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1.
Biol. Res ; 42(2): 147-151, 2009. ilus
Article in English | LILACS | ID: lil-524884

ABSTRACT

DNA extraction methods for genotyping non-invasive samples have led to great advances in molecular research for ecological studies, and have been particularly useful for analyzing threatened species. However, scarce amounts of fragmented DNA and the presence of Taq polymerase inhibitors in non-invasive samples are potential problems for subsequent PCR amplifications. In this study we describe a novel technique for extracting DNA from alimentary tract cells found on external surfaces of feces and regurgitated seeds. The presence of contaminants and inhibitors is minimized and samples are preserved intact for use in other ecological research (e.g. trophic studies). The amplification efficiency and purity of the extracted DNA from feces were significantly higher than in commonly used extraction procedures. Moreover, DNA of two bird species was identified from seeds expelled by regurgitation. Therefore, this method may be suitable for future ecological studies of birds, and other vertebrate groups.


Subject(s)
Animals , Columbidae/classification , DNA, Mitochondrial/isolation & purification , Feces/chemistry , Seeds/chemistry , Columbidae/genetics , Filtration/instrumentation , Genotype , Polymerase Chain Reaction
2.
Veterinary Medical Journal. 2008; 56 (4): 333-342
in English | IMEMR | ID: emr-90763

ABSTRACT

Avipoxviruses from different geographic regions of the world have been characterized to study their genetic and biological properties, but so far, no such work has been performed on Egyptian isolates. Lesions suggestive of avian pox; found on Egyptian wild dove; were used for isolation of pox virus in a previous study. The resulting virus was propagated in chorio-allantoic membrane [CAM] of specific pathogen free [SPF] Embryonated chicken eggs [ECE]. PCR was carried out on the DNA of the dove poxvirus [DPV], pigeon poxvirus [PPV] and a vaccinal strain of fowl pox virus [FPV], then restriction fragment length polymorphism [RFLP] assay was carried out on the resulting amplicons of 578 bp length; using EcoRV and NlaIII restriction enzymes. The restriction profile revealed that the dove pox virus is identical to the PPV and both are different from FPV. The results of immunoblotting analysis of the 3 pox viruses against chicken anti FPV revealed that in spite of the minor antigenic differences observed between them the DPV is closely related to the PPV. In conclusion the Egyptian wild doves are found to play a serious role in the epidemiology of PPV among pigeon flocks


Subject(s)
Animals , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Immunoblotting/methods , Columbidae/genetics , Avipoxvirus
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