Anti-Microbial Assay of on Selected Manure and Stagnant Water REGIONAL LEVEL (1)

8/9/2019 Anti-Microbial Assay of on Selected Manure and Stagnant Water REGIONAL LEVEL (1)

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IV. RESEARCH PLAN

A. Statement of the Problem

This study will determine the antimicrobial assay of the combined Tinospora rumphii

Boerl (Makabuhay stem), Psidium guajava Linn.(Guava leaves), and Citrus microcarpa

(Calamansi) upon application on selected manure and stagnant water

!pecifically, it will seek to answer the following "uestions#

1) $hat is the effect of the combined Tinospora rumphii Boerl (Makabuhay stem), Psidium

guajava Linn. (Guava leaves), and Citrus microcarpa(Calamansi) on its application on selected

manure and stagnant water in terms of#

11) odor before and after application% and

1&) 'one of inhibition of bacteria

&) $hich among the three Treatments is the most effective in terms of#

&1) odor before and after application% and

&&) 'one of inhibition of bacteria

) *s there a significant difference between the three setups in terms of#

1) odor before and after application

B. Statement of Hypotheses

1) The combined Tinospora rumphii Boerl (Makabuhay stem), Psidium guajava Linn.(Guava

leaves), and Citrus microcarpa (Calamansi) has no effect on its application on selected

manure and stagnant water in terms of#



&) The three setups are not effective in terms of#


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) There is no significant difference between the three setups in terms of#

&1) odor before and after application

C. Methodology

The materials will be used in study are the following# one hundred grams of makabuhay

(Tinospora rumphii Boerl) stem, one hundred grams of guava(Psidium guajava Linn.) leaves,

one hundred grams of calamansi (Citrus microcarpa), water, five beakers, one sterili'ed bottle,

weighing scale, mortar and pestle, gau'e, syringe, a stirring rod, three filter papers, three slides,

normal saline solution (+!!), gloves, masks, microscope, four glass container, casserole and

stove

eneral Pro!ed"re

The calamansi (Citrus microcarpa), guava (Psidium guajava Linn.) leaves and

makabuhay (Tinospora rumphii Boerl) stem will be collected in the researchers back yard -ne

hundred grams of guava leaves and one hundred grams of makabuhay stem will be pounded

using mortar and pestle The solution will be prepared using the decoction process The decocted

guava leaves and makabuhay stem will be filtered by using a filter paper .ifty m/ of decocted

guava leaves and makabuhay stem will be kept in a beaker -ne hundred grams of calamansi will

be e0tracted using gau'e and fifty m/ of calamansi e0tract will be collected in a beaker The

three solutions will be combined in a larger beaker and will be stirred in a clockwise direction

fterwards, the mi0ture will be transferred in a sterili'ed bottle n actual test will be used to

obtain the results in terms of its odor by applying the three treatments in the piggeries, poultries


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and stagnant water Three samples from each area will be collected for the analysis of the

bacteria n electronic microscope will be used in observing the bacteria present in the samples

+ormal !aline !olution (+!!) will be needed in observing the specified materials under the

electronic microscope

#est$ng Pro!ed"re

The following setups will be# 2efore application !etup 1 (+egative control), fter

pplication% !etup & (3ositive Control) and !etup (40perimental Control)

*n gathering data, an actual test will be conducted on the piggeries, poultries and stagnant

areas in order to test the odor before and after the application These will be rated by thirty

human respondents

Three samples will be collected including pig manure, chicken manure and stagnant

water and will be placed in a clean container wet mount analysis will be used in testing the

anti5microbial activity of the treatments Three samples will be prepared using slides with an

+!! small amount of waste material will be placed in a slide Then this will be observed

under the microscope +e0t, small amount of the treatments will be poured on the specimen

4ach of the treatments will be tested and analy'ed in the laboratory to observe the presence of

the bacteria in the samples

R$s% and Safety


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Gloves will be used while performing the e0periment Masks will be also used before

rating the odor of the selected manure and stagnant water in order to avoid disease5causing

bacteria that could be taken from the areas

&ata Analys$s

The statistical tool that will be used in the study is 7andomi'ed 2locks +-8

&. B$bl$ography

'ebs$te

Cabral, 93! (&:1:) $ater Microbiology 2acterial 3athogens and $ater International Journal

of Environmental esearch and Pu!lic "ealth, ;, :?i@erph;1:>=) Molasses.#rolier Enc$clopedia of %no&ledge(p1


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Foesnt curiosity bother you about the foul odor when you pass by a slaughter house

Today, the most common problem in slaughter houses or pig farms is the increasing rate of

undesirable odor and diseases caused by rotting of organic matter of microorganisms This

situation is provoked when the wastes are being pumped to the canals and other stagnant areas

which cause diseases to humans and animals

Most of the farmers in rural areas have meager income Money cannot be easily earn in

the farm because of some problems encountered Most chickens and pigs tend to lay their

droppings everywhere and anywhere 2ecause of this, it gave way for pathogenic

microorganisms to thrive and multiply around the areas which caused diseases following an

undesirable odor to the pigs and poultry farms *t also results to pollution of water supplies which

is a ma@or health problem today +ot only may sanitary sewage be discharge into water, but

comple0 wastes from agriculture may be also *n increasing volume such substances as animal

and vegetable matter, are released from poultry, processing plants, canals, oil fields and farms

2acterial diseases also affect animals, and they are responsible for various diseases of crop

plants

The present condition in slaughter houses, poultry farm and stagnant water stress the need

for a fast and accurate solution for the problem Thus, this study aimed to develop an effective

anti5microbial treatment from combined Tinospora rumphii Boerl (Makabuhay), Psidium

guajava Linn. (Guava), and Citrus microcarpa(Calamansi)

B. eneral Spe!$f$! (b*e!t$+e

The specific ob@ectives of this study were the following#


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:), +treptococcus

salivarius (MTCC 1>), Lacto!acillus acidophilus (MTCC 66;), +treptococcus sanguinis

(TCC 1:==


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bacterial pathogens Ma0imum antibacterial activitywas observed against +. sanguinis(& mm)

and lowest activity against +. salivarius (1; mm) The phytochemical analysis revealed the

presence of alkaloids, flavonoids, tannins, phenols, saponins, glycosides/ amino acids and

steroids which an accountable for its antimicrobial potential The results validate the traditional

uses of T. cordifoliain treatment of dental diseases

This stud$ &as related to present stud$ !ecause it focuses on the anti!acterial activit$ of

Tinospora cordifolia (/a1a!uha$) e0tract against dental pathogens. This stud$ concentrate on

T. cordifolia that possess al1aloids, flavonoids, tannins, phenols, saponins, gl$cosides, amino

acidsand steroids &hich are 1no&n for its antimicro!ial potential.

*n the study of Mahalel (&:1&) entitled Kntibacterial !ensitivity for some Chemically

Fiverse !teroidal GlycosidesIn 2itro3, susceptibilities (in vitro) of the steroidal glycosides were

tested againstBacillus cereus, +taph$lococcus aureus, +erratia marcescens,Escherichia coli and

Pseudomonas putida using the disc diffusion method These steroidal glycosides were identified

as alpinoside C (1) Lkid@olanin5455F5glucopyranosyl5(1N6)55F5glucopyranosyl5(1N6)55F5

oleandropyranosyl5(1N6)55F5cymaropyranosyl5(1N6)55F5cymaro5pyranosideO% Ghalakoside

(&) Lcardiac glycosideO and pentandroside () L(&&!,&=!)51


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1=

This stud$ &as related to present stud$ !ecause +teroidal #l$cosides &as proven to have

remar1a!le suscepti!ilities against #ram negative and #ram positive.

Fisinfectant properties of essential oils from +alvia officinalis/ cultivated in Tunisia

were evaluated 4ssential oils were obtained by hydro5distillation of the aerial part of +alvia

officinalis / cultivated in !fa0 gardens, Tunisia The obtained oils were analy'ed by gas

chromatographyHmass spectrometry (GCHM!) and 66 compounds were identified !trong

bactericidal and fungicidal effects were shown using the +CC/! broth dilution, 4+ 1&;= and

4+ 1&;< stOandard methods The minimal cidal concentrations (MCCs) values ranged from

::1 to :&= Q/ m/

R1

The essential oils concentrations of :=J and 1J (v?v) resulted in a

reduction in viability higher than = and 6 log units per m/ for the standard bacteria and fungi,

respectively, within a contact time of = min Asing an air sampler and an aroma dispenser,

vaporisation of :&= m/ mRof +. officinalisessential oils resulted in (;&J, ;J and ;:J) and

(=6J, ==J and ==J) reduction of the total microbial count and the total count of yeasts and

moulds, after a residence time of 1 h, < h and &6 h in a selected testing room, respectively +.

officinalisessential oils showed a potent vapour activity against a panel of bacteria, yeasts and

fungi This supported their use as a natural eco5friendly disinfectant to manage airborne microbes

(2oua'i', &::>)

This stud$ &as related to the present stud$ !ecause the essential oils &ere proven as an

effective disinfectant against a panel of !acteria, $east and fungi.

*n the study of Su (&::=) entitled Kntimicrobial activity of berberine alone and in

combination with ampicillin or o0acillin against methicillin5resistant !taphylococcus aureus,

the antimicrobial activity of berberine was investigated The main antibacterial substance of

Coptidis rhi'oma (Coptis chinensis .ranch) and 3hellodendri corte0 (3hellodendron amurense

7uprecht), against clinical isolates of M7!, and the effects of berberine on the adhesion to


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1:J

verage :J 1::J =J
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0

0.2

0.4

0.6

0.8

1

1.2

Before Application

Setup 1

Setup 2

The results of the present study was supported by Cowan (&:::) which stated that guava

(Psidium guajava Linn.) posses alkaloid and flavonoids which are phytochemical compounds

,$g"re 4. >one of Inh$b$t$on of ba!ter$a before and after appl$!at$on


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&>

that have been found in vitro to be an effective anti5microbial treatment against a wide array of

microorganisms

Table < below shows the bacteria found in the selected manure and stagnant water after

the tests conducted

#able 7. Ba!ter$a ,o"nd $n the sele!ted man"re and stagnant ater

Lo!at$on Ba!ter$a

1 3ig Manure Cr$ptosporidium, Clostridium Perfringens,

Escherichia coli (ecal Coliform), .ecal streptococci

& Chicken Manure Clostridium Perfringens, Escherichia coli (ecal

Coliform), .ecal streptococci, Cr$ptosporidium

!tagnant $aterCr$ptosporidium, Escherichia coli (.ecal coliform),

Enterococci

VIII. C(NCL)SI(N

2ased on the foregoing results and analysis of data, this study found out that the

combined Tinospora rumphii Boerl (Makabuhay), Psidium guajava Linn.(Guava), and Citrus

microcarpa(Calamansi) had a positive effect on its application on selected manure and stagnant

water in terms of 1) -dor before and after application% and &) Vone of *nhibition of bacteria

The combined Tinospora rumphii Boerl (Makabuhay), Psidium guajava Linn.(Guava),

and Citrus microcarpa(Calamansi) was found to be the most effective treatment in terms of odor

of test samples before and after application The 3ositive Control (Fome0) is the most effective

in terms of 'one of inhibition of bacteria

*n addition, there is a significant difference between the three setups in terms of odor

before and after application


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:

I?. REC(MMEN&A#I(NS

$ith the results and conclusion presented, this study forwards the following

recommendations for further studies .irst, it is highly recommended to use the combined e0tract

of makabuhay stem, guava leaves and calamansi as disinfectant on areas with foul odor like pig

pens, poultry farms and stagnant water .urther studies can also be made to test the effectiveness

of makabuhay stem, guava leaves and calamansi as an anti5microbial treatment dditional

studies should also be conducted to determine other methods of applying the combined e0tract of

makabuhay stem, guava leaves and calamansi $hen a wet mount analysis was used for the test,

gross e0amination should be used in interpreting the data being gathered after the test

?. BIBLI(RAPH1

Boo%s

9oa"uin, C, /agun'ad, C, 7abago, / (&::) unctional Biolog$ Due'on City# 8ibal

3ublishing Eouse, *nc

Burian, 9C (&:1:)ma*ing "ealing Plants Manila# 3hilippine 3ublishing Eouse

En!y!loped$a

Grolier *nternational (1>>=) #rolier Enc$clopedia of %no&ledge(p1


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1

'ebs$tes

nwar et al (&::) Soung leaf e0tract of the plant possesses antibacterial and antifungal

properties, 7etrieved ugust >, &:1, from http#??wwwmpbdinfo?plants?psidium5

gua@avaphp

rcha 8, +avneet Gautam, ! (&:1) !creening of ntibacterial ctivity ofTinospora

cordifoliaMiers 40tracts gainst Fental 3athogens/7etrieved ugust 1, &:1, from

http#??scialertnet?fullte0t?doi W@pt&:1&6

9ackson, + (&:1) Essential 4il 4dor emoval. 7etrieved ugust 1, &:1 from

http#??wwwehowcom?infoX


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Mahalel, U. (2012). ntibacterial !ensitivity for some Chemically Fiverse !teroidal

Glycosides in 2itro.Journal of griculture 5 +ocial +ciences, &=5&

Su, E et al (&::=) ntimicrobial activity of berberine alone and in combination with ampicillin

or o0acillin against methicillin5resistant !taphylococcus aureus Journal of /edicinal

ood, , 6=65

Documents

Anti-Microbial Assay of on Selected Manure and Stagnant Water REGIONAL LEVEL (1)