Evaluation of Siger Rice from Waxy Cassava (Manihot Esculenta

Crantz) in Rat

Subeki

, Gusti Akhyar Ibrahim

, Tanto Pratondo Utomo

, Erwin Yuliadi

, Dewi Sartika

Department of Agricultural Product Technology in Faculty of Agriculture, Lampung University, Indonesia

Department of Mechanical Engineering in Faculty of Engineering, Lampung University, Indonesia

Department of Agrotechnology in Faculty of Agriculture, Lampung University, Indonesia

Keywords: Toxiological, Siger Rice, Waxy Cassava, Rat

Abstract: This study aimed to evaluate the toxicological potential of siger rice from waxy cassava in the rat. Siger rice

is a Lampung people's term "Beras Singkong Segar" which means artificial rice made from cassava. At first,

siger rice was made from native cassava containing high amylose. The study was conducted in a randomized

block design with ten replications. Siger rice is made from waxy cassava using a single screw extruder. Siger

rice fed to rats for 21 days had feed conversion of 4.26 and coefficient digestibility of 67.16%. Moreover, rat

fed siger rice had no negative haematological effect of packed cell volume, red blood cell and white blood

cell with each successive value of 24.21%, 4.17x106/mL, and 5.86x106/mL. Rats fed siger rice showed the

serum glutamate pyruvate transaminase, serum glutamate oxaloacetate transaminase, albumin, and bilirubin

with each successive value of 250.17 IU/L, 42.19 IU/L, 0.21 mg/dL, and 3.05 mg/dL no significant difference

with rat fed control. Further pathological investigation revealed that the liver had no necrotic lesion.

1 INTRODUCTION

The increased economic status of the community

caused by the increased food consumption exceeds

the body's needs. This condition causes nutritional

problems causing various degenerative diseases,

especially diabetes (Lathifah, 2017). Therefore,

alternative food ingredients need to be made to have

a good taste, low glycemic levels, low-calorie

content, and high levels of dietary fibre and bioactive

components. One alternative is siger rice.

Siger rice is the Lampung people's term "Beras

Singkong Segar", artificial rice made from cassava.

Siger rice was developed in Lampung to support food

diversification programs in reducing dependence on

rice (Henita, et al., 2018). Siger rice is made in the

form of grain, colour, and tastes like rice to be

accepted by the community and does not opposite

with the Indonesian eating habits. Siger rice, as

Lampung's fine local food since 2015, has been

instructed to become a food menu served in offices

and hotels in Lampung Province based on the

Governor's Instruction Lampung No:

521/1159/11.06/2015.

At first, siger rice is made from native cassava

flour containing high amylose content. In the

processing of siger rice, amylose gelatinized due to

the heating process. Therefore, the dough becomes

sticky and hard to be extruded into rice grains. High

amylose content in siger rice cause amylose to leak

out from the starch granules to bind an amount of

water during the cooking process and it will easily

rerelease the water until the rice becomes hard.

Therefore, this research will made siger rice from a

waxy cassava clone (Manihot esculenta Crantz)

containing high amylopectin and free amylose to

produce soft siger rice after being cold. The objective

of this research was to evaluate the toxicological

potentials of the siger rice from waxy cassava in rats.

This research was carried out in the Laboratory of

Quality Testing of Agricultural Product, Faculty of

Agriculture, Lampung University, and Laboratoirum

of Pathology of Regional Veterinary Investigation and

Testing of Lampung Province from May to October

2020.

The waxy cassava samples were collected from

the Integrated Field Laboratory of the Faculty of

Agriculture, Lampung University, Bandar Lampung.

The Wistar rat of 2 months old male were obtained

from the Regional III Center for Veterinary

Investigation and Testing of Lampung Province. Corn

oil, CMC, corn starch, vitamin mixtures, mineral

140

Subeki, ., Ibrahim, G., Utomo, T., Yuliadi, E. and Sartika, D.

Evaluation of Siger Rice from Waxy Cassava (Manihot esculenta Crantz) in Rat.

DOI: 10.5220/0010798400003317

In Proceedings of the 2nd International Conference on Science, Technology, and Environment (ICoSTE 2020) - Green Technology and Science to Face a New Century, pages 140-145

ISBN: 978-989-758-545-6

mixtures, casein, methionine, choline, formalin, xylol

solution, alcohol, paraffin, aqua dest, Harris's

Hematoxylin and Eosin. The tools used in this

research are extruder, grinding, hydraulic press,

hammer mill, blender, mixer, analytical balance,

sieve, filter, trash, heater, EDTA tube, Hematology

Analyzer, rat cage, and drinking bottle.

2 RESEARCH METHODS

The research was conducted in a complete

randomized block design with ten replications. The

study was conducted using 20 male Wistar rats

divided into control and siger rice diets. Each group

consists of 10 rats. Rats were treated with a

composition of casein, CMC, corn oil, corn starch,

siger rice, mineral mix, vitamin mix, DL-methionine,

choline, and water. Furthermore, keep the rats for 21

days and be fed ad libitum. Bartlet tested the

similarity of data, and Tuckey tested the addition of

data. Data were analyzed utilizing variation to obtain

the error estimator and significance test to determine

whether there was any difference between treatments.

The data analyzed by students t-test, signiﬁcance was

accepted at P<0.05 (Zar, 1984).

2.1 Implementation of Research

2.1.1 Cassava Flour

Cassava flour is made from waxy cassava (Subeki, et

al., 2016). The cassava tubers were peeled manually

with a knife, washed with tap water and crushed. The

crushed cassava was then pressed using a hydraulic

press. The pressed pulp was later subjected to the

oven at 60

C and ground into cassava ﬂour.

2.1.2 Siger Rice

Siger rice made from cassava four according to the

method of Subeki (Subeki et al., 2016). The making

of siger rice is done by mixing cassava flour with the

addition of 30% water. The dough ingredients are

mixed with a mixer and then steamed for 30 minutes.

The material is then put in a single screw of extruder

machine at 45 rpm rotation, 40 rpm cutting blade

rotation, with dyes a siger rice hole of ellipse with 6

mm long and 2 mm thick to obtain siger rice grains.

Siger rice was then dried in an oven at 60

C until the

moisture of less than 13%.

2.1.3 Sample Analysis

Analysis of protein content was determined by the

micro-Kjeldahl method (AOAC, 1990). Siger rice 0.5

g is hydrolyzed with 10 mL H

using

CuSO

.5H

O catalyst until the solution becomes

grey-white. The hydrolyzed filtrate 10 mL were

distilled with 15 mL 40% sodium hydroxide for 10

minutes into 20 mL 2% boric acid containing three

drops of indicator solution (a mixture of 0.018 g

bromocresol green and 0.016 g methyl red in 100 mL

water). The distillate was then titrated with 0.01 M

HCl. The nitrogen (N) content is calculated from the

titer and crude protein is obtained by multiplying the

nitrogen content by the conversion factor Nx6.25.

The blank determination is carried out

simultaneously. Analysis of moisture, ash, crude

fiber, fat, and carbohydrates using the AOAC method

(AOAC, 1990).

2.1.4 Nutritional Quality

The analysis of the nutritional quality of siger rice in

Wistar rats divided into two groups. Rat in each group

kept in individual wire cages at room temperature.

The feed was formulated by replacing cornflour with

siger rice (Autor, 1993). Feed intake, daily weight

gain, feed efficiency and digestibility were measured

during the experiment. At the end of the 21-day trial

period, all rats were killed by asphyxiation and blood

was taken by heart puncture, and the serum was then

stored at -20

C until it being analyzed. The liver and

kidneys remove for further examination. The feed

composition of basal siger rice ﬂour in the rat can be

seen in Table 1.

Table 1. Feed composition of basal siger rice ﬂour

(g/1000 g)

Sampel

Diet

Standard (AIN-76) Siger rice

Casein 200 146.4

CMC 50 43.35

Corn oil 180 176.65

Corn starch 539 130.25

er rice - 500

Mineral mix 135 133.15

Vitamin mix 10 10

dl Methionine 3 3

Colin 1 1

Wate

25.8 0

Total 1043.8 1043.8

Evaluation of Siger Rice from Waxy Cassava (Manihot esculenta Crantz) in Rat

141

2.1.5 Haematology Tests

Haematological tests on the number of packed cell

volume (PCV), red blood cells (RBC) and white

blood cells (WBC) were conducted using the

conventional method (Aning, 1998). Biochemical

tests on albumin, bilirubin, Glutamate Oxaloacetate

Transaminase (GOT), and Glutamate Pyruvate

Transaminase (GPT) were carried out by

conventional methods (Mokadi, et al., 1989).

2.1.6 Histopathological Tests

The histopathological test was carried out by taking

the liver and kidneys of each rat and then stored in a

4% formalin-saline solution. After dehydration and

embedding in parafﬁn wax, 5-mm sections were cut

and stained for examination in the microscope. The

internal organs were inspected for morphological

lesions.

3 RESULTS AND DISCUSSION

The results showed that the bodyweight of the rats

increased every day in the group of mice were fed

control and siger rice diets. The increase in body

weight indicates good feed conversion efficiency and

overall performance. It is a good indicator that the

diet fed to mice has a high nutritional quality (Van,

1999). Table 2 shows that the control diet-fed mice

showed higher body weight gain than the siger rice

diet-fed mice (Figure 1). The higher growth rate in

mice fed the control diet was due to the higher feed

intake (Aning, et al., 1998). The mice fed the control

diet had a higher feed intake than those fed the siger

rice diet. It shows the amount of feed consumption

plays a vital role in the growth of the rat (Aning, et

al., 1998).

Table 2. Nutrient utilization and digestibility of rats fed

siger rice ﬂour

Sample Control Siger Rice

Daily weight gain (g) 2.72 + 0.17

2.05 + 0.33

Daily feed intake (g) 9.53 + 0.12

8.74 + 0.83

Feed:Gain ratio 3.50 + 0.71

4.26 + 0.40

Daily excretion (g) 3.12 + 0.22

2.87 + 0.31

Digested amount (g) 6.41 + 0.73

5.87 + 0.88

Coeficient digestibility

(%)

67.26 + 5.25

67.16 + 6.63

Dry matter digestibility

(%)

68.56 + 5.38

66.32 + 6.95

Values are mean + S.E (n =3). Means with the same

superscript letter(s) along the same row are not

signiﬁcantly different (P>0.05).

Figure 1. Growth curve of rats fed siger rice

The rats fed the control diet had a lower ratio, than

those fed by the siger rice diet. It shows that the low

growth in rats fed the siger rice is not due to poor

nutrient quality but the low amount of feed intake

(Aning, et al., 1998). However, rats efficiently

converted the little feed consumed into weight gain

(Aning, et al., 1998).

Digestibility results of siger rice are shown in

Table 2. There was no significant difference in the

digestibility coefficient of control and siger rice. The

value of this digestibility coefficient is higher than

cassava processed by fermentation but lower than

cooked casein by 91.2% (Bressani, 1999). It shows

that the protein in the feed is very easy to digest. This

high digestibility indicates a reduction in tannins

during cassava processing.

The results showed no significant difference

between dry matter digestibility in the control and

siger rice diet. The dry matter digestibility of siger

rice is lower than that of raw sweet potatoes (85.4%)

and cooked sweet potatoes (90.4%) (Canopy,

Tamiya, 1977). These results indicate that the siger

rice-based diet has high digestibility. Therefore,

processed siger rice by extruders can improve the

nutritional quality of the product.

Table 3 shows the levels of GPT, GOT, bilirubin,

and albumin in the serum of rats fed control and siger

rice. The results showed no significant difference

between GOT levels in the control (247.11 IU/l) and

siger rice (250.17 IU/l) diets. There was no increase in

GOT in rat fed siger rice due to no secretion of toxins

during the processing of siger rice. Aflatoxin is a toxic

compound that can cause liver damage (Oboh, et al.,

0 6 1218

Weightgain(g)

Day

Contr

Siger

rice

ICoSTE 2020 - the International Conference on Science, Technology, and Environment (ICoSTE)

142

2000). Increased blood GOT valuesare attributed to

liver damage and possible heart damage [(American

Liver Foundation, 1995), (David, 1999)]. There was

no significant difference between the GPT levels in

the blood of rats fed control (39.14 IU/l) and siger rice

(42.19 IU/l) diets. There was no increase in the GPT

value in rat fed siger rice, indicating that cassava

processing did not produce toxins (Oboh, et al., 2000).

GPT and GOT are enzymes located in liver cells

that can leak into the blood circulation when liver

cells are injured. GPT levels are a specific indicator

of liver inflammation. GOT levels can be increased in

a human who has heart disease [(American Liver

Foundation, 1995), (American Liver Foundation,

1997), (David, 1999)]. Acute liver disease such as

hepatitis shows very high GPT and GOT levels of

more than 1000 IU/l (American Liver Foundation,

1997). GPT and GOT levels are highly dependent on

the range of liver disease [(American Liver

Foundation, 1995), (American Liver Foundation,

1997), (David, 1999)].

Table 3. Serum chemistry and haematological evaluation of

rats fed siger rice flour

Sample Control Siger rice

GOT (IU/L) 247.11 + 6.13

250.17 + 5.11

GPT (IU/L) 39.14 + 4.25

42.19 + 2.06

Albumin (mg/dL) 0.19 + 0.14

0.21 + 0.21

Bilirubin (mg/dL) 3.26 + 0.17

3.05 + 0.26

PCV (%) 23.68 + 0.23

24.21 + 2.16

RBC (10

/mL) 3.87 + 0.18

4.17 + 0.15

WBC (10

/mL) 9.91 + 0.91

5.86 + 1.03

Values are mean + S.E (n =10). Means with the same

superscript letter along the same row are not

signiﬁcantly different (P>0.05).

There was no signiﬁcant change (P>0.05) in serum

bilirubin content of the rats fed siger rice ﬂour diet

compared with that of the control diet. This low level

of bilirubin in the serum of the rats fed diets containing

siger rice ﬂour is an indication that the feed did not

cause haemolysis of red blood cells since bilirubin is

formed primarily from the breakdown heme

[(Lehninger, 1987), (American Liver Foundation,

1997)].

The results showed that the serum bilirubin content

of rat fed siger rice of 3.05 mg/dL was not

significantly different from the control 3.26 mg/dL.

The low bilirubin content in the serum of rat fed siger

rice indicates that administration of siger rice does not

cause hemolysis of red blood cells. Bilirubin is a

compound formed from the breakdown of heme

[(Lehninger, 1987), (American Liver Foundation,

1997)].

Rats fed siger rice showed no significant

difference in albumin level (0.21 mg / dL) from that

of control (0.19 mg/dL). These results indicate that

the administration of cassava inhibits albumin

biosynthesis in the liver. Albumin is produced

primarily in the liver as an indicator of liver function

[(American Liver Foundation, 1995), (American

Liver Foundation, 1997), (David, 1999)].

The haematology test of rats fed siger rice showed

the levels of PCV (24.21%) and RBC (4.17x10

/mL)

were not significantly different from the levels of

PCV (23.68%) and RBC (3.87x10

/mL) control.

However, the rat fed siger rice showed significantly

lower WBC levels (5.86x10

/mL) than the control

WBC 9.91x10

/mL. The low white blood cell count

in siger rice is due to the bioactive components in

cassava, which can inhibit the growth of pathogenic

microorganisms. The PCV, RBC, and WBC analysis

results are in line with reported in rat fed sorghum

(Aning, et al., 1998). These results also align with

processed cassava products that do not show

haematological effects (Autor, 1993). The

histopathological test of the kidney and liver of the

rats fed siger rice showed that the diet did not cause

any damage to the organ. The pathological studies

had shown clearly that the siger rice diet could cause

damage to the liver, as earlier indicated by its low

serum GPT and GOT activities [(American Liver

Foundation, 1995), (American Liver Foundation,

1997), (David, 1999), (Younossi, Mehta, 1998)].

Further pathological investigation showed that the

liver had no necrotic lesion. Cell appearance of

kidney and liver can be seen at Figure 2 and 3.

(a)

(b)

Figure 2. Histological liver of HE stained rat with

magnification 100x. (a) control and (b) siger rice

Evaluation of Siger Rice from Waxy Cassava (Manihot esculenta Crantz) in Rat

143

(a)

(b)

Figure 3. Histological kidney of HE stained rat with

magnification 100x. (a) control and (b) siger rice

4 CONCLUSIONS

Siger rice fed to rat for 21 days had feed conversion

of 4.26 and coefficient digestibility of 67.16%.

Moreover, rat fed siger rice had no negative

haematological effect of packed cell volume, red

blood cell and white blood cell with each successive

value of 24.21%, 4.17x10

/mL, and 5.86x10

/mL.

Rats fed siger rice showed the serum glutamate

pyruvate transaminase, serum glutamate oxaloacetate

transaminase, albumin, and bilirubin with each

successive value of 250.17 IU/L, 42.19 IU/L, 0.21

mg/dL, and 3.05 mg/dL no significant difference with

rat fed control. Further pathological investigation

revealed that the liver had no necrotic lesion.

ACKNOWLEDGMENTS

The authors are grateful to the Ministry of Research,

Technology, and Higher Education for funding

through Wold Class Research grants 2020.

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