دراسة حيوية وتشخيص جزيئي لداء الاكياس العدرية في المضائف الوسطية اعتمادا على تحليل تتابع جينات rDNA - ITS1 وmtCOX1 في محافظة ذي قار == Biological Study and Molecular Identification on Hydatidosis in Intermediate Hosts Depending on Sequence Analysis for rDNA - ITS1 and mtCOX1 Genes in Thi - Qar Province
Author name:
وسام جاسم حنش
General topic:
Biology
Specific topic:
Microbiology - Parasites
Degree:
Doctorate
University:
University Of Basrah - College Of Education For Pure Sciences
Language:
Arabic
University location:
Basrah
First pages:
24T3592 - p.pdf
Abstract:
The present study was carried out in Thi - Qar province during the period from beginning of January until the end of December 2015 to investigate the prevalence of infection with hydatidosis in human and other intermediate hosts. The molecular techniques was used to detect the strain of Echinococcus granulosus. The present study diagnosed 58 infection with hydatidosis in different organs such as liver, lungs, kidney and spleen of surgically treated patients in AL - Hussein Imam Teaching Hospital inNassiriyah city, centre of Thi - Qar province. The total number of hydatid cysts was 72 cyst in different organs with total intensity of infection 1.24. The females recorded highest percentage of infection 67.24% than males 32.76 %, and the age group 21 - 30 year showed highest percentage of infection 24.14 % . The lowest percentage recorded in age group more than 60 year 5.17 %.The results of study showed that right lobe of liver was more affected 58.62 % than left lobe 13.79 %. The percentage of left lung infection was 10.34 % while right lung 8.62 %. The total average of fertile cysts in infected organs was 81.82 % and sterile cysts 18.18 %. The highest percentage of infection was recorded in house women 87.18 %. The number of recorded cases in Nassiriyah district was 21 case with percentage of infection 36.21 %.During the same period a total of 1303, 1287, 405, 90, and 294 head from sheep, cattle, buffaloes, camels and goats were examined respectively.The percentage of infection was 7.83 %, 7.69 %, 3.46 % and 2.22 % in sheep, cattle, buffaloes and camels respectively, and no infection was recorded in goats. The intensity of infection was 4.07, 3.27, 1.64 and 2 in sheep, cattle, buffaloes and camels respectively. The results of statistical analysis showed no significant differences (P????0.05) in percentage and intensity of infection.The percentage infection of liver, lungs and both together in sheep was 5.45 %, 1.23 % and 1.15 % respectively, and there was no significant differences in percentage of infection but intensity of infection differed significantly(P????0.05). The percentage of liver, lungs, both together and spleen infection in cattle were 3.11 %, 2.95 % , 1.55 % and 0.08 % respectively while intensity of infection in liver, lungs, both together and spleen was 2.5, 2.18, 7 and 1 respectively. The percentage of liver, lungs and both together infection in buffaloes were 2.22 %, 0.99 % and 0.25 % respectively while intensity of infection in liver, lungs and both together was 1.89, 1 and 2 respectively. No significant differences in percentage and intensity of infection in camels andboth percentage and intensity of infection in liver and lungs were 1.11% and 2 respectively.High significant differences was recorded between percentage infection in males and females of animals. The percentage of infection in females were 15.87 %, 13.42 %, 6.99 % and 3.45 % in sheep, cattle, buffaloes and camels respectively. Highest percentage of infection were 0.72 %, 0.18 % and 0.46 % in males of sheep, cattle and buffaloes respectively. No infection were recorded in males of camels.T he age was effect on percentage of infection and big aging groups recorded in sheep, cattle, buffaloes and camels. The present study recorded significant differences in percentage of fertile hydatid cysts in liver and lungs of animals and highest of percentage of fertility recorded in liver and lungs of sheep 84.21 % and 79.24 % respectively and lower percentage of fertility was in liver of buffaloes 11.11% and liver of cattle 14.70 %. The lower percentage of sterile hydatid cysts was in liver and lungs of sheep 15.79 % and 20.75 % respectively and highest percentage recorded in liver of buffaloes 88.88 % and spleen of cattle 100 %.S t a tistical analysis using of ANOVAs test showed no significant differences in total length, length of blade and width for large and small protoscolices hooks of hydatid cysts isolated from liver and lungs of sheep, human, cattle buffaloes, camels and hooks of sheep which is showed genetic variations (new strain).The PCR - RFLP technique was used in identification of E.granulosus strains infecting human and intermediate hosts in Thi - Qar province. The molecular study is carried out on 75 hydatid cysts collected from human, sheep, cattle, buffaloes and camels. The DNA is extracted from protoscolices of hydatid cysts isolated from infected organs. PCR - RFLP technique showed that sheep strain G1 of E.granulosus is predominant in Thi - Qar province depending on amplification of rDNA - ITS1 gene. The PCR products in all DNA samples showed 1000 bp approximately. Alu1 endonuclease with rDNA - ITS1 demonstrated two bands 800 bp and 200 bp approximately, while Rsa1 endonuclease showed two bands 655 bp and 345 bp approximately which is similar to sheep strain genotype 1.Sequence analysis for rDNA - ITS1 gene showed the presence of sheepstrain genotype 1 which is similar 100 % to sheep strain in gene bank exceptone isolate (sheep liver) was differs from other strains in position ofnucleotide sequence. It was similar 99 % for sequence of sheep strain. Thedata base of this new strain was recorded in gene bank for DNA sequence National Center for Biotechnology Information (NCBI) and European Nucleotide Archive (ENA) for the first time globally under the name Echinococcus granulosus strain WH1AW2 (GenBank : LT547814). Polymerase chain reaction technique and sequence analysis of mtCOX1 gene showed the presence of sheep strain G1 of human, sheep, cattle, buffaloes and camels. The sequence analysis showed the genetic variation in mtCOX1 gene for hydatid cyst in infected sheep represented by point mutation type transition which caused change Adenine to Guanine, and sequence data base of the sample recorded for first time in NCBI and ENA (GenBank : LC203589). Buffaloes strain G3 isolated from human lung recorded for first time in Thi - Qar province depending on comparison of samples sequences with source strains recorded in gene bank (NCBI) using "BLAST". The size of bands for mtCOX1 gene was 450 bp in all samples
