Bachal Bhutto

Prevalence and genotypic characterization of bovine Echinococcus granulosus isolates by using cytochrome oxidase 1 (Co1) gene in Hyderabad, Pakistan

Cystic echinococcosis is an important zoonotic disease; it has serious impacts on animals as well as human health throughout the world. Genotypic characterization of Echinocossus granulosus (E. granulosus) in buffaloes has not been addressed in Pakistan. Therefore, the present study was conducted to evaluate the incidence and genotypic characterization of bovine E. granulosus. Out of 832 buffaloes examined, 112 (13.46%) were found infected. The favorable site for hydatid cyst development was liver (8.65%) followed by lungs (4.80%). The rate of cystic echinococcosis was found higher in females 14.43% than males 9.77%. The females above seven years aged were more infected as compared to the young ones. The partial sequence of mitochondrial cytochrome oxidase 1 (CO1) gene was used for identification and molecular analysis of buffalos E. granulosus isolates. The alignment of redundant sequences were compared with already identified 10 genotypes available at National Centre for Biotechnology Information (NCBI) GenBank. The sequencing and phylogenetic analysis of all randomly selected buffalo isolates were belong to the G1- G3 complex (E. granulosus sensu stricto). All sequences were diverse from the reference sequence. No one showed complete identity to the buffalo strain (G3), representing substantial microsequence variability in G1, G2 and G3 genotypes. We evaluated the echinococcal infectivity and first time identification of genotypes in buffaloes in Sindh, Pakistan. This study will lead to determine accurate source of this zoonotic disease to humans in Pakistan.

Effect of Temperature and Storage Time on DNA Quality and Quantity from Normal and Diseased Tissues

DNA extraction and purification is an initial step for authentic results in advance molecular biology, therefore DNA degradation is unavoidable. The aim of present study is to investigate the DNA quality and quantity in terms of shorter time preservation with normal and diseased tissue, therefore tissues of normal ( n = 18) and diseased ( n = 18) liver, lung and heart was collected from goat after slaughtered. For DNA extraction Gene JET Genomic DNA Purification Kit protocol was followed, then stored at -20 o C and -04 o C temperatures for 24hrs and 48hrs period of time. The concentration and purity of the extracted DNA were measured with Spectrophotometer and purity confirmed at an absorbance ratio of 260 or 280. It was observed that at a -20 o C temperature for 24hours the concentration of DNA yield was numerically higher than at -04 o C temperature for tissue stored at 48hrs, whereas absorbance was higher, however in normal tissues in contrast with diseased the concentration and absorbance of DNA was somehow same at -20 and -04 o C but different in storage time. On the basis of these findings, it was concluded that time elapsed between sampling with the storage condition and with normal or diseased samples for DNA extraction will largely depend on the experiment. If tissue preservative conditions and sampling are appropriate, storage time will not be a factor at least for short storage periods.

Conventional and Molecular Detection of Plasmodium in Domestic Poultry Birds

The present study was conducted to detect the Plasmodium, the causative agent of malaria in domestic poultry birds. Blood smear method was used as the conventional method for the detection, whereas the polymerase chain reaction (PCR) was performed for further confirmation. A total of 50 blood samples were collected from poultry birds showing the malarial symptoms. The results of blood smear methods showed two samples (4%) were infected with genus Plasmodium, whereas the PCR analysis showed four (8%) positive samples. These results confirm that the PCR is more sensitive method for detecting the Plasmodium when compared with conventional methods, and the microscopy diagnosed 50% false negative results that were confirmed by PCR.