www.ijird.com September, 2016 Vol 5 Issue 10 ISSN 2278 – 0211 (Online) Microorganisms Associated with Composting of Pycnanthus angolensis with Cow Dung Olabode, O. O. Senior Lecturer, Integrated Science Department, Adeyemi College of Education, Ondo, Nigeria Dr. Adegunloye, D. V. Senior Lecturer, Department of Microbiology, Federal University of Technology, Akure, Nigeria Akinyele, B. J. Professor, Department of Microbiology, Federal University of Technology, Akure, Nigeria Akinyosoye, F. A. Professor, Department of Microbiology, Federal University of Technology, Akure, Nigeria Abstract: Composting and vermicomposting processes were carried out using wood dust of Pycnanthus angolensis with animal waste added as boosters for compost. Substrates were prepared with varying ratio of wood dust: animal waste and kept for a period of 10 weeks on shelf outdoor. The temperature reached a maximum of 42.5 0C in composting in 2 weeks of the process and dropped to 25.5 0C in vermicomposting at the same period. At the initial stage, more of gram positive bacteria were isolated, including Bacillus coagulans, Pediococcus cerevisae, Streptococcus faecalis. Lactobacillus subger, Micrococcus luteus, Actinomyces bovis, Actinomyces ericksonii. Some of the gram negative bacteria isolated were Acinetobacter anitratis, Citrobacter freundii, Escherichia coli Pseudomonas aeruginosa and Brucella militenses. The fungi species isolated include Alternaria alternata, Aspergillus flavus, Aspergillus fumigatus, Cladosporium sp Curvularia The highest total population of bacteria was 2.55×10 7 cfu/ml, while that of fungi was 4.35 10 5 sfu/ml isolated from compost substrate of the wood dust and cow dung. The total bacteria population increased from 4.55 ×10 6 to 2.55×10 7cfu/ml in composting, and 6.82×10 6cfu/ml in vermicomposting in 2 weeks. Both bacteria and fungi diversity and population reduced significantly at the end of the process with the most occurring genus as Bacillus and Aspergillus species Keywords: Animal waste, bacteria, decomposition, isolation, substrate 1. Introduction Composting is a natural process of recycling organic material, by the action of biodegradative microorganisms into nutrient rich product known as compost. The organic waste materials mainly of animal and plant origin are potential sources of organic matter and plant nutrient (Adeniran et al ., 2003). Animal wastes in particular are highly nitrogenous and rich in mineral elements, such as potassium, calcium and phosphorus. They are used as soil enrichment in plant cultivation. Composting is a natural biological process resulting in the controlled decay of organic matter in a warm moist environment by bacteria, fungi, and other organisms (Devi, 2012). The organic waste materials mainly of animal and plant origin are potential sources of organic matter and plant nutrient (Adeniran et al ., 2003). Animal wastes in particular are highly nitrogenous and rich in mineral elements, such as potassium, calcium and phosphorus. They are used to enrich the soil in plant cultivation. Composting is a process of solid-waste fermentation which explores the biodegradative ability of microorganisms and mineralisation. It is of importance to a mushroom grower because it affords the preparation of substrates that promote the growth of mushroom but remove pathogenic microorganisms, reduce infestation by insects, fungi, and thereby improve the yield of mushroom fruit body (Gbolagade, 2006). For a long time, composting is applied as a biological process of organic waste management in many parts of the world, but recently the use of certain species of earthworm have become of tremendous importance and advantage in vermicomposting for biological degradation of organic wastes to obtain fertilizer. The biochemical decomposition of organic matter is primarily accomplished by microorganisms, but earthworms are crucial drivers of the process as they may affect microbial decomposer activity by grazing directly on microorganisms (Aira et al., 2009; Monroy et al., 2009; Gomez-Brandon et al., 2011). INTERNATIONAL JOURNAL OF INNOVATIVE RESEARCH & DEVELOPMENT Page 176 www.ijird.com September, 2016 Vol 5 Issue 10 Microorganisms which colonize mushroom compost during composting process are the crucial determinant of the nutrient status of compost that is required for mushroom cultivation. For the composting process, the importance of microbial communities is well established (Ryckeboer et al. , 2003). Previous studies have established the importance of microbial communities in the composting process. These have included bacterial population, actinobacteria and fungi (Chandna et al., 2013). Bacteria and fungi in particular, digest the complex organic compound that make up living matter and reduce them to simpler compounds that plants can use for food (Oyeyiola et al ., 2013). The biomass ratio of fungi to prokaryotes in compost is about 2:1. In addition, fungi existing in compost use many carbon sources, mainly lignocellulosic polymers and can survive in extreme conditions. They mainly are responsible for compost maturation. Fungal genera (e.g. Aspergillus, Penicillium , Fusarium, Acremonlum and Cladosporium ) are dominant in the compost process (Anastasi et al ., 2005). There are parameters that play an important role in the frequency of fungal species during a composting process including temperature, moisture, C/N ratio, pH and required oxygen to maintain the aerobic conditions. Two parameters of moisture and pH are more effective in fungal activities (Grigatti et al., 2011). Gbolagade (2006) identified eleven bacteria isolates from substrate of Terminala superba wood dust supplemented with wheat bran. The types of substrate, environmental condition would determine the type of bacteria that can colonize it. The backbone of organic agriculture is the maintenance of good soil and healthy environment through the use of compost. Cultivation of saprophytic edible mushroom may be the only currently economical biotechnology for lignocellulose organic waste recycling that combines the production of protein rich food with the reduction of environmental pollution (Obodai and Apetogbor, 2003). Mushroom is cultivated worldwide because it is able to utilise lignocellulose and grow on various agricultural wastes (Mshandete and Cuff, 2008 .). Various wastes from agricultural industry could be used to prepare compost for mushroom cultivation (Gbolagade et al ., 2011). Lignocellulosic wastes from tree wood dust are abundantly available in Nigeria for composting. Composting organic matter wastes is an important pathway for carbon flow and cycling of nutrients, both in industrial and developing countries (Bonito et al ., 2010). Bacteria associated with varying compost and vermicompost preparation using animal waste as booster has not been determined. The objective of the present study is to provide more information on the microorganisms’ types and load responsible for the composting of Pycnanthus angolensis with cow dung, for compost preparation that could be used for mushroom cultivation. 2. Materials and Methods 2.1. Collections of Animal and Agricultural Wastes Cow dung was collected inside sterile polythene bags from animal farm in Pele village located in Ondo West Local Government Area of Ondo State, Nigeria. Sawdust of Pycnanthus angolensis (Africana nut) was collected in clean bags from sawmills at Oka, Ondo where the wood was milled into 5mm sizes. Wastes were transferred to the laboratory for composting and vermicomposting preparation. 2.2. Compost Preparation The composting was prepared by passive pile method (Keith et al ., 2009). One kilogram of composting substrate was prepared by mixing wood waste of Pycnanthus angolensis and cow dung at ratios of 5:5,6:4,7:3,8:2,9:1 (numbered as 1,2,3,4 and 5) for substrates, the control sample as 100% wood dust. The various samples were composted in plastic bowls of 30cm depth and observations were made daily for 70days. The composting was kept at moisture of 65% and ambient room temperature. Temperature of the core were taken daily during the period of the experiment. Samples of the composting were taken daily for bacteriological analyses. 2.3. Vermiculture Eudrillus eugiene identified as a fast breeder and active feeder on organic matter that are high in nitrogen was used for the vermiculture (Jambhekar 1992). Earthworm culturing was done under shelter to avoid direct sunlight and heavy downpour using fifteen litre plastic buckets with perforated lid. A bed of 10cm height using sawdust as the base was sprinkled with water to get a moisture level of 40-45% which made the bed to appear wet. Different substrate preparation of Pycnanthus angolensis were mixed with the cow dung in equal quantity with appropriate quantity of water to make a homogenous mixture. The mixture was kept for two weeks, while the material was turned 2 to 3 times at 4-5 days interval. This was transferred on the layer of beddings prepared earlier. Worm was introduced into the prepared culture. The worm fed actively on the organic matter and bred (Henamgee, 2003). 2.4. Vermicomposting Compostings heaps of the various substrates were made inside plastic bowl at the rate of 10 worms/kg of feed mix (substrate). It was kept wet to a moisture level of 70% for 60 days (Henamgee, 2003). The vermicomposting formed completely gave the smell of moist soil. Samples were taken during the period of vermicomposting for 70 days for bacteria isolation and identification. Colonial characteristic of the bacterial isolates were