fiber is fermented slowly in the colon then shorten the transit time. So it doesnt happen a lot of
water absorption in the colon (Musato).
Acidity: There is no significant acidity differences in adaptation (p = 0.595), and conditioning
phase (p = 0.079) between CON, FOS and AF. In intervention phase, there is significant acidity
differences between FOS-CON (p=0,00) and AF-CON (p=0,00). This result can be explain by Lisal, that
NDO is indigestible and can stimulate growth and/or metabolic activities of one or a group of colon
microflora. Colonic microflora will produce short chain fatty acid (SCFA) that use for energy
substrat for its growth and activities. The SCFA improve the feces acidity, stool mass and water
content (Marsono, Bielecka). This condition needed to tackling constipation condition.
4. Conclutions
Our experimental findings indicates that Arrowroot Flour diet has significantly affect LAB total
and feces profile especialy water content and acidity. In AF the stool weight is heavier than CON but its
not different statistically. Its show that Arrowroot Flour has a weak laxative effect although able to
modulate colon microflora growth. However concluded that Arrowroot Flour has a positive effect in
treating mild digestive disorder namely constipation.
Acknowlegdement
The Author would like to express sincere gratitude to Prof. Ratu Ayu Dewi Sartika, Dr.Diah Mulyawati
Utari

REFERENCES
1. Amenta, M., Cascasio, M.T., Di Fiore, P., Venturini, I. 2006. Diet and chronic constipation.
Benefits of oral supplementation with symbiotic zir fos (Bifidobacterium) longum W11 + FOS
Actilight). Acta Biomed, 77:157-162
2. Bathia, A., dan Rani, U. 2007. Prebiotics and health: Clinical Implications. Journal of
Clinical and Diagnostic Research, 6:546-554.
3. Bielecka, M., Biedrzycka, E., A., Juskiewicz, J., and Wrobleswka, M. 2002. Effect of nondigestible oligosaccharides on gut microecosystem in rats. Food Research Internastional,
35:139-144.

4. Bird, A.R.1999. Prebiotics: A role for dietary fibre and resistant starch. Asia Pacific. Journal
Clinical Nutrition, 8(suppl), S32-S36..
5. Guyton and Hall. 1997. Fisiologi Kedokteran. Edisi 9. Jakarta:EGC.
6. Kleessen, B., Syukra, B., Zunft, H-J., Blaut, M.1997. Effects if inulin and lactose on fecal
microflora, microbial activity, and bowel habit in elderly constipated persons. American
Journal of Clinical Nutrition, 65: (1) 397-402
7. Hana, 2007. Efek Serat Pangan dan Prebitotik (FOS ) yang Terkandung Pada Bubuk Pepaya
Meksiko Terhadap Profil Feses dan Total Mikroflora Digesta Tikus Sprague dawley. Program
Studi S1 Gizi Kesehatan UGM. Yogyakarta.
8. Marsono, Y. 2002. Indeks Glikemik Umbi-umbian. Agritech 22(1):13-16.
9. Marsono, Y. 2004. Serat Pangan Dalam Perspektif Ilmu Gizi. Pidato Pengukuhan Guru Besar.
Yogyakarta:UGM.
10. Marsono, Y.P. Wiyono, Zaki Utama. 2005. Indeks Glikemics Produk Olahan Garut
(Marantha arudinaceae) dan Uji Fungsionalitasnya pada Model Hewan Coba. Laporan
RUSNAS Diversifikasi Pangan Pokok Tahun 2005. Yogyakarta:UGM.
11. Mussatto, S.I., dan Mancilha. 2007. Non-digestible oligosaccharides: A review.
Carbohydrate Polymers, Vol 68, pp 587-597.
12. Ngatidjan, 2006. Metode Laboratorium dalam Toksikologi. Bagian Farmakologi dan
Toksikologi FK UGM. Yogyakarta
13. Nyman, M. 2002. Fermentation and bulking capacity of indigestibl carbohydrates: the case
of inulin and oligofructose. British Journal of Nutrition (2002), 87, Suppl. 2, S163S16
14. Niwa, T. et al., 2002. Effect of Dietary Fiber on Morphine-induced Constipastio in Rats,
Biosci. Biotechnol. Biochem., 66(6), 1233-1240
15. Ruberfroid, M.2007. Prebiotics: The conceps revisited. Journal of Nutrition,137: 830S
837S.
16. Salminen S, Bouley C, Ruault Boutron MC, S., C. Bouley2, M.-C. Boutron- Ruault3, J. H.
Cummings4*, A. Franck', G. R. Gibson6,E. Isolauri7, Roberfroid' and I. Rowland"et al.,1998.
Functional food science and gastrointestinal physiology and function. British Journal of
Nutrition 1998;80:S147-71
17. Smith B. V. Sc. John B, Mangkoewidjojo, S.1988. Pemeliharaan, Pembiakan dan
Penggunaan Hewan Percobaan di Daerah Tropis.Salemba 4, Jakarta: UI Press

18. Tensiska, Universitas Padjajaran. 2008. Serat Makanan. Diakses tanggal 6 Agustus 2008.
19. Van Loo, Cumming J., Nathalie Delzenne3, Hans Englyst2, Anne Franck1, Mark Hopkins2,
Nadine Kok3, George Macfarlane2, Dorothy Newton2, Michael Quigley2, Marcel
Roberfroid3,Trinette van Vliet4 and Ellen van den Heuvel4., 1999. Functional food
properties of non-digestible oligosaccharides:a consensus report from the ENDO project
(DGXII AIRII-CT94-1095). British Journal of Nutrition, (1999) 81, 121132.
20. Vuksan, V. 1999. A novel source of wheat fibre and protein: effect on fecal bulk and serum
lipid. American Journal of Clinical Nutrition, 69:226-300.
21. Wilhelm H. Holzapfel, Ulrich Schillinger. 2002. Introduction to pre- and probiotics: A
review. Food Research International, 35:109116.
Pembaharuan referensi
1. Riskesdas 2013
2. vitroXiao Wang1, Meiying Huang1, Fan Yang, Hanju Sun, Xianxuan Zhou, Ying
Guo,Xiaoli Wang, Manli Zhang, 2015. Rapeseed polysaccharides as prebiotics on
growth and acidifyingactivity of probiotics in vitro. vitroXiao. Carbohydrate Polymers
Journal. 125 (2015) 232240
3. Sheila Mara Sanches Lopesa, Mariane Grigio Franciscob, Bruna Higashia,Rafaela
Takako Ribeiro de Almeidac, Gabriela Krausovd, Eduardo Jorge Pilauc,Jos Eduardo
Gonc alvese, Regina Aparecida Correia Gonc alvesa,b,Arildo Jos Braz de
Oliveiraa,b, 2016. Chemical characterization and prebiotic activity offructooligosaccharides from Stevia rebaudiana (Bertoni) roots andin vitro adventitious root
cultures. Carbohydrate Polymers Journal. 152 (2016) 718725.
4. Shudong Hea, Xin Wanga, Yi Zhangb, Jing Wanga, Hanju Suna,, Junhui
Wanga,Xiaodong Caoa, Yongkang Yea 2016. Isolation and prebiotic activity of watersoluble polysaccharidesfractions from the bamboo shoots (Phyllostachys praecox),
Carbohydrate Polymers 151 (2016) 295304.

5.

Elena Rodrguez-Cabezas a, Desire Camuesco a, Beln Arribas a, Natividad GarridoMesa a,Mnica Comalada b, Elvira Bailn a, Margarita Cueto-Sola a, Pilar Utrilla a,
Eduardo Guerra-Hernndez c,Carlos Prez-Roca a, Julio Glvez a,*,d, Antonio Zarzuelo
a,d. 2010. The combination of fructooligosaccharides and resistant starch shows
prebiotic additive effects in rats. Clinical Nutrition 29 (2010) 832e839

6. Ka-Lung Lam,PeterChi-KeungCheungn. 2013. Non-digestible long chain beta-glucans

as novel prebiotics. B i o a c t i v e C a r b o h y d r a t e s a n d D i e t a r y F i b r e 2 ( 2
0 1 3 ) 4 5 6 4.

7. Peter Kurdi a, *, Chanida Hansawasdi. 2015. Assessment of the prebiotic potential of

oligosaccharide mixtures from rice bran and cassava pulp. LWT - Food Science and
Technology 63 (2015) 1288e1293.
8. Kanthi Kiran Kondepudi a,b,1, Padma Ambalam a,1, Ingrid Nilsson a, Torkel Wadstrm
a, sa Ljungh a,*2014. Prebiotic-non-digestible oligosaccharides preference of probiotic
bifidobacteria and antimicrobial activity against Clostridium difficile. Anaerobe 18 (2014)
489e497