Microbiology Research 2019; volume 10:7463

Evaluation of microbial quality improper handling and use of tarnished of street in Karachi City, utensils.1 As defined by the US Division of Correspondence: Aneela Mehboob, Food Health and Human Services website, Research Group, Laboratory #115, Pakistan: An epidemiological poisoning also referred to as foodborne ill- Department of Microbiology, University of study ness or foodborne disease is any sickness Karachi, Karachi, Pakistan. that results from consuming food contami- E-mail: [email protected] nated with microbes or their toxins. Many Aneela Mehboob, Tanveer Abbas Acknowledgments: Dr. Aneela Mehboob of such foodborne diseases are known to desires to express her gratefulness and sincere Food Safety Research Group, cause serious health related disturbances.6 Laboratory #115, Department of gratitude to her respected supervisor Dr. Health issues are the real concern with Tanveer Abbas, Assistant Professor, Microbiology, University of Karachi, respect to street vended food. The way the Department of Microbiology, University of Karachi, Pakistan food is prepared, processed and presented Karachi, for his kind cooperation, inspiration, consequently affects its microbiological and valuable advice. quality.7,8 Adulteration can be introduced in food through several courses; nature of Key words: Foodborne pathogens; Street Abstract water utilized for is one of the ; Karachi; Pakistan. major causes contributing to food poison- The purpose of this research was to ing.9 Nonetheless, being the essential feed- Contributions: the authors contributed equally. evaluate the microbial quality of famous ing source of large number of people street foods vended in Karachi city and find Conflict of interest: the authors declare no including travelers, labors, shopkeepers and out the potential sources implicated in food potential conflict of interest. students at reasonably low prices and easy contamination. To proceed with this study, access, the importance of cannot dry and wet procedures were used via Funding: none. be questioned.10 In recent years major cities closed system questionnaire and gold stan- of Pakistan have suffered significant raise in Received for publication: 10 November 2017. dard techniques respectively. According to urbanization largely due to migrations from Revision received: 1 April 2018. the questionnaire survey, majority of people only rural areas. This pressure of increased pop- Accepted for publication: 9 April 2018. have basic knowledge of food borne sick- ulation has caused street food vending busi- ness i.e. 67% and health risks associated ness to grow extensively as these food This work is licensed under a Creative with poor habits i.e. 87%, nonethe- Commons Attribution NonCommercial 4.0 hawkers earn their livelihood by catering to less most individuals do not mind their License (CC BY-NC 4.0). the needs of large number of migrantuse work- cleanliness before eating on spot. A large ers far away from their home places.11 number of people had come across different ©Copyright A. Mehboob and T. Abbas, 2019 There are number of culinary dishes which Licensee PAGEPress, Italy food borne infections subsequent to eating are quite popular amongst the inhabitants of Microbiology Research 2019; 10:7463 street foods. Total mean aerobic count was Karachi city including , , doi:10.4081/mr.2019.7463 7.18±1.26 CFU/ml that was not under the Nehari, Barbeque items and Junk foods. acceptable microbial limits. The highest Mobile food sellers are the real public 20-23 total viable count 9.15±0.01 CFU/ml and eases throughout the world. Careless health safety concern as they prepare and the lowest total viable count 4.67±4.04 attitude of food vendors results in bacterial dispense their finished products in carts and CFU/ml recorded were of grilled chicken pathogens becoming more stubborn and vans where they are bound to face difficul- and fried fish respectively. Poor post han- resistant to numerous antibiotics. Many ties like unavailability of clean water, infe- dling and personal cleanliness of food hospitalization cases are associated with rior grade raw materials, lack of appropriate venders were ascribed to high bacterial foodborne diseases therefore treatments equipment and layouts, poor sanitation con- count in completely cooked street foods. turn out to be more challenging and frustrat- ditions, improper food processing and pack- ing.24-27 Therefore the aim of this study was aging techniques and absence of suitable to find out various food handling factors on storage facilities and waste disposal street that might lead to causing bacterial places.4,5,12,13 Correspondingly imprudent Introduction Non-commercial diseases to the users. For this various fac- treatment of such foods is the chief source tors have been assessed like personal Street food can be described as the food of food contamination. Food is an excellent hygiene of hawkers and vendors, and intrin- consumed or eaten on spot or as takeaway.1 medium for microbes to flourish and 9 sic and extrinsic environment (pH, temper- Street food is quite common and well expand their number quickly. Ready to eat ature and humidity), which ultimately affect accepted all around the globe. Different food especially made up of protein sources the quality of foods. parts of the world are represented by their such as poultry and provides a poten- typical street foods that are not only palat- tial vehicle for food borne microorgan- able and presentable but also reasonably isms.5,14-16 It was reported that food prod- priced thereby attracting local inhabitants ucts from animal origin and fresh produce Materials and Methods and tourists.2-5 Besides, street food is from plants like play a crucial role in enjoyed by all age groups and classes of spreading antimicrobial resistant bacteria. Selection of study site and people. Street food is ready to consume Roughly 10 to 20% of food borne illness are questionnaire design food and is promptly served while both raw caused by food handlers either by mishan- Karachi city is the most populous city in and cooked items are catered to the buyers. dling of food or their poor personal Pakistan and the 6th most populous metro- Health risks are more associated with the hygiene.17-19 Similarly noncompliance with politan city in the world. Karachi is divided raw foods, especially vegetables, as these the requirements of Good Hygiene Practice into six districts namely East, West, Central, inherently harbor high number of microor- (GHP) and Good Sanitation Practice (GSP) South, Malir and Korangi. North ganisms which are further worsened by is the main reason behind food borne dis- Nazimabad town and Gulberg town areas of

[Microbiology Research 2019; 10:7463] [page 1] Article

Central district Karachi were taken under number of bacterial count. From each dilu- broth and Mossel’s broth were used. consideration for research purpose (Figure tion, six drops of 20 µl were positioned on Subsequent to selective enrichment, inocu- 1). Closed ended questionnaire was Mueller Hinton agar plates. Inoculated plates lation was done from Lactose and Mossel’s designed to analyze the knowledge of pub- were left undisturbed until the drops were broth to selective media including lic regarding food related health risks. properly absorbed. Then plates were incubat- MacConkey’s agar, Salmonella Shigella Survey was comprised of basic questions ed overnight at 37ºC. Colonies were counted agar, Deoxycholate Citrate agar, and Eosin including awareness about foodborne dis- and tabulated after incubation period. Methylene Blue agar. Discrete colonies on eases and incidences of diarrhea, dysentery, selective media after incubation were hepatitis after consuming street foods. Enrichment and identification of inspected macroscopically and microscopi- bacteria cally then biochemical tests were performed Sample collection Enrichment technique was used for the for further identification according to 29 For the research, 54 ready to eat food isolation of bacteria belonging to Bergey’s manual of determinative biology samples were collected from local stalls, Enterobacteriacae family for which Lactose as shown in Table 1. shops and street vending sites at different locations in Karachi city from April 2015 to August 2016. Some food samples were pro- ceeded on time as soon as the sample was collected while the others were examined after 10-12 hours. To avoid any exogenous contamination, food samples were asepti- cally collected, maintained and stored at 4ºC in refrigerator until analyzed.

Isolation and enumeration of total viable count only 1 gm/mL of food sample was homoge- nized in 99ml of maximum recovery diluent and the beaker was let to stand for 15-20 minutes. Ten folds serial dilutions were pre- use pared by adding 1 mL homogenized sample in 9 ml blank (distilled water) up to 10-6 dilu- tion. From the last three dilutions (10-4, 10-5, -6 28 Figure 1. Map of Central district Karachi highlighted the research areas. Source: Official 10 ), Miles and Misra technique was per- web portal of Karachi Metropolitan Corporation (http://www.kmc.gos.pk/). formed in triplicate to determine the total Table 1. Biochemical characteristics of bacteria recovered from street foods. Biochemical E.aerogenes S.fonticola E.coli S.liquefaciens C.freundii S.grimesii Pseudomonas spp S.fecalis S.aureus L.adecarboxytaca tests IND - - + - - - - + - + MR - + + + + + + - - + VP + ------+ - - CIT + + - + + + + - - - NIT + + + - + + + - + + TSI A/Ag+ A/Ag Non-commercial A/Ag+ K/A K/Kg H2S A/A K/Kg A/K A/A A/Ag+ SIM --+ - + - - + - - + - + - + - - + - - + - + + Oxi/Cat -/+ -/+ -/+ -/+ -/+ -/+ +/+ -/+ -/+ -/+ URE ------Glu AG AG AG A AG AG A A A AG Lac A A AG - AG - a A A AG Suc A AG A - a AG a A A AG Mal A AG AG A AG AG a A A AG Mani A AG AG - AG AG - A A AG Man A AG AG A AG AG - A A AG Xyl A AG AG A - AG a a A AG Sor A AG AG - AG AG - A A - Raf A AG - - - AG - - - AG Dul - AG AG ------AG Fru A AG AG - AG AG a A A AG (-), negative; (+), positive; AG, acid and gas production; A, acid production only; a, weak acid production.

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Statistical analysis Pseudomonas spp 20% and Bacillus spp S. fonticola, S. liquefaciens, S. grimesii con- All experimental analyses were done in 18% were predominant followed by S. sidered as opportunistic organisms were triplicates and interpretations were present- aureus 6%, S. epidermidis 6% and S. fecalis also isolated from food samples. ed as Mean ± SD by Minitab software ver- 2% as shown in Figure 5. Among sion 17. Enterobacteriacae, E. coli 14% and E. aero- Total viable count genes 10%, both notoriously known to The mean bacterial count of the isolates cause food poisoning, diarrhea and dysen- was expressed as log CFU/gm (mL). Food tery cases, were also isolated. Similarly, was classified as acceptable if the bacterial Results Epidemiological survey Table 2. Evaluation of basic knowledge of public through closed system questionnaire Data with respect to the food and health based survey. awareness was gathered through structured Parameters Frequency n=70 based survey from public (Table 2). Survey was based on basic inquiries regarding indi- Age (years) viduals’ dietary patterns, cleanliness prac- <20 4 (5.7%) tices and awareness on food related health 20-29 42 (60%) risks. According to the survey, delicious- 30-39 6 (8.5%) 40-49 4 (5.7%) ness 68.57% and convenience 42.85% are 50-59 1 (1.4%) the primary explanations for the street food 60-69 1 (1.4%) popularity. These foods are so scrumptious Gender that individuals cannot get enough of it and Male 27 (38.5%) due to this reason people tend to eat out 3-4 Female 43 (61.4%) times i.e. 64% in a week. As indicated by Knowledge about food borne diseases the collected information, eating street Yes only 47 (67%) foods is more prevalent in the age group of No 10 (14%) 20-29 years and majority of the food lovers Kind of 13 (19%) are female i.e. 61.4%. According to the sur- Awareness regarding the major health risks with poor eating habits vey, 54% of the public prefer to eat on spot Yes 61 (87%) and 89.4% of these people do not wash or No use 9 (13%) sanitize hands before having a . This Preference of eating study showed that 46% of the general pop- On spot 38 (54%) ulation encountered malaise (nausea, vom- Takeaway 36 (74%) iting, and stomach ache) after devouring If on spot, do you wash your hands before eating? street food in the past. It was likewise Yes 36 (51.4%) noticed that 14% of participants do not have No 34 (49%) the information about foodborne diseases Junk foods make you eat more? and in addition, 13% of the public do not comprehend the significant health hazard Agree 48 (69%) related with poor dietary habits. Ready-to- Disagree 22 (31%) eat foods samples were collected from vari- Are junk foods healthy? ous street vending sites at different loca- Yes 4 (5.7%) tions of Karachi city. The most favorite No 66 (94%) food items to which public are attracted are How many times do you eat out a week? fries 71.4%, Barbeque items 45.71%, None 2 (3%) 34.28%, Nehari 27.14% followed by Fried 1-2 45 (64%) chicken 15% as shown in FigureNon-commercial 2. The 3-4 11 (16%) types of street foods analyzed in this study 5-6 4 (5.7%) were Nehari 10%, Haleem 10%, biryani 7-8 1 (1.4%) 10%, Barbeque items 10%, 2%, Fish >8 4 (5.7%) 4% etc. as presented in Figure 3. Out of 70 Instant nausea, vomiting, stomach ache after consuming street foods? volunteers, 29 contracted diarrhea, 25 Yes 12 (17%) reported to have suffered food poisoning, No 28 (40%) Sometimes 32 (46%) 10 experienced constipation, while 7 of them faced dysentery after consuming street Reason for consuming street food food in past as shown in Figure 4. Convenient 30 (42.85%) Delicious 48 (68.57%) Pleasant appearance 11(15.71%) Microbial quality of street foods Peer influence 6 (8.57%) An aggregate of 54 food items were Healthy 6(8.57%) microbiologically examined and all samples Habit 12 (17.14%) were positive for microbial growth. Both Discount 2 (2.85%) Gram-positive and Gram-negative bacteria Nice odor 12 (17.14%) were recovered from the food samples Inexpensive 15 (21.42%) (Table 3). Of the bacteria isolated, Curiosity 1 (1.42%)

[Microbiology Research 2019; 10:7463] [page 3] Article count was less than or equal to 5 log Table 3. Quantity of food items analyzed and microbes recovered from famous street foods in Karachi. CFU/gm (mL).30-40 The cumulative aerobic count of all tested street food samples was Street Foods Bacteria isolated 7.18±1.26 CFU/gm (Table 4). TVC of Nehari (n=5) E. coli, S. fonticola, S. aureus grilled chicken 9.15±0.01 CFU/gm was the highest and TVC for fried fish 4.46±3.86 Biryani (n=5) S. grimesii, P. cholaraphis, E. adecarboxytaca, S. fonticola, E.coli CFU/gm was the lowest among all food Korma (n=5) E. coli, E. aerogenes, samples. Tested food items were not found Haleem (n=5) S. liquefaciens Bacillus spp, gram negative rods to be within the acceptable microbial quali- BBQ items (n=5) Pseudomonas spp Bacillus spp ty limits and were potential source of food- items (n=4) Pseudomonas spp E. coli, E. aerogenes, borne diseases. The presence of high bacte- rial load in tested samples was attributed to Salad (n=1) Bacillus spp post handling like serving with bare (n=4) C. freundii, Pseudomonas spp, E. coli, S. fonticola hands and use of untidy utensils. Table 5 Finger fish & sauce (n=1) S. aureus, S. epidermidis, Bacillus spp demonstrated that how the tested street (n=4) E. coli, Bacillus spp, E. aerogenes foods are typically prepared, cooked, han- Pani (n=1) E. coli, E. aerogenes dled, and served. (n=2) Pseudomonas spp, gram negative rods items Grilled chicken (n=1) S. fonticola, S. epidermidis, C. freundii Discussion Stuffed chicken (n=1) Pseudomonas spp, gram negative rods Epidemiological studies are of great Mayo garlic roll (n=1) S. fecalis, C. freundii importance in researching the links between Chicken roll (n=1) E. aerogenes foodborne sickness and its causes. Being an Chicken Manchurian (n=1) S. fecalis ignored area of study in a developing coun- Chicken (n=1) Pseudomonas spp only try like Pakistan, there is no significant epi- demiological data to infer relationship Chicken roll (n=1) S. fonticola, P. flouresence between consumption of different street Wonton (n=1) Pseudomonas spp, C. freundii foods and the resultant foodborne diseases. Zinger burger (n=4) Bacilluuses spp, S. fonticola, Pseudomonas spp Several food poisoning outbreaks due to street foods were reported in from Table 4. Total bacterial counts [Mean log10 CFU/g (ml) ± standard deviation for foods 1983 to 1992 in Shandong province in with at least 2 samples & log10 CFU/g (ml) for 1 food sample]. which street foods were found to cause 691 Food items Mean log CFU/g (ml) SD (±) food poisoning cases and 49 resulting 30 deaths. Other studies also supported the Korma 7.63 – TNTC 0.73 high risk of consuming street foods culmi- Haleem 7.42– TNTC 0.61 nating in foodborne diseases.31-34 It was Samosa 8.43 - TNTC 0.03 reported in previous studies that more or less similar food borne pathogens including Zinger 7.33 0.90 E. coli, S. aureus, S. fecalis, S. typhi and Sandwich 6.80– TNTC 0.62 Bacillus spp were isolated from various Biryani 7.18 – TNTC 0.25 35-39 street foods. Nehari 7.49 - TNTC 0.52 The extrinsic and intrinsic environment Pani puri TNTC - under which the street foods prepared, processed and sold by vendors and hawkers Fish 4.46 3.86 were not suitable and did not fulfill theNon-commercial stan- Fish sauce 4.67 4.04 dards of GHP and GSP.40-42 pH of all the Salad 4.67 4.04 food samples and the temperature of envi- Grilled chicken 9.15 0.01 ronment where foods were prepared, exam- Fries 7.60 - TNTC 0.18 ined in this research were more or less sim- ilar and respectively classified under the Shashlik 7.0 0.0 range in between (6.5-7.0) and (30-37°C). Chicken Manchurian 7.02 0.35 This investigation described the impact of Wonton 7.31 0.15 seasonal variation on the type of bacteria Stuffed chicken 8.61 0.03 recovered from street vended foods. As Mayo garlic roll 6.79 0.17 sample collection was performed between Tikka roll 7.44 0.08 the periods April 2015 to August 2016. It was observed that the food items acquired Chicken roll TNTC - and handled in winter season had lower Dhaga kabab 6.99 0.30 bacterial count showing decrease in the Bihari boti 8.68 0.09 total aerobic count of microorganisms while Tikka TNTC - marked intensification in TVC was noticed Creamy tikka 8.21 0.02 in summer season. Correspondingly, from November to February, only gram positive Total 7.18 1.26

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Figure 3. Frequency of food items analyzed.

Figure 2. Main streams foods of Karachi.

Table 5. Street foods encountered in Central district Karachi with a description of the major components. Street foods Ingredients Nature of food Handling after cooking Source of contamination Nehari Meat, onion, spices Entirely cooked Served hot with spoon Equipment, eating bowl Wonton Minced chicken, onion, Deep fried, Served with hands or tong Hands spring onion, egg, spices dough, oil only Biryani Chicken/beef, rice, Entirely cooked Served hot with spoon Equipment, eating bowl spices, potato Chicken shashlik Chicken, onion, tomato, Pan fried Served hot Equipment, eating bowl capsicum, lemon, yogurt, spices use Grilled chicken Chicken, spices, Grilled Served hot with spoon Under cooked butter yogurt Stuffed chicken Chicken, vegetables, Deep fried Served with spoon mayo, cream, spices, egg Pani puri Flour, chana, onion, Mixture of fried, cooked Served with hands Hands, serving bowls, spices, potato, and raw ingredients; snack Haleem Beef, onion, spices, Served hot with spoon Equipment, eating bowl pulses, oil, wheat, rice Chicken Boneless chicken, tomato Cooked Served hot with spoon Equipment, eating bowl Manchurian sauce, onion, sugar, pineapple juice, spices, oil, soya sauce Chicken mayo Boneless chicken, Fried, cooked Served with hands Hands garlic roll mayo sauce, flour, spices, yogurt Non-commercial Sandwich Beef/chicken patty, egg, Cooked Served with hands Hands oil, tomato, onion, cucumber, ketchup, mint chitin, spices French fries Potato, spices Deep fried Served with hands Hands BBQ items Chicken/ beef, spices, Cooked on coal Served hot with hands Hands oil, butter, cream, onion, yogurt Zinger Chicken, flour, pepper, Deep fried Served hot with hands Hands chili sauce, ketchup, mayonnaise Samosa Boiled mashed potato, Deep fried, snack Served with hands Hands spices, dough Coleslaw salad Cabbage, spring onion, No cooking Served with spoon Fresh vegetables, carrot mayonnaise, hands and equipment pepper, lemon, sugar Finger fish Fish, spices, egg wash, Deep fried Served with hands Prolong storage, hands bread crumbs

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Figure 4. Incidence of food borne diseases consuming street vend- Figure 5. Percentage dominance of microbes recovered from ed food in Karachi. street vended foods in Karachi.

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Non-commercial

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