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العنوان
Molecular characterization of Hydatid cysts collected from different intermediate hosts /
المؤلف
Abo El-Ala, Sohila Mohamed El-gameel.
هيئة الاعداد
باحث / شيماء عبد الراضي فهمي
مشرف / ألفت عنتر مهدي أحمد
مشرف / وحيد محمد علي موسي
مشرف / سحر زايد عبد الموجود
الموضوع
Immunization. Mutation.
تاريخ النشر
2018.
عدد الصفحات
121 p. :
اللغة
الإنجليزية
الدرجة
الدكتوراه
التخصص
البيطري
تاريخ الإجازة
1/1/2018
مكان الإجازة
جامعة القاهرة - كلية الطب البيطري - Parasitology
الفهرس
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Abstract

Cystic echinococcosis is an important cosmopolitan parasitic zoonosis which causes a public health and economic problems in Egypt. So the incidence of Hydatid cyst (HC) infection was detected during the period from May 2015 to April 2016. It was found to be 7.5% in camels, 0.3% in cattle, 0.5% in sheep and 6.5% in donkeys. The SDS-PAGE analysis of camel crude HC fluid (cHCF) antigen gave 5 bands of molecular weights 181.0, 83.0, 70.0, 63.0 and 38.0 KDa. While partially purified HC fluid (ppHCF) Ag fractionated into 8 bands at molecular weight of 181.0, 83.0, 70.0, 63.0, 58.0, 48.0, 38.0 and 18.0 KDa. In addition to that, protoscolices (PSC) Ag separated into 4 bands at 181.0, 83.0, 70.0 and 63.0 KDa. Protective immunity against secondary hydatidosis was induced using three HC antigens (camel origin); cHCF, ppHCF and PSC antigens then challenged intra peritoneally with 2000 viable protoscolices. Humeral immune response accompany this protection was evaluated using ELISA. Percent of protection against HC was 86.0 %, 93.0 % and 88.4 % in immunized rabbits with cHCF, ppHCF and PSC antigens respectively. Antibody level in immunized group with ppHCF antigen on day 28 was higher than before immunization and was higher than that in PSC and cHCF antigen groups. This result indicated that ppHCF antigen can be used as a candidate for vaccine production. Also in this study germinal layers of 8 HC samples (3 of camels, 1 of cattle, 1 of sheep and 3 of donkeys) in addition to 3 secondary HC collected from rabbits were used as a source of DNA. PCR amplification of ITS1 gene of all extracted DNA samples showed an amplified DNA band of the same molecular size at 1115 bp on agarose gel. The Partial nucleotide sequences of ITS1 gene of all isolates were aligned and compared with the reference sequences of the genotype; G1- G8 on GenBank. The results showed that camel and rabbit samples were identified as Echinococcus canadensis genotype 6 (G6), while that of cattle and sheep belonged to Echinococcus granulosus sensu stricto (G1). Donkey isolates belonged to Echinococcus equines (G4). GenBank sequences accession numbers were MG458212, MG458213 and MG388204 for camel HC, MG786719 for cattle, MG786720 for sheep, MG458210 for donkey HC and MG458211 and MG388203 for rabbit secondary HC. Alignment of ITS1 partial nucleotide sequence of camel HC and rabbit secondary HC with G6 partial nucleotide sequence on GenBank were applied. Both camel HC and rabbit secondary HC showed the same sequence identity matrix that point out to absence of mutation in rabbit secondary HC. Only a single nucleotide substitution of adenine to guanine in position 47 (A47G) in rabbit secondary HC sequence were observed. It can be concluded that there is no mutation occurred during HC transmission from camel to rabbit.