Echinococcus isolates

Identification of Echinococcus Isolates using Molecular Genetic Techniques.84... 

Siles-Lucas, M., Felleisen, R., Cuesta-Bandera, C., Gottstein, B. and Eckert, J. (1994) Comparative genetic analysis of Swiss and Spanish isolates of Echinococcus granulosus by Southern blot hybridisation and random amplified polymorphic DNA technique. Applied Parasitology 35, 107-117. 

Marin, M., Carat, B., Pettersson, LJ. and Ehrlich, R. (1993) Isolation and characterization of a middle repetitive DNA element from Echinococcus granulosus. Molecular and Biochemical Parasitology 59, 335-338. 

A virtually untapped area for studying diversity in Echinococcus is the use of microsatellite DNA. Some microsatellite markers are available for E. multilocularis, following the earlier studies of Bretagne et al. (1996) who were able to use microsatellite DNA to divide isolates of E. multilocularis into three groups European, North American (Montana) and Japanese. More recently Nakao et al. (2003) isolated two microsatellite loci that were used to demonstrate population-level polymorphisms in E. multilocularis adult worms derived from wild red foxes collected from a limited geographical area of Hokkaido, Japan. 

Azab, M.E., Bishara, S.A., Helmy, H., Oteifa, N.M., El-Hoseiny, L.M., Ramzy, R.M. and Ahmed, MA. (2004) Molecular characterization of Egyptian human and animal Echinococcus granulosus isolates by RAPD-PCR technique, journal of the Egyptian Society of Parasitology 34, 83-96. 

Bartholomei-Santos, M.L., Heinzelmann, L.S., Oliveira, R.P., Chemale, G., Gutierrez, A.M., Kamenetzky, L., Haag, K.L. and Zaha A. (2003) Isolation and characterization of microsatellites from the tapeworm Echinococcus granulosus. Parasitology 1 26, 599-605. 

Nakao, M., Sako, Y. and Ito, A. (2003) Isolation of polymorphic microsatellite loci from the tapeworm Echinococcus multilocularis. Infection Genetics and Evolution 3, 159-163. 

Fig. 4.7. Phylogenetic tree of most available ITS1 sequences from Echinococcus species. Minimum evolution tree found in MEGAv3 as described in the text. Identical Greek letters indicate sequences of cloned PCR products from the same isolate, a and p - Lavikainen et al. (2003) y- Kedra et al. (1999) 8 - van Herwerden et al. (2000) e - Bowles et al. (1995).

Walker, M., Baz, A., Dematteis, S., Stettler, M., Gottstein, B., Schaller, J. and Hemphill, A. (2004) Isolation and characterization of a secretory component of Echinococcus multilocularis metacestodes potentially involved in modulating the host-parasite interface. Infection and Immunity 72, 527-536. 

Fig. 6.3. SDS/PAGE (5-15% linear gradient gel) separated proteins of the total worm homogenate and isolated brush border fractions from protoscoleces of Echinococcus granulosus (horse strain), (a) Coomassie blue staining (b) Periodic acid-Schiff (PAS) staining. (After McManus Barrett, 1985.)...

H. citelli, H. diminuta and H. microstoma, one of which is mitochondrial DNA (mtDNA) (394). The mtDNA of H. diminuta has been isolated (118) and has been shown to be a typical circular molecule. The characteristics of H. diminuta DNA are shown in Table 6.11. In contrast, E. multilocularis and E. granulosus produced two distinct DNA bands after fractionation in caesium chloride, but there was no evidence that the DNA from either band represented mtDNA (493). There is presumably so little mtDNA in comparison to nuclear DNA in these organisms that it is completely masked in preparations of total DNA by this method. That this is the case has been shown by a recent study (976), where a different procedure, based on the selective precipitation of nucleic acids by cetyltrimethylammonium bromide (CTAB), was employed to extract mtDNA from isolated mitochondria. Some 300 g and 50 g, respectively, of Taenia spp. and Echinococcus sp. tissue yielded approximately only 1 ng mtDNA. 

An example of the former cloning strategy, outlined in Fig. 6.17, is provided by Rishi McManus (695), who have constructed a small, size-selected genomic DNA library in a plasmid vector of Escherichia coli using total DNA isolated from Echinococcus granulosus. Subsequent differential... 

McManus, D. P. Barrett, N. J. (1985). Isolation, fractionation and partial characterization of the tegumental surface from protoscoleces of the hydatid organism, Echinococcus granulosus. Parasitology, 90 111-29. 

McManus, D. P., Knight, M. Simpson, A. J. G. (1985). Isolation and characterisation of nucleic acids from the hydatid organisms Echinococcus spp. (Cestoda). Molecular and Biochemical Parasitology, 16 251-66. 

Rishi, A. K. McManus, D. P. (1987a). Genomic cloning of human Echinococcus granulosus DNA isolation of recombinant plasmids and their use as genetic markers in strain characterization. Parasitology, 94 369-83. 

Chang GY, Ko DY. Isolated Echinococcus granulosus hydatid cyst in the CNS with severe reaction to treatment Neurology 2000 54(3) 778-9. 

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