Isolation of Ascomycetous Fungi from a Tertiary Institution Campus Soil
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JASEM ISSN 1119-8362 Full-text Available Online at J. Appl. Sci. Environ. Manage. December, 2008 All rights reserved www.bioline.org.br/ja Vol. 12(4) 57 - 61 Isolation of Ascomycetous Fungi from a Tertiary Institution Campus Soil DUROWADE, K. A1; *KOLAWOLE, O. M2; UDDIN II, R. O3; ENONBUN, K.I2 1Department of Epidemiology and Community Medicine, College of Medicine, University of Ilorin. Kwara State. 2Department of Microbiology, Faculty of Science. University of Ilorin. P.M.B 1515, Ilorin, Kwara State. Tel: 08060088495. Email: [email protected] 3 Department of Crop Protection, Faculty of Agriculture, University of Ilorin. Kwara State ABSTRACT: Studies were carried out on the ascomycetous fungi present in six different but carefully selected sites on the University of Ilorin permanent site soil. Fungi isolation was done by the soil dilution method incubated at 27oC for 72 hours. The predominant Ascomycetous fungi isolated include among others; Aspergillus niger, Fusarium solani, Fusarium oxysporum, Penicillium italicum, Fusarium acuminatum, Fusarium culmorum, Candida albicans, Botrytis cinerea, Geotrichum candidum, Trichoderma viride, Verticillium lateritum, Curvularia palescens, Penicillium griseofulvum, Penicillium janthinellum, Penicillium chrysogenum, Aspergillus terreus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus glaucus, Aspergillus clavatus, Cladosporium resinae, Alternaria alternate, Trichothecium roseum, Phialophora fastigiata, Aspergillus nidulans, Aspergillus wentii, Humicola grisea, Trichophyton rubrum, Helminthosporium cynodontis, Penicillium funiculosum, Penicillium purpurogenum, Saccharomyces cerevisiae, Trichoderma harzianum, Scopulariopsis candida.The physico- chemical characteristics of soil samples was found to affect the distribution and population of fungi. The colony count in the study are ranged between 5.8 x 10″ per gram of soil to 1.63 x 10″ per gram of soil. The soil consists of high organic matter content. @ JASEM Soil contains a vast array of microorganisms such source of Carbon, Nitrogen, Vitamins and trace as bacteria, viruses, fungi, actinomycetes, protozoa elements also determine the rate of spore and algae (Alexander, 1977; Olowonihi, 2003). development under natural (soil) conditions Soil organism participates in the genesis of the (Michael and Donald, 1996; Ivan et al., 2008). It habitat, which they live. They, together with the has been discovered that micro-organisms play an total biota and especially the higher vegetation, important role in the fertility of the soil, their constitute one of the five interactive factors in soil abundance and existence is solely hung on the formation; the other four are climate, topography, environmental (soil) conditions (Paul and Clerk, parent material, and time (Beare, 1997). The 1989). This study intends to examine the physico- physical and chemical breakdown of rocks to fine chemical characteristic of the soil samples and to particles with large surface areas and the identify as many Ascomycetors fungi as possible accompanying release of plant materials initiate the present in the six different sites of soil collected soil forming process (Paul and Clerk, 1996). Two from the University of Ilorin permanent site. major nutrients that are deficient in the early stages of the process are carbon and nitrogen therefore, MATERIALS AND METHODS the initial colonizer of soil parent material are often Study Area: This study was carried out in the organisms capable of enhancing photosynthesis and permanent site of the University of Ilorin (8029N, nitrogen fixation. These are predominantly the 4035E) .Ilorin is on southern guinea savanna cyanobacteria, also known as the blue green algae ecological zone in Nigeria. Between the months of (Sylvia et al., 1997). After higher vegetation has February to November rainfall distribution pattern become established, a continuum of soil processes of the land is bimodal; with the first major peak produces the dynamic mixture of living and dead between May and mid-August while the second cells, soil organic matter (SOM), and mineral peak is between September and the mid-November. particles in sufficiently small sizes to permit the The average rainfall for Ilorin is 1000-1240 intimate colloidal interaction characteristic of soil Selection of sample sites: Suitable sites were (Lambart et al., 1979; Andrew et al., 2008). The selected within the study area. Six sites were Ascomycetes (Sub-division Ascomycotina) chosen they includes: constitute the largest class of fungi, the number of Site 1; Academic Area: known species being approximately 32,000. There This is the area around the lecture halls, is considerable diversity of form and structure administrative buildings etc.There is constant among the Ascomycetes. At one end of the scale movement of students and academic staff in this are the truffles and morels (Adamas et ai., 1999). area. It is often littered with paper wastes from The Ascomycetes derive their name from the ascus, food eaten by students. the sac-like vessel which contains the sexual spores Site 2: Clinic: (ascospores).The ascus is a walled receptacle This is the area around the school Health centre enclosing the spores and usually rupturing at the where staff and students get their medical attention. physical and nutritional factors where a Site 3: Primary School (University of Ilorin Staff pronounced affect on sporulation of fungi School): occur.Temperature, light, hydrogen ion This is the study environment of the primary school concentration, aeration and humidity are some of pupils. It is often characterized with the movement the important physical factors. Factors such as * Corresponding author: Durowade, K. A1 Isolation of Ascomycetous Fungi from a Tertiary Institution Campus Soil 58 of children and it is often littered with paper and Plates were then incubated for 72 hours at room food remains of the children. temperature (27oc) for fungal growth. Site 4: Senior Staff housing: Preparation of Stock Culture of Fungal Isolates: This is the residential area of the senior staff of the The pure cultures of the different isolates obtained university. The area is characterized with children were sub-cultured into sterile McCartney bottles playing in their courtyard. It is often littered with containing agar slant. These bottles were also domestic waste, used papers, dead leaves and incubated at 27oC until appreciable mycelia growth grasses. and fungal sporulation were observed.These were Site 5: Reservoir: kept as stock cultures. This is the area around the water storage system in Characterization and Identification of the University. There is a constant movement of Ascomycetous fungal Isolates: Colonial water tankers and staff in the area. It is often morphology and microscopic examinations of the littered with dead leaves and grasses and food various isolates of pure cultures were used to remains of the people coming into the area. determine the reproductive and vegetative Site 6: Student housing: structures. Consequently, identification was done This area covers the halls of residents of the using Onions et al., (1981). Spore identification University students. It is characterized with the to was achieved by reference to Spore atlases of and fro movement of the students in the halls. It is Gregory (1973) and Anna (1990). often littered with used papers, dead leaves from trees and domestic wastes. RESULTS AND DISCUSSION Collection of soil samples: Soil samples were The results based on the physico-chemical collected employing sterile soil augers, hand trowel characteristics of the soil samples viz: water and polythene bags (Akinyanju and Fadayomi, holding capacities, moisture contents, organic 1989). The soil was dug out using augers up to 0- matter contents, and pH values is shown in Table 1. 20cm depth and was immediately scooped into The results revealed that site 3 had the highest sterile polythene bags using the hand trowel. The water holding capacity of 0.43ml/g, while site 2 samples were collected from 2 spots in each site had the lowest with 0.3ml/g. The range value was and then mixed together in order to obtain a 0.13ml/g. This finding could possibly be correlated representative sample. to the sandy content in the soil sample from site 2 and the clay content in the soil sample in site 3. PHYSICO-CHEMICAL ANALYSIS Site 4 had the highest moisture content of 1.56%, Soil Water Holding Capacity Determination: while the least value of moisture content obtained Soil sample which has been dried in the oven at goes to site 2 with 0.12%. In the case of organic o 105 C for 24 hours was used for the water holding matter contents, site 6 had the highest with 4.76% capacity determination according to the method of and the lowest value goes to site 5 with 2.63%. Pramer and Schmidt (1964). Based on the pH values of the sampled soil, it was Soil Moisture Content Determination: The found that site 4 was acidic with a pH value of 5.7 method used for this determination was that of while site 1 recorded a high pH value of 7.4 Pramer and Schmidt (1964). indicating that the soil was alkaline. This range of % organic matter (humus) in sample = pH values affected the type of fungi found in the weight of humus x 100 different sites. The various ascomycetous fungi Weight of dry soil taken isolated in all the six sites were identified and are represented in Table 2. In all, thirty five (35) Soil pH Determination: About 20g of air-dried ascomycetous fungi were isolated.The results based sieved soil into a 100ml beaker and 20ml of sterile on the different colony counts of each dilution distilled water was added. The suspension was left values (i.e. 10-1, 10-2, 10-3 and 10-4) of each site is to stand for 30 minutes with occasional stirring to shown in Fig 1. The average number of colonies of enhance equilibrium reaction. The pH of the each site based on the varying dilution was suspension was taken by inserting the glass calculated and results showed that site 3 had the + hydrogen (H ) electrode of the pH meter (Pye highest number of 5.8 x 10″ per gram of soil while 2 Unicam, model 292mk pH meter) into the partly site 5 recorded the lowest number of 1.63 x 10″ per settled suspension.