Chapter 4 PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA

This chapter presents the data gathered through actual observation and the changes that happened to the blood vessels of the Duck eggs incubated for 10 days.

4.1 Results of the Phytochemical Screening

The results obtained from the Phytochemical screening shows the Bagras and Chico leaves secondary metabolites. Table 4.1 shows the results. Secondary Bagrass Metabolites Alkaloids Saponins Flavonoids Unsaturated Sterols Leucoanthocyanins Tannins Positive Positive Negative Negative Positive Positive Chico Positive Negative Positive Positive Positive Positive

The table shows the positive metabolites as well as negative metabolites in the Bagrass plant and the Chico Plant. Alkaloids, Saponins, Leucoanthocyanins, and Tannins are all positive metabolites of Bagras extract; while Flavonoids and Unsaturated Sterols are the negative metabolites of the said plant. The Chico extracts are positive

for Alkaloids, Leucoanthocyanins, Tannins, Flavonoids and Unsaturated Sterols. On the hand, the extract is negative to Saponins.

4.2 Results of the Anti-Angiogenic Screening

Treatment Chico Bagras Combined Simvastatin Water

1 15 15 14 20 32

REPLICATIONS 2 18 18 18 17 30

3 13 18 15 12 29

TOTAL 46 51 47 49 91

MEAN 15.33 17 15.67 16.33 30.33

The table shows the number of blood vessels in the three replications of each treatment. Chico extract showed the least mean with 15.33 blood vessels and the negative control shows the greatest mean with 30.33 blood vessels. Compared to the positive control, the table shows that the combined extract and the Chico extract are better in decreasing the number of blood vessels.

Anova:Anti-Angiogenic Property
n 15

Treatments Chico Bagras Combined Simvastatin Water

n 3 3 3 3 3

Mean 15.3 17.0 15.7 16.3 30.3

SE 1.45 1.00 1.20 2.33 0.88

Pooled SE 1.47 1.47 1.47 1.47 1.47

SD 2.5 1.7 2.1 4.0 1.5

Source of variation Treatments Residual Total LSD Contrast Chico v Bagras Chico v Combined Chico v Simvastatin Chico v Water Bagras v Combined Bagras v Simvastatin Bagras v Water Combined v Simvastatin Combined v Water Simvastatin v Water

Sum squares 492.3 64.7 556.9

DF 4 10 14

Mean square 123.1 6.5

F statistic 19.03

p 0.0001

Difference -1.7 -0.3 -1.0 -15.0 1.3 0.7 -13.3 -0.7 -14.7 -14.0

95% CI -6.3 to 3.0 -5.0 to 4.3 -5.6 to 3.6 -19.6 to -10.4 -3.3 to 6.0 -4.0 -18.0 -5.3 -19.3 -18.6 to 5.3 to -8.7 to 4.0 to -10.0 to -9.4

(significant)

(significant)

(significant) (significant)

Analysis of Variance revealed that the F statistics of 19.03 which is greater than the pooled value of 0.0001 which dictated for the rejection of the null hypothesis; that the mean difference between treatments has no difference. Moreover, the Least Significance Difference Test showed that the average number of membrane formations in each treatment significantly differ only it the treated and the positive control group (Simvastatin) is compared to the negative control group (water). Evidently the treated group showed no differences in the mean membrane formation when compared to the positive group (Simvastatin). This means, that Bagras and Chico

extract or its combination exhibited equal effect on the membrane formation in the experimental duck egg. Thus, Cogon and Bagras or its combined extract has anti-angiogenic property. Graph I: Anti-Angiogenic Property or Bagras
16 14 12 10 8 6 4 2 0 T0 T1 T2 T3 R1 R2 R3

The graph clearly states that the replications of even the same treatment are varied. The replication with the lowest number of veins is R2 of T1, standing out from the measurements of other replication because of its large gap it made from others. The replication which has the most number of veins is R2 of T0.

4.3 Results of the Kirby-Bauer Testing

This subchapter discusses the data gathered after the KirbyBauer Testing. The results showed that the extracts did have antibacterial properties. Escherichia coli and Staphylococcus aureus both showed positive results.

4.3.1 Zone of Inhibition in Staphylococcus aureus

REPLICATIONS Treatment Norfloxacin Bagras Chico Combined 1(mm) 29.80 29.60 28.75 18.25 2(mm) 30.25 29.00 29.25 15.75 3(mm) 30.15 28.25 25.15 16.50 TOTAL(m m) 90.2 86.85 83.15 50.5 MEAN(mm ) 30.07 28.95 27.72 16.83

Table 4.3.1 shows the results of the Kirby Bauer Testing on Staphylococcus aureus is positive in all treatments. The control still showed the greater mean with 90.2 followed by Bagras with a mean of 86.85, next is Chico with a mean of 83.15, and lastly, combined extract showed the least mean with only 50.5.

ANOVA: Growth Inhibition of Staphylococcus aureus (Gram Positive Bacteria)

12

<Variable> by Treatments Norfloxacin Bagras Chico Combined

n 3 3 3 3

Mean 30.067 28.950 27.717 16.833

SE 0.1364 0.3905 1.2914 0.7407

Pooled SE 0.7726 0.7726 0.7726 0.7726

SD 0.236 0.676 2.237 1.283

Source of variation Treatments Residual Total

Sum squares 336.504 14.325 350.829

DF 3 8 11

Mean square 112.168 1.791

F statistic 62.64

P <0.0001

LSD Contrast Bagras v Norfloxacin Chico v Norfloxacin Combined v Norfloxacin Difference -1.117 -2.350 -13.233 95% CI -3.636 to 1.403 -4.870 to 0.170 -15.753 to -10.714
(significant)

Analysis of Variance revealed that the F statistics of 62.64 which is greater than the pooled value of lesser than 0.0001 which dictated for the rejection of the null hypothesis; that the mean difference between treatments has no significant difference. Moreover, the Least Significance Difference Test showed that the mean zone of inhibition exhibited by the combined extract with that of Norfloxacin against

Staphylococcus aureus has significant difference while the individual extract showed no sigficant difference in the mean zone of inhibition against the S. aureus. This means, that Bagras and cogon extract has antibacterial property against gram positive bacteria. Moreover, there is no synergistic anti-bacterial property of the said extracts. This may seem that Bagras and Chico leaves extract has antagonistic effect.

Graph II: Zones of Inhibition of Bagras on Escherichia coli

35 30 25 20 15 10 5 0 Norfloxacin Bag ras C h ico C om b in ed R1 R2 R3

The graph clearly states that the replications of even the same treatment are varied. The replication with the highest zone of

inhibition is R2 of Norfloxacin, standing out from the measurements of other replication because of the gap it made from others. The

replication which has the smallest zone of inhibition is R 2 of Combined extract.

Zone of Inhibition in mm

4.3.2

Results for Escherichia coli

REPLICATIONS Treatment Norfloxacin Bagras Chico Combined 1(mm) 21.75 17.75 22.50 20.75 2(mm) 23.15 14.25 21.50 22.45 3(mm) 25.10 18.25 23.25 22.75 TOTAL(mm) 70 50.25 67.25 65.95 MEAN(mm) 23.33 16.75 22.42 21.98

Table 4.3.1 shows the results of the Kirby Bauer Testing on Escherichia coli is positive in all treatments. The control still showed the greater mean with 70 followed by Chico with a mean of 67.25, next is Combined with a mean of 65.95, and lastly, Bagras extract showed the least mean with only 50.25.

ANOVA : Gram Negative Bacteria

n 12

Treatments Norfloxacin Bagras Chico Combined

n 3 3 3 3

Mean 30.067 28.950 27.717 16.833

SE 0.1364 0.3905 1.2914 0.7407

Pooled SE 0.7726 0.7726 0.7726 0.7726

SD 0.236 0.676 2.237 1.283

Source of variation Treatments Residual Total

Sum squares 336.504 14.325 350.829

DF 3 8 11

Mean square 112.168 1.791

F statistic 62.64

P <0.0001

LSD Contrast Bagras v Norfloxacin Chico v Norfloxacin Combined v Norfloxacin Difference -1.117 -2.350 -13.233 95% CI -3.636 to 1.403 -4.870 to 0.170 -15.753 to -10.714
(significant)

Analysis of Variance revealed that the F statistics of 62.64 which is greater than the pooled value of lesser than 0.0001 which dictated for the rejection of the null hypothesis; that the mean difference between treatments has no significant difference. Moreover, the Least Significance Difference Test showed that the mean zone of inhibition

exhibited by the combined extract with that of Norfloxacin against Staphylococcus aureus has significant difference while the individual extract showed no sigficant difference in the mean zone of inhibition against the S. aureus. This means, that Bagras and cogon extract has antibacterial property against gram positive bacteria. Moreover, there is no synergistic anti-bacterial property of the said extracts. This may seem that Bagras and Chico leaves extract has antagonistic effect.

Graph III: Zones of Inhibition of Bagras on Staphylococcus aureus

30 25 20 15 10 5 0 Norfloxa c in B a g ra s Chic o Com bined R 1 R 2 R 3

The graph clearly states that the replications of even the same treatment are varied. The replication with the highest zone of

inhibition is R3 of Norfloxacin, standing out from the measurements of other replication because of the gap it made from others. The

replication which has the smallest zone of inhibition is R2 of Bagras extract.

Zone of Inhibition in mm