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Research-Protocol - Protocol
Medical Technology (BSMT1)
Emilio Aguinaldo College
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❑ Introduction/Background Background of the Study Street food trade is an ancient practice that remains in many developed and developing countries (Imathiu, 2017). The development of street food vendors is a result of urbanization, as the migration of people to urban districts led to the need to feed a vast number of workers (Canini et al., 2013). In many developing nations, street food vending is largely unregulated and unprotected by the government (Tonelada et al., 2018). According to Canini et al., (2013) most street food vendors can be found selling in busy streets; which are readily available outside of schools, churches, parks, and even in shopping malls (Buted & Ylagan, 2014). With the rate of urbanization in Dasmariñas accelerating, it comprises many neighborhoods and barangays where houses are placed next to one another; schools and open-air markets are frequently located nearby. Thus, providing to a huge number of Filipinos who lack the funds to establish a food service business to earn a living. Given the limited budgets, most students opt for affordable street foods that are easily accessible as an alternative to more expensive dining options (Tonelada et al., 2018). People who rely on such meals are frequently more concerned with its convenience than with its safety, quality, and cleanliness (Canini et al., 2013). However, it is claimed that street foods give rise to a significant threat to public health, increasing the likelihood of contracting foodborne illnesses (Tonelada et al., 2018). Salmonella, Listeria monocytogenes, enteropathogenic Escherichia coli are those examples of bacteria that causes foodborne disease, and street foods have been identified as a potential carrier (Campos et al., 2015).
The convenience and low prices of products may draw consumers' attention away from aspects of hygiene and sanitation. Some sellers and customers lack a thorough understanding of food-handling practices and the potential for foodborne illness. In fact, a recent report of Alarma (2022) revealed that approximately six (6) people were killed by the deadly diarrhea
outbreak resulting from contaminated street foods in Rasay and the public market in Toril District, Davao City, Philippines, the street food vendors care more about volume and price than freshness and cleanliness when purchasing raw foods to cook or sell (Imathiu,2017). Despite the serious public health and safety concern that many street-vended foods pose, poor knowledge among street vendors regarding basic food safety measures and inadequate public awareness of the hazards posed by certain foods have severely hampered the deployment of a precise scientific approach.
This study will focus on the detection of foodborne bacterial pathogens on street beverages and sauces and determine vendors’ food hygiene and safety practices. It aims to raise awareness about microbiological quality of street foods offered in selected areas of Dasmarinas, Cavite. In addition, look into vendors’ adherence to proper food hygiene and safety practices as well as assess if the street foods and beverages are safe for consumption. ❑ Aims of Study/Research Questions/Objectives/Hypotheses Objectives of the Study This study aims to investigate the presence or absence of bacterial isolates from street-vended sauces and beverages in randomly selected areas in Dasmariñas City, Cavite. It also intends to assess the street food handlers’ preparation and handling practices that serves as a predisposing factor to microbial contamination. Research Questions Several questions were addressed in the study:
- What is the average CFU/mL of bacteria in street food sauces and beverages isolated regardless of the location? a. Blood agar plate b. MacConkey agar plate
❑ Design/Methodology Research Design
The research design determines how data will be collected, analyzed, and used to answer the research question. In other words, it is the overall structure of a study (Grey, 2014).
This study will use a descriptive type of research. In accordance with Grove et al. (2013), descriptive design may be used to establish theory, define problems, make a judgment or determine what others in similar circumstances are doing (p). In particular, it can be used to make or formulate theories and support judgments with the information given.
With the use of comparative method, the researchers aim to describe and compare the microbiological quality of street foods sold in an enclosed area and open areas. Palamigs and street foods are among the most popular items sold by street vendors.
This study is entitled “Street Food Sauces and Beverages Microbiological Load and Vendors Food Hygiene and Safety Practices in Selected Areas in Dasmarinas City, Cavite, Philippines”. It will assess how the preparation methods of vendors selling street foods, inside the malls and outside where it is easily accessed affect their quality. The study will also identify the bacteria present in the samples. Furthermore, to relate how responsible the poor personal hygiene of the vendors is in the contamination of what they sell. Thus, helping to raise awareness for those who consume street foods regularly.
Sampling Design
Gathering data is necessary in order to understand the framework better. With this in mind, it is imperative that wise decisions be made regarding how and from whom data should be collected. The importance of this is emphasized by the fact that improperly collected data cannot be corrected through analysis (Etikan et al., 2016).
Purposive sampling will be used to identify food stalls where sample sauces and beverages will be collected in selected places of Dasmarinas City, Cavite. Etikan et al. (2016) also stated that in this technique, subjects are selected based on study purpose with the expectation that each participant will provide unique and rich information of value to the study. It is a non-random technique that does not require an underlying theory or a fixed number of participants. In other words, researchers determine what they need to know and they can find a participant who can and is willing to provide information based on their knowledge and experience. The purpose of qualitative research is to identify and select cases with rich information so that resources can be best utilized.
Additionally, researchers get to choose people or groups of people who are knowledgeable about their area of interest. It is also important in this approach to be accessible, ready to participate, and able to explain perspectives and experiences effectively.
Meanwhile, the respondents of the study will be asked to sign an informed consent form prior to collecting samples.
Locale of the Study
Study areas will include open and enclosed areas around markets, schools, and malls in Dasmariñas City, Cavite. A high number of vendors and consumers will be considered in choosing the location. There will be a total of 20 vendors participating in the study (10 vendors in open areas and 10 vendors in enclosed areas). The samples will be obtained in open areas within the vicinity of Area B Kadiwa (around Dasmariñas Integrated High School and Kadiwa Public Market), Paliparan 3 (around Paliparan Integrated High School and Paliparan 3 Market), Dasma Bayan (around Dasmariñas Elementary School and Francisco E. Barzaga Memorial School), Area E (around Dasmariñas
All samples collected from different food stalls will be inoculated into MacConkey and Blood agar plates. Inoculated plates will be incubated at 35-37 C for 18-24 hours. Bacterial growths will be checked after 18-24 hours. A bacterial colony counter will be utilized to identify the number of colonies present in the agar plates. The value of CFU/ml will be enumerated once the colony forming units (CFU) will be determined.
Bacterial Identification
To identify the possible bacterial species, the researchers will use the following procedures to determine the characteristics of bacteria that grew on the plates. ● Gram Stain A thin smear will be taken from the culture. The prepared smear will be fixed by flooding it with 95% methanol and letting it air dry. Smears will be overlaid with crystal violet for 30 seconds to 1 minute, and then will rinsed with distilled water. The smear will then be flooded with Gram’s iodine and rinsed with distilled water after 1 minute. Acetone or ethyl alcohol will be added drop by drop to the smear to decolorize the colors left after the addition of Crystal Violet and Gram’s Iodine. The smear will immediately rinse with distilled water. Safranin O will be added on the smear by flooding it. After 30 seconds, rinse the smear with distilled water then the slide will be allowed to drain and be dried. The smear will be examined under the oil immersion objective to view the characteristic reaction, morphology, white blood cells and other important structures. Gram positive organisms will stain blue-purple and gram negative organisms will stain pink-red. Biochemical Tests In this test, the physiology of bacteria will be determined according to species. The researchers will follow the standard protocol developed by Tille (2013). ● Triple Sugar Iron (TSI) Agar
Before use, the TSI medium will be warmed to room temperature. Through the use of a sterilized straight inoculation needle, a pure culture of the organism will be obtained and tested. Testing will be conducted on well-isolated colonies. Inoculation will be done by first stabbing through the center of the medium to the bottom of the tube and the needle will be used to streak the surface of the slant. The tubes will be incubated aerobically for 18-48 hours at 35-37 then observe the reaction. A red slant/ yellow butt (alkaline/ acid) indicates fermentation only. A yellow slant/ yellow butt (acid/ acid) indicates the fermentation of dextrose, lactose, and/ or sucrose. A red slant/ red butt (alkaline/ alkaline) indicated the absence of carbohydrate fermentation results. ● Lysine Iron Agar (LIA) Before use, the LIA medium will be warmed to room temperature. Through the use of a sterilized straight inoculation needle, a pure culture of the organism will be obtained and tested. Testing will be conducted on well-isolated colonies. Inoculation will be done by stabbing through the center of the medium to approximately within 3 mm of the bottom of the tube. A needle will be used to streak the surface of the slant. The tubes will be incubated aerobically for 18- hours at 35-37 C. The reaction in the medium will be observed. Positive results in Lysine Decarboxylation will have a purple slant and purple butt (alkaline). Negative results will be purple slant and yellow butt (acid). Lysine Deamination positive results will have a red slant and negative results will be the slant remains purple. H2s positive reaction will have a black precipitate and negative result has no black color development. Gas production will be demonstrated by the presence of bubbles or cracks in the medium. ● Sulfide, Indole, Motility (SIM) Before use, the SIM medium will be warmed to room temperature. Through the use of a sterilized straight inoculation needle, a pure culture of the organism will be obtained and tested. Testing will be conducted on well-isolated colonies Inoculation will be done by stabbing a half part of the bottom of the tube. The tubes will be incubated aerobically for 18-48 hours at 35-
Research-Protocol - Protocol
Course: Medical Technology (BSMT1)
University: Emilio Aguinaldo College
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