GROUP 5

Group Members:
Farah Najwa bt Zainal Abidin Noor Nabila bt Mohd Radzi Mohammad Syafiq Iqbal b Mohammad Helmi Nurul Syahirah bt Ahmad Noor Nazira bt Kadir Halimatun Sadiah bt Jefri Norfaizah bt Mohd Yunus Nurul Ashikin bt Mohamad Wasis uk24868 uk25450 uk25620 uk25083 uk25156 uk25283 uk24773 uk25316

For your considerations

well done BRAVO Group 5

Split-Plot Design (SPD)

Question
Peuyeum (fermented cassava) is made by bacterial fermentation of yeast. Most of the bacteria are purposefully added; these are the starter cultures. Some wild bacteria are also present in Peuyeum; these are nonstarter bacteria. This experiment explores how intentionally-added nonstarter bacteria affect Peuyeum quality. We use four strains of nonstarter bacteria: R1, R2, R3 and R4 as Subplots. Our Main Plot treatments will be control (A: local cassava); B: Malay cassava; C: Indonesian cassava . After 15 days of ripening, each Peuyeum is measured for total free amino acids (a measure of bacterial activity related to Peuyeum quality). Responses are given in Table 5.

Experimental terms
Experimental unit : cassava Main plot : Peuyeum (fermented cassava)

- A : Thailand Cassava - B : Malay Cassava - C : Indonesian Cassava

Subplot : nonstarter bacteria

- R1 : Lactobacillus nagelli - R2 : Lactobacillus harbinensis - R3 : Lactobacillus buchneri - R4 : Lactococcus lactis

Experimental for block

In the fermentation handling we used the types of

container (block) to store the peuyeum after added the nonstarter bacteria on cassava.
The 3 blocks are :

- block 1 ( clay pot) - block 2 ( aluminium pot) - block 3 ( plastic container)

Experimental Design
Main plot : The types of

Peuyeum (fermented

cassava) such as A : Thailand Cassava, B : Malay

Cassava and C : Indonesian Cassava .
Subplots : The nonstarter bacteria can classified

by 4 types of bacteria which are R1: Lactobacillus nagelli, R2 : Lactobacillus harbinensis, R3 : Lactobacillus buchneri and R4 : Lactococcus lactis.

Table 5. The total free amino acids (mg/g)

Block AR1 AR2 AR3 AR4 BR1 BR2 BR3 BR4 CR1 CR2 CR3 CR4 1 5.1 6.1 9.0 17.5 8.6 10.5 16.5 19.3 8.9 12.0 11.0 19.6

Block AR1 AR2 AR3 AR4 BR1 BR2 BR3 BR4 CR1 CR2 CR3 CR4 2 9.5 10.6 8.9 17.5 5.5 8.2 17.4 17.3 8.0 17.8 13.0 18.6

Block AR1 AR2 AR3 AR4 BR1 BR2 BR3 BR4 CR1 CR2 CR3 CR4 3 7.1 6.5 9.0 17.4 7.5 8.9 15.5 15.3 8.7 17.8 18.0 18.6

Interaction term

AR1 : Thailand Cassava x Lactobacillus nagelli AR2 : Thailand Cassava x Lactobacillus harbinensis AR3 : Thailand Cassava x Lactobacillus buchneri AR4 : Thailand Cassava x Lactococcus lactis BR1 : Malay Cassava x Lactobacillus nagelli BR2 : Malay Cassava x Lactobacillus harbinensis BR3 : Malay Cassava x Lactobacillus buchneri BR4 : Malay Cassava x Lactococcus lactis CR1 : Indonesian Cassava x Lactobacillus nagelli CR2 : Indonesian Cassava x Lactobacillus harbinensis CR3 : Indonesian Cassava x Lactobacillus buchneri CR4 : Indonesian Cassava x Lactococcus lactis

Randomized)
Main plot Fermented Cassava A Fermented Cassava AR CR CR CR AR AR 4 AR 2 3 CR 4 1 3 17. 2 12. 11. 1 19. 5.1 9.0 5 6.1 0 0 8.9 6 CFermented Cassava B BR BR BR BR 3 2 4 Sub plot 1 16. 10. 19. 8.6 5 5 3

Block 1

Block 2

Fermented Cassava B Fermented Cassava A Fermented Cassava C BR BR AR AR CR CR CR 3 BR BR 4 4 2 AR AR 3 2 4 CR 17. 1 2 17. 17. 10. 1 3 13. 17. 18. 1 4 5.5 8.2 3 5 6 9.5 8.9 0 8 6 8.0 Fermented Cassava C Fermented Cassava B Fermented Cassava A CR CR CR BR BR AR 2 4 CR 3 3 4 BR BR AR AR AR 4 17. 18. 1 18. 15. 15. 2 1 2 1 3 17. 8 6 8.7 0 5 3 8.9 7.5 6.5 7.1 9.0 4

Block 3

Entering Data to SPSS

Step 1: Enter the Experiment Data

Step 2 : Enter the value labels (Block)

Step 3: Analyze General Linear Model Univariate

Step 4: Custom Model for Block ANOVA

Anova analysis: From the table obtained sig. of treatment and block Block : sig.= 0.00 Treatment :sig.= 0.00 Significant level = 0.05

ANOVA for Block

Block Analysis data

If significance value > 0.05, H0 is accepted If significance value < 0.05, H0 is rejected
From the table, the significant value of block is:

0. 819> 0.05. The value is not significant, means that the block has no effect to the experiment. So, we can ignore the block ( types of container: clay pot, aluminium pot and plastic container) in the experiment because it will not influence the nonstater bacteria and cassava to produce the production of total free amino acid in the peuyeum. Thus, we continue (Full-Factorial) for interaction between types of cassava and non stater bacteria.

Model full Factorial

Data Output ANOVA Table

Analysis Data
If significance value > 0.05, H0 is received If significance value < 0.05, H0 is rejected
Significance value: Bacteria : 0.00 (<0.05) Cassava : 0.00 (<0.05) Bacteria*Cassava : 0.02 (<0.05)

Analysis For Bacteria

The sig value is 0.00 (<0.05), so H0 is rejected. In other words, H1 is receive as the value is VERY SIGNIFICANT. This means that the bacteria have effects towards total free amino acid production in cassava.

Analysis for Cassava

The sig value is 0.000 (<0.05), so H0 is rejected. In other words, H1 is receive as the value is VERY SIGNIFICANT. This means that the types of fermented cassava have effects towards total free amino acid production.

Analysis For Bacteria* Cassava

The sig value is 0.002 (<0.05), so H0 is rejected. In other words, H1 is receive as the value is VERY SIGNIFICANT. This means that the bacteria have effects towards total free amino acid production in cassava.

The Step to Determining the Best Interaction Since the interaction of bacteria and types of fermented cassava to total free amino acid production is very significant, so we continue to Completely Randomized Design (CRD) and Post Hoc Test in order to find out the best interaction treatment

Completely Randomized Design (CRD)

CRD Data taken from above SPD

Treatment Interaction AR1 AR2 AR3 AR4 Replications (Block) 1 (A)
5.1 6.1 9.0 17.5

2 (B) 9.5 10.6 8.9 17.5

3 (C) 7.1 6.5 9.0 17.4

BR1
BR2 BR3 BR4

8.6
10.5 16.5 19.3

5.5
8.2 17.4 17.3

7.5
8.9 15.5 15.3

CR1
CR2 CR3 CR4

8.9
12.0 11.0 19.6

8.0
17.8 13.0 18.6

8.7
17.8 18.0 18.6

Treatment Interaction
(1) AR1 : Thailand Cassava x Lactobacillus nagelli (2) AR2 : Thailand Cassava x Lactobacillus harbinensis (3) AR3 : Thailand Cassava x Lactobacillus buchneri (4) AR4 : Thailand Cassava x Lactococcus lactis (5) BR1 : Malay Cassava x Lactobacillus nagelli (6) BR2 : Malay Cassava x Lactobacillus harbinensis (7) BR3 : Malay Cassava x Lactobacillus buchneri (8) BR4 : Malay Cassava x Lactococcus lactis (9) CR1 : Indonesian Cassava x Lactobacillus nagelli (10) CR2 : Indonesian Cassava x Lactobacillus harbinensis (11) CR3 : Indonesian Cassava x Lactobacillus buchneri (12) CR4 : Indonesian Cassava x Lactococcus lactis

Field Layout of Experimental Designs (Before Randomized)

AR

AS

AP

AQ

BR

BS

BP

BQ

CR

CS

CP

CQ

Replication (3 times: A, B, C, )

Field Layout of Experimental Designs (After Randomized)

Layout of experimental unit for field observations

5A Replicatio n (3 times: A, B, C,

1C

8A

4B

7C

10A 12 B 11B 2C

10C 3A

7B

4C

1B

2B

11A 8C

6A

2A

5B

5C

10B 8B

11C

12C 7A

3C

12A 9C

4A

9B

6C

3B

1A

6B

9A

SPSS for CRD

The Procedures of Experimental Designs

Step 1- Entering the data into SPSS

Step 2: Enter the value labels

Step 3: Analyzec compare means one way ANOVA

SPSS Output

Test of Homogenity

Not homogenous because 0.004 < 0.05.

Test of Homogenity
Significant correction = 0.05 If the significant obtain > , so the variance of each samples is homogenous.
If the significant obtain < , so the variance of each sample is not homogenous.

ANOVA Table

Hypothesis Testing Analysis by Significant Value

Ho = 1=2=3=4=5=6=7=8=9=10=11=12 H1 = Not all are equal Significant correction = 0.05 If the significant obtained > , so the variance of each samples is homogenous. If the significant obtained < , so the variance of each samples is not homogenous. Conclusion : Because of the significant value is 0.000, that means the sig. value < 0.05 and the value is very significant (< 0.01), so from all interaction, at least there is one interaction

Continue to Post hoc test

Output of post hoc

Cont. Output of post hoc

Cont. Output of post hoc

Cont. Output of post hoc

Cont. Output of post hoc

Cont. Output of post hoc

Result and Discussion Analysis

From post hoc table, the sig. value for (AR4)

Thailand cassava and lactococcus lactis, (BR3) Malay cassava and lactobacillus buchneri, (BR4) Malay cassava and lactococcus lactis , and (CR4) Indonesian Cassava and Lactococcus lactis. are (0.00 <0.05) and the sig. value for Indonesian Cassava and lactobacillus harbinensis is (0.01<0.05).

Homogenous

Means for groups in homogenous subset are displayed

Homogenous Subset
Treatment Interaction AR1 (Thailand Cassava x Lactobacillus nagelli) AR2 (Thailand Cassava x Lactobacillus harbinensis) AR3 (Thailand Cassava x Lactobacillus buchneri AR4 (Thailand Cassava x Lactococcus lactis) N 3 3 3 3 Mean 7.200 2.2030 a 7.233 2.4906 a 7.733 0.577 a 8.533 0.577 ab

BR1 (Malay Cassava x Lactobacillus nagelli)

BR2 (Malay Cassava x Lactobacillus harbinensis) BR3 (Malay Cassava x Lactobacillus buchneri) BR4 (Malay Cassava x Lactococcus lactis)

3
3 3 3

8.967 1.5716 ab
9.200 1.1790 ab 14.000 0.9504 bc 15.867 2.000 c

Means bearing the same superscript within the same CR1 (Indonesian Cassava x Lactobacillus different 3 0.4726 c column are not significantly at16.467 5% level nagelli) (p<0.05)

Means Graph of Interactions

Although we can see the best interaction from

homogenous subset table but we also can use another method that is graph which is taken from the SPSS output. From the graph, we can also observed which one is the best interaction by analyze the mean of mean total free amino acid against the interaction (by using Graph or Bar Chart).

Output SPSS also shows Means Graph (Mean of amino acid against the interaction)

Interaction

Conclusion
Based on Table;

The best interaction in formation of peuyeum is CR4 (Indonesian Cassava and lactobacillus buchneri) because the nonstarter bacteria of Lactobacillus buchneri is a good inducer of the optimum production of the free amino acid in cassava and induce fermentation of carbohydrate content in the tuber to be sugar for just 2-3 days. An optimum production of free amino acids in fermented cassava will gives the highest quality of peuyeum with the best flavour development, which are not too ripe that gives too sour taste. Besides, It will improve the production of lactic acid which is one of the types of amino acid that affected the development of flavour, taste, and texture. Cassava are characterized by higher