1

Chapter 1

THE PROBLEM AND ITS BACKGROUND Introduct on

A hand sanitizer commercially available or naturally prepared is a supplement or alternative to hand washing with soap and water. Various preparations are available, including gel, foam, and liquid solutions. The active ingredient in hand sanitizers may

be isopropanol, ethanol, n-propanol, or povidone-iodine. Inactive ingredients in alcohol rubs typically include a thic ening agent such as polyacrylic acid for alcohol gels, humectants such as glycerin for liquid rubs, propylene, and essential oils of plants. Alcohol based hand sanitizers are more effective at illing germs than soaps and do not dry out hands as much. !onsumer alcohol-based hand sanitizers, and health care hand alcohol or alcohol hand antiseptic agents, are antiseptic products used to avoid transmission of pathogens.

It is well- nown that the primary vector of transmission for disease causing pathogens is hand contact. "ven when rigorous protocols for hand washing and hygiene are followed, pathogens present on any contact surface can easily recolonize recently sanitized hands. #ealth care settings in particular increasingly demonstrate the problems of pathogen transmission

leading to nosocomial infections due to the number of the highly susceptible populations in health care facilities. $ther problems associated with regulatory pressures are also demonstrated by the fact that %edicare is planning to no longer reimburse for hospital-acquired infections& particularly %ethicillin 'esistant (taphylococcus aureus )%'(A*.

The !enter for +isease !ontrol suggests that there are two main ways to contract any of the common viral illnesses. !ontracting may occur through coughing or sneezing with airborne illness ) nown as ,droplet spread-*, or through direct contact with germs spread through touching of contaminated items. !oughing and sneezing of infected individuals propel respiratory droplets of infection through the air and deposit on the mouth or nose of people in the surrounding vicinity, spreading the ailment often without notice. $ftentimes, influenza germs may remain on ob.ect surfaces affected by infected individuals, leaving those surrounding persons vulnerable to the second most common form of disease contraction, direct contact. +irect contact may occur in two ways, including through contact with an infected individual through touching and sharing of personal items such as food or silverware, or through surface contact, meaning that an uninfected individual becomes e/posed to the harmful bacteria left behind on ob.ects touched, coughed on, or sneezed on by infected individuals post interaction with the ob.ect or surface. %ost often, individuals ma e contact with surfaces tainted with

respiratory droplets, and touch the eyes or mouth before properly disinfecting, resulting in the spread of disease. $nce infected, presenting symptoms of viral and respiratory disease include coughing, sneezing, fever, fatigue, muscle or body aches, wea ness, headaches, and sore throat. 0ith today1s increased rates of flu and sic ness, government officials push for the continual use of preventative techniques employed to reduce the spread of disease.

The !enters for +isease !ontrol says the most important way to prevent the transmission of dangerous diseases is to frequently wash your hands with soap and water and2or use a hand sanitizer. It also has high viricidal activity against many different inds of viruses,

including enveloped viruses such as the flu virus, the common cold virus, and #IV, though is notably ineffective against the rabies virus. Alcohol rub sanitizers are not very effective against 3orovirus )winter vomiting virus* unless they are combined with benzal onium chloride in a hand sanitizer. Alcohol rubs also ill fungi. 4niversity of Virginia %edical (chool

researchers concluded that hand sanitizing is more effective against fighting the common cold than hand washing. Alcohol ills both pathogenic )disease causing* microorganisms as well as resident bacterial flora, which generally do not cause illness. 'esearch shows that alcohol hand sanitizers do not pose any ris by eliminating 5good5 germs that are naturally present on the s in.

The body quic ly replenishes the good germs on the hands, often ma e moving them in from .ust up the arms where there are fewer harmful germs.

Antibacterial hand sanitizers either commercial or natural are mar eted to the public as an effective way to 5wash one6s hands5 when traditional soap and water are not available. These 5waterless5 products are particularly popular with parents of small children. %anufacturers of hand sanitizers claim that the sanitizers ill 77.7 percent of germs. #and sanitizers commercially available or made of natural plant e/tracts wor by stripping away the outer layer of oil on the s in. This usually prevents bacteria present in the body from coming to the surface of the hand. #owever, these bacteria that are normally present in the body are generally not the inds of bacteria that will us sic . Antibacterial agents are of big help in the society by preventing the growth of bacteria. (ince commercialized antibacterial agents are costly, people tend to find alternative ways of preventing this diseases by utilizing natural and effective and yet cheaper antibacterial agents. !ommon non-alcohol, rinse-free hand sanitizer brands use either small concentrations of the nitrogenous cationic surface-acting agent benzal onium chloride or the chlorinated aromatic compound triclosan or povidone-iodine. Alcohol free hand sanitizers may be effective immediately while on the s in, but the solution themselves can become contaminated because alcohol is an in-solution preservative and without it, the alcohol free solution itself is susceptible to contamination. #owever, even alcohol containing hand sanitizers can become contaminated if

the alcohol content is not properly controlled or the sanitizer is grossly contaminated with microorganisms during manufacture. 8ross contamination of any hand sanitizers by bacteria during manufacture will result in the failure of the effectiveness of that sanitizers and possible infection of the treatment site with the contaminating organism. (ome brands started combining alcohol rubs with natural products and essential oils to provide lu/ury lines for the growing s in care industry. These lu/ury products contain 9:; alcohol necessary to ill germs effectively and safely, but employ additional ingredients to moisturize and condition the s in.

The effectiveness of hand sanitizers depends on the number of issues, including the alcohol content of the product and how the product is used. #and sanitizers are not meant as a constant substitute for hand washing< however, these hand sanitizers can be effective supplement to the traditional hand cleaning, particularly when soap and water are not available. The (wine flu of =::7 has conducted to hand sanitizers1 squirts on the hand across the globe. >eople have been seen using hand sanitizers at public places, before eating, after using restrooms, at wor places, etc.

Therefore using hand sanitizers is very important in avoiding some bacteria being acquired in all our activities involving the use of the hands such as after using the restroom, after wor ing and before eating especially when there is no available water or maybe soaps. This study is a great help to all health wor ers and to all individuals especially children who are fun of

soils and dirty things. It will give more nowledge to the health wor ers on how to minimize the spread of bacteria and give also tips for them to prevent the e/isting bacteria in the hospitals.

Conceptua! "ra#e$or% The study of the comparative analysis on the effectiveness of hand sanitizers and !alamansi fruit and Talisay leaves e/tract against ". coli, >. aeruginosa, (. aureus was based on the study of Ahmed, %.8. et. al. entitled ,Antimicrobial Activity of ?eaf and @lower "/tract of (pathodea !ampanulata >. Aeauv. The theory involved in their study was more on the antimicrobial activity of the leaf and flower e/tract of (pathodea !ampanulata >. Aeauv against specific bacteria which is the same with the researchers1 study. Another theory was based on the .ournal of Agriculture, @ood and Analytical Aacteriology entitled ,Antibacterial Activity of >lant "/tracts on @oodbourne Aacterial >athogens and @ood (poilage Aacteria-. This theory states that a certain e/tract has a strong antimicrobial activity against a certain test organism. "ach e/tract has a different antimicrobial effect against certain organism as implicated by their zone of inhibition. The researchers modified the procedures based on the two studies by limiting their test organism into three and by using different plant e/tracts and by also utilizing hand sanitizers.

Re&earch Parad '#

E(per #enta! Group (afeguard 8reen !ross #ygieni/ Talisay !alamansi

Or'an &# Te&ted

Contro! Group)*ar a+!e

". coli >. aeruginosa (. aureus

B:; "thyl alcohol

E,,ect -ene&& &ho$n B. the /one O, Inh + t on

State#ent o, the Pro+!e# The main ob.ectives of this study are to determine the potential of natural plant e/tracts as antibacterial agents compared to the commercialized hand sanitizers. (pecifically, it aims to answer the following questionsC D.* +etermine the difference in the zone of inhibition in terms ofC a. types of bacteria used b. disinfectant used =.* +etermine the interaction effects in the zone of inhibition between the types of bacteria

and disinfectant used.

Nu!! H.pothe& &

There is no significant difference on the effectiveness of the commercialized hand sanitizers and the natural plant e/tracts.

Scope and De! # tat on

The researchers focused only their study on the effectiveness of Talisay leaves )Terminalia catappa* and !alamansi )Citrus microcarpa bunge) fruit e/tracts compared to commercialized hand sanitizers to the selected bacteria which includes Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. @urthermore the researchers limited their wor on testing the effectiveness of the two natural plant e/tracts and so with the different products of hand sanitizers to the cultured bacteria. The Talisay leaves were gathered at the compound of the !agayan (tate 4niversity, Andrews1 campus. The !alamansi on the other hand were bought at the +on +omingo >ublic %ar et and the three hand sanitizers were bought at the drug store while the test organism were purchased at +$(T laboratory, (an 8abriel, Tuguegarao !ity. The e/periment was conducted at the +epartment of (cience and Technology ?aboratory Testing !enter. The study covered the months of Eune-$ctober =:DF.

S 'n , cance o, the Stud.

3owadays, infectious diseases are the most worldwide problem. (ometimes, we are not aware that we acquired many infectious diseases because of improper hygiene specially our hand. This research is e/pected to benefit the followingC

Con&u#er&)Hand San t 0er U&er&1 The findings of the of this study will be beneficial to all consumers and users of hand sanitizers for they will be given ideas on the benefits of using hand sanitizers and they will be able now that hand sanitizers are also the best hand hygiene alternatives if soap and water are not available.

Student& 2 Pro,e&& on& o, Med ca! Cour&e&1 To students and professionals of medical courses, this study will be beneficial to them in such a way that it gives them some information on what are the benefits of using hand sanitizers. And will help them to prevent and minimize the spread of bacteria. The result of this study will be a great help to the medical students in particular. This study will give them the idea or it can be used as a basis for other researches involving hand sanitizers and also hand hygiene. This will give them a hint that hand sanitizers which will be utilize in this study will be use to prevent bacteria that they will acquire in day to day activities or wor in the hospitals. Depart#ent o, Sc ence and Techno!o'.1 Through this study, it will help the department to conduct more relevant studies on the essentials and benefits of taro plant Depart#ent o, A'r cu!ture1 This study provides an insight in the importance and benefits of medicinal herbal plants which serve as a basis for the department to promote and encourage people to plant medicinal herbal plant. Depart#ent o, Hea!th1 The finding of this study is helpful in the alternative cure of bacterial infections. They can also ma e further study for the improvement of this sub.ect matter. And

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finally, they will be able to promote the use of medicinal herbal plants in the society as an alternative treatment of some diseases.

"uture Re&earcher&1 The result of this study will give an idea to the researchers to further e/plore or investigate on the benefits and some other studies related to hand sanitizers. This will provide as a guide for the researchers to e/periment on the effectiveness of the hand sanitizers in reducing or eliminating bacteria present in our hands.

De, n t on o, Ter#& The researchers in this study used the following terms serving as conceptual and conte/tual references or au/iliary framewor of the research. A'ar1According to +elost, it is a medium in a gel a semi solid form. Bac !! 1 According to +elost, it is a rod-shape bacterium. Cocc 1 According to +elost round in shape and may appear as diplococcic )pairs*, chains, or clusters. Staphylococcus aureus. According to %c>herson and >incus, et.al, it is a gram positive cocci bacterium that is present among indigenous flora of the s in, eye, and upper respiratory tract. And it is the only staphylococcal species pathogenic to humans that produces coagulase.

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Escherichia Coli1 This is a 8ram-negative, rod-shaped bacterium that can cause serious food poisoning in humans, and are occasionally responsible for product recalls due to food contamination. Inter#ed ate1>revents small,technical factors from from causing ma.or discrepancies in the interpretation. Pseudomonas aeruginosa. According to %c>herson and >incus, it is an ubiquitous in the hospital environment and may produce serious infection in patients with burns and traumatic and operative wounds. Hu#ectant&. A substance that is used for moistening the s in. K r+.3Bauer Te&t1 According to +elost, this is used to determine the susceptibility or resistant of a pathogenic organism to a various antimicrobial agents. Moderate!. &u&cept +!e1 According to +elost , it implies strain that may be inhibited by attainable concentration of certain sanitizer. Na&oco# a! n,ect on1 According to +elost,it is an infection acquired in the hospital or other health care setting. Nor#a! ,!ora1 According to +elost, they are microorganisms normally residing in a particular body site?< they do not generally cause infeetion. Noro- ru&1 A virus that causes the ,flu,5 or vomiting and diarrhea, in people.

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Pande# c. A widespread epidemic that affects populations on different continents and that possibly circulates around the world. Patho'en1 This is an agent of disease and a disease producer. The term pathogen most commonly is used to refer to infectious organisms. These include as Staphylococcus aereus*, viruses )such as #IV*, and fungi )such as yeast*. Re& &tant1 According to +elost, it implies strain is not inhibited by the usually available systemic concentration of the agent with normal dosage. San t 0er&1 A substance or preparation for illing germs designed for use especially on foodprocessing equipment. Su&cept + ! t.1 Implies infection due to strain may be may be appropriately treated with dosage of antimicrobial agent recommended for that type of infection, unless otherwise contraindicated. S$ ne "!u1 This refers to a subset of $rthomy/oviridae that creates influenza in pigs and are endemic in pigs. This is not a phylogenetics based ta/onomiccategory. Tr c!o&an1 A pesticide agent that ill bacteria in such a way that the remaining bacteria become resistant to it. bacteria )such

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Chapter 4

RE*IE5 O" THE RELATED LITERATURE AND STUDIES

#and sanitizers are the best alternatives for hand hygiene if soap and water are not available. A great number of studies, researches and write-ups has been conducted for a period of time and still emerged as one of the top priority researches due to rapid changing and the need of

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time. This chapter is the presentation of the literature and studies from foreign and local studies and literature. E,,ect -ene&& o, hand &an t 0er& a& ant # cro+ a! Though hesitation surrounds the general public opinion regarding the effectiveness of such waterless methods, results of a recent study conducted by >ic ering, Aoehm, %wan.ali, G +avis )=:D:* suggest that waterless sanitizers are not only as effective, but in the case of eliminating harmful levels of ". coli and streptococci )especially fecal streptococci* from the body, such waterless sanitizers prevailed against washing with soap and water in geographic regions with limited supplies of soap and high incidence of infectious disease. "ffects of this research alone suggest more realistic ways to eliminate bacteria in these regions that do not brea the ban , so to spea amongst citizens with already-limited resources )>ic ering et al., =:D:*. @urther efficacy studies such as that conducted by Hampf and $stermeyer )=::I* continually provide data supporting specific types shown more effective than competing brands and alcoholbased solutions. #and sanitizer with isopropyl alcohol may instantly ill up to 77.77 percent of non-spore forming bacteria in less than F: seconds. Although isopropyl alcohol is effective at eliminating various inds of bacteria and viruses, application of hand sanitizer does not ill all types of bacteria. @or e/ample, alcohol-based hand sanitizers are significantly ineffective against the norovirus and rabies virus. #owever, hand sanitizers that contain benzal onium chloride eliminate the norovirus.

15

According to a study conducted at "ast Tennessee (tate 4niversity, the concentration of isopropyl alcohol determines effectiveness. >roducts that contain a low percentage, such as J: percent, are not as effective as products that contain a higher percentage. 'esearchers suggest products contain a concentration of over B: percent isopropyl alcohol as the sanitation standard for hospitals and clinics. )'etrieved from httpC22www.livestrong.com*

I#portance o, Hand H.' ene

Infectious diseases )I+* circulating in the home and community remain a significant concern. There are several researches that the threat of I+ will increase in coming years. Two factors are largely responsible for this trendC first, the constantly changing nature and range of pathogens to which we are e/posed and, secondly, the demographic changes occurring in the community, which affect our resistance to infection. This report reviews the evidence base related to the impact of hand hygiene in reducing transmission of I+ in the home and community. To summed up the importance of hand hygiene, these areC )D* #and hygiene is a ey component of good hygiene practice in the home and community and can produce significant benefits in terms of reducing the incidence of infection, most particularly gastrointestinal infections but also respiratory tract and s in infections. )=* +econtamination of hands can be carried out either by hand washing with soap or by use of waterless hand sanitizers, which reduce contamination on hands by removal or by illing the organisms in it. The health impact of

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hand hygiene within a given community can be increased by using products and procedures, either alone or in sequence, that ma/imize the log reduction of both bacteria and viruses on hands. )F* The impact of hand hygiene in reducing I+ ris s could be increased by convincing people to apply hand hygiene procedures correctly )e.g. wash their hands correctly* and at the correct time. )J* To optimize health benefits, promotion of hand hygiene should be accompanied by hygiene education and should also involve promotion of other aspects of hygiene. )AEI!C American Eournal of Infection !ontrol*. According to ?arson, Kuiros, G ?in )=::B* the specific guidelines for hygiene amongst hospital staff have changed since inception in D7LD to today )li e most regulated guidelines are nown to do with increasing amounts of information provided by e/tended research on a given topic*. In D7LD, the only recommendations provided to healthcare employees suggested that ,hand hygiene should occur before invasive procedures or while ta ing care of susceptible patients and touching wounds< that plain soap ,unless otherwise specified- should be used or antimicrobial hand washing product utilized pre-interaction with newborns, high-ris patients, and immunocompromised patients- )?arson et al., =::B*. Aeyond these recommendations, nothing further was specified for surgical hand preparation, s in care, fingernail treatment, education and motivation, or administration measures to increasing hand hygiene )?arson et al., =::B*.

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4niversity of Virginia %edical (chool researchers concluded that hand sanitizing is more effective against fighting the common cold than hand washing. Alcohol ills both pathogenic )disease causing* microorganisms as well as resident bacterial flora, which generally do not cause illness. 'esearch shows that alcohol hand sanitizers do not pose any ris by eliminating 5good5 germs that are naturally present on the s in. The body quic ly replenishes the good germs on the hands, often moving them in from .ust up the arms where there are fewer harmful germs. Alcohol also strips the s in of the outer layer of oil, which may have negative effects on barrier function of the s in. #owever, washing with detergents, such as commonly used hand soaps, results in a greater barrier disruption of s in compared to alcohol solutions, suggesting an increased loss of s in lipids. )'etrieved from www.!anadianliving.com* According to a study conducted at "ast Tennessee (tate 4niversity, the concentration of isopropyl alcohol determines effectiveness. >roducts that contain a low percentage, such as J: percent, are not as effective as products that contain a higher percentage. 'esearchers suggest products contain a concentration of over B: percent isopropyl alcohol as the sanitation standard for hospitals and clinics. )'etrieved from httpC22www.livestrong.com*

Bene, t& o, Hand San t 0er

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#and sanitizers are convenient, portable, easy to use and not time consuming. #and sanitizer is an instant type of cleansing material that can be safely used by children and adults in almost any setting. It is very simple to use and requires a minimal amount of effort. 0ithout using water or a towel, a pea-sized amount of hand sanitizer can be spread between the hands and fingers and then rubbed together until dry. #and sanitizer helps to prevent the spread of germs and to eep hands clean and fresh. #and sanitizers may help people avoid many types of illness. (ome e/amples include tuberculosis, some strains of influenza and the common cold. Aecause most hand sanitizers also combat fungi and bacteria, they can also be used after handling meat or food-borne contaminated surfaces. )?earnwell.orgC #and #ygiene for #ealth !are 0or ers*

In'red ent& n Hand San t 0er that K !!& Bacter a

#and hygiene is one of the most important methods in preventing the spread of disease. #ands can be host to an array of different bacteria and viruses every time you come in contact with something. #and sanitizers are an e/cellent choice of sanitation when water and soap are not available. The 4niversity of @lorida and the !enters for +isease !ontrol report that isopropyl alcohol, ethanol and triclosan are three effective hand sanitizer ingredients.

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"thanol is also referred to as ethyl alcohol or grain alcohol and it is characterized as a colorless and flammable liquid. It has widespread use in many industrial and medical applications including hand sanitizers and disinfectants. The American (ociety for %icrobiology reports that alcohols destroy bacteria by causing membrane damage and denaturation of proteins. "thanol also prevents the spread of microbes by interfering with cell metabolism and cell division. A study published in the A%! Infectious +iseases Eournal tested the effect of ethanolbased hand sanitizers on bacteria. The results reported that ethanol was effective for hand disinfection against bacteria after : and F hours of application.

Isopropyl alcohol is also called isopropanol and is a colorless flammable liquid with a strong odor. (imilar to ethanol, isopropyl is an alcohol that is effective at destroying bacteria. Alcohols such as isopropyl, are effective against mycobacteria, viruses and fungi, according to American (ociety for %icrobiology. According to the !enters for +isease !ontrol and >revention, alcohol based hand sanitizers such as isopropyl alcohol is considered antiseptics. Antiseptics destroy harmful microorganisms that are associated with infections and not easily removed with water alone. Triclosan is another commonly used antiseptic hand sanitizer ingredient. The !+! reports that triclosan is beneficial for removing or destroying all microorganisms and flora on s in surfaces. Antiseptics such as triclosan are efficient for hand sanitizing because they disinfect

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within DI seconds. Triclosan is particularly good at destroying gram-positive bacteria, according to the American (ociety for %icrobiology. The specific mode of action remains un nown to researchers, but triclosan has shown to have antibacterial and anti-inflammatory properties. Triclosan is often combined with alcohol-based disinfectants such as ethanol and isopropyl to significantly enhance its effects.

Ta! &a. !ea-e& 6Terminalia catappa7 a& ant ,un'a!8 ant para& t c and ant # cro+ a!1

The methanolic e/tracts of leaves of Terminalia catappa showed inhibitory activity on B.subtilis and S.aureus. >hytochemical analysis yielded saponin, saponin glycosides, steroid, cardiac glycoside, tannins, volatile oils, phenols and balsam )gum*. (tudy loo ed into Terminalia catappa as an alternative to the use of chemicals and antibiotics in the aquaculture industry. 'esults showed eradication of Trichodina, fish ectoparasites, at L:: ppm. $ngoing research is being done to isolate the active ingredients in the Indian almond for fish pathogen treatment. )'etrieved from httpC22www.stuart/change.com2Talisay.html*

According to 'ay $ng, Talisay leaves contain tannin that is both anti-fungal and antibacterial. In the 4nited (tates, he said, the dried leaves are sold at about I: 4( cents per leaf. And they are imported from countries other than the >hilippines. )$ng, =:D=*

Ka!a#an& 6Citrus microcarpa7 a& ant # cro+ a! Antimicrobial properties of tropical plants against D= pathogenic bacteria isolated from aquatic organismsC A study on the antibacterial activity of 7 tropical plants against D= clinical

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and pathogenic bacterial strains including Vibrio cholera, Escherichia coli, Vibrio parahemolytics, Salmonella and Streptococcus sp. showed activity against one or more species of bacteria. Citrus microcarpa was one of the most active. )=* >aper described the potential of A. sativum and Citrus microcarpa e/tracts as alternative antimicrobial agents for local edible frog culture industry. The properties of Citrus microcarpa are aromatic, antiseptic, antiphlogistic, carminative, deodorant, refrigerant. )'etrieved from

httpC22www.stuart/change.com2Halamansi.html*

Bene, t& and U&e& o, Ca!a#an&

In food and beverages, it is being processed into ready-to-drin

.uices and .uice

concentrates to use as refreshing drin s. The .uice is primarily valued for ma ing acid beverages. (ome people use its e/tract as an ingredient to some delicacies and some sauces. (ome of them boil the sliced fruits with cranberries to ma e a start sauce. In the >hilippines, the e/tracted .uice, with the addition of gum tragacanth as an emulsifier, is pasteurized and bottled commercially. This product must be stored at low temperature to eep well. >ectin can also be recovered from the peel as a by-product of .uice production. In addition, a study conducted by +r. ?ee (eong 0ei and his team from the +epartment of @isheries and Aquaculture of the 4niversity of %alaysia on the antimicrobial properties of tropical plants against D= pathogenic bacteria isolated from aquatic organisms. In their study, 7 tropical plants including calamansi were used in the test for antimicrobial activity. It was found out that all the 7 plants tested showed antimicrobial

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activity against one or more species of the D= bacteria tested. The D= bacteria strains used were Vibrio aglinolyticus, Vibrio parahaemolyticus, Vibrio harveyi, Vibrio vulnificus, Vibrio cholera, ". coli, !itrobacter freundii, "dwardsiella tarda, aeromonas hydrophilla, (almonella, sp., (chewanella putrifaciens, and (treptococcus sp. It was also found out that Taro, !alamansi, 8otu Hola, and %orinda were the most active antimicrobial plants M?ee, et.al, =::LN. Another study conducted by him on the antimicrobial property of =-hydro/ypropane-D,=,F-tricarbo/ylic acid isolated from !alamansi "/tract. It was found out in his study that the crude e/tract and its bioactive component might have potential as antimicrobial agent for aquaculture use M?ee, =::7N. )'etrieved from httpC22www.soyuzfoods.com.ph2benefits.htm*

Chapter 9 RESEARCH METHODOLOG: Re&earch De& 'n +escriptive method was used and the e/perimental design specifically >arallel-group design was utilized throughout the study to serve as control for comparative study on the different hand sanitizers and the !alamansi fruit e/tracts and Talisay leaves e/tract.

Loca!e o, the Stud.

The e/periment was conducted at the +epartment of (cience and Technology ?aboratory Testing !enter. The study covered the months of Eune-$ctober =:DF.

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Te&t Mater a!& The materials used in the e/periment include three different products of commercialized hand sanitizers namely D* (afeguard =* 8reen !ross and F* #ygieni/, and plant e/tracts namely D* Halamansi )Citrus microcarpa bunge* fruit e/tract =* Talisay )Terminalia catappa* leaves e/tract and with B:; ethyl alcohol that served as a control, broth cultures of bacteria e.g. Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, an incubator set at FB degree !elsius, plane agar, sterile >etri dish, filter paper disc, sterile cotton swabs and forceps, bacticinerator.

Data Gather n' Procedure Pha&e 11Preparat on 6Ethano! c e(tract on o, dr ed Ta! &a. !ea-e& and e(tract on o, Ca!a#an& !ea-e&7 The fresh !alamansi fruits were washed with tap water and were e/tracted by squeezing to get the .uice and stored in air-tight container. @or ethanol e/traction, 9:g of air dried Talisay leaves powder was e/tracted with ethanol )J:-9:o!* in an e/tractor for DL-=J hours and solution was evaporated to dryness under reduced pressure and controlled temperature by using roto evaporator. The e/tract was stored in a refrigerator at J O! in air-tight bottle. 9:g2FIml concentration was used. Pha&e II1 E-a!uat on& o, Ant # cro+ a! act - t. o, natura! !ea, e(tract 6Paper D &% D ,,u& on Method7

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Antimicrobial activity of each plant e/tract was determined using a wattman paper dis diffusion method. The watmann paper dis was soa ed in the !alamansi and Talisay e/tract and allowed to stand for I minutes. >repare the assay plates with nutrient agar to be tested. 4sing a sterile loop, emulsify a colony from the plate. +ip the swab into the broth culture of the organism. 8ently squeeze the swab against the inside of the tube to remove e/cess fluid. 4se the swab to strea in a nutrient agar plate for a lawn of growth. This is best accomplished by strea ing the plate in one direction, then strea ing at right angles to the first strea ing, and finally strea ing diagonally. "nd by using the swab to strea the outside diameter of the agar. 0ith the used of forceps, pic ed out one paper disc and lay the moistened filter disc gently on the agar medium< Arranged the succeeding discs in much the same way as the agar wells< Incubate the plates inverted for DL to =J hours.

E-a!uat on o, ant # cro+ a! act - t. o, the co##erc a! 0ed hand &an t 0er&

The researchers inoculated the plates by dipping a sterile swab into the standardized inoculums2subculture. 'emove e/cess inoculums by pressing and rotating the swab firmly against the side of the tube above the level of the liquid. #old the >etri dish with the left hand, with the cover side up. 0ith the thumb and little finger, raise the lid and proceed with the inoculation. (trea the swab all over the surface of the medium tracing a zigzag course from side to side until reaching the middle and widest part of the plate. 'otate the plate through an angle of DL:O and continue to inoculate the second half of the pate in the same manner. After the inoculums have dried for F-I minutes, the researchers placed the paper discs dipped with the different hand sanitizers and the control by the use of sterile forceps.

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After the preparation, the plates were placed in an incubator set at F9O! for =J hours. After incubation, the plates were carefully inspected. 'esults were analyzed by measuring the zone diameter using a millimeter scale. %easurement was done on the undersurface of the plate without opening the lid.

Ta+!e D. Standard D a#eter o, /one o, Inh + t on 6##7 Very Active Active >artially Active Inactive PD7mm DJ-D7mm D:-DFmm QD:mm

The table showed the standard zone of inhibition which are categorized as Very active, active. >artially active and inactive .Very active category with a zone of inhibition of PD7 mm means that under this category, Test organisms are susceptible to the test samples, and so with the active. In the inactive with a zone of inhibition of QD: mm this means that test organism are sensitive to the test samples. Stat &t ca! Treat#ent

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All data gathered from the sensitivity testing for the control and e/perimental group was sub.ected to statistical evaluation by 4nivariate Analysis of Variance for interpretation with pQ:.:D level of significance.

Chapter ; PRESENTATION8 ANAL:SIS AND INTERPRETATION O" DATA This chapter represents the analysis and interpretation of data obtained after accomplishing the research methodology which deals with evaluating the antibacterial effectiveness of the hand sanitizers namely (afeguard, 8reen cross and #ygieni/ and the natural plant e/tracts which is the !alamansi )Citrus microcarpa bunge* fruit e/tract and the Talisay )Terminalia catappa* leaves e/tract against the test organism such as Staphylococcus aureus, Escherichia coli and Streptococcus and compared to the control which is B:; ethyl alcohol. The data below represent the results of the e/perimentation.

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Table =. Ana!.& & o, *ar ance n the d ,,erence n the 0one o, nh + t on o, the d ,,erent t.pe& o, +acter a and d & n,ectant& u&ed1

(ource Types of bacteria +isinfectant 4sed Typesof bacteriaR +isinfectant used "rror Total !orrected Total

df = I D:

%ean square D=.79F BL.FJD F==.:BJ

@ =L.::: D97.=D9 97I.9L:

(ig. .::: .::: .:::

F9 IJ IF

.J9F

Table = showed that the types of bacteria have a significant difference with a significance value of :.:::. And so with the disinfectant used with significance level of :.:::. These tables also showed that there is an interaction effect between the types of bacteria and the disinfectant used as shown by the value of the significance of :.::: at :.::D level of significance.

Table F. Ana!.& & o, *ar ance n the d ,,erence n the 0one o, nh + t on o, the d ,,erent t.pe& o, +acter a1

Types of bacteria ". coli >. aeruginosa

%ean DB.BBL DL.FFF

(td. "rror .D9: .D9:

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(. aureus

D9.99B

.D9:

Table F shows the zone of inhibition between the different types of bacteria. . P. aeruginosa has the largest zone of inhibition with a mean of DL.FFF. E. coli has a mean of DB.BBL which is smaller than that of P. aeruginosa. The smallest zone of inhibition is the S. aureus having only a mean of D9.99B. In these table >seudomonas aeruginosa is the disinfectant used as shown by its zone of inhibition. most sensitive bacteria to all the

Table J. Ana!.& & o, *ar ance n the d ,,erence n the 0one o, nh + t on o, the d ,,erent d & n,ectant& u&ed1 +isinfectant used Alcohol control !alamansi 8reen cross #ygieni/ (afeguard Talisay %ean =D.DDD DI.JJJ =D.DDD DB.LL7 DI.II9 DJ.JJJ (td. "rror .==B .==B .==B .==B .==B .==B

Table J shows that 8reen !ross hand sanitizer is the disinfectant used that is active against the three tested organism as shown by its zone of inhibition with a mean of =D.DDD. 8reen cross hand sanitizers e/hibit the largest zone of inhibition which implied that it is the most effective among the disinfectant used.

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Chapter < SUMMAR:8 CONCLUSION AND RECOMMENDETION The ob.ective of the study was to compare the effectiveness of the different hand sanitizers and the two natural plant e/tracts, the !alamansi)Citrus microcarpa bunge* fruit e/tract and Talisay )Terminalia catappa) leaves e/tract against "scherichia coli, >seudomonas aeruginosa and staphylococcus aureus using antimicrobial susceptibility testing. The +ried Talisay leaves were processed into "thanolic e/tract , !alamansi e/tract was e/tracted by simply squeezing the fruits to collect the .uices which was used and the three commercialized hand sanitizers namely, 8reen cross, (afeguard and the #ygieni/. The e/tents of the effectiveness of this were tested using >aper disc diffusion test with B: ; ethyl alcohol as the positive control. The result of the 4nivariate of variance showed that there is significance in the zone of inhibition in the different types of bacteria. And there is also an interaction effect between the type of bacteria and the disinfectant used.

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Conc!u& on

Aased on the findings of the study, the following were drawnC

The green cross hand sanitizer showed the largest zone of inhibition of the entire three test organism with a mean of =D.DDDmm. $n the other hand >seudomonas aeruginosa is the most sensitive test organism in the disinfectant used with a mean of DL.FFF.

!omparing the different hand sanitizer to the two plant e/tract tested against the three different test organism, the hand sanitizers showed the most effective and most active inhibitory effect.

Aased on table =, there is significant difference in the zone of inhibition of the different types of bacteria.

There is a significant difference in the zone of inhibition of the disinfectant used There is an interaction effect in the zone of inhibition between the types of bacteria and the disinfectants used. Reco##endat on&

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Aased on the previous findings gathered from the e/perimentation and conclusion of the study, the researchers suggest the followingC The future researchers to the study the antibacterial property of the hand sanitizes, !alamansi )Citrus microcarpa bunge) and Talisay )Terminalia catappa* e/tracts against other microorganism. Kuantitative determination of the different hand sanitizers and so with the !alamansi and Talisay e/tracts. Apart from the antibacterial property, antiparasitic activity of the !alamansi fruit and Talisay leaves e/tract should also be e/plored. @urther quality should carry out on the entire procedure.

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