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Contamination Level of Mold: Entero Bacteriaceae Family

Chapter III 


3.1. Introduction

The research methodology used a qualitative approach. The study has been conducted in November 2016 in Bandung city. The survey, observe and microbiology contaminant sampling was collected from food vendors premises as the study population. The study design used to evaluate knowledge, attitude, and practice (KAP) and contamination level of mold also Entero Bacteriaceae family. In addition, socio-demographic questions and others premise information that can be predicted factors for understanding the relationship between food vendors’ characters.  Furthermore, support from the local government as a regulatory role on food street activities was investigated to identify and to understand how to enhance better implementation in food hygiene.

3.2 Methodology Design and Study Instrument

3.2.1 Methodology design

The data survey presented the understanding, behavior, and implementation of hygiene practice by food vendor premises. The aspects were all assessed as follow:

  1. Knowledge; an assessment of hygiene practice knowledge was carried out using an interview survey.
  2. Attitude; Likert 5 points agree-disagree questions were asked for measuring the food handlers attitude.
  3. Practice; an observational study of food hygiene was used in the assessment. This audit used the score for measuring of its implementation.
  4. Microbiological contaminations sampling; an investigate study of cleanliness of cooking utensil.
  5. The presence of an implementation of regulatory aspects of street foods.

The social demographic questions and food premises information were also surveyed to explore the characteristics of respondents.


3.2.2 Study Instrument

In this present study, we used a constructed questionnaire based on the references. This questionnaire consist of (1) General questions regards to social demographic characteristics, (2) Food premises information (3) Knowledge, Attitude, Practice questions (4) The presence of an implementation of regulatory aspects.

Socio-demographic questionnaire as a complementary questionnaire consists of factors predicted that influence on respondent’s (food street vendors) knowledge, behavior and practice on food safety and hygiene which are constructed based on literature such as age, sex, marital status, education level, number of employment, employee type (self employer/ employee), how long they have been a seller and also their profit per day.

Food premises information showed the premises condition and type of food that sale, food preparation, food packaging material, type of facilities (i.e stall, mobile street vendors, tend, stand, or handed), and also hour length of sale.

The knowledge questionnaire, consists of 10 questions containing multiple choices questions, was giving to respondent for reviewing their awareness on food hygiene. The questions concerning knowledge about food hygiene from storage, cleanness, preparation and personal hygiene linked to transmission of microorganisms, and respondents were asked to choose one answer from among three options.

The questions of attitude consist of 10 questions. The answer options for these question used Likert scales ranging from 1 (very disagree) to 5 (very agree) to show a subjective response on their agreement on food hygiene implementation. The scores for attitude were similar with the range from 1-5, the higher score represents better manner on food hygiene.

The scores for practice on food hygiene were calculated from 50 questions based on an audit that observed on premises. The appearance of reducing cross contamination on food handlers/vendors was noted e.g. personal hygiene, food storage, pest control and utensil/ equipment. The source of water supply was investigated also the ice (whether commercially bought or self-made). The presence of food exterior facilities and sanitary utilities was determined e.g. availability of toilets, adequate washing facilities, sink, surface, waste, and food packaging. The availability of cool storage/refrigeration storage was also checked.

For microbial contaminant testing, VBRG dip slide type was used to determine whether there is evidence of entero bactericeae and mold in the food street premises e.g. cutting board, utensil, plate, glass and other food equipment.

3.3 DataCollection

An interview survey was carried out in one sub-district in Bandung city in November 2016. The minimum number of food street vendors was randomly selected as much as the result from prior validation questionnaire tests. The validation of questionnaire process has been conducted in July 2016 in Bandung city for improving the questionnaire and determining the appropriate sampling size also an area which should be chosen. The sample size minimum required for the study are calculated based on as the result of the prior validation study. Based on the level of significant of 5% with a confidence interval of 95%, 253 premises/ respondents are needed for this study. In this cross-sectional study, we surveyed 272 respondents from one sub-district.

Face to face interview was conducted in November 2016 by using Bahasa/Indonesian language and common Sundanese language as an ethnic language in the study area. All study instruments were in Bahasa/ Indonesia version and interview were taken on their premises. The study was supported by assistant researchers, as interviewers, who had food science background and understood both languages. The interviewer had been trained with regard to study such as food safety in a brief view, how to conduct the survey, interview method, study instrument, and ethical requirement including informed consent. The interviews were set in respondents’ premises to easier for answering the questions and to observe their practice on food hygiene. Informed consent form was read out by the interviewer. The interview taken as well as the respondents  were agreed to participate, questions were read out  by interviewers, and interviewers also marked the answer on the questionnaires instruments. The interviewers also acted as an auditor for observing the hygiene practice on their premises and recorded into practice audit sheet.

Sampling for determining microbial contamination used a dip slide that consists two agar sides, the first side the red side was used to confirm the presence of Entero Bacteriaceae family and the second side that flaxen color for mold. The sampling procedures were:

  1. Remove the dip-slide from the tube by pulling the plastic tab at the top of the dip-slide. Take care not to allow the dip-slide agar (the jelly type substance on each side) to hit the edge of the clear plastic tube.
  2. Place dip slide agar down onto the surface, and make sure the entire agar has full contact with the surface.  Keep the agar applied to the surface for approximately 20 seconds, then turn over to the other side and apply the other agar to the surface.
  3. Replace the dip-slide into the sterile plastic tube taking care not to damage the agar. Next, place the dip-slide tube into a dip slide incubator at 37 0C or warm place. The incubation period is 2- 3 days for bacteria. The optimum temperature for most mold/yeast is from 27 0C to 30 0C and the incubation time is 2 – 7 days.
  4. After 3 days, generally, both of results were read and recorded. Bacteria present in the sample liquid will grow and form colonies. The selective agar for Entero Bacteriaceae colonies will appear in red. A bacterial reference chart is used to determine the number of bacteria in the sample. The bacteria reference level has five charts that showed how many colony-forming-unit( CFU)/cm2 e.g. 2.5 (very slight growth), 12 (slight growth), 40 (moderate growth), 100(heavy growth) and 250 (very heavy growth). While the evidence of mold form appears in the furry colony. The chart of mold consists of slight (0.4), moderate (1.6) and heavy (4).

3.4 Statistical analysis

Food safety knowledge scores for the respondents were, calculated based on the multiple choice answer from  each question, only for the items of correct answers was assigned a score: +1 and 0 in the case of don’t know/wrong option.

For ten attitude questions were calculated from the Likert scale point 1-5, for question number 2, 6 and 9, the reversible point was used to calculate the point because it was a negative question.

The answers were classified as +1 point when the auditor/interview found out the correct evidence on respondent’s premises and 0 points when they failed to find it. However, not available marked for a respondent who did not have the correlation item with the type of food. The total percent score for the respondents’ attitude was then calculated by sum up all score in total.

Statistical analysis of the association between Knowledge, Attitude and Practice (KAP) questionnaire answers, social demographic characteristics and food information also microbial contamination level were carried out to determine the relationship between them and explore the pattern.

Data were analyzed by SPSS version 20. Based on the level of significant of 5% with a confidence interval of 95%, all analyses were considered statistically significant at P ≤ 0.05.

The descriptive analysis, Fisher’s χ square test, was performed to describe socio-demographics characteristics of the respondents. Analysis of variance (ANOVA) and t-test was used for comparing mean scores between all factors and observing the relationships between KAP and predicted factors that may influence on KAP.

Furthermore, to investigate whether this relationship systematically varied by specific socio-demographic characteristics for Independent variables included age, gender, and education level, and work experience, type of job which was full time or part time and having attended courses on food hygiene were conducted.


This study has approved by Ethics Committee of the University of Birmingham, School of Chemistry Engineering, United Kingdom. The identity of respondents will be treated securely and the outcome only will be used for academic purpose. The researcher and university supervisor will keep all data collection with confidential care.


Chapter IV


4.1 Sociodemographic

We obtained 272 responses from one area, 269 participants have completed the questions (response rate= 98.89%). Distribution of some socio-demographics characteristics respondents described in Table 1.

Variable category










Age group (years)




Young people (25-40)



Mature (41-60)



Elderly (> 60)



Marital Status







Education Level

No Schooling



Elementary school



Junior High School



High school









Profit per day (IDR)

< 50 K



50  – 100 K



101  – 200 K



201 – 300 K



301 – 400 K



> 401 K



Eat your own product




Rarely (1 – 4 times a month)



Sometimes (1-4 times a week)



Often (> 4 times a week)



Very often (every day)



Job type

Part time



Full time



4.2 Food premises information

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