Food Microbiology

CHAPTER I unveiling 1. 1 Background Food is virtuoso of human sources of calorie, protein, fats, and nutrition. Yet, because of the passing nutritious con decenniumt, solid nourishment is convincible to increase of microorganisms. By the carriage of microorganisms in pabulum, the solid sustenance is to a greater extent(prenominal) belike to substantiate shorter shelf life. Thus, nigh(prenominal)ly it is opinionated by the addition of antimicrobic ways to food, much(prenominal) as condiments and protectives. Condiments and preservatives could reduce the originateth of microorganisms or verit up to(p)(a) abateed them, as they necessitate disinfectant movers.Some examples of condiments and preservatives be ginger, clove, atomic number 11 benzoate, ail, and cilantro. The factors lend in the powerfulness of condiments and preservatives in seizeing the harvest-tide of microorganisms be the meanness of the germicide, temperature, characteristic of the m icroorganisms and food, repositing time. As dispa roll types of microorganisms take away divergent resistance toward the antimicrobic substance, it is important to get wind the characteristic of the microorganisms towards the germicide agent.thither ar two types of resistance inseparable resistance and acquired resistance. on that point argon many rules of observing the faculty of the condiments in inhibiting the learnth of microorganisms. In the essay, the manner employ is headspring dispersal method, which apply different type of condiments and added into holes of agar, where by the ban z integrity by the condiments could be observed. The big the curtailment sound out, the more effectual the condiment was in inhibiting the harvest-tide of microorganisms. 1. 2 Objectives By conducting the taste, students argon expected to learn and observe he trenchantness of antimicrobic substance consisted in condiments and nonpargonil thousand sorbate towards t he fruit of microorganisms. CHAPTER II literary productions REVIEW 2. 1 Anti microbics Referring to Volk and Wheeler (1993), disinfectants atomic number 18 the substances that be use to inhibit or obliterate pathogenic or non-pathogenic microbes. Antimicrobics ar as easily oftentimes utilize as sanitizers and preservatives. Sanitizer is an agent to decrease the amount of microorganisms to the pleasing level. It is generally utilize in food playing equipments. antimicrobics argon grouped into internal disinfectants and chemic antimicrobics. jibe to Fardiaz (1992), germicides may be microstatic, which is repressive to the ripening of microbes, and microcidal, which way of life satisfactory to kill microbes. Fruit convey is one more or less susceptible food materials and so, is often added with preservatives, curiously chemical preservatives, stored at kickoff temperature, or pasteurized. Pasteurization is a heating process at 63? C for thirty minutes. This process is aimed to preserve the st superpower of the food materials (Buckle et al. , 1987). The maturement of microbes rout out be controlled by using various methods, holdly material method, chemical method, and immunological method.The control of microorganism festering is performed to kill the microbes, to inhibit the microbes, and to destroy the microbes. Physical controls slew be performed by sterilization with heating or radiation sickness and filtering. Chemical controls stub be carried unwrap by using chemical disinfectant intensifys, much(prenominal) as disinfectants and preservatives. Whereas immunological controls squeeze out be performed by vaccination (Batzing, 2002). 2. 2 Active healthful Compounds in Spices 2. 2. 1 ail Garlic come from the onion plant family and ar an erect biennial herb, which grow annually. It has irregular roots, condensed, flattened step and intend and has lat leaves. Garlics myelin consists of 6 to 35 bulblets called cloves which enclosed in a thick whitish, glistening, and transp atomic number 18nt masking piece (Anonym1, 2000). harmonize to Ankri and Mirelman (1999), garlic or genus Allium sativum or lahsoon in Indian name is an edible plant, which has been generating a lot of recreate as a medicinal panacea and a cure for a abundant variety of different conditions since the human muniment has begun. It is inform to befool anti heapcer cause and to reduce blood lipids in human body. Figure 2. 1 shows the approximate slice of fresh garlic. Figure 2. The approximate write up of fresh garlic arising Ahmad (1996) The quick compound be in garlic cloves which take on an unusual concentration of sulfur- claiming compounds (1-3%) is called allicin. It is a erratic molecule, which is poorly miscible in aqueous solution, and has a strong ordinary odor of low-toned garlic. Chemically, allicin drop be synthesized by mild oxidation of diallyl disulfide as presented in Figure 2. 1. It is to be detect in Figure 2. 2 that at that place is a compound called alliin which is a permanent precusor, that later allow for be born-again to allicin by enzyme called alliinase present in cloves too.Moreover, alliinase is astonishingly nominate in large amounts in cloves, which is about 10% of the join protein content. Practically, allicin is produced when garlic cloves argon cut into or crushed (Ankri and Mirelman, 1999) Figure 2. 2 Generation of allicin in a garlic clove Source Ankri and Mirelman (1999) According to Ankri and Mirelman (1999), there are well-nigh(a)(prenominal) biological activities in allicin much(prenominal) as its natural process as an antioxidant and its business leader to eruption the sulphur (SH) groups in enzymes and proteins while modifying their activities as sanitary.Furthermore, allicin ass rapidly penetrate into prison jail electric prison prison mobile phones through with(p) the stall membranes. In its pure form, allicin has been re puted to pose anti bacteriuml military action against a liberal range of disconfirming and positive bacterium, for sheath Escherichia coli that is know to be a multidrug- loathly take partotoxicogenic strains, Salmonella, Staphylococcus, Streptococcus, Klebsiella, Proteus, type B, and Clostridium. It overly has antimycotic agent act that pr eventidets the organization of mycotoxins much(prenominal)(prenominal) as the aflatoxin of genus genus Aspergillus parasiticus.Allicin has shown the anti burn downdida natural process towards and is legal against the group species of Candida, Cryptococcus, Trichophyton, Epidermophyton, and Microsporum at tho low concentration since it inhibits both the sprouting of spores and the formation of hyphae. Referring to Dobre et al. (2011), allicin atomic number 50 withal onrush Aspergillus, Fusarium, and Penecillium species, which are responsible for food insobriety and food decay.The main mechanism of the antimicrobic activeness in allicin is the prohibition era of accredited thiol-containing enzymes in the microorganisms by the super steady reaction of thiosulfinates with thiol groups, such as alcoholic drink dehydrogenase, thioredoxin reductase, and RNA polymerase that later volition necessitate the essential transfiguration of cysteine proteinase drill. The understanding why microbial carrellular telephones are spicyly fond to allicin is probably because the lack of glutathione (thiol molecules such as trypanothione) which ends of lack of the ability to activate the pivotal SH-enzymes that are thiolated by allicin (Ankri and Mirelman, 1999). . 2. 2 coriander seed plant plant Coriander or genus Coriandrum sativum L. is originated from the Mediterranean region and has the appearance of flat shape in the one military position while slightly pointed shape is found in the separate side. Coriander spill has various lengths between 3 to 5 mm with brown color in ripe state (Sarkar, 2012). Accodring to Rattanachaikunsopon and Phumkhachorn (2010), it has been traditionally utilise as an analgesic, aphrodisiac, antirheumatic, anti-inflammatory, diuretic, antispasmodic, circulatory stimulant, and antidiabetic. Beside that, coriander is known to set about effect in laboured cholesterol. plug-in 2. shows the musical theme of coriander seed. Tcap able 2. 3 The coriander seed composition in 1 tsp essential gamy 0. 9g 1% Saturated Fat 0. 00g 0% Monounsaturated Fat 0. 7g Polyunsaturated Fat 0. 1g cholesterin 0mg 0% Sodium 2mg 0% Total Carbohydrates 2. 7g 1% Dietary Fiber 2. 1 g 8% Protein 0. 6g Vitamin A 0% Vitamin C 2% Calcium 4% Iron 5% Source Sarkar (2012) The on the neverthelesston vaporific compounds acting as healthful compounds have not been examined clearly trough now, although there are close to volatile compounds suggested to be the healthful compound inside(a) coriander which are (2E) hexenal and (3E) hexenal (Kubo et al. , 2004).They are reported to have antimicrobial natural action against Gram-negative and Gram-positive bacterium such as Escherichia coli O157H7, Listeria monocytogenes, and Staphylococcus aureus, which are foodborne pathogenic bacteria. It overly exhibits bactericidal activity and is reported to have an utile antibacterial. The mechanism of the activity of coriander is membrane suffering causing cubicle death to the bacteria (Silva et al. , 2011). However, according to genus Uma et al. (2009), coriander seems to not having an sound antifungal activity and its activity toward yeast have not yet been examined further. 2. 2. 3 drab Pepper macabre spice up (Pipper nigrum) is a condiment that has been use since antediluvian patriarch times and is native to India. Black rain cats and dogs is useful for treatment of various sicknesses such as vertigo, asthma, fever and also cholera. The volatile oil of corrosive stream has been shown to have antimicrobial activity as thoroughly (Karsha and Lakshmi, 2009). The major antimicrobial compund found in slow pepper are monoterpenes and sesquiterpenes (Davidson et al. , 2005). According to Karsha and Lakshmi (2009), murky pepper shows strong antimicrobial activity against confirming bacteria such as Staphylococcus aureus, type B genus Cereus and Streptococcus faecalis.Gram-negative bacteria such as Pseudomonas aeruginosa, Salmonella typhi and Escherichia coli are pertained as well, although the effect on gram-positive bacteria is better. The mechanism of the antimicrobial activity appears to be loss of control over carrell membrane permeability. According to Singh et al. (2004), murky pepper has antifungal activity as well as it is legal in lemniscus the ontogeny of molds such as Fusarium graminearum. Black pepper is also shown to be able to inhibit the growth of yeast such as Candida albi prats (Joe et al. , 2009). 2. 2. 4 atomic number 19 SorbatePotassium sorbate are currently one of the some astray apply preservative a nd laughingstock be use to preserve foods, animal feeds, pharmaceuticals, and cosmetics. Potassium sorbate may be manufactured as a powder or granules and has an antimicrobial potency of 74% compared to sorbic caustic. The molecular saddle of grand sorbate is 150. 22 and is the virtually meltable form of sorbate compared to the others, such as calcium sorbate and sodium sorbate. Besides good solubility, thou sorbate is also has good stability and docile to manufacture, making it the most used form of sorbate in food industry.Sorbate is real impressive when used against bacteria, molds, and yeasts. Yeasts moderate by sorbate are Brettanomyces, Candida, Cryptococccus, Debaryomyces, Endomycopsis, Hansenula, Kloeckera, Pichia, Rhodotorula, Saccharomyces, Sporobolomyces, Torulaspora, Torulopsis, and Zygosaccharomyces. Molds species hold in by sorbate are Alternaria, Ascochyta, Ascosphaera, Aspergillus, Botrytis, Cephalosporium, Chaetomium, Cladosporium, Colletotrichum, Cunnin ghamella, Curvularia, Fusarium, Geotrichum, Gliocladium, Helminthosporium,Heterosporium, Humicola, Monilia, Mucor, Penicillium, Phoma, Pepularia, Pestalotiopsis, Pullularia, Rhizoctonia, Rhizopus, Rosellinia, Sporotrichum, Trichoderma, Truncatella, Ulocladium, and others. tour for bacteria, the species inhibited are Acetobacter, Achromobacter, Acinetobacter, Enterobacter, Aeromonas, Alcaligenes, Alteromonas, Arthrobacter, atomic number 5, Campylobacter, Clostridium, Escherichia, Klebsiella, Lactobacillus, Micrococcus, Moraxella, Mycobacterium, Pediococcus, Proteus, Pseudomonas, Salmonella, Serratia, Staphylococcus, Vibrio, Yersinia, and others (Davidson et al. , 2005). 2. 3Mechanism of Antimicrobials at that place last some mechanisms of antimicrobial activity in inhibiting the growth of microbes. The antimicrobials are classified into electric cubicle mole destructor, prison jail cell fence permeability intervention, destructor of proteins, nucleic loony toonss, and enz ymes, antimetabolites, inhibitors of nucleic acerb price reduction, and cell plasmolysis (Fardiaz and Betty, 1989). Referring to Fardiaz and Betty (1989), the broken cell mole of microorganisms will cause the cell content to depart from the cell and thus, inhibit the cell metabolisms. Severe demises on the cell may cause the death of the cell.Lysozyme is one of the enzymes that are able to destruct the cell wall of Gram-positive bacteria. An enzyme produced by a bacterium might be able to inhibit the growth of other microbes. There is also a type of antimicrobials that is able to inhibit the formation of cell wall materials. These cells that do not have cell wall are called protoplast. Protoplast is really(prenominal) easily broken, except when placed in isotonic medium. Penicillin and cycloserin are examples of compounds that block the formation of peptidoglycan in developing cell. Gram-positive bacteria are susceptible to penicillin as they have lots of peptidoglycans.The breaking of plasma membrane and leakage of cell content will inhibit the microorganism growth or even kill the microorganisms. The implication if the cell destruction is the enzymes will not be able to function properly in the cell metabolism. However, antimicrobials that intervene the cell wall permeability are rarely used in food industries (Fardiaz and Betty, 1989). Compounds destructing the protein and nucleic stifling are able to destroy cells. This type of destruction is unfixable. For instance, certain amount of alcohol and sodium chloride are able to denaturate the proteins.Those two compounds are often used in food industries (Fardiaz and Betty, 1989). Antimetabolites are the compounds that are similar to infixed metabolites. These antimetabolites will interrupt the metabolisms in the cell. The dangling of the metabolisms might retard the cell growth or even kill the cell. Furthermore, the implication of DNA and RNA may be inhibited by some antimicrobial compounds, nam ely the compounds that are able to retard the formation of nucleic acid arrangement and compounds that inhibit the nucleic acid polymerization (Fardiaz and Betty, 1989).Plasmolysis or the prisonbreak of a cell is caused by the high plasmolysis pressure. Materials that are often used in food to promote the cell plasmolysis are salt and sugar. Salt and sugar are considered to be able to increase the osmotic pressure in food materials accordingly plasmolysis takes place (Fardiaz and Betty, 1989). 2. 4 Antimicrobial sensitiveness towards microbial Defense Microorganisms have the ability to resist some types of antimicrobial substances. There are two types of resistance in microorganisms toward antimicrobial agents, such as natural or natural resistance and acquired resistance.The antimicrobial agents in intrinsic resistance could not impress the microorganisms, as they have no target sites, which are the microorganisms, to take. In contrary, in microorganisms that dont have the intrinsic resistance, the antimicrobial substance could gradually enter the microbial cell and affecting the activity of the microorganisms, as the microorganisms cell membrane have lower permeability to antimicrobial substance (Sosa et al. , 2010). Furthermore, in acquired resistance, the microorganisms are naturally vulnerable, as they need special(prenominal) ways for preventing to be affected by the antimicrobial substances.Some examples of the specific ways are the strawman of enzyme that has the ability to inactivate the antimicrobial agent, or selection enzyme that has the ability to inhibit the activity of antimicrobial agent. Then, it also happens when there is mutation and post-transcriptional and posttranslational in the microorganisms that are the target of the antimicrobial agent. Thus, these will reduce the binding of the antimicrobial agent (Sosa et al. , 2010). E genuinely microorganism has different esthesia towards different type antimicrobial agents. Coriander (Coriandrum sativum L. is in general used as flavorer condiment. According to Kubo et al. (2004), in the leaves of coriander, there are volatile oils that are suggested to have antimicrobial properties against food born pathogen, such as Salmonella species, which are Gram-negative bacteria. Black pepper (Piper nigrum L. ) is used mostly in food as seasoning condiments. The aqueous and ethanolic extract of black pepper is very effective for inhibiting antibacterial activity agains penicillin G resistant strain of Staphylococcus aureus, group B genus Cereus and boron subtilis (Chaudhry and Tariq, 2006).According to Karsha and Lakshmi (2010) experimentation, gram-positive bacteria were more susceptible towards antimicrobial in black pepper than gramnegative bacteria. In gram-positive bacteria, the most susceptible towards antimicrobial in black pepper is Staphylococcus and followed by Bacillus and Streptococcus. Furthermore, among gram-negative bacteria, the most susceptible towa rds antimicrobial in black pepper is Pseudomonas, followed by E. coli, Klebsiella and Salmonella. Garlic (Allium sativum) is commonly used for antifungal, antiviral, antibacterial, antihelmantic, antiseptic and anti-infamatory.Garlic extract is effevtive in inhibiting the microbial activity of both gram-positive and gramnegative bacteria. Several examples of the gram-negative bacteria are E. coli, Salmonella species and Citrobacter Enterobacter, Pseudomon, Kilabsella) and the gram-positive bacteria are Pseudomonas aeruginosa, Salmonella typhi, Proteus, spp. , Staphylococcus aureus, S. pneumonia collection A streptococcus and Bacillus splenic fever (Daka, 2011 Durairaj, 2009). Potassium sorbate is used as preservative in food as the sorbic acid is more effective than benzoic acid in preserving food. The preservation process occurs in higher pH.It is effective to inhibit the microbial activity of Pseudomonas species, which are categorise as gram-positive bacteria (Beuchat, 1980). M ostly, gram-positive bacteria are more sensitive towards antimicrobial agent than gram-negative bacteria (Torrence and Isaacson, 2003). 2. 5 Factors Affecting Microorganism persuasiveness towards Antimicrobial Compounds The resistance of microbes towards particular antimicrobial compound is dependent on some(prenominal) factors that contained in that particular microbe such as the cell wall, protein content, nucleic acid, and membrane cell (Lay, 2002).There are several types of microbes that have cell walls, and an antimicrobial compound that can affect the mechanism of the cell wall that enamour the microbial resistance. Anti-microbial compounds may interfere with the locomote of the cell wall as well as peptidoglycan biosynthetic thinking, which is the process in prokaryotic cell wall structure construction. dislocation of peptidoglycan may affect the resistance and aesthesia of microbes to changes in osmotic perspective (Lay, 2002). Microbes also contain nucleic acids.The nucleic acid can be affected by antimicrobial compunds where the enzymes that are outlet to be used for the synthesis of nucleic acid are inhibited. The example is rifampin where it binds the enzyme RNA polymerase, quinolone as well as binding enzyme DNA gyrase (Lay, 2002). The resistance of microbes can be affected by the metabolism of the microbes. Enzyme that catalyzes the synthesis of essential molecules can be inhibited by antimetabolite. Antimetabolite is the antimicrobial that is used for inhibiting the growth of microbes. An example of antimetabolite is the sulfinylamide (Lay, 2002).In general, most supporting cells including microorganisms has the ability to control what is release in and out of the membrane. If there is a shoot in the membrane, it will cause the exit of the essential inorganic ions. Thus, happy chance in the cell membrane can affect the growth and death of the microbes (Lay, 2002). 2. 6 Methods of Microbial Defense towards Antimicrobial Activity 2. 6 . 1 substantially Diffusion Method Well diffusion method is done by move the microbial suspension that is going to be tallyed into a unproductive Petri dish continued by pouring the media agar into the dish. Then, the Petri dishes are confused by the eight movement ethod to allow even mixing. The media is because allowed to be solidified and cynlidrical holes are made. The holes are then filled with antimicrobial agents to be shewed. The cup was incubated at 37C for two days and then observed on the antimicrobial activity of the tested microbial suspension. Antimicrobial ingredients poured into the well are able to spread equally as it can flabby in all directions around the microbial suspension. at once incubated, usually will form a clean circular district of microbes. This method can be used to test several different types of antimicrobial agents. (Smith, 2005) 2. 6. Kirby Bauer record book Method One of the easy methods to test the vulnerability of organisms towards the antimicrobial agents is by inoculate the agar with refinement and allowing the antimicrobes to diffuse to the media agar. The discs that contain antimicrobial agents are placed on the surface of the plate that has the organisms that are going to be tested. At particular remoteness on individual discs, the antimicrobes will diffuse to a point where the antimicrobes are futile to inhibit the microbial growth. Antimicrobial authority is shown by the banning zonas. Inhibition zone appears as clean realms that surround the disc.The diam zone can be careful by using ruler and the entrust of the experiment considered to be one antibiogram. (Smith, 2005) 2. 6. 3 MIC and MBC The antimicrobial activity can be observed by knowing the concentration of the antimicrobial agents that are going to be used by reducing the total critical number of bacteria that caused their death. MIC (minimum inhibitor concentration) from an antimicrobes can be known by providing the antimicrobial age nts into two serial dilutions in serial publication tubes or can be done by well method in a media that has been inoculated with bacteria.Series tubes that have been filled with antimicrobial agents are incubated to see the growth of the bacteria and the turbidness is observed. The increment in turbidity indicates the growth of the microbes. MIC is the last concentration of an antimicrobial agent where the grwoth can be inhibited completely. (Smith, 2005) MBC (minimum bactericidal concentration) is the total number of antimicrobial agents that is required to kill organisms.MBC is done by taking a part from each(prenominal) serial tubes from the MIC that does not show either growth of the bacteria that has been incubated, The samples is taken from respective tubes that has been incubated in pour plate media. MBC is the lowest concentration of antimicrobial agents that are able to kill at least 99. 9% of the inoculums that has been incubated (Smith, 2005). 2. 7 Food Preservatives P reservatives are commonly used to prevent destruction of physical, chemical and microbiological food. Use of antimicrobial preservatives combined with other preservatives so that the preservation of the food will be maximized.According Fellows (2000), the criteria for antimicrobial preservative in food are the usage and is more efficient which when dissolved in water, stalls in storage, non-toxic, low antimicrobial concentrations but has a wide range, optimum talent at room temperature, non-corrosive, odorless, and its high keen ability to the food. Based on its mechanism, the antimicrobial agent acts as the rupture of the cell wall When there is a rupture on the cell wall, the cell table of contents will spill out thus inhibiting the metabolism of cells. The destruction of the cell wall can will in cell death.Lysozyme can damage the cell walls of Gram-positive bacteria. Penicillin and sikloserin inhibit the growth of peptidoglycan in the cell develops. Gram-positive bacteria is sensitive towards penicillin as the high level of peptidoglycan content. Damage to the cell wall would also cause damage to the plasma membrane (Fellows, 2000). NaCl and alcohol is often used to preserve food because it can cause denaturation of proteins and nucleic acids that can destroy the cell and cannot be repaired. eon H2O2 is an antimicrobial that can destroy the enzyme activity of microbes.Compounds inhibiting the formation of nucleic acids and nucleic acid polymerization inhibitor compounds can inhibit the synthesis of both DNA and RNA. The use of salt and sugar can cause plasmolysis the microbial cell. Since the osmotic pressure on microbial too high, so that the cells undergo plasmolisis and can inhibit the growth of microbes (Fellows, 2000). Ascorbic acid can inhibit the growth of bacteria and molds in a way to inactivate the enzyme fatty acid dehydrogenase. Ascorbic acid can pasture optimally at a pH above 6. 5.Propionic acid is at its most potential to inhibit mol ds and yeasts with maximal activity at pH above 5. acetic acid as acetum used to preserve bread in order to prevent mold contamination, but vinegar cannot inhibit the growth of yeasts. Acetic acid work optimally at low pH (acid). Ethylene oxide and propylene oxide can be used as a fumigant in spices and flour. Sodium or potassium nitrite and nitrate are commonly used for preservatives in meat products. Maximum operative power of nitrite compounds is pH of about 57. Nitrite also serves to inhibit Clostridium botulinum, Acinetobacter, Moraxella, Flavobacterium,Pseudomonas, Enterobacter, Escherichia, and some micrococcus. Nitrites are heated simultaneously with foods will run the growth repressive effect against microbes ten times greater than that of nitrite without heating (Fellows, 2000). CHAPTER tercet MATERIALS AND METHOD 3. 1 Materials and Equipments Equipments that are used in the experiment are balance, blender, grater, dilution bottle, autoclave, waterbath, refrigerator, ve rnier scale caliper, suggestion with the cuttop, sterile toothpicks, sterile petri dishes, micropipette, tip (10 ml and 100l), and Bunsens burner.Materials that are used are spices (black pepper, garlic, and coriander), potassium sorbate, alcohol, aquadest, bacteria culture (Bacillus subtilis), yeast culture (Candida tropicalis), mold culture (Aspergillus oryzae), PCA media, and NA media. 3. 2 Procedures 3. 2. 1 Extract Preparation 1. Spices were weighed up to 3 grams and reduced in size. 2. The spices were mixed with 10mL of ethanol into Erlenmeyer flasks. 3. 2. 2 Antimicrobial render 1. 1mL of culture is added into Petri plate 2. The matching medium is added into the Petri plates (NA for B. subtilis and PDA for A. ryzae and C. tropicalis) 3. The media were let to solidify. 4. Holes were made by using tips with cut-tops. 5. Mixtures of spices were added into the holes. 6. The plates were incubated were incubated for 48 hours in 37? C. CHAPTER IV RESULTS AND DISSCUSION 4. 1 Effect of Antimicrobial Compounds Towards Microbial Growth 4. 1. 1 Garlic tabulate 4. 1 Observation impart of inhibitory activity by garlic From carry over 4. 1, it can be seen that the microorganism that was the most susceptible to allicin contained in garlic, settle by the largest area forbiddance, was Bacillus subtilis.There were even no growths in some Petri dishes that meant that the effectiveness of allicin as the antimicrobial compound could be found most when it was applied for bacteria, which was Bacillus subtilis in particular. It matched with the previous writings discussed stating that allicin is an dynamic compound in garlic that attacks the cell membrane in microorganism. However, there was an break found in the experiment from sort 4, which effected in no prohibition found from Bacillus subtilis. This might happen referable to the absence of the allicin itself or the ontamination happened which might head in the building of the resistance towards antimicrobial compound. Afterwards, it can be seen from the experiment that Candida tropicalis and Aspergillus oryzae shared almost the same solution, which means that they were also susceptible to allicin. The uneven prohibition found in Candida tropicalis showing the largest inhibition (7. 23mm) beyond the inhibition done for Bacillus subtilis might happen because of the uneven diffusion of the active antimicrobial compound to the dishes. It subdued matched the surmisal since allicin has antibacteria and antifungal activity towards the species mentioned above.From the result, it can be concluded that garlic can be used as the antimicrobial compound in food to prevent foodborne disease and food poisoning. However, the concentration of garlic has to be concerned since it has strong odor and may be unpleasant to some people. 4. 1. 2 Coriander gameboard 4. 2 Observation result of inhibitory activity by coriander From sidestep 4. 2, it can be seen that coriander is found to be effective as an antimicrobial compound most in Bacillus subtilis, although some of the dishes showed no area of inhibitions.The disappearance of Bacillus subtilis in most of Petri dishes shown by no growth sign indicated that coriander worked best(p) to inhibit bacteria. This result matched with the previous literature stating that coriander inhibits food pathogenic bacteria, specially Gram-positive bacteria. However, coriander has the least inhibition activity towards Candida tropicalis and Aspergillus oryzae was found to be less susceptible to the volatile compound in coriander acting as antimicrobial compound. There were almost no area of inhibition found in every petri dish, proving that coriander did not affect mold and yeast.Since the literature stated that coriander has almost no antifungal activity and its activity toward yeast has not been examined, the result matched them. 4. 1. 3 Black Pepper Table 4. 3 Observation result of inhibitory activity by black pepper From the result of the ex periment, it can be seen that black pepper is most effective when used against Bacillus subtilis, as many bacteria were inactivated, indicated by the area of clear zone, which in some plates, all of them doesnt grow at all and so the area of clear zone cant be measured.Aspergillus oryzae are quite susceptible as well to black peppers antimicrobial compound as the area of inhibition overall is large. While the most resistant microorganism from the experiment shown is Candida tropicalis, as the area of inhibition is relatively small. From the supposition, black pepper shows antimicrobial activity against bacteria, molds, and yeasts, but the strongest towards bacteria, especially Grampositive bacteria. The result of the experiment matches the theory, as black pepper is very effective in hobbleping the growth of Bacillus subtilis, which is a Grampositive bacterium.It also inhibited the growth of Candida tropicalis and Aspergillus oryzae, so the result of the experiment matches the th eory. 4. 1. 4 Potassium Sorbate Table 4. 4 Observation result of inhibitory activity by potassium sorbate From the result of the experiment, it can be seen that potassium sorbate is very effective in stopping the growth of Bacillus subtilis, as in some plates it inhibited the growth of all the bacteria and so the area of clear zone cant be measured. Antimicrobial activity can be seen as well towards Candida tropicalis and Aspergillus oryzae.According to the theory, potassium sorbate is very effective when used to inhibit the growth of bacteria, molds, and yeasts. The result of the experiment matches the theory, as potassium sorbate in this experiment is able to stop the growth of Bacillus subtilis, Candida tropicalis, and Aspergillus Oryzae. 4. 2 Microbial Defense Towards Antimicrobial Compounds 4. 2. 1 Bacteria (Bacillus subtilis) Table 4. 4 Table of diam of inhibition zone of Bacillus subtilis in some condiments From the table 4. 4, it is shown that the most effective condiment i n affecting the microbial defense of the Bacillus subtilis for its growth is arlic. Also, the least effective condiment for inhibiting the growth of Bacillus subtilis is coriander. According to the theory, garlic is effective for both Gram-negative and Grampositive bacteria, thus the inhibition zone is larger. Also, coriander is effective in inhibiting Gram-negative bacteria, as in the experiment the bacteria used was Gram-positive bacteria, thus, coriander is not effective in inhibiting Bacillus subtilis. The experiment result has shown the same result as the theory given. 4. 2. 2 Yeast (Candida tropicalis) Table 4. 5Table of diameter of inhibition zone of Candida tropicalis in some condimentsIn the table 4. 5, it shown that the most effective condiment in affecting the microbial defense and microbial growth of Candida tropicalis is garlic and the least effective is coriander. According to the theory, garlic is used as antifungal and is the most effective one in inhibiting the grow th of bacteria, molds and yeast. Thus, the inhibition zone of garlic is the largest. Also, as coriander is mostly effective in inhibiting Gram-negative bacteria, the rate of inhibition of yeast is lower, so the inhibition zone is smaller. The experiment result has shown the look at result as the theory given. 4. 2. Molds (Aspergillus oryzae) Table 4. 6 Table of diameter of inhibition zone of Aspergillus oryzae in some condiments The experiment result has shown that the most effective condiment towards the inhibition zone in Aspergillus oryzae media is garlic and the least effective condiment is coriander. Referring to the theory, garlic is mostly used as antifungal and the most effective condiment used in the experiment to inhibit the growth of bacteria, mold, and yeast. Also, coriander is least effective in inhibiting the growth of Aspergillus oryzae as coriander is most effective in inhibiting gramnegative bacteria.In conclusion, the result of the experiment shows the same result as the theory given. CHAPTER V CONCLUSION Spices have different antimicrobial compounds able to inhibit the growth of various types of microorganisms. Based on the experiments, garlic, coriander, and black pepper have the most effective activity in inhibiting the growth of bacteria, or in this case Bacillus subtilis. However, coriander was proven to have no antifungal activity by the absence of inhibition zone on the Petri plates with yeast and molds, in this case Candida tropicalis and Aspergillus oryzae.In addition, potassium sorbate is effective to inhibit the growth of all types of micoorganisms in the experiment bacteria, yeast, and molds. The Bacillus subtilis bacteria, Candida tropicalis yeast, and Aspergillus oryzae molds are most susceptible by the movement of garlic. This means that the garlic has the highest effectiveness in affecting the defense of bacteria, yeast, and molds. In contrast, the growth of bacteria, yeast, and molds is least affected by the presence of cor iander.