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Republic of the Philippines Sultan Kudarat Polytechnic State College Science Laboratory High School ACCESS, EJC Montilla,

acurong City

his study

Antimicrobial Activity of Birds Nest Fern Extract Against Escherichia Coli


is presented to !i"ision Science #air $%%&' RESEARCHERS:

(reylle (epitel Earl Ada) Prado Rona Angeli*ue Manuel

RESEARCHER A !"SER:

Prof' Merlyn Leysa


Septe)ber $%%&

!edication his hu)ble +or, is dedicated to all -ho ha"e co))itted the)sel"es o the i)pro"e)ent .f the life of our people'

AC#N$%&E 'E(EN)
he researchers recogni/e +ith the gratitude and appreciation their indebtness to the follo+ing persons and institutions +ho ha"e )ade "aluable contribution to the successful co)pletion of this study0 o the Prof' Merlyn Leysa, our researcher ad"iser for her ti)e, for the guidance, patience in chec,ing drafts, encourage)ent and for gi"ing enough consideration1 o the SKPSC faculty and staff and !r' Her)inia !u)adag for the untiring support e2tended to us1 o the SKPSC Research Center , +ho pro"ided us the )aterials and e*uip)ent used in the conduct of this study1 o our class)ates and friends +ho pro"ided fro) the co)panionship and inspiration for the co)pletion of this study1 o our belo"ed parents +ho are supporting us for helping and pro"iding all +e need and1 Most of all, 3od, +ho has endo+ed the researchers +ith intellectual gifts and strong faith +hich sustained the) through the different stages of their research +or,'

)he Researchers

ABS)RAC)

(reylle (epitel, Earl Ada) Prado, and Rona Angeli*ue Manuel' A pro4ect design *Antimicrobial Activity of Birds Nest Fern Extract Against Escherichia Coli+' Sultan Kudarat Polytechnic State College5 Laboratory High School, Research Center , EJC Montilla, acurong City ' Septe)ber $%%&' Advisers0 Prof' Merlyn Leysa' he study Anti)icrobial Acti"ity of (ird6s 7est #erns E2tract against Escherichia Coli8 generally ai)ed to deter)ine the potential of bird6s nest fern as an antibiotic for Escherichia Coli' Specifically the study ai)ed to0 9' !eter)ine the sensiti"ity of E' coli to bird6s nest fern e2tract through Kirby5(auer antibiotic testing' $' !eter)ine the treat)ent +here the speci)ens of bacteria are )ost sensiti"e to :' !eter)ine the significant differences a)ong treat)ents and replications conducted' he )a4or findings of this study +ere as follo+s0 9' reat)ent 9;pure birds nest e2tract< got a general )ean of its => +ith ?&'@? )) +hile reat)ent $ ;? )l bird6s nest e2tract A ?)l of sterili/ed +ater< got a general )ean of %'BC))' he control cefuro2i)e got a general )ean of ?9'99))' $' Escherichia coli sho+ed high sensiti"ity +hen e2posed to pure bird6s nest fern e2tract' :' >t is therefore found out that the treat)ent 9 is +here Escherichia coli is )ost sensiti"e to' @' (ased on the Analysis of Dariance, treat)ents ha"e significant differences a)ong each other in all trials conducted, +hile the replicates and trials are not significant' After the $@5hour incubation, the dia)eter of the /one of inhibition of E' coli +ere deter)ined as positi"e or negati"e to be sensiti"e to bird6s nest fern' (ased on the findings of the test conducted, bacteria E' coli is fa"orably sensiti"e to treat)ent 9, +hich is co)posed of pure bird6s nest e2tract fro) its lea"es as it +ere re)ar,ed sensiti"e' he treat)ents sho+ed highly significant differences +hile the replicates and trials are "ery precise and sho+ed not )uch significance' Li,e+ise, the co)parison bet+een the control group and the treat)ents used cannot be directly )easured since the control ;cefuro2i)e< is )anufactured considering the dosage for hu)an consu)ption +hile bird6s nest fern e2tract is )ade to co)bat E' coli in direct contact +ith the bacteria itself' 7e"ertheless, the study generally pro"ed the great potential of (ird6s 7est fern e2tract to be the radical antidote for the threatening public health night)ares, the E' coli'

)AB&E $F C$N)EN)S > LE PA3E EEEEEEEEEEEEEEEEEEEEEEEEEEEE'i !edication EEEEEEEEEEEEEEEEEEEEEEEEEEEEE''ii Ac,no+ledge)entEEEEEEEEEEEEEEEEEEEEEEEEEE''iii Abstract EEEEEEEEEEEEEEEEEEEEEEEEEEEEE'''''i" HE PR.(LEM EEEEEEEEEEEEEEEEEEEEEEEEEE''9 (ac,ground of the Study EEEEEEEEEEEEEEEEEEEEEEE''9 .b4ecti"es EEEEEEEEEEEEEEEEEEEEEEEEEEEE''''$ Significance of the Study EEEEEEEEEEEEEEEEEEEEEEE''': Hypotheses EEEEEEEEEEEEEEEEEEEEEEEEEEEE''': Scope and !eli)itations EEEEEEEEEEEEEEEEEEEEEEE'''': !efinition of Key ter)s ''EEEEEEEEEEEEEEEEEEEEEEE'''@ RED>E- .# RELA E! L> ERA FRE A7! S F!>ES EEEEEEE'''''''''''?

RESEARCH ME H.!.L.3GEEEEEEEEEEEEEEEEEEE' E'9$ Research !esign EEEEEEEEEEEEEEEEEEEEEEEEE''''9$ Material 3athering ProcedureE'' EEEEEEEEEEEEEEEEEE''''''''''''9$ Preparation .f E2peri)ent Set5FpE'E'EEEEEEEEEEEEEEE'''EE9$ Materials and E*uip)ent E'EEEEEEEEEEEEEEEEEEEEEE'''9: Methodology '' EEEEEEEEEEEEEEEEEEEEEEEEEEE''9@ !ata Collection Procedure EEEEEEEEEEEEEEEEEEEEEE''9? Statistical Analysis EEEEEEEEEEEEEEEEEEEEEEEEE''''''9? RESFL S A7! !>SCFSS>.7SEE'EEEEEEEEEEEEEEEEEE'9B SFMMARG .# #>7!>73S, C.7CLFS>.7, A7! REC.MME7!A >.7EEE$@ #indings EEEEEEEEEEEEEEEEEEEEEEEEEEEEEE'$@ Conclusions EEEEEEEEEEEEEEEEEEEEEEEEEEE'''''''$? Reco))endationsEEEEEEEEEEEEEEEEEEEEEEEEEE'''$? APPE7!>CES L> ERA FRE C> E!

CHA,)ER )he ,roblem


Moderni/ation li,e ad"ance)ent in )any fields gro+ faster, definitely people are a+are of this' >t leads to open their )inds to disco"er and e2pand their ,no+ledge through de"eloping their concepts to sol"e different health proble)s' oday, antibiotic resistance is one of the )a4or proble)s of )edical practioners' Antibiotic resistance is the ability of a )icroorganis) to +ithstand the effects of antibiotics' >t is a specific type of drug resistance' Antibiotic resistance e"ol"es "ia natural selection acting upon rando) )utation, but it can also be engineered by applying an e"olutionary stress on a population' .nce such a gene is generated, bacteria can then transfer the genetic infor)ation in a hori/ontal fashion ;bet+een indi"iduals< by plas)id e2change' >f a bacteriu) carries se"eral resistance genes, it is called )ultiresistant or, infor)ally, a superbug' he ter) anti)icrobial resistance is so)eti)es used to e2plicitly enco)pass organis)s other than bacteria' Antibiotic resistance can also be introduced artificially into a )icroorganis) through transfor)ation protocols' his can aid in i)planting artificial genes into the )icroorganis)' >f the resistance gene is lin,ed +ith the gene to be i)planted, the antibiotic can be used to ,ill off organis)s that lac, the ne+ gene' he researchers ha"e in"estigated the anti5bacterial properties of "arious plants atte)ting to de"elop a ne+ antibiotic out of natural substances that could be used by co))unities' (irds nest fern is effecti"e against infecting Escherichia Coli and Pseudo)onas aeruginosa' hey content the potent antibiotic co)ponents li,e al,aloids, fla"anoids, saponins and tannins' hey ha"e strong defensi"e )echanis)' he study atte)pts to deter)ine the potential of (ird6s nest fern e2tract by e"aluating the sensiti"ity of the Escherichia coli sub4ected to Kirby5(auer Antibiotic esting'

$b.ectives of the st/dy 'eneral ob.ectives


he study generally ai)ed to deter)ine the potential of bird6s nest fern e2tract as an antibiotic for Escherichia Coli.

S0ecific ob.ectives
Specifically the study ai)ed to0 9 !eter)ine the sensiti"ity of E' coli to bird6s nest fern e2tract through Kirby5(auer antibiotic testing

$' !eter)ine the treat)ent +here the speci)ens of bacteria are )ost sensiti"e to :' !eter)ine the significant differences a)ong treat)ents and replications conducted' Significance of the St/dy Specialists esti)ate that at least &%H of antibiotic prescriptions are unnecessary or inappropriate' Mutation of bacteria happens and occurs anyti)e by the transfer of resistance genes fro) the other bacteria' Perhaps a ne+ re"olution of antibiotic is declared to be a necessity' he study IAnti)icrobial Acti"ity of (ird6s 7est #erns E2tract against Escherichia Coli has a "ital i)portance in deter)ining the effecti"e and cheap antibiotic out of co))only found in the locality li,e bird6s nest fern +hich can also eradicate different ,inds of bacteria, according to past references' >t is also significance in a"oiding the use of har)ful che)icals +hich can pollute the air, ,ill other organis) than the target pest and can destroy the biodi"ersity' Hy0otheses H1: (ird6s 7est #erns E2tract has no antibacterial effect on Escherichia Coli, H-: (ird6s 7est #erns E2tract has antibacterial effect on Escherichia Coli, Sco0e and limitations of the St/dy his study +as li)ited only on the detection of the sensiti"ity of Escherichia Coli to (ird6s 7est #ern e2tract and to the stated ob4ecti"es of the research study' his study +as conducted at Sultan Kudarat Polytechnic State College5 Laboratory High School Research Center , EJC Montilla, acurong City fro) Septe)ber 9$59B $%%&'

efinitions of #ey )erms


Agar2 is a gelatinous substance chiefly used as culture )ediu) for )icrobiological +or,' Antibiotic5 used to ,ill bacteria that gi"e different diseases' Antimicrobial agent5 the bacteria causing agent' >ts the transfer of )ediu) to )ediu)' Birds nest fern2 is a tropical and sub5tropical plant +hich is nati"e to the rain forests of Asia, Africa, >ndia , Polynesia and Australia ' Cef/roxime2 is a second5generation cephalosporin antibiotic that has been +idely a"ailable' Escherichia coli5 can cause serious food poisoning in hu)ans, and are occasionally responsible for costly product recalls'

Extract2 co)es fro) the lea"es or any parts of a plant that +ere sub4ected to pounding and e2tracting' "nc/bation 2 he de"elop)ent of an infection fro) the ti)e the pathogen enters the body until signs or sy)pto)s first appear' #irby2Ba/er antibiotic testing 3#B testing or dis4 diff/sion antibiotic sensitivity testing52 is a test +hich uses antibiotic5i)pregnated +afers to test +hether particular bacteria are susceptible to specific antibiotics' Sterili6ation 7 a process used to ensure that the )aterials are safe and clean' 8one of inhibition2 his is an area around a paper dis, or colony of bacteria or )old +here no other organis)s are gro+ing'

RE!"E% $F RE&A)E

CHA,)ER ""

&")ERA)9RE AN

S)9 "ES

Related &iterat/re
Escherichia coli is a 3ra) negati"e bacteriu) that is co))only found in the lo+er intestine of +ar)5blooded organis)s ;endother)s<' Most E' coli strains are har)less, but so)e, such as serotype .9?C0HC, can cause serious food poisoning in hu)ans, and are occasionally responsible for costly product recalls' he har)less strains are part of the nor)al flora of the gut, and can benefit their hosts by producing "ita)in K$, or by pre"enting the establish)ent of pathogenic bacteria +ithin the intestine' E' coli are not al+ays confined to the intestine, and their ability to sur"i"e for brief periods outside the body )a,es the) an ideal indicator organis) to test en"iron)ental sa)ples for fecal conta)ination' he bacteria can also be gro+n easily and its genetics are co)parati"ely si)ple and easily5)anipulated or duplicated through a process of )etagenics, )a,ing it one of the best5studied pro,aryotic )odel organis)s, and an i)portant species in biotechnology and )icrobiology' A strain of E' coli is a sub5group +ithin the species that has uni*ue characteristics that distinguish it fro) other E' coli strains' hese differences are often detectable only on the )olecular le"el1 ho+e"er, they )ay result in changes to the physiology or lifecycle of the bacteriu)' #or e2a)ple, a strain )ay gain pathogenic capacity, the ability to use a uni*ue carbon source, the ability to inhabit a particular ecological niche or the ability to resist anti)icrobial agents' !ifferent strains of E' coli are often host5specific, )a,ing it possible to deter)ine the source of fecal conta)ination in en"iron)ental sa)ples'

#or e2a)ple, ,no+ing +hich E' coli strains are present in a +ater sa)ple allo+s )a,ing assu)ptions about +hether the conta)ination originated fro) a hu)an, another )a))al or a bird' 7e+ strains of E' coli e"ol"e through the natural biological process of )utation, and so)e strains de"elop traits that can be har)ful to a host ani)al' Although "irulent strains typically cause no )ore than a bout of diarrhea in healthy adult hu)ans, particularly "irulent strains, such as .9?C0HC or .9990(@, can cause serious illness or death in the elderly, the "ery young or the i))unoco)pro)ised' Biology and biochemistry Escherichia coli cells propel the)sel"es +ith flagella ;long, thin structures< arranged as bundles that rotate counter5cloc,+ise, generating tor*ue to rotate the bacteriu) cloc,+ise'E' Coli is 3ra)5negati"e, facultati"e anaerobic and non5sporulating' Cells are typically rod5shaped and are about $ )icro)etres ;J)< long and %'? J) in dia)eter, +ith a cell "olu)e of %'B 5 %'C J):' >t can li"e on a +ide "ariety of substrates' E' coli uses )i2ed5acid fer)entation in anaerobic conditions, producing lactate, succinate, ethanol, acetate and carbon dio2ide' Since )any path+ays in )i2ed5acid fer)entation produce hydrogen gas, these path+ays re*uire the le"els of hydrogen to be lo+, as is the case +hen E' coli li"es together +ith hydrogen5 consu)ing organis)s such as )ethanogens or sulfate5reducing bacteria' .pti)al gro+th of E' coli occurs at :CKC but so)e laboratory strains can )ultiply at te)peratures of up to @&KC' 3ro+th can be dri"en by aerobic or anaerobic respiration, using a large "ariety of redo2 pairs, including the o2idation of pyru"ic acid, for)ic acid, hydrogen and a)ino acids, and the reduction of substrates such as o2ygen, nitrate, di)ethyl sulfo2ide and tri)ethyla)ine 75o2ide' he Bird:s Nest Fern, Asplenium nidus, is a tropical and sub5tropical plant +hich is nati"e to the rain forests of Asia, Africa, >ndia , Polynesia and Australia ' >n the +ild, these largest of epiphytes li"e up in the cro+ns of trees' heir )eans of nourish)ent is fro) nutrients found in dust particles and rain +hich is collected in the plants deep, funnel5shaped rosette of lea"es' he genus na)e Aspleniu) ;Spleen+ort< is deri"ed fro) the 3ree, +ords LaL )eaning LnotL, and LsplenL, )eaning LspleenL and refers to the )edicinal properties the plant is supposed to ha"e' he specific epithet or species na)e LnidusL )eans La nestL, hence the co))on na)e (irdLs 7est #ern' he (irdLs 7est #ern belongs to the LAspleniaceaeL fa)ily under the fern order LPolypodialesL' he leathery, tongue5li,e, glossy, pale green, +a"y5edged fronds of this fern are borne in a tight rosette at the rhi/o)e, for)ing a bo+l5shaped clu)p +ith a fibrous center, +hich to )any rese)bles a birdLs nest, or at least a see)ingly sheltered spot for a birdLs nest' Each frond is entire or undi"ided and has a pro)inent, polished rib running do+n the center of the frond' he ne+ fronds uncurl fro) the hairy, dar, scaled cro+n of the rosette

in typical fern )anner' hey are "ery soft and brittle +hen young and )ust not be handled' As the ne+ fronds gro+ in, the older ones gradually die' Mature fronds can gro+ to : feet in length and M inches in +idth, and under ideal conditions, a plant can reach @ to B feet in height, but, under nor)al ho)e conditions the plant rarely e2ceeds 9? inches in height' he ste)s of each frond are stu)py and erect' Another co))only gro+n "ariety is Aspleniu) crispafoliu), +hich has deeply ruffled fronds' ,hytochemical Content of &eaf Extract Al,aloids are naturally occurring che)ical co)pounds containing basic nitrogen ato)s' >t protects plant fro) herbi"ores' hey are anti5 spas)odic, anti5infla))atory,tran*uili/er and "asodilator' #la"onoids ;or biofla"onoids<, also collecti"ely ,no+n as Dita)in P and citrin, are a class of plant secondary )etabolites' hey are anti5allergic and anti)icrobial and anti5infla))atory' Saponins, group of naturally occurring oily glycosides that foa) freely +hen sha,en +ith +ater' hey occur in a +ide "ariety of plants, including acacia, soap+ort, soaproot, California pig+eed, and )any others' Saponins ha"e been, and so)eti)es still are, used as cleaning agents and as foa) producers, notably in fire5e2tinguishing fluids' hey ha"e a bitter taste annins , group of naturally occurring oily glycosides that foa) freely +hen sha,en +ith +ater' hey are anti)icrobial, protect against cancers and decrease blood lipids'

CHA,)ER """
Research (ethodology
his chapter discusses different )ediu)s utili/ed in the research design and studies as +ell the procedures and )ethods used in the conduct of the study' Also included are the )ethods in gathering data as +ell as the statistical tools in the research' Research esign he study is e2peri)ental and co)parati"e in nature' he study +as conducted using the Analysis of Dariance ;A7.DA< of Co)pletely Rando)i/ed !esign ;CR!< +ith the three treat)ents replicated three ti)es in three trials +ith the sa)e cultured bacteria per replication' he treat)ents +ere as follo+s0 reat)ent 9 5pure bird6s nest fern e2tract reat)ent $ 5? )l bird6s nest fern e2tract A ? )l +ater solution Control 5 Cefuro2i)e

(aterial 'athering ,roced/re he "ariables used in the study +ere bird6s nest fern and Escherichia coli' he bacteria +ere collected fro) the urine o"ersa+ by SKPSC Research Center for culturing reasons' (ird6s nest fern +ere collected at SKPSC garden and +ere gathered fresh for this contains )ore e2tract ,re0aration of Ex0eriment Set290 Escherichia Coli +ere cultured in the SKPSC Research Center for the study' he three treat)ents, replicated thrice, +ere placed rando)ly on e"ery Petri dish +ith the cultured bacteria in it' Application of e2tract +as ad)inistered by i)pregnating (ird6s 7est fern e2tract unto sterile paper dis,s' hree trials +ere conducted for the "alidity for the study'

(aterials and E;/i0ment he follo+ing +ere used for the conduct of the study0 For Extracting: !igital +eighing )achine Mortar and pestle Cheesecloth 3raduated cylinder Sterili/ed +ater (urner E)pty and clean bottle fern Crucible tongs For Sensitivity )esting: (unsen burner test tube >noculating loop incubator Sterile cotton s+ab +eighing scale Petri dish Erlen)eyer flas, #orceps hot plate

(id6s 7est

#oil paper Auto cla"e Sterile paper discs Mueller5Huinton agar ;$H< Sodiu) chloride etracycline i)pregnated filter5paper disc (acteria ;Escherichia Coli< Mc#arland Standard Agar /sed for inc/bating organisms: (lood agar MacCon,y Agar (ethodology: he follo+ing )ethods +ere used in the study0 Extraction of Birds Nest fern: 9' Collect the )aterials needed and prepare the setup' $' Pound the $% gra)s of bird6s nest lea"es fern using the )ortar and pestle :' S*uee/e the pounded lea"es using the cheesecloth to obtain the e2tract' @' reat)ent 9 re*uires concentrated a)ount of bird6s nest fern e2tract and re*uires no boiling' #or the treat)ent $, add ? )l solution of pure bird6s nest fern e2tract to ? )l sterili/ed +ater' !ispense the )i2ture unto a test tube and set to boiling +ater' ?' After obtaining the needed e2tract for both treat)ents, store solutions securely unto sterili/ed e)pty bottles and seal' #irby2Ba/er Antibiotic testing 3direct method5 9' 3ro+th for pri)ary plate5 inoculate into 7SS $' Strea, to susceptible plate' E'coli MH +ith $H 7aCL :' Let the inoculu) dry for :5? )inutes +ith the plates closed' @' Placed the sterile paper disc i)pregnated +ith respecti"e bird6s nest fern e2tracts and cefuro2i)e;control< on the agar plate +ith sterile forceps' Press do+n securely to ensure e"en contact' 5!isc )ust be e*uidistant +ith each other to allo+ /one of inhibition ;=>< in )illi)eters ?' >ncubate plates i))ediately or +ithin :% )inutes for $@ hours at degrees Celsius in an in"erted position' B' Read after $@5hour incubation' ata Collection ,roced/re

8one of "nhibition' After $@ hours, )easure the dia)eter, in )illi)eters, of the region +here the Escherichia Coli +ere inhibited after $@ hours of e2posure to the treat)ents' =one of inhibition +ill deter)ine the re)ar,s, +hether the bacteria +ere resistant or sensiti"e to the treat)ents' #or the control, it +as based on the antidote6s S>R legend ;on hospital standards<' Positi"e ;A< for => N sensiti"e 7egati"e ;5< for => N resistant Statistical Analysis of ata he /one of inhibition of each replicates and the )ean of the three replicates per treat)ents +ere co)puted' he data gathered +ere used in ans+ering the *uestion0 Which treatment will give the best result in terms of antibacterial effect on Escherichia Coli after 24 hours? !ata gathered in this study +as statistically analy/ed using the Analysis of Dariance of Co)pletely Rando)i/ed !esign' he test for significant differences a)ong treat)ents )eans is sub4ected to !uncanOs Multiple Range est ;!MR <

CHA,)ER "!
RES9&)S AN "SC9SS"$N his chapter presents the results of the tests conducted to deter)ine the potential of bird6s nest fern to be an antidote for Escherichia

coli and the statistical analysis perfor)ed on the data gathered' >t is also presents the interpretations of the results of the analysis' =one of inhibitions of Escherichia coli $@ hours after contact to bird6s nest fern e2tract for the different trials are being sho+n in the follo+ing table0 able 9' Results of the sensiti"ity testing for the first trial'
Treatments R1 R2 R3 Total Mean Remarks Zone of Inhibition T1 (pure Birds Nest 55mm fern extract) 54mm 56mm 165mm 55mm SENSITIVE

T2 (5 ml Birds Nest 0mm fern w/ 5 ml H20) "#ntr#l ("efur#xime)

2mm

0mm

2mm

0 6!mm/ undefined

RESISTANT

46mm

50mm

4$mm

144mm

4$mm

SENSITIVE (%&' B(%&%)

Positi"e ;A< for => N sensiti"e 7egati"e ;5< for => Nresistant able9 sho+s that the result of the sensiti"ity test on Escherichia coli after $@ hours of incubation and e2posure to (ird6s 7est fern e2tract and the control group on its first trial' reat)ent 9 +hich is a pure (ird6s 7est fern e2tract has a )ean /one of inhibition of ?? )) obtained fro) the /one of inhibition of its replicates0 R9N??)), R$N?@)) and R:N ?B))' treat)ent $, a solution +ith bird6s nest fern plus +ater e2tract has a /one of inhibition of %'BC)) +ith replicates0 R9N%)), R$N$)) and R:N%))' treat)ent : ;Cefuro2i)e<, the control group, has a )ean rate of @M )) obtained fro) the )ean rate of its replicates0 R9N@B)), R$N?%)) and R:N@M))' (ased on the table 9, it +as found out that Escherichia coli applied +ith treat)ent 9 got the highest /one of inhibition and re)ar,ed Isensiti"e8, +hich i)plies a good and effecti"e antibacterial effect' Moreo"er, it is not directly co)parable to the control group since Cefuro2i)e is )anufactured for hu)an dosage +hile (ird6s 7est fern e2tract intended for direct inoculation to the bacteria' Con"ersely, treat)ent $ ;bird6s nest fern +P H$%< sho+ed the least => and therefore re)ar,ed as Iresistant8'

able $' Results of the sensiti"ity for the second trial'


R1 Treatments R2 R3 Total Mean Remarks Zone Of Inhibition 6*mm 6!mm

T1 (pure )irds nest 65mm fern extract)

201mm

6!mm

SENSITIVE

T2 (5 ml )irds nest 0mm fern w/ 5 ml H20)

0mm

0mm

0mm

0mm/ undefined

RESISTANT

Control ("efur#xime)

55mm

66mm

5*mm

1$0mm

60mm

SENSITIVE (%&' B(%&%)

Positi"e ;A< for => N sensiti"e 7egati"e ;5< for => Nresistant able$ sho+s that the result of the sensiti"ity test on Escherichia coli after $@ hours of incubation and e2posure to (ird6s 7est fern e2tract and the control group on its second trial' reat)ent 9 +hich is a pure bird6s nest fern e2tract has a )ean /one of inhibition of BC)) obtained fro) the /one of inhibition of its replicates0 R9NB?)), R$NB&)) and R:N BC))' reat)ent $, a solution +ith bird6s nest fern plus +ater e2tract has a /one of inhibition of %)) +ith replicates0 R9N%)), R$N%)) and R:N%))' reat)ent : ;Cefuro2i)e<, the control group, has a )ean rate of B%)) obtained fro) the )ean rate of its replicates0 R9N??)), R$NBB)) and R:N?&))' (ased on the table $, it +as found out that Escherichia coli applied +ith treat)ent 9 got the highest /one of inhibition and re)ar,ed Isensiti"e8, +hich i)plies a good and effecti"e antibacterial effect' Moreo"er, it is not directly co)parable to the control group since Cefuro2i)e is )anufactured for hu)an dosage +hile bird6s nest fern e2tract intended for direct inoculation to the bacteria' Con"ersely, treat)ent $ ;bird6s nest fern +P H$%< sho+ed the least => and therefore re)ar,ed as Iresistant'8 able :' Results of the sensiti"ity testing for the third trial'

R1 Treatments

R2

R3 Total Mean Remarks

Zone Of Inhibition 56mm 5$mm

T1 (pure )irds nest 55mm fern extract)

16*mm

56 +4mm

SENSITIVE

T2 (5 ml )irds nest +mm fern w/ 5 ml H20)

1mm

0mm

4mm

1 +4mm

RESISTANT

Control ("efur#xime)

45mm

4+mm

4$mm

1+6mm

45 +4mm

SENSITIVE (%&' B(%&%)

Positi"e ;A< for => N sensiti"e 7egati"e ;5< for => Nresistant able : sho+s that the result of the sensiti"ity test on Escherichia coli after $@ hours of incubation and e2posure to (ird6s 7est fern e2tract and the control group on its third trial' reat)ent 9 +hich is a pure bird6s nest fern e2tract has a )ean /one of inhibition of ?B':@)) obtained fro) the /one of inhibition of its replicates0 R9N??)), R$N?B)) and R:N ?M))' reat)ent $, a solution +ith bird6s nest fern plus +ater e2tract has a /one of inhibition of 9':@)) +ith replicates0 R9N:)), R$N9)) and R:N%))' treat)ent : ;Cefuro2i)e<, the control group, has a )ean rate of @?':@)) obtained fro) the )ean rate of its replicates0 R9N@?)), R$N@:)) and R:N@M))' (ased on the table :, it +as found out that Escherichia coli applied +ith treat)ent 9 got the highest /one of inhibition and re)ar,ed Isensiti"e8, +hich i)plies a good and effecti"e antibacterial effect' Moreo"er, it is not directly co)parable to the control group since Cefuro2i)e is )anufactured for hu)an dosage +hile bird6s nest fern e2tract intended for direct inoculation to the bacteria' Con"ersely, treat)ent $ ;bird6s nest fern +P H$%< sho+ed the least => and therefore re)ar,ed as Iresistant'8

able @' Analysis of Dariance of the first trial'


%#urce #f df ,ariance 'eplicati#n Treatment 1rr#r T#tal %% -% . /,alue . /ta) 50 6 *4 6 *4 10 1$ 00 1$ 00

QQSignificant Q7ot Significant (ased on the analysis of "ariance, the co)puted #re*uency "alue obtained for replication is RR' he tabular fre*uency ;$,@<NB'&@ at %'? and 9M at %'9' Hence, the replication is not significant' his )eans that replicates on the second trial do not really differ +ith each other' #or treat)ent, the co)puted f5"alue obtained is RRRRRR and the tabular #;$,@<NB'&@ at %'?le"el and 9M at %'9 le"el of probability, the treat)ent is significant +hich )eans that treat)ents really differ +ith each other in ter)s of /one of inhibition' able ?' Analysis of Dariance of the second trial'
%#urce #f df ,ariance 'eplicati#n Treatment 1rr#r T#tal %% -% . /,alue . /ta) 50 6 *4 6 *4 10 1$ 00 1$ 00

QQSignificant Q7ot Significant (ased on the analysis of "ariance, the co)puted #re*uency "alue obtained for replication is RR' he tabular fre*uency ;$,@<NB'&@ at %'? and 9M at %'9' Hence, the replication is not significant' his )eans that replicates on the second trial do not really differ +ith each other' #or treat)ent, the co)puted f5"alue obtained is RRRRRR and the tabular #;$,@<NB'&@ at %'?le"el and 9M at %'9 le"el of probability, the treat)ent is significant +hich )eans that treat)ents really differ +ith each other in ter)s of /one of inhibition' able B' Analysis of Dariance of the third trial'
%#urce #f df ,ariance 'eplicati#n Treatment 1rr#r T#tal %% -% . /,alue . /ta) 50 6 *4 6 *4 10 1$ 00 1$ 00

QQSignificant Q7ot Significant

(ased on the analysis of "ariance, the co)puted #re*uency "alue obtained for replication is RR' he tabular fre*uency ;$,@<NB'&@ at %'? and 9M at %'9' Hence, the replication is not significant' his )eans that replicates on the third trial do not really differ +ith each other' #or treat)ent, the co)puted f5"alue obtained is RRRRRR and the tabular #;$,@<NB'&@ at %'?le"el and 9M at %'9 le"el of probability, the treat)ent is significant +hich )eans that treat)ents really differ +ith each other in ter)s of /one of inhibition' able C' 3eneral tabulation of the trials conducted'
R1 Treatments R2 R3 Total Mean Remarks Zone Of Inhibition 6!mm 56 +4mm

T1 (pure )irds nest 55mm fern extract)

1!$ +4mm

5* 45mm

SENSITIVE

T2 (5 ml )irds nest 0 6!mm fern w/ 5 ml H20)

0mm

1 +4mm

2 01mm

0 6!mm

RESISTANT

Control ("efur#xime)

4$mm

60mm

45 +4mm

15+ +4mm

51 11mm

SENSITIVE (%&' B(%&%)

Positi"e ;A< for => N sensiti"e 7egati"e ;5< for => Nresistant able C sho+s the general tabulation of the trials conducted for the sensiti"ity of Escherichia coli to different treat)ents used' rials of treat)ent 9 +hich is a pure bird6s nest fern e2tract has a )ean /one of inhibition of ?&'@?)) obtained fro) the /one of inhibition of its replicates0 rial9N??)), rial $NBC)) and rial :N ?B':@))' reat)ent $, a solution +ith bird6s nest fern plus +ater e2tract has a /one of inhibition of %'BC)) +ith replicates0 rial 9N%'BC, rial $N%)) and rial :N9':@))' reat)ent : ;Cefuro2i)e<, the control group, has a )ean /one of inhibition of ?9'99)) obtained fro) the )ean of its trials0 rial 9N@M)), rial $NB%)) and rial :N@?':@))' (ased on the table C, it +as found out that Escherichia coli applied +ith treat)ent 9 got the highest /one of inhibition and re)ar,ed Isensiti"e8, +hich i)plies a good and effecti"e antibacterial effect' Moreo"er, it is not directly co)parable to the control group since Cefuro2i)e is )anufactured

for hu)an dosage +hile (ird6s 7est fern e2tract is intended for direct inoculation to the bacteria' Con"ersely, treat)ent $ ;bird6s nest fern +P H$%< sho+ed the least => and therefore re)ar,ed as Iresistant'8 able M'3eneral A7.DA
%#urce #f ,ariance df 'eplicati#n Treatment 1rr#r T#tal %% -% . /,alue . /ta) 50 6 *4 6 *4 10 1$ 00 1$ 00

QQSignificant Q7ot Significant (ased on the Analysis .f Dariance, the co)puted #re*uency "alue obtained for trial is RR' he tabular fre*uency ;$,@<NB'&@ at %'? and 9M at %'9' Hence, the trial are not significant' his )eans that result of the trial do not really differ +ith each other' #or treat)ent, the co)puted f5"alue obtained is RRRRRR and the tabular #;$,@<NB'&@ at %'?le"el and 9M at %'9 le"el of probability, the treat)ent is significant +hich )eans that treat)ents really differ +ith each other in trials conducted in ter)s of /one of inhibition' Statistically, the analysis of "ariance of the first, second and third trial sho+ed al)ost the sa)e results' >t therefore i)plies that the study is "alid' Results abo"e sho+ed high sensiti"ity of bacteria Escherichia coli to pure bird6s nest fern e2tract as sub4ected to Kirby5(auer Antibiotic esting'

CHA,)ER !
S9((AR< $F F"N "N'S= C$NC&9S"$N= AN REC$((EN A)"$N his chapter discusses the findings, conclusions, and reco))endations of the research study on IAntimicrobial Activity of Birds Nest Ferns Extract Against Escherichia Coli>+ >t +as conducted at Sultan Kudarat Polytechnic State College5 Laboratory High School Research Center , EJC Montilla, acurong City fro) Septe)ber 9$59B $%%& using the Analysis of Dariance of Co)pletely Rando)i/ed !esign ;CR!< +ith three treat)ents replicated three ti)es' ?(a.or Finding he )a4or findings of this study +ere as follo+s0 9' reat)ent 9;pure birds nest e2tract< got a general )ean of its => +ith ?&'@? )) +hile reat)ent $ ;? )l bird6s nest e2tract A ?)l of

sterili/ed +ater< got a general )ean of %'BC))' he control cefuro2i)e got a general )ean of ?9'99))' $' Escherichia coli sho+ed high sensiti"ity +hen e2posed to pure bird6s nest fern e2tract' :' >t is therefore found out that the treat)ent 9 is +here Escherichia coli is )ost sensiti"e to' @' (ased on the Analysis of Dariance, treat)ents ha"e significant differences a)ong each other in all trials conducted, +hile the replicates and trials are not significant ?Concl/sions he follo+ing conclusions +ere based fro) the )a4or findings' (ased on the findings of the test conducted, bacteria Escherichia coli is fa"orably sensiti"e to treat)ent 9, +hich is co)posed of pure birds nest fern e2tract fro) its lea"es as it re)ar,ed sensiti"e' he treat)ents sho+ed highly significant differences +hile the replicates and trials are "ery precise and sho+ed not so )uch significance' Li,e+ise, the co)parison bet+een the control group and the treat)ents used cannot be directly )easured since the control ;i'e'Cefuro2i)e< is )anufactured considering the dosage for hu)an consu)ption +hile bird6s nest fern e2tract is )ade to co)bat E' coli in direct contact +ith the bacteria itself' 7e"ertheless, the study generally pro"ed the great potential of bird6s nest fern e2tract to be the radical antidote for the threatening public health night)ares, the E' coli' ?Recommendations Studies on the different )ediu)s had sho+n significant findings' >n this connection, the researchers strongly reco))end that0 9' he sensiti"ity testing using pure birds nest e2tract to another sa)ple of bacteria be conducted' $' !eter)ine the S>R legend for birds nest fern e2tract to deter)ine the accurate and precise co)parison +ith co))ercial antibiotics' :' #urther study should be conducted to )anufactured birds nest fern in e2act dosage intended for hu)an consu)ption'

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