IDENTIFICATION OF COLIFORM BACTERIA IN IN FOOD STALLS AROUND GORONTALO CITY SNACK MARKET

Ana Pepiana1), Laksmyn Kadir2), and Agusrianto Yusuf3) 1,3) Bina Mandiri University of Gorontalo 2) State University of Gorontalo Email: [email protected]

ABSTRACT Unsafe food will be the cause of health problems in the community. Chili sauce is a sauce based on chili that is crushed until the water content comes out so that it appears spicy. Chili sauce is processed in a simple way so that it can trigger contamination of microorganisms, one of which is coliform bacterial contamination. This research aims to look at the contamination of coliform bacteria in Chili sauce offered to eat around the snack market of Gorontalo City. This research is a descriptive study with a qualitative approach and examination of Coliform bacteria using the Most Probable Number (MPN) method. The population in this study was 14 samples with Accidental Sampling techniques. Coliform bacterial test results showed that in Chili sauce samples found positive results contaminated with Coliform bacteria, namely 11 samples with a percentage of 78.6% and negative results as many as 3 samples with a percentage of 21.4%. Based on the results can be concluded that a positive result was found 1 sample from the fecal coliform bacterial group in this case suspected Escherchia coli bacteria and 10 samples from the nonfectual coliform bacterial group. Keywords: sambal, coliform bacteria, food stalls

INTRODUCTION marketers in accordance with its existence Food Stalls is a place that sells food then food stalls should be able to provide and drinks. Food stalls are located in a services or provide food and drink needs public environment. A food stall is a to good food stall consumers will be able building that closes with equipment used to cause comfort to consumers so that it for the manufacturing and sale or can also be profitable for food stalls [5]. presentation of food and beverages to the The requirements are stated in the public, where the process of making and Decree of the Minister of Health of the selling or serving food is intended for the Republic of No. general public and the way it is presented 715/Menkes/SK/V/2003 on Jasaboga at a certain time [8]. Sanitary Hygiene Requirements covering The quality and safety of the food the location, buildings and facilities of provided in the food stall must be disposal and management of construction maintained because every food processing waste, floors, walls. lighting of treatment process contains potential contamination plants and clean water sources. Factors to that needs to be controlled to ensure the note in order to be able to organize safety of food consumed, therefore it is effective food sanitation are food factors, necessary personal hygiene in food human factors, and equipment factors.

11 Identification of Coliform Bacteria in Chili Sauce in Food Stalls around Gorontalo City Snack Market

Food is a source of human energy in health facilities was 4,274,790 sufferers order to be active. Food that is well and there was an increase in 2018 to consumed is a healthy and safe food. Food 4,504,524 sufferers or 62.93% of the is one of the three basic sources of need estimated diarrhea in health facilities. The for human life in addition to food and incidence of diarrhea of all ages nationally food. Sandang and food are a basic human is 270/1,000 inhabitants [9]. necessity because they are useful for Prevelence of diarrhea in Indonesia in providing protection for each other [6]. 2017 the number of people with diarrhea Bacterial contamination of food of all ages (SU) served in health facilities indicates the risk of various food-borne as many as 4,274,790 sufferers, and there diseases that are harmful to public health was an increase in 2018 to 4,504,524 and should be sought solutions. Many sufferers or 62.93% of the estimated studies state that Escherichia coli causes diarrhea in health facilities. The incidence diarrhea. There are now 4 groups of of diarrhea of all ages nationwide is Escherichia coli that can cause diarrhea, 270/1,000 inhabitants. The Central Bureau namely ETEC, EPEC, EIEC, and EHEC. of Statistics of Gorontalo Province (2018) Humans can be exposed to these bacteria also recorded 22,117 cases of diarrhea and when consuming food or drink that has in Gorontalo City there were 3,189 been contaminated by faeces from the cases. This diarrhea is caused by livestock. Escherichia coli is dangerous contamination of microbial agents, because it produces toxins commonly namely coliform bacteria in food known as shiga toxins that can penetrate consumed [7]. the intestines and interfere with the Coliform bacterial contamination due function of other organs. Food Safety to lack of hygiene and sanitation in food needs to be considered in order to improve such as Chili sauce processing. Chili the degree of health and avoid the degree sauce is a companion food that is part of of disease [12]. the eating culture known to Indonesians Foodborne disease isacommon public from Sabang to Merauke. Chili sauce can health problem. Around the world there be served in conjunction with other foods are millions of people due to foodborne as flavorings. Each region has different diseases. Each year, there are Chili sauce recipes that are often added approximately 1,500 million cases of with local plants known in the area. Chili diarrhea and an estimated 70% of cases of sauce is often added with famous diarrhea disease occur due to foodstuffs in certain areas. This is what contaminated food. One of the bacteria makes Chili sauce have a distinctive taste that can cause foodborne disease is in each region [14]. coliform bacteria that can have an impact The addition of chili sauce as a on diarrhea disease [12]. companion food is able to invite appetite Diarrhea disease is still a public health and reduce the taste of blandness in problem in developing countries such as food. Chili sauce is a product that has in Indonesia. The target coverage for been very well known by the public people with diarrhoea of all ages (SU) because it is able to accompany almost all who come to health care is 10% of the kinds of processed foods. The high level estimated number of people with Diarrhea of Chili sauce consumption makes SU (Incidence of Diarrhea SU multiplied consumers want to consume it in a by the number of residents in one working practical form. This makes Chili sauce area within one year). In 2017 the number often used as a business opportunity in the of people with SU diarrhea served in form of instant Chili sauce [11].

12 Journal of Health, Technology, and Science (JHTS) Ana Pepiana E-ISSN: 2746-167X, Vol. 1, No. 2, December 2020

Inappropriate processing practices can contamination in food with be seen from hiegene food cookers, APM Coliform maximum 100/g. sanitary equipment and foodstatives used, Based on that background, researchers are as well as the location of selling will also interested in conducting research with the title affect the quality of the chili sauce [15]. of identification of coliform bacteria in Chili Food commonly sold in food stalls sauce in restaurants around gorontalo city around Gorontalo city is usually served snack market. with additional chili sauce. Chili sauce is a sauce based on chili that is crushed until RESEARCH METHODS the water content comes out so that it The type of research used is appears spicy. There are various variations descriptive observative with a qualitative of Chili sauce. Each variety demands a approach, i.e. researchers observe directly variety of ingredients and condiments as the objects to be studied, then described well. Although the simple process of descriptively to know the or not of making chili sauce can’t be taken coliform bacteria in Chili sauce in food lightly. All seasonings, ingredients, and stalls around the city's snack market ways of making them should be properly Gorontalo. The number of samples as considered. That resulted in a delicious many as 14 Chili sauce samples taken spicy taste [13]. by accidental sampling method. The fact in the field is found that the Coliform bacteria identification data people of Gorontalo love food that has a is obtained through testing in the spicy taste, without thinking about the microbiology laboratory of Universitas quality of the Chili sauce itself. In the Bina Mandiri Gorontalo using mpn process of making Chili sauce is made method (Most Probable Number)series 3 simply. In addition to the simple way of tubes. Isolation stage and identification making it, the Chili sauce in this food stall stage using 3 stages of testing namely, has also been served on the dining table, identification test using Lactose so these people do not know if this Chili Broth media (LB), second stage booster sauce has good quality or not. test using Eosin Methylen Blue From unknown quality and simple Agar (EMBA) media, and third stage way of making it can certainly be one of complementary test by performing gram the factors of declining public health level coloring technique. On identification tests when consuming Chili sauce that is easily and booster tests for recent results use polluted by various types of micro positive and negative categories. organisms such as coliform bacteria [13]. The positive category if the results of Based on Indonesian National the test are proven to contain Standard (SNI) No. 12–7388–2009 Coliform bacteria by evidenced from the recommends the maximum limit identification test results using lactose of coliform bacterial contamination in broth media (LB) shows the appearance of Chili sauce with the most likely number noise and gas formation, and in the (APM) coliform which is < 3/g. Based on booster test showed bacterial growth in SNI 7388-2009 about the maximum the media Eosin Methylen Blue microbial limit on chili sauce for Agar (EMBA) with characteristics of MPN Coliform 100/ml. And Regulation of metallic green colonies for coliform the Head of the Food and Drug Control bacteria fecal group and colonies with Board of the Republic of Indonesia pink and transparent color with mucus Number HK.00.06.1.52.4011. About the for non-fecal coliform bacteria. For the maximum limit of microbial and chemical results of gram coloring examined in the microscope at 1000x magnification

13 Identification of Coliform Bacteria in Chili Sauce in Food Stalls around Gorontalo City Snack Market produces rod-shaped bacteria and is gram Sample 1  43 negative characterized by the result of red Sample 2  - gram coloring. This corresponds to the morphological characteristics Sample 3  150 of Escherchia coli bacteria and Sample 4  >2,400 Enterobacter aerogenes. The negative Sample 5  >2,400 category if the test results are not proven to contain Coliform bacteria in accordance Sample 6  >2,400 with the test results in the laboratory using Sample 7  - the Most Probable Number (MPN) Sample 8  4 method with Lactose Broth (LB) Sample 9  - and Eosin Methylen Blue Agar (EMBA) media. Sample 10  15 The data analysis was conducted by Sample 11  >2,400 comparing the results of coliform bacteria  identification testing on Chili sauce presented Sample 12 >2,400 in table form and described based on SNI Sample 13  >2,400 7388-2009 about the maximum microbial Sample 14  >2,400 limit on chili sauce for MPN Coliform 100/ml. And Regulation of the Source: Primary Data 202 Head of the Food and Drug Control Board of The results of coliform the Republic of Indonesia Number bacterial examination in Chili sauce HK.00.06.1.52.4011. About the maximum samples at food stalls around gorontalo limit of microbial and chemical contamination city snack market in lactose broth media in food with APM Coliform maximum 100/g. showed that there were 11 positive RESEARCH RESULTS samples with a percentage of 78.6% of the Microbiology tests on sambal 14 samples. The growth of bacteria in the conducted in the Microbiology Laboratory media lactose broth (LB) characterized by of Universitas Bina Mandiri Gorontalo the formation of gas and the formation of obtained the following results: noise, and 3 negative samples with a Coliform Bacteria Guessing Test presentase value of 21.4%. Preliminary On Lactose Broth Media (LB) analysis results on coliform guessing test were found positive results in a sample of Based on the research that has been 1 mpn 43/ml, 3-digit MPN 150/ml sample, done, the results and observations of the sample 4, 5, 6, 11, 12, 13, 14 MPN guessing test using lactose broth media number >2,400/ml, MPN number sample (LB) by looking at the absence of 8 4/ml and sample 10 MPN 15/ml number coliform bacteria characterized by the characterized by gas formation in durham onion of noise and the formation of gas in tubes and discoloration or noise in the media are presented in the following media lactose broth (LB). table: Coliform Bacterial Booster Test Table 1. On Eosin Methylen Blue Agar (EMBA) Coliform Bacteria Guessing Test Results Media In Lactose Broth Media (LB) Based on the research that has been done, the results of coliform bacterial Sample Guessing MPN booster test on Chili sauce using Eosin Code Test value Methylen Blue Agar (EMBA) media are (+) (-) presented in the following table:

14 Journal of Health, Technology, and Science (JHTS) Ana Pepiana E-ISSN: 2746-167X, Vol. 1, No. 2, December 2020

characterized by red gram coloring results. This corresponds to the

morphological characteristics of bacteria Table 2. Escherchia coli and Enterobacter Coliform Bacteria Booster Test Results aerogenes. in Eosin Methylen Blue Agar (EMBA) Media DISCUSSION Sample Chili sauce is a sauce of chilli base Colony Colors Percentage Count that resembles porridge and is usually added to other ingredients such as salt, Metallic Green 1 9 % shallots and garlic. Chili sauce has a taste Pink 10 91 % that varies according to the level of Total 11 100 % sasan. Chili sauce in its manufacture is Source: Primary Data 2020 still quite simple so that it can allow the The result of bacterial growth in the onion of pollution infood[14]. media Eosin Methylen Blue Coliform is a group of stem-shaped Agar (EMBA) was found 1 sample that gram-negative bacteria that generally had a metallic green colony with a produce gas if bred in lactose percentage of 9% which is the medium. The presence of Coliform in characteristic colony of fecal food indicates that it was once Coliform bacteria in this case suspected to contaminated by stools and it is likely that be Escherichia coli bacteria and 10 other it contained Escherchia coli,however, samples had pink and transparent colonies these results are not an absolute result with mucus which is a colony of nonfekal because non-pathogenic bacteria and not Coliform bacteria in this case suspected from the gut can also lead to positive enterobacter aerogenes bacteria. Coliform test results[ 13] . Pollution in food is the entry of Coliform Bacteria Supplement Test On unwanted foreign objects/materials Gram Staining contained in a food. Broadly, food Based on the research that has been contamination is divided into four types, done the results of coliform bacterial namely mechanical contamination, complementary tests on gram staining are biology, chemistry, and microbiological presented in the following table: contamination. This microbiological Table 3. contamination is one of which is that there Coliform Bacteria Supplement Test are coliform bacteria in food that can [10] Results on Gram Staining cause health problems in humans . Appearance on Coliform is a bacterium used as an Complementary 1,000x indicator of human or animal feces Test Magnification pollution and shows poor sanitation of Microscope food and drink. The arrival of enteropathogenic or toxic microbes that Cell Shape Stem are harmful to health. Coliform bacteria Nature Gram Negative are used as indicators of organisms because they are easy to find in a simple Source: Primary Data 2020 way. The absence of Coliform in food and The results of the gram coloring test drink indicates that the food and drink is on the sample examined in the microscope contaminated with bacteria and is the at 1000x magnification resulted in stem- cause of the healthdisorder diarrhoea[15] . shaped bacteria and gram-negative

15 Identification of Coliform Bacteria in Chili Sauce in Food Stalls around Gorontalo City Snack Market

Based on the results of research that drink means it is possible that the food or has been done on Chili sauce samples in drink contains Escherichia coli[15] . the food market around Gorontalo city Coliform bacterial pollution in Chili snack market conducted in the sauce can occur due to various factors, Microbiology Laboratory of Bina Mandiri namely, unhygienic equipment, Gorontalo University, from the results of processing process, as well as raw the examination in the guessing test materials and environmental showed that from the 14 samples sanitation. The market conditions of examined there were 11 samples with a Gorontalo city are located near each other percentage of 78.6% tainted with and are on the roadside, which can Coliform bacteria. In the results obtained potentially be polluted by pollution and there are 8 samples that have dust. Coliform bacteria in food indicate mpn coliform value >100/ml the arrival of toxic microbes that are which proves that the Chili sauce sample harmful to health. The higher the level does not meet the requirements specified of coliform bacterial contamination, the by SNI. Based on SNI 7388-2009 about higher the risk of the presence of other the maximum microbial limit on chili pathogenic bacteria, the presence of other sauce for MPN Coliform 100/ml. pathogenic bacteria, the presence of Coliform consists of 2 groups coliform is indicative ofinadequate namely fecal coliform group and nonfekal process or sanitation conditions[ 10] . coliform. In the test of 11 positive samples Most of the samples were positively and implanted suspension in emba media contaminated with coliform bacteria aseptic using inoculation needles obtained due to the poorly maintained condition of the results of 1 sample that had a greenish- food stalls and there were many flies red colony with metallic lightning was a around the stalls. Insects that become colony of fecal Coliform bacteria in this vectors of the spread of disease-causing case suspected Escherichia coli bacteria such as Escherichia coli (causing bacteria and 10 other samples had pink diarrhea) one of them is flies. Food stalls and transparent colonies with mucus being that are the site of some research are colonies of nonfectual Coliform bacteria. adjacent to trash cans that can affect or Fecal coliform bacteria are bacteria can be a source of contamination[11] . derived from human and animal Contamination in chili sauce can excrement. From the results found is a occur not only from environmental factors group of fecal coliform bacteria that or insects that become vectors of bacterial contaminate Chili sauce suspected to carriers but can also occur due to sellers be Escherichia coli bacteria. This is who in the presentation of food do not evidenced by the growth of bacteria in wash their hands using soap first. Dirty or petri dishes resulting in a metallic contaminated hands can be an greencolony. The metallic green colony intermediary for the transfer of bacteria or indicates that the pH in the other microorganisms into the food lacotose fermentor is quite low, while the served. Sellers who do not wash their pink colony indicates that the lack of hands using soap first can cause the acidity and enzymatic containing of possibility of bacteria in their hands gelatin is a source of nitrogen. Non- and contaminate the chili sauce[12]. fectual Coliform bacteria are bacteria Hand washing is an important thing derived from dead that workers should do if they are animals/plants. Coliform testing if it involved in food handling. Although it shows positive coliform results in food or appears to be a mild and often trivial

16 Journal of Health, Technology, and Science (JHTS) Ana Pepiana E-ISSN: 2746-167X, Vol. 1, No. 2, December 2020 activity, it has proven to be quite effective in preventing contamination of food. Shop CONCLUSION owners and workers usually only wash Based on the results of research that their hands using water, some even use has been conducted on chili sauce at food laundry water and only clean that is stalls around gorontalo city snack market visible to the eyes. This is in line with from all samples there are 11 positive previous research suggesting thatwhat can samples with a percentage of 78.6% affect food and beverage contamination containing coliform bacteria and 3 by bacteria is the cleanliness of the seller, negative samples with a percentage value the poor hygienic practices of merchants of 21.4%. A positive result was found in and the poor behavior and knowledge of [11] one sample of fecal Coliform bacteria the learner . suspected to be Escherchia colibacteria, Food ers in the snack market do not and 10 samples were suspected pay attention to hygiene and sanitation of nonfectual Coliform bacteria. when processing food, which is in accordance with the Decree of the REFERENCE Minister of Health of the Republic of [1] Adi, Febri Prasetyo. 2017. Indonesia No. 1096/Menkes/PER/VI/20- Undercover Science: Understanding 11. This is due to a lack of knowledge what Science Hides in Other from the seller about the hygiene Ways. : Diandra Kreatif. requirements of the behavior of the learner [2] Arisman. 2012.Nutrition in Life when processing food. Personal hygiene Cycle. EGC. : 193-195. when processing food is necessary to [3] Arnia., Citizen, produce food that avoids the fatal effects Efrida. 2013. Identification of of food poisoning and avoids coliform Coliform Bacterial Contamination In bacterial contamination. Fresh Beef SoldIn Markets Around As for some ways that can be done to Bandar Lampung City. MAJORITY prevent the contamination of Coliform (Medical Journal of Lampung bacteria, namely to maintain environ- University). ISSN 2337-3776. mental hygiene, not mix food waste with [4] Central Bureau of Statistics. 2018. raw food ingredients, cook chili sauce Number of Cases of HIV/AIDS, STI, until completely cooked, and in its DBD, Diarrhea, TB, and Malaria by presentation the seller is expected to District/City. Gorontalo: BPS. always wash hands first. [5] Chandra , 2012. Introduction to In addition there are several other Environmental Health. Jakarta: EGC ways to prevent contamination of E.coli Medical Book Publisher. bacteria in food, namely, Wash hands [6] Ministry of Health. 2010. Indonesia before and after preparing food and before Health Profile. Jakarta, Indonesia. meals, wash fruits and vegetables clean, [7] Depkes RI. 2015. Profil Kesehatan avoid cross contamination by using clean Indonesia. Jakarta. appliances, pans, and serving plates, keep [8] KBBI, Kamus Besar Bahasa raw meat away from other foods and from Indonesia. Arti Kata Warung other clean goods, only drink pasteurized Makan/Kantin. http://kbbi.web.id/kan dairy products (avoid raw milk, do not tin. Diakses Pada Tanggal 06 prepare food if you are attacked with November 2020. diarrhea, and ensure that all meat is [9] Kemenkes. Profil Kesehatan cooked properly and cooked Indonesia. Jakarta: Kementrian Kesehatan Republik Indonesi. 2018.

17 Identification of Coliform Bacteria in Chili Sauce in Food Stalls around Gorontalo City Snack Market

[10] Kuswiyanto. 2015. Bakteriologi Keguruan, Universitas Islam Negeri 1(Buku Ajar Analis Kesehatan). Raden Intan. Lampung. Jakarta: ECG. [13] Septiana, Rini dkk, 2015. Analisis [11] Mega Mirawati, dkk. Cemaran Bakteri Coliform Pada Saus 2014. Salmonella pada Jajanan yang Tomat Yang Dijual Di Wilayah Dijual di Warung dan Luar Warung Banjarmasin Utara Dengan Metode Most Sekolah Dasar. Jurnal Ilmu dan Probable Number (MPN). Tangerang: Teknologi Kesehatan. Vol. 1 No. 2. 2014. Akademi Farmasi ISFI. Hal 145. [14] Sutomo, Budi. (2014). Chili sauce & [12] Saadah Putri Farida. 2017. Analisis Saus. Jakarta: PT Kawan Pustaka. Bakteri Coliform Dalam Es Batu Dari [15] Wardani, Devi Listiana dan Setianingrum Berbagai Kantin Di Universitas Islam Zulia. 2019. Identifikasi Bakteri Negeri Raden Intan Lampung. Karya Escherichia Coli Pada Saus Makanan Tulis Ilmiah. Jurusan Pendidikan Jajanan Di Sekitaran Kampus Universitas Biologi, Fakultas Tarbiyah dan Muhammadiyah . Jurnal Kesehatan. 12 (2) 2019, 91-101.

18