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1. Breed Characterization Of Red Sindhi And Tharparkar Cattle Breeds By Mitochondrial D-Loop And Gytochrome

by Nabeela Akhtar | Mr. Tanveer Hussain | Prof. Dr. Masroor Elahi Babar.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Livestock plays an important role in economy of Pakistan. Different livestock animals used for for meat, milk, draught, and sports. The genetic data of different cattle breeds like Red Sindhi and Tharparkar is not available which needs to be established for their genetic identification, conservation and to find their genetic diversity among them. Blood samples of pure bred animals were collected from their respective home tracts. The Red Sindhi cattle samples were collected from (Barani Livestock Production Research Institute, Kherimurat, Attock, Govt. Red Sindhi Cattle Breeding Farm, Tando Muhammad Khan and Department of Livestock Management, Sindh Agriculture University, Tandojam) and Tharparkar cattle samples were collected from (Department of Livestock Management, Sindh Agriculture University, Tandojam, Tharparkar cattle Farm at Nabi Sar Road and from Tando Qaiser in Sind). DNA was extracted with the standard protocol in Molecular Biology and Genomics Laboratory of Institute of Biochemistry and Biotechnology. Specific primers were designed by using special softwares Primer 3 for mitochondrial D-loop region and Cytochrom b gene from NCBI accession no. AF492350. Then after primers optimization PCR amplification was done. Then sequencing of target fragments was carried out. Sequences were alligned with the help of software blast2sequence. Single Nucleotide Polymorphisms (SNP5) were identified and comparison of 5 mitochondrial DNA haplotypes of two cattle breeds was done. Sequences were analyzed and compared with already reported sequence of Mitochondrial DNA of Bos indicuss, Bos taurus and Bubalus bubalis available at NCBI. Phylogenetic tree was constructed using MEGA 4.1 software (http://www.megasoftware.net/MEGA4.1.html) showed that Pakistani, European and Asian cattle are genetically same but different from Buffalo. This work provided the genetic data which is very helpful for determining the genetic diversity of cattle population, breed identification, animal forensic and paternity cases and making effective breeding policies and conservational activities in future. This work is very helpful about breed characterization of two cattle breeds (Red Sindhi and Tharparker) and developing understanding about genetic architecture of cattle breeds as present study conclude that six SNPs were present in both breeds, four private to Red Sindhi and 22 were private to Tharparkar. Availability: Items available for loan: UVAS Library [Call number: 1155,T] (1).

2. Study Of Autosomal Recessive Non Syndromic Mental Retardation Locus By Linkage Analysis

by Sajjad Ali Shah | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Tanveer Hussain.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 2010Dissertation note: Mental retardation (MR) is the retarded conditions of mind in which the intelligence quotient (IQ) is lower than 70, associated with a deficiency in adaptive behavior such as communication and daily living skills. Mental retardation is either the only consistent handicap (non-syndromic) or is combined with other physical and br behavioral abnormalities (syndromic). It is one of the most common disorders and it affects about 1-3% of the human population, with a proportion higher in males than females. In the present study 10 families with two or more affected individuals were selected from different areas of Malakand Division and district Mardan of Khyber Pakhtunkhwa. Family history was taken and pedigrees were made personally by visiting the families and using specially designed proformas after their consent. The blood was collected from the selected families aseptically. Then DNA was extracted by standard inorganic protocol. Short Tandem Repeat (STR) markers (D3S3630, D3S3050, D3S1620) in vicinity of MR locus (MRT2CRBN gene) were selected, optimized and amplified by Polymerase Chain Reaction. The affected families were screened for linkage to MRT2A locus using Polyacrylamide Gel Electrophoresis (PAGE). The haplotypes were then constructed to determine the linkage of families to MRT2A locus. Out often selected families two families (MR-02 and MR-07) showed linkage to autosomal recessive nonsyndromic mental retardation locus MRT2A. This is the first report of MRT2A phenotype linkage in families from Malakand Division where consanguineous marriages are very common. Further study is needed to explore the other linkages in mentally retarded families in local population. The present study will help us to determine the genetics basis of mental retardation in affected families of Pakistan. It will also help us to screen out carrier individuals in our population that would help to develop genetic counseling strategies to prevent the progression of mental retardation in the country. Availability: Items available for loan: UVAS Library [Call number: 1162,T] (1).

3. Molecular Diversity Analysis Of Sheep And Goat Breeds Of Pakistan Using Microsatellites.

by Misbah Shaheen | Prof.Dr.Masroor Elahi Babar | Mr. Tanveer Hussain | Prof. Dr. Azhar.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Pakistan is rich in Animal Genetics Resource (AnGR) and has various breeds of sheep and goat but the genetic data in these different breeds is lacking which needs to be established for their genetic identification. The advent of molecular techniques has led to an increase in the studies that focus on the genetic characterization of domestic breeds using genetic markers. Due to their reliability and availability, the microsatellites have become preferred method for the genome mapping. Microsatellites or STRs are the 1-6 nucleotide tandem repeats present in both coding and non coding regions of both prokaryotes and eukaryotes. Microsatellites are powerful tools in genome mapping, forensic DNA studies, paternity testing, population genetics and conservation! management of biological resources. The present study was conducted on the molecular diversity analysis of sheep and goat breeds of Pakistan using FAQ recommended unlabelled microsatellites. Blood samples of unrelated true representative animals of two sheep and goat breeds were selected from their breeding tracts and different Government Livestock Farms throughout the country. DNA was extracted with the standard protocol and amplification of DNA was done with a set of 16 microsatellite markers in Molecular Cytogenetics and Genomics Laboratory in the Institute of Biochemistry and Biotechnology. The products of touch-down PCR were examined on non denaturing Polyacrylamide Gel Electrophoresis (PAGE). Genotyping results were analyzed through the sofiware POPGENE version 3.3 for calculating the number of alleles, expected and observed heterozygosity, homozygosity, Polymorphic Information Content (PlC). Average observed heterozygosity, average observed homozygosity, observed and effective number of alleles for all loci and populations were 0.8394, 0.1606, 3.6875 and 2.8693 respectively. Almost all of the microsatellite markers showed significant variations in both breeds of sheep and goat. Genotyping results of microsatellite markers were clearly different for four different breeds showing a distinct genetic distance between sheep and goat breed's. This work provided the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in future according to FAO global Farm Anithal Genetic resource data. Moreover this study can become the basis for further research investigations in sheep and goat in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1200,T] (1).

4. Genetic Study Of Ushic/Dfnb18 By Linkage Analysis

by Muneer Ahmad | Mr. Tanveer Hussain | Prof. Dr. Masroor Elahi Babar.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Deafness refers to conditions in which individuals are fully or partially unable to detect or perceive at least some frequencies of sound which can typically be heard by members of their species. In human beings, the term hearing impairment is usually reserved for people who have relative insensitivity to sound in the speech frequencies. In the present study, six families were identified which were collected from FATA, Lahore and Sheikhupura and were consist of at least two deaf people. Pedigrees of the affected families were drawn using Cyrillic software. Blood samples were collected from these families. DNA was extracted through Inorganic protocol. The creening of the affected families was done for known deafness locus, USH1C/ DFNBI8. Then the PAGE (Polyacrylamide Gel Eletrophoresis) was done and haplotypes were constructed to determine whether a family was linked to deafness locus or not. Out of six families, no family was linked to USHIC/ DFNB18 locus. Availability: Items available for loan: UVAS Library [Call number: 1223,T] (1).

5. Genetic Diversity Analysis Of Sahiwal And Dhanni Cattle Breeds By Cytochrome B Gene And Microsatellite Markers

by Zahoor Ahmed | Prof.Dr.Masroor Elahi Babar | Mr. Tanveer Hussain | Prof.Dr.Muham.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 2010Dissertation note: Pakistan has various dairy breeds of cattle but the genetic data of different cattle breeds including Sahiwal and Dhanni is lacking which need to be established for their genetic identification. Blood samples of unrelated true representative of breeds (Sahiwal and Dhanni) were collected from their respective home tracts and different Government livestock farms. DNA extracted with the standard protocol (Inorganic Method) in Molecular Biology and Genomic Laboratory, Institute of Biochemistry and Biotechnology (IBBT), University of Veterinary and Animal Sciences, Lahore. Nine fluorescent dye labeled microsatellite markers having high polymorphism information content (PIC) values were used and genotyping was done. These results were analyzed statistically by softwares "POPGENE 1.31 and POWER STAT" 2.1. Allele frequency, heterozygosity, homozygosity, polymorphism information content (PIC), power of discrimination, power of exclusion, F-Statistics and Gene Flow for all Loci, population's dendogram, Nei's genetic identity and genetic distance/ diversity were calculated. The average observed heterozygosity was 0.5845 and 0.5911 in Dhanni and Sahiwal respectively. The mean observed homozygosity was 0.4155 and 0.4089 in Dhanni and Sahiwal respectively. The average PIC (Polymorphic Information Content) values of nine loci showed by Dhanni and Sahiwal cattle are 0.61 and 0.77 respectively. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between Dhanni and Sahiwal cattle breeds. For further confirmation of the breeds amplification of the mitochondrial Cyt b gene was done with especially designed primers which were designed by using Primer3 software. Sequencing of PCR fragments was done. Analysis of the sequences was performed by multiple sequence alignment with the help of Blast 2sequence and BioEdit soft wares. Identified SNPs were analyzed and haplotypes were formed. Phylogenetic tree was constructed by MEGA 4.1. The use of genetic markers provided the information on population genetic structures of the indigenous cattle breeds even if they lack detailed pedigree recording data. The study on the genetic diversity showed the differentiation of breeds and individual breeds have unique combinations of genes as a result of phylogenetic tree. This work will provide the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in Pakistan in future according to FAO global Farm Animal Genetic resource data. Availability: Items available for loan: UVAS Library [Call number: 1289,T] (1).

6. Sequence Analysis Of Shiga Toxin 1 And Shiga Toxin 2 Genes Of Escherichia Coli O157: H7 Isikates From Lahore

by Saqib Hussain | Mr. Tanveer Hussain | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2011Dissertation note: Escherichia coli is normal inhabitant of all the animals and human beings. The Sorbitol non fermenting E. coli strains were detected in milk, beef and fecal samples collected from different areas of Lahore. White colored colonies of each positive sample on Sorbitol MacConkey's Agar (SMA) contained gram negative, rods with round ends non spore former Escherichia coli. Each of the isolates was sorbitol non fermenter, lactose fermenter, indole positive, Methyl Red positive, Voges Prauskaur negative and citrate negative. Each of the isolate was further characterized using polymerase chain reaction (PCR) for the presence of shiga toxin 1 and shiga toxin 2 genes. Bacterial DNA was extracted easily by boiling method when the isolates were grown on Sorbitol MacConkey's agar at 37°C for 12-24 hours. The DNA was recovered when the culture was boiled for 5-10 minutes. The isolated DNA when amplified using Stxland Stx2 specific primers showed that 68.5 percent samples were positive for Stxl and 54.2 percent for Stx2. The stx 1 and stx2 PCR products were subjected to sequencing. The resulted sequences when aligned with the reference sequence through Basic local alignment tool it showed that the shiga toxin 1 and shiga toxin 2 gene sequences are conserved and showed high similarity in their nucleotide structure. Despite of having high similarity in their nucleotide structure some haplotypes were also obtained showing single nucleotide polymorphism. Phylogenetic analysis made among local isolates and also with reported sequences from all over the world by using bioinformatics software to see the genetic similarities and difference between them. The data produced showed some highly conserved sequences and SNPs as well that will be quite useful for further applications in diagnostics and biotechnology applications in future. Availability: Items available for loan: UVAS Library [Call number: 1375,T] (1).

7. Molecular Characterization Of Local Isolation Of Staphylococcus Aureus On The Bsis Of 16S Rrna From Poulry And Their Transmission to Humans

by Muhammad Rizwan Ashraf | Mr. Muhammad Asif | Dr. Aby Saeed | Mr. Tanveer Hussain.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: Staphylococcus aureus is a widely distributed throughout the world and makes up the normal bacterial flora of skin and mucous membranes of man and animals. It is involved in suppurative wound infections in man and animals. Poultry industry has also been effected by S. aureus and causing great economic and health problems. The focus of the microbiology is to correctly identify S. aureus for the treatment of the animals. Molecular biology and biotechnology is proving a helping hand in the accurate identification of microorganisms through sequence analysis of 16S rRNA gene. The aim of this study was the molecular characterization of S. aureus from poultry and poultry farm workers through 16S rRNA analysis. Bacteria were collected from poultry and poultry farm human workers. All the samples were cultured and tested biochemically. In addition, peR amplification of 16S rRNA was performed in order to sequence the gene and further analyses through bioinforrnatics tools were performed. The aim of the study was the molecular characterization of S. aureus in poultry and humans through 16S rRNA sequencing, finding the phylogenetic relationships among S. aureus isolates and detection of zoonoses between poultry and human. 16S rRNA gene was amplified with peR primers and the sequence was compared with NeBI database reported S. aureus sequences. Resemblance was found between human and chicken isolates. Phylogenetic analyses were performed by using MEGA5 so ftware that also showed phylogenetic relationship among them. Availability: Items available for loan: UVAS Library [Call number: 1393,T] (1).

8. Genetic Variability Of Sahiwal And Cholistani Cattle Breeds Of Pakistan Usin Mitochondrial D-Loop Sequences

by Sania Saeed | Mr. Tanveer Hussain | Dr. Abu Saeed | Dr. Muhammad Wasim.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2011Dissertation note: Pakistan is rich in cattle genetic resources. The phenotypic and genetic diversity of animal breeds in Pakistan is very vast. Efforts to manage and utilize these genetic resources efficiently are lacking due to lack of both awareness and weakness of Government institutions. The genetic data of dairy cattle breeds (Sahiwal and Cholistani) is not yet been studied for their genetic identification, conservation and to find the genetic diversity among them and it needs to be established. For this study the blood samples( 25 samples from each breed) were collected from their home tracts and livestock farms. Unrelated animals with typical phenotypic features known for Sahiwal and Cholistani cattle breeds were selected from their breeding areas and Government livestock farms. Blood samples from true representative individuals of Sahiwal breed were collected from Research Centre for the Conservation of Sahiwal Cattle (RCCSC), Jahangirabad, Khanewal, Semen Production Unit (SPU) Qadirabad and Barani Livestock Production & Research Institute (BLPRI), Kherimurat District Attock. Cholistani cattle samples were collected from Govt. Livestock Farm, Jugaitpeer, Bahawalpur. Sampling from siblings was avoided to minimize inbreed samples as it results in depleting of gene pool along with causing inbreeding depression.DNA was extracted and quantified with the standard protocol in Molecular Biology and Genomics Laboratory of Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore. Specific primers were designed by using special software i.e. Primer Fox for mitochondrial D-loop region from NCBI accession no. NC_006853.1.Primers optimization was done after primer designing and afterwards, PCR amplification was performed. Then sequencing of target fragments was carried out using Prism ABI 3130L sequencer and Analyser.Sequences were alligned with the help of software blast2sequence and SNPs were detected. It was found that ratio of transition mutation was higher than transversions i.e. 41 transition and 10 transversions. Sequences were analyzed and compared with already reported sequence of Mitochondrial DNA of Bosindicuss, Bostaurus, Bubalusbubalis, Canis lupus familiaris, Caprahircus, Equuscaballusisolate, Ovisaries and Cameliusdromedaries sequencesavailable at NCBI. Single Nucleotide Polymorphisms (SNPs) were then detected. A phylogenetic tree constructed using MEGA 5.1 software revealed that Pakistani, European and Asian cattle are genetically same but different from Buffalo.This work is very helpful about breed characterization of two cattle breeds (Sahiwal and Cholistani) and developing understanding about genetic architecture of cattle breeds as present study conclude that 52 SNPs were present in Sahiwal and Cholistani breed of Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1498,T] (1).

9. Assessment Of Genetic Diversity In Balochi And Rakhshani Sheep Breeds Of Balochistan Using Microsatellite Dna

by Abdul Wajid | Dr. Muhammad Wasim | Dr. Abu Saeed | Mr. Tanveer Hussain.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Pakistan being agriculture based country has a great potential in livestock sector, it plays an important role in the economy of the country. Pakistan is rich in Animal Genetics Resource (AnGR) and has various breeds of sheep but lacking genetic data of these breeds which need to established data for their genetic identification. Customarily, classification of breed was based on phenotypic traits. In some cases, recent genetic studies have found differences in the structure proposed. Molecular characterization is a prevailing tool to consider the genetic variation existed within and among breeds. Characterization and evaluation of genetic differences among these breeds is necessary for their effective and meaningful improvement and conservation. The advent of molecular techniques has led to an increase in the studies that focus on the genetic characterization of domestic breeds using genetic markers. Due to their reliability and availability, the microsatellites have become preferred method for the genome mapping. Microsatellites or STRs are the 2-6 nucleotide tandem repeats present in both coding and non coding regions of both prokaryotes and eukaryotes. Microsatellites are powerful tools in genome mapping, forensic DNA studies, paternity testing, population genetics and conservation/ management of biological resources. The present study was conducted on the molecular diversity analysis of two sheep breeds Balochi and Rakhshani of Balochistan using 11 FAO recommended microsatellites markers. Blood samples of unrelated true representative animals of sheep breeds were selected from their breeding tracts and from different Government Livestock Farms in Balochistan province. DNA was extracted with the standard protocol and amplification of DNA done with selected markers in Molecular Biology and Genomics Laboratory in the Institute of Biochemistry and Biotechnology. PCR products were examined on non denaturing Polyacralamide Gel Electrophoresis (PAGE). Genotyping results vanalyzed through the software POPGENE VERSION 1.31 and "POWER STATE" for calculating the observed and expected number of alleles, expected and observed heterozygosity, homozygosity, F-statistics (FST, FIT, FIS), Polymorphic Information Content (PIC), matching probability power of discrimination and power of exclusion. This work provided the genetic data which is useful in breed identification and making effective breeding policies and conservational activities in future according to FAO global Farm Animal Genetic resource data. Average observed heterozygosity, average observed homozygosity, observed number of allels (na) and expected number of alleles for all loci and population in this study were 0.6055, 0.3945, 6.3636 and 4.2805 respectively. Almost all of the microsatellite markers showed significant variations in both breeds of sheep. This work provided the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in future according to FAO global Farm Animal Genetic resource data. Moreover this study can become the basis for further research investigations in sheep breeds in Balochistan and Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1516,T] (1).



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