1.
Genetic Characterization Of Pakistani Buffalo Breeds By Mitochondrial D-Loop And Microsatellite Analyses
by Tanveer Hussain | Prof.Dr.Masroor Elahi Babar | Dr. Khalid Javed | Prof. Dr. Irshad Hussain.
Material type: ; Format:
print
Publisher: 2008Dissertation note: Pakistan has various dairy breeds of buffalo and cattle, but the genetic data of different buffalo breeds like Nih, Ravi, Nihi-Ravi, Kundi and Azakheli is lacking which need to be established for their genetic characterization. Blood samples of unrelated true representatives of all breeds were collected from their respective home tracts i.e Nih Ravi (LPRI Bahadarnagar, Okara, BRI Pattoki, Rakh Dera Chahi, Lahore); Nih (Pakpatan,
Minchnabad, Arifwala, Hasilpur); Ravi (Kamahia, Tandlianwala); Kundi (Tandojam, Tando Muhammad Khan, Dadu) and Azakheli (Directorate of Livestock Research & Development Station Surezai, Peshawar and Matta, Swat). DNA was extracted with the use of standard protocol and amplification of the mitochondrial D-loop region was done with specific primers in Molecular Cytogenetics and Genomics Laboratory in the department of Livestock Production. Sequencing of amplified portion of mt DNA D-loop was done. Sequences were analyzed with the help of software blast2sequence. Single Nucleotide Polymorphisms (SNPs) were identified and comparison of 52 mitochondrial DNA haplotypes of all buffalo breeds was done. Genetic distance and identity between five buffalo breeds were calculated and phylogenetic tree was constructed using BioEdit and MEGA 4.1 softwares showing the relationships between different haplotypes. Domestication events were also observed through network analysis. For further confirmation of the genetic structure of buffalo breeds 8 dye labeled microsatehhite markers (recommended by ISAG) were used and genotyping was done. Results were analyzed with the help of different softwares. Genetic diversity, Allele frequencies, observed and expected homozygosity and heterozygosity, Hardy Weinberg equilibrium, F-Statistics and Gene Flow for all Loci, population's dendogram, Neis genetic identity and genetic distance/ diversity was calculated. This work provided the genetic data which is very helpful for determining the genetic diversity of buffalo population, breed identification, animal forensic and paternity cases and making effective breeding policies and conservational activities in future.
Availability: Items available for loan: UVAS Library [Call number: 1114,T] (1).
2.
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: ; 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).
3.
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).
4.
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).
5.
Genetic Study Of Ushic/Dfnb18 By Linkage Analysis
by Muneer Ahmad | Mr. Tanveer Hussain | Prof. Dr. Masroor Elahi Babar.
Material type: ; 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).
6.
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).
7.
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: ; 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).
8.
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).
9.
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: ; 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).
10.
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).
11.
Develoopment Of A Reliable Microsatellites Maarkers Panel For Parentage Analysis In Cattle Breeds Of Pakistan and Its Validatio Through Cytochrome B Gene Sequencing
by Tanveer Hussain | Prof. Dr. Masroor Ellahi Babar | Dr. Ahmad Ali | Dr. Muhammad Wasim.
Material type: ; Format:
print
Publisher: 2013Dissertation note: Pakistan posseses enormous Animal Genetic Resource (AnGR) with 36.9 millions of cattle population. The data on genetic fabric of these breed is yet to be documented for their genetic characterization and identification. This work reports first country wide microsatellite markers and cytochrome b gene based genetic characterization of 10 famous cattle breeds of Pakistan. A total of 352 blood samples from unrelated and phenotypically representative of ten native cattle breeds including Bos indicus; Sahiwal, Cholistani, Red Sindhi, Tharparker, Dhanni, Dajal, Lohai, Bhagnari, Achai and Bos indicus x Bos taurus; Nari Master, and an exotic Bos taurus; Holstein Friesian breeds were collected from their respective home tracts, institutional herds and private livestock farms located throughtout the country. These samples were subject to DNA extraction using inorganic method caliberated to same concentration in Molecular Biology and Genomics Laboratory of the Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore Pakistan. A total of 21 microsatellite markers recommended by the programme for the global management of genetic resources (MoDAD) for breed characterization of Food and Agriculture Organization (FAO) of the United Nations and International Society for Animal Genetics (ISAG) were applied. Multiplex PCR were optimized for amplification and were genotyped using ABI Genetic Analyzer 3130 xl using LIZ as size standard. Genotyping results were analyzed using POPGENE and Arlequin ver 3.5 software. The observed and effective number of alleles ranged from 10 (INRA32) to 43 (TGLA126) and 2.3574 (CSSM66) to 15.0019 (BM6526) respectively in all breeds? The observed and expected heterozygosity estimates ranged from 0.0638 (INRA32) to 0.7101 (BM2113) and 0.6510 (INRA32) to 0.9347 (BM6526) respectively in the experimental samples. Mean values for observed and expected heterozygosity was 0.4943 ± 0.1647 and 0.8164 ± 0.0930 respectively. Mean values for Fis, Fit and Fst in all cattle breeds were calculated as 0.2819, 0.3864 and 0.1456 respectively. Average polymorphic information content (PIC) of all microsatellite loci was 0.81 indicating a high degree of informativeness of all microsatellite markers used. It implies that the same set of markers is equally good and could reliably be used for parentage confirmation in Pakistani cattle breeds. The data produced, also showed least degree of genetic difference between Red Sindhi and Tharparker breeds. This may due to mixing of the two breeds for being in close proximity of their home tracts.
Fragment mitochondrial cytochrome b gene was also amplified using specific primers through PCR of 130 individuals representing all selected breeds and sequencing was done using ABI Genetic Analyzer 3130 xl. The sequences were aligned and analyzed with CodonCode Alligner 4.0.4 software. The analysis revealed highly degree of sequence conservation in all the Pakistani cattle while documenting changes in only 9 nucleotides from 26 individuals whereas multiple nucleotide changes in 5 locations were shown by more than one individual in the data presented. One polymorphic site was found in nucleotide 318 (T?C) in several breeds of indicine cattle while 2 Lohani and 5 Nari Master individuals showed nucleotide changes specific to taurine cattle. Of all the changes found, only three of them caused changes in the amino acid sequence. The UPGMA tree using MEGA 5.1 showed a clear differentiation between taurine and indicine cattle, except for Nari Master Pakistani cattle showing mitochondrial taurine sequences because it's a cross between Bhagnari (Bos indicus) and Australian Draught Master (Bos taurrus). The estimates of divergence among breeds were also low for most breed pairs, except for Nari Master and Dhanni whereas the overall divergence within Bos indicus or within Bos taurus were also very low (0.002 and 0.003, respectively) but the differences between Bos indicus and Bos taurus were significantly higher (0.014) as should be the case.
These results of microsatellite markers have produced a set of information that can be recommended as a reliable marker panel for studies on genetic diversity analysis, parentage confirmation. The cytochrome b data on the other hand not only substantiated genetic diversity analyses but it also proved to be equally good for comparative Phylogenetic analysis of Pakistani cattle breeds and exotic breeds. This work provides most authenticated data and adds a great deal, to already existing information on Pakistani AnGR. This information coupled with prospective data using next generation genetic technologies will assist designing breed improvement focused breeding policies and conservation activities in future.
Availability: Items available for loan: UVAS Library [Call number: 1597,T] (1).
12.
Isolation, Identification And Control Of Vancomycin Resistant Staphylococcus Aureus
by Fakhra Liaqat | Dr. Ali Ahmad Sheikh | Dr. Jawad Nazir | Dr. Tanveer Hussain.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Antimicrobial resistance (AMR) is an emerging issue throughout the world. Vancomycin resistant Staphylococcus aureus has been recently reported from many countries including Asian region. The failure of antibiotics diverts the focus of modern science towards the discovery and application of new alternative antimicrobial agents. Herbal plants not only known for their antimicrobial potential but are being used since centuries for the treatment of infections. This study has been conducted to isolate Vancomycin Resistant Staphylococcus aureus from wounds of hospitalized patients and these isolates were not only tested against Linezolid, Moxifloxacin, and Clindamycin antibiotics but also against the ethanolic extracts of Garlic, Mint, Coriander, Turmeric, Kalonji, Cinnamon and Cloves for their antibacterial activity.
Methicillin resistant Staphylococcus aureus were isolated from wounds and resistance to vancomycin was confirmed according to Clinical and Laboratory Standards Institute recommended method. Minimum inhibitory concentrations of selected antibiotics and selected plant extract was determined by broth microdilution method followed by the measurement of minimum bactericidal concentration by culturing on agar plates.
In current study 104 S. aureus isolates were recovered from 150 wound exudates samples. Resistance to methicillin was shown by 49.04% isolates. Final results yielded, 22 vancomycin intermediate and 5 vancomycin resistant S. aureus strains. Vancomycin resistant isolates were tested for susceptibility against selected antibiotics and ethanolic extracts of plants. Almost all the isolates displayed susceptibility to all three antibiotics and the plant extracts. The data was analyzed statistically by chi square test and one way ANOVA using Statistical Package for Social Science (SPSS) 18.0 software. This study was helpful to find out the effective antibiotics against Vancomycin Resistant Staphylococcus aureus. Plant extracts which were found effective are the best alternate to the conventional antibiotics without having any drawback of antibiotic resistance development.
Availability: Items available for loan: UVAS Library [Call number: 1615,T] (1).
13.
Optimization Of Nested-Pcr For Diagnosis Of Toxoplasma Gondii In Lahore Area
by Amna Arshad bajwa | Dr. Wasim Shehzad | Dr. Muhammad | Dr. Tanveer hussain.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1916,T] (1).
14.
Pathological Studies On Caprine Brucellosis (Brucella Melitensis) In Disteict Shangla Khyber Pokhtunkhwa
by Haider hayat | Dr. Muti-ur-Rehman | Dr. Tanveer Hussain | DR.Ishtiaq.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1919,T] (1).
15.
Sequence Analysis Of Mhc Class-Ll Gene,Drb3 Exon 2 In Beetal And Teddy Goat Breeds Of The Punjab
by Hira paracha | Dr. Tanveer hussain | Ms.Faiza | Ms.Maryam javed.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1965,T] (1).
16.
Assesment Of Diacylglycerol-Acyltransferase-1 Gene Polymorphisms In Nili Ravi Buffalo For Milk Production Trait
by Muhammad Amir zaib khan | Dr. Asif Nadeem | Dr. Abu saeed | DR. Tanveer hussain.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1966,T] (1).
17.
Determination Of Residual Contents Of Pesticide Using Chromatographic Techniques In Rice Samples From Different Geograohical Regions of Punjab
by Abubakar imran | Dr. Tanveer hussain | Dr. Asif nadeem | Ms. Shagufta saeed.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2017,T] (1).
18.
Sequence Analysis Of Polymorphisms In Interferon Alpha Beta And Gamma Genes Of Major Histocompatibilty Complex Class
by Atiya yasmeen | Dr. Tanveer hussain | Dr. Ali ahmad | Prof. Tahir yaqub.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2048,T] (1).
19.
Molecular Characterization Of Heat Shock Protein 40/ Dnaja1Gene In Mareecha Camel Breed Of Pakistan
by Ayesha tariq | Dr. Tanveer hussain | Dr. Ali Ahmad awan | Dr. Muti ur.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2049,T] (1).
20.
Allele Frequency Distribution Of 15 Star Loci In Gujjar Cast Of Punjab Pakistan
by Maira shakoor | Dr. Muhammad Yasir zahoor | Dr. Tanveer hussain | Ms. Shagufta.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2077,T] (1).
21.
Production, Purification And Characterization Of Laccase From White Rot Fungus
by Afrah Shafique (2012-VA-577) | Ms. Faiza Masood | Dr. Abu Saeed Hashmi | Dr. Tanveer Hussain.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Laccase (oxidoreductase, EC 1.10.3.2) are blue copper dependent oxidases and the mainligninolytic enzyme produced by white rot fungus. Laccase catalyze the oxidation of large snumbers of phenolic compounds (Kunamneni et al. 2007; Poonkuzhali et al. 2011). These enzymes have a molecular weight 60-90 kDa and consist of 15–30% carbohydrate. Laccases are the earliest and maximum investigated enzymatic systems. Laccase was initially found by Yoshilda in 1883 in the sap of Japanese laquer tree named as Rhusvernicifera. After a while in 1896, Bertrand and Laborde determined that laccase is a fungal enzyme.(Shraddha et al. 2007; Giardina et al. 2010).
Laccases are present extensively in nature, originating from plants, bacteria and fungi (Poonkuzhali and Palvannan 2011). In fungi, laccases are widely distributed in ascomycetes, deuteromycetes and basidiomycetes. The laccase producing fungus include Trametes versicolor, Pleurotus ostreatus, Polyporus, Trametespubescens, Cerrenaunicolour,PhanerochaetechrysosporiumandFunaliatrogiietc (Dwivediet al. 2011). Laccases occur morein fungi, than in the higher plants. Laccases are also present in few bacteria such as S.lavendulae, S.cyaneus, and M.mediterranea(Viswanath et al. 2008; Arias et al. 2003). In vegetables laccases have been recognized in turnips, apples, pears, cabbages, potatoes, beets, asparagus and various other vegetables (Jhadav et al. 2007).
Enzymes are produced by every living organism, however enzyme produced by microbes have various benefits over the enzyme originated from plants and animal origins.Laccases by nature
are important because of its huge diversity of catalytic activities, economical in production and comparatively more stable than other enzymes.The field of biotechnology proposes expanding possibilities for the production of several enzymes from microorganisms. New methods and techniques have been advanced by using enzyme as biocatalysts to produce big added value products like growing food requirements,good quality chemicals and medicines. Moreover enzymes are also utilized for environmental actions and for diagnostic and analytical motives. (Buchholz et al. 2005). Microbial enzymes are used as cost effective and environmentally sensitive substitutes for chemical processing in several industries and bioremediation. Therefore the commercial demand for microbial enzymes is increasing (Radhika et al. 2013).
Fungal laccases have boundless biotechnological functions across the globe like the decolouration and detoxification of industrial effluent, bleaching of pulp, phenolicselimination from wines, in preparation of biosensors in detergents blockindye transfer- functions (Yaver et al. 2001).It is also used in the formation of anticancer drugs, and included in few cosmetics to lessen their toxicity (Couto and Herrera 2006).In recent years, laccase have been skillfully practiced to the field of nanobiotechnology due to its capacity to mobilize electron transfer reactions without further addition of cofactor(Shraddha et al. 2007).
Laccase is ample in several white- rot fungi that are involved in lignin metabolism (Bourbonnais et al. 1997, Leontievskyet al. 1997). Fungal laccases have immense redox potential (up to +800 mV) than bacteria or plant laccases. The action of these laccases seems to be appropriate in nature and also has significant applications in the field biotechnology. These laccases are associated with the deterioration of lignin and also in the elimination of conceivably lethal phenols appear during the breakdown of lignin (Thurston et al. 1994).
The white rot fungus is corporeal in preference to morphological and composes of those fungi that are adequate of degrading lignin, which is a heterogenous polyphenolic compound in huge amount within the lignocellulose wastes(Eaton and Hale. 1993).Theircapability to deteriorate cellulose, hemicellulose, these are the polysaccharides forming the essential part of lingo cellulose is the basic metabolic processbetween the fungi and happen under the span of environmental conditions.The degeneration of lignindoesn’tprovide net energy so it is degraded during the secondary metabolism in order to gain polysaccharides present in lignin and carbohydrate complexes, supplying energy to which the organisms don’t have access(Jeffrics. 1990).The white rot fungi varyingly secrete one or more three extracellular enzyme namely manganese peroxidase, lignin peroxidase and laccase that are fundamental for degradation of lignin, ant they are generally mentioned as lignin modifying enzymes LMEs (Pickard et al. 1999).
Laccase is the subjects of demanding research in the recent years, because of their several properties like extensive substrate relevance, doesn’t required the inclusion of cofactors because they use oxygen as cofactor which is frequently present in the environment (Eugenio et al. 2009). Maximum number of laccases produced by various organisms is excreted as extracellular enzymes and this makes the purification process quite accessible. Laccase commonly display appreciable extent of stability. Due to these properties laccases are ideally applicable in diverse biological processes such as the treatment of industrial effluent, biopulping and biobleaching (Eggert et al. 2006).
The huge potential of laccase requires advancement in its production and, with huge activities and low cost (Herrera et al. 2007). The use of lignocellulosic agricultural waste as substrates is a tradition for the production of enzyme like laccase because it is ligninolytic in nature (Niladeviet al.2011). It is highly crucial to optimize the fermentation parameters for the adequate production of laccase (Revankaret al. 2007). .
The advantages of agro-industrial leftovers for cultivation media is of immense concern as agriculture waste cut down the expenditure of enzyme production and enhance the understanding on energy protection and recycling (Mansuret al. 2003).These agriculture wastes are comparatively economical and also contain ample nutrients such as lignin, cellulose andhemicellulose. These nutrients serve as inducer to energize the production of enzyme (Vassil et al. 2000).Due to these properties these agricultural waste can be used as substrate for the production of ligninolytic enzymes during the process of fermentation.
Laccase can be produced at varying rates by using a wide range of organisms grown on different substrates and by using several methods of fermentation, such as solid state, semisolid state, and submerged (Rodriguez et al. 1999; Boran et al. 2011). However, for effective laccase production, it is very important to use efficient laccase-producing organisms, suitable fermentation methods, and cheap and widespread sources. Accordingly, one of the most suitable approaches for the production of this enzyme is to use the most efficient agricultural wastes for increasing the production of the ligninolytic enzymes (Elisashviliet al. 2008).
Pakistan is an agricultural country and each year manufactures tons of agricultural by products. These agricultural wastes are accessible in markets at a very reasonable price and can be utilized as substrates in fermentation technique (Minussi et al. 2007). Agricultural waste products like rice husk, wheat bran, corn cob, millet husk and cereal huskhave been utilized by various scientists for laccase production (Osma et al. 2011; jhadav et al. 2009).The chemical properties of these agricultural wastes make them important and economical fermentation medium for biotechnological purposes(Giardina et al. 2010).The cellulose and hemicellulose constituents of lignocellulose wastes are widely used by several organisms but lignin, which is the maximum contrary material to microbial degradation, is transformed conveniently by only few organism of thw white rot fungus (Dwivedi et al. 2011).Lignin serves as a barrier that protects cellulose and hemicellulose from enzymatic attack, however, white rot fungi can attack this barrier in order to obtain energy from cellulose. These fungi produce different extracellular ligninolytic enzymes such as laccase, manganese peroxidase, and lignin peroxidase (Couto et al. 2006).
Fermentation is a biological approach that is used for the transformation of complicated substrates into basic composites by different microorganisms like bacteria and fungi. In the procedure of this metabolic breakdown the microorganisms also release various added compounds like carbon dioxide and alcohol asidefrom the conventional products of fermentation. These added compounds are known as secondary metabolites (Pandey et al. 1999). These Secondary metabolites span from enzymes, antibiotics, peptides and growth factors (Balakrishnan and Pandey. 1996; Machado et al. 2004; Robinson et al. 2001). They are also known as bioactive compounds becausethey carry biological activity(Demain et al. 1999).
Submerged fermentation is a type of fermentation in which components are present in a liquid media like broths and syrup. The co-active composites are poured into the fermentation broth. In this media the substrates are employed quiet immediately, due to this reason the nutrients in the media are either fortified or regained continuously. This type of fermentation approach is optimum for microorganisms such as bacteria, fungi because they depend upon on immense moisture content. The increased benefit of this approach is that the purification of the desired products or enzymes is quiet effortless. Submerged fermentation is especially used in the abstraction of secondary metabolites that are utilized in liquid form (Subramaniyam et al. 2012).
Furthermore 75% of the commercial enzymes are made by using submerged fermentation, it also supports the usage of genetically modified organisms to a large expanse then solid state fermentation. Submerged fermentation is also used on large extent because it doesn’t require equipment concerning solid state. On the contrary solid state fermentation is a mechanism operated in absence of free flowing water by utilizing solid support in form of natural substance ( Poonkuzhali et al . 2011). .
The major purpose of conducting this research is to design optimized fermentation process which produces effective amount of enzyme by using agricultural wastes. The use of agricultural wastes as substrates is economical and increase awareness on energy conservation .The enzyme can be used further for bioremediation because it not substrate specific and can act on broad range of substrates.
Availability: Items available for loan: UVAS Library [Call number: 2213,T] (1).
22.
Genetic Effect Of Interferon Gamma On Bovine Resistance Against Mycobecterium Bovis
by Syed Ahmed Raza Rizvi (2012-VA-819) | Dr. Maryam Javed | Dr. Tanveer Hussain | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Bovine tuberculosis is a disease caused by the species included in the Mycobacterium tuberculosis complex. IFN-GAMMA are a family of conserved innate immune recognition receptors that trigger adaptive immune responses. IFN-GAMMA play an important role in host defense against mycobacteria, especially by mediating the response to mycobacterial triacylated lipopeptides.
The objective of this study is the identification of single nucleotide polymorphisms within the coding region of IFN-GAMMA gene to evaluate its potential for enhanced the resistance to bovine tuberculosis in Nili-Ravi buffalo breed. Fifty blood samples of Nili-Ravi breed were collected from UVAS Pattoki Campus, Research Farm B and Buffalo Research Institute (BRI) Pattoki. Inorganic method was used for DNA extraction, for amplification of the coding region of IFN-GAMMA gene PCR (Polymerase Chain Reaction) was used using specially designed primers and the PCR products were sequenced through Sanger’s Chain Termination method. For the analysis and alignment of sequencing the results obtained after sequencing were analyzed and aligned using the CLUSTAL W and BLAST software. After all these analysis Ten SNPs were identified in the coding region of IFNG mentioned in table. The Eight SNPs identified in the coding region of INTERFERON GAMMA were in this order P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, and P8 C >T. The one SNP found in the current research is in compliance with the (Sun et al. 2012) research on INTERFERON GAMMA hence Nine SNPs found in the current research are novel in Nili Ravi buffalo. Research on IFN-GAMMA hence Seven SNPs found in the current research are novel in Nili Ravi buffalo. The SNPs in the exonic region that is P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, P8 C >T, among them four were transitions and four were transversion .
Population genetic analysis and allelic distribution at all loci was analyzed using
Summary
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POPGENE 32 software indicated that at [P3=0.354539>0.05] , [P5=0.365524>0.05]followed the assumptions of the Hardy-Weinberg equilibrium indicating that the alleles were randomly distributed throughout the population, no migration had occurred, no bottlenecks happened and population remained large in numbers. This Non-significant and obeying HWE, so can be potential marker for genetic selection. At [P1= 0.000032< 0.05], [P2=0.038766< 0.05] and [P7=000394< 0.05] the probability value below 0.05 indicated that population at these polymorphic sites was not obeying Hardy-Weinberg equilibrium. This indicated that at these positions alleles were not equally distributed in population. It can be concluded from my research that the SNPs identified in the current research may also hold potential for marker-assisted breeding programs, with the aim of breeding more BTB-resistant animals and herds within both the national farms and the private sector. Availability: Items available for loan: UVAS Library [Call number: 2419-T] (1).
