International Journal of Microbiology and Allied Sciences (IJOMAS) ISSN: 2382-5537 February 2015, 1(3):19-24 IJOMAS, 2015 Original Research Article Page: 19-24 The Microbiological Quality of Ice Used to Cool Juices and Food Items in Lahore Asif Raheem 1 and Aftab Ahmad 2 1 Department of Microbiology and Molecular Genetics, University of the Punjab, Lahore, Pakistan 2 School of Biological Sciences, University of the Punjab, Lahore, Pakistan Abstract Corresponding Author: Aftab Ahmad School of Biological Sciences, University of the Punjab, Lahore, Pakistan Email: aftabac@yahoo.com Microbes are almost everywhere, including some of the items that we generally believe are free from germs. Openly available ice by street vendors and fresh juice shops in Lahore city were analyzed for their microbiological quality during the months of April to July, 2012.The total viable counts (aerobic colony forming units (CFU)) of all 10 samples from different location of Lahore were range from 8.8 10 2 to 1.9 10 5 cfu/ml. Almost all the sample collected from different places showed high microbial load and so unsatisfactory microbiological quality. It is concluded from results of the study that microbiology quality of ice was not good which could be due to use of bare hands, not cleaning of utensils and quality of water used to prepare ice. The use of such ice in food items also questions their quality so there is need of set proper standard on the quality of ice so the consumers could be protected from bad quality food and pathogenic microorganisms. Key words: Colony Forming Units, Ice, Pathogens, Street vendors, Sugarcane Juice. Introduction The World Health Organization (WHO) has defined street vended foods as, food and beverages prepared and/or sold by vendors in streets and other public places for immediate consumptions or consumption at later time without further processing or preparation [1]. The most important vehicle of foodborne illnesses throughout the world is contaminated food and drinking water [2]. According to many studies, it was illustrated that over the past decades in urban and rural areas of developing countries, foodborne illnesses causing high morbidity and mortality. Lack of awareness about the food safety, Poor and unhygienic conditions during food preparation are the major problems in spreading foodborne diseases in most developing countries. Because of inexpensive, readily available and different packages on food and juices offered by Street vendors, it is becoming popular among urban people as well [1]. The standard of street vender s food is very different in developed and developing countries so street food in developing countries is a big source of foodborne diseases. In most of developing countries, ice is sold by street vendors are widely consumed by millions of people. This is a readily available and affordable source of ice to many sector of population including urban poor, who cannot afford a refrigerator at home. They take ice block from 19
different ice factories outside city and bring those boxes to their vendors and place at wooden table near shops or on footpaths. Vehicles and other transport passing nearby and dust from road are a major source of contamination of ice. This ice is directly used into juices from fruits and vegetables and because of the fresh flavor, consumer prefer unpasteurized juice. So the final product is harbor of contamination because it is prepared by mechanical means, contaminated ice, punctured fruits and vegetables during harvesting and transporting from market to vendor. Raw materials and equipment use, traditional processing condition, improper handling, prevalence of unhygienic conditions all contributes to entry of pathogenic bacteria in juices prepared from combination of ice and fruits [3-5]. Even if there are no pathogenic bacteria but it often have high microbiological load. Lack of proper information and standards in these developing countries where street vended food is most prevalent is a major factor contributing to outbreaks of illnesses due to consumptions of unpasteurized fruits juice and openly available ice. There are several microbial studies on food in developed and developing countries reveals increase bacterial contamination of food with high risk pathogens to human health. In addition, many outbreaks were documented which are associated with the consumption of fresh and unpasteurized fruits juices which are prepared by vended ice [3]. In concerns with threats pose by bacterial pathogens in juice and other food material containing ice sold by vendors, the present work was designed to evaluate the microbial load or microbiological quality of vendor ice which is used in freshly prepared juice and other food products during April to July 2012 in different areas of Lahore, Pakistan. Materials and Methods Sample collection: Ten different ice stalls along with sugarcane squeezed juice machine, vendors or other fruit shops of Punjab University, Alama Iqbal Medical College, Jinnah hospital, Barkat Market and Moon Market, Lahore, Pakistan, were selected for sampling. Ice samples were collected in 50ml sterile falcon tube by wearing gloves and transported to the laboratory in an ice box for further processing. Additional information were also collected and recorded from shop owner and vendor about delivery of ice, processing and use in food items. Samples processing: Sample of ice were let to fully melt at room temperature and ice water was either used directly or dilution was prepared with autoclaved water. Bacteriological analysis was done through pour plate method. 10 ml of the sample was diluted with 90 ml of autoclaved water and mixed well (1-10 dilution). Serial dilutions were prepared and spread plate technique was used on enriched media (Luria-Bertani agar (L-agar)) Sigma- Aldarich. All plates were incubated under aerobic conditions at 37±1 C for 24 hrs. The mean number of colonies counted was expressed as log colony forming units (cfu)/ml. Results The samples were collected from 10 different locations of Lahore including barkat market ice store, hostel number 14 fruit shop (Punjab University), barkat market Bholi juice corner, Jinnah Hospital (sugarcane juice corner), barkat market (ice slush maker), hostel number 15 stop sugarcane juice vendor (canal road, Punjab University), Allama Iqbal Medical college fruit shop, hostel number 4 (Punjab University) canteen fruit shop, moon market ice store and moon market fruit shop. All the samples showed microbial load ranges from 8.8 10 2 to 1.9 10 5 cfu/ml. The maximum number of bacterial load in ice was recorded in barket market Bholi juice and Jinnah hospital sugar cane shop (6.52 10 3 ) followed by moon market ice store (3.62 10 3 ) and minimum number of bacterial load was at barket market ice store (8.8 10 2 ) and shops in University of the Punjab (Table. 1). The total numbers of colonies obtained with 1:10 dilution of ice water are shown in Figure 1 with lowest in IW-1 and highest in IW-3 and 4. 20
Sample ID Location Dilution factor No. of Colonies CFU/ml IW-1 Barkat market Ice Store 0 Numerous 10-1 88 8.8 10 2 10-2 80 8 10 3 10-3 70 7 10 4 IW-2 Hostel no.14 Fruit Shop 0 310 10-1 227 2.27 10 3 10-2 90 9 10 3 10-3 70 7 10 3 IW-3 Barkat Market Bholi Juice 0 Numerous Corner 10-1 652 6.52 10 3 10-2 225 2.25 10 4 10-3 150 1.5 10 5 IW-4 Jinnah Hospital (Sugarcane juice corner) 0 Numerous 10-1 652 6.52 10 3 10-2 270 2.7 10 4 10-3 190 1.9 10 5 IW-5 Barkart market (Ice Slush 0 Numerous Maker) 10-1 320 3.2 10 3 10-2 225 2.25 10 4 10-3 102 1.02 10 5 IW-6 Hostel no.15 stop Sugarcane Juice Vendor 0 Numerous 10-1 156 1.56 10 3 10-2 124 1.24 10 4 10-3 75 7.5 10 4 IW-7 Allama Iqbal Medical College Fruit Shop 0 Numerous 10-1 228 2.28 10 3 10-2 85 8.5 10 3 10-3 60 6 10 4 IW-8 Hostel no.4 Canteen Fruit Shop 0 Numerous 10-1 226 2.26 10 3 10-2 79 7.9 10 3 21
10-3 53 5.3 10 4 IW-9 Moon Market Ice Store 0 520 10-1 362 3.62 10 3 10-2 162 1.62 10 4 10-3 85 8.5 10 4 IW-10 Moon Market Fruit shop 0 Numerous 10-1 265 2.65 10 4 10-2 182 1.82 10 4 10-3 122 1.22 10 5 Table 1: Colony forming units (CFU) of all the samples collected from different places of Lahore. Figure 1: The total number of colonies from ice sample with 1:10 dilution Discussion Contaminated water is a big source of contamination of food worldwide and ice which is generally considered germ free can also be a source of contamination. In the present study, we checked the general hygiene of ice by aerobic CFU count of total aerobic microbial load in ice, which was in use to cool juices, drinking water and other food items in Lahore area. Lahore is 2 nd largest city of Pakistan by population. According to studies, deadly pathogens like, Listeria monocytogenes, E. coli, S. aureus and other intestinal pathogens involve in variety of diseases can be present in ice. In addition, Bacillus sp., Streptococcus sp., Staphylococcus epidermidis, Enterobacter aerogenes, Klebsiella sp., Citrobacter sp. and vibrio cholera were isolated and identified from contaminated ice [3]. In an outbreak study, Florida, USA, about 60 visitors were affected due to consumption of fresh non-pasteurized juice which was contaminated with salmonella [6] and one source of such contaminations in developing countries could be ice. According to a study conducted in Australia, about 427 confirmed cases of salmonellosis were reported in 1999 after drinking unpasteurized orange juice [3]. Another study illustrated that in India, an epidemic was reported in Pune city, which was because of sugarcane juice containing ice contaminated with Vibrio cholerae [7]. Numerous studies showed that microbial quality of manufactured ice is also compromised and could be source of contamination. Lateef et al., (2006) evaluated microbial safety of manufactured ice which was used to cool food and drinks in Nigeria and it was observed that microbial load of these manufactured ice samples were in the range of 1.88-3.20 10 4 cfu/ml, which was more than that of recommended microbial load for ice [8]. In our study, it was observed that quite high microbial load was present and especially at places which serve fresh juices to customers. Sometime people get food poisoning after drinking juice and beside many other factors (juice squeezing machine, operator etc.) ice could be a factor as it may contain a high dose of 22
pathogenic bacteria and that can cause food poisoning and other illnesses. As there are microbiological standard for drinking water, there should be for ice as ice is used in food items, juices and cold drinks almost everywhere in the world. The ice should be of microbiological quality and it should not have high load of microbes. The ice of satisfactory microbiology quality should have aerobic plate count or CFU that should not exceed 10 3 CFU/ml [9] but in our case just 2 samples (IW-2 and IW-9) had microbial count less than this, while all the other had high quantity of microbes and so was not of satisfactory level. Although we could only check the aerobic CFU, other factors are also necessary to evaluate like presence of total coliforms and E. coli, Enterococci, Salmonella and other related organisms, which is true indication of quality of ice and also have direct link with possible human diseases. This was a small study, with limited number of samples, a large scale and countrywide study is needed to have better idea about quality of ice in Pakistan. In addition, we observed that at all the places, the vendor added ice directly by hands so there was strong possibility of presence of above microbes along with ice as un-cleaned hand are great source of contamination [10]. Moreover, all the utensils which were in direct touch with ice were not very clean and vendors do not clean them quite often. In addition, ice was also not properly covered with plastic or other sheets to avoid direct contact of surrounding air. According to microbiological standards, the ice used in food items should be of same microbiological quality as potable water and E.coli and Enterococci should not be present in 100 ml [9]. The study shows that ice sold by vendors and used in fresh fruit juices was not of good microbiological quality as it contained high aerobic CFU/ml, so it also put question on the quality of food items in which it is used. The ice utensils in which ice is stored should be cleaned frequently and also should not be added into food items with un-cleaned bare hands. In addition, regulations should be set on ice quality and those standards should be followed all over the country to have good quality food for all the people. There is also need of public awareness that ice which is considered as germ free is not microbe free and it can contain potentially pathogenic bacteria which can put their health at risk. References 1. World health Organization (WHO). 1996. Essential safety requirement for streets vended foods (revised edition), food safety unit, division of food and nutrition, WHO, Geneva. 2. World health Organization (WHO). 1999. Food safety: an essential public health issue for the new millennium food safety program, department of protection of human environment, cluster of sustainable development and healthy environment, WHO, Geneva. 3. Victorian Government Department of Human Services, Food Safety Unit Melbourne, Victoria. 2005. Microbiological survey of freshly squeezed juices from retail businesses across Victoria. Available at: http://www.health.vic.gov.au/foodsafety (accessed September 12, 2014) 4. Oliveira ACG, Seixas ASS, Sousa CP, Souza CWO. 2006. Microbiological evaluation of sugarcane juice sold at street stands and juice handling conditions in São Carlos, São Paulo, Brazil. Cad. SaúdePública, Rio de Janeiro. 22(5):1111-1114. 5. Nicolas B, Razack BA, Yollande I, Aly S, Tidiane OCA, et al. 2007. Street-Vended Foods Improvement: Contamination Mechanisms and Application of Food Safety Objective Strategy: Critical Review. Pakistan Journal of Nutrition. 6(1):1-10. 6. Schmidt RH, Sims CA, Parish ME, Pao S, Ismail MA. 1997. A model HACCP plan for Small-Scale, Fresh-Squeezed (Not Pasteurized) Citrus Juice Operations. 7. Mosupye FM, von Holy A. 1999. Microbiological Quality and Safety of Ready- To-Eat Street- Vended Foods in 23
Johannesburg, South Africa. Journal for Food Protection. 1278-1284. 8. Lateef A, Oloke JK, Kana EB, Pacheco E. 2006. The Microbiological Quality of Ice Used to Cool Drinks and Foods in Ogbomoso Metropolis, Southwest, Nigeria. Internet Journal of Food Safety. 8:39-43. 9. Nichols G, Gillespie I, and de Louvois J. 2000. The microbiological quality of ice used to cool drinks and ready-to-eat food from retail and catering premises in the United Kingdom. Journal of Food Protection. 63(1):78 82. 10. Burton M, Cobb E, Donachie P, Judah G, Curtis, et al. 2011. The effect of handwashing with water or soap on bacterial contamination of hands. International Journal of Environmental Research and Public Health. 8(1):97 104. For Citation: Raheem A and Ahmad A. 2015. The Microbiological Quality of Ice Used to Cool Juices and Food Items in Lahore. International Journal of Microbiology and Allied Sciences. 1(3):19-24. 24