Inhibitory Activity of Plumeria Rubra (Kalachuchi), Ipomea Quatica

UV Journal of Research 2014 26 UV Journal of Research 27 Inhibitory activity of Plumeria rubra (Kalachuchi), The table shows that the higher the percentage an effective antimicrobial agent’s against E. of the extract the lesser the contamination of coli Ipomea quatica Mimosa pudica constituents that are responsible for the death of Euphorbia hirta Coleus E. coli. causing diarrhea. The extract holds active the ring. This means that 100% solution has the aromaticus (Oregano)(Kangkong), plant extracts against greater possibility of inhibiting the bacteria. Table 4. Percentage of E.coli killed V. CONCLUSION (Makahiya),Staphylococcus aureus(Gatas-Gatas) coagulase production and Based on the data gathered, the leaf extract Percentages of Bacteria Killed of M. micrantha (Chinese creeper) is an effective Maria Feibe Pastoril Dosage Trial 1 Trial 2 Trial 3 Average College of Pharmacy University of Visayas

100 % 75.0% 95.3% 96.0% 88.76 activeantimicrobial constituents agent that against are E.responsible coli causing for 75% 49.0% 90.2% 91.3% 76.83 thediarrhea. death Theof E. leaf coli. extract The effectiveness contains different of the [email protected] 50% 30.6% 87.2% 87.2% 68.33 Date submitted:April 11, 2014 Date of acceptance: June 30, 2014 Tetracyclin antimicrobial activity of M. micrantha depends 25% 0.0% 79.3% 85.2% 54.83 Controlled 0 97.0% 97.0% 97.0% 97 higher the level of concentration the higher the ABSTRACT 0.0% 0.0% 0.0% The table shows the effectiveness of leaf on the level of concentration. This means that E.coli the antimicrobial effect of the leaf extract of M. P. rubra M. pudica amount of the bacteria killed. Furthermore, micrantha is comparable to the pharmaceutical E. hirta C. aromaticus extract concentration in killing . 100% Thus, this shows that the higher the concentration The leaf leaf extracts extracts of of P. rubra (KALACHUCHI),(Kalachuchi), I. I. aquatica aquatica (KANGKONG), (Kangkong),S. aureus M. pudica solution is more effective than the 25% solution. of M. micrantha may now be a promising source (Makahiya),(MAKAHIYA), E. hirta (Gatas-Gatas), (GATAS-GATAS), and and C. aromaticus (Oregano) (OREGANO) were screenedE. werehirta Furthermore, the M. micrantha leaf extract is indrug the which quest is for the new Tetracycline. antimicrobial The leavesdrugs extractdue to forscreened inhibitory for inhibitory activity against activity coagulase against coagulase production production of of S. . aureus. The Tube The the higher the amount of the bacteria killed. comparable to the pharmaceutical drug which is CoagulaseTube CoagulaseS. aureus Test and Test Colony and Colony Count Count were usedwere forused the for inhibitory the inhibitory assay. assay. The The E. (Gatas-Gatas),hirta (GATAS-GATAS), and C. aromaticus and C. aromaticus (Oregano) (OREGANO were found were to found have inhibitoryto have inhibitory activity its efficacy. against coagulase production both having a mean grade level of 1 and a originality index: 91 % vitroactivity tests against conducted S. aureus do not, coagulase in any way,production stimulate both the having complexity a mean of gradethe human level TableTetracycline. 5. Phytochemical testing of Mikania micrantha meanof 1 and colony a mean count colony of 193.33 count andof 193.33 229.67, and respectively. 229.67, respectively. However, the However, in vitro the tests in similarity index: 9 % E. hirta C. aromaticus conducted do not, in any way, stimulate the complexity of the human body. Instead, Active Actual Reagents Results Remarks paper id: 470233195 Constituets results thesebody. Instead,results warrant these results the warrant (Gatas-Gatas) the E. hirta and(GATAS-GATAS) and (Oregano) C. aromaticus plant Grammarly: checked Keywords:extracts(OREGANO) inhibitory to further activity,plant extractsanti-coagulase Staphylococcus to further investigation. aureus, anti-coagulase coagulase investigation. production, antibacterial, herbal plants Mayer’s Creamy Creamy + reagent color SaponinsAlkaloid Water Foam Foamy + REFERENCES I. INTRODUCTION of view, it is obvious that natural products, either Herbal plant is a vast wealth of nature not Fehling of view, it is obvious that natural products, either solutions Red Red + Ayensu, E.S. (1981). Medicinal plants of West Indies. only from the global environmental perspective plant extracts, provide unlimited opportunities to A and B Algonac, MI: Reference Publications. Herbal plant is a vast wealth of nature not in a form of pure compounds or as standardized Glycosides Blackwell, W.H. (1990). Poisonous and medicinal plants. only from the global environmental perspective plantThe extracts, increase provide in unlimiteddrug-resistant opportunities bacteria to Ferric green Tannins green + Englewood Cliffs, New Jersey: Prentice Hall. but also from the medicinal point of view. It develop a variety of new drugs. Chloride Darkor deep butresistant also abilityfrom the and medicinal combating point against of view.various It has pressed on the search for alternative and Dark DeFilipps, R.A., Maina, S.L., & Crepin, J. (2004). blue plays a significant role in improving the disease The increase in drug-resistant bacteria Medicinal plants of Guianas. Washington, DC: playsunfavorable a significant metabolic role in activities improving within the disease the ppt. Smithsonian Institution. resistant ability and combating against various has pressed on the search for alternative and Ferric green One potential source of antibiotics is plants Phenols green + natural sources of antibiotics (Saeed et al., 2005). Chloride Darkor deep Facey, B., Mulder, H.P.C., & Porter, R.B. (2004). unfavorable metabolic activities within the natural sources of antibiotics (Saeed et al., 2005). Dark blue Mikanolide from Jamaican Mikania livingremedies system. that have Numerous been proved infectious variously diseases since One potential source of antibiotics is plants micrantha. Acta Crystallographica Section C, Legend: (-) Negative; (+) Heavy precipitateppt. have been known to be controlled by herbal (Joshi et al., 2009). Plants such as Kalachuchi, 60(1), 798–800. are widespread species in the Philippines and Kangkong, Makahiya, Gatas-Gatas, and Oregano Li, Y., Li, J., Wang, X., & Cao, A. (2013). Antimicrobial remediesSince time that immemorial, have been man proven has variously used various since constituents of the leaves of Mikania micrantha. primitiveparts of plants to to present presentin treatment history history and of of theprevention mankind. of areMoreover, widespread plants species are not in only the Philippinesvery accessible and are used in traditional medicine in the country. testing of M. micrantha in the different reagents HBK PLoS ONE, 8(10): e76725. doi:10.1371/ Since time immemorial, man has used various andare usedeffective in traditional against disease-causing medicine in the microbes country. journal.pone.0076725 Moreover, plants are not only very accessible Table 5 shows the resultsM. micrantha of phytochemical has an partsunmatched of plants varieties in treatment of compounds and preventionare present ofin but also safer to use than commercial antibiotics Patamona, G. (2004). Medicinal plants of the Guianas. various ailments. Unimaginably, unrevealed and and effective against disease-causing microbes Guyana, Surinam, French Guiana. but also safer to use than commercial antibiotics showsused. It that further the leafshows extract that of M. micrantha has unmatched varieties of compounds are present in active chemical contituents. The result further the diversified herbs on earth. From thesethese pointspoints (Chaudhry et al., al., 2006). 2006). A A study study was shown 2828 UV Journal of Research 29

Pastoril, M. F. that extraction of the crude plant P.P. rubrarubra that forming, catalase-positive and facultatively nosocomially- acquired infections, being the III. MATERIALS AND METHODS contains iridoids that have been reported to anaerobic organism that belongs to the family most frequently isolated bacterial pathogen thathave extractionantibacterial, of the algicidal, crude plantcytotoxic, and/orthat forming, catalase-positive and facultatively contains iridoids that have been reported to anaerobic organism which belongs to the family pairs, and in irregular clusters that have been Although antibiotic agents are now available in This study utilizes experimental research have antibacterial, algicidal, cytotoxic, and/or Micrococceae.Micrococceae. They maymay be be found found singly, singly, in I.I. aquatica aquatica exerted a . from patients with hospital-acquired infections. onlydesign, one specifically single variable parallel group(control design group) where is plantplant growth growth inhibitory inhibitory activity activity (Kardono (Kardono et et al., al., in pairs, and in irregular clusters that have been high magnitude of antimicrobial activity against resistance of S. aureus is great due to its ability five groups were used at the same time with 1990).1990).the tested It It wasfour foundtypesfound of thatthat bacterial speciesexerted namely describedorganismsdescribed as asare “bunches “bunches resistant of ofto grapes” grapes”.temperatures The The cell cell as wallhigh wall the market, it cannot be denied that antibiotic EscherichiaaEscherichia high magnitude coli,coli, ofPseudomonasPseudomonas antimicrobial activityaeruginosa,aeruginosa, against S.S. containscontains peptidoglycan peptidoglycan and and teichoic teichoic acid. acid. The The manipulated (Calmorin and Calmorin, 2002). the tested four types of bacterial species: namely, organisms are resistant to temperatures as high aureusaureus and MicrococcusMicrococcus luteusluteus Colonies are usually large (6-8 mm in diameter), antimicrobialto form fibrin clotsubstances and thus with protecting inhibitory itself mode from obtainedIn this experiment, through decoction five different method herbalof extraction plant asas 50°C, 50°C, to to high high salt salt concentrations, concentrations, and and to to drying. drying. M.M. pudicapudica was found to exhibit in vitro . ofphagocytosis action may making have fewer it more side virulent. effects thanThe usethose of withextracts uniform were compared.amount of All grams of these of herbalextracts plants were and (Majumdar(Majumdar et et al., al., Colonies are usually large (6-8 mm in diameter), 2009).2009). was found to exhibit in vitro smooth,smooth, and and translucent translucent. ColoniesColonies appearappear creamy,creamy, ofof E. E. hirta hirta that were extracted by maceration in conducted at Pharmacy Laboratory and Medical bacteriostaticethanolbacteriostatic were activity usedactivity in (Genest traditional(Genest et et al., al.,medicine 2008). 2008). Leaves Leavesfor the S.white,S.white, aureusaureus or or light light gold gold and and “buttery “buttery looking” looking” after after with bactericidal mode of action. The latter ones and volume of solvent. The experiment was treatmentwhich of boils, were wounds extracted and control by maceration of diarrhea in 18but18 to toit 24 24may hours hours be foundof of incubation incubation regularly (Stoppler, (Stoppler, in most 2009). 2009).other tend to kill all of the bacteria in the body including ethanol were used in traditional medicine for the colonizes colonizes mainly mainly the the nasal nasal passages, passages, normal flora whereas the former ones just retard andTechnology laboratory Laboratory apparatuses, of all St. relevant Scholastica’s test treatment of boils, wounds and control of diarrhea but it may be found regularly in most other the growth of the bacteria which are further killed andCollege measurements, Tacloban. For such easy as accessweighing of and reagent the andand dysenterydysentery (Ogueke (Ogueke et et al.,al., 2007).2007). AA studystudy waswas anatomicalanatomical locales,locales, includingincluding thethe skin,skin, oraloral cavitycavity oil which contains carvacrol and thymol against by theThe immune Researchers response then of theaim body to inhibit(Doss etthat al., extraction of the plants were done at Pharmacy previouslypreviously conductedconducted onon thethe efficacyefficacy of of Oregano oregano andand gastrointestinal gastrointestinal tract. tract. They They causes causes range range S.S. aureus aureus andS. S. epidermidis epidermidis (Nostro et pimples, boils (furuncles), cellulitis folliculitis, coagulase2011). production of S. aureus oil which contains carvacrol and thymol against ofof illnesses illnesses from from minor minor skin skin infections infections such such as as P. rubra (KALACHUCHI), of S. aureus and performing Tube Coagulase Test planktonicplanktonic and (Nostro et pimples, boils (furuncles), cellulitis folliculitis, Antibiotic resistance has become a global and abscesses, to life-threatening diseases (2) I. aquatica using M.five pudicaherbal wereLaboratory. done at However, Medical Technology the inoculation, Laboratory culturing for al.,al., 2004). 2004). carbuncles,carbuncles, impetigo, impetigo, scalded scalded skin skin syndrome, syndrome, such as pneumonia, meningitis, osteomyelitis, plants. These are E. (1)hirta ), Antibiotic resistance becomes a global and abscesses, to life-threatening diseases antibiotics is being threatened by the emergence C. aromaticus (KANGKONG), (3) concern.concern. The The clinical clinical efficacy efficacy of of many many existing existing such as pneumonia, meningitis, osteomyelitis, (MAKAHIYA), (4) (GATAS-GATAS and (5) IV.safe, regulated,PREPARATION and conducive OF Henvironment.ERBAL PLANT antibiotics is being threatened by the emergence endocarditis,endocarditis, toxic toxic shock shock syndrome syndrome (TSS), (TSS), continuous and urgent need to discover new range from mild and requiring no treatment to II. OBJECTIVES (OREGANO). ExTRACTS ofof multidrug-resistant multidrug-resistant pathogens. pathogens. There There is is a a bacteremia,bacteremia, andand sepsis.sepsis. Staph-relatedStaph-related illnessillness cancan antimicrobial compounds with diverse chemical S.S. aureus aureus The study generally aimed to investigate the Plant Collection. P. rubra (KALACHUCHI), I. continuous and urgent need to discover new range from mild and requiring no treatment to structures and novel mechanisms of action primarily by the tube coagulase test (Larsen et inhibitory activity of P. rubra (KALACHUCHI), I. aquatica M. pudica antimicrobial compounds with diversed chemical severesevere and and potentially potentially fatal. fatal. is is identified identified S. aquatica M. pudica E. hirta ), and C. aromaticus structures and novel mechanisms of action primarily by the tube coagulase test (Larsen et Therefore, researchers are increasingly turning aureusof S. aureus E. hirta ), and C. aromaticus (KANGKONG), samples were collected(MAKAHIYA), from Happy forfor new new and and re-emerging re-emerging infectious infectious diseases. diseases. al.,al., 1995).1995). One important One important classification classification of The roleS. aureus of S. aureus (KANGKONG), (MAKAHIYA),S. aureus (GATAS-GATAS Therefore, researchers are increasingly turning is its is ability its ability to produce to produce coagulase. coagulase. The discoveries that lead to develop better drugs causes coagulation that allows the bacteria to (GATAS-GATAS individual(OREGANO) plant was randomly collected between theirtheir attentionattention toto folkfolk medicine,medicine, lookinglooking forfor newnew role of coagulasecoagulase is captivating. is captivating. It causes It aimed(OREGANO) to achieve plant the extractsfollowing: against Homes Diit, Tacloban City, Philippines. The discoveries that lead to develop better drugs coagulation that allows the bacteria to coat failure of chemotherapeutics and antibiotic . coagulase production. Specifically the researchers Collected samples were wrapped in clean plastic againstagainst microbial microbial infections. infections. The increasing The increasing failure itselfcoat itselfwith a with layer a of layer fibrin of under fibrin underwhich whichit hides it resistance exhibited by pathogenic microbial S. aureusS. aureus defense 1. Quantify the inhibitory effect of herbal plants bags8:00 and am transported to 10:00 am directly by uprooting to the Pharmacy method. of chemotherapeutics and antibiotic resistance fromhides the from immune the immune system making system it making more virulent it more infectious agents has led to the screening of several against coagulase production in S. aureus exhibited by pathogenic microbial infectious Coagulasevirulent. Coagulase is part of isthe part of the defense system medicinal plants for their potential antimicrobial 2. Compare the inhibitory activity of herbal samples were thoroughly washed with running agents has led to the screening of several medicinal (Todar,system 2011). (Todar, 2011). estimates that the mortality rate of S.S. aureusaureus plants in S. aureus coagulase production Laboratory for the preliminary procedures. The plants for their potential antimicrobial activity. TheThe World World Health Health Organization Organization (WHO) (WHO) several plants species exhibit promising 3. Identify herbal plants that have inhibitory Recentactivity. studies Recent have studies suggested have that suggested several plants that estimates that the mortality rate of S. aureusS. aureus is an opportunistic bacterial effect against S. aureus coagulase production water to remove debris. The plant materials were specieshave enormous exhibit promisingtherapeutic antimicrobial potential as they effects. can invasivepathogeninvasive infectionassociated was was about aboutwith 90% 90asymptomatic by% the by year the rinsed with distilled water. Each sample was Plant-basedantimicrobial antimicrobials effects. Plant-based have antimicrobials enormous year2011.2011. is an opportunistic bacterial serve the purpose with lesser side effects that are Null hypothesis. weighed 10 g. Only the healthy looking matured therapeutic potential as they can serve the pathogen associated with asymptomatic , difference on the inhibitory activity of P. rubra leavesExtraction in every herbal and Purification plant were picked of Plant and purpose with lesser side effects that are often colonizationcolonization of of the the skin skin and and mucosal mucosal surfaces surfaces Staphylococcus is a group of bacteria that of wound infections and has the potential to (KALACHUCHI), I. aquaticaThere is no significant M. Extract.randomly Decoctionselected to ofbe usedeach forherbal decoction. plant was associatedoften associated with synthetic with synthetic antimicrobials. antimicrobials. ofof normalnormal humans.humans. HoweverHowever, itit alsoalso isis thethe causecause can cause a number of diseases as a result of induce certain diseases, leading to infections pudica E. hirta ), and Staphylococcus is a group of bacteria that of wound infections and has the potential to infection of various tissues of the body. . S.S. aureusaureus C. aromaticus (KANGKONG), can cause a number of diseases as a result of induce certain diseases, leading to infections is a gram-positive cocci, non-motile, non-spore responsible for many serious community- and S. aureus(MAKAHIYA), (GATAS-GATAS containingprepared by the boiling leaves and 10g decoction of the leaves was removed in 50ml infection of various tissues of the body inin anyany ofof thethe majormajor organsorgans ofof thethe body.body. ItIt alsoalso isis (OREGANO) plant extracts against distilled water in a flask for 20 minutes. The flask is a gram-positive cocci, non-motile, non-spore responsible for many serious community- and coagulase production. 28 UV Journal of Research 29

Pastoril, M. F. that extraction of the crude plant P. rubra that forming, catalase-positive and facultatively nosocomially- acquired infections, being the III. MATERIALS AND METHODS contains iridoids that have been reported to anaerobic organism that belongs to the family most frequently isolated bacterial pathogen have antibacterial, algicidal, cytotoxic, and/or nosocomially-acquired infections, being the pairs, and in irregular clusters that have been mostAlthough frequently antibiotic isolated agents are bacterial now available pathogen in ThisThe study study utilizes experimental research Micrococceae. They may be found singly, in I. aquatica exerted a from patients, with hospital-acquired infections. onlydesign, one specifically single variable parallel group(control design group) where is plant growth inhibitory activity (Kardono et al., high magnitude of antimicrobial activity against Althoughresistance antibioticof S. aureus agents is great are due now to available its ability in five groups groups were were used used at theat the same same time withtime 1990).the tested It wasfour foundtypes of that bacterial species namely organismsdescribed asare “bunches resistant ofto grapes”.temperatures The cell as high wall the marketmarket, it cannot be denied that antibiotic with only one single variable (control group) is contains peptidoglycan and teichoic acid. The Escherichia coli, Pseudomonas aeruginosa, S. resistance of is great due to. its ability manipulated (Calmorin(Calmorin and & Calmorin, Calmorin, 2002). aureus and Micrococcus luteus Colonies are usually large (6-8 mm in diameter), toantimicrobial form fibrin clotsubstances and thus with protecting inhibitory itself mode from obtainedIn this experiment, through decoction five different method herbal of extraction plant as 50°C, to high salt concentrations, and to drying. M. pudica was found to exhibit in vitro phagocytosisof action may making have fewer it more side virulentvirulent. effects thanThe usethose of withextracts uniform were compared.amount of All grams of these of herbalextracts plants were (Majumdar et al., antimicrobial substances with inhibitory mode obtained through decoction method of extraction 2009). smooth, and translucent. Colonies appear creamy, of E. hirta that were extracted by maceration in of action may have fewer side effects than those conductedwith uniform at Pharmacy amount of Laboratory grams of herbal and Medical plants ethanolbacteriostatic were usedactivity in traditional(Genest et al.,medicine 2008). Leavesfor the S.white, aureus or light gold and “buttery looking” after with bactericidal mode of action. The latter ones and volume of solvent. The experiment was treatment of boils, wounds and control of diarrhea but18 toit 24may hours be found of incubation regularly (Stoppler, in most 2009).other tend to kill all of the bacteria in the body including conducted at Pharmacy Laboratory and Medical colonizes mainly the nasal passages, normal flora whereas the former ones just retard andTechnology laboratory Laboratory apparatuses, of St.all relevant Scholastica’s test the growth of the bacteria which are further killed andCollege measurements, Tacloban. For such easy as access weighing of and reagent the and dysentery (Ogueke et al., 2007). A study was anatomical locales, including the skin, oral cavity oil which contains carvacrol and thymol against by theThe immune Researchers response then of theaim body to inhibit(Doss etthat al., extractionand laboratory of the apparatuses,plants were done all relevant at Pharmacy tests previously conducted on the efficacy of oregano and gastrointestinal tract. They causes range S. aureus and S. epidermidis (Nostro et pimples, boils (furuncles), cellulitis folliculitis, 2011).coagulase production of S. aureus and measurements, such as weighing and the of illnesses from minor skin infections such as The researcher thenP.P. rubrarubra aim (KALACHUCHI), to inhibit that ofextraction S. aureus of and the performing plants were Tube done Coagulase at Pharmacy Test planktonic Antibiotic resistance has become a global and abscesses, to life-threatening diseases I.coagulase(2) aquatica I. aquatica production of M. pudica using M.five pudicaherbal wereLaboratory. done at However, Medical Technologythe the inoculation, Laboratory culturing for al., 2004). carbuncles, impetigo, scalded skin syndrome, such as pneumonia, meningitis, osteomyelitis, plants.E. hirta These are E. (1) (1)hirta ), (Kalachuchi),C. aromaticus), (2) and performing Tube Coagulase. Test antibiotics is being threatened by the emergence C. aromaticus(Kangkong),(KANGKONG), (3) (3) (Makahiya), were done at Medical Technology Laboratory for concern. The clinical efficacy of many existing (4)(MAKAHIYA), (Gatas-Gatas (4) (GATAS-GATASand (5) and (5) IV.safe, regulated,PREPARATION and conducive OF Henvironmentenvironment.ERBAL PLANT endocarditis, toxic shock syndrome (TSS), continuous and urgent need to discover new range from mild and requiring no treatment to II.(Oregano). OBJECTIVES (OREGANO). ExTRACTS of multidrug-resistant pathogens. There is a bacteremia, and sepsis. Staph-related illness can antimicrobial compounds with diverse chemical S. aureus The study generally aimed to investigate the Plant Collection.Collection. P.P. rubra rubra (KALACHUCHI), I. structures and novel mechanisms of action primarily by the tube coagulase test (Larsen et inhibitory activity of P. rubraP. rubra (KALACHUCHI), I. aquatica M. M. pudica pudica E. severe and potentially fatal. is identified aquaticaThe study generallyM. M. pudica aimedpudica to investigateE. E.hirta hirta ), C.), aromaticusand (Kalachuchi),C. aromaticus Therefore, researchers are increasingly turning of S. aureus E.thehirta hirtainhibitory activity), ofC. ), aromaticusand (Kalachuchi),C. aromaticus (KANGKONG),(Kangkong), samples were collected(Makahiya),(MAKAHIYA), from Happy for new and re-emerging infectious diseases. al., 1995). One important classification The role of S. aureus (KANGKONG),(Kangkong), S. aureus(makahiya),(MAKAHIYA), S. aureus (Gatas-Gatas(GATAS-GATASand (Oregano) is its ability to produce coagulase. discoveries that lead to develop better drugs causes coagulation that allows the bacteria to (Gatas-Gatas(GATAS-GATASand (Oregano) individual(OREGANO)samples wereplant was collected randomly from collected Happy between Homes their attention to folk medicine, looking for new coagulase is captivating. It aimed(OREGANO)plant to extracts achieve plant the against extractsfollowing: against coagulase HomesDiit, Tacloban Diit, Tacloban City, Philippines. City, Philippines. The individual The failure of chemotherapeutics and antibiotic coagulaseproduction. production. Specifically Specifically the researcher the researchers aimed to Collectedplant was samples randomly were collected wrapped between in clean 8:00 plastic am against microbial infections. The increasing coat itself with a layer of fibrin under which it resistance exhibited by pathogenic microbial S. aureus defense 1.achieve1. Quantify the following: the inhibitory effect of herbal plants bags8:00to 10:00 and am transportedto am 10:00 by uprooting am directly by uprooting method. to the Pharmacy Collected method. hides from the immune system making it more infectious agents has led to the screening of several against coagulase production in S. aureus;aureus samples were wrapped in clean plastic bags and virulent. Coagulase is part of the medicinal plants for their potential antimicrobial 2.2. QuantifyCompare thethe inhibitory inhibitory effect activity of herbal of herbalplants samplestransported were directly thoroughly to the washedPharmacy with Laboratory running system (Todar, 2011). estimates that the mortality rate of S. aureus plantsagainst in coagulase S. aureus productioncoagulase production in Laboratoryfor the preliminary for the preliminary procedures. procedures. The samples The The World Health Organization (WHO) several plants species exhibit promising 3.3. CompareIdentify herbal the inhibitory plants that activity have ofinhibitory herbal were thoroughly washed with running water to activity. Recent studies have suggested that S. aureus is an opportunistic bacterial effectplants against in S. aureuscoagulase coagulase production; production and waterremove to debris.remove The debris. plant The materials plant materials were rinsed were have enormous therapeutic potential as they can pathogeninvasive infectionassociated was aboutwith 90% asymptomatic by the year Identify herbal plants that have inhibitory rinsedwith distilled with distilled water. Each water. sample Each was sample weighed was antimicrobial effects. Plant-based antimicrobials 2011. serve the purpose with lesser side effects that are effectNull againsthypothesis. coagulase production weighed10 g. Only 10 theg. Only healthy the healthy looking looking matured matured leaves difference on the inhibitory activity of P. rubra leavesin everyExtraction in herbal every plant herbal and were Purification plant picked were and picked ofrandomly Plant and colonization of the skin and mucosal surfaces Staphylococcus is a group of bacteria that of wound infections and has the potential to (KALACHUCHI),I. aquaticaI. aquaticaThere is no significantM. pudica M. Extract.randomlyselected to Decoctionselected be used to for ofbe decoction. usedeach forherbal decoction. plant was often associated with synthetic antimicrobials. of normal humans. However, it also is the cause can cause a number of diseases as a result of induce certain diseases, leading to infections pudicadifference on E. the hirta inhibitory E. hirta activity ), of ), andC. infection of various tissues of the body. S. aureus C.(Kalachuchi),aromaticus aromaticus (Kangkong),(KANGKONG), S. Decoction of each herbal plant was is a gram-positive cocci, non-motile, non-spore responsible for many serious community- and S.(Makahiya),aureus aureus (MAKAHIYA), (Gatas-Gatas(GATAS-GATASand containingprepared by the boiling boiling leaves 10and 10g g decoction of the leaves leaves was removed in in 50 50ml ml in any of the major organs of the body. It also is (Oregano)(OREGANO) plant plant extracts extracts against against distilled water in a flask for 20 minutes. The flask coagulasecoagulase production. production. containing the leaves and decoction was removed 30 UV Journal of Research 31

Pastoril, M. F.

VI. STATISTICAL ANALYSIS mean of clot inhibition in tubes with plant extract The Tube Coagulase Test and MHA colony (experimental group) to the mean of tube without from the heat and allowed to cool. The content of of 11% % barium chloride (0.05ml) and 99% % sulfuric counting were done in triplicates and the mean flask was filtered through filter paper to obtain acid (9.95ml). McFarland Standard was stored values and standard error of the mean were clear decoction. inwhirled standing again position for at atleast 4°C 1to minute 8°C and to protected obtain a plant extract (control group) as significant or Then,Syringe each plant filter extracts with awere general stored size in separate of 0.45 from light during 12 weeks. The suspension was. main tool in this study for easy comparison of VII.nonsignificant. RESULTS AND DISCUSSIONS autoclavedmicron will was reagent be used used bottles to to purify purify and the werethe plantplant placed extracts. inside whirledA full loop again of bacterial for at least growth one minutewas obtained to obtain with a calculated. Tabular form representation was the The results in experiment 1 using Plasma Then, each plant. extract was stored in separate homogenousan inoculating suspension loop and placed and to in break a test the tube clumps with The researchers used one-way analysis of autoclavedTest for reagent Contaminants bottles and in was Plant placed Extract. inside A full loop of bacterial growth was obtained with results obtained. the refrigeratorrefrigerator. an inoculating loop and placed in a test tube with of clot inhibition in the Tube Coagulase Test and gradeX and level experiment and turbidity 2 using were Plasma measured Y in gradeafter presence of organisms in every plant extract and 33ml ml ofof NutrientNutrient Broth.Broth. TheThe testtest tube was whirled Colonyvariance Count (one-way results ANOVA) of the to analyzetubes with the meanplant level after 4 hours, no clot were formed. The wasTrypticase prepared Soy Agaraccording (TSA) to was the used manufacturer’s to check the forbacterial at at least least suspension’s 1 one minute minute toturbidity break to break thewas clumps theadjusted clumps until to extract as the experimental group with tube 1 untila fairly a fairly turbid turbid suspension suspension was was obtained. obtained. The containing P. rubra (Kalachuchi), tube 2 I. aquatic 24 hours. 10µl of the mixture in each tube were C bacterial suspension’s turbidity was adjusted to M. pudica ofobtained incubation, and immediately numerous streakedgrowths inof MHA colonies with instructions. A small sample of extract was° besuspension the same asto thereduce turbidity turbidity, of the 0.5while McFarland more 4 E. hirta C. aromaticus the use of bacterial cell spreader. After 24 hours inoculated ontointo TSA and was incubated at 37 standards. More broth was added to the bacterial (Oregano)(Kangkong), to tube the 3control group (Makahiya), with no plant tube V.overnight. PREPARATION OF BACTERIAL CULTURES suspensionThe bacterial to concentration reduce turbidity, of the while bacterial more (Gatas-Gatas) and tube 5 Thewere number seen. Thus, of colonythe amount growth of mixtureobtained streaked shown Isolation of S. aureus Strains. colonies were added 8 CFU/ml to increase (colony turbidity.turbidity. forming Nevertheless, Duncan’s Multiple Range in MHA was reduced to 0.1µl in experiment 3. of S. aureus were obtained from the Eastern The bacterial concentration8 of the bacterial extract. plasma used in experiment 3 was pooled plasma Stock cultures suspensionInhibitory was Assay1.5Assay xx10 10 of CFU/ml Herbalof Herbal(colony Plant Extract.formingPlant computation of numerical boundaries that allow on Table 1.2 was less than 300 cfu/ml. Also, the Extract.unit/unit/milliliter). milliliter). The inhibitoryP. rubra activity of the P. Test (DMRT) was also used. It involves the Visayas Regional Medical Center (EVRMC). The I.rubra aquatica (KALACHUCHI), I.M. aquatica pudica E. from A, B and C. stock cultures were maintained by sub-culturing° hirtaTheM. pudica inhibitory activity), of C. E.the aromaticushirta (Kalachuchi), ), for the classification of the difference between the in NutrientnutrientIdentification agarAgar media of and were Bacteria. incubated at 37 and C. aromaticus(Kangkong), (Makahiya),(KANGKONG), Table 1.2. Summary of Data Gathered on Grade Level of Clot Formation, Turbidity and Colony Count on the Inhibitory C overnight. determined(Gatas-Gatas(MAKAHIYA), by measuringand the grade(GATAS-GATAS level(Oregano) of the Assay Against S. aureus Coagulase Production Using Plasma X (Experiment 1) and Plasma Y (Experiment 2) morphological characteristics of the S. aureusGram’s plant extracts were (OREGANO) determined plant by measuring extracts were the Experiment No. 1 Experiment No. 2 staining Tube technique Coagulase was Test performed was done for to the check further the Sixgrade test level tubes of thewere inhibition prepared, of theeach clot tube in thecontains Tube S. aureus . inhibitionCoagulase of Test. the SixclotS. testin aureus the tubes Tube wereCoagulase prepared, Test. Tube Coagulase Test was done for the further each tube contains 0.5 ml human plasma and 0.1 Grade Level Grade Level S. aureus , Clot Formation Colony Clot Formation Colony Turbidity Turbidity confirmation for the presenceS. aureus. of Then, it. Onewas of0.5ml plant ofml standardized human extract,P. rubra plasma with tubeand 0.11 containing. ml Five of standardized tubes I.P. aquatic rubrawere Count Count 4 hrs 24 hrs 4 hrs 24 hrs tube was filled with C and 0.5 was ml observed of human up plasma to four (Kalachuchi),treated with. Five 0.1tube tubes ml 2M. ofI. werepudica aquaticplant treated extract with with 0.1tube ml 1 Herbal Plant Extract with EDTA and ° 0.1 ml of Then, it was containing (Kalachuchi), tube 2 E.3 M.hirta pudica E.C. hirta aromaticus P. rubra No clot +1 Less Turbid TNTC No clot +2 Less Turbid TNTC (KALACHUCHI) incubated at 37 C and was observed up to four (Kangkong), tube 3 C. aromaticus(Makahiya), (Kangkong), tube tube C4 hours and 24 hours after incubation. The grade (Gatas- (Makahiya), Gatas) and tube tube 4 5 (Gatas-° the complete volume occupied by the broth; (3 C and observed up to I. aquatica No clot +1 Less Turbid TNTC No clot +3 Less Turbid TNTC level waswas measuredmeasured asas (4 (4 +) +) if if the the fibrin fibrin clot clot filled fills Gatas)(Oregano). and Alltube tubes 5 were° incubated (Oregano). at 37 All the complete volume occupied by the broth; (3 tubesand observed were incubated up to four at 37 hours and 24 hours of total volume occupied by the broth; (2 +) if the left untreated and served as basis for comparison (KANGKONG)M. pudica No clot +2 Less Turbid TNTC No clot +2 Less Turbid TNTC +) if the clot fills more than half but less than the 4incubation. hours and Tube 24 hours six was of left incubation. untreated Tube and served6 was total volume occupied by the broth; (2 +) if the as basisIn every for plate comparison of Mueller on Hinton the grade Agar level (MHA), for (MAKAHIYA) E. hirta No clot +1 Turbid TNTC No clot +3 Turbid TNTC clot formation;fills less than (negative) half the if total no clot volume observed occupied but a oncoagulase the grade production. level for coagulase production. by the broth; (1 +) if there is a little disorganized In every plate of Mueller Hilton Agar (MHA), bylittle the amorphous broth; (1 +) deposit if there might is a little be seen disorganized (Spencer C.(GATAS-GATAS) aromaticus clot formation; (negative) if no clot observed but a 0.1µl0.1 µl ofof thethe mixturemixture inin eacheach tube were obtained No clot +1 Turbid TNTC No clot +3 Turbid TNTC little amorphous deposit might be seen (Spencer and immediately streaked with the use of bacterial Standardization of the Bacteria in this (OREGANO)Conrol group No clot +3 Less Turbid TNTC No clot +3 Less Turbid TNTC study.&and Tatini, Tatini, 1974). 1974). havecell spreader.the same Growth number of colony colonies growth in each as to MHA the plate was counted to check if the treated bacteria in a liquid0.5 suspension McFarland with Standard a chemical was usedsolution to have the same number of colony growth as to the standardize the approximate number of bacteria untreated bacteria. in a liquid suspension with a chemical solution 30 UV Journal of Research 31

Pastoril, M. F.

VI. STATISTICAL ANALYSIS mean of clot inhibition in tubes with plant extract The Tube Coagulase Test and MHA colony (experimental group) to the mean of tube without from the heat and allowed to cool. The content of of 1% barium chloride (0.05ml) and 9% sulfuric counting were done in triplicates and the mean flask was filtered through filter paper to obtain acid (9.95ml). McFarland Standard was stored valuesThe and Tube standard Coagulase error Test of andthe MHAmean colony were clear decoction. whirledin standing again position for at atleast 4°C 1to minute 8°C and to protected obtain a counting were done in triplicates and the mean plant extract (control group) as significant or Then,Syringe each plant filter extracts with awere general stored size in separate of 0.45 from light during 12 weeks. The suspension was. valuesmain tool and in standardthis study error for easy of the comparison mean were of VII.nonsignificant. RESULTS AND DISCUSSIONS autoclavedmicron will reagent be used bottles to purify and werethe plant placed extracts. inside A full loop of bacterial growth was obtained with calculated. Tabular form representation was the The results in experiment 1 using Plasma anhomogenous inoculating suspension loop and placed and to in break a test the tube clumps with mainThe tool researchers in the study used for one-way easy comparison analysis of Test for Contaminants in Plant Extract. results obtained. The results in experiment 1 using Plasma the refrigerator. of clotThe inhibition researcher in the used Tube one-way Coagulase analysis Test and of gradeX and level experiment and turbidity 2 using were Plasma measured Y in gradeafter presence of organisms in every plant extract and 3ml of Nutrient Broth. The test tube was whirled varianceColony Count (one-way results ANOVA) of the to analyzetubes with the mean plant level after 4 hours, no clot were formed. The wasTrypticase prepared Soy Agaraccording (TSA) to was the used manufacturer’s to check the bacterialfor at least suspension’s 1 minute toturbidity break thewas clumps adjusted until to ofextract clot inhibitionas the experimental in the Tube groupCoagulase with Test tube and 1 grade level and turbidity were measured after a fairly turbid suspension was obtained. The Colonycontaining Count PP.. rubra results (Kalachuchi), of the tubes tube with2 I. aquatic plant 24 hours. 10µl10 µl of of the the mixture mixture in in .each each tube tube were was C extract as the experimentalM. pudica group with tube 1 ofobtained incubation, and immediately numerous streakedgrowths inof MHA colonies with instructions. A small sample of extract was° suspensionbe the same asto thereduce turbidity turbidity, of the 0.5while McFarland more containing4 E. hirta (Kalachuchi), tubeC. aromaticus2 the use of bacterial cell spreaderspreader. After 24 hours inoculated onto TSA and was incubated at 37 standards. More broth was added to the bacterial (Kangkong),(Oregano) to tube the 3control group (Makahiya), with no plant tube of incubation, numerous growths of colonies V.overnight. PREPARATION OF BACTERIAL CULTURES The bacterial concentration of the bacterial 4 . (Gatas-Gatas)(Gatas-Gatas) andand tubetube 55 Thewere number seen. Thus, of colonythe amount growth of mixtureobtained streaked shown Isolation of S. aureus Strains. colonies were added 8 CFU/ml to increase (colony turbidity. forming (Oregano)Nevertheless, to the controlDuncan’s group Multiple with no Range plant in MHA was reduced reduced to to 0.1 0.1µl µl in experiment 3. of S. aureus were obtained from the Eastern extractextract. plasmaThe number used in of experiment colony growth 3 was obtained pooled plasma shown Stock cultures suspensionInhibitory was 1.5Assay x10 of Herbal Plant computation of numerical boundaries that allow on Table 1.2 was less than 300 cfu/ml. Also, the Extract.unit/ milliliter). The inhibitory activity of the P. Test (DMRT) was also used. It involves the plasma used in experiment 3 was pooled plasma Visayas Regional Medical Center (EVRMC). The rubra (KALACHUCHI), I. aquatica from A, B and C. stock cultures were maintained by sub-culturing° M. pudica E. hirta ), for the classification of the difference between the in NutrientIdentification Agar media of and were Bacteria. incubated at 37 and C. aromaticus (KANGKONG), Table 1.2. Summary of Data Gathered on Grade Level of Clot Formation, Turbidity and Colony Count on the Inhibitory Summary of data gathered on Grade Level of Clot Formation, Turbidity and Colony Count on the Inhibitory Assay against S. aureus Coagulase Production Using Plasma X (Experiment 1) and Plasma Y (Experiment 2) C overnight. determined(MAKAHIYA), by measuring the grade(GATAS-GATAS level of the Assay Against S. aureus Coagulase Production Using Plasma X (Experiment 1) and Plasma Y (Experiment 2) morphological characteristics of the S. aureusGram’s (OREGANO) plant extracts were Experiment No. 1 Experiment No. 2 staining Tube technique Coagulase was Test performed was done for to the check further the Six test tubes were prepared, each tube contains S. aureus . inhibition of the clot in the Tube Coagulase Test. Grade Level Grade Level S. aureus Clot Formation Colony Clot Formation Colony Turbidity Turbidity confirmation for the presenceS. aureus. of Then, it. Onewas of0.5 plant ml human extract, plasma with tubeand 0.11 containing ml of standardized P. rubra Count Count 4 hrs 24 hrs 4 hrs 24 hrs tube was filled withC and 0.5 was ml observed of human up plasmato four (Kalachuchi),. Five tube tubes 2 I. wereaquatic treated with 0.1 ml Herbal Plant Extract with EDTA and °0.1 ml of P. rubra 3 M. pudica E. hirta P. rubra No clot +1 Less Turbid TNTC No clot +2 Less Turbid TNTC (KALACHUCHI) incubated at 37 C. aromaticus (Kangkong), tube hours and 24 hours after incubation. The grade (Makahiya), tube 4 (Gatas- I. aquatica the complete volume occupied by the broth; (3 C and observed up to (Kalachuchi)I. aquatica No clot +1 Less Turbid TNTC No clot +3 Less Turbid TNTC level was measured as (4 +) if the fibrin clot fills Gatas) and tube 5 ° (Oregano). All tubes were incubated at 37 M. pudica total volume occupied by the broth; (2 +) if the left untreated and served as basis for comparison (Kangkong)(KANGKONG)M. pudica No clot +2 Less Turbid TNTC No clot +2 Less Turbid TNTC +) if the clot fills more than half but less than the 4 hours and 24 hours of incubation. Tube 6 was In every plate of Mueller Hinton Agar (MHA), E.(Makahiya)(MAKAHIYA) hirta E. hirta No clot +1 Turbid TNTC No clot +3 Turbid TNTC clot formation;fills less than (negative) half the if total no clot volume observed occupied but a on the grade level for coagulase production. by the broth; (1 +) if there is a little disorganized little amorphous deposit might be seen (Spencer (Gatas-Gatas)C.(GATAS-GATAS) aromaticus 0.1µl of the mixture in each tube were obtained No clot +1 Turbid TNTC No clot +3 Turbid TNTC and immediately streaked with the use of bacterial Standardization of the Bacteria in this (Oregano)(OREGANO)Conrol group No clot +3 Less Turbid TNTC No clot +3 Less Turbid TNTC study.and Tatini, 1974). havecell spreader.the same Growthnumber of colony colonies growth in each as to MHA the plate was counted to check if the treated bacteria in a liquid0.5 suspension McFarland with Standard a chemical was usedsolution to standardize the approximate number of bacteria untreated bacteria. 32 UV Journal of Research

Table 1.2. Summary of Data Gathered on Experiment No.3 Grade Level of Clot Formation, Turbidity and Colony Count 32 UV Journal of Research 32on the Inhibitory Assay Against S. aureus Coagulase Production Using Pooled Plasma (A,B,C)UV Journal of Research 33

Pastoril, M. F. Experiment No. 3 Table 1.2. Summary of Data Gathered on Experiment No.3 Grade Level of Clot Formation, Turbidity and Colony Count Summary of data gatheredTrial 1 on Experiment No. 3 GradeTrial Level 2 of Clot Formation, TurbidityTrial and 3 Colony Count with the mean of 2 did not show any inhibitory onon the Inhibitory Assay Against S. the Inhibitory AssayGrade against Level S. aureusaureus Coagulase Coagulase Production Using Pooled Plasma (A,B,C) ProductionGrade Level Using Pooled Plasma (A,B,C)Grade Level plant extracts against coagulase production activity against clot formation since it was the Herbal Plant Extract Clot Formation Colony Clot Formation Colony Clot Formation Turbidity Turbidity Turbidity The observed clot formation of the five herbal Count Count Colony of S. aureus may attribute to the compound it same as the mean of the control group based on 4 hrs 24 hrs 4 hrs 24 hrs 4 hrs 24 hrs Count Experiment No. 3 E. hirta (GATAS-GATAS) and C. Trial 1Less Trial 2Less Trial 3 P. rubra No clot +1 12 No clot +2 No clot +1 Less Turbid 24 (KALACHUCHI) TrialTurbid 1 TrialTurbid 2 Trial 3 aromaticus not prove that the plant extracts inhibit only the Grade Level Grade Level Grade Level possesses. The Table 2.1 data. The results for clot formation do I. aquatica Less Less 7 greater effect in inhibiting clot formation with S. aureus coagulase production thus observation Herbal Plant Extract NoClot clot Formation +1 Colony NoClot clot Formation +2 Colony110 NoClot clot Formation +1 Less Turbid 132 Grade Level TurbidityTurbid Grade Level TurbidityTurbid Grade Level Turbidity (OREGANO) were found to have Herbal Plant Extract Clot Formation ColonyCount Clot Formation ColonyCount Clot Formation Colony P. rubra (KALACHUCHI) and on turbidity and colony count were performed to 4 hrs 24 hrs TurbidityLess 79 4 hrs 24 hrs TurbidityLess 4 hrs 24 hrs Turbidity Count (KANGKONG)M. pudica No clot +2 Count No clot +2 Count26 No clot +2 Less Turbid Colony 4 hrs 24 hrs Turbid 4 hrs 24 hrs Turbid 4 hrs 24 hrs Count I. aquatica Less Less P. rubra No clot +1 12 No clot +2 No clot +1 Less Turbid 24 the mean of 1. The (KALACHUCHI)E. hirta (MAKAHIYA) Turbid 129 Turbid 94 in terms of inhibiting the clot formation with P.P. rubra No clot +1 TurbidLess No clot +1 TurbidLess 246 No clot +1 Turbid 181 No clot +1 Less 12 No clot +2 Less 7 No clot +1 Less Turbid 24 (KANGKONG) showed the same effect further investigate inhibitory activity of the five (KALACHUCHI)I. aquatica No clot +1 Turbid No clot +2 Turbid 110 No clot +1 Less Turbid 132 M. pudica C. aromaticus Turbid 153 Turbid (GATAS-GATAS) No clot +1 Turbid 228 No clot +1 Turbid No clot +1 Turbid 163 herbal plant extracts. (Kalachuchi)I.I. aquaticaaquatica Less 79 Less 7 (KANGKONG)M. pudica No clot +1+2 Less No clot +2 Less 11026 No clot +1+2 Less Turbid 132 Conrol group No clot +2 Turbid 112 No clot +2 Turbid 298123 No clot +2 Less Turbid the Table mean 2.2. of Turbidity of Five Different Plant Extracts and Control Group in Tube Coagulase Test 1.33. While (MAKAHIYA) (OREGANO) Turbid Turbid (MAKAHIYA) Less 12979 Less 94 M.(Kangkong)(KANGKONG)E. hirtapudica No clot +2 No clot +2 26 No clot +2 Less Turbid M. pudica No clot +1Turbid No clot +1Turbid 246 No clot +1 Turbid 181149 C. aromaticus(Makahiya) 153 E.E.(GATAS-GATAS) hirtaThe (MAKAHIYA) results Noin clot experiment +1 Turbid 1 using129 228 Plasma No clot The +1 number Turbid of colonyNo clot growth +1 obtained Turbid shown 16394 Replication No clot +1 Turbid No clot +1 Turbid 246 No clot +1 Turbid 181 Herbal Plant Extract Less Less 298 (OREGANO)Conrol group No clot +2 112 No clot +2 123 No clot +2 Less Turbid C.C. aromaticus Turbid 153 Turbid (Gatas-Gatas)(GATAS-GATAS) No clot +1 Turbid 228 No clot +1 Turbid No clot +1 Turbid 163 Trial 1 Trial 2 Trial 3 plasma used in experiment 3 was pooled plasma149 Less Less P. rubra (KALACHUCHI) Less Turbid Less Turbid Less Turbid X(Oregano)Conrol and group experimentNo clot 2 using +2 Plasma Y112 in grade No clot on +2 Table 1.2 was298123 less No clotthan 300 +2 cfu/ml. Less Turbid Also, the grade(OREGANO)The level results and inturbidity experiment wereTurbid 1measured using Plasma after The numberTurbid of colony growth obtained shown level after 4 hours, no clot were formed. The I. aquatica Less Turbid Less Turbid Less Turbid 149 levelfrom A,of B clot and C.formation of experimental group M. pudica Less Turbid Less Turbid Less Turbid The results in experiment 1 using Plasma Theplasma number used inof experimentcolony growth 3 was obtained pooled plasmashown (KANGKONG) X24 andhours. experiment 10µl of the 2 mixture using Plasma in each Y tube in gradewere on TableThe first 1.2 partwas dealsless than with 300 measuring cfu/ml. theAlso, grade the E. hirta (GATAS-GATAS) Turbid Turbid Turbid grade level and turbidity were measured after and control group 24 hours of observation and (MAKAHIYA) oflevelobtained incubation, after and 4 immediately hours, numerous no clot streakedgrowths were in formed.of MHA colonies with The counting the number of colonies formed in MHA C. aromaticus Turbid Turbid Turbid the use of bacterial cell spreader. After 24 hours fromplasma A, Bused and in C. experiment 3 was pooled plasma The first part deals with measuring the andforlevel in countingof each clot test formation the solution number of including experimental of colonies the formed controlgroup Conrol group Less Turbid Less Turbid Less Turbid grade24X andhours. level experiment 10µl and of turbidity the 2 mixture using were Plasma in measuredeach Y tube in gradewereafter on TableThe first 1.2 partwas dealsless than with 300 measuring cfu/ml. theAlso, grade the (OREGANO) grade level of clot formation of experimental inand MHA control for ingroup each 24 test hours solution of observation including andthe ofwereobtainedlevel incubation, seen. after and Thus, 4 immediately hours, numerous the amount no clot streakedgrowths of were mixture in formed.of MHA streakedcolonies with The counting the number of colonies formed in MHA The P. rubra (KALACHUCHI), I. aquatica that the coagulase production of S. aureus was thegroup use and of controlbacterial group cell spreader. 24 hours Afterof observation 24 hours fromcontrol A, Bgroup and C. and also observing its turbidity. Tablein MHA 2.1. wasGradeGrade Level of Clot Formation of Five Different Plant Extracts and Control Group in Tube Coagulase Test reduced Level of Clot to 0.1µlFormation in experiment of five different 3. plantlevelforgroup inextracts ofandeach clot alsoand test formation controlobserving solution group of its inTube including turbidity.experimental Coagulase the Test controlgroup M. pudica 24 hours. 10µl of the mixture in each tube were and Thecontrol first grouppart deals 24 hours with measuring of observation the grade and group showed the same characteristic in terms of obtainedwere seen. and Thus, immediately the amount streaked of mixture in MHA streaked with turbidity(KANGKONG) which is less turbid (MAKAHIYA) compared and to ControlE. hirta designatesinhibited orthe the presence bacteria of more were bacteria killed by in the of incubation, numerous growths of coloniesReplication counting the number of colonies formed in MHA inTable MHA 2.1. wasGrade Level of Clot Formation of Five Different Plant Extracts and Control Group in Tube Coagulase Test reduced to 0.1µl in experiment 3. group and also observingTreatment its turbidity. Treatment (GATAS-GATAS) and C. aromaticus solutionpresence and of that the the herbal bacteria plant did extracts. not form Turbid a clot the use Herbalof bacterial Plant cellExtract spreader. After 24 hours for in each test solutionTotal including theMean control (T) (x̅ ) Trial 1 Trial 2 Trial 3 (OREGANO) Replication were seen. Thus, the amount of mixture streaked Treatment Treatment showed in Table 2.2. Less turbid either indicates thus making the solution turbid. P. rubraHerbal (KALACHUCHI) Plant Extract +1 +2 +1 Total4 Mean Table 2.3. Colony Count (CFU/ml) of Five Different Plant Extracts and Control Group in Tube Coagulase Test inTable MHA 2.1. wasGrade Level of Clot Formation of Five Different Plant Extracts and Control Group in Tube Coagulase Test reduced to 0.1µl in experimentTrial 1 3. Trialgroup 2 and Trial also 3 observing(T) its turbidity. (x̅ ) I. aquatica(Kalachuchi) +1 +2 +1 4 1.33 Replication Treatment Treatment P. rubra (KALACHUCHI) +1 +2 +1 4 Herbal Plant Extract Total Mean M. pudica (Kangkong)(KANGKONG) +2 +2 +2 6 1.332 (T) (x̅ ) Replication Treatment Treatment Trial 1 Trial 2 Trial 3 I. aquatica +1 +2 +1 4 1.33 E. hirtaHerbal (GATAS-GATAS) (Makahiya)(MAKAHIYA) Plant Extract +1 +1 +1 Total3 Mean1 P. rubra (KALACHUCHI) 12 24 43 M. pudica +2 +2 +2 (T)6 (x̅2 ) (KANGKONG) Trial 1 Trial 2 Trial 3 1.33 I. aquatica 110 132 321 C. aromaticus(Gatas-Gatas) +1 +1 +1 3 1 7 14.33 E. hirta (GATAS-GATAS) (MAKAHIYA) +1 +1 +1 3 1 M. pudica 26 83 (KANGKONG) 79 107 P.Control rubra group(KALACHUCHI)(Oregano) +1+2 +2 +1+2 46 2 (OREGANO) E. hirta (GATAS-GATAS) 246 181 C. aromaticus +1 +1 +1 3 1 (MAKAHIYA) 129 94 249 I. aquatica Grand Total +1 +2 +1 264 1.33 C. aromaticus 228 163 Control group (OREGANO) +2 +2 +2 6 2 153 580 193.33 Conrol group 112 123 384 128 M. Grandpudica Mean (KANGKONG) +2 +2 +2 6 1.441.332 (OREGANO) 298 689 229.67 Grand Total 26 Grand Total 149 2,266 E. hirtaGrand (GATAS-GATAS) Mean(MAKAHIYA) +1 +1 +1 3 1.441 Grand Mean 125.89 C. aromaticus +1 +1 +1 3 1

Control group (OREGANO) +2 +2 +2 6 2

Grand Total 26 Grand Mean 1.44 32 UV Journal of Research

Pastoril, M. F. Experiment No. 3 Table 1.2. Summary of Data Gathered on Experiment No.3 Grade Level of Clot Formation, Turbidity and Colony Count Trial 1 Trial 2 Trial 3 with the mean of 2 did not show any inhibitory on the Inhibitory Assay Against S. Grade Level aureus Coagulase Production Using Pooled Plasma (A,B,C)Grade Level Grade Level plant extracts against coagulase production activity against clot formation since it was the Herbal Plant Extract Clot Formation Colony Clot Formation Colony Clot Formation Turbidity Turbidity Turbidity The observed clot formation of the five herbal of 2 did not show any inhibitory activity against Count Count Colony of S. aureus may attribute to the compound it same as the mean of the control group based on 4 hrs 24 hrs 4 hrs 24 hrsExperiment No. 3 4 hrs 24 hrs Count plant extracts against coagulase production clot formation since it was the same as the mean Experiment No. 3 E.E. hirtahirta (GATAS-GATAS) and C. Trial 1Less Trial 2Less Trial 3 may attribute to the compound it of the control group based on Table 2.1 data. The P. rubra No clot +1 12 No clot +2 No clot +1 Less Turbid 24 (KALACHUCHI) TrialTurbid 1 TrialTurbid 2 Trial 3 aromaticus not prove that the plant extractsS. aureus inhibit only the Grade Level Grade Level Grade Level possesses. The (Gatas-Gatas) and Tableresults 2.1 for data. clot The formation results dofor not clot prove formation that thedo I. aquatica Less Less 7 greater effect in inhibiting clot formation with S. aureus coagulase production thus observation Herbal Plant Extract NoClot clot Formation +1 Colony NoClot clot Formation +2 Colony110 NoClot clot Formation +1 Less Turbid 132 Grade Level TurbidityTurbid Grade Level TurbidityTurbid Grade Level Turbidity (Oregano) (OREGANO) were were found found to have to greater have plant extracts inhibit only the coagulase Herbal Plant Extract Clot Formation ColonyCount Clot Formation ColonyCount Clot Formation Colony P. rubra P. rubra (KALACHUCHI)I. aquatica and on turbidity and colony count were performed to 4 hrs 24 hrs TurbidityLess 79 4 hrs 24 hrs TurbidityLess 4 hrs 24 hrs Turbidity Count (KANGKONG)M. pudica No clot +2 Count No clot +2 Count26 No clot +2 Less Turbid Colony effect in inhibiting clot formation with the mean production thus observation on turbidity 4 hrs 24 hrs Turbid 4 hrs 24 hrs Turbid 4 hrs 24 hrs Count I. aquatica Less Less P. rubra No clot +1 12 No clot +2 No clot +1 Less Turbid 24 ofthe 1. mean The of 1. The (Kalachuchi) and and colony count were performed to further (KALACHUCHI)E. hirta (MAKAHIYA) Turbid 129 Turbid 94 in terms of inhibiting the clot formation with P. rubra No clot +1 TurbidLess No clot +1 TurbidLess 246 No clot +1 Turbid 181 (Kangkong) showed the same effect in terms of investigate inhibitory activity of the five herbal No clot +1 Less 12 No clot +2 Less 7 No clot +1 Less Turbid 24 (KANGKONG) showed the same effect further investigate inhibitory activity of the five (KALACHUCHI)I. aquatica No clot +1 Turbid No clot +2 Turbid 110 No clot +1 Less Turbid 132 M. pudica M. pudica C. aromaticus Turbid 153 Turbid (GATAS-GATAS) No clot +1 Turbid 228 No clot +1 Turbid No clot +1 Turbid 163 inhibiting the clot formation with the mean of herbalplant extracts. plant extracts. I. aquatica Less 79 Less 7 (KANGKONG)M. pudica No clot +1+2 Less No clot +2 Less 11026 No clot +1+2 Less Turbid 132 Conrol group No clot +2 Turbid 112 No clot +2 Turbid 298123 No clot +2 Less Turbid 1.33.the Table mean While 2.2. of Turbidity of Five Different Plant Extracts and Control Group in Tube Coagulase Test Turbidity 1.33. While of(Makahiya) five different with (MAKAHIYA)plant the extracts mean and control group in Tube Coagulase Test (OREGANO) Turbid Turbid (MAKAHIYA) Less 12979 Less 94 (KANGKONG)E. hirta No clot +2 No clot +2 26 No clot +2 Less Turbid M. pudica No clot +1Turbid No clot +1Turbid 246 No clot +1 Turbid 181149 C. aromaticus 153 E.(GATAS-GATAS) hirtaThe (MAKAHIYA) results Noin clot experiment +1 Turbid 1 using129 228 Plasma No clot The +1 number Turbid of colonyNo clot growth +1 obtained Turbid shown 16394 Replication No clot +1 Turbid No clot +1 Turbid 246 No clot +1 Turbid 181 Herbal Plant Extract Less Less 298 (OREGANO)Conrol group No clot +2 112 No clot +2 123 No clot +2 Less Turbid C. aromaticus Turbid 153 Turbid (GATAS-GATAS) No clot +1 Turbid 228 No clot +1 Turbid No clot +1 Turbid 163 Trial 1 Trial 2 Trial 3 plasma used in experiment 3 was pooled plasma149 Less Less P. rubra (KALACHUCHI) Less Turbid Less Turbid Less Turbid XConrol and group experimentNo clot 2 using +2 Plasma Y112 in grade No clot on +2 Table 1.2 was298123 less No clotthan 300 +2 cfu/ml. Less Turbid Also, the grade(OREGANO)The level results and inturbidity experiment wereTurbid 1measured using Plasma after The numberTurbid of colony growth obtained shown level after 4 hours, no clot were formed. The I. aquatica Less Turbid Less Turbid Less Turbid 149 (Kalachuchi) levelfrom A,of B clot and C.formation of experimental group M. pudica Less Turbid Less Turbid Less Turbid The results in experiment 1 using Plasma Theplasma number used inof experimentcolony growth 3 was obtained pooled plasmashown (Kangkong)(KANGKONG) X24 andhours. experiment 10µl of the 2 mixture using Plasma in each Y tube in gradewere on TableThe first 1.2 partwas dealsless than with 300 measuring cfu/ml. theAlso, grade the E. hirta (GATAS-GATAS) Turbid Turbid Turbid grade level and turbidity were measured after and control group 24 hours of observation and (Makahiya)(MAKAHIYA) levelobtained after and 4 immediately hours, no clot streaked were in formed. MHA with The of incubation, numerous growths of colonies counting the number of colonies formed in MHA C. aromaticus(Gatas-Gatas) Turbid Turbid Turbid the use of bacterial cell spreader. After 24 hours forlevelfromplasma in A,of each Bused clot and testin C.formation experiment solution of including 3experimental was pooled the controlplasmagroup Conrol group Less Turbid Less Turbid Less Turbid X and experiment 2 using Plasma Y in grade on Table 1.2 was less than 300 cfu/ml. Also, the (Oregano)(OREGANO) grade24 hours. level 10µl and of turbidity the mixture were in measuredeach tube wereafter and Thecontrol first grouppart deals 24 hours with measuring of observation the grade and ofwereobtainedlevel incubation, seen. after and Thus, 4 immediately hours, numerous the amount no clot streakedgrowths of were mixture in formed.of MHA streakedcolonies with The counting the number of colonies formed in MHA The P. P.rubra rubra (KALACHUCHI), I.I. aquatica that the coagulase production of S. aureus was the use of bacterial cell spreader. After 24 hours inTable MHA 2.1. wasGrade Level of Clot Formation of Five Different Plant Extracts and Control Group in Tube Coagulase Test reduced to 0.1µl in experiment 3. levelforgroupfrom in A, ofandeach B clot and also test C.formation observing solution ofits including turbidity.experimental the controlgroup M. M. pudica pudica 24 hours. 10µl of the mixture in each tube were The first part deals with measuring the grade groupThe showed the same(Kalachuchi), characteristic in terms of that the coagulase production of was and control group 24 hours of observation and (Kangkong) (Makahiya) and Control inhibited or the bacteria were killed by the obtainedwere seen. and Thus, immediately the amount streaked of mixture in MHA streaked with turbidity(KANGKONG) which is less turbid (MAKAHIYA) compared and to ControlE. hirtaE. designatesinhibited orthe the presence bacteria of more were bacteria killed by in the of incubation, numerous growths of coloniesReplication counting the number of colonies formed in MHA inTable MHA 2.1. wasGrade Level of Clot Formation of Five Different Plant Extracts and Control Group in Tube Coagulase Test reduced to 0.1µl in experiment 3. group and also observingTreatment its turbidity. Treatment grouphirta(GATAS-GATAS) showed theand same C. C.aromaticus characteristic aromaticus in terms solutionpresence and of that the the herbal bacteria plant did extracts. not form Turbid a clot the use Herbalof bacterial Plant cellExtract spreader. After 24 hours for in each test solutionTotal including theMean control of turbidity which is less turbid compared to designates the presence of more bacteria in the (T) (x̅ ) Trial 1 Trial 2 Trial 3 (Gatas-Gatas) and (OREGANO)(Oregano) solution and that the bacteria did not form a clot Replication were seen. Thus, the amount of mixture streaked Treatment Treatment showed in Table 2.2. Less turbid either indicates thus making the solution turbid. P. rubraHerbal (KALACHUCHI) Plant Extract +1 +2 +1 Total4 Mean Table 2.3. ColonyColony Count (CFU/ml) of Five Different Plant Extracts and Control Group in Tube Coagulase Test Count (CFU/ml) of five different plant extracts and control group inTube Coagulase Test inTable MHA 2.1. wasGrade Level of Clot Formation of Five Different Plant Extracts and Control Group in Tube Coagulase Test reduced to 0.1µl in experimentTrial 1 3. Trialgroup 2 and Trial also 3 observing(T) its turbidity. (x̅ ) I. aquatica +1 +2 +1 4 1.33 Replication Treatment Treatment P. rubra (KALACHUCHI) +1 +2 +1 4 Herbal Plant Extract Total Mean M. pudica (KANGKONG) +2 +2 +2 6 1.332 (T) (x̅ ) Replication Treatment Treatment Trial 1 Trial 2 Trial 3 I. aquatica +1 +2 +1 4 1.33 E. hirtaHerbal (GATAS-GATAS) (MAKAHIYA) Plant Extract +1 +1 +1 Total3 Mean1 P. rubra (KALACHUCHI) 12 24 43 M. pudica +2 +2 +2 (T)6 (x̅2 ) (KANGKONG) Trial 1 Trial 2 Trial 3 1.33 I. aquaticaaquatica 110 132 321 C. aromaticus +1 +1 +1 3 1 (Kalachuchi) 7 14.33 E. hirta (GATAS-GATAS) (MAKAHIYA) +1 +1 +1 3 1 M. pudica 26 83 (Kangkong)(KANGKONG) 79 107 P.Control rubra group(KALACHUCHI) +1+2 +2 +1+2 46 2 (OREGANO) E. hirta (GATAS-GATAS) 246 181 C. aromaticus +1 +1 +1 3 1 (Makahiya)(MAKAHIYA) 129 94 249 I. aquatica Grand Total +1 +2 +1 264 1.33 C. aromaticus 228 163 Control group (OREGANO) +2 +2 +2 6 2 (Gatas-Gatas) 153 580 193.33 Conrol group 112 123 384 128 M. Grandpudica Mean (KANGKONG) +2 +2 +2 6 1.441.332 (Oregano)(OREGANO) 298 689 229.67 Grand Total 26 Grand Total 149 2,266 E. hirtaGrand (GATAS-GATAS) Mean(MAKAHIYA) +1 +1 +1 3 1.441 Grand Mean 125.89 C. aromaticus +1 +1 +1 3 1

Control group (OREGANO) +2 +2 +2 6 2

Grand Total 26 Grand Mean 1.44 34 UV Journal of Research 35

Pastoril, M. F.

Any two means having a common letter largest treatment mean which is the Control group However, the inhibitory activity against The second part deals with determining of 128 and the third Rlargest R valueR (the R value S. aureus coagulase production was further treatment mean whichp is the controlp p group pof 128 determined by the number of colony growth in and colony count of the control group and are not significantly different at the 5% level of and the third largest value (the value at p = 4) the significant mean difference of grade level meanP. rubra of P. rubra experimental group using one-way ANOVA and significance. atof 81.53p=4) of is 12881.53 - 81.53 is 128-81.53= = 46.47. Because 46.47. Becausethe mean the of The above table (Table 3.3) showed that there (Kalachuchi) (KALACHUCHI) is less than 46.47, is less it than is declared 46.47, componentsMHA. The variation that plant of resultsextracts in possesses the colony against count is no significant difference between Makahiya and largestitsignificantly is declared treatment differentsignificantly mean from differentwhich theI. aquatica mean is from the of thetheI. aquatica mean control of S.based aureus on Table 2.3, may due to the different DMRT. formationControl group; of Kalachuchi and Kangkong; Gatas- thegroup. Control The group. difference The difference between between the fourth the largest fourth value S. aureu Rp Rp Gatas and Oregano in terms of inhibiting the clot treatment(the mean which is the (Kangkong) Table 3.1.. Analysis of Variance (ANOVA) of Grade Level of Clot Formation of Five Different Plant Extracts and Rp s in Tube Coagulase Test. (KANGKONG) of 107 and the fourth largest Control Group in Tube Coagulase Test TableTable 3.4.3.4. Duncan’sDuncan’s Multiple Multiple Range Range Test Test (DMRT) (DMRT) for Becauseof 107 and theP. rubrathe mean fourth of P. largest rubra (KALACHUCHI)value (the isvalue less Colonyfor Colony Count Count of five of Five different Different plant Plant extracts Extracts and control and at p = 3) value of 79.08 at p=3) is 107-79.08 of 79.08 =is 27.92 107-79.08=27.92 Because the Degree of Sum of Mean Tabular F group in Tube Coagulase Test using the Alphabet mean of (Kalachuchi) is less than 79.08, it Source of Variation Computed F Control Group in Tube Coagulase Test Using the Alphabet the meanI. aquatica of the I. aquatica Freedom Square Square 5% 1% Notation Notation becausethanis declared 79.08, theP. significantlyitrubra mean is declared of P. rubra differentsignificantly (KALACHUCHI) from different the mean is from the of Treatment onlythe outside the(Kangkong). groupings (KANGKONG). already HoweverP. rubra made, because However,However P. rubra the Experimental Error 12 Mean Herbal Plant Extract DMRT mean of (Kalachuchi) is the only outside 5 3.11 0.62 5.64** 3.11 5.06 (x̅) (KALACHUCHI) mean was compared using theR Total the groupings already made, (Kalachuchi)p 1.33 0.11 appropriate Rp values with the rest of the means C. aromaticusaromaticus a mean was compared using the appropriate The computed F value is larger17 than the tabular 4.44 F namely ControlI. aquatica group, I. aquatica M. pudica valuesM. pudica with the rest of the means namely control E. hirtahirta (GATAS-GATAS) (OREGANO)(Oregano) 299.67 b value at the 5% and 1% level of significance. This group,the only one whose(Kangkong) difference (KANGKONG) is and less than andthe F value in Rp Conrol group(Gatas-Gatas) 193.33128 c (Makahiya). (MAKAHIYA). Of the three Ofcomparisons, the three comparisons, the only one implies that the treatment difference in grade level of clot formation of five different plant extracts and corresponding Rp valueM. is pudica that between M. pudicaP. control group is said to be highly significant indicated by two asterisks (**) on the computed rubrawhose difference is P.less rubra than the(KALACHUCHI) correspondingR 83- I.Control aquaticaaquatica group d p the analysis of variance. value is that betweenR M. pudica (Makahiya)M.P. pudica andrubra Table 3.2. Analysis of Variance (ANOVA) of Colony Count of Five Different Plant Extracts and Control Group in (MAKAHIYA)(Kalachuchi) andp 83 - 14.33 = 68.67 < (at p = Tube Coagulase Test M. pudica (KANGKONG)(Kangkong) 10783 e P. rubra (KALACHUCHI) are M. pudica 2)14.33=68.67 of 75.41. Thus < (at p=2) of(Makahiya) 75.41. Thus and Degree of Sum of Mean Tabular F Source of Variation Computed F P. rubrarubra (KALACHUCHI) (MAKAHIYA)(Makahiya) (MAKAHIYA)(Kalachuchi) are and declared not significantly different Freedom Square Square 5% 1% declaredfrom each not other. significantly different from each other. (Kalachuchi) 14.33 Treatment The above above table table (Table (Table 3.4) 3.4)showed showed that there that Experimental Error 12 21,448 Any two means having a common alphabet thereis no is significant no significant difference difference between between Oregano Oregano and Total 5111,339.78 89,891.78 17,978 10.06** 3.11 5.06 Any two means having a common alphabet 1,787.33 andGatas-Gatas; Gatas-Gatas; Gatas-Gatas Gatas-Gatas and control and Control group; control group; The computed value is larger than the tabular are not significantly different at the 5% R level of F 17 F are not significantly different at the 5% levelp of Controlgroup, KangkongGroup, Kangkong and Makahiya; and Makahiya; Kangkong Kangkong and significance.The difference between the largest Rp value (The Rp andMakahiya; Makahiya; Makahiya Makahiya and and Kalachuchi Kalachuchi in in terms terms of value at the 5% and 1% level of significance, significance.R The difference between the largest value F p C. aromaticus Colony Count. mean(The C. aromaticusvalue at p = 6) of 83.34 and the largest herbal plants that have inhibitory effect against S. the treatment difference in colony count of five different plant extracts and control group is said to be The last part deals with the identification of highly significant indicated by two asterisks (**) on the computed value in the analysis of variance. treatment value at p=6) mean of 83.34 and the largest(Oregano) treatment of aureus Table 3.3. Duncan’s Multiple Range Test (DMRT) for Comparing Grade Level of Clot Formation of 299.67all treatments is 299.67 means, (OREGANO) - except 83.34 that = of 146.33. of 299.67 E. hirta Fromis (GATAS-299.67- the E. hirta (GATASGATAS), coagulase production. Five Different Plant Extracts and Control Group in Tube Coagulase Test Using the Alphabet Notation arrayGATAS),83.34= of 146.33. means From obtained, the array all treatmentsof means obtained, means, TableTable 4. Identification Identification of of Herbal herbal Plants plants That that Have have except that of are less than InhibitoryInhibitory Activity Activity Against against Coagulase Coagulase Production Production of of S. S. Mean C. aromaticus Herbal Plant Extract DMRT are less than computed difference of 146.33. aureusaureus (x̅) computedfrom C. aromaticus difference of 146.33. Hence, they are Hence, they are declared significantly different declaredbetween the significantly second largest different treatment from mean E. hirta Anti- Anti- M. pudica 2 a b E. hirta of Herbal Plant Extract (Oregano).(GATAS-GATAS) The difference (OREGANO). and between the Thesecond the difference secondlargest bacterial coagulase Rp Rp Conrol group (MAKAHIYA) 2 largestR value treatment (the R mean (Gatas-Gatas) P. rubra (KALACHUCHI) + p pof 193.33 P. rubra (KALACHUCHI) c d 193.33 and the second largest value (the I. aquatica(Kalachuchi) + value at p=5) of 82.35 is 193.33- M. pudica + I. aquatica valueall treatments at p = 5) means,of 82.35 except is 193.33 that -of 82.35 Control = 110.97. group (Kangkong)(KANGKONG) 1.33 82.35=110.97. From the array of means obtained, From the array of means obtained, all treatments E. hirta (Makahiya)(MAKAHIYA) - + E. hirta (GATAS-G (KANGKONG)ATAS) 1.331 e f means, except that of control groupE. arehirta lessE. hirta than C. aromaticus(Gatas-Gatas)(GATAS-GATAS) - + C. aromaticus 1 are less than computed difference of 110.97. Thus computed(GATAS-GATAS).. difference The difference of 110.97. between Thus the they third are (OREGANO) they are declared significantly different from (Oregano) (OREGANO) declared significantly different from (Gatas- Gatas) The difference between the third largest UV Journal of Research 2014 36 UV Journal of Research 37 Inhibitory activity of Sandoricum koetjape Merr. (santol) In this study, the inhibitory activities of REFERENCES the herbal plant extracts against coagulase Calmorin, L.P., & Calmorin, M.A. (2010). Research Sergio Jabel II productionIn the in study, S. aureus the inhibitorywere assessed activities through of nd methods and thesis writing (2 ed.). Manila: Rex Maria Feibe Pastoril the herbal plant extracts against coagulase Bookstore Inc. leaf extract to Blood Type O fibrin formationJosephine Fe Genest, S., Kerr, C., Shah, A., Rahman M., Saif-E-Naser, productionresulting clot in in each tubewere was assessed measured through and Sheryll Sacramento Tube Coagulase Test and MHA colony count. The G., Nigam, P., Nahar, L., & Sarker, S. (2008). Comparative bioactivity studies on two Mimosa College of Pharmacy resultingresults for clot the inTube each Coagulase tube was Test measured and colony and species. Boletín Latinoamericano y del Caribe de University of the Visayas the growth of colonies in MHA was counted. The thecount growth revealed of colonies that only in the MHA plant was extract counted. of E. hirta TheE. Plantas Medicinales y Aromáticas, 7(1), 39-43. results for the Tube CoagulaseC. aromaticus Test and colonyJ Joshi, B., Lekhak, S., & Sharma, A. (2009). Antibacterial hirta C. aromaticus property of different medicinal plants: [email protected] countshowed revealed inhibitory that activity only theagainst plant the extract production of Ocimum sanctum, Cinnamomum zeylanicum, Date submitted: April 11, 2014 Date of acceptance: June 30, 2014 (GATAS-GATAS) and (OREGANO) of coagulase(Gatas-Gatas) in S. aureus and (Oregano)P. rubra Xanthoxylum armatum and Origanum majorana. Kathmandu University Journal of Science, showed(KALACHUCHI) inhibitory and activity I. I.aquatica againstaquatica the production . Furthermore, the Engineering and Technology, 5(1), 143–150. ABSTRACT ofplant coagulase extracts in were effective. Furthermore, in inhibiting the theK Kardono, L., Tsauri, S., Padmawinata, K., Pezzuto, J., & coagulase(Kalachuchi) production and due to its (KANGKONG)antibacterial(Kangkong) Kinghorn, A. (1990). Cytotoxic constituents This study was conducted to evaluate the inhibitory activity of Sandoricum plantactivity extracts against werethe bacteria effective S. aureus in inhibiting the of the bark of Plumeria rubra collected in S. aureus Indonesia. Journal of Natural Products, 53(1), coagulasethe plat extract production of M.M. pudica pudica due to its antibacterial 1447-1455. . Meanwhile activityshow any against inhibitory the bacteria activity against. Meanwhile, S. aureusL Larsen, H.S., Mahon, C.R., & Manuselis, G. (2006). koetjape Merr. leaf extract to blood type O fibrin formation, where warfarin the plat extract of (MAKAHIYA)(Makahiya) diddid not Diagnostic microbiology (3rd ed). Saunders. M Majumdar, P., Bhakta J., & Munekage, Y. (2009). (positive), negative control and different extract concentrations (50%, 75% and show any inhibitory activity against 100%) were compared. The computed F-value for the treatment was 238 with coagulase production. Antimicrobial efficacies of methanol extract VIII.coagulase CONCLUSION production. of Asteracantha longifolia, Ipomoea aquatica degrees of freedom 2.7. For replication, the computed F-value obtained was 3 The results obtained from the measurement and Enhydra fluctuans against Escherichia concentrationwith degrees ofhas freedom the highest 2.7. Thetime formerof coagulation F-value butwas replication significant was at 0.5 almost and the0.1 of grade level, turbidity and colony count showed coli, Pseudomonas aeruginosa, Staphylococcus aureus and Micrococcus luteus. The Internet level of probability and the latter, was not significant. Results showed that 100% that TheE.E. hirta hirtaresults obtained from the C.C.measurement aromaticus Journal of Alternative Medicine, 7(2), 1540- of grade level, turbidity and colony count showed 2584. Keywords:same. inhibitory activity; fibrin formation; coagulation; anticoagulant (GATAS-GATAS) and N Nostro, A., Roccaro, A., Bisignano, G., Marino, thatactivity against(Gatas-Gatas) the production and of coagulaseS. aureus in (OREGANO) plant extracts have inhibitory A.,Cannatelli, M., Pizzimenti, F., Cioni, P., (Oregano)S. aureus plant extracts have inhibitory activity Procopio, F., & Blanco, A. (2007). Effects of I. INTRODUCTION Warfarin and heparin and also an antiplatelet againstto the synergisticthe production effects of coagulaseof its constituents in or. oregano, carvacrol and thymol on Staphylococcus Coagulation is the process by agent such as aspirin are used in disorders of Thethe presence inhibitory. The of inhibitory activity bioactive might activity compounds be might due be toof due the aureus and Staphylococcus epidermidis biofilms. Journal of Medical Microbiology, 56(1), 519– synergistic effects of its constituents or the 523. which blood forms clots (Finkel, Clark & coagulation (Zehnder, 2012; Martin, 2009; ). presencestudy do not of prove bioactive that compoundsplant extractsE. hirta of of the E. hirta twoO Ogueke, C., Ogbulie, J., Okoli, I., & Anyanwu, B. (2007). of haemostasis, the cessation of blood loss from of unhealthy blood vessels clogged with a build two herbal plant extracts. The results of this herbal plant extracts. C.The aromaticus results of the study do Antibacterial activities and toxicological aCubeddu, damaged 2009).vessel, Itwherein is an a importantdamaged blood part Blood-clot strokes can also happen as the result C. aromaticus potentials of crude ethanolic extracts of not prove that plant extracts of (Gatas- Euphorbia hirta. Journal of American Science, vessel wall is covered by a platelet and fibrin- body regards these buildups as multiple, tiny (GATAS-GATAS) and (OREGANO) Gatas)conducted and do not, in any(Oregano) way, stimulate already hasthe 3(3), 11-16. containing clot to stop bleeding and begin repair up of fatty deposits and cholesterol. The human already has therapeutic value. The in vitro tests therapeutic value. The in vitro tests conductedSa Saeed, S., & Tariq, P. (2005). Antibacterial activities of So the body reacts to these injuries just as it Mentha piperita, Pisum sativum and Momordica and repeated injuries to the blood vessel wall. doresults not,E. warrant hirta in any way,the E. stimulate hirta the C. complexity aromaticus ofC. would if you were bleeding from a wound; it complexity of the human body. Instead, these charantia. Pakistan Journal of Botany, 37(4), thearomaticus human body. Instead, these results warrant 997-1001. of the damaged vessel. Disorders of coagulation (GATAS-GATAS) and the (Gatas-Gatas) and Stoppler, M.C. (2009). Staphylococcus infection article. instantlycan lead after to an an increased injury to the risk blood of bleeding vessel has or According to the 2012 morbidity and (Oregano) plant(OREGANO) extracts toplant further extracts anti-coagulase to further Disease and Condition. Available from: http:// obstructive clotting. Coagulation begins almost responds by forming clots. www.medicinenet.com/staph_infection/article.htm investigation.anti-coagulase investigation. originality index: 91 % Todar, K. (2004). Todar’s Online Textbook of Bacteriology: Cardiovascular disorders such as hypertension The Good, the bad, and the deadly. SCIENCE damaged the endothelium lining of the vessel. mortality chart book on cardiovascular, lung, similarity index: 9 % Magazine, 304:1421. including arteriosclerosis and congestive and blood diseases, stroke ranked seventh for paper id: 470239059 heartand thrombosisfailure which are is wellcaused known by problem in adults in diseasethose people was the aging third from leading 45-64 cause years of and death fourth for Grammarly: checked blood circulatory system as blood clotting on those aged 65 years and older in 2008. Heart

those aged 25-44 years, second for those aged disorders constitute a serious medical problem. 45-64 years, and first for those aged 65 years