1.
Parentage Analysis And Breed Characterization Of Dogs By Microsatellite Markers
by Muhammad Sajid Tahir | 5/31/2010 | Mr.Tanveer Hussain | Mrs.Saeeda.
Material type: ; Format:
print
Publisher: 1990Dissertation note: Pakistan has vast population of dogs belonging to different breeds. Most of the dogs have no pedigree record which is a great threat to conservation of different breeds. No study on DNA fingerprinting of dogs has been conducted in Pakistan. DNA fingerprinting of dogs is necessary to overcome the problems like forensic cases, sale & purchase, individual identity in case of fertilization by more than one male and ownership disputes. Microsatellite markers have been proved as an efficient and powerful tool for parentage testing and breed characterization of dogs. In this study, a panel of microsatellite markers, having high polymorphism information content (PlC) values, was developed. Blood samples were taken from cephalic vein of two breeds of dogs (German shepherd and Labrador retriever). DNA was extracted by Inorganic method. Primers of microsatellite markers were optimized for successful amplification conditions in the Bio-Rad thermocycler. Multiplex PCR was performed, for amplification of these microsatellite markers on 46 samples belonging to 20 families. Genotyping analysis was performed for the PCR products of microsatellite markers on non denaturing polyacrylamide gel. These results were analyzed statistically software "POPGENE 3.3 and POWER STAT". Allele frequency, heterozygosity, homozygosity, polymorphism information content (PlC), power of discrimination and power of exclusion of all microsatellite markers were calculated. Average power of discrimination among non parents, average hetrozygosity, average observed homozygosity and average polymorphism information content (PlC) value for all alleles was 0.809, 0.6345, 0.2913 and 0.724 respectively. Moreover combined power of exclusion reached a significant value of 0.9998. Almost all of the microsatellite markers showed significant variations in both German shepherd and Labrador retriever breeds. Microsatellite "REN41D2Ob" showed maximum variation i.e. 17 alleles and microsatellite"REN49F22b" showed the least variation among all microsatellite markers i.e. 4 alleles. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between German shepherd and Labrador retriever breeds.
Results of this study lead to development of a panel of microsatellite markers which can be used for parentage analysis and breed characterization of dogs. This was a preliminary study on dogs in Pakistan. This facility can be provided on commercial basis to pet owners and kennel clubs. Moreover this study can become the basis for further research investigations in canines in Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 1090,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: ; Format:
print
; Nature of contents: ; 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: ; 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.
Identification And Molecular Characterization Of Shiga Toxin Producing
by Khawar Ali Shahzad | Prof.Dr.Khushi Muhammad | Dr.Tahir Yaqub | Mr.Tanveer Hussain | FVS.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2011Dissertation note: Escherichia coli is normal inhabitant of all the animals and human beings. Its non-sorbitol fermenting biotype (SNF) was detectable in buffalo (90 percent), cattle (80 percent) or rarely in sheep (20 percent) and goat (30 percent). However, SNF E. coli were un-detectable in droppings of rural chickens and feces of donkeys. The SNF E. coli was detected in 100, 92, 71 and 80 percent of the market milk samples and 100, 83, 83 and 53 percent beef samples from Multan, Sandha, Wagha and Sheikhupura Roads, of Lahore city, respectively. However, SNF E. coli was not detected from freshly aseptically collected milk and beef samples but was detectable in over all 96 percent of market raw milk and 82 percent market beef samples. White colored colonies of each positive sample on Sorbitol MacConkey's Agar (SMA) contained gram negative, rods with round ends non spore former. Each of the isolates was sorbitol non fermenter, lactose fermenter, indole positive, Methyle Red positive, Voges Prauskaur negative and citrate negative. However, each of such isolates showed green metallic sheen on Eosin Methylene Blue (EMB) agar.
Each of the isolate was further characterized using polymerase chain reaction (PCR). Bacterial DNA was extracted easily by boiling method when the isolates were grown on Sorbitol MacConkey's agar or EMB agar at 370C for 12-24 hours. The DNA was recovered when the culture was boiled for 5-10 minutes. The DNA of each sample remained stable on storage at 40C for 48 hours or at -200C for 7 days. The isolated DNA (100 samples) when amplified using universal, Stx1, Stx2 and O157 specific primers showed that 82 percent samples were positive for universal primers, 50 percent for O157, 60 % for Stx1 and 51 percent for Stx2. The filtrate of each isolate when diluted as 1:10 dilution and sterilized by filtration induced cyto-pathogenic effect (CPE) on Vero cell line.
It is concluded that SNF E. coli O157 normally exists in intestinal tract of buffalo, cattle, sheep and goat. Counts of SNF E. coli O157 were higher in milk samples as compared to beef samples. More than 80 percent samples of milk or beef were contaminated with SNF E. coli O157. Feces of the animals are presumably main source of SNF E. coli contamination of raw milk and beef. PCR is a quick, reliable, and sensitive technique for confirmation of SNF E. coli O157 in the samples.
Availability: Items available for loan: UVAS Library [Call number: 1221,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: ; Format:
print
; Nature of contents: ; 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.
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: ; 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).
7.
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: ; 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).
