تعیین میزان دوز کشنده و دوز موثر در سم مار شاخدار ایرانی

مدنی, رسول; رضوی, سیده مریم; گلچین فر, فریبا

doi:10.22092/vj.2018.116295.1389

تعیین میزان دوز کشنده و دوز موثر در سم مار شاخدار ایرانی

نوع مقاله : مقاله کامل

نویسندگان

¹ بخش پروتئومیکس و بیوشیمی، موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات، ترویج و آموزش کشاورزی، کرج، ایران.

² گروه میکروبیولوژی، دانشکده دامپزشکی، دانشگاه آزاد اسلامی واحد علوم و تحقیقات تهران، تهران، ایران.

³ دانش‌آموخته بیوشیمی، گروه زیست‌شناسی، دانشکده علوم پایه، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران

⁴ استادیار موسسه تحقیقات واکسن و سرم سازی رازی

10.22092/vj.2018.116295.1389

چکیده

جهت تعیین میزان دوز کشنده (LD50) سم مار شاخدار ایرانی و به دنبال آن تعیین دوز موثر (ED50) پادزهر (آنتی ونوم) آن، ابتدا غلظت پروتئین‌های سم مارشاخدار به روش Lowry و با استفاده از پروتئین استاندارد BSA تعیین گردید که میزان آن برابر با
µg/ml 1500 گزارش گردید. سپس برای اندازه‌گیری میزان کشندگی سم مار، در شش سطح، از موش‌های آزمایشگاهی تیپ BALB/c استرالیایی جنس ماده با وزن gr 20-18 استفاده گردید. همچنین جهت محاسبه نتایج به دست آمده از نرم‌افزار SPSS و برنامه تجزیه آماری Probit استفاده شدکه میزان آن برابر با µg/mouse 9/21 گزارش گردید وبا استفاده از مقدار LD50 به دست آمده و به وسیله مخلوط مقدارثابت سم حل شده در سالین 85 درصد و مقادیر متغیری از آنتی ونوم مونووالان میزان دوزموثر (effective dose) به دست آمد بطوری که ED50 برابر با µg/ml 395/6401 گزارش گردید.با توجه به میزان LD50 به دست آمده، مار شاخدار ایرانی جزء مارهای سمی و خطرناک محسوب می‌شود. ‏با توجه به میزان بالای ED50 بدست آمده، بهتر است از سرم پلی والان برای خنثی‌سازی سم این مار استفاده شود.

کلیدواژه‌ها

20.1001.1.24235407.1397.31.3.10.5

موضوعات

جانوران سمی

عنوان مقاله [English]

Determination of thelethal dose (LD50) and the effective dose (ED50) of Iranian horned viper venom

نویسندگان [English]

R. Madani ¹ ²
S.M. Razavi ³
F. Golchinfar ⁴

¹ Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. | Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.

² Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. | Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.

³ Student of M.Sc. in Biochemistry. Dept. of Biology, School of Basic Sciences, Science & Research Branch, Islamic Azad University, Tehran, Iran.

⁴ M.Sc. Biochemistry, Biotechnology. Assistant Professor Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization (AREEO), Karaj, IR Iran.

چکیده [English]

To determine the lethal dose (LD50) of Persian horned viper venom and the effective dose of this venom, primarily concentration of proteins in snake venom was measured by Lowry method and using BSA as standard protein which was reported to be 1500 µg/ml. The lethal dose at six levels was calculated on laboratory BALB/c Australian mice and female sex with weight of 18-20 gr, using SPSS software and Probit statistical analysis program, this amount was reported to be 21.9 µg/mouse. While utilizing the obtained LD50 amount and by the fixed amount of compound solved in 85% Saline and varied amounts of monovalent antivenom, the neutralizing dosage or effective dose could be obtained, in such a way that ratio of venom to antivenom was considered at one level and Australian BALB/c mice were injected with 18-20 gr of the female sex at six levels, and the achieved results were reported to be ED50 equal to 6401.4 µg/ml using SPSS software and Probit statistical analysis program. Considering the obtained LD50 amount, Persian horned viper is regarded as one of the poisonous and dangerous snakes. Considering the high obtained ED50 amount, it is better to use the polyvalent serum to neutralize the poson of this snake.

کلیدواژه‌ها [English]

protein assay
Persian horned viper venom
LD50 of Persian snake venom
effective dose

مراجع

1. Art Carter, Wyeth-A yerst Research, Chazy, NY. 1978. Using the Spearman-Karber Method to estimate the ED50. Sugi 1120-1125.
2. Bolaños, R. 1972. Toxicity of Costa Rican snake venoms for the white mouse. Am. J. Trop. Med. Hyg 21: 360–363.
3. Broadley CM. 1968.The venemous snakes in Central and South Africa. In: Bücherl, W, Buckley E, Deulofeu V (eds) Venomous Animals and Their Venoms. Academic Press, New York, London 4: 433-436.
4. Chippaux JP, GoyffonM . 1998. Venoms, antivenoms and immunotherapy. Toxicon 36: 823–46 .
5. Denis V. Anderade and Augusto S.1999. Relationship of venom ontogeny and diet in Bothrops. Departamento de Zoologia 200-204.
http://w.w.w.jstor.org/stable/3893080.
6. Gabriela D. Tanaka, Maria de Fa ´tima D. Furtado, Fernanda C. V. Portaro, Osvaldo Augusto Sant’Anna1, Denise V. Tambourgi. 2010. Diversity of Micrurus Snake Species Related to Their Venom Toxic Effects and the Prospective of Antivenom Neutralization.Journal. pntd. Volume 4 Issue 3.
7. Golchinfar, F. Madani, Rasool. Emami, T. 2015. Designing a competitive ELISA for evaluation of anti-snake venom serum potency. Veterinary journal (Pajuhesh & Sazandegi). 110: 9-16
8. Hsiang A, Davis M. 2015. The origin of snakes : revealing the ecology, behavior and evolutionary history of early snakes using genomics, phenomenics and the fossil record. BMC Evolutionary Biology,15:87.
9. Ismail M, Al-Ahaidib MS, Abdoon N, Abd-Elsalam MA. 2007. Preparation of a novel antivenom against Atractaspis and Walterinnesia venoms. Toxicon 49(1): 8-18.
10. John E, Burke and Edward A, Dennis . 2009. Phospholipase A2 structure/function, mechanism and signaling. Departments of chemistry and biochemistry and department of pharmacology of California, Sandiego 92093 – 601.
11. Koh, DC; Armagun, A; Jeaseelan, K. 2006. Snake venom components and their applications biomedicine. Cell 63: 3030-41.
12. Mary Ann K, Markwell, Suzanne M. 1978. A modification of the Lowry procedure to simplify protein determination in memberan and lipoprotein samples. 87:206-210.
13. Nalbantsoy,A; Karabay-Yavasoglu, NU; Sayim, F; Deliloglu-Gurhan, I; Arikan, H; Yildiz, MZ. 2012. Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production.Vol.18: 208-216.
14. Oukkache, N.; Lalaoui, M.; Ghalim, N. 2012. General characterization of venom from the Moroccan snakes Macrovipera mauretanica and Cerastes cerastes. J. Venom. Anim. Toxins Trop. Dis 18: 411–420.
15. Potter. 2003.History of the BALB/c family .Part of the Current Topics in Microbiology and Immunology book series. CT MICROBIOLOGY. Vol. 12: 1-5. http://link.springer.com/chapter/10.1007/978-3-642-70740-7_1.
16. RanaWaka, David G, Lallo and H Janaka de silva. 2013. Neurotoxicity in snake bite. Oct:e2302. Vol.7(10).
17. Rm Kini .2005. Structure – function relation ships and mechanism anticoagulant phospholipase A2 enzymes from snake venoms.Toxicon. Elsevier. 45(8):1147-61
18. S Lwanaga. 1979.Enzymes in snake venom. Springer. The series handbook of expremental pharmacology. Volume 52: 61-158.
19. Solano, G.; Segura, A.; Herrera, M.; Gómez, A.; Villalta, M.V.; Gutiérrez, J.M.; León, G. 2010. Study of the design and analytical properties of the lethality neutralization assay used to estimate antivenom potency against Bothrops asper snake venom. Biologicals 38:577–585.
20. Tabatabaei,M. Toofani, M. 2001.Determination of biological characteristics of Persian horned viper venom. 0ffice of planning & coordination of research affairs.p:2-6.cod:19609.
21. Theakston, R. D. G., Warrell, D. A. 2000. Crisis in snake antivenom supply for Africa. Lancet356-400.
22. World Health Organization (WHO). 1981. Progress in the Characterization of Venoms and Standardization of Antivenoms; WHO: Geneva, Switzerland.p(44).