Research Journal of Agriculture and Forestry Sciences __________________________________ ISSN 2320-6063 Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 13 Mycoflora of Some Spices from Dharwad, IndiaCh. Ramesh and Santoshkumar Jayagoudar Department of Botany, Karnatak University, Dharwad- 580003, INDIA Available online at: www.isca.in Received 20th July 2013, revised 9th August 2013, accepted 3rd September 2013Abstract Spices are most important agricultural commodities, commonly used to flavor the food preparations. Due to poor agriculture and storage practices spices get contaminated from field to fork level, among the contaminants fungi are the most predominated once, when they get the conducible atmosphere for their growth they grow and alters the spice quality and taste. The spices contaminated by fungi might cause the health hazards for humans. The fungal growth is mainly influenced by factors like Temperature and Relative Humidity. In the present investigation a total of 118 fungal species belongs to 44 genera were isolated out of which 18 species belonging to 08 genera of Zygomycetes, 07 species belonging to 02 genera of Ascomycets and 93 species belonging to 34 genera of Mitosporic fungi in the present investigation no Basidiomycetes were encountered. Among the isolates majority of the fungi belongs to the group Mitosporic fungi, than Zygomycetes and Ascomycetes group respectively, Genera like Rhizopus, Mucor and Absidia were dominant in Zygomycetes, Genus Chaetomium in Ascomycetes and in Mitosporic fungi genera like Aspergillus, Cladosporium, Curvularia, Penicillium, Alternaria and Fusarium are most predominant. Keywords: Mycoflora, contamination, spices, percentage of occurrence. Introduction Spices are important agricultural commodities, because of their taste and aroma they are commonly used to flavor the food preparations. Spices occupy a prominent place in the traditional and culinary practices all over the world1,2, coming to the history of spices, India is known as home of spices and one of the largest producer of spices due to poor agricultural and storage practices which affects the production and quality of spices3,4. Fungi are predominant contaminants of spices but most such mycopopulations are probably regarded as commensal residents on the plant that survived may be responsible for spoilage and degradation of spice quality by producing enzymes, mycotoxins and other secondary metabolites. Fungal growth of stored spices is mainly influenced by Temperature and Relative Humidity. Prevalence of seed spice mycoflora depending upon the field and storage conditions. The spices contaminated by fungi might cause the health hazards for humans. The spoilage of spice quality and mycotoxin production mainly influenced by type of fungi, food composition and also handling and storage practices. Several other reports have shown fungal contamination of different spices and other agricultural commodities. Francisco das Chagas O. et al reported a wide range of field and storage fungi from Black Pepper, White Pepper and Brazil nuts. Studies on spoilage mould and mycotoxigenic fungi from imported raw spices was screened by Qaher A. Mandeel. Farid M. Toma and Nareen Q. Faqi Abdulla10 Investigated of microbial status of some spices and crude herbal drugs. Analysis of commodity wise mycoflora, contamination status and mycotoxin magnitude in commercially important agricultural products reported by Vinod Kumar et al 11. In the present investigation a detailed survey of mycoflora of five selected spices from Dharwad region was under taken Viz; Pepper nigrum L. (Pepper), Coriandrum sativum L. (Coriander), Cuminum cyminum L. (Cumin), Elettaria cardomomum Maton.(Cardamom) and Cinnamomum tamela T. Nees and C.H. Eberm (Indian cassia). The screening of the mycoflora of spices from Dharwad region was done during the period of November 2009 to October 2012. Material and Methods Study area: Dharwad is selected as the study area for screening spice mycoflora, the area is located at 1544 North latitude 7499 East longitude in Karnataka state (India), because of the rich vegetation and conducive atmosphere for fungal growth it harbors different groups of fungi. The sites selected for the present investigation is Market Area and in and around Dharwad. Sample collections: The five spice samples Viz; Pepper nigrumL. (Pepper), Coriandrum sativum L. (Coriander), Cuminum cyminum L. (Cumin), Elettaria cardomomum Maton. (Cardamom) and Cinnamomum tamela T. Nees & C. H. Eberm (Indian cassia) were collected from Local market area Kirana merchant shops and spice sailors etc. on regularly monthly basis, from the chosen sites from during the period of November 2009 to October 2012. The selected spice samples of unknown variety was collected in sterile polyethylene bags separately from the chosen sites, during the collection field data Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 14 Viz; collection area, weight, type of sample, date of collection etc was carefully noted in the field note book. Collected samples were brought to the laboratory and used for further mycoflora analysis. Isolation of fungi from spices: Mycoflora of selected spices was isolated by using different isolation methods Viz. Serial Dilution Method12, Agar Plate Method13 and Standard Blotter Method14 for the isolation different media were used Viz. Potato Dextrose Agar (PDA), Czapek Dox Agar (CZA), Malt Extract Agar (MEA) Yeast Extract Agar (YEA) and Sabouraud Dextrose Agar (SDA), Chloramphenicol antibiotic is used to inhibit the Bacterial growth. Counting of Fungal Colonies: Fungal colonies start appearing on the inoculated plates after 24 hours white mycelium start to appear and in a couple of days, a full growth of the fungus takes place to form a colony. But in some cases, appearance of fungal colony takes many days as noticed in slow growing fungi. At the same time number of fungal colonies, color of the colony from both the sides and textures of the colony were noted. The incubated plates were observed under stereo-binocular microscope. To avoid loss of fungi which developed late, and which might be covered by fast growing forms, all plates in a series were checked frequently under stereo-binocular microscope for a period of two weeks. All plates observed for one month before discarding. Slide preparation, Photomicrography and Identification: Lactophenol with cotton blue stains was used for the slide preparation; the prepared slides were sealed with DPX mountant. The fungal illustrations was done based on Erma Camera Lucida drawings along with micrometric measurements by using 4X, 5X, 10X and 40X objectives 10X eye piece was used depending upon the fungal morphology and necessity. Photo micrographs of the prepared slides were taken by Carl Zeiss Imager M2 Model Microscope with Jenoptic Prog. Res. C5 attached Camera using 10X and 40X objectives and 10X eye piece. Identification of different fungi from five selected spices was done with help of slides prepared by direct mount from the culture; prepared slides were made permanent, with help of Erma Camera Lucida drawings along with micrometric measurements, Photomicrography and by referring fungal monographs and manuals such as Thom C. and Raper K.B.15, Raper K.B. and Thom C.16, Tandon R.N.17Subramanian C.V.18, Barnett H.L. and Hunter B.B.19, Domsch K.H. and Gams W.20Ainsworth G.C., Sparrow F.K. and Sussman A.S.21Ellis M.B.22Ellis M. B.23, Ellis M.B. and Ellis J.P.24Raper K.B. and Fennell D.I.25Bilgrami K.S., Jamaluddin S. and Rizwi M.A.26 Gilman J.C.27 Nagamani A. Kunwar I.K. and C. Manoharachary28Pande Alaka29 also research articles and other related literature fungal Identification and illustrations were made up to the Genera and Species level. All identified specimens are deposited, under the code number MASD (Mycoflora Analysis of Spices from Dharwad), in the Mycology Laboratory, Department of Botany, Karnatak University, Dharwad, Karnataka State, India. Presentation of data: In the presentation of data the terms “Percentage of fungal occurrence” were used, The term percentage of fungal occurrence is used to denote, Results and DiscussionThe present study reveals the isolation of 118 fungal species belongs to 44 genera, out of which 18 species belonging to 08 genera of Zygomycetes, 07 species belonging to 02 genera of Ascomycets and 93 species belonging to 34 genera of Mitosporic fungi. Among the isolates majority of the fungi belongs to the Mitosporic fungi group, than Zygomycetes and Ascomycetes group respectively (table 1). Table-1 Different groups of fungi isolated from selected SpicesGroups Genera Species Zygomycetes 08 18 Ascomycetes 02 07 Mitosporic fungi 34 93 Total 44 118 The predominant fungal genera were Aspergillus, Cladosporium, Curvularia, Penicillium, Alternaria and Fusarium in Mitosporic fungi, in Ascomycets genus Chaetomium is dominant and in Zygomycetes genera like Rhizopus, Mucor and Absidia were dominant. Among the five selected spices samples Cumin samples shows highest number of fungal species isolates belongs to different groups Viz. Zygomycetes (14), Ascomycets (05), Mitosporic fungi (65); from Coriander Zygomycetes (10), Ascomycets (05), Mitosporic fungi (61) isolates; Pepper Zygomycetes (11), Ascomycets (04), Mitosporic fungi (45); Cardamom shows Zygomycetes (09), Ascomycets (01), Mitosporic fungi (38) and Indian cassia samples shows the least number of fungal isolates Zygomycetes (06), Ascomycets (04), Mitosporic fungi (25) respectively (table 2 and figure 1). Isolation of Fungi with respect to the different isolation methods Serial dilution method shows highest number of fungal species isolates belongs to different fungal groups viz. Zygomycetes (16), Ascomycets (05), Mitosporic fungi (78) species from Agar Plate Method Zygomycetes (14), Ascomycets (04), Mitosporic fungi (63) species and Standard Blotter Method Zygomycetes (10), Ascomycets (07), Mitosporic fungi (40) species were isolated respectively (figure 2).Among thefungal media PDA media shows highest number of isolates in total (84) species isolates; MEA shows total (66) species isolates; SDA shows Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 15 total (58) species isolates; CZA shows total (29) species isolates and YEA shows total (16) species isolates respectively (figure 3). The percentage of fungal occurrence reveals predominantly occurring fungal genera like Rhizopus (2.799%), Mucor (1.806%) and Absidia (1.320%) in Zygomycetes, Genus Chaetomium shows (2.849%) occurrence in Ascomycetes and in Mitosporic fungigenera like Aspergillus (25.434%),Cladosporium (8.825%), Curvularia (8.756%), Penicillium (7.157%), Alternaria (5.976%) and Fusarium (5.906%) are most predominant genera. The highest percentage of occurrence was recorded in the genus Aspergillus (25.434%) and the lowest percentage of occurrence was observed in the genus Rhynchophoma (0.138%) and other genera shows percentage of occurrence ranges between (0.138% to 25.434%) (figure 4). The other workers screened mycobiota of different spices and reported the predominant fungal genera viz. Aspergillus,Eurotium and Fusarium from Capsicum powder samples by Santos L et al30. Abdulkadir E. Elshafie et al31 isolated twenty fungal species out of which species like Aspergillus flavus, Aspergillus niger, Penicillium Spp., Rhizopus spp. and Syncephalastrum Spp. were predominately isolated ; Abou Donia M. A 32screened 303 samples representing different types of spices and medicinal plants from Egypt and reported genera like Aspergillus,Fusarium,Cladosporium,Penicillium, Mucor Absidia and Rhizoctonia were dominant among the isolates. Hashem Mohamed et al33 examined fifteen spices for their mould profile and isolated a total of 57 species. I. A. E- Kady et al34 isolated 81 species belongs to 38 genera of fungi from 24 different of spices, collected from Egypt. Ahene, R.E.et al35 screened fungal and bacterial contaminants of six spices and spice products in Ghana. Maetrese Arianne J. Beley et al36 isolated fungal species from soy milk products and also analyzed the presence of aflatoxin level by using Enzyme-Linked Immunosorbent Assay (ELISA) method. Abedi-Tizaki Mostafa et al37 reviewed rapid detection methods for analysis of Fungi and Mycotoxins in agriculture products. Figure-1 Different species of fungi isolated from five selected spices Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 16 Table-2 Different fungal species isolated from five Spices from Dharwad Sl. No. Name of the fungi Spices ZYGOMYCETES Pep. Cori. Cumin Cardo. In. Cassia 1 Absidia butleri Lendner + + + - - 2 Absidia lichtheinsmi Lendner + - + + - 3 Choanephora cucurbitarum Thaxter - + + + - 4 Circinella simplex Tiegh. + + - - + 5 Cunninghamella elegans Lendner + + + - - 6 Mucor bacilliformis Hesselt. - + + + - 7 Mucor fragalis Bainier + - + + + 8 Mucor hiemalis Wehmer + + + - - 9 Mucor javanicus Wehmer - - + + + 10 Mucor rouxianus Wehmer + + - - - 11 Mucor varians Povah + - - + - 12 Mycotypha indica P.M. Kirk & Benny - + + - - 13 Rhizopus arrhizus Fisher + + + - - 14 Rhizopus nigricans Ehrenb - - + + - 15 Rhizopus nigricans Ehrenbvar. minutus Chaudhuri- - + + + 16 Rhizopus nodosus Namyslowski + - - + - 17 Rhizopus oryzae Went et Gerlings - + + - + 18 Syncephalastrum racemosum Cohn ex J. Schrot. + - + - + ASCOMYCETES 19 Chaetomium amberpetense Rama Rao & Ram Reddy- + + - + 20 Chaetomium atrobrunneum Udagawa & Takada- + - + - 21 Chaetomium caprophilum Narendra & VG Rao + - + - + 22 Chaetomium mollicellum L. M. Ames. + - + - + 23 Chaetomium spirale Zopf. + + - - + 24 Chaetomium subterraneum Swift & Povah - + + - - 25 Emericella nidulans Vuill. + + + - - MITOSPORIC FUNGI 26 Aegerita candida Pers. - + + + - 27 Alternaria amaranthi van Hook + + - + - 28 Alternaria burnsii Uppal, Patel & Kamat - + + - + 29 Alternaria carthami Chowdhury+ - + - + 30 Alternaria Citri Ellis & Pierce+ - + + - 31 Alternaria cyamopsidis Rangaswami & Venkatarao - + - + - 32 Alternaria gomphernae Togashi- + + + - 33 Alternaria longipes Mason- - + + + 34 Alternaria palandui Ayyangar + + + - - 35 Alternaria ricini Hansford - + + + - 36 Alternaria tenuissima Wiltshire + - + - - 37 Aspergillus alliaceus Thom & Church - + + - + 38 Aspergillus ambiguous Sappa.+ + + - - 39 Aspergillus awamori Nakazawa - + - + + 40 Aspergillus caespitopsis Raper &Thom + - + - + 41 Aspergillus candidus Link. + + + - - 42 Aspergillus chevalieri Thom & Church - + - + + 43 Aspergillus deflectus Fenell & Raper + - + - + 44 Aspergillus flavus Link. + + - - - 45 Aspergillus funiculosis Smith - - - + + 46 Aspergillus heteromorphus Batista & Maia + + + - - Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 17 47 Aspergillus kanagawaensis Nehira- + - + + 48 Aspergillus microcysticus Sappa. - + + + - 49 Aspergillus montevidensis Talice & Mackinnon + + - + - 50 Aspergillus ochraceus Wilhelm + + + - - 51 Aspergillus parasiticus Speare - - + + - 52 Aspergillus petrakii Voros + - - + + 53 Aspergillus puniceus Kwon & Fenell - + + - - 54 Aspergillus repens De Bary + - + + - 55 Aspergillus speluneus Raper & Fennell+ - - - + 56 Aspergillus spinuloses Warcup+ - + + - 57 Aspergillus tericola var. indicus N. Comb.- + - + + 58 Aspergillus terricola Marchal- + + + - 59 Aspergillus ustus Thom & Church + + - + - 60 Aspergillus versicolor Tiraboschi+ - + - + 61 Aurobasidium olae Subram. Comb. nov. - + + - - 62 Bipolaris tetramera Shoemaker - + + + - 63 Cladosporium cladosporioides de Vries+ + - - + 64 Cladosporium fulvum Cooke. - - + + - 65 Cladosporium herbarum Link ex Fries + + + - - 66 Cladosporium oxysporum Berk. & M.A. Curtis + + + + - 67 Cladosporium variabile de Vries+ + + - - 68 Cochliobolus australiensis Alcorn - + + - - 69 Curvularia inaequalis Boedijn + - + - - 70 Curvularia indica Subram. + + - + - 71 Curvularia lunata Boedijn - - + + + 72 Curvularia maculans Boedijn + - + - - 73 Curvularia pallescens Boedijn - + + - - 74 Curvularia prasadii Mathur R.L. &B. L. - + - - + 75 Curvularia senegalensis Subram. + - + - + 76 Exosporina laricis Oudem. + - + - - 77 Fusariella intermedia Mouchacca & Nicot - + + - - 78 Fusarium merismoids Corda + + - - - 79 Fusarium nivale Cesati - + + + - 80 Fusarium oxysporum Schl. ex Fries + - + - + 81 Fusarium poae Wr. + + - - - 82 Fusarium udum Butler - + + - - 83 Fusicladiella melaena Hughes - + + - - 84 Fusidium aureum Link. - + - + - 85 Geotrichum candidum Link. ex. Sacc. + - + - - 86 Gliocladium penicillioides Corda - + + + - 87 Gliocladium roseum Bainier + - + - - 88 Memnoniella echinata Galloway - + + - - 89 Memnoniella levispora Subram. + + - - - 90 Monodictys fluctuate M. B. Ellis - + + - - 91 Monosporium acuminatum var. terrestre Sacc. - + + - - 92 Nigrospora oryzae Petch - - + - + 93 Nigrospora sphaerica Mason + - + - - 94 Paecilomyces varioti Baineir - + - + - 95 Papulaspora irregularis Hotson - + + - - 96 Penicillium brefeldianum Dodge + - + - - 97 Penicillium chermesinum Biourge + - - + - 98 Penicillium cyaneum Biourge - + - - + 99 Penicillium decumbens Thom + + + - - Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 18 100 Penicillium janthinellum Biourge - + - + - 101 Penicillium nigricans Bainier ex Thom + - - + + 102 Penicillium rubrum Stoll - - + + + 103 Periconia cookei Manson & Ellis - + + - - 104 Phragmostilbe linderi Subram. + - + - - 105 Rhynchosporium secalis Davis - + + - - 106 Sclerococcum sphaerale Fr. - + + - - 107 Sporidesmium vagum C.G. Nees & T.F.L. Nees ex Link + + - - - 108 Stachybotrys chartarum Hughes - + + - + 109 Stachybotrys pulchra Speg. - + + + - 110 Trichoderma koningii Oudem + + + - - 111 Trichoderma longibrachiatum Rifai - - + - + 112 Trichoderma viride Pers. - + + + - 113 Veronaea caprophola M.B. Ellis comb. nov.- + + - - 114 Verticillium dahliae Klebahn + - + + - 115 Verticillium luteo-album Subram. + + - - - 116 Wardomyces anomala Brooks & Hansf. + - - + - COELOMYCETES 117 Phoma herbarum Cooke - + + - - 118 Rhynchophoma raduloides Karst. - + - + - TOTAL 60 76 84 48 35 + = Present, - = Absent., Pep. - Piper nigram Linn. (Pepper), Cori. – Coriandrum sativum Linn. (Coriander) , Cumin - Cuminum cyminum Linn. (Cumin), Cardo. - Elettaria cardomomum Maton. (Cardamom), In. Cassia. - Cinnamomum tamela T. Nees & Eberm (Indian Cassia) Figure-2 Isolation of different classes of fungi by using different methods Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 19 Figure-3 Distribution of different classes of fungi in different media Conclusion Spices are most important agricultural products because of their taste and aroma they are commonly used to flavor the food preparations. India is known as home of spices and one of the largest producer spices and they occupy a prominent place in traditional culinary practices, because of poor agriculture, storage and handling practices spices become contaminated at each level. When the contaminated mycoflora get conducive atmosphere for their growth; they grow and alter the quality and taste of the spices by secreting enzymes, toxins and other secondary metabolites during their growth and development. The present investigation resulted in the isolation and identification of 118 species of fungi belongs to 44 genera out of which 18 species belonging to 08 genera of Zygomycetes, 07 species belonging to 02 genera of Ascomycets and 93 species belonging to 34 genera of Mitosporic fungi. In the present screening no basidiomycetes were encountered. Among the isolates majority of the fungi belongs to the Mitosporic fungi group, than Zygomycetes and Ascomycetes group respectively. Acknowledgements My sincere thanks to UGC New Delhi, for financial assistance grant in the form of UGC MRP Project Fellowship and RFSMS Fellowship and also to the Director USIC, Karnatak University, Dharwad for providing the necessary instrumentation facilities and to the Chairman Department of Botany, Karnatak University, Dharwad for providing the Laboratory facilities. Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 20 Figure-4 Percentage of fungal occurrence Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 21 References 1.Atanda O.O., Akpan I. and Oluwafemi F., The potential of some Spice essential oil in the control of Aspergillus parasiticus CFR223 and aflatoxin production, Food control, 18, 601-607 (2007) 2.Oluwafemi F., Correlation between dietary Aflatoxins and human male infertility Ph.D. Thesis university of Benin, Benin City Nigeria (2000) 3.Sumanth G.T., Bhagawan M.W. and Surendra R.S., Incidence of mycoflora from seeds of Indian main spices, African Journal of Agriculture Research, 5(22), 3122-3125 (2010)4.Hedawoo and Rajana G., Chakranarayan Isolation of fungal species from the seeds of some Indian spices, Biosci. Biotech. Res. Comm.,4(2), 208-210 (2011)5.Misra Nisha, Influence of temperature and relative humidity on fungal flora of some spices in storage Zeitschrift für Lebensmittel-Untersuchung und Forschung, 172, 30-31 (1981)6.Singh Bhavana, Gitansh and Bhadauria Seema Prevalence of seed mycoflora from different seeds of spices under field and storage conditions of Agra region, Asian J. of plant Sci. and Res., 3(2), 93-98 (2013) 7.Abdulkadir E. Elshafie, Tahiya A. Al-Rashdi, Saif. N. Al-Bahry and Charles S. Bakheit Fungi and aflatoxins associated with spices in the Sultanate of Omen Mycopathologia, 155, 155-160 (2002)8.Francisco das Chagas O. Freire, Zofia Kozakiewicz & R. Russell M. Paterson Mycoflora and mycotoxins in Brazillian Black pepper, White pepper and Brazil nuts Mycopathologia,149, 13-19 (2000) 9.Qaher A., Mandeel Fungal contamination of some imported spices, Mycopathologia,59, 291-298 (2005)10.Farid M. Toma and Nareen Q. Faqi Abdulla Isolation and identification of fungi from Spices and medicinal plants, Research Journal of Environmental and Earth Sciences,5(3), 131-138 (2013) 11.Vinod Kumar, Basu M.S., Rajendran T.P., Mycotoxin research and mycoflora in some commercially important agricultural commodities, Crop Protection, 27, 891-905 (2008)12.Waksman S.A., Principles of Soil Microbiology, Williams and Wilkins Company, Baltimore, 897 (1927) 13.Muskette A.E., Techniques for the examination of seeds for the presence of seed born fungi, Trans. Brit. Mycol. Soc., 30, 74-83 (1948) 14.De Tempe J., The blotter methods of seed health testing proc, Int. seed test assoc., 28, 133-151 (1953) 15.Thom C. and Raper K.B., A Manual of the Aspergilli, Williams and Wilkins Company, Baltimore, 373 (1945) 16.Raper K.B. and Thom C., A Manual of the Penicillia, Williams and Wilkins, Baltimare, 850 (1948) 17.Tandon R.N., Mucorales of India, I.C.A.R. Publication, New Delhi, 120 (1968)18.Subramanian C.V., Hyphomycetes ICAR publication New Delhi edn., 930 (1971)19.Barnett H.L. and Hunter B.B., Illustrated genera of Imperfect fungi, III. Edn. Burgess Publishing Company, Minneapolis, Minnesota, 241 (1972) 20.Domsch K.H. and Gams W., Fungi in agricultural soil Longman group Ltd. London, 290 (1972) 21.Ainsworth G.C., Sparrow F.K. and Sussman A.S., The fungi- An advanced treatise, A taxonomic review with keys: Ascomycetes and Fungi imperfecti, Academic Press, IV(A), 621 (1973) 22.Ellis M.B., Dematiaceous Hyphomycetes. Common wealth Mycological Institute, Kew, England, 608 (1971) 23.Ellis M.B., More Dematiaceous Hyphomycetes. Common wealth Mycological Institute, Kew, England, 507 (1976) 24.Ellis M.B. and Ellis J.P., Microfungi on Land Plants [An Identification Hand Book], Croom Helm Ltd., Sydney, Australia, 818 (1985)25.Raper K.B. and Fennell D.I., The Genus Aspergillus, Robert E. Krieger Publishing Company, New York, 583(1965) 26.Bilgrami K.S., Jamaluddin S. and Rizwi M.A., Fungi of India, part I, Today and tomorrow’s Printer and Publishers, New Delhi, 467 (1981)27.Gilman J.C., A manual of soil fungi 2nd Indian edn. Biotech book publication Delhi, 382 (2001) 28.Nagamani A., Kunwar I.K. and Manoharachary C., Hand Book of Soil Fungi, I.K. International Pvt. Ltd. 477 (2006) 29.Pande Alaka, Ascomycetes of Peninsular India. Scientific Publishers Jodpur (India), 368 (2008) 30.Santos L., Marín S., Mateo E.M., Gil-Serna J., Valle-Algarra F.M., Patino B., Ramos A.J., Mycobiota and co- occurrence of mycotoxins in Capsicum powder, International Journal of Food Microbiology,151, 270-276 (2011)31.Abdulkadir E. Elshafie, Tahiya A. Al-Rashdi, Saif N. Al-Bahry and Charles S., Bakheit Fungi and aflatoxins associated with spices in the Sultanate of Omen, Mycopathologia,155, 155-160 (2002)32.Abou Donia M.A., Microbiological quality and aflatoxicogenesis of Egyptian spices and medicinal plants Global Veterinaria,2(4), 175-181 (2008) Research Journal of Agriculture and Forestry Sciences _______________________________________________ ISSN 2320-6063Vol. 1(8), 13-22, September (2013) Res. J. Agriculture and Forestry Sci. International Science Congress Association 22 33.Hashem Mohamed, Alamri Saad Contamination of common spices in Saudi Arabia markets with potential mycotoxin-producing fungi, Saudi Journal of Biological Sciences,17, 167–175 (2010)34.I.A. E- Kady, S.S. Mohamed E-Maraghy and Eman Mostafa M., Contribution of the Mesophilic fungi of different spices in Egypt, Mycopathologia,120, 93-101 (1992)35.Ahene R.E., Odamtten G.T. and Owusu E., Fungal and bacterial contaminants of six spices and spice products in Ghana, African Journal of Environmental Science and Technology, 5(9), 633-640 (2011)36.Maetrese Arianne J. Beley, Franco G. Teves, Ma. Reina Suzette B., Madamba Isolation of Fungal Species and Detection of Aflatoxin from Soy Milk Products using ELISA method, Int. Res. J. Biological Sci,2(5), 45-48 (2013)37.Abedi-Tizaki Mostafa, Askari Armin, Paknezhad Hamid and Abedi Mohammad Reza Rapid Detection Methods for Analysis of Fungi and Mycotoxins in Agriculture Products, Res.J.Recent Sci.,1(7), 90-98 (2012)