‫اﻟﺠﻤﻬﻮرﻳﺔ اﻟﻌﺮﺑﻴﺔ اﻟﺴﻮرﻳﺔ‬

‫هﻴﺌﺔ اﻟﻄﺎﻗـﺔ اﻟﺬرﻳـﺔ‬

‫هـ ط ذ س‪ -‬ب ج ‪ /‬ت ن ب ع ‪416‬‬

‫ﺗﺸﺮﻳﻦ اﻟﺜﺎﻧﻲ ‪2008‬‬

‫ﺗﻘﺮﻳﺮ ﻧﻬﺎﺋﻲ ﻋﻦ ﺑﺤﺚ ﻋﻠﻤﻲ‬

‫ﻗﺴﻢ اﻟﺒﻴﻮﻟﻮﺟﻴﺎ اﻟﺠﺰﺋﻴﺔ و اﻟﺘﻘﺎﻧﺔ اﻟﺤﻴﻮﻳﺔ‬

‫ﺗﻨﻤﻴﻂ ﺑﻌﺾ ﺳﻼﻻت ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﻓﻲ ﺳﻮرﻳﺔ ﺑﺎﺳﺘﺨﺪام ﺗﻘﺎﻧﺘﻲ‬

‫اﻟـ ‪ PCR‬واﻟـ ‪FT-IR‬‬

‫اﻟﺪآﺘﻮر أﻳﻤﻦ اﻟﻤﺮﻳﺮي‬

‫اﻷﺳﺘﺎذ اﻟﺪآﺘﻮر ﻧﺠﻢ اﻟﺪﻳﻦ ﺷﺮاﺑﻲ‬

‫هـ ط ذ س‪ -‬ب ج ‪ /‬ت ن ب ع ‪416‬‬

 ‫اﻟﻤﺸﺎرآﻮن ﻓﻲ اﻟﺪراﺳﺔ‬
‫ﺳﺎهﻢ ﻓﻲ إﻧﺠﺎز هﺬا اﻟﻌﻤﻞ آﻞ ﻣﻦ‪ :‬د‪ .‬ﻋﻬﺪ أﺑﻮ ﻳﻮﻧﺲ‪ ،‬واﻟﺴﻴﺪة رﻧﺪ ﻋﻜﻞ‪ ،‬واﻟﺴﻴﺪ‬
‫ﻓﻮاز اﻟﺸﻮﻟﻲ‪ ،‬واﻟﺴﻴﺪ ﻣﺤﻤﺪ ﺳﻌﻴﺪ‪ ،‬واﻟﺴﻴﺪ ﻣﺤﻤﺪ ﺻﺒﺮة‪ ،‬واﻟﺴﻴﺪ أﻧﺲ اﻟﻠﺤﺎم‪ ،‬ﻣﻦ‬
‫ﺧﻼل اﻟﻤﺸﺎرآﺔ ﻓﻲ اﻷﻋﻤﺎل اﻟﻤﺨﺒﺮﻳﺔ‪.‬‬

‫‪2‬‬
 ‫اﻟﻔﻬﺮس‬
‫اﻟﻤﺼﻄﻠﺤﺎت ‪4 ...............................................................................................................................‬‬
‫ﻣﺴﺘﺨﻠﺺ ‪5 ..................................................................................................................................‬‬
‫‪6 ...............................................................................................................................Summary‬‬
‫‪ .1‬اﻟﻤﻘﺪﻣﺔ ‪7 .................................................................................................................................‬‬
‫‪ 1.1‬ﺗﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ‪7 ..........................................................................‬‬
‫اﻟﻄﺮاﺋﻖ اﻟﺘﻘﻠﻴﺪﻳﺔ ﻟﺘﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ‪8 .......................................................‬‬
‫اﻟﻄﺮاﺋﻖ اﻟﺤﺪﻳﺜﺔ ﻟﺘﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ‪10 ......................................................‬‬
‫أﺟﻨﺎس وأﻧﻮاع ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ‪11 ......................................................................‬‬
‫‪ 4.1‬ﺑﻴﺌﺔ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ‪12 .............................................................................‬‬
‫‪ 4.2‬ﺗﺼﻨﻴﻊ اﻟﻠﺒﻦ ‪13 ............................................................................................‬‬
‫‪16‬‬ ‫‪ .2‬اﻟﻄﺮاﺋﻖ‬
‫‪ 1.2‬اﻻﻋﺘﻴﺎن ‪16 ................................................................................................‬‬
‫‪ 2.2‬اﻟﻌﺰل اﻟﺒﻜﺘﻴﺮي‪16 ........................................................................................‬‬
‫‪ 3.2‬اﻟﻔﺤﺺ اﻟﻤﺠﻬﺮي ‪16 .....................................................................................‬‬
‫‪ 4.2‬اﻻﺧﺘﺒﺎرات اﻟﻜﻴﻤﻴﺎﺋﻴﺔ اﻟﺤﻴﻮﻳﺔ ‪17 .......................................................................‬‬
‫‪ 5.2‬اﻻﺧﺘﺒﺎرات اﻟﺒﻴﻮﻟﻮﺟﻴﺔ اﻟﺠﺰﻳﺌﻴﺔ ‪17 ....................................................................‬‬
‫‪ .1.5.2‬ﻋﺰل اﻟـ ‪17 ..................................................................................DNA‬‬
‫‪ .2.5.2‬ﺗﻘﻨﻴﺔ اﻟـ ‪18 ................................................................................... PCR‬‬
‫‪ 6.2‬ﺗﻘﻨﻴﺔ اﻟـ ‪19 ........................................................................................ FT-IR‬‬
‫‪21‬‬ ‫‪ .3‬اﻟﻨﺘﺎﺋﺞ‬
‫‪ 1.3‬ﻧﺘﺎﺋﺞ ﺗﺤﺪﻳﺪ هﻮﻳﺔ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﺑﺘﻘﻨﻴﺔ اﻟـ ‪21 ........................................... PCR‬‬
‫‪ 2.3‬ﻧﺘﺎﺋﺞ ﻣﻄﻴﺎﻓﻴﺔ ﺗﺤﻮﻳﻞ ﻓﻮرﻳﻴﻪ ﻟﻸﺷﻌﺔ ﺗﺤﺖ اﻟﺤﻤﺮاء )‪24 ................................. (FT-IR‬‬
‫‪ .4‬اﻟﻤﻨﺎﻗﺸﺔ ‪36 ............................................................................................................................‬‬
‫آﻠﻤﺔ ﺷﻜﺮ ‪39 ................................................................................................................................‬‬
‫‪ .6‬اﻟﻤﺮاﺟﻊ ‪40 ..............................................................................................................................‬‬

‫‪3‬‬
 ‫اﻟﻤﺼﻄﻠﺤﺎت‬
‫‪Acetoin‬‬ ‫أﺳﻴﺘﻮﻳﻦ‬
‫‪Bacteria‬‬ ‫ﺑﻜﺘﻴﺮﻳﺎ‬
‫‪Cocci‬‬ ‫آﺮوي‬
‫‪Cyanobacteria‬‬ ‫اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﺰرﻗﺎء‬
‫‪FT-IR‬‬ ‫ﺗﺤﻮﻳﻞ ﻓﻮرﻳﻴﻪ ﻟﻸﺷﻌﺔ ﺗﺤﺖ اﻟﺤﻤﺮاء‬
‫‪Lactic Acid Bacteria‬‬ ‫ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‬
‫‪Milk‬‬ ‫ﺣﻠﻴﺐ‬
‫‪Rods‬‬ ‫ﻋﺼﻮي‬
‫‪Starter‬‬ ‫ﺑﺎدىء‬
‫‪Strain‬‬ ‫ﺳﻼﻟﺔ‬
‫‪Genus‬‬ ‫ﺟﻨﺲ‬
‫‪Species‬‬ ‫ﻧﻮع‬

‫‪4‬‬
 ‫ﻣﺴﺘﺨﻠﺺ‬
‫ﺗﻌﺘﺒ ﺮ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ﻣ ﻦ أآﺜ ﺮ اﻟﻤﺘﻌ ﻀﻴﺎت اﻟﻤﺠﻬﺮﻳ ﺔ ﻓﺎﺋ ﺪ ًة ﻟﻺﻧ ﺴﺎن‪ ،‬ﺣﻴ ﺚ ﻳﻤﻜ ﻦ‬
‫اﻻﺳﺘﻔﺎدة ﻣﻨﻬ ﺎ ﻓ ﻲ ﺗﺤ ﺴﻴﻦ ﻧﻜﻬ ﺔ ﺑﻌ ﺾ اﻷﻃﻌﻤ ﺔ‪ ،‬وﻓ ﻲ ﺗﺜﺒ ﻴﻂ ﺑﻌ ﺾ اﻟﻌﻮاﻣ ﻞ اﻟﻤﻤﺮﺿ ﺔ ورﺑﻤ ﺎ‬
‫ﻗﺘﻠﻬ ﺎ ﻓ ﻲ ﺑﻌ ﺾ ه ﺬﻩ اﻟﻤﻨﺘﺠ ﺎت‪ .‬ﻋُﺰﻟ ﺖ ﺳ ﻼﻻت ﻣ ﻦ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ﻣ ﻦ ﺑﻌ ﺾ ﻣﻨﺘﺠ ﺎت‬
‫اﻟﺤﻠﻴﺐ اﻟﺘﻘﻠﻴﺪﻳﺔ ﻓﻲ ﺳﻮرﻳﺔ‪ ،‬واﻟﺘﻲ ﺟُﻤﻌﺖ ﻋﻴﻨﺎﺗﻬﺎ ﻣﻦ ﻣﻨﺎﻃﻖ ﻣﺨﺘﻠﻔﺔ ﻣ ﻦ ﺳ ﻮرﻳﺔ‪ُ .‬د ِرس اﻟﻄ ﺎﺑﻊ‬
‫اﻟﻈﺎهﺮي ﻟﻬﺬﻩ اﻟﺴﻼﻻت واﺳْ ُﺘﺨﺪﻣﺖ ﺗﻘﻨﻴ ﺔ اﻟﺘﻔﺎﻋ ﻞ اﻟﺴﻠ ﺴﻠﻲ ﻟﻠﺒ ﻮﻟﻴﻤﻴﺮاز ‪ PCR‬ﻟﺘﻨﻤﻴﻄﻬ ﺎ‪ ،‬ﺣﻴ ﺚ‬
‫ﺟ َﺪت اﻷﻧﻮاع ‪faecium, E. faecalis‬‬
‫اﺧﺘﻴﺮت ﻣﻮرﺛﺔ ﻣﺤﺎﻓﻈﺔ ﻟﻠﻨﻮع ﻟﺪى ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪ُ .‬و ِ‬
‫‪ E.‬و‪ S. thermophilus‬ﻓﻲ اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ وﻓﻲ اﻟﻠﺒﻦ اﻟﺮاﺋ ﺐ‪ .‬وﻗ ﺪ أوﺿ ﺤﺖ ﻧﺘﺎﺋﺠﻨ ﺎ أﻧ ﻪ ﻳﻤﻜﻨﻨ ﺎ‬
‫اﺳﺘﺨﺪام ﻣﻄﻴﺎﻓﻴﺔ ﺗﺤﻮﻳﻞ ﻓﻮرﻳﻴﻪ ﻟﻸﺷ ﻌﺔ ﺗﺤ ﺖ اﻟﺤﻤ ﺮاء )‪ (FT-IR‬ﻟﺘﺤﺪﻳ ﺪ أﻧ ﻮاع ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ‬
‫اﻟﻠﺒﻦ‪.‬‬

‫آﻠﻤﺎت ﻣﻔﺘﺎح‪:‬‬
‫ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪ ،‬ﺗﻘﻨﻴﺔ اﻟـ ‪ ،API‬ﺗﻔﺎﻋﻞ اﻟﺘﻀﺨﻴﻢ اﻟﻤﻮرﺛﻲ‪ ،‬اﻟـ ‪.FT-IR‬‬

‫‪5‬‬
 Typing some of lactic acid bacteria in Syria using PCR and FT-IR
techniques

Summary
Lactic Acid Bacteria (LAB) are considered to be the most useful
microorganisms. They are beneficial in flavoring foods, inhibiting
pathogenic as well as spoilage bacteria in food products. The isolates of
LAB were obtained from traditional Syrain dairy products (white cheese
and curdled yogurt) obtained from different regions in Syria. The isolates
were subjected to phenotypic characterization analyses. The PCR technique
of bacterial DNA was evaluated as an advanced tool for the identification of
LAB. It was found that strains: E. faecium, E. faeccalis and S. thermophilus
dominate in white cheese and in yogurt. Our resultes demonstred that we
could identify LAB using Fourier transform infrared spectroscopy (FT-IR)
patterns.

Key words:
Lactic acid bacteria, API technique, PCR, FT-IR.

6
 ‫‪ .1‬اﻟﻤﻘﺪﻣﺔ‬
‫ﺗﻤﺘﻠﻚ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ‪ (LAB) Lactic Acid Bacteria‬أهﻤﻴ ًﺔ آﺒﻴ ﺮة ﺑﺎﻋﺘﺒﺎره ﺎ ﻣﻔﻴ ﺪة‬
‫وواﺳ ﻌﺔ اﻻﻧﺘ ﺸﺎر ﻓ ﻲ اﻟﺒﻴﺌ ﺔ‪ .‬وﻗ ﺪ ُﺑ ِﺪئ ﺑﺎﺳ ﺘﺨﺪام ه ﺬﻩ اﻟﺒﻜﺘﻴﺮﻳ ﺎ ﻹﻧﺘ ﺎج اﻟﺤﻤﻮﺿ ﺔ أﺛﻨ ﺎء ﺗ ﺼﻨﻴﻊ‬
‫ﻣﻨﺘﺠﺎت اﻷﻟﺒﺎن اﻟﻤﺘﺨﻤﺮة ﻗﺒﻞ اﻟﺘﻌﺮف ﻋﻠﻴﻬﺎ‪ ،‬ﺣﻴﺚ آﺎن اﻟﺤﻠﻴﺐ ُﻳﺘﺮك ﻓ ﻲ درﺟ ﺔ ﺣ ﺮارة اﻟﻐﺮﻓ ﺔ‬
‫ﻋﺪة ﺳﺎﻋﺎت ﻳﺘﻢ ﺧﻼﻟﻬﺎ ﺗﻜﺎﺛﺮ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠ ﺒﻦ اﻟﻤﻮﺟ ﻮدة ﻓﻴ ﻪ ﻋ ﺎدة‪ ،‬ﺛ ﻢ ﻳ ﺴﺘﺨﺪم ﻓ ﻲ ﺗ ﺼﻨﻴﻊ‬
‫اﻷﻟﺒﺎن واﻷﺟﺒﺎن اﻟﻤﺘﺨﻤﺮة‪ .‬اﺳﺘﻤﺮ اﻟﻌﻤﻞ ﻋﻠﻰ هﺬا اﻟﻨﺤﻮ ﺣﺘﻰ أﺛﺒ ﺖ اﻟﻌﺎﻟﻤ ﺎن ‪ Baily‬و‪Hammer‬‬

‫ﻋﺎم ‪ 1919‬أن اﻟﺒﺎدئ اﻟﻮاﺟﺐ اﺳﺘﺨﺪاﻣﻪ ﻓﻲ إﻧﺘﺎج ﻣﺸﺘﻘﺎت ﻟﺒﻨﻴﺔ ذات ﻃﻌﻢ وﻧﻜﻬﺔ ﻣﺮﻏﻮﺑﻴﻦ ﻳﺠﺐ‬
‫أن ﻳﺤﺘﻮي ﻋﻠﻰ ﻧﻮﻋﻴﻦ ﻣﻦ اﻟﺒﻜﺘﻴﺮﻳﺎ‪ :‬اﻷول ﻳﻨﺘﺞ ﺣﻤﺾ اﻟﻠﺒﻦ‪ ،‬ﻓﻲ ﺣﻴﻦ ﻳﻜﻮن اﻟﺜ ﺎﻧﻲ ﻗ ﺎدرًا ﻋﻠ ﻰ‬
‫إﻧﺘﺎج اﻷﺣﻤﺎض اﻟﻄﻴﺎرة اﻟﺘﻲ ﺗﻤﻨﺢ اﻟﻄﻌﻢ اﻟﻤﻤﻴ ﺰ واﻟﻨﻜﻬ ﺔ )‪ .(Ayad et al., 2003‬ﺑﺎﻹﺿ ﺎﻓﺔ إﻟ ﻰ‬
‫دور اﻟﺒﺎدﺋﺎت ﻓﻲ إﻋﻄ ﺎء اﻟﻘ ﻮام اﻟﻤﻨﺎﺳ ﺐ ﻟﻤﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟﻤﺘﺨﻤ ﺮة ﻓﺈﻧﻬ ﺎ ﺗﺮﻓ ﻊ اﻟﻘﻴﻤ ﺔ اﻟﻐﺬاﺋﻴ ﺔ‬
‫واﻟﺼﺤﻴﺔ ﻟﻬﺬﻩ اﻟﻤﻨﺘﺠﺎت )‪.(Aslim and Beyatli, 2004‬‬
‫وﺣﻴ ﺚ أن اﻟﻐ ﺬاء اﻟﻤﻨ ﺘﺞ ﺑﺈﺿ ﺎﻓﺔ ﻋ ﺼﻴﺎت ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ﻳ ﺸﻜﻞ ﺟ ﺰءًا ﻣ ﻦ اﻟﻐ ﺬاء‬
‫اﻟﻴ ﻮﻣﻲ ﻟﻠﻤ ﺴﺘﻬﻠﻚ )ﻣﺜ ﻞ‪ :‬اﻟﺠﺒﻨ ﺔ‪ ،‬اﻟﺰﺑ ﺪة(‪ ،‬وه ﻮ ﻳﻤﺘﻠ ﻚ ﺻ ﻔﺎت ﻏﺬاﺋﻴ ﺔ إﻳﺠﺎﺑﻴ ﺔ‪ ،‬ﻓ ﺈن اﺳ ﺘﺨﺪام‬
‫ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﻟﻦ ﻳﺜﻴﺮ ﻗﻠﻘًﺎ ﻋﻤﻴﻘًﺎ‪ .‬ورﻏﻢ ذﻟﻚ ﻓﻘﺪ ﻳﺴﺘﻐﺮب اﻟﻤﺴﺘﻬﻠﻚ ﻣﻦ اﻟﺤﻘﻴﻘﺔ اﻟﺘﻲ ﺗﺆآﺪ‬
‫اﺳﺘﺨﺪام اﻟﺒﻜﺘﻴﺮﻳ ﺎ ﻓ ﻲ اﻟﻤﻨﺘﺠ ﺎت اﻟﻐﺬاﺋﻴ ﺔ وﺧﺎﺻ ﺔ ﻋﻨ ﺪ ﻣﻌﺮﻓ ﺔ اﻟﻌ ﺪد اﻟﻬﺎﺋ ﻞ ﻣ ﻦ اﻟﺒﻜﺘﻴﺮﻳ ﺎ اﻟﺤﻴ ﺔ‬
‫اﻟﺬي ﻳﺘﻨﺎوﻟﻪ ﻣﻊ آﻞ ﻟﻘﻤﺔ!‬
‫ﺗﻢ اﻟﺘﻌﺮف ﻋﻠﻰ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﻓﻲ ﺑﺪاﻳﺔ اﻟﻘﺮن اﻟﺘﺎﺳﻊ ﻋﺸﺮ واﻋﺘﺒﺮت ﺑﺄﻧﻬ ﺎ "اﻟﻜﺎﺋﻨ ﺎت‬
‫اﻟﺤﻴ ﺔ اﻟﺘ ﻲ ﺗ ﺆدي إﻟ ﻰ ﺣﻤﻮﺿ ﺔ اﻟﺤﻠﻴ ﺐ"‪ ،‬أﻣ ﺎ اﻟﺒﻜﺘﻴﺮﻳ ﺎ اﻟﻌ ﺼﻮﻳﺔ اﻟﻤ ﺴﺆوﻟﺔ ﻋ ﻦ ﺗﺨﻤ ﺮ اﻟﻠ ﺒﻦ‬
‫اﻟﺒﻠﻐﺎري ﻓﻘﺪ ﺟﺮى اآﺘﺸﺎﻓﻬﺎ ﻓﻲ ﺑﺪاﻳﺔ اﻟﻘﺮن اﻟﻌ ﺸﺮﻳﻦ‪ .‬وﻗ ﺪ وﺟ ﺪ اﻟﻌ ﺎﻟﻢ ‪ Henneberg‬ﻋ ﺎم ‪1904‬‬

‫أن اﻟﺒﻜﺘﻴﺮﻳ ﺎ اﻟﺘ ﻲ ﺗﺘ ﺴﺒﺐ ﻓ ﻲ ﺣﻤﻮﺿ ﺔ اﻟﻠ ﺒﻦ ﺗ ﺸﺒﻪ أﻧ ﻮاع ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ اﻟﻤﻮﺟ ﻮدة ﻓ ﻲ‬
‫اﻟﺘﺮﺑ ﺔ‪ .‬أﻣ ﺎ ‪ Grigoroff‬ﻓﻘ ﺪ ﻋ ﺰل ﻣ ﻦ اﻟﺤﻠﻴ ﺐ‪ ،‬ﻋ ﺎم ‪ ،1905‬آﺎﺋﻨ ﺎت دﻗﻴﻘ ﺔ ﺗﺨﺘﻠ ﻒ ﺑﺄﺷ ﻜﺎﻟﻬﺎ‬
‫اﻟﻤﺠﻬﺮﻳﺔ ﻓﻤﻨﻬﺎ اﻟﻌﺼﻮي ‪ rods‬وﻣﻨﻬﺎ اﻟﻜﺮوي ‪.(Gobbetti, 2001) cocci‬‬

‫‪ 1.1‬ﺗﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪:‬‬

‫ﻗﺎم ﺟﻮزﻳﻒ ﻟﻴﺴﺘﺮ ﻓﻲ ﻋ ﺎم ‪ 1873‬ﺑ ﺄول اﻟﺪراﺳ ﺎت ﻋ ﻦ اﻟ ـ ‪ Lactococci‬ﺣﻴ ﺚ ﺣ ﺎول إﺛﺒ ﺎت‬
‫ﻧﻈﺮﻳ ﺔ ﺑﺎﺳ ﺘﻮر ذات اﻟ ﺼﻠﺔ ﺑ ﺎﻟﺘﻐﻴﺮات اﻟﺘﺨﻤﺮﻳ ﺔ اﻟﺘ ﻲ ﺗ ﺴﺒﺒﻬﺎ اﻷﺣﻴ ﺎء اﻟﺪﻗﻴﻘ ﺔ‪ .‬ﺗﻤﻜ ﻦ ﻟﻴ ﺴﺘﺮ ﻓ ﻲ‬

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‫ﺗﺠﺎرﺑﻪ اﻟﺘﻲ اﺳﺘﺨﺪﻣﺖ اﻟﺤﻠﻴﺐ اﻟﻤﻐﻠﻲ آﻮﺳﻂ ﻣﻐ ٍﺬ‪ ،‬ﻣﻦ اﻟﺤﺼﻮل ﻋﻠﻰ أول زراﻋﺔ ﺑﻜﺘﻴﺮﻳﺔ ﻧﻘﻴﺔ؛‬
‫ﻼ‪:‬‬
‫ووﺻﻔﻬﺎ ﻗﺎﺋ ً‬
‫"ﻋﻠﻴﻨ ﺎ اﻻﻋﺘ ﺮاف ﺑﺄﻧﻨ ﺎ ﻧﺘﻌﺎﻣ ﻞ هﻨ ﺎ ﻣ ﻊ ﺟ ﻨﺲ ﺑﻜﺘﻴ ﺮي واﺣ ﺪ ﻓﻘ ﻂ ُﻳﻈْ ِﻬ ﺮ ﺧﻮاﺻ ﻪ اﻟ ﺸﻜﻠﻴﺔ‬
‫واﻟﻔﻴﺰﻳﻮﻟﻮﺟﻴ ﺔ‪ ،‬وأﻗﺘ ﺮح ﺗ ﺴﻤﻴﺘﻪ ‪ .Bacterium lactis‬أﻗ ﻮم ﺑﻬ ﺬا ﺑ ﺘﺤﻔﻆ ﻣﻌﺘﻘ ﺪًا أﻧ ﻪ ﺣﺘ ﻰ ه ﺬا‬
‫اﻟﺘﺎرﻳﺦ‪ ،‬ﻟﻢ ﻳ ﺘﻢ اآﺘ ﺸﺎف ﺑﻜﺘﻴﺮﻳ ﺎ ﺗﺘﻤﺘ ﻊ ﺑ ﻨﻔﺲ ه ﺬﻩ اﻟﻤﻮاﺻ ﻔﺎت‪ .‬آﻤ ﺎ أﻋﺘﻘ ﺪ أﻧﻬ ﺎ ﻟﻴ ﺴﺖ اﻟﺒﻜﺘﻴﺮﻳ ﺎ‬
‫ﺳ ﱢﻤﻴَﺖ ﻻﺣﻘ ًﺎ‬
‫اﻟﻮﺣﻴﺪة اﻟﺘ ﻲ ﺗﻘ ﻮم ﺑﻌﻤﻠﻴ ﺔ اﻟﺘﺨﻤ ﺮ اﻟﻠﺒﻨ ﻲ"‪ .‬وﺗﺠ ﺪر اﻹﺷ ﺎرة إﻟ ﻰ أن ه ﺬﻩ اﻟﺒﻜﺘﻴﺮﻳ ﺎ ُ‬
‫"‪."Streptococcus lactis‬‬

‫اﻟﻄﺮاﺋﻖ اﻟﺘﻘﻠﻴﺪﻳﺔ ﻟﺘﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪:‬‬

‫ﻗﺎم ‪ (1919) Orla-Jensen‬ﺑﻮﺿﻊ أﺳﺲ ﺗ ﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ )اﻟﺘ ﻲ ُﺗ ﺴْ َﺘﺨْﺪم ﺣﺘ ﻰ‬
‫وﻗﺘﻨﺎ اﻟﺤﺎﻟﻲ‪ ،‬رﻏﻢ إدﺧﺎل ﺑﻌﺾ اﻟﺘﻌﺪﻳﻼت(‪ ،‬ﻣﻌﺘﻤ ﺪًا ﻋﻠ ﻰ اﻷﺳ ﺎس اﻟ ﺸﻜﻠﻲ )آ ﺮوي‪ ،‬ﻋ ﺼﻮي(‬
‫وﻧﻤﻂ اﻟﺘﺠﻤﻊ )ﻣﻔﺮد‪ ،‬ﺛﻨﺎﺋﻲ‪ ،‬ﺳﻠﺴﻠﻲ(‪ ،‬وﻧﻤﻂ ﺗﺨﻤﻴﺮ ﺳﻜﺮ اﻟﻐﻠﻮآ ﻮز )ﻣﺘﺠ ﺎﻧﺲ‪ ،‬ﻏﻴ ﺮ ﻣﺘﺠ ﺎﻧﺲ(‪،‬‬
‫إﻟﻰ ﺟﺎﻧﺐ دراﺳﺔ ﻧﻤﻮ هﺬﻩ اﻟﺒﻜﺘﻴﺮﻳﺎ ﻓﻲ درﺟﺎت ﺣ ﺮارة ﻣﺨﺘﻠﻔ ﺔ ﻻﺳ ﻴﻤﺎ ‪˚10‬م و‪˚45‬م )أﺑ ﻮ ﻳ ﻮﻧﺲ‬
‫وزﻣﻼؤهﺎ‪ .(2007 ،‬وﺑﻨﺎ ًء ﻋﻠﻰ ذﻟﻚ‪َ ،‬ﻗﺴﱠﻢ ‪ Orla-Jensen‬ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ إﻟ ﻰ أرﺑﻌ ﺔ أﺟﻨ ﺎس‬
‫هﻲ‪:‬‬
‫‪.Streptococcus ،Pediococcus ،Leuconostoc ،Lactobacillus‬‬
‫اﻟ ـ‬ ‫وﻋﻨﺪ إﺟﺮاء إﻋﺎدة ﺗﺤﻘﻴ ﻖ ﺷ ﺎﻣﻠﺔ‪ ،‬اﻗﺘ ﺮح ‪ Schleifer‬وزﻣ ﻼؤﻩ )‪َ (1985‬ﻓ ﺼْﻞ‬
‫‪ Enterococci‬ﻋ ﻦ اﻟ ـ ‪ Streptococci‬اﻟﺤﺎﻟ ﺔ ﻟﻠ ﺪم‪ ،‬وﻣ ﻦ ﺛ ﻢ ﺗﻤ ﺖ ﺗ ﺴﻤﻴﺘﻬﺎ ﺑﺎﺳ ﻢ ﺟ ﻨﺲ ﺟﺪﻳ ﺪ ه ﻮ‬
‫"‪ .(Naser et al., 2005) "Lactococcus‬وﻳﻮﺿ ﺢ اﻟ ﺸﻜﻞ ‪ 1‬ﺷ ﺠﺮة اﻟﺘﻄ ﻮر ‪phylogenetic‬‬

‫ﻟﺒﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪ .‬ﻣﻦ ﻧﺎﺣﻴﺔ أﺧﺮى‪ ،‬ﻳﻨﺒﻐﻲ ﻣﺮاﻋﺎة وﺟﻮد اﺧﺘﻼﻓﺎت ﻓﻲ اﻟﺤ ﺪود اﻟﻔﺎﺻ ﻠﺔ ﺑ ﻴﻦ‬
‫‪ .(Williams‬وﻗ ﺪ ﺣ ﺪد ‪ Orla-Jensen‬اﻷﻧ ﻮاع‬ ‫‪and‬‬ ‫‪Coliins,‬‬ ‫اﻷﺟﻨ ﺎس )‪1990‬‬

‫‪ Thermobacterium bulgaricum‬اﻟﻤﺴﻤﺎة ﺣﺎﻟﻴًﺎ ‪Tharmaraj and ) Lactobacillus bulgaricus‬‬

‫‪.(Shah, 2003‬‬

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‫اﻟﺸﻜﻞ ‪ .1‬ﻣﺨﻄﻂ ﺗﻮﺿﻴﺤﻲ ﻟﺘﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‬

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‫اﻋﺘﻤ ﺎدًا ﻋﻠ ﻰ اﻟﺪراﺳ ﺔ اﻟﻬﺎﻣ ﺔ اﻟﺘ ﻲ ﻗ ﺎم ﺑﻬ ﺎ ‪ Patrick Tailliez‬ﻋ ﻦ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ‬
‫اﻟﻤﺘﻀﻤﻨﺔ ﻟﻠـ ‪ ،Lactococci‬ﻓﻘ ﺪ اﻗﺘ ﺮح أن ﺗﻜ ﻮن ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ﻗ ﺪ ﻇﻬ ﺮت ﻗﺒ ﻞ اﻟﺒﻜﺘﻴﺮﻳ ﺎ‬
‫ﺟ ﺪت ﻓ ﻲ رﺳ ﻮﺑﻴﺎت‬
‫اﻟﺰرﻗﺎء ‪ Cyanobacteria‬اﻟﺘﻲ ﺗﻘﻮم ﺑﻌﻤﻠﻴﺔ اﻟﺘﺮآﻴ ﺐ اﻟ ﻀﻮﺋﻲ‪ .‬وﺑﻤ ﺎ أﻧﻬ ﺎ ُو ِ‬
‫ﻋﻤﺮهﺎ ‪ 2.75‬ﺑﻠﻴﻮن ﺳﻨﺔ‪ ،‬ﻓ ُﻴﺤْ َﺘ َﻤﻞ أن ﺗﻜﻮن ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﻗﺪ ﻇﻬﺮت ﻗﺒﻞ ‪ 3‬ﺑﻠﻴﻮن ﺳﻨﺔ ﻣ ﻦ‬
‫ﺴﺮ ﺗﻜﻴﻔﻬ ﺎ اﻟ ﻀﻌﻴﻒ‬
‫اﺣﺘﻮاء اﻟﻐﻼف اﻟﺠﻮي ﻋﻠﻰ اﻟﻜﻤﻴﺔ اﻟﻜﺎﻓﻴﺔ ﻣﻦ اﻷآﺴﺠﻴﻦ‪ ،‬وهﺬا ﻳﻤﻜﻦ أن ﻳﻔ ﱢ‬
‫ﻣ ﻊ اﻟﺒﻴﺌ ﺔ اﻟﻬﻮاﺋﻴ ﺔ‪ .‬ﺑﺎﻟﻤﻘﺎﺑ ﻞ‪ ،‬أﺷ ﺎر ‪ Tailliez‬إﻟ ﻰ أن اﻟ ـ ‪ Lactococci‬ﻓ ﻲ ﻃﺮﻳﻘﻬ ﺎ ﻟﻠﺘﺤ ﻮل إﻟ ﻰ‬
‫اﻟﺘﻨﻔﺲ ﺑﺸﻜﻞ واﺿﺢ )‪.(2001‬‬
‫اﻟﻄﺮاﺋﻖ اﻟﺤﺪﻳﺜﺔ ﻟﺘﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪:‬‬
‫ﺗﻌﺘﻤﺪ اﻟﻄﺮاﺋﻖ اﻟﺤﺪﻳﺜﺔ ﻟﺘﺼﻨﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﻋﻠ ﻰ ﻣﺤﺘ ﻮى ه ﺬﻩ اﻷﺧﻴ ﺮة ﻣ ﻦ اﻟﻤ ﺎدة‬
‫اﻟﻮراﺛﻴ ﺔ واﻷﺣﻤ ﺎض اﻟﻨﻮوﻳ ﺔ ‪.(Ehrmann and Vogel, 2005; Settanni et al., 2005) DNA‬‬
‫وﺗﺘﻤﺘﻊ هﺬﻩ اﻟﻄﺮاﺋ ﻖ ﺑﺪﻗ ﺔ أآﺒ ﺮ ﻓ ﻲ ﺗﺤﺪﻳ ﺪ اﻷﻧ ﻮاع وﺗﺤ ﺖ اﻷﻧ ﻮاع اﻟﺘ ﻲ ﺗﻤﺘﻠ ﻚ ﺗﻨﻮﻋ ًﺎ واﺧﺘﻼﻓ ًﺎ‬
‫آﺒﻴ ﺮﻳْﻦ ﻓ ﻲ اﻟﻤ ﺎدة اﻟﻮراﺛﻴ ﺔ ﻳ ﺴﺎﻋﺪان ﻓ ﻲ ﺗﻤﻴﻴﺰه ﺎ ﻋ ﻦ ﺑﻌ ﻀﻬﺎ )‪ .(Bae et al., 2005‬وﺗﻘ ﻮم‬
‫اﻟﻄﺮﻳﻘ ﺔ اﻟﺤﺪﻳﺜ ﺔ ﻋﻠ ﻰ ﺗﺮﺣﻴ ﻞ اﻟﻤ ﺎدة اﻟﻮراﺛﻴ ﺔ ﻟﻠﺒﻜﺘﻴﺮﻳ ﺎ ﻋﻠ ﻰ هﻼﻣ ﺔ ﻣﻮﺿ ﻮﻋﺔ ﺿ ﻤﻦ ﺣﻘ ﻞ‬
‫آﻬﺮﺑ ﺎﺋﻲ )‪ (gel-electrophoresis‬ﺑﻌ ﺪ اﻟﻘﻴ ﺎم ﺑﻌﻤﻠﻴ ﺔ ﺗ ﻀﺨﻴﻢ ﻣ ﻮرﺛﻲ ﻟﻤﻮرﺛ ﺔ ﻣﺤﺎﻓﻈ ﺔ ﺿ ﻤﻦ‬
‫اﻟﺠﻨﺲ‪ .‬وﻳﻤﻜﻦ ﺗﻄﺒﻴﻖ هﺬﻩ اﻟﻄﺮﻳﻘﺔ ﻋﻠﻰ أﻧﻮاع ﻣﺘﻌﺪدة ﻣﻦ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﺷﺮﻳﻄﺔ اﺳ ﺘﺨﺪام‬
‫ُﻣ َﺮﺋﱢﺴﺎت ﻣﺨﺘﻠﻔﺔ وﻣﻨﺎﺳﺒﺔ )‪.(Picozzi et al., 2006‬‬
‫اﺳْ ُﺘﺨْ ِﺪﻣَﺖ ه ﺬﻩ اﻟﻄﺮﻳﻘ ﺔ ﻣ ﻦ أﺟ ﻞ ﺗ ﺼﻨﻴﻒ ‪ Lb. casei subsp. Casei‬اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ أﻧ ﻮاع‬
‫ﻣﺨﺘﻠﻔ ﺔ ﻣ ﻦ اﻷﺟﺒ ﺎن اﻷوروﺑﻴ ﺔ‪ ،‬وآ ﺬﻟﻚ ﻟﺘ ﺼﻨﻴﻒ ‪ Lactococcus lactis‬و ‪Lactobacillus‬‬

‫‪ plantarum‬اﻟﻤﻌﺰوﻟﺘﻴْﻦ ﻣﻦ اﻟﺤﻠﻴﺐ اﻟﺨﺎم اﻟ ُﻤﺴْ َﺘﺨْﺪَم ﻹﻧﺘ ﺎج ﺟ ﺒﻦ اﻟﻜ ﺎﻣﻤﺒﻴﺮ اﻟﻔﺮﻧ ﺴﻲ ) ‪Mangin‬‬

‫‪ .(et al., 1999‬ﺗﻤﻜ ﻦ اﻟﺒ ﺎﺣﺜﻮن اﻋﺘﻤ ﺎدًا ﻋﻠ ﻰ ﻃﺮﻳﻘ ﺔ اﻟ ـ ‪ PCR‬ﻣ ﻦ اﻟﺘﻤﻴﻴ ﺰ ﺑ ﻴﻦ اﻷﻧ ﻮاع ‪Lb.‬‬

‫‪ acidophilus‬و‪ L. gasseri‬و‪ L. johndonii‬وهﻲ ﺟﻤﻴﻌﻬﺎ أﻧﻮاع ﻋﺼﻮﻳﺔ اﻟﺸﻜﻞ وﻣﻔﻴﺪة‪ ،‬آﺎﻧﺖ ﻗﺪ‬
‫ﺻﻨﱢﻔَﺖ ﻗﺪﻳﻤًﺎ ﻋﻠﻰ أﻧﻬﺎ ﻣﻨﺘﻤﻴﺔ إﻟﻰ ﻣﺠﻤﻮﻋﺔ ‪ acidophilus‬وذﻟﻚ اﻋﺘﻤﺎدًا ﻋﻠ ﻰ اﻟﺘﺤﻠﻴ ﻞ اﻟﺒﺮوﺗﻴﻨ ﻲ‬
‫ُ‬
‫)‪ .(Randazzo et al., 2005‬وﺑﺎﺳﺘﺨﺪام اﻟـ ‪ ،PCR‬ﺗﻢ ﺟﻤﻊ اﻟﻨﻮﻋﻴﻦ ‪ Lb. casei‬و ‪Lb. paracasei‬‬

‫ﺳ ﻤِﻲ ‪ Lb. casei‬ﻧﻈ ﺮًا ﻟﺘﻄ ﺎﺑﻖ ﻣﺤﺘﻮﻳﻬﻤ ﺎ ﻣ ﻦ اﻟ ـ ‪DNA‬‬

‫‪ subsp. paracasei‬ﻓ ﻲ ﻧ ﻮع واﺣ ﺪ ُ‬
‫)ﻣﺎدﺗﻬﻤﺎ اﻟﻮراﺛﻴ ﺔ(‪ ،‬ﻋﻠﻤ ًﺎ أن اﻟﺘ ﺼﻨﻴﻒ اﻟ ﺴﺎﺑﻖ اﻟ ﺬي آ ﺎن ﻳﻌﺘﻤ ﺪ ﻋﻠ ﻰ ﺗﺤﻠﻴ ﻞ اﻟﺠ ﺪار اﻟﺒﺮوﺗﻴﻨ ﻲ‬
‫ﻟﻠﺒﻜﺘﻴﺮﻳﺎ هﻮ اﻟﺬي ﻓﺼﻠﻬﻤﺎ إﻟﻰ ﻧﻮﻋﻴْﻦ )‪.(Corsetti et al., 2003‬‬

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‫ﺳﺎهﻢ اﻟﺘﺼﻨﻴﻒ اﻟﺠﺰﻳﺌﻲ ﻟﻠـ ‪ Lactococci‬ﻓﻲ اآﺘﺸﺎف وﺗﻤﻴﻴﺰ أﻧﻮاع ﺟﺪﻳﺪة ﺑﺎﻹﺿﺎﻓﺔ إﻟﻰ ‪L.‬‬

‫ﺨ ﺼَﺖ‬
‫ﺷِ‬
‫‪ lactis‬وه ﻲ‪ L. graviae :‬و‪ L. raffinolactis‬و‪ L. plantarum‬و‪ .L. piscium‬ﺣﻴ ﺚ ُ‬
‫‪ Lactococcus graviae‬ﻋﻠﻰ أﻧﻬﺎ اﻟﻌﺎﻣﻞ اﻟﻤﺴﺒﺐ ﻻﻟﺘﻬﺎب اﻟﻀﺮع ﻋﻨﺪ اﻷﺑﻘﺎر‪.‬‬
‫ل اﻟﻨ ﻮع ‪ L. lactis‬ﻣ ﻦ اﻟﺒ ﺸﺮ ﻋﻨ ﺪ إﺻ ﺎﺑﺔ اﻟﺠﻬ ﺎز اﻟﺒ ﻮﻟﻲ واﻟﺘﻬ ﺎب‬
‫ﻋ ِﺰ َ‬
‫وﻓ ﻲ ﺣ ﺎﻻت ﻧ ﺎدرة‪ُ ،‬‬
‫اﻟﺠﺮوح وﻣﻦ ﻣﺮﺿﻰ اﻻﻟﺘﻬﺎب اﻟﺸﻐﺎﻓﻲ‪.‬‬
‫أﺟﻨﺎس وأﻧﻮاع ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪:‬‬
‫ﺗﺪل ﻋﺒﺎرة ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﺗ ﺼﻨﻴﻔﻴﺎً‪ ،‬ﻋﻠ ﻰ ﻣﺠﻤﻮﻋ ﺔ ﻣ ﻦ اﻟﺒﻜﺘﻴﺮﻳ ﺎ اﻟﺘ ﻲ ﺗﺘﻤﺘ ﻊ ﺑﺎﻟﺨ ﺼﺎﺋﺺ‬
‫اﻟﻤﺸﺘﺮآﺔ اﻟﺘﺎﻟﻴﺔ )‪:(Davidson et al., 1996‬‬
‫ﻣﻮﺟﺒ ﺔ اﻟﻐ ﺮام‪ ،‬ﻏﻴ ﺮ ﻣﺘﺒﻮﻏ ﺔ‪ ،‬ﺳ ﺎﻟﺒﺔ اﻟﻜﺎﺗ ﺎﻻز‪ ،‬ﺧﺎﻟﻴ ﺔ ﻣ ﻦ اﻟ ﺴﻴﺘﻮآﺮوﻣﺎت‪ ،‬إﻻ أﻧﻬ ﺎ ذات ﺗﺤﻤ ﻞ‬
‫ﺞ اﻟﺮﺋﻴ ﺴﻲ اﻟﻨﻬ ﺎﺋﻲ ﻟﺘﺨﻤ ﺮ اﻟ ﺴﻜﺮ اﻟ ﺬي‬
‫هﻮاﺋﻲ‪ ،‬وذات ﺗﺤﻤﻞ ﻟﻠﺤﻤﻮﺿﺔ‪ ،‬و ُﻳﻌَﺪ ﺣﻤﺾ اﻟﻠﺒﻦ اﻟﻨ ﺎﺗ َ‬
‫ﺗﻘﻮم ﺑﻪ هﺬﻩ اﻟﺒﻜﺘﻴﺮﻳﺎ‪.‬‬

‫ﺗﺘﻀﻤﻦ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ اﻟﻨﺎﺗﺠﺔ ﻋﻦ ﺗﺨﻤﺮات اﻷﻏﺬﻳﺔ اﻷﺟﻨﺎس اﻟﺘﺎﻟﻴﺔ‪:‬‬

‫‪Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc,‬‬
‫‪Oenococcus, Pediococcus, Streptococcus, Tetragenococcus, Weissella, Vogococcus‬‬
‫)‪ .(Konstantinidis and Tiedje, 2005‬وﻗ ﺪ ﺟﻤ ﻊ دﻟﻴ ﻞ ﺑﻴﺮﺟ ﻲ ‪ ،Bergey's Manual‬ﻓ ﻲ‬
‫ﻃﺒﻌﺘ ﻪ اﻟ ﺴﺎﺑﻌﺔ ﻟﻌ ﺎم ‪ ،1957‬ﻋ ﺎﺋﻠﺘﻲْ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ‪) Streptococceae‬ذات اﻟ ﺸﻜﻞ‬
‫اﻟﺒﻴ ﻀﻮي أو اﻟﻜ ﺮوي( و‪) Lactobacilleae‬ذات اﻟ ﺸﻜﻞ اﻟﻌ ﺼﻮي( ﻓ ﻲ ﻋﺎﺋﻠ ﺔ واﺣ ﺪة ﺳ ﻤﺎهﺎ‬
‫‪ .Lactobacillaceae‬إﻻ أﻧ ﻪ ﻣ ﺎ ﻟﺒ ﺚ أن ﻋ ﺎد وﻓ ﺼﻠﻬﻤﺎ إﻟ ﻰ ﻋ ﺎﺋﻠﺘﻴْﻦ هﻤ ﺎ ‪Streptococcaceae‬‬

‫و‪ Lactobacillaceae‬وذﻟ ﻚ ﻓ ﻲ ﻃﺒﻌﺘ ﻪ اﻟﺜﺎﻣﻨ ﺔ ﻋ ﺎم ‪) 1974‬أﺑ ﻮ ﻳ ﻮﻧﺲ وزﻣﻼؤه ﺎ‪ .(2007 ،‬وﻗ ﺪ‬

‫ﺗﺒ ﻴﻦ ﻓ ﻲ ﻣﻨﺘ ﺼﻒ اﻟﺜﻤﺎﻧﻴﻨ ﺎت أن اﻟ ـ ‪ S. thermophilus‬ه ﻮ اﻟﻨ ﻮع اﻟﻮﺣﻴ ﺪ ﻣ ﻦ اﻟﺠ ﻨﺲ‬
‫‪ Streptococcus‬اﻟﺬي ﻟﻪ ﻋﻼﻗﺔ ﺑﺘﺨﻤﺮ اﻟﻐﺬاء‪ ،‬ﺣﻴﺚ ﻳﻨﺘﻤﻲ هﺬا اﻟﺠ ﻨﺲ إﻟ ﻰ اﻟ ـ ‪ LAB‬ﻻﺷ ﺘﺮاآﻪ‬
‫ﻣﻌﻬ ﺎ ﺑ ﺒﻌﺾ اﻟﻤﻼﻣ ﺢ اﻟﻨﻤﻄﻴ ﺔ؛ ﻣﺜ ﻞ اﻻﺳ ﺘﻘﻼب اﻟﺘﺨﻤ ﺮي وإﻧﺘ ﺎج ﺣﻤ ﺾ اﻟﻠ ﺒﻦ‪ ،‬رﻏ ﻢ أن ه ﺬﻩ‬
‫اﻟﺒﻜﺘﻴﺮﻳ ﺎ ﻻ ﺗ ﺮﺗﺒﻂ ﻣ ﻊ اﻟ ـ ‪ LAB‬ﻣ ﻦ اﻟﻨﺎﺣﻴ ﺔ اﻟﺘﻄﻮرﻳ ﺔ )اﻟﺘ ﺄرﻳﺦ اﻟﻌﺮﻗ ﻲ( ‪phylogenetically‬‬

‫وﺗﺴﺘﺨﺪم ﻃﺮﻳﻘﺔ وﺣﻴﺪة ﻻﺳﺘﻘﻼب اﻟﺴﻜﺮ‪ .‬آﻤﺎ ﺗﻢ ﺿﻢ اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﻤﺴﺒﺒﺔ ﻟﻠﺘﻘﻴﺤﺎت إﻟﻰ هﺬا اﻟﺠ ﻨﺲ‬
‫ذاﺗ ﻪ‪ .‬وآ ﺬﻟﻚ أﻇﻬ ﺮ دﻟﻴ ﻞ ﺑﻴﺮﺟ ﻲ ﻓ ﻲ ﻃﺒﻌﺘ ﻪ اﻟﺘﺎﺳ ﻌﺔ ﻋ ﺎم ‪ 1986‬أﺟﻨﺎﺳ ًﺎ أﺧ ﺮى ﻏﻴ ﺮ‬
‫اﻟ ـ ‪ Streptococcus‬ه ﻲ‪Aerococcus, Lactobacillus, Leuconostoc, Pediococcus:‬؛ وﻳ ﻀﻢ‬

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‫آﻞ ﺟﻨﺲ ﺗﺤﺖ أﺟﻨﺎس ﻋﺪﻳ ﺪة‪ .‬إﻻ أن اﻟﻄﺒﻌ ﺎت اﻷﺣ ﺪث ﻟ ﺪﻟﻴﻞ ﺑﻴﺮﺟ ﻲ ﻗ ﺴﻤﺖ ه ﺬﻩ اﻟﻤﺠﻤﻮﻋ ﺎت‬
‫إﻟ ﻰ أﺟﻨ ﺎس ﻣ ﺴﺘﻘﻠﺔ ﻓﻈﻬ ﺮ ﺟ ﻨﺲ اﻟ ـ ‪ Enterococcus‬وﺟ ﻨﺲ اﻟ ـ ‪ Lactococcus‬وﺟ ﻨﺲ اﻟ ـ‬
‫‪) Vagococcus‬أﺑﻮ ﻳﻮﻧﺲ وزﻣﻼؤهﺎ‪ .(2007 ،‬وﺑﺴﺒﺐ ﺑﻌﺾ اﻷﺧﻄﺎء واﻟﻤ ﺸﺎآﻞ ﻓ ﻲ اﻟﺘ ﺼﻨﻴﻒ‪،‬‬
‫ﻼ أن اﻟﻨ ﻮع ‪ E. solitarius‬آ ﺎن أﺷ ﺪ‬
‫ﺟﺪ ﻣ ﺜ ً‬
‫أﻋﻴﺪ ﺗﺼﻨﻴﻒ ﺑﻌﺾ أﻧﻮاع اﻟﺠﻨﺲ ‪ Enterococcus‬ﻓ ُﻮ ِ‬
‫ﻗﺮاﺑ ًﺔ إﻟﻰ اﻟﺠﻨﺲ ‪.(Eaton and Gasson, 2001; Giraffa, 2002) Tetragenococcus‬‬
‫‪ 4.1‬ﺑﻴﺌﺔ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‪:‬‬
‫ﺗﺨﺘﻠﻒ ﻣﻨﺘﺠﺎت اﻷﻟﺒﺎن اﻟﻤﺼﻨﻌﺔ ﺑ ﺸﻜﻞ ﺗﻘﻠﻴ ﺪي ﻋ ﻦ ﺗﻠ ﻚ اﻟﻤ ﺼﻨﻌﺔ ﺗﺠﺎرﻳ ًﺎ ﺑ ﺎﻟﻄﻌﻢ واﻟﻨﻜﻬ ﺔ‬
‫واﻟﻘ ﻮام ﺑ ﺴﺒﺐ اﻟﻔﻠ ﻮرا اﻟﻄﺒﻴﻌﻴ ﺔ ﻟﺒﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ اﻟﻤﺘﻮاﺟ ﺪة ﻓﻴﻬ ﺎ‪ ،‬وه ﻮ ﻣ ﺎ ﺗﻄﻠ ﻖ ﻋﻠﻴ ﻪ‬
‫اﻟﺪراﺳ ﺎت اﻟﺤﺪﻳﺜ ﺔ اﺳ ﻢ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ﺑ ﺪون ﺑ ﺎدئ )‪ .(Kieronczyk et al., 2003‬وه ﻲ‬
‫ﺼ ﱠﻨﻌﺔ ﻏﺎﻟﺒ ًﺎ ﻣ ﻦ ﺣﻠﻴ ﺐ ﻏﻴ ﺮ ﻣﻌﺎﻣ ﻞ‬
‫ﺗﺘﻤﺘﻊ ﺑﺄهﻤﻴﺔ آﺒﻴﺮة ﻧﻈﺮًا ﻟﺘﻮاﺟ ﺪهﺎ ﻓ ﻲ ﻣﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟ ُﻤ َ‬
‫ف )‪.(Beresford et al., 2001‬‬
‫ﺣﺮارﻳًﺎ أو أﻧﻪ ﻣﻌﺎﻣﻞ ﺑﺸﻜﻞ ﻏﻴﺮ آﺎ ٍ‬
‫وﻓﻲ ﺟﺒﻦ اﻟﺸﻴﺪر‪ ،‬ﺗﻌﺪ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ هﻲ اﻟﻤﺴﻴﻄﺮة ﻋﻠﻰ اﻟﻔﻠ ﻮرا اﻟﺒﻜﺘﻴﺮﻳ ﺔ اﻟﻤﺘﻮاﺟ ﺪة‬
‫ﻓﻴ ﻪ ﺑﻌ ﺪ ﺛﻼﺛ ﺔ أﺷ ﻬﺮ ﻣ ﻦ ﺗ ﺼﻨﻴﻌﻪ‪ .‬وﺗ ﺸﻤﻞ ه ﺬﻩ اﻟﺒﻜﺘﻴﺮﻳ ﺎ أﻧﻮاﻋ ًﺎ ﺗﻌ ﻮد إﻟ ﻰ اﻷﺟﻨ ﺎس اﻟﺘﺎﻟﻴ ﺔ‪:‬‬
‫‪ .(Ayad et al., 2001) Lactobacillus, Lactococcus, Leuconostoc, Pediococcus‬ﺗﻨﺘﺞ هﺬﻩ‬
‫اﻟﺒﻜﺘﻴﺮﻳ ﺎ أﺣﻤﺎﺿ ًﺎ ﻣﺨﺘﻠﻔ ﺔ ﻣﺜ ﻞ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ وﺣﻤ ﺾ اﻟﺨ ﻞ‪ ،‬آﻤ ﺎ ﺗﻨ ﺘﺞ ﻣ ﻮاد اﻟﻨﻜﻬ ﺔ آﺎﻟ ﺪي‬
‫أﺳ ﻴﺘﻴﻞ ‪ diacetyl‬واﻷﺳ ﻴﺖ أﻟﺪهﻴ ﺪ ﺑﺤﻴ ﺚ ﺗﻤ ﻨﺢ ه ﺬﻩ اﻷﺧﻴ ﺮة اﻟﻨﻜﻬ ﺔ اﻟﻤﻤﻴ ﺰة ﻟﻤﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن‬
‫اﻟﺘﻘﻠﻴﺪﻳﺔ واﻟﺘﻲ ﺗﻮاﻓﻖ ذوق اﻟﻤﺴﺘﻬﻠﻚ‪ .‬ﺗﻘﻮم هﺬﻩ اﻟﺒﻜﺘﻴﺮﻳ ﺎ أﻳ ﻀًﺎ ﺑﺈﻧﺘ ﺎج اﻷﺳ ﻴﺘﻮﻳﻦ )‪ (acetoin‬ﻣ ﻦ‬
‫ﺗﺨﻤﺮ اﻟﻐﻠﻮآﻮز ﻣﻌﻄﻴ ًﺔ ﺑ ﺬﻟﻚ ﻧﻜﻬ ًﺔ ﻣﻤﻴ ﺰة ﻟ ﺒﻌﺾ اﻟﻤﻨﺘﺠ ﺎت اﻟﻤﺘﺨﻤ ﺮة )ﻋﻠﻤ ًﺎ أن اﻷﺳ ﻴﺘﻮﻳﻦ ه ﻮ‬
‫ﻣﺮآﺐ ﻣﻦ ﻣﺮآﺒﺎت اﻟﻨﻜﻬﺔ وهﻮ اﻟﺬي ﻳﻮﻟﺪ اﻟﺪي أﺳﻴﺘﻴﻞ(‪.‬‬
‫إﻟﻰ ﺟﺎﻧﺐ اﻟﺨﺼﺎﺋﺺ اﻟﺘﻜﻨﻮﻟﻮﺟﻴ ﺔ اﻟﺘ ﻲ ﺗﺘﻤﻴ ﺰ ﺑﻬ ﺎ اﻟﻔﻠ ﻮرا اﻟﻄﺒﻴﻌﻴ ﺔ ﺑﺎﻟﻤﻘﺎرﻧ ﺔ ﻣ ﻊ اﻟﺒﺎدﺋ ﺎت‬
‫اﻟﺘﺠﺎرﻳﺔ‪ ،‬ﻓﺈﻧﻬﺎ ﺗﺘﻤﺘﻊ ﺑﻤﻘﺎوﻣ ﺔ أآﺒ ﺮ ﻟﻠﻤ ﻀﺎدات اﻟﺒﻜﺘﻴﺮﻳ ﺔ )‪ .(Citak et al., 2004‬ه ﺬا ﺑﺎﻹﺿ ﺎﻓﺔ‬
‫ﻀﺠﺔ(‪ ،‬ﻓﻬ ﻲ ﻗ ﺎدرة‬
‫إﻟﻰ إﻣﻜﺎﻧﻴﺔ ﺗﺤﻤﻠﻬ ﺎ ﻟﻠﻤﻌ ﺎﻣﻼت اﻟﺘ ﺼﻨﻴﻌﻴﺔ ﻟﻸﺟﺒ ﺎن )ﺳ ﻮاء اﻟﻄﺎزﺟ ﺔ أو اﻟ ُﻤ َﻨ ﱠ‬
‫ﻋﻠ ﻰ اﻟﻨﻤ ﻮ ﻓ ﻲ ﻧ ﺴﺒﺔ رﻃﻮﺑ ﺔ ‪ %39‬وﻓ ﻲ ﻧ ﺴﺒﺔ ﻣﻠﻮﺣ ﺔ ﻣﺘﺮاوﺣ ﺔ ﻣ ﺎﺑﻴﻦ ‪ 4‬و ‪ %6‬ﻣ ﻦ آﻠﻮرﻳ ﺪ‬
‫اﻟﺼﻮدﻳﻮم ‪ ،NaCl‬ودرﺟﺔ اﻟـ ‪ pH‬ﺗﺘﺮاوح ﻣﺎ ﺑﻴﻦ ‪ 4.9‬و‪) 5.3‬أﺑﻮ ﻳﻮﻧﺲ وزﻣﻼؤهﺎ‪ .(2007 ،‬آﻤﺎ‬
‫أﻇﻬ ﺮت اﻟﺪراﺳ ﺎت أن اﻟﻔﻠ ﻮرا اﻟﻤﺤﺒ ﺔ ﻟﻠﺤ ﺮارة اﻟﻤﺘﻮﺳ ﻄﺔ )وه ﻲ ﺗﻨﺘﻤ ﻲ إﻟ ﻰ اﻟﻔﻠ ﻮرا اﻟﻄﺒﻴﻌﻴ ﺔ‬
‫ﻟﻠﺤﻠﻴﺐ( ﻣﻦ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﺗﻜﻮن ﺳﺎﺋﺪ ًة ﻓﻲ ﺟﺒﻦ اﻟ ﺸﻴﺪر وذﻟ ﻚ ﺑﺎﻋﺘﺒ ﺎر أن ﻣﻌﻈ ﻢ اﻷﺟﺒ ﺎن‬

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‫ﺼﻨﱠﻊ ﺑﺪءًا ﻣﻦ ﺣﻠﻴﺐ ﻏﻴﺮ ﻣﻌﺎﻣﻞ ﺣﺮارﻳﺎً‪ ،‬إﺿﺎﻓﺔ إﻟﻰ هﺬﻩ اﻟﻔﻠﻮرا اﻟﻤ ﺬآﻮرة ﺁﻧﻔ ًﺎ‬
‫اﻟﺒﻠﺪﻳﺔ اﻟﺘﻘﻠﻴﺪﻳﺔ ُﺗ َ‬
‫ﻗﺪ ﻧﺠﺪ ﺑﻜﺘﻴﺮﻳﺎ ﺗﻨﺘﻤﻲ إﻟﻰ اﻟﺠﻨﺲ ‪.(Fitzimons et al., 1999) Pediococcus‬‬
‫‪ 4.2‬ﺗﺼﻨﻴﻊ اﻟﻠﺒﻦ‪:‬‬
‫اﻟﻠﺒﻦ هﻮ ﻣﻨﺘﺞ ﺣﻠﻴﺒﻲ ﻣﺘﺨﻤﺮ ﻣﺘﻤﺎﺳﻚ اﻟﻘﻮام ﻧﺸﺄت ﺻﻨﺎﻋﺘﻪ ﻓﻲ ﺑﻠﻐﺎرﻳﺎ ﺛﻢ اﻧﺘﺸﺮ إﻧﺘﺎﺟﻪ‬
‫واﺳﺘﻬﻼآﻪ إﻟﻰ ﻣﺨﺘﻠﻒ أﻧﺤﺎء اﻟﻌﺎﻟﻢ‪ ،‬ﺣﻴﺚ ﻳﺨﺘﻠﻒ اﻟﻠﺒﻦ اﻟﻤﻨﺘﺞ ﻣﻦ ﻣﻨﻄﻘﺔ إﻟﻰ أﺧﺮى ﻓﻲ اﻟﻌﺎﻟﻢ‬
‫ﻣﻦ ﺣﻴﺚ اﻟﻠﺰوﺟﺔ واﻟﻨﻜﻬﺔ واﻟﻄﻌﻢ‪ .‬آﻤﺎ أﻧﻪ ﻣﻦ اﻟﻤﻤﻜﻦ اﺳﺘﺨﺪام ﺣﻠﻴﺐ اﻟﺤﻴﻮاﻧﺎت اﻟﻤﺨﺘﻠﻔﺔ‬
‫ﻹﻧﺘﺎج اﻟﻠﺒﻦ رﻏﻢ أن ﺣﻠﻴﺐ اﻷﺑﻘﺎر هﻮ اﻷآﺜﺮ اﺳﺘﺨﺪاﻣًﺎ‪ .‬وﻳﻤﻜﻦ إﻧﺘﺎج اﻟﻠﺒﻦ اﻟﻤﺘﺨﻤﺮ ﻣﻦ اﻟﺤﻠﻴﺐ‬
‫آﺎﻣﻞ اﻟﺪﺳﻢ أو اﻟﺤﻠﻴﺐ ﻣﻨﺰوع اﻟﺪﺳﻢ ﺟﺰﺋﻴﺎ" أو اﻟﺤﻠﻴﺐ اﻟﺨﺎﻟﻲ ﻣﻦ اﻟﺪﺳﻢ‪.‬‬
‫هﻨﺎك ﻋﺪة ﺷﺮوط ﻳﻔﺘﺮض ﺗﻮﻓﺮهﺎ ﻓﻲ اﻟﺤﻠﻴﺐ اﻟﻤﺴﺘﺨﺪم ﻹﻧﺘﺎج اﻟﻠﺒﻦ اﻟﻤﺘﺨﻤﺮ أهﻤﻬﺎ أن‬
‫ﻳﻜﻮن اﻟﻤﺤﺘﻮى اﻟﺒﻜﺘﻴﺮي ﻟﻠﺤﻠﻴﺐ ﻣﺘﺪﻧﻴًﺎ وأن ﻳﺨﻠﻮ اﻟﺤﻠﻴﺐ ﻣﻦ ﺑﻘﺎﻳﺎ اﻟﻤﻀﺎدات اﻟﺤﻴﻮﻳﺔ اﻟﻨﺎﺟﻤﺔ‬
‫ﻋﻦ ﻣﻌﺎﻟﺠﺔ ﺑﻌﺾ اﻟﺤﻴﻮاﻧﺎت اﻟﻤﻨﺘﺠﺔ ﻟﻠﺤﻠﻴﺐ )ﺣﻴﺚ ﻳﺠﺐ ﻋﺪم ﺧﻠﻂ ﺣﻠﻴﺐ اﻟﺤﻴﻮاﻧﺎت اﻟﻤﻌﺎﻟﺠﺔ‬
‫ﺑﺎﻟﻤﻀﺎدات اﻟﺤﻴﻮﻳﺔ ﻣﻊ ﺣﻠﻴﺐ اﻷﺑﻘﺎر اﻟﺴﻠﻴﻤﺔ(‪ ،‬آﻤﺎ ﻳﻔﺘﺮض أن ﺗﻜﻮن اﻷﺑﻘﺎر اﻟﻤﻨﺘﺠﺔ ﻟﻠﺤﻠﻴﺐ‬
‫ﻏﻴﺮ ﻣﺼﺎﺑﺔ ﺑﻤﺮض اﻟﺘﻬﺎب اﻟﻀﺮع‪ ،‬وأن ﻳﻜﻮن اﻟﺤﻠﻴﺐ ﺧﺎﻟﻴًﺎ ﻣﻦ اﻟﻠﺒﺄ )اﻟﺴﺮﺳﻮب( اﻟﺬي ُﻳﻔْﺮَز‬
‫ﺑﻌﺪ وﻻدة أﻧﺜﻰ اﻟﺤﻴﻮان ﻣﺒﺎﺷﺮة‪ ،‬وﻳﻜﻮن ُﻣ َﻌﺪًا ﻟﺘﻐﺬﻳﺔ اﻟﻮﻟﻴﺪ ) ;‪Danielsen and Wind, 2003‬‬

‫‪ .(Cintas et al., 1995‬زد ﻋﻠﻰ ذﻟﻚ‪ ،‬أﻧﻪ ﻳﺘﻮﺟﺐ ﺧﻠﻮ اﻟﺤﻠﻴﺐ اﻟ ُﻤ َﻌﺪ ﻹﻧﺘﺎج اﻟﻠﺒﻦ اﻟﻤﺘﺨﻤﺮ ﻣﻦ‬
‫اﻟﻄﻌﻮم ﻏﻴﺮ اﻟﻤﺮﻏﻮﺑﺔ ﻣﺜﻞ ﻃﻌﻢ اﻟﺘﺰﻧﺦ وآﺬﻟﻚ ﺧﻠﻮﻩ ﻣﻦ أي أﺛﺮ ﻟﻠﻤﻨﻈﻔﺎت اﻟﻜﻴﻤﻴﺎﺋﻴﺔ اﻟﻤﺴﺘﺨﺪﻣﺔ‬
‫ﻓﻲ ﻏﺴﻞ أواﻧﻲ اﻟﺤﻠﻴﺐ‪.‬‬
‫ﻳﻤﻜﻦ إﺿﺎﻓﺔ ﻣﻜﻮﻧﺎت أﺧﺮى ﺧﻼل ﺗﺼﻨﻴﻊ اﻟﻠﺒﻦ اﻟﻤﺘﺨﻤﺮ ﺑﺎﻹﺿﺎﻓﺔ ﻟﻠﺤﻠﻴﺐ واﻟﺒﺎدئ‬
‫اﻟﺒﻜﺘﻴﺮي )اﻟﺮوﺑﺔ( ﻣﺜﻞ إﺿﺎﻓﺔ اﻟﺤﻠﻴﺐ ﻣﻨﺰوع اﻟﺪﺳﻢ اﻟﻤﺮآﺰ أو اﻟﺤﻠﻴﺐ ﺧﺎﻟﻲ اﻟﺪﺳﻢ اﻟﻤﺠﻔﻒ أو‬
‫ﻣﺼﻞ اﻟﺤﻠﻴﺐ أو ﺳﻜﺮ اﻟﻼآﺘﻮز‪ ،‬وﻏﺎﻟﺒًﺎ ﻣﺎ ﺗﻀﺎف ﺗﻠﻚ اﻟﻤﻜﻮﻧﺎت ﻟﺰﻳﺎدة ﻣﺤﺘﻮى اﻟﻤﻮاد اﻟﺼﻠﺒﺔ‬
‫ﺤِﻠﻴﺎت ﻣﺜﻞ ﺳﻜﺮ اﻟﻐﻠﻮآﻮز أو اﻟﺴﻜﺮوز أو‬
‫ﻓﻲ اﻟﻠﺒﻦ‪ .‬وﻣﻦ اﻟﻤﻤﻜﻦ أﻳﻀًﺎ إﺿﺎﻓﺔ ﺑﻌﺾ اﻟ ُﻤ َ‬
‫ﺤِﻠﻴﺎت اﻟﺼﻨﺎﻋﻴﺔ ﻋﺎﻟﻴﺔ اﻟﺤﻼوة آﺎﻷﺳﺒﺎرﺗﺎم‪ ،‬وﻗﺪ ﺗﻀﺎف ﺑﻌﺾ ﻣﺜﺒﺘﺎت اﻟﻘﻮام ﻣﺜﻞ اﻟﺠﻴﻼﺗﻴﻦ‬
‫اﻟ ُﻤ َ‬
‫واﻟﻜﺎرﺑﻮآﺴﻲ ﻣﻴﺘﻴﻞ ﺳﻠﻴﻠﻮز واﻟﻜﺎراﺟﻴﻨﺎن وﻏﻴﺮهﺎ‪ .‬أو ﺗﻀﺎف أﻳﻀﺎ ﻗﻄﻊ ﻣﻦ اﻟﻔﺎآﻬﺔ أواﻟﻨﻜﻬﺎت‬
‫وﻏﻴﺮهﺎ‪.‬‬
‫ﻳﺘﻜﻮن اﻟﺒﺎدئ اﻟﺒﻜﺘﻴﺮي اﻟﻤﺴﺘﺨﺪم ﻓﻲ إﻧﺘﺎج اﻟﻠﺒﻦ ﻋﺎد ًة‪ ،‬ﻣﻦ ﺧﻠﻴﻂ ﻣﻦ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ‬
‫اﻟﻌﺼﻮﻳﺔ وﺗﻠﻚ اﻟﻜﺮوﻳﺔ‪ ،‬آﻤﺎ ﻳﺠﻮز اﺳﺘﺨﺪام إﺣﺪاهﻤﺎ ﻓﻘﻂ ﻹﻧﺘﺎج اﻟﻠﺒﻦ‪ .‬إﻻ أن ﻣﻌﺪل إﻧﺘﺎج‬
‫ﺣﻤﺾ اﻟﻠﺒﻦ ﻳﻜﻮن أآﺒﺮ ﻋﻨﺪ اﺳﺘﺨﺪام اﻟﺮوﺑﺔ اﻟﻤﺤﺘﻮﻳﺔ ﻋﻠﻰ ﻧﻮﻋﻲ اﻟﺒﻜﺘﻴﺮﻳﺎ ﻣﻌًﺎ‪ .‬إذ أﻧﻪ ﻣﻦ‬
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‫اﻟﻤﻌﺮوف أن اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﻜﺮوﻳﺔ ﺗﻨﻤﻮ ﺑﺸﻜﻞ أﺳﺮع وﺗﻨﺘﺞ اﻟﺤﻤﺾ وﺛﺎﻧﻲ أآﺴﻴﺪ اﻟﻜﺮﺑﻮن اﻟﻠﺬﻳْﻦ‬
‫ﻳﺤﻔﺰان ﺑﺪورهﻤﺎ ﻧﻤﻮ وﻧﺸﺎط اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﻌﺼﻮﻳﺔ‪ ،‬وﺗﻨﺘﺞ هﺬﻩ اﻷﺧﻴﺮة ﺑﻌﺾ اﻷﺣﻤﺎض اﻷﻣﻴﻨﻴﺔ‬
‫واﻟﺒﺒﺘﻴﺪات اﻟﺘﻲ ُﺗﺴْ َﺘﺨْ َﺪم ﻻﺣﻘًﺎ ﻣﻦ ﻗﺒﻞ اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﻜﺮوﻳﺔ )‪.(Tserovska et al., 2002‬‬
‫ﻳﻨﺘﺞ ﻋﻦ ﻧﺸﺎط اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﻤﺒﻴﻨﺔ أﻋﻼﻩ ﻗﻮام وﻧﻜﻬﺔ اﻟﻠﺒﻦ اﻟﻤﺴﺘﺤﺒﺔ‪ .‬إذ ﻳﺤﺘﻮي اﻟﻠﺒﻦ اﻟﺮاﺋﺐ‬
‫ﻋﻠﻰ ﻣﺮآﺒﺎت ﻧﺎﺗﺠﺔ ﻋﻦ ﻋﻤﻠﻴﺔ اﻟﺘﺨﻤﺮ ﺗﻤﻨﺤﻪ اﻟﻨﻜﻬﺔ اﻟﻤﻤﻴﺰة‪ ،‬ﻣﺜﻞ ﺣﻤﺾ اﻟﻠﺒﻦ واﻷﺳﻴﺖ أﻟﺪﻳﻬﻴﺪ‬
‫وﺣﻤﺾ اﻷﺳﻴﺘﻴﻚ واﻟﺪي أﺳﺘﻴﻞ‪.‬‬
‫ﻳﺘﻢ ﺗﺼﻨﻴﻊ اﻟﻠﺒﻦ اﻟﻤﺘﺨﻤﺮ )اﻟﺮاﺋﺐ( ﺑﺘﺼﻔﻴﺔ اﻟﺤﻠﻴﺐ ﺛﻢ ﺗﻌﺪﻳﻞ ﻣﺤﺘﻮاﻩ ﻣﻦ اﻟﺪهﻮن إذا ﻟﺰم‬
‫اﻷﻣﺮ‪ .‬ﺑﻌﺪ ذﻟﻚ ُﻳ َﺒﺴْ َﺘﺮ اﻟﺤﻠﻴﺐ ﺑﺘﺴﺨﻴﻨﻪ ﻟﻤﺪة ‪ 30‬دﻗﻴﻘﺔ إﻟﻰ اﻟﺪرﺟﺔ ‪˚85‬م أو ﻟﻤﺪة ‪ 10‬دﻗﺎﺋﻖ إﻟﻰ‬
‫اﻟﺪرﺟﺔ ‪˚95‬م‪ .‬ﺣﻴﺚ ﻳﻼﺣﻆ أن اﻟﻤﻌﺎﻣﻠﺔ اﻟﺤﺮارﻳﺔ ﻟﻠﺤﻠﻴﺐ اﻟﻤﻌﺪ ﻹﻧﺘﺎج اﻟﻠﺒﻦ اﻟﻤﺘﺨﻤﺮ ﺗﻜﻮن‬
‫ﺑﺎﺳﺘﺨﺪام ﺣﺮارة أدﻧﻰ وﻟﻤﺪة أﻗﺼﺮ ﻣﻨﻬﺎ ﻓﻲ ﺣﺎﻟﺔ إﻧﺘﺎج اﻟﺤﻠﻴﺐ اﻟﻤﺒﺴﺘﺮ وذﻟﻚ ﻟﻠﻮﺻﻮل إﻟﻰ ﺑﻴﺌﺔ‬
‫ﻣﻨﺎﺳﺒﺔ ﻟﻌﻤﻞ اﻟﺒﺎدئ اﻟﺒﻜﺘﻴﺮي‪ ،‬وﻹﻋﺎدة ﺗﺸﻜﻴﻞ وﺗﺮﺳﻴﺐ ﺑﺮوﺗﻴﻨﺎت ﻣﺼﻞ اﻟﺤﻠﻴﺐ ﻣﻤﺎ ﻳﺴﺎهﻢ ﻓﻲ‬
‫إﻋﻄﺎء اﻟﻠﺰوﺟﺔ واﻟﻘﻮام اﻟﻤﻨﺎﺳﺒﻴﻦ ﻟﻠﺒﻦ اﻟﻤﺘﺨﺜﺮ‪ .‬آﻤﺎ ﺗﺘﺮاﻓﻖ ﻋﻤﻠﻴﺔ اﻟﺒﺴﺘﺮة ﻣﻊ ﻣﺠﺎﻧﺴﺔ اﻟﺤﻠﻴﺐ‪،‬‬
‫ﺣﻴﺚ ﺗﻬﺪف هﺬﻩ اﻟﻌﻤﻠﻴﺔ إﻟﻰ ﻣﻨﻊ ﺗﺸﻜﻞ ﻃﺒﻘﺔ آﺮﻳﻤﺔ )ﻗﺸﺪة( ﻋﻠﻰ ﺳﻄﺢ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ )اﻟﻤﺘﺨﻤﺮ(‬
‫ﺧﻼل ﻋﻤﻠﻴﺔ اﻟﺤﻀﻦ وﺧﻼل ﺣﻔﻆ اﻟﻠﺒﻦ داﺧﻞ اﻟﺜﻼﺟﺔ‪ ،‬وآﺬﻟﻚ ﻟﺘﺤﺴﻴﻦ ﺛﺒﺎﺗﻴﺔ وﻗﻮام اﻟﻠﺒﻦ‪ .‬ﺛﻢ ﻳﺘﻢ‬
‫ﺗﺒﺮﻳﺪ اﻟﺤﻠﻴﺐ إﻟﻰ درﺟﺔ اﻟﺤﺮارة اﻟﻤﺜﻠﻰ ﻟﻌﻤﻞ اﻟﺒﺎدئ اﻟﺒﻜﺘﻴﺮي وهﻲ ‪˚43‬م وﻋﻨﺪهﺎ ُﻳﻀﺎف‬
‫ﺐ ﻣﺘﺴﺎوﻳﺔ ﻣﻦ ﺑﻜﺘﻴﺮﻳﺎ‬
‫ﻞ ﻣﻦ ﻧﺸﺎﻃﻪ وآﻔﺎءﺗﻪ ﺑﻤﺎ ﻳﺤﺘﻮﻳﻪ ﻣﻦ ﻧﺴ ٍ‬
‫اﻟﺒﺎدئ اﻟﺒﻜﺘﻴﺮي اﻟﺬي ﻳﺘﻢ ﺗﻘﻴﻴﻢ آ ٍ‬
‫ﺣﻤﺾ اﻟﻠﺒﻦ اﻟﻜﺮوﻳﺔ وﺗﻠﻚ اﻟﻌﺼﻮﻳﺔ‪ .‬ﺑﻌﺪ إﺿﺎﻓﺔ اﻟﺒﺎدئ اﻟﺒﻜﺘﻴﺮي ﻳﺘﻢ ﺣﻀﻦ اﻟﺤﻠﻴﺐ ﻟﻤﺪة‬
‫ﺗﺘﺮاوح ﻣﺎﺑﻴﻦ ‪ 3‬و‪ 5‬ﺳﺎﻋﺎت ﺑﺪون ﺗﺤﺮﻳﻚ ﻟﺘﺘﻢ ﻋﻤﻠﻴﺔ اﻟﺘﺨﻤﺮ‪ .‬ﺑﻌﺪهﺎ ُﻳ َﺒ ﱠﺮد اﻟﻠﺒﻦ اﻟﻤﺘﺨﻤﺮ إﻟﻰ‬
‫ﻄ ﱠﻌﻤًﺎ ) ‪Revol‬‬
‫درﺟﺔ اﻟﺤﺮارة ‪˚5‬م‪ ،‬وهﻨﺎ ﻗﺪ ﺗﻀﺎف اﻟﻔﻮاآﻪ واﻟﻨﻜﻬﺎت إذا آﺎن اﻟﻠﺒﻦ ُﻣ َﻨ ﱠﻜﻬًﺎ أو ُﻣ َ‬
‫‪.(and Herbin, 1999‬‬

‫وﻳﻤﻜﻨﻨﺎ أن ﻧﻀﻴﻒ هﻨﺎ أهﻤﻴﺔ دﺧﻮل اﻟﻠﺒﻦ ﻓ ﻲ ﺻ ﻨﺎﻋﺔ اﻟﻤﺜﻠﺠ ﺎت ﻟﻠﺤ ﺼﻮل ﻋﻠ ﻰ ﺁﻳ ﺲ آ ﺮﻳﻢ‬
‫ل ﻣﻦ اﻟﻌﻮاﻣﻞ اﻟﻤﻤﺮﺿﺔ ﻣﺜﻞ اﻟـ ‪ .Enterococcus‬إﺿﺎﻓ ًﺔ إﻟﻰ ذﻟﻚ‪ ،‬ﻓﺈن اﺳﺘﺨﺪام اﻟﻠﺒﻦ ﻓﻲ ه ﺬﻩ‬
‫ﺧﺎ ٍ‬
‫اﻟ ﺼﻨﺎﻋﺔ ﻳﻘﻠ ﻞ ﻣ ﻦ اﺣﺘﻤ ﺎﻻت ﺣ ﺪوث اﻟﺘ ﺴﻤﻢ اﻟ ﺬي ﺗ ﺴﺒﺒﻪ اﻟﺘﻮآ ﺴﻴﻨﺎت أو اﻟﺒﻜﺘﻴﺮﻳ ﺎ اﻟﻤﻤﺮﺿ ﺔ‬
‫اﻟﻤﻮﺟﻮدة ﻓﻲ اﻟﻤﻨﺘﺞ وذﻟﻚ ﺑﻔﻀﻞ ﺗﻔﻮق ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﻋﻠﻰ ﺗﻠﻚ اﻟﻌﻮاﻣﻞ اﻹﻣﺮاﺿﻴﺔ‪.‬‬
‫وﻧﻈ ﺮًا ﻷهﻤﻴ ﺔ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ آﺒﺎدﺋ ﺎت ُﺗ ﺴْ َﺘﺨْﺪَم ﻓ ﻲ إﻧﺘ ﺎج اﻷﻟﺒ ﺎن اﻟﻤﺘﺨﻤ ﺮة‪ ،‬وﻧ ﺪرة‬
‫اﻟﺪراﺳﺎت اﻟﺘﺼﻨﻴﻔﻴﺔ ﻟﻬﺬﻩ اﻟﺒﻜﺘﻴﺮﻳﺎ وﻻﺳﻴﻤﺎ ﺗﻠﻚ اﻟﺘﻲ ﺗﺪرس اﻟﻤﻨﺘﺠﺎت اﻟﻤﺤﻠﻴﺔ ﻣﻦ ﻧﺎﺣﻴ ﺔ‪ ،‬وﻋﻠ ﻰ‬
‫اﻋﺘﺒ ﺎر أن ﻣﻌﻈ ﻢ اﻟﻤﻨﺘﺠ ﺎت اﻟﻠﺒﻨﻴ ﺔ اﻟﻤﺤﻠﻴ ﺔ ُﺗﻨْ ﺘَﺞ ﺑﻄﺮﻳﻘ ﺔ ﺗﻘﻠﻴﺪﻳ ﺔ ﺑﺎﺳ ﺘﻌﻤﺎل ﺣﻠﻴ ﺐ ﻏﻴ ﺮ ﻣﻌﺎﻣ ﻞ‬
‫‪14‬‬
‫ﺣﺮارﻳ ًﺎ ﻣ ﻦ ﻧﺎﺣﻴ ﺔ أﺧ ﺮى‪ ،‬ﻓﻘ ﺪ ه ﺪﻓﺖ دراﺳ ﺘﻨﺎ إﻟ ﻰ ﻋ ﺰل وﺗﻨﻤ ﻴﻂ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ وذﻟ ﻚ‬
‫ﺑﺎﺳﺘﺨﺪام ﻃﺮاﺋﻖ اﻟﺘﺼﻨﻴﻒ اﻟﺤﺪﻳﺜﺔ )ﺗﻄﺒﻴﻖ ﺗﻘﻨﻴﺔ اﻟـ ‪ PCR‬وﻣﻄﻴﺎﻓﻴﺔ ﺗﺤﻮﻳﻞ ﻓﻮرﻳﻴﻪ ﻟﻸﺷﻌﺔ ﺗﺤﺖ‬
‫اﻟﺤﻤﺮاء ‪ (FT-IR‬ﺗﻤﻬﻴﺪًا ﻻﺳﺘﺨﺪام اﻷﻧﻮاع اﻷﻓﻀﻞ آﺒﺎدﺋﺎت ﻓ ﻲ ﺗ ﺼﻨﻴﻊ ﺑﻌ ﺾ ﻣﻨﺘﺠ ﺎت اﻟﺤﻠﻴ ﺐ‬
‫اﻟﻤﺤﻠﻴﺔ ﺑﻄﺮﻳﻘﺔ ﺻﺤﻴﺔ وﻣﺘﻮاﻓﻘﺔ ﻣﻊ ذوق اﻟﻤﺴﺘﻬﻠﻚ اﻟﻤﺤﻠﻲ‪.‬‬

‫‪15‬‬
 ‫‪ .2‬اﻟﻄﺮاﺋﻖ‬
‫‪ 1.2‬اﻻﻋﺘﻴﺎن‬
‫ﺼ ﱠﻨﻌﺔ ﺑﺎﻟﻄﺮﻳﻘﺔ اﻟﺘﻘﻠﻴﺪﻳ ﺔ‬
‫ﺗﻢ إﺣﻀﺎر ‪ 96‬ﻋﻴﻨﺔ ﺟﺒﻨﺔ ﺑﻠﺪﻳﺔ وﻟﺒﻦ راﺋﺐ )ﻣﻦ اﻷﺑﻘﺎر واﻷﻏﻨﺎم( ُﻣ َ‬
‫ﻣﻦ ﻣﻨﺎﻃﻖ ﻣﺨﺘﻠﻔﺔ ﻣﻦ اﻟﻘﻄﺮ اﻟﻌﺮﺑﻲ اﻟﺴﻮري‪ ،‬أﺛﻨﺎء ﻣﺪة اﻟﺪراﺳﺔ‪.‬‬
‫ﻀﺮَت ﺳﻠﺴﻠﺔ ﻣ ﻦ اﻟﺘﻤﺪﻳ ﺪات اﻋﺘﺒ ﺎرًا ﻣ ﻦ‬
‫ﺣ ﱢ‬
‫ﺗﻢ ﻋﺰل ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﻣﻦ اﻟﻌﻴﻨﺎت‪ ،‬ﺣﻴﺚ ُ‬
‫اﻟﻤﺤﻠ ﻮل اﻷم ) ذي اﻟﺘﺮآﻴ ﺰ ‪ 1‬غ‪/‬ﻣ ﻞ( ﻟﻤﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟﻤﺘﺨﻤ ﺮة اﻟﻤ ﺮاد ﻋ ﺰل ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ‬
‫اﻟﻠﺒﻦ ﻣﻨﻬﺎ وﻣﻦ ﺛﻢ ُز ِرﻋَﺖ هﺬﻩ اﻟﺘﻤﺪﻳﺪات ﻋﻠﻰ اﻟﻮﺳﻂ اﻟﻤﻼﺋﻢ‪.‬‬

‫‪ 2.2‬اﻟﻌﺰل اﻟﺒﻜﺘﻴﺮي‬
‫ﻀﺮَت ﺑﻴﺌﺘﻲْ اﻻﺳﺘﻨﺒﺎت ‪ MRS‬و‪ M17‬ﻟﻌﺰل اﻟﻌﺼﻴﺎت )‪ (Lactobacillus‬واﻟﻤﻜﻮرات‬
‫ﺣ ﱢ‬
‫ُ‬
‫ْ‬
‫‪37‬‬ ‫ﻀ َﻨﺖ ﻣﺠﻤﻮﻋﺔ ﻣﻦ أﻃﺒﺎق اﻟﺒﺘﺮي ﻓﻲ اﻟﺪرﺟﺔ‬
‫ﺣ ِ‬
‫)‪ (Streptococcus‬اﻟﻠﺒﻨﻴﺔ ﻋﻠﻰ اﻟﺘﻮاﻟﻲ‪ .‬ﺛﻢ ُ‬
‫ﻀ َﻨﺖ اﻟﻤﺠﻤﻮﻋﺔ اﻷﺧﺮى‬
‫ﺣ ِ‬
‫م ﻟﻤﺪة ‪ 48‬ﺳﺎﻋﺔ ﻓﻲ ﻇﺮوف ﻻهﻮاﺋﻴﺔ ﻟﻌﺰل اﻟﻌﺼﻴﺎت اﻟﻠﺒﻨﻴﺔ‪ .‬ﺑﻴﻨﻤﺎ ُ‬
‫‪ 31‬م ﻟﻤﺪة ‪ 48‬ﺳﺎﻋﺔ‪ ،‬وذﻟﻚ ﻣﻦ أﺟﻞ اﻟﺘﻤﻴﻴﺰ ﺑﻴﻦ اﻷﻧﻮاع اﻟﻤﺤﺒﺔ‬
‫‪ 42‬م و ْ‬
‫ﻓﻲ درﺟﺘﻲْ اﻟﺤﺮارة ْ‬
‫ﻟﻠﺤﺮارة اﻟﻤﺮﺗﻔﻌﺔ واﻟﻤﺤﺒﺔ ﻟﻠﺤﺮارة اﻟﻤﺘﻮﺳﻄﺔ ﻣﻦ اﻟﻤﻜﻮرات اﻟﻠﺒﻨﻴﺔ‪.‬‬

‫‪ 3.2‬اﻟﻔﺤﺺ اﻟﻤﺠﻬﺮي‪:‬‬
‫ﺟﺮى اﻟﻔﺤﺺ اﻟﻤﺠﻬﺮي ﺑﺎﺗﺒﺎع اﻟﺨﻄﻮات اﻟﺘﺎﻟﻴﺔ‪:‬‬
‫‪ ‬وﺿﻊ اﻟ ُﻤ َﻌﱠﻠﻖ اﻟﺒﻜﺘﻴﺮي ﻋﻠﻰ ﺻﻔﻴﺤﺔ زﺟﺎﺟﻴﺔ‪.‬‬
‫‪ ‬ﺗﺠﻔﻴﻒ اﻟ ُﻤ َﻌﻠﱠﻖ وﺗﺜﺒﻴﺘﻪ ﺑﻮاﺳﻄﺔ اﻟﻠﻬﺐ‪.‬‬
‫‪ ‬اﻟﺘﻠﻮﻳﻦ ﺑﺎﻟﻔﻴﻮﺷﻴﺴﻴﻦ زﻳﻞ‪ -‬ﻧﻴﻠﺴﻦ ﻣﺪة ‪ 10‬دﻗﺎﺋﻖ‪.‬‬
‫‪ ‬ﻏﺴﻞ اﻟﺼﻔﻴﺤﺔ ﺑﺎﻟﻤﺎء اﻟﺠﺎري‪.‬‬
‫‪ ‬ﺗﻠﻮﻳﻦ اﻟﺼﻔﻴﺤﺔ ﺑﻮاﺳﻄﺔ ﺣﻤﺾ اﻟﺴﻴﺘﺮات ‪ %3‬ﻟﻤﺪة دﻗﻴﻘﺔ‪.‬‬
‫‪ ‬اﻟﻐﺴﻴﻞ ﺑﺎﻟﻤﺎء اﻟﺠﺎري ﻣﺮة ﺛﺎﻧﻴﺔ‪.‬‬
‫‪ ‬إﺿﺎﻓﺔ ﻣﺤﻠﻮل أﺧﻀﺮ ﻣﺎﻟﺸﻴﺖ ‪ %1 Malachite‬ﻟﻤﺪة ‪ 20‬ﺛﺎﻧﻴﺔ‪.‬‬
‫‪ ‬ﺗﺠﻔﻴﻒ اﻟﺼﻔﻴﺤﺔ‪ ،‬وإﺟﺮاء اﻟﻔﺤﺺ اﻟﻤﺠﻬﺮي‪.‬‬

‫‪16‬‬
 ‫‪ 4.2‬اﻻﺧﺘﺒﺎرات اﻟﻜﻴﻤﻴﺎﺋﻴﺔ اﻟﺤﻴﻮﻳﺔ‬
‫ُأﺟْ ِﺮﻳَﺖ اﺧﺘﺒﺎرات اﻷوآﺴﻴﺪاز واﻟﻜﺎﺗﺎﻻز وﺗﺨﻤﻴﺮ اﻟﺴﻜﺎآﺮ )اﻟﺴﻜﺮوز واﻟﻐﻠﻮآﻮز‬
‫واﻟﻼآﺘﻮز( ﺑﻮاﺳﻄﺔ ﺑﻴﺌﺔ ﺣﺎوﻳﺔ ﻋﻠﻰ ﺛﻼﺛﻲ ﺳﻜﺮ اﻟﺤﺪﻳﺪ‪ .‬آﻤﺎ ﺗﻢ اﺧﺘﻴﺎر ﻋﺪد ﻣﻦ اﻟﻌﺰﻻت‬
‫ﻻﺧﺘﺒﺎرهﺎ ﺑﺘﻘﻨﻴﺔ اﻟـ ‪ API 20‬أو اﻟـ ‪:(BioMérieux) API 50‬‬
‫‪ -‬ﺣﻞ اﻟﻤﺴﺘﻌﻤﺮات اﻟﺒﻜﺘﻴﺮﻳﺔ ﺑﻤﺤﻠﻮل ﻣﻠﺤﻲ ‪.%0.85‬‬
‫‪ -‬ﻣﻞء ﺣﻔﺮ ﻗﺎﻋﺪة ﺻﻔﻴﺤﺔ اﻟـ ‪ API‬ﺑﺎﻟﻤﺎء‪.‬‬
‫‪ -‬وﺿﻊ اﻟﻤﺤﻠﻮل اﻟﻤﻠﺤﻲ اﻟﺤﺎوي ﻋﻠﻰ اﻟﺒﻜﺘﻴﺮﻳﺎ ﻓﻲ ﺻﻔﻴﺤﺔ اﻟـ ‪ .API‬ﺛﻢ اﻟﺘﺤﻀﻴﻦ ﻟﻤ ﺪة ‪-24‬‬

‫‪ 48‬ﺳﺎﻋﺔ ﻓﻲ درﺟﺔ اﻟﺤﺮارة اﻟﻤﻨﺎﺳﺒﺔ‪.‬‬

‫ﺿﻌَﺖ اﻟﻜﻮاﺷﻒ اﻟﻤﻨﺎﺳﺒﺔ و ُﻗ ِﺮﺋَﺖ اﻟﻨﺘﻴﺠﺔ ﺑﻤﻘﺎرﻧﺔ اﻟﺼﻔﻴﺤﺔ ﻣﻊ‬
‫‪ -‬ﺑﻌﺪ اﻧﺘﻬﺎء ﻓﺘﺮة اﻟﺤﻀﻦ‪ُ ،‬و ِ‬
‫اﻟﺠﺪاول اﻟﻤﻨﺎﺳﺒﺔ‪.‬‬

‫‪ 5.2‬اﻻﺧﺘﺒﺎرات اﻟﺒﻴﻮﻟﻮﺟﻴﺔ اﻟﺠﺰﻳﺌﻴﺔ‬

‫‪ .1.5.2‬ﻋﺰل اﻟـ ‪:DNA‬‬

‫‪ 37‬م‬
‫ﻀﻨَﺖ ﻓﻲ اﻟﺪرﺟﺔ ْ‬
‫ﺣ ِ‬
‫‪ ‬اﺳْ ُﺘﻨْ ِﺒﺘَﺖ ﻣﺴﺘﻌﻤﺮة ﺑﻜﺘﻴﺮﻳﺔ ﻣﻌﺰوﻟﺔ ﻓﻲ وﺳﻂ اﻻﺳﺘﻨﺒﺎت‪ ،‬و ُ‬
‫ﻟﻤﺪة ﻟﻴﻠﺔ آﺎﻣﻠﺔ‪.‬‬
‫‪ُ ‬أﺧِﺬ ‪ 1.5‬ﻣﻞ ﻣﻦ اﻟﻤﺴﺘﻨﺒﺖ اﻟﺒﻜﺘﻴﺮي وُﺛ ﱢﻔﻞ ﺑﺴﺮﻋﺔ ‪ 9000‬دورة‪/‬د‪ ،‬ﻟﻤﺪة ‪ 20‬ﺛﺎﻧﻴﺔ‪.‬‬
‫ﺿﻊ ‪ 500‬ﻣﻴﻜﺮوﻟﺘﺮ ﻣﻦ ﻣﻮﻗﻲ اﻟـ ‪ TE‬ﻓﻮق اﻟﺮاﺳﺐ اﻟﺒﻜﺘﻴﺮي‪ ،‬ﺛﻢ ُأﺿﻴﻒ ‪50‬‬
‫ُو ِ‬ ‫‪‬‬

‫ﻣﻴﻜﺮوﻟﻴﺘﺮﻣﻦ اﻟﻠﻴﺰوزﻳﻢ )‪ 10‬ﻣﻠﻎ‪/‬ﻣﻞ(‪.‬‬

‫‪ 37‬م‪.‬‬
‫‪ ‬ﺟﺮى رج اﻟﻤﺰﻳﺞ ﺟﻴﺪًا ﺛﻢ ُﺗ ِﺮك ﻟﻤﺪة ﺳﺎﻋﺘﻴْﻦ ﻓﻲ ﺣﻤﺎم ﻣﺎﺋﻲ درﺟﺔ ﺣﺮارﺗﻪ ْ‬
‫‪ 50‬م‪.‬‬
‫‪ُ ‬ﺗ ِﺮك اﻟﻤﺰﻳﺞ ﻟﻴﻠ ًﺔ آﺎﻣﻠ ًﺔ ﻓﻲ اﻟﺪرﺟﺔ ْ‬
‫‪ 37‬م‬
‫ﻦ اﻟﻤﺰﻳﺞ ﻓﻲ اﻟﺪرﺟﺔ ْ‬
‫ﻀَ‬
‫ﺣ ِ‬
‫‪ُ ‬أﺿﻴﻒ ‪ 25‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ اﻟﺒﺮوﺗﻴﻨﺎز ‪ 20) K‬ﻣﻠﻎ‪/‬ﻣﻞ(‪ ،‬و ُ‬
‫ﻟﻤﺪة ﺳﺎﻋﺔ‪.‬‬
‫‪ُ ‬أﺿﻴﻒ ‪ 25‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ‪ (%25) SDS‬ﻣﻊ اﻟﺤﻀﻦ ﻟﻤﺪة ﺳﺎﻋﺔ‪.‬‬
‫‪ُ ‬أﺿﻴﻒ ‪ 200‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ آﻠﻮرﻳﺪ اﻟﺼﻮدﻳﻮم )‪ 5‬ﻣﻮل(‪.‬‬

‫‪17‬‬
 ‫‪ُ ‬أﺿﻴﻒ ‪ 750‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ ﻣﺰﻳﺞ ﻓﻴﻨﻮل‪ :‬آﻠﻮروﻓﻮرم‪ :‬إﻳﺰوأﻣﻴﻞ ) ‪ (1 :24: 25‬ﻣﻊ‬
‫اﻟﻤﺰج اﻟﺠﻴﺪ‪.‬‬
‫‪ُ ‬ﺛﻔﱢﻞ اﻟﻤﺰﻳﺞ ﺑﺴﺮﻋﺔ ‪ 13000‬دورة‪/‬د‪ ،‬ﻟﻤﺪة ‪ 6‬دﻗﺎﺋﻖ‪.‬‬
‫ُأﺿﻴﻒ ‪ 450‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ اﻹﻳﺰوﺑﺮﺑﺎﻧﻮل إﻟﻰ اﻟﻄﺎﻓﻲ و ُﺛﻔﱢﻞ اﻟﻤﺰﻳﺞ ﺑﺴﺮﻋﺔ ‪13000‬‬ ‫‪‬‬

‫دورة‪/‬د ﻟﻤﺪة ‪ 45‬دﻗﻴﻘﺔ‪.‬‬

‫‪ُ ‬أﺿﻴﻒ ‪ 1‬ﻣﻞ ﻣﻦ اﻹﻳﺘﺎﻧﻮل ‪ %70‬إﻟﻰ اﻟﺮاﺳﺐ‪.‬‬
‫ﺟﻔﱢﻒ اﻟﺮاﺳﺐ‬
‫‪ُ ‬ﺛﻔﱢﻞ اﻟﻤﺰﻳﺞ ﺑﺴﺮﻋﺔ ‪ 13000‬دورة‪/‬د ﻟﻤﺪة ‪ 5‬دﻗﺎﺋﻖ ﻓﻲ اﻟﺪرﺟﺔ ‪ ْ4‬م‪ ،‬ﺛﻢ ُ‬
‫ﺑﺎﺳﺘﺨﺪام ﺟﻬﺎز ﺗﺮآﻴﺰ اﻟﺤﻤﻮض اﻟﻨﻮوﻳﺔ )إﻳﺒﻨﺪورف أﻟﻤﺎﻧﻴﺎ(‪.‬‬
‫‪ 20‬م( ﻟﺤﻴﻦ‬
‫ﺣﻔِﻆ اﻟﻤﺰﻳﺞ ﻓﻲ اﻟﻤﺠﻤﺪة )‪ْ -‬‬
‫‪ُ ‬أﺿﻴﻒ ‪ 20‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ ﻣﻮﻗﻲ اﻟـ ‪ TE‬و ُ‬
‫اﻻﺳﺘﺨﺪام‪.‬‬
‫‪ .2.5.2‬ﺗﻘﻨﻴﺔ اﻟـ ‪:PCR‬‬
‫ﺟﺮى ﺗﺤﻀﻴﺮ ﻣ ﺰﻳﺞ ﺣﺠﻤ ﻪ اﻟﻜﻠ ﻲ ‪ 25‬ﻣﻴﻜﺮوﻟﻴﺘ ﺮ ﻳﺤﺘ ﻮي ﻋﻠ ﻰ‪ 2 :‬ﻣﻴﻜﺮوﻟﻴﺘ ﺮ ﻣ ﻦ ﻣﻌﻠ ﻖ اﻟ ـ‬
‫‪ 500) DNA‬ﻧﺎﻧﻮ ﻏﺮام(‪ ،‬ﺑﺎﻹﺿﺎﻓﺔ إﻟ ﻰ ‪ 1.5‬ﻣﻴﻜﺮوﻟﻴﺘ ﺮ ﻣ ﻦ ﻣ ﻮﻗﻲ اﻟﺘﻔﺎﻋ ﻞ‪ ،‬و ‪ 3‬ﻣﻴﻜﺮوﻟﻴﺘ ﺮ ﻣ ﻦ‬
‫آﻠﻮرﻳﺪ اﻟﻤﻨﻐﻨﺰﻳﻮم )‪ 25‬ﻧﺎﻧﻮ ﻣﻮل(‪ ،‬و‪ 0.5‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ ‪ ،(20 mM) dNTPs‬و‪ 1‬ﻣﻴﻜﺮوﻟﻴﺘ ﺮ ﻣ ﻦ‬
‫ﺮ ﱢﺋﺴﺎت اﻟﺘﻲ ﺗﻨﺎﺳﺐ أﺟﻨﺎس اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﻤﺪروﺳﺔ )اﻟﺠﺪول ‪ (1‬و‪ 1‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ أﻧﺰﻳﻢ اﻟـ ‪DNA‬‬
‫اﻟ ُﻤ َ‬
‫‪) polymerase‬هﻴﺌﺔ اﻟﻄﺎﻗﺔ اﻟﺬرﻳﺔ اﻟﺴﻮرﻳﺔ‪ ،‬ﻗﺴﻢ اﻟﺘﻘﺎﻧﺔ(‪ ،‬ﺛﻢ ُأآْ ِﻤﻞ اﻟﺤﺠﻢ ﺑﺎﺳﺘﺨﺪام اﻟﻤﺎء اﻟﻤﻘﻄ ﺮ‪.‬‬
‫ﺗﻢ اﺧﺘﻴﺎر ﻣﻮرﺛﺔ ‪ LacZ‬ﻟﻠﺘﻤﻴﻴﺰ ﺑﻴﻦ اﻟﻨﻮﻋﻴﻦ‪:‬‬
‫‪ Streptococcus thermophilus‬و‪.Enterococcus feacalis‬‬
‫اﻟﺠﺪول )‪ .(1‬اﻟﻤﺮﺋﺴﺎت اﻟﻤﺴﺘﺨﺪﻣﺔ ﻓﻲ اﻟﺪراﺳﺔ‬
‫ﺗﺘﺎﻟﻲ اﻟﻤﺮﺋﺴﺔ‬ ‫اﻟﻤﻮرﺛﺔ‬ ‫اﻟﻨﻮع اﻟﺒﻜﺘﻴﺮي‬ ‫اﻟﺘﺴﻠﺴﻞ‬
‫'‪5'- ATACGTTCCCGGGCCTTGTA -3‬‬ ‫‪16s RNA‬‬ ‫‪Lactobacillus spp.‬‬ ‫‪1‬‬
‫'‪5'- GAACCATCGACCTCACGCTT -3‬‬
‫'‪5'- AACGGGTGAGTAACGCGTGG -3‬‬ ‫‪16s RNA‬‬ ‫‪Lactococcus lactis‬‬ ‫‪2‬‬
‫'‪5'- CCACGCTTTCGAGCCTCAGT -3‬‬
‫'‪5'- AGACAAGCAAGTCTCGCGGA -3‬‬ ‫‪16s RNA‬‬ ‫‪Lactococcus bulgaricus‬‬ ‫‪3‬‬
‫'‪5'- CGCCACTGGTGTTCTTCCA -3‬‬
‫'‪5'- GTGTTGCATGGTGTCG -3‬‬ ‫‪LacZ‬‬ ‫‪Streptococcus‬‬ ‫‪4‬‬
‫'‪5'- CCACCACCAGAGTTACCCAT -3‬‬ ‫‪thermophilus‬‬
‫'‪5'-ATTAATCGGTTGCCTGGACG-3‬‬ ‫‪LacZ‬‬ ‫‪Enterococcus feacalis‬‬ ‫‪5‬‬
‫'‪5'-GCCAATTTCCAGGAACAGTG-3‬‬

‫‪18‬‬
‫ا ُﺗ ِﺒﻌَﺖ اﻟﻤﺮاﺣﻞ اﻟﺘﺎﻟﻴﺔ ﻓﻲ ﺟﻬﺎز اﻟـ ‪ :PCR‬ﻣﺮﺣﻠﺔ اﻟﺘﻤﺴﺦ ‪ْ 95‬م ﻟﻤﺪة ‪ 1‬دﻗﻴﻘﺔ‪ ،‬ﻣﺮﺣﻠﺔ‬
‫اﻻرﺗﺒﺎط ‪ْ 50‬م ﻟﻤﺪة ‪ 1‬دﻗﻴﻘﺔ‪ ،‬ﻣﺮﺣﻠﺔ اﻻﺳﺘﻄﺎﻟﺔ ‪ْ 72‬م ﻟﻤﺪة ‪ 1‬دﻗﻴﻘﺔ‪ ،‬آﻤﺎ ﺗﻀﻤﻦ اﻟﺒﺮﻧﺎﻣﺞ اﻟﺤﻀﻦ‬
‫ﺿ َﻌﺖ اﻷﻧﺎﺑﻴﺐ ﻓﻲ اﻟﺪرﺟﺔ ‪ْ 72‬م ﻟﻤﺪة ‪ 10‬دﻗﺎﺋﻖ‬
‫‪ 72‬م ﻟﻤﺪة ‪ 5‬دﻗﺎﺋﻖ‪ ،‬ﺑﻌﺪ اﻻﻧﺘﻬﺎء ُو ِ‬
‫ﻓﻲ اﻟﺪرﺟﺔ ْ‬
‫آﻤﺮﺣﻠﺔ اﺳﺘﻄﺎﻟﺔ ﻧﻬﺎﺋﻴﺔ‪ .‬وﻓﻲ اﻟﻨﻬﺎﻳﺔ ﺗﻢ اﻟﺘﺒﺮﻳﺪ ﻓﻲ اﻟﺪرﺟﺔ ‪ْ 4‬م ﺑﻌﺪ اﻟﺪورة اﻷﺧﻴﺮة‪ ،‬وﻗﺪ ﺗﻜ ّﻮن‬
‫اﻟﺒﺮﻧﺎﻣﺞ ﻣﻦ ‪ 35‬دورة‪.‬‬
‫‪ .3.5.2‬اﻟﺮﺣﻼن اﻟﻜﻬﺮﺑﺎﺋﻲ‪:‬‬
‫‪ ‬ﺟﺮى وزن ‪ 1‬غ ﻣﻦ اﻵﻏﺎروز وﺗﻢ ﺣﻠﻪ ﻓﻲ ‪ 100‬ﻣﻞ ﻣﻦ اﻟـ ‪ ،TAE‬ﺑﺤﻴﺚ ﺗﻜﻮن درﺟﺔ اﻟـ‬
‫‪.7 = pH‬‬
‫‪ ‬ﺟﺮت إذاﺑﺔ اﻵﻏﺎروز ﻓﻲ اﻟﻤﻴﻜﺮووﻳﻒ ﺣﺘﻰ ﺗﻤﺎم اﻟﺬوﺑﺎن‪.‬‬
‫‪ُ ‬أﺿﻴﻒ إﻟﻰ اﻟﻬﻼﻣﺔ اﻟﺴﺎﺋﻠﺔ ‪ 3‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ اﻹﻳﺘﻴﺪﻳﻮم ﺑﺮوﻣﻴﺪ ﻹﻇﻬﺎر ﻋﺼﺎﺋﺐ‬
‫اﻟﺘﺮﺣﻴﻞ وذﻟﻚ ﺑﺤﺬر ﺷﺪﻳﺪ‪.‬‬
‫ﺻﺒﱠﺖ اﻟﻬﻼﻣﺔ‪ ،‬ﻣﻊ ﻣﺮاﻋﺎة ﻋﺪم ﺗﺸﻜﻞ ﻓﻘﺎﻋﺎت‬
‫ﺿﻌَﺖ أﻣﺸﺎط اﻟﺮﺣﻼن وﻣﻦ ﺛﻢ ُ‬
‫‪ُ ‬و ِ‬
‫واﻟﺤﻔﺎظ ﻋﻠﻰ اﺳﺘﻘﺎﻣﺔ اﻟﻬﻼﻣﺔ ﺑﺸﻜﻞ ﺗﺎم‪ ،‬ﺛﻢ ُﺗ ِﺮآَﺖ ﻟﺘﺠﻒ‪.‬‬
‫‪ُ ‬ﻧ ِﻘﻠَﺖ اﻟﻬﻼﻣﺔ ﺑﻌﺪ ﺗﺤﺮﻳﺮهﺎ إﻟﻰ وﻋﺎء اﻟﺮﺣﻼن‪ ،‬وُأﺿﻴﻒ ﻣﻮﻗﻲ اﻟﺮﺣﻼن ﺑﺸﻜﻞ ﻳﻐﻤﺮ‬
‫اﻵﺑﺎر اﻟﻤﺘﺸﻜﻠﺔ‪.‬‬
‫ﻀﺮَت ﻋﻴﻨﺎت اﻟـ ‪ DNA‬اﻟ ُﻤﺮاد ﺗﺮﺣﻴﻠﻬﺎ ﺑﺤﻴﺚ آﺎن ﺣﺠﻤﻬﺎ اﻟﻨﻬﺎﺋﻲ ‪ 15‬ﻣﻴﻜﺮوﻟﻴﺘﺮ‪.‬‬
‫ﺣ ﱢ‬
‫‪ُ ‬‬
‫ﺿﻊ اﻟﻮاﺳﻢ اﻟﺠﺰﻳﺌﻲ ‪ 100) Marker‬أﺳﺎس ﺁوزﺗﻲ( واﻟﻌﻴﻨﺎت ﻓﻲ اﻵﺑﺎر‪.‬‬
‫‪ُ ‬و ِ‬
‫ﺟ ﱢﻬﺰَت اﻹﻟﻜﺘﺮودات ﺑﺤﻴﺚ ﺗﻢ ﺗﻤﺮﻳﺮ اﻟﺘﻴﺎر اﻟﻜﻬﺮﺑﺎﺋﻲ ﻣﻦ‬
‫ﻄﻲ ﺟﻬﺎز اﻟﺮﺣﻼن‪ ،‬و ُ‬
‫ﻏﱢ‬‫‪ُ ‬‬
‫ﺷﻐﱢﻞ اﻟﺠﻬﺎز ﺑﺸﺪة ‪ 60‬أﻣﺒﻴﺮ ﻟﻤﺪة ﺳﺎﻋﺘﻴْﻦ ﺗﻘﺮﻳﺒًﺎ‪.‬‬
‫اﻟﻘﻄﺐ اﻟﺴﺎﻟﺐ إﻟﻰ اﻟﻘﻄﺐ اﻟﻤﻮﺟﺐ‪ُ .‬‬
‫ﺤﺼَﺖ اﻟﻬﻼﻣﺔ ﺑﻮاﺳﻄﺔ ﺟﻬﺎز إﻇﻬﺎر اﻟﻌﺼﺎﺋﺐ )‪.(gel documentation‬‬
‫‪ُ ‬ﻓ ِ‬

‫‪ 6.2‬ﺗﻘﻨﻴﺔ اﻟـ ‪FT-IR‬‬

‫ﻀﻨَﺖ ﻓ ﻲ اﻟﺪرﺟ ﺔ ْ‬
‫‪ 30‬م‪ .‬ﺗ ﻢ اﺧﺘﻴ ﺎر‬ ‫ﺣ ِ‬
‫ﺟ ﺮى اﺳ ﺘﻨﺒﺎت اﻟﺒﻜﺘﻴﺮﻳ ﺎ ﻋﻠ ﻰ أوﺳ ﺎط ﺻ ﻠﺒﺔ ﺛ ﻢ ُ‬
‫ﻣﺴﺘﻌﻤﺮات ﻣﻔﺮدة ﻧﻘﻴﺔ ﻟﻠﺘﻠﻘﻴﺢ )ﻣﻦ اﻟﻀﺮوري أن ﻳﻜﻮن اﻟﻤﺴﺘﻨﺒﺖ ﺻﺎﻓﻴًﺎ(‪.‬‬
‫ﺗﻢ ﺗﺤﻀﻴﺮ اﻟﻌﻴﻨﺔ ﺣﺴﺐ اﻟﺨﻄﻮات اﻵﺗﻴﺔ‪:‬‬
‫ﺧﺬَت ﻣﺎدة اﻟﻌﻴﻨﺔ ﻣﺒﺎﺷﺮة ﻣﻦ ﻣﺮآﺰ اﻻﺳﺘﻨﺒﺎت ﺑﺎﺳﺘﻌﻤﺎل ﻏﺎﻧﺔ ﻣﻦ اﻟﺒﻼﺗﻴﻨﻴﻮم ﻗﻄﺮهﺎ‬
‫‪ُ .1‬أ ِ‬
‫‪ 1‬ﻣﻠﻢ‪ .‬ﺗﻌﺘﻤﺪ آﻤﻴﺔ اﻟﻌﻴﻨ ﺔ اﻟ ﻀﺮورﻳﺔ ﻋﻠ ﻰ اﻟﻜ ﺎﺋﻦ و"اﺗ ﺴﺎق" ﻧﻤ ﻮ ﻣ ﺴﺘﻌﻤﺮاﺗﻪ‪ ،‬ﻟﻜ ﻦ‬

‫‪19‬‬
‫اﻟﺸﻜﻞ اﻟﻨﻤﻮذﺟﻲ ﻳﺘﻤﺜﻞ ﺑﺄن ﺗﻤﻸ اﻟﻌﻴﻨﺔ ﺣﻠﻘ ًﺔ آﺎﻣﻠ ًﺔ ﻟﻐﺎﻧ ٍﺔ واﺣﺪ ٍة‪ .‬ﻳﺠﺐ أن ﺗﻜ ﻮن ﻣ ﺎدة‬
‫اﻟﻌﻴﻨﺔ ﻣﺄﺧﻮذة ﺣﺼﺮًا ﻣﻦ ﻣﻨﺎﻃﻖ اﻟﻨﻤﻮ اﻟﻤﻨﺪﻣﺞ ﻓﻲ اﻟﻤﺴﺘﻌﻤﺮات‪.‬‬
‫‪ُ .2‬أ ِ‬
‫ﺧ َﺬ ‪ 25‬ﻣﻴﻜﺮوﻟﻴﺘﺮ ﻣﻦ اﻟ ُﻤ َﻌﱠﻠﻖ اﻟﺒﻜﺘﻴﺮي إﻟﻰ ﺑﺌﺮ )ﺻﻔﻴﺤﺔ ‪.(96‬‬
‫ﺟ ﱢﻔﻔَﺖ اﻟﻌﻴﻨﺎت ﻓﻲ اﻟﻔﺮن اﻟﺠﺎف ﻓﻲ درﺟﺔ اﻟﺤﺮارة ْ‪ 40‬م ﻟﻤﺪة ‪ 30‬دﻗﻴﻘﺔ‪.‬‬
‫‪ُ .3‬‬
‫‪ .4‬ﺟ ﺮى ﺗﺤﻠﻴ ﻞ اﻟﻌﻴﻨ ﺎت ﺑﻔ ﻀﻞ اﻟﺒﺮﻧ ﺎﻣﺞ اﻟﺤﺎﺳ ﻮﺑﻲ ‪ OPUS‬وﺗﻤ ﺖ ﻣﻘﺎرﻧﺘﻬ ﺎ ﻣ ﻊ‬
‫اﻟﺴﻼﻻت اﻟﻤﺮﺟﻌﻴﺔ اﻟﺘﻲ ﻳﺤﻮﻳﻬﺎ اﻟﺒﺮﻧﺎﻣﺞ )ﻣﻜﺘﺒﺔ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ(‪.‬‬

‫‪20‬‬
 ‫اﻟﻨﺘﺎﺋﺞ‬ ‫‪.3‬‬
‫‪ 1.3‬ﻧﺘﺎﺋﺞ ﺗﺤﺪﻳﺪ هﻮﻳﺔ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﺑﺘﻘﻨﻴﺔ اﻟـ ‪:PCR‬‬
‫ﺣ ﱢﺪ َدت ﺑﺘﻘﻨﻴ ﺔ اﻟ ـ‬
‫ُد ِرس ﺗﻨﻤﻴﻂ ﻋﺪد ﻣﻦ اﻟﺴﻼﻻت اﻟﺒﻜﺘﻴﺮﻳﺔ ﻣﻦ اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ واﻟﻠﺒﻦ اﻟﺮاﺋ ﺐ ‪ُ -‬‬
‫‪ API 20 Strep‬ﻋﻠﻰ أﻧﻬ ﺎ ﺗﺎﺑﻌ ﺔ ﻟﻠﻨ ﻮع ‪) Streptococcus thermophilus‬أﺑ ﻮ ﻳ ﻮﻧﺲ وزﻣﻼؤه ﺎ(–‬
‫ﺑﺎﺳﺘﺨﺪام ﺗﻘﻨﻴﺔ اﻟـ ‪ .PCR‬وﻗﺪ ﺟﺮى ﻋﺰل ه ﺬﻩ اﻟ ﺴﻼﻻت ﻣ ﻦ ﻣﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟﻤﺘﺨﻤ ﺮة اﻟ ﺴﻮرﻳﺔ‬
‫)ﻟﺒﻦ راﺋﺐ – ﺟﺒﻨﺔ ﺑﻠﺪﻳﺔ( إﺛﺮ ﻧﻤﻮهﺎ ﻋﻠﻰ ﺑﻴﺌﺔ اﻟـ ‪ M17‬ﺑﻌﺪ اﻟﺤﻀﻦ ﻓﻲ درﺟﺔ اﻟﺤﺮارة ‪ْ 42‬م ﻟﻤﺪة‬
‫‪ 48‬ﺳﺎﻋﺔ‪ ،‬ﺣﻴﺚ ﺗ ﻢ اﻻﻋﺘﻤ ﺎد ﻋﻠ ﻰ ﺗ ﻀﺨﻴﻢ ﻣﻮرﺛ ﺔ ‪ LacZ‬وه ﻲ اﻟﻤﻮرﺛ ﺔ اﻟﻤﺤﺎﻓﻈ ﺔ ﻓ ﻲ ﺳ ﻼﻻت‬
‫ﺑﻜﺘﻴﺮﻳﺎ ‪ Streptococcus thermophilus‬واﻟﺘﻲ ﺗﺤﺼﺮ ﺷﺪﻓﺔ ﻃﻮﻟﻬﺎ ﺣﻮاﻟﻲ ‪ 242‬أﺳﺎس ﺁزوﺗﻲ ﻓﻲ‬
‫هﺬﻩ اﻟﺴﻼﻻت‪.‬‬
‫وﻳﻈﻬﺮ اﻟ ﺸﻜﻞ ‪ 1.2) 2‬و ‪ ،(2.2‬ﻓ ﻲ اﻟﻤ ﺴﺎر )‪ (1‬ﻧ ﻮاﺗﺞ اﻟﺘﻔﺎﻋ ﻞ ﻣ ﻊ ﺳ ﻼﻟﺔ ‪S. thermophilus‬‬

‫اﻟﺴﻼﻟﺔ اﻟ ُﻤﺤﻀﺮة ﻣﻦ ﺷﺮآﺔ هﺎﻧﺴﻦ‪ -‬اﻟﺪاﻧﻤﺎرك واﻟﻤﺴﺘﺨﺪﻣﺔ آﺸﺎهﺪ إﻳﺠﺎﺑﻲ ﻟﻬ ﺬا اﻟﻨ ﻮع‪ ،‬وﻳُﻼﺣ ﻆ‬
‫ﻣﻦ اﻟﻤﺴﺎرات ‪ 2‬وﺣﺘﻰ ‪ 4‬ﺗﻮاﻓ ﻖ ﺑ ﺎﻟﺤﺠﻢ اﻟﺠﺰﻳﺌ ﻲ ﻟﻠﻤﻮرﺛ ﺔ ‪ LacZ‬ﻟﻠﻨ ﻮع ﻣ ﻊ ﺳ ﻼﻟﺔ هﺎﻧ ﺴﻦ‪ ،‬ﻣﻤ ﺎ‬
‫ﻳﺸﻴﺮ إﻟﻰ أن هﺬﻩ اﻟﺴﻼﻻت اﻟﺜﻼث هﻲ ﻟﻠﻨﻮع ‪ .S. thermophilus‬وهﺬا ﻣﺎ ﻳﺘﻮاﻓﻖ ﻣ ﻊ ﻧﺘ ﺎﺋﺞ ‪API‬‬

‫‪ 20 Strep‬واﻟﺘﻲ ﺗﺆآﺪ أن اﻟﺴﻼﻻت اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ واﻟﺠﺒﻨ ﺔ اﻟﺒﻠﺪﻳ ﺔ ه ﻲ ﻣ ﻦ اﻟﻨ ﻮع ‪S.‬‬

‫‪.thermophilus‬‬

‫اﻟﺸﻜﻞ )‪ .(1.2‬ﻧﺘﺎﺋﺞ اﻟﺮﺣﻼن اﻟﻜﻬﺮﺑﺎﺋﻲ ﻟﻨﻮاﺗﺞ ﺗﻘﻨﻴﺔ اﻟـ ‪ PCR‬ﺑﺎﺳﺘﺨﺪام ﻣﺮﺋﺴﺎت ﻟﻠﻤﻮرﺛﺔ ‪:LacZ‬‬
‫‪Streptococcus‬‬ ‫‪)Streptcoccus thermophilus:1‬هﺎﻧ ﺴﻦ(اﻟ ﺸﺎهﺪ اﻻﻳﺠ ﺎﺑﻲ‪:4-2 ،‬‬
‫‪ thermophilus‬اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﺠﺒﻨ ﺔ اﻟﺒﻠﺪﻳ ﺔ ﻣ ﻦ ﺑﻌ ﺾ اﻟﻤﻨ ﺎﻃﻖ اﻟ ﺴﻮرﻳﺔ‪Enterococcus :5 ،‬‬
‫‪،fecalis‬اﻟﺸﺎهﺪ اﻟﺴﻠﺒﻲ‪ :Mw .‬اﻟﻮاﺳﻢ اﻟﺠﺰﻳﺌﻲ‪.‬‬
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 ‫اﻟﺸﻜﻞ )‪ .(2.2‬ﻧﺘﺎﺋﺞ اﻟﺮﺣﻼن اﻟﻜﻬﺮﺑﺎﺋﻲ ﻟﻨﻮاﺗﺞ ﺗﻘﻨﻴﺔ اﻟـ ‪ PCR‬ﺑﺎﺳﺘﺨﺪام ﻣﺮﺋﺴﺎت ﻟﻠﻤﻮرﺛﺔ ‪:LacZ‬‬
‫‪)Streptcoccus thermophilus:1 :1‬هﺎﻧ ﺴﻦ(اﻟ ﺸﺎهﺪ اﻻﻳﺠ ﺎﺑﻲ‪Streptococcus :4-2 ،‬‬
‫‪ thermophilus‬اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﻠ ﺒﻦ ﻣ ﻦ ﺑﻌ ﺾ اﻟﻤﻨ ﺎﻃﻖ اﻟ ﺴﻮرﻳﺔ‪Enterococcus :5 ،‬‬
‫‪،fecalis‬اﻟﺸﺎهﺪ اﻟﺴﻠﺒﻲ‪ :Mw .‬اﻟﻮاﺳﻢ اﻟﺠﺰﻳﺌﻲ‪.‬‬

‫إﺿﺎﻓ ًﺔ إﻟﻰ ذﻟﻚ‪ ،‬ﺗﻢ ﻋﺰل ﻋﺪد ﻣﻦ اﻟﺴﻼﻻت اﻟﺒﻜﺘﻴﺮﻳﺔ )ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ وﻣﻦ اﻟﺠﺒﻨﺔ‬
‫اﻟﺒﻠﺪﻳﺔ(‪ ،‬ﺣﻴﺚ ﻇﻬﺮت ﺗﺤﺖ اﻟﻤﺠﻬﺮ ﺑﺄﻧﻬﺎ ﻋﺼﻮﻳﺔ وﻣﻮﺟﺒﺔ اﻟﻐﺮام وذﻟﻚ ﺑﻌﺪ زراﻋﺘﻬﺎ ﻋﻠﻰ ﺑﻴﺌﺔ‬
‫اﻟـ ‪ ROGOSA‬وﺣﻀﻨﻬﺎ ﻟﻤﺪة ‪ 48‬ﺳﺎﻋﺔ ﻓﻲ اﻟﺪرﺟﺔ ْ‪ 37‬م‪ .‬آﺎﻧﺖ ﺟﻤﻴﻊ اﻟﺴﻼﻻت ﺳﺎﻟﺒﺔ‬
‫ﺣ ﱢﺪ َدت هﺬﻩ اﻟﺴﻼﻻت ﺑﺎﺳﺘﺨﺪام ﺗﻘﻨﻴﺔ اﻟـ ‪ API‬ﻋﻠﻰ‬
‫اﻟﻜﺎﺗﺎﻻز‪ ،‬وذﻟﻚ ﺿﻤﻦ ﻇﺮوف ﻻ هﻮاﺋﻴﺔ‪ُ .‬‬
‫أﻧﻬﺎ ﺗﺎﺑﻌﺔ ﻟﻠﺠﻨﺲ ‪.Lactobacillus‬‬
‫وﻣﻦ أﺟﻞ اﻟﺘﻨﻤﻴﻂ ﺑﺎﺳﺘﺨﺪام ﺗﻘﻨﻴﺔ اﻟـ ‪ ،PCR‬اﺳْ ُﺘﺨْ ِﺪ َم ﻋﺪد ﻣﻦ اﻟ ُﻤ َﺮ ﱢﺋﺴﺎت ﻟﻠﻜﺸﻒ ﻋﻦ اﻟﻤﻮرﺛﺔ‬
‫‪ 16sRNA‬وهﻲ ﻣﻮرﺛﺔ ﻣﺤﺎﻓﻈﺔ ﺿﻤﻦ ﺟﻨﺲ ‪) Lactobacillus‬اﻟﺠﺪول ‪ ،(1‬ﺣﻴﺚ أن هﺬﻩ‬
‫اﻟ ُﻤ َﺮﺋﱢﺴﺎت ﺗﺤﺼﺮ ﺷﺪﻓﺔ ﻃﻮﻟﻬﺎ ‪ 425‬أﺳﺎس ﺁزوﺗﻲ‪ .‬آﻤﺎ اﺳْ ُﺘﺨْ ِﺪﻣَﺖ هﺬﻩ اﻟ ُﻤ َﺮ ﱢﺋﺴﺎت ﻟﻠﻜﺸﻒ ﻋﻦ‬
‫اﻟﻤﻮرﺛﺔ ‪ 23SRNA‬وهﻲ أﻳﻀًﺎ ﻣﻮرﺛﺔ ﻣﺤﺎﻓﻈﺔ ﺿﻤﻦ اﻟﺠﻨﺲ؛ وﻣﻦ أﺟﻞ اﻟﺘﻤﻴﻴﺰ ﺑﻴﻦ اﻷﻧﻮاع ﺗﻢ‬
‫اﺳﺘﺨﺪام اﻟﻤﻨﻄﻘﺔ اﻟﺘﻲ ﺗﻠﻲ اﻟﻤﻮرﺛﺔ ‪ 16SRNA‬ﻟﻜﻮﻧﻬﺎ ﺗﺨﺘﻠﻒ ﻣﻦ ﻧﻮع إﻟﻰ ﺁﺧﺮ )اﻟﺠﺪول ‪،(1‬‬
‫وﺧﺎﺻ ًﺔ ﻟﺘﺤﺪﻳﺪ اﻟﻨﻮع ‪ L. delbrueckii subsp. bulgaricus‬واﻟﺘﻲ ﺣﺼﺮت ﺷﺪﻓﺔ ﺑﻄﻮل ‪565‬‬

‫أﺳﺎس ﺁزوﺗﻲ‪.‬‬
‫وﻳﻮﺿ ﺢ اﻟ ﺸﻜﻞ ‪ 3‬أﻧ ﻪ ﻗ ﺪ ﺟ ﺮى ﻋ ﺰل ﺟ ﻨﺲ ‪) Lactobacillus‬اﻟ ُﻌ ﺼﺎﺑﺘﺎن ‪ 5‬و ‪6‬‬

‫وﻃﻮﻟﻬﻤ ﺎ ‪ 425‬أﺳ ﺎس ﺁزوﺗ ﻲ و‪ 565‬أﺳ ﺎس ﺁزوﺗ ﻲ ﻋﻠ ﻰ اﻟﺘ ﻮاﻟﻲ( وﻧ ﻮع ‪Lb. bulgaricus‬‬

‫)اﻟﻌ ﺼﺎﺑﺔ ‪ 4‬ﺑﺤﺠ ﻢ ‪ 340‬أﺳ ﺎس ﺁزوﺗ ﻲ( ﻣ ﻦ ﺑﻌ ﺾ ﻣﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟ ﺴﻮرﻳﺔ‪ .‬وﺗﺘ ﺸﺎﺑﻪ ه ﺬﻩ‬
‫اﻟﻌﺼﺎﺋﺐ ﺑﺎﻟﺤﺠﻢ اﻟﺠﺰﻳﺌﻲ ﻣﻊ اﻟﻌ ﺼﺎﺋﺐ ‪ 1‬و‪ 2‬و‪ 3‬ﻟﻠﺠ ﻨﺲ واﻟﻨ ﻮع ‪Lactobacillus bulgaricus‬‬
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‫ﻣﻦ ﺷﺮآﺔ هﺎﻧﺴﻦ – اﻟﺪاﻧﻤﺎرك اﻟ ُﻤﺴْ َﺘﺨْ َﺪﻣﺔ آﺸﺎهﺪ إﻳﺠﺎﺑﻲ‪ .‬آﻤﺎ ﻳﻼﺣﻆ ﻣﻦ اﻟﺸﻜﻞ ﻧﻔﺴﻪ أﻧ ﻪ ﻗ ﺪ ﺗ ﻢ‬
‫ﻋﺰل اﻟﺠﻨﺲ ‪) Lactobacillus‬اﻟﻌﺼﺎﺑﺘﺎن ‪ 7‬و ‪ 8‬وهﻤﺎ ﺑﺤﺠﻢ ‪ 425 bp‬و‪ 565 bp‬ﻋﻠ ﻰ اﻟﺘ ﻮاﻟﻲ(‬
‫ﻣﻦ ﻣﻨﺘﺠﺎت اﻷﻟﺒﺎن اﻟﺴﻮرﻳﺔ وﻟﻜﻨﻬﺎ ﻟﻢ ﺗﻜﻦ ﺗﺎﺑﻌﺔ ﻟﻠﻨ ﻮع ‪ Lb. bulgaricus‬وذﻟ ﻚ ﺑﻐﻴ ﺎب اﻟﻌ ﺼﺎﺑﺔ‬
‫ﻣﻦ اﻟﻤﺴﺎر ‪.9‬‬

‫‪ PCR‬ﺑﺎﺳ ﺘﺨﺪام‬ ‫اﻟ ﺸﻜﻞ )‪ .(3‬ﻧﺘ ﺎﺋﺞ اﻟ ﺮﺣﻼن اﻟﻜﻬﺮﺑ ﺎﺋﻲ ﻟﻨ ﻮاﺗﺞ ﺗﻘﻨﻴ ﺔ اﻟ ـ‬
‫ﻣﺮﺋ ﺴﺎت ﻣﻮرﺛ ﺔ ‪ 16sRNA‬واﻟ ﺸﺪﻓﺔ اﻟﺘ ﻲ ﺗﻠﻴﻬ ﺎ‪ 1 :‬و ‪ 2‬و ‪Lactobacillus delbrueckii ssp. :3‬‬
‫‪ bulgaricus‬ﻣﻦ ﺷﺮآﺔ هﺎﻧﺴﻦ – اﻟ ﺪاﻧﻤﺎرك‪ :‬ﺷ ﺎهﺪ إﻳﺠ ﺎﺑﻲ‪ 4 ،‬و ‪ 5‬و ‪Lactobacillus delbrueckii :6‬‬
‫‪ ssp. bulgaricus‬اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ ﻣﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟ ﺴﻮرﻳﺔ‪ 7 ،‬و ‪ 8‬و ‪Lactobacillus delbrueckii : 9‬‬
‫‪ ssp. lactis‬اﻟﻤﻌﺰوﻟﺔ ﻣﻦ ﻣﻨﺘﺠﺎت اﻷﻟﺒﺎن اﻟﺴﻮرﻳﺔ‪ :MW .‬اﻟﻮاﺳﻢ اﻟﺠﺰﻳﺌﻲ‪.‬‬

‫ﺟﺮى اﻟﻠﺠﻮء إﻟﻰ ﺗﻘﻨﻴﺔ اﻟـ ‪ PCR‬ﻹﺟﺮاء ﺗﻨﻤﻴﻂ ﺟﺰﻳﺌﻲ ﻟﻠﻤﻜﻮرات ﻣﻮﺟﺒﺔ اﻟﻐ ﺮام‪ ،‬ﺳ ﺎﻟﺒﺔ‬
‫اﻟﻜﺎﺗﺎﻻز‪ ،‬اﻟﻨﺎﻣﻴﺔ ﻋﻠﻰ ﺑﻴﺌﺔ اﻟـ ‪ M17‬ﺑﻌ ﺪ اﻟﺤ ﻀﻦ ﻓ ﻲ اﻟﺪرﺟ ﺔ ‪ْ 25‬م ﻟﻤ ﺪة ‪ 48‬ﺳ ﺎﻋﺔ‪ ،‬ﻟﻠﻤﻨﻄﻘ ﺔ ﻣ ﺎ‬
‫ﺑ ﻴﻦ اﻟﻤ ﻮرﺛﺘﻴﻦ ‪ 16SRNA‬و‪ 23SRNA‬ﺑﻬ ﺪف ﺗﻤﻴﻴ ﺰ اﻟ ﺴﻼﻻت اﻟﺘﺎﺑﻌ ﺔ ﻟﻠﻨ ﻮع ‪Lactococcus‬‬

‫‪ - lactis‬ﻷﻧﻬﺎ ﻣﻨﺎﻃﻖ ﻣﺤﺎﻓﻈﺔ ﻓﻲ هﺬا اﻟﻨﻮع‪ -‬ه ﺬﻩ اﻟ ُﻤ َﺮ ﱢﺋ ﺴﺎت ﺗﺤ ﺼﺮ ﺷ ﺪﻓﺔ ﺑﻄ ﻮل ‪ 680‬أﺳ ﺎس‬
‫ﺁزوﺗﻲ‪.‬‬
‫ﻳﻈﻬﺮ اﻟﺸﻜﻞ ‪ 4‬ﻋﺼﺎﺋﺐ ذات ﺣﺠﻢ ﺟﺰﻳﺌﻲ ‪ 680‬أﺳﺎس ﺁزوﺗﻲ ﺗﻘﺮﻳﺒﺎً‪ ،‬ﻟﻨﻮاﺗﺞ ﺗﻔﺎﻋ ﻞ اﻟ ـ‬
‫‪ PCR‬ﻟﻠ ﺴﻼﻻت اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﻤﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟ ﺴﻮرﻳﺔ وه ﺬا اﻟﺤﺠ ﻢ ﻳﺘﻮاﻓ ﻖ ﻣ ﻊ اﻟ ﺸﺪﻓﺔ‬
‫اﻟﻤﺤ ﺼﻮرة ﺑ ﻴﻦ ‪ 16SRNA‬و ‪ .23SRNA‬ﻣﻤ ﺎ ﻳ ﺸﻴﺮ إﻟ ﻰ أن ه ﺬﻩ اﻟ ﺴﻼﻻت ه ﻲ ﻣ ﻦ ﻧ ﻮع‬
‫‪ ،Lactococcus lactis‬وهﺬا ﻳﺘﻮاﻓﻖ ﻣﻊ ﻣﺎ ذآﺮ ﻣﻦ ﻧﺘﺎﺋﺞ اﻻﺧﺘﺒﺎرات اﻟﻜﻴﻤﻴﺎﺋﻴﺔ اﻟﺤﻴﻮﻳﺔ‪ .‬ﻟ ﻢ ﻳﻜ ﻦ‬
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 ‫ﻟﻬﺬﻩ اﻟﺘﺠﺮﺑﺔ ﺷﺎهﺪ إﻳﺠﺎﺑﻲ ﻟﺼﻌﻮﺑﺔ اﻟﺤﺼﻮل ﻋﻠﻰ ﺳﻼﻻت ﻧﻘﻴﺔ ﻣﻔﺮدة ﻣﻦ أﺣﺪ اﻟﻤﺨﺎﺑﺮ اﻟﻌﺎﻟﻤﻴ ﺔ‬
‫اﻟ ُﻤ َﻨﻤﱢﻄﺔ ﻟﻬﺬا اﻟﻨﻮع‪ .‬وآﺎﻧ ﺖ ﻧﺘﻴﺠ ﺔ اﻟ ﺮﺣﻼن اﻟﻜﻬﺮﺑ ﺎﺋﻲ ﺳ ﻠﺒﻴ ًﺔ ﻋﻨ ﺪ اﺳ ﺘﺨﺪام ه ﺬﻩ اﻟ ُﻤ َﺮ ﱢﺋ ﺴﺎت ﻣ ﻊ‬
‫اﻟﻨﻮع ‪) S. thermophilus‬اﻟﻤﺴﺎر ‪.(1‬‬

‫اﻟ ﺸﻜﻞ )‪ .(4‬ﻧﺘ ﺎﺋﺞ اﻟ ﺮﺣﻼن اﻟﻜﻬﺮﺑ ﺎﺋﻲ ﻟﺘﻘﻨﻴ ﺔ اﻟ ـ ‪ PCR‬ﻟ ﺴﺒﻊ ﺳ ﻼﻻت ﻣ ﻦ ﻧ ﻮع ‪Lactococcus‬‬
‫‪ lactis‬ﻣﻌﺰوﻟﺔ ﻣﻦ أﻟﺒﺎن ﺑﻌﺾ اﻟﻤﻨﺎﻃﻖ اﻟﺴﻮرﻳﺔ‪ :MW .‬اﻟﻮاﺳﻢ اﻟﺠﺰﻳﺌﻲ‪.‬‬

‫‪ 2.3‬ﻧﺘﺎﺋﺞ ﻣﻄﻴﺎﻓﻴﺔ ﺗﺤﻮﻳﻞ ﻓﻮرﻳﻴﻪ ﻟﻸﺷﻌﺔ ﺗﺤﺖ اﻟﺤﻤﺮاء )‪:(FT-IR‬‬

‫ﺗﻢ ﺗﻨﻤﻴﻂ ﻋﺰﻻت ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ واﻟﻠﺒﻦ اﻟﺮاﺋﺐ ﺑﺎﺳﺘﺨﺪام ه ﺬﻩ اﻟﺘﻘﻨﻴ ﺔ‬
‫اﻟﺤﺪﻳﺜ ﺔ‪ ،‬ﺣﻴ ﺚ ﺗ ﻢ ﺑﺪاﻳ ًﺔ اﻟﺘﻌ ﺮف ﻋﻠ ﻰ اﻟﻄﻴ ﻒ اﻟﻨ ﻮﻋﻲ ﻟﻜ ﻞ ﺟ ﻨﺲ ﻣ ﻦ ه ﺬﻩ اﻟﺒﻜﺘﻴﺮﻳ ﺎ‪ُ .‬ﻳﻼﺣ ﻆ ﺿ ﻤﻦ‬
‫اﻟﻄﻴﻒ ﺛﻼث ﻣﻨﺎﻃﻖ ﺗﻤﻴﺰ ﺟﻨﺴًﺎ ﺑﻜﺘﻴﺮﻳًﺎ ﻋﻦ ﺁﺧﺮ‪:‬‬
‫‪ :cm-1 1500-1200 (1‬ﻣﻨﻄﻘﺔ ﻣﺘﻨﻮﻋﺔ‪ :‬ﺑﺮوﺗﻴﻨﺎت – ﺣﻤﻮض دهﻨﻴﺔ – ﻣﺮآﺒﺎت ﻓﻮﺳﻔﺎﺗﻴﺔ‪.‬‬
‫‪ :cm-1 1800-1500 (2‬ﻣﻨﻄﻘﺔ اﻷﻣﻴﺪ ‪ amide‬وﺧﺎﺻﺔ رواﺑﻂ أﻣﻴﺪ ‪ Ι‬وأﻣﻴﺪ ‪ II‬ﻟﻠﺒﺮوﺗﻴﻨﺎت واﻟﺒﺒﺘﻴﺪات‪.‬‬
‫‪ : cm-1 3000-2700 (3‬ﻣﻨﻄﻘﺔ ﻣﺘﻌﺪد اﻟﺴﻜﺎرﻳﺪ اﻟﻠﻴﺒﻴﺪي‪.‬‬
‫ﻳﻮﺿﺢ اﻟﺸﻜﻞ ‪ 5‬اﻟﻄﻴﻒ اﻟﻨﻮﻋﻲ ﻟﺠﻨﺲ ‪.Weissella‬‬

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 ‫اﻟﺸﻜﻞ ‪ .5‬ﻃﻴﻒ اﻟـ ‪ cm-1 3500-500 FT-IR‬ﻟﺒﻜﺘﻴﺮﻳﺎ ‪.Weissella‬‬
‫آﻤ ﺎ ﻳﻮﺿ ﺢ اﻟ ﺸﻜﻼن ‪ 6‬و‪ 7‬اﻟﻄﻴ ﻒ اﻟﻤﻤﻴ ﺰ ﻟﺒﻜﺘﻴﺮﻳ ﺎ اﻟﻤﻜ ﻮرات اﻟﻠﺒﻨﻴ ﺔ ‪Lactococcus‬‬

‫و‪ Streptococcus‬ﻋﻠﻰ اﻟﺘﻮاﻟﻲ‪.‬‬

‫وﺗﻮﺿﺢ اﻷﺷﻜﺎل ‪ 8‬و‪ 9‬و‪ 10‬ﻃﻴﻒ ﺑﻜﺘﻴﺮﻳﺎ اﻟـ ‪ Enterococcus‬وﻃﻴﻔﻲ ﺑﻜﺘﻴﺮﻳﺎ اﻟﻌﺼﻴﺎت اﻟﻠﺒﻨﻴﺔ‬
‫‪ Leuconostoc‬و‪ Lactobacillus‬ﻋﻠﻰ اﻟﺘﻮاﻟﻲ‪.‬‬

‫اﻟﺸﻜﻞ ‪ .6‬ﻃﻴﻒ اﻟـ ‪ cm-1 3500-500 FT-IR‬ﻟﺒﻜﺘﻴﺮﻳﺎ اﻟـ ‪.Lactococcus‬‬

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‫اﻟﺸﻜﻞ ‪ .7‬ﻃﻴﻒ اﻟـ ‪ cm-1 3500-500 FT-IR‬ﻟﺒﻜﺘﻴﺮﻳﺎ اﻟـ ‪.Streptococcus‬‬

‫اﻟﺸﻜﻞ ‪ .8‬ﻃﻴﻒ اﻟـ ‪ cm-1 3500-500 FT-IR‬ﻟﺒﻜﺘﻴﺮﻳﺎ اﻟـ ‪.Enterococcus‬‬

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 ‫اﻟﺸﻜﻞ ‪ .9‬ﻃﻴﻒ اﻟـ ‪ cm-1 3500-500 FT-IR‬ﻟﺒﻜﺘﻴﺮﻳﺎ اﻟـ ‪.Leuconostoc‬‬

‫اﻟﺸﻜﻞ ‪ .10‬ﻃﻴﻒ اﻟـ ‪ cm-1 3500-500 FT-IR‬ﻟﺒﻜﺘﻴﺮﻳﺎ اﻟـ ‪.Lactobacillus‬‬

‫وﻣ ﻦ اﻟﺠ ﺪﻳﺮ ﺑﺎﻟ ﺬآﺮ‪ ،‬أن ه ﺬﻩ اﻟﻨﺘ ﺎﺋﺞ آﺎﻧ ﺖ ﻣﺘﻄﺎﺑﻘ ًﺔ ﻣ ﻊ ﻧﺘ ﺎﺋﺞ ﺗﻨﻤ ﻴﻂ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ‬
‫ﺑﺎﺳﺘﺨﺪام ﺗﻘﻨﻴﺔ اﻟـ ‪ ،PCR‬آﻤﺎ أﻧﻬﺎ ﻣﺘﻄﺎﺑﻘﺔ ﻣﻊ ﻧﺘﺎﺋﺞ اﻻﺧﺘﺒﺎرات اﻟﻜﻴﻤﻴﺎﺋﻴﺔ اﻟﺤﻴﻮﻳﺔ‪.‬‬
‫ﻳﻮﺿﺢ اﻟﺸﻜﻞ ‪ 11‬ﺗﻄﺎﺑﻖ أﻃﻴﺎف ﻋﺰﻻت اﻟـ ‪ Streptococcus‬اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ واﻟﺠﺒﻨﺔ‬
‫اﻟﺒﻠﺪﻳﺔ‪ .‬ﺑﻴﻨﻤﺎ ﻳُﻈﻬﺮ اﻟﺸﻜﻞ ‪ 12‬اﺧﺘﻼف ﻃﻴﻔﻲْ اﻟـ ‪ Streptococcus‬اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋ ﺐ اﻟ ﺴﻮري‬
‫وﺗﻠﻚ اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ ﻓﻲ أﻟﻤﺎﻧﻴﺎ )ﺳﻼﻟﺔ ﻋﻴﺎرﻳﺔ(‪.‬‬

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 ‫اﻟﺸﻜﻞ ‪ .11‬ﺗﻄﺎﺑﻖ أﻃﻴﺎف ﻋﺰﻻت اﻟـ ‪ Streptococcus‬اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ واﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ‪.‬‬

‫اﻟﺸﻜﻞ ‪ .12‬اﺧﺘﻼف ﻃﻴﻔﻲ اﻟـ ‪ Streptococcus‬اﻟﺴﻮرﻳﺔ )اﻷﺣﻤﺮ( واﻷﻟﻤﺎﻧﻴﺔ )اﻷزرق(‪.‬‬

‫ﻳًﻈﻬﺮ اﻟﺸﻜﻞ ‪ 13‬ﺗﻄﺎﺑﻖ ﺳﻼﻟﺔ اﻟـ ‪ Lactococcus‬اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋ ﺐ اﻟ ﺴﻮري )اﻷﺣﻤ ﺮ(‬
‫ﻣﻊ اﻟﺴﻼﻟﺔ اﻟﻌﻴﺎرﻳﺔ اﻷﻟﻤﺎﻧﻴ ﺔ )اﻷزرق(‪ .‬أﻣ ﺎ اﻟ ﺸﻜﻞ ‪ 14‬ﻓﻴﻈﻬ ﺮ اﺧ ﺘﻼف ﻃﻴ ﻒ اﻟ ﺴﻼﻟﺔ اﻟ ﺴﻮرﻳﺔ ﻋ ﻦ‬
‫ﻃﻴﻒ اﻟﺴﻼﻟﺔ ذاﺗﻬﺎ ﻣﻦ ﻣﺎرآﺔ هﺎﻧﺴﻦ اﻟﺪاﻧﻤﺎرآﻴﺔ )ﺷﺎهﺪ ﻋﻴﺎري(‪.‬‬
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 ‫اﻟﺸﻜﻞ ‪ .13‬ﺗﻄﺎﺑﻖ ﻃﻴﻔﻲ اﻟـ ‪Lactococcus‬اﻟﺴﻮرﻳﺔ )اﻷﺣﻤﺮ( واﻷﻟﻤﺎﻧﻴﺔ )اﻷزرق(‪.‬‬

‫اﻟﺸﻜﻞ ‪ .14‬اﺧﺘﻼف ﻃﻴﻔﻲ اﻟـ ‪Lactococcus‬اﻟﺴﻮرﻳﺔ )اﻷﺣﻤﺮ( وهﺎﻧﺴﻦ اﻟﺪاﻧﻤﺎرآﻴﺔ )اﻷزرق(‪.‬‬

‫آﻤﺎ ﻟﻮﺣﻆ ﺗﻄﺎﺑﻖ ﺟﻨﺲ اﻟـ ‪ Enterococcus‬اﻟﺴﻮرﻳﺔ واﻷﻟﻤﺎﻧﻴﺔ )اﻟﺸﻜﻞ ‪.(15‬‬

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 ‫اﻟﺸﻜﻞ ‪ .15‬ﺗﻄﺎﺑﻖ ﻃﻴﻔﻲ اﻟـ ‪ Enterococcus‬اﻟﺴﻮرﻳﺔ )اﻷﺣﻤﺮ( واﻷﻟﻤﺎﻧﻴﺔ )اﻷزرق(‪.‬‬

‫ﻳﻮﺿﺢ اﻟﺸﻜﻞ ‪ 16‬اﻻﺧﺘﻼف ﺑﻴﻦ ﻃﻴ ﻒ ﺟ ﻨﺲ اﻟ ـ ‪ Lactobacillus‬اﻟ ﺴﻮرﻳﺔ وذﻟ ﻚ اﻟﻌﺎﺋ ﺪ ﻟﻠﺠ ﻨﺲ‬
‫ذاﺗﻪ ﻣﻦ اﻟﻤﺎرآﺔ اﻟﺪاﻧﻤﺎرآﻴﺔ هﺎﻧﺴﻦ‪.‬‬

‫اﻟﺸﻜﻞ ‪ .16‬اﺧﺘﻼف ﻃﻴﻔﻲ اﻟـ ‪ Lactobacillus‬اﻟﺴﻮرﻳﺔ )اﻷﺣﻤﺮ( وهﺎﻧﺴﻦ اﻟﺪاﻧﻤﺎرآﻴﺔ )اﻷزرق(‪.‬‬

‫ﻳُﻼﺣﻆ ﻣﻦ اﻟﺸﻜﻞ ‪ 17‬اﺧﺘﻼف أﻃﻴﺎف اﻟـ ‪ Leuconostoc‬اﻟﺴﻮرﻳﺔ واﻷﻟﻤﺎﻧﻴﺔ واﻟﺪاﻧﻤﺎرآﻴﺔ‪.‬‬

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‫اﻟ ﺸﻜﻞ ‪ .17‬اﺧ ﺘﻼف أﻃﻴ ﺎف اﻟ ـ ‪ Leuconostoc‬اﻟ ﺴﻮرﻳﺔ )اﻷﺣﻤ ﺮ(وهﺎﻧ ﺴﻦ اﻟﺪاﻧﻤﺎرآﻴ ﺔ )اﻟﺰه ﺮ( واﻷﻟﻤﺎﻧﻴ ﺔ‬
‫)اﻷزرق(‪.‬‬

‫ُﻳﻼﺣﻆ ﻣﻤﺎ ﺳﺒﻖ‪ ،‬أﻧﻪ ﻋﻠﻰ اﻟﺮﻏﻢ ﻣﻦ ﺗﻄﺎﺑﻖ اﻷﺟﻨﺎس واﻷﻧ ﻮاع )اﻟ ﺴﻮرﻳﺔ واﻟﻌﻴﺎرﻳ ﺔ( ﺑﺎﺳ ﺘﺨﺪام‬
‫اﻟـ ‪ PCR‬واﻻﺧﺘﺒﺎرات اﻟﻜﻴﻤﻴﺎﺋﻴﺔ اﻟﺤﻴﻮﻳﺔ‪ ،‬إﻻ أن اﻟﺒﺼﻤﺔ اﻟﻮراﺛﻴ ﺔ اﻟﺘ ﻲ ﻳﻈﻬﺮه ﺎ ﺟﻬ ﺎز اﻟ ـ ‪ FT-IR‬ﻗ ﺪ‬
‫ﺗﺘﻄﺎﺑﻖ )آﻤﺎ ﻓﻲ اﻟﺸﻜﻠﻴْﻦ ‪ (15 ،13‬وﻗﺪ ﺗﺨﺘﻠ ﻒ )آﻤ ﺎ ﻓ ﻲ اﻷﺷ ﻜﺎل ‪ ،(17 ،16 ،14 ،12‬وﻧﻌﺘﻘ ﺪ أن ﻟﻜ ﻞ‬
‫ﻣﻦ اﻻﺧﺘﻼف واﻟﺘﻄﺎﺑﻖ أهﻤﻴﺔ آﺒﻴﺮة ﻣﻦ اﻟﻨﺎﺣﻴﺔ اﻟﺘﻄﺒﻴﻘﻴﺔ ﻓﻲ ﺻﻨﺎﻋﺔ اﻷﻟﺒﺎن‪.‬‬
‫هﻨﺎك ﻣﻌﺎﻳﺮة أﺗﻮﻣﺎﺗﻴﻜﻴﺔ ﺿ ﻤﻦ اﻟﺒﺮﻧ ﺎﻣﺞ اﻟﺤﺎﺳ ﻮﺑﻲ ‪ُ OPUS‬ﺗ ﺴﻤﻰ ‪ Quality Test‬ﺗ ﺴﻤﺢ ﺑﻤﻘﺎرﻧ ﺔ‬
‫اﻟﺨ ﺼﺎﺋﺺ اﻟﻤﺨﺘﻠﻔ ﺔ ﻟﻸﻃﻴ ﺎف اﻟﻤﺪروﺳ ﺔ‪ :‬اﻻﻣﺘ ﺼﺎﺻﻴﺔ‪ ،‬اﻹﺷ ﺎرة‪/‬اﻟ ﻀﺠﻴﺞ‪... ،‬إﻟ ﺦ؛ وﺑﺎﺳ ﺘﺨﺪام‬
‫‪ Ward's algorthim‬ﻳﻈﻬﺮ اﻟﺸﻜﻞ اﻟﻌﻨﻘﻮدي ﻟﻴﻮﺿﺢ ﺗﻘ ﺎرب أو ﺗﺒﺎﻋ ﺪ اﻟ ﺴﻼﻻت اﻟﺒﻜﺘﻴﺮﻳ ﺔ اﻟﻤﺪروﺳ ﺔ‪.‬‬
‫ﻳﻮﺿ ﺢ اﻟ ﺸﻜﻞ ‪ ،18‬ﻣﻌﻄﻴ ﺎت اﻟ ـ ‪ 32‬ﻃﻴﻔ ًﺎ ﻣﺨﺘﻠﻔ ًﺎ ﻟ ﺴﻼﻻت ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ اﻟﺘﺎﻟﻴ ﺔ‪:‬‬
‫‪.Weissella ،Leuconostoc ،Streptococcus ،Enterococcus ،Lactococcus ،Lactobacillus‬‬

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 ‫اﻟ ﺸﻜﻞ ‪ .18‬ﺗﺤﻠﻴ ﻞ اﻟ ﺸﻜﻞ اﻟﻌﻨﻘ ﻮدي ‪ cluster‬ﻟ ـ ‪ 32‬ﻃﻴ ﻒ ‪ FT-IR‬ﺗﺨ ﺺ ﺳ ﻼﻻت ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ‬
‫)اﻟ ﺴﻮرﻳﺔ واﻟﻌﻴﺎرﻳ ﺔ(اﻟﺘ ﻲ ﺗ ﻢ اﺳ ﺘﻨﺒﺎﺗﻬﺎ ﻟﻤ ﺪة ‪ 48‬ﺳ ﺎﻋﺔ‪ ،‬ﻣﺠ ﺎﻻت اﻟﻄﻴ ﻒ ‪ 3000-2800cm-1‬و‪1800-‬‬
‫‪ 1500cm-1‬و‪ 1200-1500cm-1‬و ‪ 1200-900cm-1‬و ‪ .900-700-1‬وﺿﻌﺖ أﻃﻴﺎف آﻞ اﻷﻧ ﻮاع ﻣﻌ ًﺎ وﺗ ﻢ‬
‫ﺗﻌﺮﻳﻔﻬﺎ‪.‬‬
‫‪ :2-1‬ﺳﻼﻟﺘﻲ ‪ Lactobacillus‬اﻟﻌﻴﺎرﻳﺘﻴﻦ‪.‬‬
‫‪ :5-3‬ﺳﻼﻻت ‪ Lactobacillus‬اﻟﺴﻮرﻳﺔ‪.‬‬
‫‪ :6‬ﺳﻼﻟﺔ ‪ Lactococcus‬اﻟﻌﻴﺎرﻳﺔ‪.‬‬
‫‪ :11-7‬ﺳﻼﻻت ‪ Lactococcus‬اﻟﺴﻮرﻳﺔ‪.‬‬
‫‪ :14-12‬ﺳﻼﻻت ‪ Enterococcus‬اﻟﻌﻴﺎرﻳﺔ‪.‬‬
‫‪ :17-15‬ﺳﻼﻻت ‪ Enterococcus‬اﻟﺴﻮرﻳﺔ‪.‬‬
‫‪ :20-18‬ﺳﻼﻻت ‪ Streptococcus‬اﻟﻌﻴﺎرﻳﺔ‪.‬‬
‫‪ :23-21‬ﺳﻼﻻت ‪ Streptococcus‬اﻟﺴﻮرﻳﺔ‪.‬‬
‫‪ :24‬ﺳﻼﻟﺔ ‪ Leuconostoc‬اﻟﻌﻴﺎرﻳﺔ‪.‬‬
‫‪ :27-25‬ﺳﻼﻻت ‪ Leuconostoc‬اﻟﺴﻮرﻳﺔ‪.‬‬
‫‪ :28‬ﺳﻼﻟﺔ ‪ Weissella‬اﻟﻌﻴﺎرﻳﺔ‪.‬‬
‫‪ :32-29‬ﺳﻼﻻت ‪ Weissella‬اﻟﺴﻮرﻳﺔ‪.‬‬
‫ﺗ ﻢ ﺗﺤﻠﻴ ﻞ اﻟﻨﺘ ﺎﺋﺞ‪ :‬ﻣﻌﺎﻣ ﻞ اﻟﺘﺤﻠﻴ ﻞ‪ ،‬اﻟ ﺸﻜﻞ اﻟﻌﻨﻘ ﻮدي‪ ،‬اﻻﺷ ﺘﻘﺎق‪ ،‬اﻟﻄﺮﻳﻘ ﺔ‪ ،‬ﺑﺎﺳ ﺘﺨﺪام اﻟﺒﺮﻧ ﺎﻣﺞ اﻟﺤﺎﺳ ﻮﺑﻲ ‪OPUS‬‬
‫)‪ Bruker‬أﻟﻤﺎﻧﻴﺎ(‪.‬‬
‫ﺣ ﻆ وﺟ ﻮد ‪ 23‬ﻣﺠﻤﻮﻋ ﺔ‬ ‫ﻧﻼﺣﻆ أن اﻟﺸﻜﻞ ﺗﺤﺖ اﻟﻌﻨﻘ ﻮدي ﻳﺘﻔ ﻖ ﻣ ﻊ اﺧ ﺘﻼف اﻷﺟﻨ ﺎس‪ ،‬ﺣﻴ ﺚ ﻳُﻼ َ‬
‫)ﻣ ﻦ ‪ 1‬إﻟ ﻰ ‪ 23‬ﻓ ﻲ اﻟ ﺸﻜﻞ ‪ (18‬أو ﻧﻤ ﻂ ﻃﻴﻔ ﻲ ﻟﻜ ﻞ ﻣ ﻦ اﻟ ـ ‪ Lactobacillus‬و‪Lactococcus‬‬

‫ﺴﻤَﺖ‬
‫و‪ Streptococcus‬و‪ُ ،Enterococcus‬ﺗ َﻮزﱠع آﻤ ﺎ ﻳﻠ ﻲ‪ :‬ﺧﻤ ﺲ ﺳ ﻼﻻت ه ﻲ ‪ُ ،Lactobacillus‬ﻗ ﱢ‬
‫ﺑ ﺪورهﺎ إﻟ ﻰ ﺷ ﻌﺒﺘﻴْﻦ هﻤ ﺎ‪ :‬اﻟ ﺴﻼﻟﺔ اﻟﻌﻴﺎرﻳ ﺔ واﻟﻌ ﺰﻻت اﻟ ﺴﻮرﻳﺔ‪ .‬آﻤ ﺎ ﻳﻼﺣ ﻆ وﺟ ﻮد ﺳ ﺘﺔ أﻃﻴ ﺎف ﻟﻠ ـ‬
‫‪ Lactococcus‬وﺳ ﺘﺔ أﻃﻴ ﺎف ﻟﻠ ـ ‪ Enterococcus‬وﺳ ﺘﺔ أﻃﻴ ﺎف ﻟﻠ ـ ‪ Streptococcus‬ﺗﻮﺟ ﺪ ﻓ ﻲ اﻟﻔ ﺮع‬
‫اﻟﺜ ﺎﻧﻲ )ﻣ ﻦ ‪ 6‬إﻟ ﻰ ‪ 23‬ﻓ ﻲ اﻟ ﺸﻜﻞ ‪ .(18‬ﺑﻴﻨﻤ ﺎ ﻳﺤﺘ ﻮي اﻟ ﺸﻜﻞ ﺗﺤ ﺖ اﻟﻌﻨﻘ ﻮدي اﻟﺜ ﺎﻧﻲ )وإﻧﻤ ﺎ ﻓ ﻲ ﻓ ﺮع‬

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‫ﻣﻨﻔﺼﻞ( ﻋﻠﻰ ﻋﺰﻻت اﻟـ ‪) Leuconostoc‬ﻣﻦ ‪ 24‬إﻟﻰ‪ 27‬ﻓﻲ اﻟﺸﻜﻞ ‪ (18‬وﻋﺰﻻت اﻟـ ‪) Weissella‬ﻣ ﻦ‬
‫‪ 28‬إﻟﻰ ‪ 32‬ﻓﻲ اﻟﺸﻜﻞ ‪.(18‬‬
‫ﻳﻮﺿﺢ اﻟﺸﻜﻞ ‪ 19‬ﺗﺤﻠﻴﻞ اﻟﺸﻜﻞ اﻟﻌﻨﻘﻮدي ﻷﻃﻴﺎف ﺳﻼﻻت اﻟﻌﺼﻴﺎت واﻟﻤﻜﻮرات اﻟﻠﺒﻨﻴﺔ اﻟ ﺴﻮرﻳﺔ‬
‫)اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ واﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ( وﺗﻠﻚ اﻟﻌﻴﺎرﻳﺔ )هﺎﻧﺴﻦ اﻟﺪاﻧﻤﺎرآﻴﺔ واﻷﻟﻤﺎﻧﻴﺔ(‪.‬‬
‫ﻳُﻼﺣﻆ ﻣﻦ اﻟﺸﻜﻞ اﻟﺴﺎﺑﻖ ﺗﻘﺎرب ﺳﻼﻻت اﻟﻤﻜﻮرات اﻟﻠﺒﻨﻴﺔ اﻷﻟﻤﺎﻧﻴﺔ وهﺎﻧ ﺴﻦ )‪ ،5-1‬اﻟ ﺸﻜﻞ ‪ (19‬أآﺜ ﺮ‬
‫ﻣﻦ اﻟﻌﺰﻻت اﻟﺴﻮرﻳﺔ؛ ﺣﻴﺚ ﻧﺠﺪهﺎ ﻓﻲ ﻓﺮع ﻣﻨﻔﺼﻞ ﻋ ﻦ اﻟﻌ ﺰﻻت اﻟ ﺴﻮرﻳﺔ‪ ،‬وﻧﺠ ﺪ ه ﺬﻩ اﻷﺧﻴ ﺮة ﻓ ﻲ‬
‫ﻓﺮع ﺁﺧﺮ؛ اﻟﻤﻜﻮرات اﻟﻠﺒﻨﻴﺔ اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ )‪ ،9-6‬اﻟﺸﻜﻞ ‪ ،(19‬وﺗﻠﻚ اﻟﻤﻌﺰوﻟﺔ ﻣ ﻦ اﻟﺠﺒﻨ ﺔ‬
‫اﻟﺒﻠﺪﻳﺔ )‪ ،11-10‬اﻟﺸﻜﻞ ‪.(19‬‬

‫اﻟﺸﻜﻞ ‪ .19‬ﺗﺤﻠﻴﻞ اﻟﺸﻜﻞ اﻟﻌﻨﻘﻮدي ‪ cluster‬ﻟـ ‪ 19‬ﻃﻴﻒ ‪ FT-IR‬ﺗﺨﺺ ﺳ ﻼﻻت ﺑﻜﺘﻴﺮﻳ ﺎ ‪ Lactobacillus‬و‬
‫‪ Streptococcus‬اﻟ ﺴﻮرﻳﺔ واﻟﻌﻴﺎرﻳ ﺔاﻟﺘﻲ ﺗ ﻢ اﺳ ﺘﻨﺒﺎﺗﻬﺎ ﻟﻤ ﺪة ‪ 48‬ﺳ ﺎﻋﺔ‪ ،‬ﻣﺠ ﺎﻻت اﻟﻄﻴ ﻒ ‪3000-2800cm-1‬‬
‫و‪ 1800-1500cm-1‬و ‪ 1200-1500cm-1‬و ‪ 1200-900cm-1‬و ‪.900-700-1‬‬
‫‪ :3-1‬ﺳﻼﻻت ‪ Streptococcus‬اﻟﻌﻴﺎرﻳﺔ )هﺎﻧﺴﻦ(‪.‬‬
‫‪ :5-4‬ﺳﻼﻟﺘﻲ ‪ Streptococcus‬اﻟﻌﻴﺎرﻳﺘﻴﻦ )اﻷﻟﻤﺎﻧﻴﺔ(‪.‬‬
‫‪ :9-6‬ﺳﻼﻻت ‪ Streptococcus‬اﻟﺴﻮرﻳﺔ )ﻟﺒﻦ راﺋﺐ(‪.‬‬
‫‪ :11-10‬ﺳﻼﻟﺘﻲ ‪ Streptococcus‬اﻟﺴﻮرﻳﺔ )ﺟﺒﻨﺔ ﺑﻴﻀﺎء(‪.‬‬
‫‪ :13-12‬ﺳﻼﻟﺘﻲ ‪ Lactobacillus‬اﻟﻌﻴﺎرﻳﺘﻴﻦ )هﺎﻧﺴﻦ(‪.‬‬
‫‪ :15-14‬ﺳﻼﻟﺘﻲ ‪ Lactobacillus‬اﻟﻌﻴﺎرﻳﺘﻴﻦ )اﻷﻟﻤﺎﻧﻴﺔ(‪.‬‬
‫‪ :17-16‬ﺳﻼﻟﺘﻲ ‪ Lactobacillus‬اﻟﺴﻮرﻳﺔ )ﻟﺒﻦ راﺋﺐ(‪.‬‬
‫‪ :19-18‬ﺳﻼﻟﺘﻲ ‪ Lactobacillus‬اﻟﺴﻮرﻳﺔ )ﺟﺒﻨﺔ ﺑﻴﻀﺎء(‪.‬‬

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‫ﺗ ﻢ ﺗﺤﻠﻴ ﻞ اﻟﻨﺘ ﺎﺋﺞ‪ :‬ﻣﻌﺎﻣ ﻞ اﻟﺘﺤﻠﻴ ﻞ‪ ،‬اﻟ ﺸﻜﻞ اﻟﻌﻨﻘ ﻮدي‪ ،‬اﻻﺷ ﺘﻘﺎق‪ ،‬اﻟﻄﺮﻳﻘ ﺔ‪ ،‬ﺑﺎﺳ ﺘﺨﺪام اﻟﺒﺮﻧ ﺎﻣﺞ اﻟﺤﺎﺳ ﻮﺑﻲ ‪OPUS‬‬
‫)‪ Bruker‬أﻟﻤﺎﻧﻴﺎ(‪.‬‬

‫آﻤ ﺎ ﻳُﻼﺣ ﻆ أﻳ ﻀًﺎ ﺗﻘ ﺎرب ﺳ ﻼﻻت اﻟﻌ ﺼﻴﺎت اﻟﻠﺒﻨﻴ ﺔ اﻟﻌﻴﺎرﻳ ﺔ هﺎﻧ ﺴﻦ واﻷﻟﻤﺎﻧﻴ ﺔ )‪ ،15-12‬اﻟ ﺸﻜﻞ‬
‫‪ (19‬أآﺜ ﺮ ﻣ ﻦ ﺗﻠ ﻚ اﻟ ﺴﻮرﻳﺔ )‪ ،19-16‬اﻟ ﺸﻜﻞ ‪ (19‬اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﻠ ﺒﻦ اﻟﺮاﺋ ﺐ واﻟﺠﺒﻨ ﺔ اﻟﺒﻠﺪﻳ ﺔ ﻋﻠ ﻰ‬
‫اﻟﺘﻮاﻟﻲ‪ .‬ﻳﻮﺿﺢ اﻟﺸﻜﻞ )‪ (19‬أن اﻟﻤﻜﻮرات اﻟﻠﺒﻨﻴﺔ )اﻟﻌﻴﺎرﻳﺔ واﻟﺴﻮرﻳﺔ( ﺗﺘﻮاﺟ ﺪ ﻋﻠ ﻰ ﻓ ﺮع ﻣ ﻦ اﻟ ﺸﻜﻞ‬
‫ع ﺁﺧﺮ‪.‬‬
‫اﻟﻌﻨﻘﻮدي ﺑﻴﻨﻤﺎ ﻧﺠﺪ اﻟﻌﺼﻴﺎت اﻟﻠﺒﻨﻴﺔ ﻋﻠﻰ ﻓﺮ ٍ‬
‫ﻳﻮﺿﺢ اﻟﺸﻜﻞ ‪ 20‬ﺛﻼﺛًﺎ وﻋﺸﺮﻳﻦ ﻃﻴﻔًﺎ ﻟﺴﻼﻻت ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ اﻟﻤﻌﺰوﻟﺔ ﻣ ﻦ اﻟﻠ ﺒﻦ اﻟﺮاﺋ ﺐ‬
‫واﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ ﻣﻦ ﺑﻌﺾ ﻣﻨﺎﻃﻖ اﻟﻘﻄﺮ اﻟﻌﺮﺑﻲ اﻟﺴﻮري‪.‬‬

‫اﻟﺸﻜﻞ ‪ .20‬ﺗﺤﻠﻴﻞ اﻟﺸﻜﻞ اﻟﻌﻨﻘﻮدي ‪ cluster‬ﻟـ ‪ 23‬ﻃﻴﻒ ‪ FT-IR‬ﺗﺨﺺ ﺳﻼﻻت ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ اﻟ ﺴﻮرﻳﺔ‬
‫)ﻣﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﻠ ﺒﻦ اﻟﺮاﺋ ﺐ واﻟﺠﺒﻨ ﺔ اﻟﺒﻠﺪﻳ ﺔ( واﻟﻌﻴﺎرﻳ ﺔ ﺗ ﻢ اﺳ ﺘﻨﺒﺎﺗﻬﺎ ﻟﻤ ﺪة ‪ 48‬ﺳ ﺎﻋﺔ‪ ،‬ﻣﺠ ﺎﻻت اﻟﻄﻴ ﻒ ‪3000-‬‬
‫‪ 2800cm-1‬و‪ 1800-1500cm-1‬و ‪ 1200-1500cm-1‬و ‪ 1200-900cm-1‬و ‪.900-700-1‬‬
‫‪ :2-1‬ﺳﻼﻟﺘﻲ ‪ Enterococcus‬اﻟﺴﻮرﻳﺔ )اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ(‪.‬‬
‫‪ :4-3‬ﺳﻼﻟﺘﻲ ‪ Enterococcus‬اﻟﺴﻮرﻳﺔ )اﻟﻠﺒﻦ اﻟﺮاﺋﺐ(‪.‬‬
‫‪ :6-5‬ﺳﻼﻟﺘﻲ ‪ Streptococcus‬اﻟﺴﻮرﻳﺔ )اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ(‪.‬‬
‫‪ :8-7‬ﺳﻼﻟﺘﻲ ‪ Streptococcus‬اﻟﺴﻮرﻳﺔ )اﻟﻠﺒﻦ اﻟﺮاﺋﺐ(‪.‬‬
‫‪ :9‬ﺳﻼﻟﺔ ‪ Streptococcus‬اﻟﻌﻴﺎرﻳﺔ )اﻷﻟﻤﺎﻧﻴﺔ(‪.‬‬
‫‪ :10‬ﺳﻼﻟﺔ ‪ Lactobacillus‬اﻟﻌﻴﺎرﻳﺔ )اﻷﻟﻤﺎﻧﻴﺔ(‪.‬‬
‫‪ :11‬ﺳﻼﻟﺔ ‪ Lactobacillus‬اﻟﻌﻴﺎرﻳﺔ )هﺎﻧﺴﻦ اﻟﺪاﻧﻤﺎرآﻴﺔ(‪.‬‬
‫‪ :13-12‬ﺳﻼﻻت ‪ Lactobacillus‬اﻟﺴﻮرﻳﺔ )اﻟﻠﺒﻦ اﻟﺮاﺋﺐ(‪.‬‬
‫‪ :16-14‬ﺳﻼﻻت ‪ Lactobacillus‬اﻟﺴﻮرﻳﺔ )اﻟﻠﺒﻦ اﻟﺮاﺋﺐ(‪.‬‬
‫‪ :17‬ﺳﻼﻟﺔ ‪ Leuconostoc‬اﻟﻌﻴﺎرﻳﺔ )أﻟﻤﺎﻧﻴﺎ(‪.‬‬

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 ‫‪ :19-18‬ﺳﻼﻟﺘﻲ ‪ Leuconostoc‬اﻟﺴﻮرﻳﺔ )اﻟﻠﺒﻦ اﻟﺮاﺋﺐ(‪.‬‬
‫‪ :22-21-20‬ﺳﻼﻻت ‪ Leuconostoc‬اﻟﺴﻮرﻳﺔ )اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ(‪.‬‬
‫‪ :23‬ﺳﻼﻟﺔ ‪ Leuconostoc‬اﻟﻌﻴﺎرﻳﺔ )هﺎﻧﺴﻦ اﻟﺪاﻧﻤﺎرآﻴﺔ(‪.‬‬
‫ﺗ ﻢ ﺗﺤﻠﻴ ﻞ اﻟﻨﺘ ﺎﺋﺞ‪ :‬ﻣﻌﺎﻣ ﻞ اﻟﺘﺤﻠﻴ ﻞ‪ ،‬اﻟ ﺸﻜﻞ اﻟﻌﻨﻘ ﻮدي‪ ،‬اﻻﺷ ﺘﻘﺎق‪ ،‬اﻟﻄﺮﻳﻘ ﺔ‪ ،‬ﺑﺎﺳ ﺘﺨﺪام اﻟﺒﺮﻧ ﺎﻣﺞ اﻟﺤﺎﺳ ﻮﺑﻲ ‪OPUS‬‬
‫)‪ Bruker‬أﻟﻤﺎﻧﻴﺎ(‪.‬‬

‫ﺣﻴ ﺚ ﻳُﻼﺣ ﻆ وﺟ ﻮد ﻋ ﺰﻻت اﻟ ـ ‪ Enterococcus‬اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﺠﺒﻨ ﺔ اﻟﺒﻠﺪﻳ ﺔ )‪ (2-1‬وﻣ ﻦ اﻟﻠ ﺒﻦ‬

‫اﻟﺮاﺋﺐ )‪ (4-3‬ﻓﻲ ﻓﺮﻋﻴْﻦ ﻣﻨﻔﺼﻠﻴْﻦ ﻋﻦ ﺑﻌﻀﻬﻤﺎ‪.‬‬
‫ﻳُﻼﺣﻆ ﻣﻦ اﻟﺸﻜﻞ ذاﺗﻪ أن ﺳﻼﻟﺔ اﻟـ ‪ Streptococcus‬اﻷﻟﻤﺎﻧﻴﺔ )‪ (9‬ﺑﻌﻴﺪة ﻋﻦ ﻋ ﺰﻻت ه ﺬﻩ اﻟ ﺴﻼﻟﺔ‬
‫اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ )‪ (6-5‬أآﺜﺮ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ )‪ .(8-7‬وﻳﺒﻴﻦ اﻟ ﺸﻜﻞ )‪ (20‬أﻳ ﻀًﺎ أن ﻋ ﺰﻻت‬
‫اﻟﻌ ﺼﻴﺎت اﻟﻠﺒﻨﻴ ﺔ اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﻠ ﺒﻦ اﻟﺮاﺋ ﺐ )‪ (13-12‬أﻗ ﺮب إﻟ ﻰ اﻟ ﺴﻼﻟﺘﻴﻦ اﻟﻌﻴ ﺎرﻳﺘﻴﻦ اﻷﻟﻤﺎﻧﻴ ﺔ‬
‫واﻟﺪاﻧﻤﺎرآﻴ ﺔ )‪ (11-10‬ﻣ ﻦ ﺗﻠ ﻚ اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﺠﺒﻨ ﺔ اﻟﺒﻠﺪﻳ ﺔ )‪ .(16-14‬وﻳُﻈ ِﻬ ﺮ ه ﺬا اﻟ ﺸﻜﻞ أﻳ ﻀًﺎ أن‬
‫ﺳﻼﻟﺔ اﻟـ ‪ Leuconostoc‬اﻷﻟﻤﺎﻧﻴﺔ آﺎﻧﺖ أﻗﺮب إﻟﻰ اﻟـ ‪ Leuconostoc‬اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠ ﺒﻦ اﻟﺮاﺋ ﺐ )‪-18‬‬

‫‪(19‬؛ ﺑﻴﻨﻤ ﺎ اﻟﻌ ﺰﻻت اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ اﻟﺠﺒﻨ ﺔ اﻟﺒﻠﺪﻳ ﺔ )‪ (22-20‬آﺎﻧ ﺖ أﻗ ﺮب إﻟ ﻰ ﺳ ﻼﻟﺔ ‪Leuconostoc‬‬

‫اﻟﻌﺎﺋﺪة ﻟﻤﺎرآﺔ هﺎﻧﺴﻦ اﻟﺪاﻧﻤﺎرآﻴﺔ )اﻟﺸﻜﻞ ‪.(20‬‬

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 ‫‪ .4‬اﻟﻤﻨﺎﻗﺸﺔ‬
‫ﻟﻮﺣﻈﺖ أﺛﻨﺎء اﻟﺪراﺳﺔ ﻟﻌﺰﻻت ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ اﻟﻤﻌﺰوﻟﺔ ﻣﻦ ﻣﻨﺘﺠﺎﺗﻨﺎ اﻟﻤﺤﻠﻴﺔ اﻟﻨﺴﺐ‬
‫اﻟﻤﺌﻮﻳﺔ اﻟﺘﺎﻟﻴﺔ‪ :‬ﻧﺴﺒﺔ اﻟﻤﻜﻮرات اﻟﻠﺒﻨﻴﺔ ﻓﻲ اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ ‪ %56‬ﻣﻦ ﻣﺠﻤﻮع اﻟﻌﺰﻻت ﺣﻴﺚ آﺎﻧﺖ‬
‫ﻧﺴﺒﺔ اﻟـ ‪ Enterococcus‬هﻲ اﻷﻋﻠﻰ )‪(%53‬؛ ﺗﻠﻴﻬﺎ ﻧﺴﺒﺔ اﻟـ ‪ (%26) Streptococcus‬وأﺧﻴﺮًا‬
‫ﻳﺄﺗﻲ ﺟﻨﺲ اﻟـ ‪ .(%21) Lactococcus‬آﻤﺎ ُوﺟِﺪ أن ﻧﺴﺒﺔ اﻟـ ‪ E. faecium‬ﺑﻠﻐﺖ ‪ %37‬ووﺻﻠﺖ‬
‫ﻧﺴﺒﺔ اﻟـ ‪ E. faecalis‬إﻟﻰ ‪ .%63‬ﺑﻴﻨﻤﺎ آﺎﻧﺖ ﻧﺴﺒﺔ اﻟﻌﺼﻴﺎت اﻟﻠﺒﻨﻴﺔ ﻟﻠﺠﻨﺲ ‪%17 Lactobacillus‬‬

‫ﻓﻘﻂ‪.‬‬
‫أﻣﺎ ﻧﺴﺒﺔ اﻟﻤﻜﻮرات اﻟﻠﺒﻨﻴﺔ ﻓﻲ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ ﻓﻘﺪ ﺑﻠﻐﺖ ‪ %61‬ﻣﻦ ﻣﺠﻤﻮع اﻟﻌﺰﻻت ﺣﻴﺚ آﺎن‬
‫ﺟﻨﺲ اﻟـ ‪ Enterococcus‬هﻮ اﻟﺴﺎﺋﺪ ﺑﻨﺴﺒﺔ )‪(%66‬؛ ﺗﻼﻩ اﻟﺠﻨﺲ ‪ Lactococcus‬ﺑﻨﺴﺒﺔ )‪(%27‬؛ ﺛﻢ‬
‫اﻟﺠﻨﺲ ‪ Streptococcus‬ﺑﻨﺴﺒﺔ )‪ .(%7‬ﺗﺒﻴﻦ أﻳﻀًﺎ أن ﻧﺴﺒﺔ اﻟـ ‪ E. faecium‬وﺻﻠﺖ إﻟﻰ ‪ %64‬ﻓﻲ‬
‫ﺣﻴﻦ ﺑﻠﻐﺖ ﻧﺴﺒﺔ اﻟـ ‪ .%36 E. faecalis‬وآﺎﻧﺖ ﻧﺴﺒﺔ اﻟﻌﺼﻴﺎت اﻟﻠﺒﻨﻴﺔ ﻟﻠﺠﻨﺲ ‪Lactobacillus‬‬

‫‪) %12‬ﺣﻴﺚ اﺳﺘﻄﻌﻨﺎ ﺗﻨﻤﻴﻂ ﻧﻮﻋﻴﻦ هﻤﺎ‪ Lb. delbruecki bulgaricus:‬و ‪Lb. delbruecki lactis‬‬

‫ﺑﻮاﺳﻄﺔ اﻟـ ‪ .(PCR‬آﻤﺎ ﺣﺼﻠﻨﺎ ﻋﻠﻰ ﻋﺰﻻت ﺑﻴﻀﻮﻳﺔ ﻣﻦ اﻟﺠﻨﺲ ‪ Pediococcus‬واﻟﻨﻮع ‪P.‬‬

‫‪.pentosaceus‬‬
‫ﻟﻘﺪ آﺎﻧﺖ اﻟﻌﺰﻻت اﻟﺴﻮرﻳﺔ ﻣﻦ اﻟـ ‪ Streptococcus‬ﻣﺘﺤﻤﻠ ًﺔ ﻟﻠﻤﻠﻮﺣﺔ )ﺑﺘﺮآﻴﺰ ‪ (%6.5‬وهﺬا‬
‫ﻣﺎ ﻳﺘﻔﻖ ﻣﻊ ﻧﺘﺎﺋﺞ أﺑﻮ ﻳﻮﻧﺲ وزﻣﻼﺋﻬﺎ؛ ﺑﻴﻨﻤﺎ ﻻ ﻳﺴﺘﻄﻴﻊ هﺬا اﻟﺠﻨﺲ ﻋﺎد ًة اﻟﻨﻤﻮ ﻓﻲ ﺗﺮآﻴﺰ ﻣﻦ‬
‫آﻠﻮرﻳﺪ اﻟﺼﻮدﻳﻮم أﻋﻠﻰ ﻣﻦ ‪.(Geis et al., 2003) %4‬‬
‫اﺳﺘﺨﺪﻣﻨﺎ ﺟﻬﺎز اﻟﻤﻄﻴﺎﻓﻴﺔ ﺗﺤﺖ اﻟﺤﻤﺮاء اﻟـ ‪ FT-IR‬ﻟﺘﻨﻤﻴﻂ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ‪ LAB‬وﻗﻤﻨﺎ‬
‫ﺑﻤﻘﺎرﻧﺔ ﻧﺘﺎﺋﺞ هﺬا اﻟﺘﻨﻤﻴﻂ ﻣﻊ ﺗﻠﻚ اﻟﺘﻲ ﺣﺼﻠﻨﺎ ﻋﻠﻴﻬﺎ ﺑﺘﻘﻨﻴﺔ اﻟـ ‪ .PCR‬ﻳﺘﻤﻴﺰ ﺟﻬﺎز اﻟﻤﻄﻴﺎﻓﻴﺔ هﺬا‬
‫ﺑﺈﻋﻄﺎء ﻣﻌﻠﻮﻣﺎت ﺣﻮل اﻟﻤﺤﺘﻮى اﻟﻜﻠﻲ ﻟﻠﻤﻴﻜﺮوب ﺑﺸﻜﻞ ﻃﻴﻔﻲ‪ .‬ﺗﻌﺒﺮ هﺬﻩ اﻷﻃﻴﺎف ﻋﻦ ﺑﻨﻰ‬
‫ﻣﻌﻘﺪة ﻓﻲ اﻟﺠﺪار اﻟﺨﻠﻮي ﻟﻠﺒﻜﺘﻴﺮﻳﺎ وﻣﻦ اﻟﺼﻌﺐ ﻓﺼﻠﻬﺎ ﺑﺴﻬﻮﻟﺔ‪ .‬وﻟﻜﻦ رﻏﻢ ذﻟﻚ‪ ،‬أﻇﻬﺮت‬
‫ﺗﻘﺎرﻳﺮ ﻋﻠﻤﻴﺔ ﻋﺪﻳﺪة أﻧﻪ ﻳﻤﻜﻦ ﻟﺠﻬﺎز ‪ FT-IR‬أن ﻳﻜﻮن ﻣﻔﻴﺪًا ﻟﺘﻤﻴﻴﺰ اﻟﺠﻨﺲ واﻟﻨﻮع وﺣﺘﻰ اﻟﺴﻼﻟﺔ‬
‫)‪ .(Helm et al., 1991; Maquelin et al., 2003‬ﺣﻴﺚ أن أﻃﻴﺎف اﻟـ ‪ FT-IR‬ﻟﻠﻤﻴﻜﺮوﺑﺎت ﺗﻌﺘﻤﺪ‬
‫ﻧﻮﻋﻴًﺎ ﻋﻠﻰ اﻟﻄﺎﺑﻊ اﻟﻈﺎهﺮي واﻟﻤﻮرﺛﻲ ﻟﻠﺨﻠﻴﺔ اﻟﺒﻜﺘﻴﺮﻳﺔ‪ .‬إن أﻃﻴﺎف ﺗﺤﻮﻳﻞ ﻓﻮرﻳﻴﻪ ﻟﻸﺷﻌﺔ ﺗﺤﺖ‬
‫اﻟﺤﻤﺮاء ﻟﻠﺒﻜﺘﻴﺮﻳﺎ هﻲ آﺎﻟﺒﺼﻤﺔ اﻟﻮراﺛﻴﺔ ﻟﻠﺠﻨﺲ اﻟﺒﻜﺘﻴﺮي‪ .‬ﻟﺬا ﻳﻌﺘﻤﺪ ﺗﻨﻤﻴﻂ اﻟﺒﻜﺘﻴﺮﻳﺎ ﻋﻠﻰ‬
‫ﺻ َﻮر ﻣﻌﻘﺪة ﻟﻤﺠﻤﻮع‬
‫ﻼ‪ ،‬ﻣﻊ اﻷﺧﺬ ﺑﻌﻴﻦ اﻻﻋﺘﺒﺎر أن هﺬا اﻟﻄﻴﻒ هﻮ ُ‬
‫اﻟﻤﺠﺎل اﻟﻄﻴﻔﻲ آﺎﻣ ً‬
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‫اﻟﻤﺤﺘﻮى اﻟﻜﻴﻤﻴﺎﺋﻲ ﻟﻠﺨﻠﻴﺔ )ﺑﺮوﺗﻴﻨﺎت‪ ،‬أﻏﺸﻴﺔ‪ ،‬ﺟﺪر ﺧﻠﻮﻳﺔ‪ ،‬ﺣﻤﻮض ﻧﻮوﻳﺔ ‪ ....‬اﻟﺦ(‪ .‬وﻳﻤﻜﻦ‬
‫ﻟﺒﻌﺾ اﻟﻌﺼﺎﺋﺐ أن ﺗﻤﻴﺰ ﺑﻴﻦ ﻣﺠﻤﻮﻋﺎت وﻇﻴﻔﻴﺔ أو زﻣﺮ آﻴﻤﻴﺎﺋﻴﺔ‪ ،‬آﻤﺎ أن ﺑﻌﺾ ﻣﺠﺎﻻت‬
‫اﻷﻃﻴﺎف ﻣﺤﺪدة ﺣﺴﺐ ﻓﺤﻮﺻﺎت ﺧﺎﺻﺔ ﻟﻠﺨﻠﻴﺔ )‪ .(Mariey et al., 2001‬وﻋﻨﺪﻣﺎ ﻻ ﻳﻮﺟﺪ ﻃﻴﻒ‬
‫ﻧﻮﻋﻲ ﻟﺒﻜﺘﻴﺮﻳﺎ ﻣﺎ‪ ،‬ﻓﺈﻧﻪ ﻳﺠﺐ اﻻﻋﺘﻤﺎد ﻋﻠﻰ اﻟﺸﻜﻞ اﻟﻌﻨﻘﻮدي ‪ cluster‬ﻟﺘﺤﻠﻴﻞ اﻟﻨﺘﺎﺋﺞ‪ .‬وﻗﺪ ﻟﻮﺣﻆ‬
‫ﺗﻄﺎﺑﻖ ﺑﻴﻦ ﺗﻨﻤﻴﻂ ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ ﺑﻄﺮﻳﻘﺔ اﻟـ ‪ FT-IR‬ﻣﻊ ﺗﻘﻨﻴﺔ اﻟـ ‪ ،PCR‬ﻣﻤﺎ ﻳﺸﻴﺮ إﻟﻰ‬
‫إﻣﻜﺎﻧﻴﺔ اﺳﺘﺨﺪام اﻟﻄﺮﻳﻘﺔ اﻷوﻟﻰ ﻓﻲ هﺬا اﻟﺘﻨﻤﻴﻂ‪ .‬وﻋﻨﺪ ﺗﺤﻠﻴﻞ اﻟﻨﺘﺎﺋﺞ اﻋﺘﻤﺎدًا ﻋﻠﻰ اﻟﺸﻜﻞ‬
‫اﻟﻌﻨﻘﻮدي‪ ،‬ﻟﻮﺣﻆ ﺗﻘﺎرب ﺑﻴﻦ ﻋﺰﻻت ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠﺒﻦ اﻟﻤﻌﺰوﻟﺔ ﻣﻦ اﻟﻠﺒﻦ اﻟﺮاﺋﺐ اﻟﺴﻮري‬
‫وﺑﻴﻦ اﻟﺴﻼﻻت اﻟﻌﻴﺎرﻳﺔ )اﻷﻟﻤﺎﻧﻴﺔ واﻟﺪاﻧﻤﺎرآﻴﺔ( أآﺜﺮ ﻣﻨﻪ ﻓﻲ ﺣﺎﻟﺔ اﻟﺒﻜﺘﻴﺮﻳﺎ اﻟﻤﻌﺰوﻟﺔ ﻣﻦ‬
‫اﻟﺠﺒﻨﺔ اﻟﺒﻠﺪﻳﺔ )اﻷﺷﻜﺎل ‪(20-18‬؛ وﻗﺪ ﻳﻜﻮن ﺳﺒﺐ ذﻟﻚ هﻮ اﺳﺘﺨﺪام اﻟﺒﺎدﺋﺎت اﻟﻤﺴﺘﻮردة ﻓﻲ‬
‫ﺗﺼﻨﻴﻊ اﻟﻠﺒﻦ اﻟﻤﺤﻠﻲ‪ .‬وﻟﻘﺪ اﺳﺘﺨﺪﻣﺖ ﺗﻘﻨﻴﺔ اﻟـ ‪ FT-IR‬ﻓﻲ ﺗﻨﻤﻴﻂ ﺑﻌﺾ أﺟﻨﺎس ﺑﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ‬
‫اﻟﻠﺒﻦ )‪.(Curk et al., 1994; Kirschner et al., 2001‬‬

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 ‫‪ .5‬اﻻﺳﺘﻨﺘﺎﺟﺎت واﻟﺘﻮﺻﻴﺎت‬
‫‪ ‬ﺗﺘﻮاﺟﺪ ﻓ ﻲ ﻣﻨﺘﺠ ﺎت اﻷﻟﺒ ﺎن اﻟ ﺴﻮرﻳﺔ اﻟﺘﻘﻠﻴﺪﻳ ﺔ ﺑﻌ ﺾ أﻧ ﻮاع ﺑﻜﺘﻴ ـﺮﻳﺎ ﺣﻤ ـﺾ اﻟﻠ ﺒﻦ اﻟﻤﻬﻤ ـﺔ‬
‫اﻟﺘ ﻲ ﻳﻤﻜ ﻦ اﺳ ـﺘﺨﺪاﻣﻬﺎ آﺒﺎدﺋ ـﺎت ﻓ ﻲ ﺗ ﺼﻨﻴﻊ ه ﺬﻩ اﻟﻤﻨﺘﺠ ﺎت ﻣﺜ ﻞ ‪Lc. ،S. thermophilus‬‬

‫‪.Lb. casei ،Lb. bulgaricus ،lactis‬‬

‫‪ ‬ﺗﻌﺘﺒ ﺮ ﺗﻘﻨﻴ ﺔ اﻟ ـ ‪ PCR‬دﻗﻴﻘ ًﺔ ﻓ ﻲ ﺗﺤﺪﻳ ﺪ هﻮﻳ ﺔ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ اﻟﻤﻌﺰوﻟ ﺔ ﻣ ﻦ ﻣﻨﺘﺠ ﺎت‬
‫اﻷﻟﺒﺎن اﻟﻤﺤﻠﻴﺔ‪ .‬وﻳﻤﻜﻦ اﺳﺘﺨﺪام ﺗﻘﻨﻴﺔ اﻟـ ‪ FT-IR‬ﻓﻲ ﺗﻨﻤﻴﻂ هﺬﻩ اﻟﺒﻜﺘﻴﺮﻳﺎ‪.‬‬
‫‪ ‬ﻳﺠﺐ دراﺳﺔ أﺛﺮ اﻟﺘﻀﺎد ﻟﺒﻜﺘﻴﺮﻳﺎ ﺣﻤﺾ اﻟﻠ ﺒﻦ ﻋﻠ ﻰ اﻟﺒﻜﺘﻴﺮﻳ ﺎ اﻟﻤﻤﺮﺿ ﺔ‪ ،‬وﺗﺤﺪﻳ ﺪ اﻟ ﺼﺎدات‬
‫اﻟﺤﻴﻮﻳﺔ اﻟﺘﻲ ﺗﻨﺘﺠﻬﺎ‪.‬‬
‫‪ ‬ﻳﺠﺐ ﻋﺰل وﺗﻨﻤﻴﻂ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ ﻣ ﻦ اﻟﻨ ﻮق واﻟﻤ ﺎﻋﺰ اﻟ ﺸﺎﻣﻲ‪ ،‬ودراﺳ ﺔ دوره ﺎ ﻓ ﻲ‬
‫ﺗﺨﻤﻴﺮ اﻟﺤﻠﻴﺐ‪.‬‬

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 ‫آﻠﻤﺔ ﺷﻜﺮ‬
‫ﻳ ﺴﺮﻧﺎ ﻓ ﻲ ﻧﻬﺎﻳ ﺔ ه ﺬا اﻟﻌﻤ ﻞ‪ ،‬أن ﻧﺘﻮﺟ ﻪ ﺑﺎﻟ ﺸﻜﺮ اﻟﺠﺰﻳ ﻞ إﻟ ﻰ اﻟ ﺴﻴﺪ اﻟ ﺪآﺘﻮر اﻟﻤ ﺪﻳﺮ اﻟﻌ ﺎم ﻟﻠﻬﻴﺌ ﺔ‪،‬‬
‫ﻟﺘﺸﺠﻴﻌﻪ اﻟﻤﺴﺘﻤﺮ ﻋﻠﻰ إﺟﺮاء ﻣﺨﺘﻠﻒ اﻟﺪراﺳﺎت واﻟﺒﺤﻮث‪ ،‬آﻤﺎ ﻳﺴﺮﻧﺎ أن ﻧﺘﻮﺟﻪ ﺑﺎﻟ ﺸﻜﺮ اﻟ ﻰ‪ :‬رﺋﺎﺳ ﺔ‬
‫ﻗﺴﻢ اﻟﺒﻴﻮﻟﻮﺟﻴﺎ اﻟﺠﺰﻳﺌﻴﺔ واﻟﺘﻘﺎﻧﺔ اﻟﺤﻴﻮﻳﺔ‪ ،‬وإﻟﻰ اﻟﺴﻴﺪ اﻟﺪآﺘﻮر ﻋﺒﺪ اﻟﻘ ﺎدر ﻋﺒ ﺎدي وإﻟ ﻰ آﺎﻓ ﺔ اﻟﻌ ﺎﻣﻠﻴﻦ‬
‫ﻓﻲ ﻣﺨﺒﺮ اﻟﻤﻴﻜﺮوﺑﻴﻮﻟﻮﺟﻴﺎ واﻟﻤﻨﺎﻋﻴﺎت اﻟﺬﻳﻦ ﻟﻢ ﻳﺘﻮاﻧﻮا ﻋﻦ ﺗﻘﺪﻳﻢ اﻟﻤﺴﺎﻋﺪة ﺣﻴﺚ أﻣﻜﻨﻬﻢ ذﻟﻚ‪.‬‬

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 ‫ اﻟﻤﺮاﺟﻊ‬.6

‫ دراﺳ ﺔ ﺧ ﺼﺎﺋﺺ ﺑﻜﺘﻴﺮﻳ ﺎ ﺣﻤ ﺾ اﻟﻠ ﺒﻦ‬.2007 .‫ ﺳ ﻤﻴﺮ ﺳ ﻠﻴﻖ‬.‫ د‬،‫ ﺻ ﻴﺎح أﺑ ﻮ ﻏ ﺮة‬.‫ د‬،‫ﻋﻬ ﺪ أﺑ ﻮ ﻳ ﻮﻧﺲ‬
.‫ ﺟﺎﻣﻌﺔ دﻣﺸﻖ‬،‫ آﻠﻴﺔ اﻟﺰراﻋﺔ‬،‫ أﻃﺮوﺣﺔ دآﺘﻮراة‬.‫اﻟﻤﻌﺰوﻟﺔ ﻣﻦ ﺑﻌﺾ ﻣﻨﺘﺠﺎت اﻷﻟﺒﺎن اﻟﺴﻮرﻳﺔ‬
Aslim, B., Beyatli, Y., 2004. Antibiotic resistance and plasmid DNA contents of
Streptococcus thermophilus strains isolated from Turkish yoghurts. Turk. J. Vet.
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Ayad, E. H. E., Verheul, A., Wouters, J. T. M., & Smit, G. 2001. Population
dynamics oflactococc i from industrial, artisanal and non-dairy origins in
defined strain starters for Gouda-type cheese. International Dairy Journal, 11,
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Ayad, E. H. E., Verheul, A., Bruinenberg, P., Wouters, J. T. M., & Smit, G. 2003.
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Park, Y.-H., 2005. Development and evaluation of genomeprobing microarrays
for monitoring lactic acid bacteria. Appl. Environ. Microbiol. 71, 8825–8835.
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 SYRIAN ARAB REBABLIC
ATOMIC ENERGY COMMISSION
DAMASCUS- P.O.BOX: 6091

Final Report on Scientific Research

Department of Molecular biology and Biotechnology

Typing some of lactic acid bacteria in Syria using

PCR and FT-IR techniques

Dr. A. Al-mariri
Prof. N. D. Sharabi

AECS – PR \FRSR 416 November 2008

44