بررسي آلودگي كبدهاي گوسفند و گاو كشتار شده در كشتارگاه هاي استان البرز به كلستريديوم نواي به روش هاي كشت بيوشيميايي و PCR

Evaluation of Clostridium novyi infection in liver of sheep and cattle slaughtered in the slaughterhouses of Alborz province by culture, biochemical and PCR methods

ﮐﻠﺴﺘﺮﯾﺪﯾﻮم ﻧﻮاي ﺗﯿﭗ B ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻗﺎﻧﻘﺎرﯾﺎي ﻋﻔﻮﻧﯽ ﮐﺒﺪ ﻣﯽ ﺑﺎﺷﺪ. اﺳﭙﻮر ﺑﺎﮐﺘﺮي ﻫﻤﺮاه ﻏﺬا وارد ﺑﺪن ﺷﺪه و از ﻃﺮﯾﻖ ﺳﯿﺴﺘﻢ ﻟﻨﻔﺎوي وارد ﮐﺒﺪ ﻣﯽ ﺷﻮد. ﺑﻪ دﻟﯿﻞ ﺷﺮاﯾﻂ ﻫﯿﭙﻮﮐﺴﯿﮏ ﮐﺒﺪ زﻣﯿﻨﻪ ﺑﺮاي ﺗﺒﺪﯾﻞ ﺑﻪ ﻓﺮم روﯾﺸﯽ و ﺗﮑﺜﯿﺮ ﺑﺎﮐﺘﺮي ﻓﺮاﻫﻢ ﺷﺪه ﺑﺎﮐﺘﺮي ﻣﻘﺪار زﯾﺎدي ﺗﻮﮐﺴﯿﻦ ﺗﻮﻟﯿﺪ ﻧﻤﻮده ﮐﻪ در ﻧﻬﺎﯾﺖ ﻣﻨﺠﺮ ﺑﻪ ﻣﺮگ دام ﻣﯽﺷﻮد. ﺑﻨﺎﺑﺮاﯾﻦ ﺗﺸﺨﯿﺺ ﺗﻮﮐﺴﯿﻦ و ﺟﺪاﺳﺎزي ﺑﺎﮐﺘﺮي از دﺳﺘﮕﺎه ﮔﻮارش دام ﺑﻪ ﺧﺼﻮص ﮐﺒﺪ ﺟﻬﺖ ﺗﺸﺨﯿﺺ اﯾﻦ ﺑﯿﻤﺎري اﺳﺘﻔﺎده ﻣﯽﺷﻮد. ﺑﻪ ﻣﻨﻈﻮر ﮐﻨﺘﺮل ﺑﯿﻤﺎري ﺑﺮرﺳﯽ ﻓﺮاواﻧﯽ ﻣﯿﺰان آﻟﻮدﮔﯽ ﺿﺮوري ﻣﯽ ﺑﺎﺷﺪ. ﻫﺪف از اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺑﺮرﺳﯽ ﺗﻌﯿﯿﻦ ﺷﯿﻮع ﮐﻠﺴﺘﺮﯾﺪﯾﻮم ﻧﻮاي در ﮐﺸﺘﺎرﮔﺎه ﻫﺎي اﺳﺘﺎن اﻟﺒﺮز ﻣﯽﺑﺎﺷﺪ. در اﯾﻦ ﺗﺤﻘﯿﻖ ﺗﻌﺪاد 386 ﻧﻤﻮﻧﻪ ﮐﺒﺪ از ﮐﺸﺘﺎرﮔﺎه ﻫﺎ اﺧﺬ و ﭘﺲ از آزﻣﺎﯾﺸﺎت ﺑﺎﮐﺘﺮﯾﻮﻟﻮژي )ﮐﺸﺖ در ﻣﺤﯿﻂ اﺧﺘﺼﺎﺻﯽ ﺣﺎوي ﺧﻮن اﺳﺐ و ﻣﺤﯿﻂ ﮐﺸﺖ اﺧﺘﺼﺎﺻﯽ ﻣﺎﯾﻊ ، ﺗﺴﺖ ﺣﺮﮐﺖ و رﻧﮓآﻣﯿﺰي ﮔﺮم( و ﺑﯿﻮﺷﯿﻤﯿﺎﯾﯽ )ﺗﺨﻤﯿﺮ ﻗﻨﺪﻫﺎ، ﻟﯿﺴﯿﺘﯿﻨﺎز، ﻟﯿﭙﺎز، ژﻻﺗﯿﻨﺎز، اﻧﺪل، ﻫﻀﻢ ﺷﯿﺮ و ﮐﺎﺗﺎﻻز( ﻧﻤﻮﻧﻪ ﻫﺎ ﺗﻮﺳﻂ PCR ﺗﺄﯾﯿﺪ ﻧﻬﺎﯾﯽ ﺷﺪﻧﺪ. ﺑﺮاي اﯾﻦ ﻣﻨﻈﻮر از ﭘﺮاﯾﻤﺮ ﻃﺮاﺣﯽ ﺷﺪه از ﺗﻮﮐﺴﯿﻦ آﻟﻔﺎ اﯾﻦ ﺑﺎﮐﺘﺮي اﺳﺘﻔﺎده ﮔﺮدﯾﺪ. ﻧﺘﺎﯾﺞ اﯾﻦ ﺑﺮرﺳﯽ ﻧﺸﺎن داد ﮐﻪ ﻣﯿﺰان آﻟﻮدﮔﯽ ﺑﻪ اﯾﻦ ﺑﺎﮐﺘﺮي در ﮐﺸﺘﺎرﮔﺎهﻫﺎ 37 ﻣﻮرد )9/5٪( ﺑﻮد ﮐﻪ 33 ﻣﻮرد )89/18٪( آن آﻟﻮدﮔﯽ ﻫﻤﺰﻣﺎن ﺑﻪ ﮐﻠﺴﺘﺮﯾﺪﯾﻮم ﻧﻮاي و ﻓﺎﺳﯿﻮﻻ داﺷﺘﻨﺪ و ﺗﻨﻬﺎ در 4 ﻣﻮرد ﺑﺪون آﻟﻮدﮔﯽ ﺑﻪ ﻓﺎﺳﯿﻮﻻ اﯾﻦ ﺑﺎﮐﺘﺮي ﺟﺪاﺳﺎزي و ﺗﺸﺨﯿﺺ داده ﺷﺪ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﺧﺴﺎرات اﻗﺘﺼﺎدي ﻧﺎﺷﯽ از اﯾﻦ ﺑﯿﻤﺎري ﺑﻪ ﺻﻨﻌﺖ داﻣﭙﺮوي ﻻزم اﺳﺖ ﮐﺴﯿﻨﺎﺳﯿﻮن ﺑﻪ ﻣﻮﻗﻊ ﺻﻮرت ﮔﯿﺮد.

Clostridium novyi type B is the causative agent of liver infectious diseases. Bacterial spores enter the body along with food and through the lymph system get into the liver. Due to the hypoxic condition of the liver, bacteria converts to the vegetative form and replicates and produces a large amount of toxin, which ultimately leads to death. Therefore, the detection of toxin and bacterial isolation from animal gastrointestinal contents especially the liver are used to diagnose of disease. In order to control the disease, the frequency of Clostridium novyi isolates from infected animals in Alborz province is essential to be known. The aim of this study was to determine the prevalence of Clostridium novyi in slaughterhouses in Alborz province. In this study, 386 liver samples were collected from the slaughterhouses, then the bacteriological (culture in selective horse blood medium and specific liquid medium, motility test and gram staining) and biochemical tests (fermentation of sugars, lecithinase, lipase, gelatinase, Indol, milk digestion and catalase) were done, and the samples were finally confirmed using PCR. For this purpose, the primers were designed using alpha toxin DNA sequence. Results showed that the infection rate in the slaughterhouses was 37 (9.5%) in which 33 cases (89.18%) had concurrent contamination with Clostridium novyi and only 4 cases without Fasciola infection were detected. Due to the highly economic losses, vaccination is very urgent.