Studies On Strepto Mycetes Of Qualubia Governorati :

Ahmed Helmi Mohmmed

The present thesis aimed at the isolation, characterization, andtaxonomic identification of Streptomycetes of some soils of Qalubiagovernorate, and establishing a culture collection of theseStreptomycetes.Using starch nitrate agar and applying dilution teclmique, 154Streptomyces isolates were obtained from five soil samples covering thesoil types of Qalubia governorate. All isolates proved to beStreptomycetes.Using the color wheels of Treesner and Backus (1963), theobtained isolates were differentiated into 4 color sections:1. Yellow section that contains 29 isolates.2. Red section that contains 26 isolates.3. Gray section that contains 93 isolates and which were furtherdifferentiated into two color subsections:Sub - section (A) of red gray Streptomycetes that contains 66isolates.Sub - section (B) of light gray Streptomycetes that contains 27isolates.4. Blue section that contains 6 isolates.The isolates are further differentiated into groups according to thecolor of aerial and substrate mycelium, pigmentaion, morphology ofspore chains, and ornamentation of spores. As a result of this study theexperimented isolates were differentiated into 24 groups. Representativeisolates are subjected to detailed taxonomic studies as recommended bythe newest determinative manuals of Streptomycetes i.e. Bergey’sManual of Systematic Bacteriology Volume 4, (1989).218-----._---Yellow section: The 29 isolates of this section were differentiatedinto 4 groups as following:Group (1): This group includes five isolates of which isolate Y-IIwas chosen as the representative one. This isolate produces short straightchains of spores, short cylindrical spores with abrupt ends, spores are ofsmooth surface. It produces pale yellow aerial mycelium but light yellowgreen substrate mycelium. The green pigmented were found to be pHsensitive (pink -+ green). No melanin pigments are produced. Growthwas sensitive to some inhibitors, and some antibiotics. Utilized a wide setof carbon and nitrogen sources. Mesophile and neutrophile. Producesantimicrobial substances against gram (+ve) bacteria, yeasts, and fungi.Exhibits well expressed lipolytic activities. Degrades a wide set oforganic complex compounds, This group is identified as Streptomycesstreptomycini (Krassilnikov, 1970).Group (2): This group includes five isolates of which Y-18 waschosen as the representative one. It produces very long straight chains ofspores, arranged in tufts. Spores are cylindrical with abrupt ends andsmooth surface. Aerial mycelium is pale yellow but substrate mycelium islight brown. Produces pH sensitive green pigments. Melanin negative.Growth was sensitive to some inhibitors and antibiotics. Utilizes somecarbon and nitrogen sources. Mesophile and neutrophile. Producesantimicrobial substances that inhibit the growth of gram (+ve) bacteria,yeasts and fungi, Exhibits well express lipolytic, keratinolytic, cellolytic,and chitinolytic activities. Degrades some organic complex compounds. Itis identified as Streptomyces globisporus (Krassilnikov, 1941).Group (3): This group includes nine isolates of which Y-9 waschosen as the representative one. Produces long straight chains of sporeswhich may be slightly wavy. Spores are short cylinders with smoothsurface. Aerial and substrate mycelia are pale yellow. No diffusible219----._---- ---_ ..pigments. Melanin negative. Growth is sensitive to some inhibitors andantibiotics. Utilizes a set of carbon and nitrogen sources. Mesophile andneutrophile. Produces some antimicrobial substances against gram (+ve)bacteria, yeasts, and fungi, Exhibits well lipolytic, keratinolytic,pectinolytic, and chitinolytic activities. Degrades some organic complexcompounds. It is identified as Streptomyces vulgaris (Nikitina et al,1961).Group (4): This group includes ten isolates of which Y-20 waschosen as the representative one. It produces long straight chains ofspores that arranged in tufts, and long cylindrical spores with abrupt endsand interposal pads, and smooth surface. Aerial and substrate myceliaare pale yellow. No diffusible pigments. Melanin negative. Growth issensitive to some inhibitors and antibiotics. Utilizes a set of carbon andnitrogen sources. Mesophile and neutrophile. Produces antimicrobialsubstances against gram (-ve) bacteria, gram (+ve) bacteria; yeasts, andfungi. Exhibits well lipolytic, keratinolytic, cellulolytic, pectinolytic, andchitinolytic activities. Degrades some organic complex compounds. Thisgroup was found to be dissimilar to any described species of the yellowStreptomyces and so it is declared as a new species to which the nameStreptomyces albolongus (Hussein and Mohammed, 2005) was coined.Red section: The 26 isolates of this section are differentiated into 4groups as following:Group 5: This group includes eight isolates of which isolate R-2was chosen as the representative one. This isolate produces, moderate inlength straight chains of spores, long cylindrical spores with abrupt endsand smooth surface. It produces pale orange yellow aerial mycelimn butlight brown substrate’ mycelium. No diffusible pigments. Melaninnegative. Growth is inhibited by some inhibitors and antibiotics. Utilizes.. 220a set of carbon and -nitrogen sources. Mesophile and neutrophile.Produces antimicrobial substances, against gram (+ve) bacteria, yeastsand fungi, Exhibits well lipolytic, keratinolytic, cellulolytic activities.Degrades some organic complex compounds, This isolate is identified asStreptomyces gilvus (Krassilnikov, 1970).Group 6: This group includes five isolate of which R-3 waschosen as the representative one. It produces very long straight chains ofspores which are with well extraordinarily long cylindrical spores withsmooth surface. Aerial mycelium is grayish yellowish pink but substratemycelium is light brown. No diffusible pigments. Melanin negative.Growth was affected by some inhibitors and antibiotics. Utilizes somecarbon and nitrogen sources. Mesophile and neutrophile. Producesantimicrobial substances against gram (+ve) bacteria, yeasts and fungi.Exhibits lipolytic, keratinolytic, pectinolytic, and chitinolytic activities.Degrades some organic complex compounds. It is identified asStreptomyces stramineus (Hussein and Konsoh, 1981).Group 7: This includes four isolates of which R-41 was chosen asthe representative one. It produce primitive loose spiral chains of spores,Spores are long with tapering ends and smooth surface. Aerial myceliumis grayish yellowish pink but substrate mycelium is moderate yellowishpink. No diffusible pigments. Melanin negative. Growth is sensitive tosome inhibitors and ’antibiotics. Utilizes some carbon and nitrogensources. Mesophile and neutrophile. Produces antimicrobial substancesagainst gram (+ve) bacteria, yeasts and fungi, Exhibits well lipolytic,keratinolytic, cellulolytic, and chitinolytic activities. Degrades someorganic complex compounds, It is identified as the new speciesStreptomyces roseobrunneus n.sp. (Hussein and Mohammed, 2005).Group 8: This group includes nme isolates of which R-5 waschosen as the representative one. It produces long straight chains ofspores that ends with quite open loops or hooks in the form of tufts.Spores are long cylinders with abrupt ends and smooth surface. Aerialmycelium is moderate yellowish pink but substrate mycelium is paleyellow. No diffusible pigments. Melanin positive. Growth is sensitive tosome inhibitors and antibiotics. Utilizes some carbon and nitrogensources. Mesophile and neutrophile. Produces antimicrobial substancesagainst gram (-ve), gram (+ve) bacteria, yeasts and fungi. Exhibits welllipolytic and chitinolytic activities. Degrades some organic complexcompounds. It is identified as Streptomyces lavendulae (William andHenrici, 1948).Gray section: The 93 isolates of this section are differentiated into2 colored subsections as follows:Sub-section (A): Red gray isolates that contains 66 isolates, andthat are differentiated into 9 groups according to the ornamentation ofspores, pigmentation and morphology of spore chains.Group 9: This group includes three isolates of which isolateG-100 was chosen as the representative one. It produces long loose spiralspore chains that consist of 3-6 turns. Spores are cylindrical with roundedends and smooth surface. Aerial mycelium is light grayish reddish brownbut substrate mycelium is grayish yellow. It does not produce pHsensitive pigments. Melanin negative. Growth is sensitive to someinhibitors and antibiotics. Utilizes some carbon and nitrogen sources.Mesophile and neutrophile. Produces antimicrobial substances that inhibitgram (+ve) bacteria and yeasts. Exhibits well lipolytic, keratinolytic,cellolytic and pectinolytic activities. Degrades some organic complex222compounds, It is identified as Streptomyces humldus (Nakazawa andShibata, 1958).Group 10: This group includes seven isolates of which G-121 waschosen as the representative one. it produces short stalked spore chainswith 1-2 turns. Spores are short cylindrical with smooth surface. Aerialmycelium is grayish yellowish pink but substrate mycelium is lightorange yellow, the brown diffusible pigments are found to be pHsensitive (brown ~ yellow). No melanin pigments are produced. Growthis sensitive to some inhibitors and antibiotics. Utilizes a wide set ofcarbon and nitrogen sources. Mesophile and neutrophile. Producesantimicrobial substances against gram (-ve), gram (+ve) bacteria, yeastsand fungi. Exhibits well expressed lipolytic, keratinolytic, cellulolytic,pectinolytic, keratinolytic, cellulolytic, pectinolytic, and chitinolyticactivities. Degrades a wide set of organic complex compounds. Thisgroup is identified as Streptomyces giivocinereus (Krassilnikov, 1970).Group 11: This group includes twenty one isolate of which G-23was chosen as the representative one. It produces straight spore chainswith straight ends, spores are long cylindrical with smooth surface. Aerialmycelium is light grayish reddish brown but substrate mycelium is lightbrown. It does not produce pH sensitive pigments. Melaninnegative. Growth is sensitive to some inhibitors and antibiotics. Utilizessome carbon and nitrogen sources. Mesophile and neutrophile. Producesantimicrobial substance against gram (+ve) bacteria and yeasts. Exhibitswell lipolytic, keratinolytic, pectinolytic, and chitinolytic activities.Degrades some organic complex compounds. It is identified asStreptomyces misakiensis (Nakamura, 1961).Group 12: This group includes five isolates of whichG-I07 was chosen as the representative one. It produces loose spiralspore chains, consist of 2-4 turns. Spore are long cylindrical with rounded223-- ---.ends and smooth surface. Aerial and substrate mycelia are brownish gray,No diffusible pigments. Melanin negative. Growth is sensitive to someinhibitors and antibiotics. Utilizes some carbon and nitrogen sources.Mesophile and neutrophile. Produces antimicrobial substances againstonly yeasts. Exhibits lipolytic, Keratinolytic, cellulolytic, pectinolytic,and chitinolytic activities. Degrades some organic complex compounds. Itis identified as Streptomyces capuensis (Krassilnikov, 1970).Group 13: This b’fOUP includes twelve isolates of which G-49 waschosen as the representative one. It produces well developed loose spiralspore chains. Spores are short cylindrical with rounded ends, sporesurface is smooth. Aerial mycelium is light gray but substrate myceliumis pale blue. No diffusible pigments. Melanin negative. Growth issensitive to some inhibitors and antibiotics. Utilizes some carbon andnitrogen sources. Mesophile and neutrophile. It does not produce anyantimicrobial substances against gram (we), gram (+ve) bacteria, yeasts,or fungi. Exhibits lipolytic, keratinolytic, cellulolytic, pectinolytic, andchitinolytic activities. Degrades some organic complex compounds. It isidentified as Streptomyces rubrocyaneus (Hussein, 1964).Group 14: This group includes one isolate GNG5. It producesshort spore chain in the form of somewhat close spiral. Spores are shortcylindrical, with rounded ends, spore surface is with fine short acutespines. Aerial mycelium is moderate gray but substrate mycelium is lightolive brown. No diffusible pigments. Melanin negative. Growth issensitive to some inhibitors and antibiotics. Utilizes some carbon andnitrogen sources. Mesophile and neutrophile. Produces antimicrobialsubstances against only yeasts. Exhibits well lipolytic, keratinolyticcellulolytic, pectinolytic and chitinolytic activities. Degrades someorganic complex compounds. It is identified as Streptomyces toyocaensls(Nishinwra et ai, 1950).. --_._-· Group 15: This hTfOUP includes nine isolates of which G-69 waschosen as the representative one. It produces short chains of spore in theform of hooks or loops spores are elongated cylinders with abrupt ends,spore surface is covered with fine acute spines. Aerial mycelium is lightgray but substrate mycelium is pale yellow green, No diffusiblepigments. Melanin negative. Growth is sensitive to some inhibitors andantibiotics. Utilizes some carbon and nitrogen sources. Mesophile andneutrophile. Produces some antimicrobial substances against only yeasts.Exhibits well lipolytic, keratinolytic, pectinolytic and chitinolyticactivities. Degrades some organic complex compounds. It is identified asStreptomyces greseorubens (Preobrajenskaya et al., 1968).Group 16: This hTfOUP includes one isolate G-64. It produce shortloose spiral spore chains in the form of tufts. Spores are short cylinderswith sounded ends. Spore surface is densely covered with long broadhairs. Aerial mycelium is pale blue but substrate mycelium is darkgreenish gray. No diffusible pigments. Melanin negative. Growth issensitive by some inhibitors and antibiotics. Utilizes set of carbon andnitrogen sources. Mesophile and neutrophile. Produces some antibioticsagainst (+ve) bacteria and fungi. Exhibits well lipolytic, keratinolytic,cellulolytic, pectinolytic and chitinolytic activities. Degrades someorganic complex compounds. It is identified as the new speciesStreptomyces cyanotrichosporus n.sp. (Hussein and Mohamed, 2005).Group 17: This group includes seven chosen of which G-7 waschosen as the representative one. It produces loose spiral spore chainsconsists of 1-2 turns, spirals are stalked. Spores are very long cylindricalwith abrupt ends, spore surface is hairy. Aerial mycelium is dark gray butsubstrate mycelium is pale yellow. No diffusible pigments. Melaninnegative. Growth is affected by some inhibitors and antibiotics. Utilizessome carbon and nitrogen sources. Mesophile and neutrophile. Produces225._----some antimicrobial substances against gram (+ve) bacteria and yeasts.Exhibits well lipolytic, keratinolytic, cellulolytic, pectinolytic andchitinolytic activities. Degrades some organic complex compounds. It isidentified as Streptomyces sporoplllosus (Krassilnikov, 1970).Subsection (B)This subsection includes 27 isolates and differentiated into 6groups depending on the morphology of spore chains and color of aerialmycelium.Group (18): This group includes six isolates of which G-115 waschosen as the representative one. It produces open spiral spore chains inthe forms of hooks. Spores are of big size in the form of short cylindersand smooth surface. Aerial mycelium is light gray but substrate myceliumis light yellow. No diffusible pigments. Melanin negative. Growth issensitive to some inhibitors and antibiotics. Utilizes some carbon andnitrogen sources. Mesophile and neutrophile. Produces antimicrobialsubstances against only yeasts. Exhibits well lipolytic, keratinolyticcellulolytic, pectinolytic and chitinolytic activities. Degrades someorganic complex compound. It is identified as Streptomyces canofumeus(Krassilnikov 1970).Group (19): This group includes four isolates of which G -101was chosen as the representative one. This isolate short primitive chainsof spores in the form of hooks or loops. Spores are in the form of shortcylinders with abrupt ends, spore surface is smooth. Aerial mycelium islight gray but substrate mycelium is light yellow. Produces light yellowdiffusible pigments. Melanin production is positive. Growth is sensitiveto some inhibitors and antibiotics. Utilizes some carbon and nitrogensources. Mesophile and neutrophile. Produce antimicrobial substancesagainst only yeasts. Exhibits well lipolytic, keratinolytic, cellulolytic.226pectinolytic and chitinolytic activities. Degrades some organic complexcompounds. It is identified as Streptomyces cinereous (Krassilnikov,1970).Group (20): This group includes three isolates of which G-72 waschosen as the representative one. It produces stalked spiral chains ofspores, consisting of 2-3 turns. Spores are cylindrical with abrupt endsand smooth surface. Aerial mycelium is medium gray but substrate islight olive brown. No diffusible pigments. Melanin negative. Growth issensitive to some inhibitors and antibiotics. Utilizes some carbon andnitrogen sources. Mesophile and neutrophile. Produces antimicrobialsubstances against gram (+ve) bacteria and yeasts. Exhibit well lipolytic,keratinolytic, pectinolytic and chitinolytic activities. Degrades someorganic complex compounds. It is identified as Streptomycesanlanoslnicus. (Thieman and Beretta, 1966).Group (21): This group includes eight isolates of which GNG8was chosen as the representative one. It produces well developed spiralspore chains that have 3-4 turns, spores are oblong, with attenuated ends,spore surface is smooth. Aerial mycelium is light gray but substrate islight .olive brown. Produces brown diffusible pH sensitive pigments(brown ~ gray). Melanin negative. Growth IS sensitive to someinhibitors and antibiotics. Utilizes some carbon and nitrogen sources.Mesophile and neutrophile. Produces antimicrobial substances againstgram (+ve) bacteria and yeasts. Exhibits well lipolytic, keratinolytic,cellulolytic, pectinolytic and chitinolytic activities. Degrades someorganic complex compounds. It is identified as the new speciesStreptomyces einereobrunneus n.sp (Hussein and Mohammed 2005) .. Group (22): This group includes five isolates of which G-113 waschosen as the representative one. It produces spore chains of spores227---_._.-.consist of 2-5 turns, spores are of big size with abrupt ends and withsmooth surface. Aerial mycelium is light gray but substrate mycelium isbrownish gray. Brownish gray diffusible pigments are not pHsensitive. Melanin negative. Growth is sensitive to some inhibitors andantibiotics. Utilizes come carbon and nitrogen sources. Mesophile andneutrophile. Produces antimicrobial substances against gram (-ve), gram(+ve) bacteria, and yeasts. Exhibits well lipolytic, pectinolytic andchitinolytic activities. Degrades some organic complex compounds. It isidentifies as StreptQmyces pnoeocuromosenes (Krassilnikov, 1970).Group (23): This group contains only one isolate GnG6. Itproduces spiral chains of spores that have 4-6 turns. Spores are shortcylinders, with abrnpt ends, spore surface is provided with short spines.Aerial mycelium is light gray while substrate mycelium is brownish gray.The brownish gray diffusible pigments are not pH sensitive. Melaninnegative. Growth is sensitive to some inhibitors and antibiotics. IsolateGNG6 utilizes some carbon and nitrogen sources. Mesophile andneutrophile. Produces antimicrobial substances active against b>Tam(+ve)bacteria and yeasts. Exhibits well lipolytic, keratinolytic, pectinolytic andchitinolytic activities. Degrades some organic complex compounds. It isidentified as StreptQmyces varlabllis (Gauze et al., 1957).Blue section:Group (24): This b>TOUP contains 6 isolates of which G-14 wasconsidered as the representative one. It produces well developed spiralchains of spores spiral may be of 4-8 turns (multiwhorled), spores areshort moderate cylinders, spore surface ornamented with fine acutespines. Aerial mycelium is light greenish blue but substrate mycelium isgrayish green. No diffusible pigments. Melanin positive. Growth issensitive to some inhibitors and antibiotics. It utilizes some carbon and·’mti”dgenl’soUrces.Mesophile and neutrophile. It does not exhibit anyantimicrobial activities. Exhibits well lipolytic, keratinolytic, cellulolytic,pectinolytic and chitinolytic activities. Degrades some organic complexcompounds. It is identified as Streptomyces viridochromogenes(Krainsky, 1914).229