Evaluation of Locally Sourced Raw Materials for Use As a Microbiological Media

J. Biol. Today's World. 2013 Jan; 2 (1): 1-4 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ Journal of Biology and Today's World ISSN 2322-3308 http://www.journalbio.com Received: 02 November 2012 • Accepted: 29 November 2012 Research doi:10.15412/J.JBTW. 01020101 Evaluation of Locally Sourced Raw Materials for use as A Microbiological Media Abalaka M.E1*, Daniyan S.Y1, Adeyemo S.O2, Okolo M.O3, Abdulsalam R1 1 Department of Microbiology, Federal University of Technology, Minna, Nigeria 2 Department of Biochemistry, Bingham University, Karu, Nigeria 3 Department of Microbiology, Kogi State University, Anyigba, Nigeria *correspondence should be addressed to Abalaka, Department of Microbiology, Federal University of Technology, Minna, Nigeria; Tell: +2348036503689; Fax: +23; Email: [email protected]. ABSTRACT Local prepared nutrient media made from guinea corn and maize were used to culture bacteria such Staphylococcus aureus, Klebsiellasp, Salmonella typhi, Escherichia coli, and Pseudomonas aeruginosa and compared with the growth on commercial nutrient agar and broth. Bacteria growth was higher in the commercial nutrient agar and broth but with little significant difference. Locally prepared potato dextrose agar was also used to culture fungal isolates such as Aspergillusniger, Aspergillusfumigatus, Aspergillusflavus, Cephalosporiumsp, and Mucorsp and compared with commercial made potato dextrose agar using mycelia extension technique. Fungal growth was higher in commercially prepared potato dextrose agar with little difference. The results of these studies suggest that these locally sourced materials could be used for media preparation in place of commercially produced media. Key words: Microbiological Media, Locally prepared Nutrient Media, Bacteria, Fungi Copyright © 2014 Abalaka M.E et al. This is an open access article distributed under the Creative Commons Attribution License. 1. INTRODUCTION precisely tailored to select organisms with very properties medium is solid or a liquid preparation containing (3). Such media are very useful as a simple complex materials for the culture (growth) of medium may be sufficiently rich to completely meet the Amicroorganisms, animal cells or plant tissue nutritional requirement of many different microorganisms. cultures (1). To culture organisms in laboratory, it requires In addition, complex media often are needed because the the preparation of substances which can be used as food. nutritional requirements of a particular microorganism are Such nutrient preparations which microorganisms can use unknown and thus, a defined medium cannot be as food are called culture media. Different microorganisms constructed. This is the situation with many fastidious require different nutrient materials. Thus, culture media bacteria that have complex nutritional or cultural vary in form and in composition depending on the species requirement; they may even require a medium containing to be cultivated (2). A medium may be formulated as either blood serum (4). Complex media contain undefined permissive with the intent of allowing the growth of component like peptone, meat extract and yeast extract. whatever organisms are present, or restricted or selective Peptones are protein hydrolyzate prepared by partial with the intent of only selecting for growth only a proteolytic digestion of meat, casein, soy malt, gelatin and particular subset of those organism (2). This may take the other protein sources. They serve as source of carbon, form of a nutritional requirement, for instance providing a energy and nitrogen. In composition, peptones are mixtures particular component such as lactose as the only source of containing only partly known concentration and identify, a carbon for energy including a particular antibiotics or other variety of peptides and polypeptides, protease, amino acids substances in order to select only organisms which are and carbohydrate including inorganic and many organic resistant to that substances. This correlates to some degree micronutrients. Peptone provides insoluble and assimilable with defined and undefined media are made from natural form of all the phosphorous and sulfur and essential product and containing an unknown combination of very mineral content of living materials as well as the organic many organic molecules, while defined media can be carbon and nitrogen source [4]. Beef extract contains 1 J. Biol. Today's World. 2013 Jan; 2 (1): 1-4 ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ amino acid, peptides, nucleotides, organic acids, vitamins plates were incubated at room temperature, and the growth and minerals. Among meat extract and infusions beef was examined using radial extension method at 48 and 72 extract and aqueous meat infusion made by infusing hours. (soaking) fresh ground meat in water are common ingredient of media useful for many species. Beef 3. RESULTS AND DISCUSSION infusions are rich in mineral, organic micronutrient, 3.1. Bacterial count on locally prepared and commercially proteins, protein derivatives and carbohydrates. They are prepared nutrient agar often supplemented with 1% peptone. Culture media fluids Table 1 show that the counts of some selected bacteria i.e made from beef extract as extract broth. All such Staphylococcusaureus, Klebsiellasp, Escharichia coli, nutritional media including simple solutions of peptone are Salmonella typhi and Pseudomonasaeruginosa on locally sometimes loosely included in the general terms nutrient prepared nutrient agar using guinea corn and meat extract. broth (5). Escherichia coli had the highest microbial count on the guinea corn agar with a mean growth of 7.1 + 0.1 when 2. MATERIALS AND METHODS compared with the maize agar, followed by Klebsiellasp 2.1. Collection of Samples with a mean growth of 4.2 + 0.3 on guinea corn agar. Maize, guinea corn and Irish potato was obtained from a Staphylococcus aureus had the lowest count with the mean retail outlet in Bosso area of Minna, Nigeria and was growth of 2.05 + 0.01. Commercial prepared nutrient agar transported to the Microbiology laboratory of federal had the highest microbial count when compared with the university of Technology, Minna, Nigeria. Meat (beef) was guinea corn and maize agar. obtained from Minna Abattoir along western bypass and was equally transported to the laboratory. Table 1. Count (cfu/ml) of some selected bacteria on locally prepared nutrient agar using guinea corn and meat extract Organism GCA (x105cfu/ml) MA (x105cfu/ml) CPN (x105cfu/ml) 2.2. Processing of Materials Two hundred grams of maize and guinea corn was washed Staphylococcus 1.005 + 0.01 1.05 + 0.05 2.15 + 0.05 with distilled water and manually grinded using a clean aureus grinder. It was then sieved using sterile filter cloth into Klebsiellasp 4.20 + 0.30 4.15 + 0.15 7.20 + 0.10 Escherichia coli 7.10 + 0.10 4.95 + 0.05 2.30 + 0.16 500ml beaker and covered with aluminum foil. The meat Salmonella typhi 3.55 + 0.55 2.60 + 0.10 1.35 + 0.05 (beef) was boiled for 30 minutes and the extract was Pseudomonas 1.06 + 0.04 1.10 + 0.10 1.05 + 0.05 transferred into a sterile beaker and covered with aeruginosa aluminum foil. Two hundred ml of guinea corn and maize Key:GCA –Guinea corn agar; MA – Maize agar; CPNA – Commercial prepared nutrient agar extract each was mixed with 100ml of meat extract, 20g of agar-agar, 5g of sodium chloride and the medium was 3.2. Bacterial count on locally prepared and commercially sterilized at 121oC for 15 minutes.Two hundred grams of prepared nutrient broth Irish potato was boiled with water for 30 minutes, and the Table 2 shows the count of some selected bacteria extract is allowed to cool and transferred to a 500ml beaker Staphylococcus aureus, Klebsiellasp, Escharichia coli, and was covered with aluminum foil. 20g of agar-agar, 10g Salmonella typhi and Pseudomonasaeruginosa on locally of glucose was dissolved in a 500ml beaker containing prepared nutrient broth using maize extract and meat Irish potato extract and 0.2g of chloramphenicol powder extract Pseudomonas aeruginosa had the highest microbial was added to inhibit the growth of bacteria. Each of the count on the guinea corn broth with a mean growth of 8.10 mixture was sterilized using a validated autoclave at a + 0.10 when compared with the maize broth followed by temperature of 121oC for 15 minute and it was dispensed Klebsiellasp with a mean growth of 5.05 + 0.05 on guinea into petri-dish and labeled accordingly. corn broth. Escharichiacoli had the lowest count with the mean growth of 3.10 + 0.10. Commercially prepared 2.3. Inoculation of Organisms nutrient broth had the highest microbial count when Pure isolates of each of Staphylococcus aureus, compared with the guinea corn and Mize agar. Klebsiellasp, Salmonellatyphi, Escherichia coli and Table 2. Count (cfu/ml) of some selected bacteria on locally prepared nutrient broth using Pseudomonas aeruginosa was sub-cultured into guinea corn and maize extract commercially prepared nutrient media and locally prepared Organism GCA (x105cfu/ml) MA (x105cfu/ml) CPN(x105cfu/ml) Staphylococcus 4.05 + 0.05 4.10 + 0.10 2.15 + 0.05 medium using a pour plate technique. The plate was aureus o incubated at 37 C for 24 hours and the growth rate in the Klebsiellasp 5.05 + 0.05 1.95 + 0.05 1.15 + 0.05 commercial nutrient

Load more