Bacterial and fungal burden of local packing papers used for roasted meat in Kano Nigeria

Background: The appetite for fast food in developing countries continue to rise although plans to regulate how these food are safely delivered to the consumers are not common if they exist. Objective: To determine the Bacterial and fungal burdens of local packing papers used for roasted meat in Kano State Nigeria. Materials and Methods: Standard microbiological methods were used to determine the bacterial and fungal loads of 108 locally improvised packaging papers for fast food roasted meats in strategic locations of the Kano metropolis. The packaging materials were obtained from consenting roasted meat vendors in the Kano metropolis. Appropriate aseptic precautions were adopted to ensure a good, reliable, and reproducible result. Result: The mean aerobic bacterial count observed ranged between 9.9 x 10 4 and 2.47 x 10 5 – 3.09 x 10 2 cfu/ml and 1, 24 to 3.09 x 10 2 cfu/ml for fungi respectively. High counts of both bacteria and fungi were found in samples of meat wrapping papers. B. cereus and E. coli were isolated from 55% and 100% of samples of meat wrapping papers respectively. The roasted meat wrapping papers included old newspapers (80%), used exercise books (8%), and disposed of office printed stationeries (12%). The most probable bacterial number MPN showed that the papers for meat packaging were more exposed to contamination (75MPN/ml) while the paper plate had (7MPN/ml). Conclusion: The papers used for roasted meat fast food were found to be unsafe and unfit for packaging purposes as they can easily be a source of disease epidemic due to the confirmed loads of bacterial and fungal pathogens. The factory sterilized paper plates are also not free from microbial contamination but might be due to exposure to unsanitary storage and marketing conditions during delivery to end-users.


INTRODUCTION
The spread of communicable diseases by personnel who handle ready-to-eat food and also the use of certain food packaging materials constitute a possible health hazard.This phenomenon appears more likely in third-world countries like Nigeria, where public health awareness is poor and where most food vendors are not certified medically by appropriate food handling authorities (1).One of the most serious and widespread risks from foods is the occurrence of potentially pathogenic microorganisms.The Food and Drug Administration (FDA, USA) and the National Agency for and Drug Administration and Control (NAFDAC-Nigeria), including state and local government health offices, offer strict requirements for the handling of foods to reduce microbial hazard leading to disease outbreaks (2).Mishandling of precooked, frozen, refrigerated, catered, or vended foods may give rise to contamination with potential pathogens, which can cause disease outbreaks.Contamination may come from unsanitary food distributors, poor health practices, such as failure to wash hands thoroughly or have open wounds.Mishandling is probably the most common problem in many food-borne disease outbreaks (1).
The basic materials for food packages and wrapping today include paper board, cellophane, steel, and aluminum.Aluminum foils, cellophane, and other papers meant for packaging are costly and beyond the reach of ordinary street vendors who serve the general public with irresistible roadside ready-to-eat fast food.This may explain why packaging and wrappings are done with improvised dilapidated old newspapers and exercise books destined for recycling.Another list of improvised packaging materials in the study area include but not limited to: Cement bags, disposable papers, old aluminum plates/cups, Conventional packages forms include wraps, bags, pouches, cartons, set-up boxes, cans, bottles, pails, drums, barrels, and bulk containers (3).These items though manufactured under strict aseptic conditions are later exposed in shops, supermarkets, and other retail outlets, and therefore may be liable to contamination.Food may be contaminated from utensil surfaces and pieces of equipment.Bacteria, including pathogenic organisms, might be left in food particles or moisture on perfectly washed crockery and utensils.It is unsafe to use contaminated utensils and containers particularly for cooked foods not intended for immediate use (4).Outbreaks of food poisonings frequently occur as a result of improper food preparation in which cross-contamination with inadequate storage or cooking was implicated in many instances (5).Dishcloth and sponges meant for cleaning eating utensils were recognized as a potential source of spreading micro-organism, and it was observed that bacteria persisted on these vehicles (6 and 7).
Micro-organisms have a wide distribution and are often found in a wide variety of habitats.A significant number of microbes can survive and use the air as a means of dispersal.There is a possibility of these microbes to be deposited on the eating materials such as paper plates or improvised packing materials such as newspapers, cement bags, milk bags, polythene bags, and many others.
In some instances molds may grow on such papers which are ready to use, raising the possibility of packaging mold and fungal spores together with the meat unintentionally.Since it may difficult to prevent or dissuade people from patronizing the roadside fast-food vendors that are likely to use these improvised packaging materials, one way the ignorant and innocent general populace can be assisted may be by determining how safe these packaging materials may be to design effective intervention against potential disease outbreaks.The above reason calls for an investigation into the possible microbial carriage pattern of such wrapping papers and disposable plates.This study was therefore designed to investigate the microbiological safety of disposable roasted meat wrapping papers and plates used in serving and preservation of fast food in homes and cafeterias in Kano.Thus, safety considerations in this study will entail determining bacterial and fungal load, detecting bacterial identities, and using this result to confirm the safety of different packing papers used to wrap meat in the Kano metropolis.

MATERIALS AND METHODS
A total of 108 pieces of paper plates and meat wrapping paper (representing a significant population of the meat vendors in the study area) were collected from three different locations in This open access publication is Licensed under a creative common's attribution 4.0 international License 3 each of the six metropolitan areas of Kano State.The samples comprised of 54 samples of each of locally improvised wrapping papers and 54 paper packaging plates respectively.Analyses of the Samples for bacterial isolation and count Inoculation of samples was carried out using swabrinse and standard plate count methods (8).Each of the samples was soaked in 100ml aliquots of sterile buffered (0.1% w/v) peptone water (Oxoid) for 20 minutes at ambient temperature.
The resultant washed water was used to carry out serial dilution up to the fifth dilution (2).One milliliter of each dilution was taken using a sterile syringe and placed into each of the appropriately marked duplicate dishes of 15ml PCA (kept at 45 0 C in a water bath).Within 15 minutes of the time of original dilution, media were poured into each of the Petri dishes, sample dilution, and agar medium was thoroughly and uniformly mixed and allowed to solidify.The prepared media were inverted and incubated at 30 0 C for 72hrs.Three tubes each containing 9ml MacConkey broth with an inverted Durham tube were each inoculated with 1.0ml of the sample washed in sterile physiological saline to give a 1 in 10 dilutions.These were repeated with 1 in10 2 and 1in10 3 dilutions.All tubes were incubated at 37 0 C for 24hrs (2).Tubes that show gas production was recorded.A loopful of broth from each gas positive test tube and the presumptive test tube was inoculated into a separate tube containing brilliant green lactose bile broth (BGLB) and the tubes incubated at 37 0 C for 48hours.The formation of gas confirmed the presence of coliform bacteria.The numbers of positive tubes were recorded and the most probable number of the coliforms determined from and most probable number table (2) Simultaneously with the confirmatory test, transfer was made from positive presumptive tubes to other tubes containing enrichment medium, and the tubes incubated at 37 0 C for 24hours.Gas and acid formation was recorded and the bacteria density was estimated from the most probable number table.Gas and acid production confirmed the presence of fecal coliform (2).These tests were carried out by the procedure described by Odoki et al, (9).A loopful from each positive tube from the presumptive test was transferred to a spate tube of E. coli broth reincubated at 37 0 C for 48hours.Gas and acid production were recorded.Levine's eosin methylene blue agar (L-EMB) plates were incubated from each positive EC tube and incubated at 35 0 C for 18-24hrs.The production of dark purple colonies with metallic sheen confirmed the presence of E. coli.Gram staining of the cultures was carried out and the presence of E. coli was confirmed by IMViC test (2).The procedure adopted from FAO (2) and Jean (10) 4.
Although we did not investigate what fuels the appetite for such roadside roasted meat, with potential health risk, it appears cost, ready-to-eat nature of the food, availability when needed and poor time management by customers may be playing a significant role in the patronage of roadside meat vendors.This is different when it is compared to expensive, impatience to procedural hygiene protocol for food preparation common at big hotels and restaurants, the need to place a food order and wait for a specified time before a portion of such meat is served, thus making people patronize the fast food vendors more than the standard food centers.Again, the handlers and the poor aseptic environment where such meat is packaged are also issues to be considered in understanding why the observed microbial burden exists in the first instance.Future studies should be able to outline the microbial burden of the wrapping papers visà-vis the microbial burden of the food handlers themselves to be able to design and implement an effective intervention during the associated epidemic (11)(12)(13).
High bacterial counts obtained in this study also corroborates with the work of Aboloma ( 14

Table 4 : Mean Fungal Count for + Standard Deviation for Paper for Meat Wrapping
2

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Aspergillus Flavius are increasingly prevalent as an opportunistic agent of tropical infection.Aflatoxin which are mycotoxins produced by such strains is undesirable in foods because they are potential health hazards and cause of epidemic and great mortality due to disease outbreaks in resource-limited settings.In conclusion: The papers used for roasted meant fast food were found to be unsafe and unfit for packaging purposes as they can easily be a source of disease caused by bacterial and fungal agents of diseases epidemic.Improved diagnostic and identification methods to improve recovery of isolates, appropriate sterilization process, and associated awareness campaign to prevent associated disease epidemic are highly recommended healthy females in Ekpoma Nigeria.OJHAS, 2007, 1: 2. 14.Aboloma RI.Microbiological Analysis of Bread Samples from Bakery to Sale Points in Ado Ekiti, Ekiti State, Nigeria.Biological and Environ Sc J for the Tropics 2008; (3):77-81.15.Fansami GO, Olukole SG, Kehinde OO.Microbial Studies of Table Scrapings from Meat Stalls in Ibadan Metropolis, Nigeria Implication on Meat Hygiene.African Journal of Biotechnology 2009; 9 (21): 3158 -3162.http://www.academicjournals.Org/AHB 16.Nwachukwu E, Ezeama CF, Ezeanyos BN.Microbiology of polyethylene -packaged sliced watermelon (Citrullus lanatus) sold by street vendors in Nigeria African J of Microbiol Research: 2008: 192 -195.17.Adegunloye DV, Agarry OO, Adedolu TT, Adetuyi FC.Effect of leaf-packaging on the microbiological assessment of some food items.African J of Biotech 2006: 5 (5): 445 -447.18. Adams MR, Moss MO.Food Microbiology.The Royal society of Cambridge.1995; 106 -110.19.Okegbue RN.Studies on the Microbiology of cassava flour.Nig.Food journal 1991: 9(85)-91