Good nutrition is a key to a healthy lifestyle.

• US Department of Agriculture (2015)

Mitchell (1997) & Dossey, (2008):

• Every day, an average of two to three new

synthetic chemicals are released to the
environment

• On the 21st century 70,000 different

synthetic chemicals on the global market
 “Pectobacterium carotovorum is an ubiquitous plant
pathogen that causes bacterial soft rot on nearly every type of
fruit and vegetable produced in tropical and temperate
regions.”
Charkowski, Condemine, Expert, Hayes, Toth et al., (2012).
 According to Elphinestone (2010), Pectobacterium
carotovorum is the major causative agent of bacterial soft rot.

The presence of bacterial soft rot increases the growth

of Salmonella Typhimurium and Eschericia coli (Bais &
Sherrier, 2015)
 (Salunke & Kadam, 2005 ; Simeon and Abubakar, 2014)
Borax is the best synthetic treatment against bacterial soft
rot cause by Pectobacterium carotovorum.

Lebeouf-Little (2013) stated that, chemical control of

this soft rot agent are synthetic, such as Borax, thus
causes environmental and health hazard.
 Southernwood (Artemisia abrotanum) plant

Antimicrobial Property It has Coumarin- anti-repellant

• Terpenes 1.8-cineol property (Narayanaswamy, 2014).
• Davanone
• Eudesmanolides •Tannins
• Guianolides •Alkaloid
(Zafra-Polo & Blázquez, •Flavonoids
1991) •Phenolic acid
(Brodin, Alahyar, Hedner,
Sterner & Faergemann,
2007).
 THE ANTIBACTERIAL ACTIVITY OF
SOUTHERNWOOD (Artemisia abrotanum)
ETHANOLIC LEAF EXTRACT AGAINST
SOFT ROT AGENT (Pectobacterium
carotovorum)
Proponents

Poniones, Febeirlene C. Breguera, Arjay M.

Ordaneza, Ichiro M. Cabaya, Alaizza Cristine P.

Advincula, Zyndale Shelly V. Pepito, Rexielyn Mae L.

Agustin, Jahziel Kate C Segura, Jezza

Aquino, Simon Ray R.

Arceo, Sharmaine B.
 Conceptual Framework
Independent Variable Dependent Variable
Southernwood Ethanolic
Leaf Extract
a. 0.10 mg/mL (1.5 mL
extract & 1.5 mL
distilled water) Minimum Inhibitory
b. 0.15 mg/mL (2.25 mL Concentration
extract & 0.75 mL (MIC)
distilled water) Minimum Bactericidal
c. 0.20 mg/mL (3 mL of Concentration
pure extract) (MBC)
Positive Control
• Tetracycline 3 mg/mL
and 0.30 mg/mL
Negative Control
• Distilled water
 Statement of the Problem

1. What phytochemical components are present in the

Southernwood Ethanolic Leaf Extract used in this study?

2. What is the minimum inhibitory concentration and minimum

bactericidal concentration of Pectobacterium carotovorum
when treated with
a.) 0.10 mg/mL (1.5 mL extract added with 1.5 mL
distilled water)
b.) 0.15 mg/mL (2.25 mL extract added with 0.75 mL
distilled water) and
c.) 0.20 mg/mL (3 mL pure extract) of southernwood
ethanolic leaf extract?
Hypothesis

There is no significant difference on the Minimum

Inhibitory Concentration and Minimum Bactericidal

concentration of Pectobacterium carotovorum when treated

with Artemisia abrotanum and Pectobacterium carotovorum

when treated with Tetracycline.

Methods
 Transmittal letters
(Appendix A)

Collection of Southernwood Procurement and verification

leaves of Pectobacterium
(Appendix B) carotovorum
(Appendix B & C)
Verification of Southernwood
leaves
(Appendix C)

Southernwood ethanolic
leaf extraction and
Iodoform Test
(Appendix D)
 Phytochemical testing and
preparation of the concentrations
(Appendix E & Appendix F)

Agar making, subculture and incubation

of Pectobacterium carotovorum
(Appendix F)

Antibiotic Standard Preparation and

Application of treatment
(Appendix F)
 Data Collection

Analysis and interpretation of data

Decontamination and
Disposal
(Appendix H)

Final Report
Results
Minimum Inhibitory Concentration

0.10 mg/mL
0.15
0.20 mg/mLconcentration
concentration

Negative control Positive control:

3 mg/mL and 0.30 mg/mL
 Negative control Positive control:
3 mg/mL and 0.30 mg/mL

0.10 mg/mL 0.15 mg/mL 0.20 mg/mL

 0.10 mg/mL
0.15
0.20 mg/mL

Results
Minimum Bactericidal Concentration

Negative
Positive
control
Control:
0.30 mg/mL (300 µg/mL) and 3 mg/mL
Negative Control Positive Control
0.10 mg/mL
0.15 mg/mL

0.20 mg/mL
Discussions

 Phytochemical results:
 Alkaloids and Tannins

 Experimentation Results:
 MIC: 0.10 mg/mL
 MBC: 0.15 mg/mL

 Conformation:
 MIC and MBC results showed relation to the studies conducted by
Ijah and Oyebanji (2003), Karou et al. (2005), and Watson and
Preedy (2008)
Summary

 Phytochemical result: tannins and

alkaloids
 Minimum Inhibitory Concentration: 0.10
mg/ml
 Minimum Bactericidal Concentration: 0.15
mg/ml
Conclusion

In the light of the findings of the study, it is concluded that

the Southernwood ethanolic leaf extract has bactericidal effect

against Pectobacterium carotovorum and is a potential

treatment against the pathogen. It can be used as an alternative

for tetracycline, and other synthetic drugs of the same action, for

the treatment of P. carotovorum infection.

Recommendations

1. Perform a complete phytochemical determination.

2. Determine the specific chemical components and
quantity of each component by means of High
Performance Liquid Chromatography testing.
3. Conduct studies using Southernwood ethanolic
leaf extract against a wider range of
microorganisms with representations for bacteria,
fungi, and parasites that is of significance to
human health.
4. Determine the toxicity level of the extract.
References:
 Aguinaldo, A. M. (2005). Plant Screening, Phytochemical and Biological.

 Bais, H., & Sherrier, J. (2016). Plant Microbe Interactions. Google Books. Retrieved
27 February 2016, from https://goo.gl/N3ITR7

 Brodin, K., Alahyar, H., Hedner, T., Sterner, O., & Faergemann, J. (2007). In Vitro
Activity of Artemisia abrotanum Extracts Against Malassezia Spp., Candida albicans
and Staphylococcus aureus. Acta Derm Venereol Acta Dermato-Venereologica,
87(6), 540-542. Retrieved March 11, 2016 from http://goo.gl/9UFPQ0

 Charkowski, A., Condemine G., Expert D., Hayes C., Toth I., et al., (2012). The role
of secretion systems and small molecules in soft-rot Enterobacteriaceae
pathogenicity. Annu Rev Phytopathol. 2012;50:425-49. doi: 10.1146/annurev-phyto-
081211-173013. Retrieved January 21, 2016 from
http://www.ncbi.nlm.nih.gov/pubmed/22702350
 Elphinestone, John G., (2010) "Soft Rot and Blackleg of Potato." Technical
Informational Bulletin 21 Aug 1987: 1–8. Web.. Retrieved March 29, 2016
from http://www.cipotato.org/library/pdfdocs/TIBen15894.pdf

 Narayanaswamy, V. K., Gleiser, R. M., Kasumbwe, K., Aldhubiab, B. E.,

Attimarad, M. V., & Odhav, B. (2014). Evaluation of Halogenated Coumarins
for Antimosquito Properties. The Scientific World Journal, 2014, 1-6.
Retrieved February 26, 2016 from
http://www.hindawi.com/journals/tswj/2014/189824/

 Nezhad, M. H., Alamshahi, L., & Panjehkeh, N. (2012, October 2). Biocontrol
Efficiency of Medicinal Plants Against Pectobacterium Carotovorum,
Ralstonia Solanacearum and Escherichia Coli. Retrieved February 10, 2016,
from http://benthamopen.com/contents/pdf/TOPROCJ/TOPROCJ-3-3-46.pdf
 Sturluson, T. (n.d.). Southernwood Benefits, Uses and Side Effects. Retrieved
February 10, 2016, from http://www.herbal-supplement-resource.com/southernwood-
benefits.html

 Salunkhe, D. K., & Kadam, S. S. (2005). Handbook of vegetable science and

technology: Production, composition, storage, and processing (Vol. 86). New York:
Marcel Dekker. Retrieved April 2, 2016 from https://goo.gl/fyg2Iq

 Simeon, A., & Abubakar, A. (2014). Evaluation of Some Plant Extracts for the Control
of Bacterial Soft Rot of Tubers. AJEA American Journal of Experimental Agriculture,
4(12), 1869-1876. Retrieved April 2, 2016 from https://goo.gl/QmdYsB

 Zafra-Polo, M. C., & Blázquez, M. A. (1991). Antiinflammatory activity of

sesquiterpene lactones from Artemisia barrelieri in rats. Phytother. Res. Phytotherapy
Research, 5(2), 91-93. Retrieved January 24, 2016 from http://goo.gl/TpGyO
Thank you!
Computations
 
Total amount of powdered Southernwood leaves applied with inclusion-exclusion criteria
= 710 grams, soaked in ethanol with 1:5 weight-to-volume ratio 710 x 5= 3, 550 mL.

 
The extract was subjected to Rotary Evaporation test, which gave a total volume of 100
mL of solution.

The 20 mg/100 mL is simplified to 0.20 mg/mL which is the concentration of the pure
extract solution.
 
The pure stock solution was further diluted to test different concentrations
 
1.5 mL extract diluted with 1.5 mL distilled water:
 

 
, added with 1.5 mL distilled water;
 

 
2.25 mL extract diluted with 0.75 mL distilled water;
 

 
, added with 1.5 mL distilled water;
 

 
3 mL of pure stock solution (0.20 mg/mL or 1:5)