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Isolation, Identification And

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ﻣﺠﻠﻪ زﻳﺴﺖ ﺷﻨﺎﺳﻲ اﻳﺮان ﺟﻠﺪ 22، ﺷﻤﺎره 1، ﺑﻬﺎر 1388 ﺷﻨﺎﺳﺎﻳﻲ و ﺗﻌﻴﻴﻦ ﺧﺼﻮﺻﻴﺎت ﺑﺎﻛﺘﺮﻳﻬﺎي ﻧﻤﻚ دوﺳﺖ ﺗﻮﻟﻴﺪ ﻛﻨﻨﺪه آﻧﺰﻳﻤﻬﺎي ﻫﻴﺪروﻟﺘﻴﻚ ﺟﺪا ﺷﺪه از درﻳﺎﭼﻪ ﻧﻤﻚ آران و ﺑﻴﺪﮔﻞ 3 2 1 ﺣﻤﻴﺪ ﺑﺎﺑﺎوﻟﻴﺎن ، ﻣﺤﻤﺪ ﻋﻠﻲ آﻣﻮزﮔﺎر ،* و اﺣﻤﺪ ﻋﻠﻲ ﭘﻮرﺑﺎﺑﺎﺋﻲ 1 ﺗﻬﺮان، داﻧﺸﮕﺎه ﻋﻠﻮم ﭘﺰﺷﻜﻲ ﺑﻘﻴﻪ اﷲ (ﻋﺞ)، ﻣﺮﻛﺰ ﺗﺤﻘﻴﻘﺎت ﺑﻴﻮﺗﻜﻨﻮﻟﻮژي ﻛﺎرﺑﺮدي و ﻣﺤﻴﻂ زﻳﺴﺖ 2 ﺗﻬﺮان، داﻧﺸﮕﺎه ﺗﻬﺮان ، ﭘﺮدﻳﺲ ﻋﻠﻮم ، داﻧﺸﻜﺪه زﻳﺴﺖ ﺷﻨﺎﺳﻲ ، ﮔﺮوه ﻣﻴﻜﺮوﺑﻴﻮﻟﻮژي، آزﻣﺎﻳﺸﮕﺎه اﻛﺴﺘﺮﻣﻮﻓﻴﻞ 3 ﻗﻢ، داﻧﺸﮕﺎه آزاد اﺳﻼﻣﻲ، داﻧﺸﻜﺪه ﻋﻠﻮم ﮔﺮوه ﻣﻴﻜﺮوﺑﻴﻮﻟﻮژي ﺗﺎرﻳﺦ درﻳﺎﻓﺖ: 20/12/86 ﺗﺎرﻳﺦ ﭘﺬﻳﺮش: 87/9/6 ﭼﻜﻴﺪه درﻳﺎﭼﻪ آران و ﺑﻴﺪﮔﻞ ﻳﻜﻲ از درﻳﺎﭼﻪ ﻫﺎي ﺷﻮر در ﻣﻨﻄﻘﻪ ﻛﻮﻳﺮ ﻣﺮﻛﺰي اﻳﺮان ﻣﻲ ﺑﺎﺷﺪ. اﻳﻦ درﻳﺎﭼﻪ ﺷﻜﻠﻲ ﺷﺒﻴﻪ ﺑﻪ ﻳﻚ ﻣﺜﻠﺚ دارد ﻛﻪ رأس آن ﺑﻪ ﺳﻤﺖ ﺷﻤﺎل ﻣﻲ ﺑﺎﺷﺪ. ﻃﻮل ﻗﺎﻋﺪه اﻳﻦ ﻣﺜﻠﺚ 35 ﻛﻴﻠﻮﻣﺘﺮ و ارﺗﻔﺎع آن 38 ﻛﻴﻠﻮﻣﺘﺮ ﻣﻲ ﺑﺎﺷﺪ. ﻏﺮﺑﺎل ﮔﺮي ﺑﺎﻛﺘﺮﻳﻬﺎ از ﻧﻘﺎط ﻣﺨﺘﻠﻒ درﻳﺎﭼﻪ ﻣﻨﺠﺮ ﺑﻪ ﺟﺪاﺳﺎزي61 ﺑﺎﻛﺘﺮي ﮔﺮم ﻣﺜﺒﺖ و 22 ﺑﺎﻛﺘﺮي ﮔﺮم ﻣﻨﻔﻲ ﻧﻤﻚ دوﺳﺖ ﻧﺴﺒﻲ ﺗﻮاﻧﺎ ﺑﻪ ﺗﻮﻟﻴﺪ ﻫﻴﺪروﻻزﻫﺎي ﻣﺨﺘﻠﻒ ﺷﺪ اﻳﻦ ﺑﺎﻛﺘﺮﻳﻬﺎ ﺑﻪ ﻃﻮر اﭘﺘﻴﻤﻢ در ﻣﺤﻴﻂ ﺑﺎ 15- 5 درﺻﺪ ﻧﻤﻚ ، دﻣﺎي 37-34 درﺟﻪ ﺳﺎﻧﺘﻲ ﮔﺮاد و pH 2/7 رﺷﺪ ﻣﻲ ﻛﻨﻨﺪ ﺗﻌﺪاد ﺳﻮﻳﻪ ﻫﺎي ﺗﻮﻟﻴﺪ ﻛﻨﻨﺪه آﻧﺰﻳﻤﻬﺎي آﻣﻴﻼز، ﭘﺮوﺗﺌﺎز، ﻟﻴﭙﺎز، DNAase، اﻳﻨﻮﻟﻴﻨﺎز، ﮔﺰﻳﻼﻧﺎز، ﻛﺮﺑﻮﻛﺴﻲ ﻣﺘﻴﻞ ﺳﻠﻮﻻز، ﭘﻜﺘﻴﻨﺎز، ﭘﻮﻟﻮﻻﻧﺎز و ﻛﻴﺘﻴﻨﺎز ﺑﻪ ﺗﺮﺗﻴﺐ 32، 20، 27، 40، 40، 9، 11، 24، 16 و 20 ﺑﻮد ﻛﻪ در ﻣﻴﺎن اﻳﻦ ﺳﻮﻳﻪ ﻫﺎ ﺗﻌﺪادي ﻧﻴﺰﻗﺎدر ﺑﻪ ﺗﻮﻟﻴﺪ ﻣﺨﻠﻮﻃﻲ از اﻳﻦ آﻧﺰﻳﻤﻬﺎ ﺑﻮدﻧﺪ. ﺑﻴﺸﺘﺮﻳﻦ آﻧﺰﻳﻤﻬﺎي ﺗﻮﻟﻴﺪ ﺷﺪه در ﺑﺎﺳﻴﻠﻬﺎي ﮔﺮم ﻣﺜﺒﺖ آﻧﺰﻳﻤﻬﺎي DNase و اﻳﻨﻮﻟﻴﻨﺎز و در ﺑﺎﺳﻴﻠﻬﺎي ﮔﺮم ﻣﻨﻔﻲ آﻧﺰﻳﻢ ﻟﻴﭙﺎز و در ﻛﻮﻛﻮﺳﻬﺎي ﮔﺮم ﻣﺜﺒﺖ آﻧﺰﻳﻤﻬﺎي ﭘﻮﻟﻮﻻﻧﺎزو ﻛﺮﺑﻮﻛﺴﻲ ﻣﺘﻴﻞ ﺳﻠﻮﻻز ﺑﻮدﻧﺪ . ﻛﻤﺘﺮﻳﻦ ﻣﻴﺰان آﻧﺰﻳﻢ ﺗﻮﻟﻴﺪ ﺷﺪه ﻣﺮﺑﻮط ﺑﻪ آﻧﺰﻳﻢ ﮔﺰﻳﻼﻧﺎز ﺑﻮد . از اﻳﻦ ﺳﻮﻳﻪ ﻫﺎ، ﭼﻨﺪﻳﻦ ﺳﻮﻳﻪ ﺑﺎ ﺗﻮاﻧﺎﻳﻲ ﺗﻮﻟﻴﺪ ﺗﻨﻮﻋﻲ از آﻧﺰﻳﻤﻬﺎي ارزﺷﻤﻨﺪ اﻧﺘﺨﺎب ﺷﺪﻧﺪ ﻛﻪ ﺷﺎﻣﻞ AMB1 (8 آﻧﺰﻳﻢ) وAMB7, AMB10, AMB1(7 آﻧﺰﻳﻢ) و ,AMB5, AMB6, AMB11 AMB13 (6 آﻧﺰﻳﻢ) وAMB2, AMB3, AMB8, AMB9 (5 آﻧﺰﻳﻢ) و AMB4, AMG1 (4 آﻧﺰﻳﻢ) و AMC1, AMC2 (3 آﻧﺰﻳﻢ) و AMG2 ،AMC17 ،AMC8 ،AMC3 (2 آﻧﺰﻳﻢ) و AMG3 (1 آﻧﺰﻳﻢ) ﺑﻮدﻧﺪ. ﻧﻬﺎﻳﺘﺎً ﺳﻮﻳﻪ ﻫﺎي ﻣﻨﺘﺨﺐ ﻋﻼوه ﺑﺮ ﺻﻔﺎت ﻓﻨﻮﺗﻴﭙﻲ از ﻧﻈﺮ وﻳﮋﮔﻴﻬﺎي ﻓﻴﻠﻮژﻧﻲ و ﻣﻮﻟﻜﻮﻟﻲ ﺑﺎ ﺗﻜﻨﻴﻚ 16S rDNA ﺷﻨﺎﺳﺎﻳﻲ و ﺗﻌﻴﻴﻦ ﺗﻮاﻟﻲ ﺷﺪﻧﺪ، ﻛﻪ ﺑﺎﻛﺘﺮﻳﻬﺎي ﮔﺮم ﻣﺜﺒﺖ ﻣﺘﻌﻠﻖ ﺑﻪ ﺟﻨﺴﻬﺎي Nesterenkonia ، Halobacillus, Thalassobacillus, Bacillus, Salinicoccus، Staphylococcus ،Marinococcus و ﺑﺎﻛﺘﺮﻳﻬﺎي ﮔﺮم ﻣﻨﻔﻲ ﻣﺘﻌﻠﻖ ﺑﻪ ﺟﻨﺴﻬﺎي Idiomarina, Salicola و Halomonas ﺑﻮدﻧﺪ. ﺑﺎﻛﺘﺮي ﺟﻨﺲ Halobacillus ﻓﺮاوان ﺗﺮﻳﻦ ﺑﺎﻛﺘﺮي ﺟﺪا ﺷﺪه از درﻳﺎﭼﻪ و ﺑﻴﺸﺘﺮﻳﻦ ﺗﻨﻮع آﻧﺰﻳﻢ را ﻧﺸﺎن داد . واژه ﻫﺎي ﻛﻠﻴﺪي: اﻛﺴﺘﺮﻣﻮﻓﻴﻞ، ﻧﻤﻚ دوﺳﺖ، آﻧﺰﻳﻤﻬﺎي ﻫﻴﺪروﻟﻴﺘﻴﻚ، ﻏﺮﺑﺎل ﮔﺮي *ﻧﻮﻳﺴﻨﺪه ﻣﺴﺌﻮل، ﺗﻠﻔﻦ ﺗﻤﺎس 02161112723، ﭘﺴﺖ اﻟﻜﺘﺮوﻧﻴﻚ: [email protected] ﻣﻘﺪﻣﻪ درﻳﺎﭼﻪ آران و ﺑﻴﺪﮔﻞ ﻳﻜﻲ از درﻳﺎﭼﻪ ﻫﺎ در ﻣﻨﻄﻘﻪ ﻛﻮﻳﺮ ﺳﻤﺖ ﺷﻤﺎل ﻣﻲ ﺑﺎﺷﺪ . ﻃﻮل ﻗﺎﻋﺪه اﻳﻦ ﻣﺜﻠﺚ 35 ﻛﻴﻠﻮﻣﺘﺮ ﻣﺮﻛﺰي اﻳﺮان و ﺑﺰرﮔﺘﺮﻳﻦ ﭘﻼﻳﺎي ﺷﻮر در اﻳﺮان اﺳﺖ. اﻳﻦ و ارﺗﻔﺎع آن 38 ﻛﻴﻠﻮﻣﺘﺮ ﻣﻲ ﺑﺎﺷﺪ . ﻋﻤﻖ ﻧﻤﻚ آن ﺑﻴﻦ 54 - درﻳﺎﭼﻪ ﺷﻜﻠﻲ ﺷﺒﻴﻪ ﺑﻪ ﻳﻚ ﻣﺜﻠﺚ دارد، ﻛﻪ رأس آن ﺑﻪ 5 ﻣﺘﺮ ﻣﺘﻐﻴﻴﺮ ﻣﻲ ﺑﺎﺷﺪ ﻛﻪ ﺗﻮﺳﻂ ﻻﻳﻪ ﻫﺎي ﺧﺎك رس از 24 ﻣﺠﻠﻪ زﻳﺴﺖ ﺷﻨﺎﺳﻲ اﻳﺮان ﺟﻠﺪ 22، ﺷﻤﺎره 1، ﺑﻬﺎر 1388 ﻳﻜﺪﻳﮕﺮ ﺟﺪا ﺷﺪه اﻧﺪ. ﺗﺮﻛﻴﺒﺎت ﺗﺸﻜﻴﻞ دﻫﻨﺪه ﻧﻤﻚ اﻧﺪ(23)، در اﻳﻦ ﻣﻄﺎﻟﻌﻪ ﺗﻨﻮع زﻳﺴﺘﻲ ﺑﺎﻛﺘﺮﻳﻬﺎي ﻧﻤﻚ درﻳﺎﭼﻪ، ﻣﻮادي از ﻗﺒﻴﻞ ﺳﺪﻳﻢ ﻛﻠﺮاﻳﺪ، ﺳﺪﻳﻢ ﺳﻮﻟﻔﺎت، دوﺳﺖ ﺟﺪا ﺷﺪه از ﺑﺨﺸﻬﺎي ﻣﺨﺘﻠﻒ در درﻳﺎﭼﻪ ﻧﻤﻚ آران ﻣﻨﻴﺰﻳﻢ ﻛﻠﺮاﻳﺪ، ﻣﻨﻴﺰﻳﻢ ﺳﻮﻟﻔﺎت و .... ﺑﻮده و ﺑﻴﺸﺘﺮ ﻓﺼﻮل و ﺑﻴﺪﮔﻞ ﺗﻮﺻﻴﻒ ﺷﺪه اﺳﺖ. ﺳﺎل ﺧﺸﻚ و اﺷﺒﺎع از ﻧﻤﻚ اﺳﺖ. وﻳﮋﮔﻴﻬﺎي ﺷﺎﺧﺺ اﻳﻦ ﻣﻮاد و روﺷﻬﺎ درﻳﺎﭼﻪ از ﺟﻨﺒﻪ ﺷﻴﻤﻴﺎﻳﻲ، ﺑﺎﻻ ﺑﻮدن ﻋﻴﺎر ﻣﻨﻴﺰﻳﻢ آن ﻧﺴﺒﺖ ﺑﻪ ﺳﺎﻳﺮ درﻳﺎﭼﻪ ﻫﺎ و ﺷﻮراﺑﻪ ﻫﺎي ﻣﻄﺎﻟﻌﻪ ﺷﺪه در اﻳﺮان، از ﻧﻤﻮﻧﻪ ﮔﻴﺮي از ﻧﻘﺎط ﻣﺨﺘﻠﻒ درﻳﺎﭼﻪ: ﺑﻪ ﻣﻨﻈﻮر ﺟﺪاﺳﺎزي ﺟﻨﺒﻪ ﺑﻴﻮﻟﻮژﻳﻚ، ﻣﻴﺰان ﺷﻮري اﺷﺒﺎع ﻧﻤﻚ و ﻻﻳﻪ ﻫﺎي رﻧﮕﻲ ﺳﻮﻳﻪ ﻫﺎي ﻧﻤﻚ دوﺳﺖ ﺑﺎ ﺗﻮاﻧﺎﻳﻲ ﺗﻮﻟﻴﺪ آﻧﺰﻳﻤﻬﺎي ﻧﻤﻚ (ﻗﺮﻣﺰ، ﻧﺎرﻧﺠﻲ، ﺳﺒﺰ، ﺳﻴﺎه) و از ﺟﻨﺒﻪ اﻗﺘﺼﺎدي، ﻫﻴﺪروﻟﻴﺘﻴﻚ ﺧﺎرج ﺳﻠﻮﻟﻲ، از ﻧﻘﺎط ﻣﺨﺘﻠﻒ درﻳﺎﭼﻪ ﻧﻤﻚ ﺣﺼﻮل ﻣﻘﺪار زﻳﺎد ﻧﻤﻚ را ﻣﻲ ﺗﻮان ﻧﺎم ﺑﺮد. اﻳﻦ درﻳﺎﭼﻪ ﺑﻪ آران و ﺑﻴﺪﮔﻞ(از 5 ﻧﻘﻄﻪ درﻳﺎﭼﻪ ﻃﺒﻖ ﻋﻼﻣﺖ ﮔﺬاري روي ﻋﻠﺖ ﺑﺎﻻ ﺑﻮدن ﻣﻴﺰان ﺷﻮري ﻳﻚ زﻳﺴﺘﮕﺎه ﻣﻨﺎﺳﺐ ﺑﺮاي ﻧﻘﺸﻪ ﺑﺎ ﻧﺎﻣﻬﺎي ﻣﺴﻴﺮ ﻣﻨﺘﻬﻲ ﺑﻪ درﻳﺎﭼﻪ ، ﻣﻨﻄﻘﻪ ﺟﺰﻳﺮه دﺳﺘﻪ اي از اﻛﺴﺘﺮﻣﻮﻓﻴﻞ ﻫﺎ ﺑﻪ ﻧﺎم ﻧﻤﻚ دوﺳﺘﻬﺎ ﻣﻲ ﺑﺎﺷﺪ. ﺳﺮﮔﺮداﻧﻲ، ﻣﻨﻄﻘﻪ ﻧﻤﻚ ﭘﺎرﺳﻴﺎن، ﻣﻨﻄﻘﻪ ﻧﻤﻚ ﻛﺎﻧﺴﺎر و ﺑﺎﻛﺘﺮﻳﻬﺎي ﻧﻤﻚ دوﺳﺖ ﻣﻴﻜﺮوارﮔﺎﻧﻴﺴﻢ ﻫﺎﻳﻲ ﻫﺴﺘﻨﺪ ﻛﻪ ﻣﻲ ﻣﻨﻄﻘﻪ 18 ﻛﻴﻠﻮﻣﺘﺮي) در دو ﻣﺎه آﺑﺎن و اردﻳﺒﻬﺸﺖ ﻧﻤﻮﻧﻪ ﺗﻮاﻧﻨﺪ ﺑﺼﻮرت ﺑﻬﻴﻨﻪ در ﻣﺤﻴﻂ ﻣﺤﺘﻮي 15 -5 درﺻﺪ ﻧﻤﻚ ﮔﻴﺮي اﻧﺠﺎم ﺷﺪ. ﺷﻜﻞ 1 ﻧﻘﺎط ﻧﻤﻮﻧﻪ ﺑﺮداري را ﺑﺮ روي رﺷﺪ ﻛﻨﻨﺪ (8 و21). اﻟﺒﺘﻪ اﻳﻦ ﻣﺤﺪوده ﻧﻤﻚ ﺑﺴﺘﮕﻲ ﺑﻪ ﻧﻘﺸﻪ و در ﺟﺪول 1 ﻣﺸﺨﺼﺎت ﻧﻘﺎط ﻧﻤﻮﻧﻪ ﺑﺮداري ﻧﺸﺎن ﺗﺮﻛﻴﺒﺎت ﻣﺤﻴﻂ ﻛﺸﺖ و دﻣﺎ دارد. آﻧﻬﺎ ﺑﻪ ﻃﻮر وﺳﻴﻊ در داده ﺷﺪه اﺳﺖ. ﻓﺮاواﻧﻲ ﺳﻮﻳﻪ ﻫﺎي ﺟﺪا ﺷﺪه از ﻣﺤﻠﻬﺎي زﻳﺴﺘﮕﺎﻫﻬﺎي ﺷﻮر ﻣﺎﻧﻨﺪ درﻳﺎﭼﻪ ﻫﺎي ﺷﻮر، ﺧﺎﻛﻬﺎ و ﻧﻤﻮﻧﻪ ﺑﺮداري در ﺟﺪول 2 ﻧﺸﺎن داده ﺷﺪه اﺳﺖ . ﺑﻴﺎﺑﺎﻧﻬﺎي ﻧﻤﻜﺰار، ﺣﻮﺿﭽﻪ ﻫﺎي ﺑﺮداﺷﺖ ﻧﻤﻚ، ﻣﻌﺎدن ﺳﻮﻳﻪ ﻫﺎي ﺑﺎﻛﺘﺮﻳﺎﻳﻲ و ﺷﺮاﻳﻂ رﺷﺪ: ﺟﺪاﺳﺎزي ﺑﺎﻛﺘﺮﻳﻬﺎي 9 ﻧﻤﻚ، ﻏﺬاﻫﺎي ﺷﻮر و دﻳﮕﺮ ﻣﻨﺎﻃﻖ ﭘﺮاﻛﻨﺪه ﻫﺴﺘﻨﺪ ( ﻧﻤﻚ دوﺳﺖ ﺑﺮ روي ﻣﺤﻴﻂ ﻣﻐﺬي آﮔﺎر داري ﺑﻮد ﻛﻪ ﺑﺪان 22 و ). ﺟﻨﺒﻪ ﻫﺎﻳﻲ ﻛﻪ اﺧﻴﺮاً ﺗﻮﺟﻪ ﻣﺤﻘﻘﻴﻦ را ﺑﻪ ﺧﻮد ﺟﻠﺐ ﻧﻤﻜﻬﺎي زﻳﺮ اﻓﺰوده ﺷﺪه اﺳﺖ . ﻛﺮده اﺳﺖ ﺑﻴﺸﺘﺮ در ارﺗﺒﺎط ﺑﺎ ﻣﻜﺎﻧﻴﺴﻢ ﻓﻴﺰﻳﻮﻟﻮژي آداﭘﺘﻪ NaCl,81 g/l ; 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درﺻﺪStarch agar, 30 g/l ; NaCl (solar salt), 10 ﭘﻮﺷﻴﺪه ﺷﺪ، ﻇﻬﻮر ﻳﻚ ﻧﺎﺣﻴﻪ ﺷﻔﺎف اﻃﺮاف ﻛﻠﻨﻲ ﻫﺎ ﻧﺸﺎﻧﻪ Distilled water, 1000ml ;pH, 7.2 ﻫﻴﺪروﻟﻴﺰ ﻧﺸﺎﺳﺘﻪ ﻣﻲ ﺑﺎﺷﺪ (5 ). ). ﺑﻌﺪ از اﻧﻜﻮﺑﺎﺳﻴﻮن در دﻣﺎي 37 درﺟﻪ ﺳﺎﻧﺘﻲ ﮔﺮاد ﺑﻪ ﻣﺪت 7 روز، ﭘﻠﻴﺖ ﻫﺎ ﺑﺎ ﻣﺤﻠﻮل (0.3) درﺻﺪ I2, 0.6 درﺻﺪ KI 27 ﻣﺠﻠﻪ زﻳﺴﺖ ﺷﻨﺎﺳﻲ اﻳﺮان ﺟﻠﺪ 22، ﺷﻤﺎره 1، ﺑﻬﺎر 1388 ﺗﻮﻟﻴﺪ ﺧﺎرج ﺳﻠﻮﻟﻲ ﭘﺮوﺗﺌﺎز: Skim milk, 10 درﺻﺪ ; رﺷﺪ در ﺳﻄﺢ ﭘﻠﻴﺖ ﻧﺸﺎﻧﻪ ﺗﻮﻟﻴﺪ آﻧﺰﻳﻢ و اﺳﺘﻔﺎده از اﻳﻨﻮﻟﻴﻦ N.A , 28 g/l ; NaCl (solar salt), 10 درﺻﺪ Agar ,15 ; ﺑﻪ ﻋﻨﻮان ﺗﻨﻬﺎ ﻣﻨﺒﻊ ﻛﺮﺑﻦ ﻣﻲ ﺑﺎﺷﺪ. (1 ) ) g/l ; Distilled water, 1000ml ; pH 7.2 ﺗﻮﻟﻴﺪ ﺧﺎرج ﺳﻠﻮﻟﻲ ﭘﻜﺘﻴﻨﺎز: ﻣﺤﻴﻂ ﻛﺸﺖ ﺗﻮﻟﻴﺪ آﻧﺰﻳﻢ ﺗﺮﻛﻴﺒﺎت ﻣﺤﻴﻂ ﺑﻪ ﺟﺰ Skim milk در 121 درﺟﻪ ﺳﺎﻧﺘﻲ ﭘﻜﺘﻴﻨﺎز ﮔﺮاد 15 دﻗﻴﻘﻪ اﺗﻮﻛﻼو ﮔﺸﺖ و skim milk از ﻃﺮﻳﻖ Pectin, 1 ; (NH ) SO , 0.14 ;K HPO , ﺻﺎف ﻛﺮدن اﺳﺘﺮﻳﻞ ﮔﺸﺖ . 4 2 درﺻﺪ 4 2 4 درﺻﺪ Nutrient ; درﺻﺪMgSO4.7H2O, 0.02 ; درﺻﺪ 0.2 اﻳﺠﺎد ﻫﺎﻟﻪ ﺷﻔﺎف اﻃﺮاف ﻛﻠﻨﻲ ﻫﺎ ﻧﺸﺎﻧﻪ ﺗﻮﻟﻴﺪ آﻧﺰﻳﻢ ﻣﻲ ; FeSO4.7H2O, 5 mg/l ) درﺻﺪ solution, 0.1 20 ﺑﺎﺷﺪ. ( ) ; MnSO4.H2O, 1.6 mg/L ; ZnSO4.7H2O, 1.4 mg/l ,Agar ; درﺻﺪ CaCL2, 2 mg/l ) ;NaCl(solar salt), 10 ﺗﻮﻟﻴﺪ ﺧﺎرج ﺳﻠﻮﻟﻲ ﻟﻴﭙﺎز: .Peptone , 10 g/l ; CaCl2 2 ; Distilled water, 1000ml; pH 7. 2 ,(H2O, 0.1 g/l ; Tween 80, 10 g/l ; NaCl (solar salt درﺻﺪ 10 درﺻﺪDistilled water, 1000ml; pH, 7.2 ; ﭘﻜﺘﻴﻦ از ﻃﺮﻳﻖ ﺻﺎف ﻛﺮدن اﺳﺘﺮﻳﻞ و ﺑﻘﻴﻪ ﻣﺤﻴﻂ در دﻣﺎي ﺑﻌﺪ از ﺗﻬﻴﻪ ﻣﺤﻴﻂ ﻓﻮق در دﻣﺎي 115 درﺟﻪ ﺳﺎﻧﺘﻲ ﮔﺮاد ﺑﻪ 121 درﺟﻪ ﺑﻪ ﻣﺪت 15 دﻗﻴﻘﻪ اﺳﺘﺮﻳﻞ ﺷﺪ .
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  • Diversity of Culturable Moderately Halophilic and Halotolerant Bacteria in a Marsh and Two Salterns a Protected Ecosystem of Lower Loukkos (Morocco)

    Diversity of Culturable Moderately Halophilic and Halotolerant Bacteria in a Marsh and Two Salterns a Protected Ecosystem of Lower Loukkos (Morocco)

    African Journal of Microbiology Research Vol. 6(10), pp. 2419-2434, 16 March, 2012 Available online at http://www.academicjournals.org/AJMR DOI: 10.5897/ AJMR-11-1490 ISSN 1996-0808 ©2012 Academic Journals Full Length Research Paper Diversity of culturable moderately halophilic and halotolerant bacteria in a marsh and two salterns a protected ecosystem of Lower Loukkos (Morocco) Imane Berrada1,4, Anne Willems3, Paul De Vos3,5, ElMostafa El fahime6, Jean Swings5, Najib Bendaou4, Marouane Melloul6 and Mohamed Amar1,2* 1Laboratoire de Microbiologie et Biologie Moléculaire, Centre National pour la Recherche Scientifique et Technique- CNRST, Rabat, Morocco. 2Moroccan Coordinated Collections of Micro-organisms/Laboratory of Microbiology and Molecular Biology, Rabat, Morocco. 3Laboratory of Microbiology, Faculty of Sciences, Ghent University, Ghent, Belgium. 4Faculté des sciences – Université Mohammed V Agdal, Rabat, Morocco. 5Belgian Coordinated Collections of Micro-organisms/Laboratory of Microbiology of Ghent (BCCM/LMG) Bacteria Collection, Ghent University, Ghent, Belgium. 6Functional Genomic plateform - Unités d'Appui Technique à la Recherche Scientifique, Centre National pour la Recherche Scientifique et Technique- CNRST, Rabat, Morocco. Accepted 29 December, 2011 To study the biodiversity of halophilic bacteria in a protected wetland located in Loukkos (Northwest, Morocco), a total of 124 strains were recovered from sediment samples from a marsh and salterns. 120 isolates (98%) were found to be moderately halophilic bacteria; growing in salt ranges of 0.5 to 20%. Of 124 isolates, 102 were Gram-positive while 22 were Gram negative. All isolates were identified based on 16S rRNA gene phylogenetic analysis and characterized phenotypically and by screening for extracellular hydrolytic enzymes. The Gram-positive isolates were dominated by the genus Bacillus (89%) and the others were assigned to Jeotgalibacillus, Planococcus, Staphylococcus and Thalassobacillus.
  • Exploring Bacterial Community Composition in Mediterranean Deep-Sea Sediments and Their Role in Heavy Metal Accumulation

    Exploring Bacterial Community Composition in Mediterranean Deep-Sea Sediments and Their Role in Heavy Metal Accumulation

    Science of the Total Environment 712 (2020) 135660 Contents lists available at ScienceDirect Science of the Total Environment journal homepage: www.elsevier.com/locate/scitotenv Exploring bacterial community composition in Mediterranean deep-sea sediments and their role in heavy metal accumulation Fadwa Jroundi a,⁎, Francisca Martinez-Ruiz b,MohamedL.Merrouna, María Teresa Gonzalez-Muñoz a a Department of Microbiology, Faculty of Science, University of Granada, Avda. Fuentenueva s/n, 18071 Granada, Spain b Instituto Andaluz de Ciencias de la Tierra (CSIC-UGR), Av. de las Palmeras 4, 18100 (Armilla) Granada, Spain HIGHLIGHTS GRAPHICAL ABSTRACT • The westernmost Mediterranean is highly sensitive to anthropogenic pres- sure. • NGS showed mostly Bacillus and Micro- coccus as dominant in the deep-sea sed- iments. • Culturable bacteria revealed mostly the presence of Firmicutes and Actinobacteria. • Marine culturable bacteria bioaccumulate heavy metals within the cells and/or in EPS. • Lead precipitates in the sediment bacte- rial cells as pyromorphite. article info abstract Article history: The role of microbial processes in bioaccumulation of major and trace elements has been broadly demonstrated. Received 25 September 2019 However, microbial communities from marine sediments have been poorly investigated to this regard. In marine Received in revised form 18 November 2019 environments, particularly under high anthropogenic pressure, heavy metal accumulation increases constantly, Accepted 19 November 2019 which may lead to significant environmental issues. A better knowledge of bacterial diversity and its capability to Available online 22 November 2019 bioaccumulate metals is essential to face environmental quality assessment. The oligotrophic westernmost Med- Editor: Dr. Frederic Coulon iterranean, which is highly sensitive to environmental changes and subjected to increasing anthropogenic pres- sure, was selected for this study.
  • Bioprospecting for Novel Halophilic and Halotolerant Sources of Hydrolytic Enzymes in Brackish, Saline and Hypersaline Lakes of Romania

    Bioprospecting for Novel Halophilic and Halotolerant Sources of Hydrolytic Enzymes in Brackish, Saline and Hypersaline Lakes of Romania

    microorganisms Article Bioprospecting for Novel Halophilic and Halotolerant Sources of Hydrolytic Enzymes in Brackish, Saline and Hypersaline Lakes of Romania Robert Ruginescu 1,*, Ioana Gomoiu 1, Octavian Popescu 1,2, Roxana Cojoc 1, Simona Neagu 1, Ioana Lucaci 1, Costin Batrinescu-Moteau 1 and Madalin Enache 1 1 Department of Microbiology, Institute of Biology Bucharest of the Romanian Academy, 296 Splaiul Independentei, P.O. Box 56-53, 060031 Bucharest, Romania; [email protected] (I.G.); [email protected] (O.P.); [email protected] (R.C.); [email protected] (S.N.); [email protected] (I.L.); [email protected] (C.B.-M.); [email protected] (M.E.) 2 Molecular Biology Center, Institute of Interdisciplinary Research in Bio-Nano-Sciences, Babes-Bolyai-University, 42 Treboniu Laurian St., 400271 Cluj-Napoca, Romania * Correspondence: [email protected] Received: 4 November 2020; Accepted: 30 November 2020; Published: 30 November 2020 Abstract: Halophilic and halotolerant microorganisms represent promising sources of salt-tolerant enzymes that could be used in various biotechnological processes where high salt concentrations would otherwise inhibit enzymatic transformations. Considering the current need for more efficient biocatalysts, the present study aimed to explore the microbial diversity of five under- or uninvestigated salty lakes in Romania for novel sources of hydrolytic enzymes. Bacteria, archaea and fungi were obtained by culture-based approaches and screened for the production of six hydrolases (protease, lipase, amylase, cellulase, xylanase and pectinase) using agar plate-based assays. Moreover, the phylogeny of bacterial and archaeal isolates was studied through molecular methods. From a total of 244 microbial isolates, 182 (74.6%) were represented by bacteria, 22 (9%) by archaea, and 40 (16.4%) by fungi.
  • Thermolongibacillus Cihan Et Al

    Thermolongibacillus Cihan Et Al

    Genus Firmicutes/Bacilli/Bacillales/Bacillaceae/ Thermolongibacillus Cihan et al. (2014)VP .......................................................................................................................................................................................... Arzu Coleri Cihan, Department of Biology, Faculty of Science, Ankara University, Ankara, Turkey Kivanc Bilecen and Cumhur Cokmus, Department of Molecular Biology & Genetics, Faculty of Agriculture & Natural Sciences, Konya Food & Agriculture University, Konya, Turkey Ther.mo.lon.gi.ba.cil’lus. Gr. adj. thermos hot; L. adj. Type species: Thermolongibacillus altinsuensis E265T, longus long; L. dim. n. bacillus small rod; N.L. masc. n. DSM 24979T, NCIMB 14850T Cihan et al. (2014)VP. .................................................................................. Thermolongibacillus long thermophilic rod. Thermolongibacillus is a genus in the phylum Fir- Gram-positive, motile rods, occurring singly, in pairs, or micutes,classBacilli, order Bacillales, and the family in long straight or slightly curved chains. Moderate alka- Bacillaceae. There are two species in the genus Thermo- lophile, growing in a pH range of 5.0–11.0; thermophile, longibacillus, T. altinsuensis and T. kozakliensis, isolated growing in a temperature range of 40–70∘C; halophile, from sediment and soil samples in different ther- tolerating up to 5.0% (w/v) NaCl. Catalase-weakly positive, mal hot springs, respectively. Members of this genus chemoorganotroph, grow aerobically, but not under anaer- are thermophilic (40–70∘C), halophilic (0–5.0% obic conditions. Young cells are 0.6–1.1 μm in width and NaCl), alkalophilic (pH 5.0–11.0), endospore form- 3.0–8.0 μm in length; cells in stationary and death phases ing, Gram-positive, aerobic, motile, straight rods. are 0.6–1.2 μm in width and 9.0–35.0 μm in length.
  • Isolation of Cultivable Halophilic Bacillus Sp. from the Makgadikgadi Salt Pans in Botswana

    Isolation of Cultivable Halophilic Bacillus Sp. from the Makgadikgadi Salt Pans in Botswana

    ® The African Journal of Plant Science and Biotechnology ©2011 Global Science Books Isolation of Cultivable Halophilic Bacillus sp. from the Makgadikgadi Salt Pans in Botswana Bakang Baloi • Maitshwarelo Ignatius Matsheka* • Berhanu Abegaz Gashe Department of Biological Sciences, University of Botswana, Pr Bag UB00704, Gaborone, Botswana Corresponding author : * [email protected] ABSTRACT Halophilic bacteria from the Makgadikgadi salt pans in north central Botswana were isolated using culture-dependent methods. Polymerase chain reaction (PCR) amplification of the 16S rRNA gene and phylogenetic analysis were used to identify the strains. Culturing was done aerobically in six different complex salt media. Salt concentrations used were 15, 20, 25 and 30% (2.6, 3.4, 4.3 and 5.1 M, respectively) NaCl, at pH 7.2 to pH 8.0. Four colony morphology types were isolated in axenic cultures comprising Gram-positive cells. Universal bacterial primers were used to amplify 16S rDNA from chromosomal DNA isolated from three of the four distinct colony groups. Restriction enzyme digests analysis of the 16S rDNA revealed seven RFLP types. Five of the RFLP types were subjected to sequencing. Comparison of the 16S rDNA sequence alignment to reference sequence data bases showed samples S2012A3, S2012B2 and S2012B3 to have between 95 and 99% homology to Bacillus sp. BH 164 and Bacillus sp. HS 136T, a novel species recently described as Bacillus persepolensis. Isolate S4102D4 showed 95 to 99% homology to Thalassobacillus sp. JY0201 and Thalassobacillus sp. FIB228 and Halobacillus sp. MO56 species. All five isolates had at least 95% similarity to published sequences implying they could be species within the described genera.
  • Isolation and Diversity of Sediment Bacteria in The

    Isolation and Diversity of Sediment Bacteria in The

    bioRxiv preprint doi: https://doi.org/10.1101/638304; this version posted May 14, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Isolation and Diversity of Sediment Bacteria in the 2 Hypersaline Aiding Lake, China 3 4 Tong-Wei Guan, Yi-Jin Lin, Meng-Ying Ou, Ke-Bao Chen 5 6 7 Institute of Microbiology, Xihua University, Chengdu 610039, P. R. China. 8 9 Author for correspondence: 10 Tong-Wei Guan 11 Tel/Fax: +86 028 87720552 12 E-mail: [email protected] 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 bioRxiv preprint doi: https://doi.org/10.1101/638304; this version posted May 14, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 29 Abstract A total of 343 bacteria from sediment samples of Aiding Lake, China, were isolated using 30 nine different media with 5% or 15% (w/v) NaCl. The number of species and genera of bacteria recovered 31 from the different media significantly varied, indicating the need to optimize the isolation conditions. 32 The results showed an unexpected level of bacterial diversity, with four phyla (Firmicutes, 33 Actinobacteria, Proteobacteria, and Rhodothermaeota), fourteen orders (Actinopolysporales, 34 Alteromonadales, Bacillales, Balneolales, Chromatiales, Glycomycetales, Jiangellales, Micrococcales, 35 Micromonosporales, Oceanospirillales, Pseudonocardiales, Rhizobiales, Streptomycetales, and 36 Streptosporangiales), including 17 families, 41 genera, and 71 species.
  • The Occurrence of Potential and Novel Isolates of Oceanobacillus Sp

    The Occurrence of Potential and Novel Isolates of Oceanobacillus Sp

    See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/343877969 The Occurrence of potential and novel isolates of Oceanobacillus sp. JAS12 and Salinicoccus sp. JS20 recovered from West Coast of Arabian Sea, India Article in Research Journal of Biotechnology · September 2020 CITATIONS READS 0 65 3 authors, including: Jayachandra Yaradoddi Agsar Dayanand KLE Technological University, Hubli Gulbarga University 48 PUBLICATIONS 130 CITATIONS 21 PUBLICATIONS 83 CITATIONS SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: Bacterial diversity and Biotechnological applications View project Bionanotechnology View project All content following this page was uploaded by Jayachandra Yaradoddi on 26 August 2020. The user has requested enhancement of the downloaded file. Research Journal of Biotechnology Vol. 15 (9) September (2020) Res. J. Biotech The Occurrence of potential and novel isolates of Oceanobacillus sp. JAS12 and Salinicoccus sp. JS20 recovered from West Coast of Arabian Sea, India Yaradoddi Jayachandra S.1,2,3*, Sulochana M.B.2, Kontro Merja H.3, Parameshwar A.B.2 and Agsar Dayanand4 1. Biomaterials Laboratory, Center for Materials Science, KLE Technological University, Hubballi-580031, INDIA 2. Department of Studies and Research in Biotechnology, Gulbarga University, Kalaburagi, Karnataka, 585106, INDIA 3. Ecology and Environmental Research Programme, University of Helsinki, Lahti, FINLAND 4. Department of Studies and Research in Microbiology, Gulbarga University, Kalaburagi, Karnataka, 585106, INDIA *[email protected] Abstract halophilic bacteria are the microbes that can grow under Many halophiles were considered to be extremophiles higher salt concentrations. Halophiles inherently possess due to their inborn industrial potentials and tolerance different industrially important metabolites producing to hostile environmental conditions.
  • Reorganising the Order Bacillales Through Phylogenomics

    Reorganising the Order Bacillales Through Phylogenomics

    Systematic and Applied Microbiology 42 (2019) 178–189 Contents lists available at ScienceDirect Systematic and Applied Microbiology jou rnal homepage: http://www.elsevier.com/locate/syapm Reorganising the order Bacillales through phylogenomics a,∗ b c Pieter De Maayer , Habibu Aliyu , Don A. Cowan a School of Molecular & Cell Biology, Faculty of Science, University of the Witwatersrand, South Africa b Technical Biology, Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology, Germany c Centre for Microbial Ecology and Genomics, University of Pretoria, South Africa a r t i c l e i n f o a b s t r a c t Article history: Bacterial classification at higher taxonomic ranks such as the order and family levels is currently reliant Received 7 August 2018 on phylogenetic analysis of 16S rRNA and the presence of shared phenotypic characteristics. However, Received in revised form these may not be reflective of the true genotypic and phenotypic relationships of taxa. This is evident in 21 September 2018 the order Bacillales, members of which are defined as aerobic, spore-forming and rod-shaped bacteria. Accepted 18 October 2018 However, some taxa are anaerobic, asporogenic and coccoid. 16S rRNA gene phylogeny is also unable to elucidate the taxonomic positions of several families incertae sedis within this order. Whole genome- Keywords: based phylogenetic approaches may provide a more accurate means to resolve higher taxonomic levels. A Bacillales Lactobacillales suite of phylogenomic approaches were applied to re-evaluate the taxonomy of 80 representative taxa of Bacillaceae eight families (and six family incertae sedis taxa) within the order Bacillales.