International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 3(11), 28-31, November (2014) Int. Res. J. Biological Sci. International Science Congress Association 28 Study of Prevailing of Deuteromycetous Fungi on the Petro-Polluted Soil Benal T., Shivani K., Pagare R.L. and Chitnis S. Department of Botany Govt. Holkar Science College Indore 452001, INDIA Available online at: www.isca.in, www.isca.me Received 26th June 2014, revised 7th September 2014, accepted 10th October 2014Abstract The present paper investigates deuteromycetous fungi on the petro-polluted soil. These are conducted by the enumeration of the fungal population and the identification. Fungus observation physico-chemical analysis of soil and identification of the soil fungi. The soil sample examined for isolating fungi from petroleum contaminated site. Identified as Aspergillus, Penicillium, Fusarium, Rhizopus, Mucor, Cladosporium, Morssonina Chaetomium, Curvularia, Helminthosporium, Alternaria and Trichoderma Species. The isolation of filamentous fungi in environments containing oil or its sub products found a very similar diversity of genera to that found in our study. Petro-polluted site the growth of fungal diversity was higher due to more carbon concentration. Keywords: Soil, Petro-pollution, aspergillus, penicillium, fusarium, rhizopus. Introduction Petroleum hydrocarbons of soil contamination are one of the world’s most common environmental problems. Petroleum hydrocarbons (TPHs) are one of the most common groups of persistent organic contaminants. Crude petroleum and environmental conditions crude petroleum oil from petroleum contaminated soil from North East India. With the development of the economy and petroleum exploration, the contamination of soil with petroleum compounds is a concern worldwide. Petroleum like all fossil fuels primarily consists of a complex mixture of molecules called hydrocarbons. Carbon is so popular among various elements that is has a separate branch of chemistry called organic chemistry it is common element of organic world. The hydrocarbon molecules that make up crude oil and petroleum products are highly toxic to many organisms, including humans. The objective of this investigation is to assess the impact of various concentrations on the fungal population of a soil in the Niger Delta prone to crude oil pollution. This is conducted by the enumeration of the fungal population (heterotrophic and hydrocarbon-degrading fungi) and the identification of both heterotrophic and hydrocarbon-degrading fungi (types or diversity) in control and in crude oil polluted soils. The present paper deals with prevailing of deuteromycetous fungi on the petro-polluted soil. Material and Methods Soil dilution method: 1gm of soil sample was suspended in 10ml of double distilled water to make soil suspensions (10-1 to 10-5). Dilution of 10-3, 10-4 and 10-5 were used to isolate fungi. 1 ml of soil suspension of each concentration were added to sterile Petri dishes (triplicate of each dilution) containing 15 ml of sterile Potato Dextrose Agar and Czapek,s Dox Agar. One percent streptomycin solution was added to the medium before pouring into petriplates for preventing bacterial growth. The Petri dishes were then incubated at 28 ± 20 C in dark. The plates were observed everyday up to three days. Physico-chemical analysis of soil: The collected soil was characterized for its physico-chemical properties. The physico-chemical parameters were measured by standard methods. Physical and chemical parameters of soil such as pH, salinity, organic carbon, nitrogen, phosphorous and potassium were analyzed. The physic chemical parameters of the soil samples were analyzed at Soil Testing Laboratory (STL), From Agriculture College of Indore (MP). Methods of microscopic examination: For light microscope the optical equipment used were dissecting microscope, research microscope (10x and 15x eye pies and 10 x to 100x objectives), equipment for microphotography, camera Lucida and ocular and stage micrometers. Identification of the soil fungi: Fungal morphology were studied macroscopically by observing colony features (Colour and Texture) and microscopically by staining with lacto phenol cotton blue and observe under compound microscope for the conidia, conidiophores and arrangement of spores .The fungi were identified with the help of literature. Soil Sampling and Analysis: The most common and economical method for sampling an area is composite sampling, where sub-samples are collected from randomly selected locations in a field, and the sub-samples are composite sampling provide average values for the sampled area. The actual number of sub-samples depends on field size and uniformity. A soil sample analysis has done soil testing laboratory in the Agriculture college of Indore (MP). International Research Journal of Biological Sciences ________________________________________________ Vol. 3(11), 28-31, November (2014) International Science Congress Association Results and Discussion The soil samples examined for isolating fungi from petroleum hydrocarbon contaminated site in Manglia depo dilution technique were identified as Aspergillus Fusarium, Rhizopus, Mucor, Cladosporium, Morssonina Chaetomium, Curvularia, Helminthosporium, Alternaria Trichoderma species. Studies on the isolation of filamentous f ungi in environments containing oil or its sub products found a very similar diversity of genera to that found in our study, such as: Aspergillus and Penicillium4, 5. Recently, it was recorded that the genera of fungi such as Penicillium, Aspergillus, Fusarium, Rhizopus associated with petroleum hydrocarbon contaminated soil. In their studies they isolated Penicillium and Aspergillus From hydrocarbon contaminated soil and identified as hydrocarbon degrading fungi along with Tricoderma, Rhizopus sp. The similar results of our study were also obtained by Obire et al. , in their studies on effect of different concentrations of crude oil on fungal populations of soil. They isolated fourteen fungal genera from soil. These include Alternaria, Aspergillus, Candida , Cladosporium, Fusarium, Geotrichum Mucor Rhizopus, Rhodotolura, Saccharomyces , Trichoderma . The fungal isolates obtained in their study were mainly Aspergillus species, while ot hers were Penicillum, Rhizopus and Rhodotorula species which were all able to utilize hydrocarbon as carbon source 7 filamentous fungi from soil, which were able to degrade crude oil. Fusarium and Aspergillus sp . They isolat hydrocarbon degrading fungi from hydrocarbon contaminated soil. Aspergillus species were isolated form soil polluted by petroleum products in cross River University of Technology, Calabar, Nigeria. Comparison between depot soil and gram field soil (pH, EC&Carbon)  \n \r International Research Journal of Biological Sciences ________________________________________________ International Science Congress Association The soil samples examined for isolating fungi from petroleum hydrocarbon contaminated site in Manglia depo t Indore M.P. by Aspergillus Penicillium, Mucor, Cladosporium, Morssonina Helminthosporium, Alternaria and species. Studies on the isolation of filamentous ungi in environments containing oil or its sub products found a very similar diversity of genera to that found in our study, such Recently, it was recorded that the genera of fungi such as Rhizopus and Mucor are associated with petroleum hydrocarbon contaminated soil. In Aspergillus sp From hydrocarbon contaminated soil and identified as Tricoderma, Fusarium, sp. The similar results of our study were also obtained , in their studies on effect of different concentrations of crude oil on fungal populations of soil. They isolated fourteen fungal genera from soil. These include Cephalosporium, Geotrichum Mucor , Penicillium, , Torulopsis and . The fungal isolates obtained in their study were hers were Trichoderma, species which were all 7 . Isolated several filamentous fungi from soil, which were able to degrade crude . They isolat ed and identified hydrocarbon degrading fungi from hydrocarbon contaminated species were isolated form soil polluted by petroleum products in cross River University of Technology, In our present work, Aspergillus and present in dominant numbers. Our finding coincides with the work of Elisane et al.10, who also isolated four strains from the contaminated soil. They were identified as Aspergillus are the most commonly encounte hydrocarbon degraders in oil contaminated tropical soils, which are in agreement with the present work fungi from total hydrocarbon contaminated soil and identified by microscopy as Penicillium, A spergillus The different result from our findings were obtained isolated many fungal species that were able to degrade poly cyclic aromatic hydrocarbons. The species isolated were Coniothyrium fuckelii, Gliocadium virens Phialoph ora hoffmannii, Scopulariopsis brumptii Trichoderma harzianum along with genera were similar to our finding. With the serial dilution plate technique, Aspergillus and Penicillium species genera in the term of number. The growth rate of each fungus shows that Rhizopus sp. had the highest growth diameter in low petro contaminated PDA media culture and the highest growth diameter in high petro while had the lowest growth rate at all the All of these isolates were able to grow on crude petroleum as the sole source of carbon and energy when screened for hydrocarbon utilization. The growth diameter of fungus decreases with increasing petro concentration except sp in which the growth diameters of colonies were increased with increasing petro concentration 7 Thus I have reported the different types of fungus in my selected site i.e. Depot soil and Gram field soil. During the present work, heterotrophic fungal species related to a total of ten fungus were isolated from the petro polluted soil samples. These include Aspergillus Penicillium, Fusarium Mucor, Chaetomium, Cladosporium, Helminthosporium, Alternaria and Figure-1 Comparison between depot soil and gram field soil (pH, EC&Carbon)  International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Int. Res. J. Biological Sci. 29 and Penicillum species were present in dominant numbers. Our finding coincides with the who also isolated four strains from the contaminated soil. They were identified as Aspergillus sp. the most commonly encounte red genera of hydrocarbon degraders in oil contaminated tropical soils, which are in agreement with the present work Who isolated three fungi from total hydrocarbon contaminated soil and identified spergillus and Rhizopus sp11. The different result from our findings were obtained 12, who also isolated many fungal species that were able to degrade poly - cyclic aromatic hydrocarbons. The species isolated were Coniothyrium fuckelii, Gliocadium virens , Phialo-phora alba, ora hoffmannii, Scopulariopsis brumptii and along with genera Penicillium, which were similar to our finding. With the serial dilution plate Penicillium species were the richest The growth rate of each fungus had the highest growth diameter in low petro contaminated PDA media culture and Aspergillus sp had the highest growth diameter in high petro while Penicillium sp had the lowest growth rate at all the concentrations. All of these isolates were able to grow on crude petroleum as the sole source of carbon and energy when screened for hydrocarbon utilization. The growth diameter of fungus decreases with increasing petro concentration except Aspergillus in which the growth diameters of colonies were increased 7 . Thus I have reported the different types of fungus in my Gram field soil. During the present work, heterotrophic fungal species related to a total of ten fungus were isolated from the petro polluted soil samples. Penicillium, Fusarium , Rhizopus, Cladosporium, Morssonina, Curvularia, and Trichoderma species. Comparison between depot soil and gram field soil (pH, EC&Carbon)   !"  International Research Journal of Biological Sciences ________________________________________________ Vol. 3(11), 28-31, November (2014) International Science Congress Association Graphical comparison of soil analysis (N: P: K) Conclusion The present study was carried out at Manglia depot to compare the soil analysis of the two sites, Petro- polluted site and Gram field site. The comparison was done on the basis of different soil parameters including pH, EC, carbon concentration Nitrogen, Ph osphorus, and Potassium on the basis of the findings it can be concluded in the Petro- polluted site fungal growth is higher as compared to Gram field site. The soil samples examined for isolating fungi from petroleum hydrocarbon contaminated site in Mangli a diopot Indore M.P. by dilution technique were identified as Trichoderma, Fusarium, Alternaria, Curvularia Chaetomium, Morssonina, Helminthosporium.spp. Cladosporium.spp. , Mucor, Aspergillus, Penicillium, and Rhizopus species. Both sites reported si milar finding of some fungi like Alternaria, Curvularia, Mucor, Chaetomium, Petroleum depot soil as well as Gram field soil. Rhizopus Aspergillus,Cladosporium spp. can be used to remove of petroleum pollution in soil. So we can say that in the petro polluted site the growth of fungal diversity were higher due to more carbon concentration. Thus it may be concluded that the nature of Deuteromycetous fungi varies at di locations in the study. The behavior of Deuteromycetes is also different under different soil parameters. References 1.Das K. and Mukherjee A.K., Crude petroleum biodegradation efficiency of bacillus subtilish and Pseudomonas aeruginosa strains isolated from a petroleum- oil contaminated soil from North 2007)   #$%$&' International Research Journal of Biological Sciences ________________________________________________ International Science Congress Association Figure-2 Graphical comparison of soil analysis (N: P: K) The present study was carried out at Manglia depot to compare polluted site and Gram field site. The comparison was done on the basis of different soil parameters including pH, EC, carbon concentration Nitrogen, osphorus, and Potassium on the basis of the findings it can be polluted site fungal growth is higher as compared to Gram field site. The soil samples examined for isolating fungi from petroleum hydrocarbon contaminated site a diopot Indore M.P. by dilution technique were Alternaria, Curvularia , Chaetomium, Morssonina, Helminthosporium.spp. , Mucor, Aspergillus, Penicillium, and milar finding of some fungi like Fusarium, Chaetomium, species. In depot soil as well as Gram field soil. Penicillium, can be used to remove of petroleum pollution in soil. So we can say that in the petro polluted site the growth of fungal diversity were higher due to more carbon concentration. Thus it may be concluded that the nature of Deuteromycetous fungi varies at di fferent sites or locations in the study. The behavior of Deuteromycetes is also Crude petroleum -oil bacillus subtilish and strains isolated from a oil contaminated soil from North -East India, 2.Alexander M., Biodegradation and Bioremediation. Academic Press, New York, ( 3. 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In Cientifica, Centrode Tecnologia Mineral 55, (2006)9. Eja ME, Arikpo GE and Udo SM. potentials of fungal species is (' ')%$&'('%$&' International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Int. Res. J. Biological Sci. 30 Biodegradation and Bioremediation. Academic Press, New York, ( 1994) Obire O and Anyanwu EC. , Impact of various concentrations of crude oil on fungal populations of soil , International Journal Environmental Science and 218, ( 2009) Fungal metabolism of polycyclic aro - P ast, present and future applications in bioremediation , Journal of industrial Microbiology and Biotechnology 19(1), 324-333, (1997) Chaillan F, Fleche AL, Bury E, Phantavong Y, Crimount P, Saliot A and Oudot J. Identification and biodegradation potential of tropical aero bic hydro- degrading microorganism Research in 595, ( 2004) Akpoveta OV, Egharevba F and Medjor OW ., A pilot study on the hydrocarbon and its kinetics on kerosene International Journal of Environmental Akinyemi OJ and Adebusoye AS. , Hydrocarbon degrading Potentials of bacteria isolated from a Nigerian Bitumen (Tarsland) deposit , Nature and Reiche AP and Lemos JLS. , Estudo do potencial de degradacao de petroleo de linhagens de fungus isoladas : XIV Jornada de Iniciacao Cientifica, Centrode Tecnologia Mineral -CETEM/MCT Eja ME, Arikpo GE and Udo SM. The bioremediation potentials of fungal species is olated from soils polluted   !"  International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 3(11), 28-31, November (2014) Int. Res. J. Biological Sci. International Science Congress Association 31 by petroleum products in cross River University of Technology, Calabar, Nigeria, International Journal of Natural and Applied Science, 1, 15-20, (2006)10.Elisane ODS, Celia FCDR, Catia TDP, Ana VLS, Janaina FDMB, Susana JK and Carlos AVB.,Pre-screening of filamentous fungi isolated from a contaminated site in Southern Brazil for bioaugmentation purposes, African Journal of Biotechnology, 7, 1314-1317, (2008)11.Mancera-Lopez ME, Rodriguez-Casasola MT, Rios-Leal E, Esparza-Garcia F, Chavez-Gomez B, Rodriguez-Vazquez R and Barrera-Cortesa, J.Fungi and Bacteria isolated from Two Highly polluted Soils for Hydrocarbon Degradation, Acta Chimica Sloenica, 54 (1), 201-209, (2007)12.Ravelet C, Krivobok S, Sage L and Steiman R.,Biodegradation of Pyrene by Sediment Fungi, Chemosphere,40(5), 557-563, (2000)