2014, World of Researches Publication Am. J. Life. Sci. Res. Vol. 2, Issue 3, 379-383, 2014 American Journal of Life Science Researches www.worldofresearches.com ORIGINAL ARTICLE Received 12 Jan. 2014 Accepted 14 Jun. 2014 Mycological Quality of Some Commercial Body Creams at Eke Awka Market, Awka Nigeria S.C Onuorah*, C.E Onugbolu, C.U Nwaoha, U.C. Okafor, I.E Obika and M.U Orji Department of Applied Microbiology and Brewing Nnamdi Azikiw, e University,P.M.B. 5025,Awka, Nigeria *Corresponding Author: S_Onuorah484@yahoo.com Abstract: A study on some commercial body creams sold at Eke Awka Market, Awka, Nigeria was conducted to isolate, characterize and identify the fungi present in them. Samples of five different brands of unused body creams were used in the study, with sabouraud dextrose agar as the growth medium and streaking as the method of cultivation. The fungi were isolated, characterized and identified on the basis of their cultural and microscopic characteristics as Penicillium sp, Fusarium sp, Mucor sp, Rhizopus sp and Aspergillus sp. The fungal counts ranged between10 cfu/g and 22cfu/g. Sample 4 had the highest count while sample 3 had the lowest count. Mucor sp and Aspergillus sp occurred most frequently in the samples while fusarium sp and Rhizopus sp had the least occurrence. The contamination of these body creams by the isolates may have come from a variety of sources such as the raw materials, air, soil, water, the producers and packaging materials. These isolates are known to cause diseases of man and animals and their presence in the body creams is a public health concern. This calls for an amendment in the existing formulation protocol and a strict adoption of good manufacturing practices to eliminate the entry and proliferation of these organisms. Keywords: Mycological, Quality, Commercial, Body Creams, Eke Awka, Nigeria INTRODUCTION Cosmetics are products that are intended to be applied to various surfaces of the human body to cleanse, perfume, protect, beautify, change appearance or correct body odour. They are mainly applied to the skin or hair and are not intended to permanently alter the physiology of the target organ, although some help with conditions such as dandruff, spots and poor gum health. The microbiology of cosmetics is therefore complex due to the wide range of formulations, manufacturing procedures and conditions of use 1. Most cosmetic products are based on water/oil or oil/water emulsions and contain variable amounts of nutrients. The raw materials used in cosmetic products include water, acids, alkalis, salts, oils, waxes, alcohol, esters, surfactants, emulsifiers, talc, clay, proteins, starches, botanicals, gums, resins, colour, preservative, antioxidants, chelating agents, fragrances and pigments 2 379
Onuorah et al., 2014 that make them efficacious and sufficiently elegant to be acceptable to users. Some of these ingredients may serve as good sources of nutrients, carbon and energy for the growth and proliferation of molds in the products resulting in their contamination and spoilage 2. Microbial contamination is one of the major causes for cosmetics recalls. Cosmetics are subject to variable and constant microbial contamination from the domestic environment and the consumer s hands and body fluids. Microorganisms such as bacteria, yeasts and moulds are readily introduced when fingers are dipped into cosmetics 3. Microorganisms are adapted to a wide range of environments and can degrade a host of cosmetics product. The presence of pathogenic organisms and high levels of microorganisms in cosmetic products can cause physical deterioration of the products and can be a health hazard for consumers 4, 5. Infections of the eye and skin particularly of open wounds are the most typical public health hazards 6. The economic impact of such infections includes both the effect on the victim and the litigation against the manufacturer. Microbial contamination of cosmetics renders them unsuitable for use. While moldiness, colour change, frothing and packaging that bulges, leaks or explodes as a result of gas production are obvious effects of gross contamination, more subtle changes can occur. Active ingredients may also be rendered ineffective. Other contamination effects may be unpleasant aroma and viscosity changes which can occur which may also diminish the performance of the product. Body creams are cosmetic products for topical applications. They are esterified products of triglycerides and phospholipids from various vegetable oils, animals and natural fats 7. They are prone to contamination with Pseudomona aeruginosa, Burkholderia cepacia, Micrococci and Staphylococci including Staphylococcus aureus. Anerobic spore formers may also be found in these products. Fungal contaminants include Candida albicans and Penicillium 8. Though body creams contain preservative, microbiological spoilage still occurs. In some instances, micro-organisms in body creams, whether present initially or transfered to them, have been implicated as aetiological agents of diseases. The aim of this study therefore is to isolate, characterize and identify the fungal contaminants of some commercial body creams in Eke Awka Market, Awka, Nigeria. MATERIAL AND METHODS Samples Collection Five brands of Commercial body creams were purchased randomly from their dealers at Eke Awka Market in Awka Anambra State, Nigeria and transported to the microbiology laboratory of Nnamdi Azikiwe University, Awka, Nigeria for fungal isolation, characterization and identification. Fungal Isolation 380
Mycological Quality of Some Commercial Body A loopful of each of the samples was inoculated onto a petri dish containing sterile sabouraud dextrose agar by streaking. The petri dish was incubated at room temperature for seven days after which growth was observed and the colonies counted and recorded. Pure cultures of the isolates were obtained by repeated sub culturing on sterile sabouraud dextrose agar plates using sterile wire loops. The isolates were later stored on sterile sabouraud dextrose agar slants for characterization and identification tests. Characterization and Identification of the Isolates The isolates were identified on the basis of their cultural and microscopic features. Microscopic examination was carried out using lactophenol cotton blue solution. A drop of the solution was made on a clean, grease-free slide and a fragment of the test fungus was removed and introduced into the solution with the aid of a sterile needle. The isolate was thereafter spread properly on the slide and a cover slip placed on it gently, avoiding bubbles. The slide was then mounted and examined under the microscope. Identification was carried out following the description of Oyeleke and Manga 7 for the comparison of the fungal morphology. RESULTS The fungal counts of the body creams examined are shown in Table 1. Table 1. Fungal counts of the Body Creams Sample Number of colonies 1 20 2 12 3 10 4 22 5 18 They ranged between 10 and 22 colony forming units per gram. Sample 4 had the highest fungal colonies (22) while sample 3 had the least (10). The cultural and microscopic characteristics of the fungal isolates are presented in Table 2. Isolate Table 2. Cultural and Microscopic Characteristics of the Isolates Cultural Microscopic Characteristics Characteristics 1 Colonies were green and velvety 2 Colonies were pink and cottony Conidia were green and globose. Conidiophores were simple, long and erect. Hyphae were septate and branched. Spore heads were brush like with long chains of conidia Microconidia and macroconidia were seen. Microconidia were ovoid. Macroconidia were borne on phialides Identity Penicillium sp Fusarium sp 3 Colonies were grey Sporangia were black with zygospores. Mucor sp
Onuorah et al., 2014 4 Colonies were white and cottony 5 Colonies were black with white edges Sporangiospores were long and branched. Hyphae were non-septate and branched Sporangia were ellipsoidal Conidia heads were large, globose, darkbrown and biseriate. Conidia were globose and rough-walled. Conidiophores were smooth-walled. Rhizopus sp Aspergillus sp They were identified as Penicillium sp, Fusarium sp, Mucor sp, Rhizopus sp and Aspergillus sp. The occurrence of the fungi in the body creams is shown in Table 3. Table 3. Occurrence of the Isolates in the Body Creams Sample Penicillium sp Fusarium sp Mucor sp Rhizopus sp Aspergillus sp 1 + - + - + 2 - - + + + 3 - - + - - 4 - + - - + 5 + - - - - Aspergillus sp and Mucor sp occurred in three of the samples studied while fusarium sp and Rhizopus sp had the least frequency of occurrence. The study showed that the body creams sampled were contaminated by fungi. Most of the isolated fungi had been reported in body creams 8, 9. Nasser 9 reported Aspergillus as the most common genus found in body creams. Brooks and Antai 3 also made a similar observation. Hugbo et al., 10 studied the microbial quality of body creams and isolated Aspergillus fumigatus and Penicillium sp. Michael et al., 11 reported that fungi are the major contaminants of body creams. Jasnow and Smith 12 reported that fungal contaminants in unused cosmetic body creams are due to the quality of the raw materials used in its manufacture. The isolation of these fungi from the body creams poses some health concern particularly Aspergillus sp which had been isolated from mycotic abcesses of humans by Sukroongreung and Thakerngpol 13. Cosmetic ingredients supply nutrients for microbial growth, therefore body creams should be produced in a hygienic environment. Product premises, equipments, instruments, storage tanks and containers should be maintained in a high standard of cleanliness and guidelines of good manufacturing practices should be observed by manufacturers. All starting materials should correspond to agreed standards and be of good quality. DISCUSSION The presence of fungi in the body creams studied showed that there was contamination in one or more of the raw materials used in their manufacture. Good manufacturing practices must always be followed and raw materials, 382
Mycological Quality of Some Commercial Body particularly those of natural origin must be tested for contamination before use and limit of acceptability established. REFERENCES 1. Brian, P. (2011). Cosmetics Microbiology. Microbiology Today, 28: 185-187. 2. Orth, D.S. (1989). Handbook of Cosmetics. The Microbiology of Cosmetics. Marcel Dekker. 147-158. 3. Brooks, A.A. & Antai, S.P. (2006). Bacteriological Status of some Cosmetic products in three states of Southern Nigeria. Nigerian Journal of Microbiology, 20(3): 1345-1354. 4. Behravan, J., Bazzaz, F. & Malaekeh, P. (2005). Survey of bacteriological contamination of Cosmetic Creams in Iran. International Journal of Dermatology, 44: 482-485. 5. Campana, R., Scessa, C., Patrone, V., Vittoria, E. & Baffone, W. (2006). Microbiological Study of Cosmetics products during their use by consumers: health risk and efficacy of preservative systems. Letters of Applied Microbiology, 43: 301-306. 6. Klotz, S.A., Christopher, C., Penn, J., Gerald, J.N. & Salim, L.B. (2000). Fungal and Parasitic infections of the eye. Clinical Microbiology Reviews, 4:662-685. 7. Oyeleke, S.B. & Manga, S.B. (2008). Essentials of Laboratory Practice in Microbiology. Tobest Publishers, Minna, Nigeria. 36-75. 8. Madden, J.M. & Jackson, G.J. (1981). Cosmetics preservation and microbes: view point of the food and drug administration, cosmetics and toiletries, 96: 75-77. 9. Nasser, L. (2008). Fungal profiles isolated from open and used cosmetic products collected from different localities in Saudi Arabia. Saudi Journal of Biological Sciences, 15 (1): 121-128. 10. Hugbo, G.P., Onyekwelu, O.A. & Igwe, I. (2003). Microbial contamination and preservation of some brands of cosmetic creams. Tropical Journal of Pharmaceutical Research, 2: 229-234. 11. Michael, M.D., Juliet, N.O. & Jospehine, M.A (2011). Microbiological quality assessment of some brands of cosmetics powders sold within Jos metropolis, Plateau State. Journal of Microbiology and Biotechnology Research, 1(2): 101-106. 12. Jasnow, S.B. & Smith, J.L. (1975). Microwave Sanitization of color additives used in cosmetics feasibility. Applied Microbiology, 2: 205-211. 13. Sukroongreung, S. & Thakerngpol, K. (1985). Abnormal form of Aspergillus terrus isolated from mycotic abcesses. Mycopathologia, 91: 49-52. Madden, J.M and Jackson, G.J. (1981). Cosmetics preservation and microbes: view point of the food and drug administration, cosmetics and toiletries, 96: 75-77.