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Dna Fingerprinting Of Pakistani Buffalo Breeds (Nili-Ravi, Kundi) Using Microsatellite And Cytochrome B Gene

by Rashid Saif | Prof.Dr.Masroor Elahi Babar | Mr. Asif | Prof. Dr. Abu Saeed Hashmi.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Customarily, classification of breed was based on phenotypic traits. In some cases, recent genetic studies have found differences in the structure proposed. In buffalo ,one has to bear in mind that morphological changes were not the result of adaptation to the environment, but have a social significance thus may not be indicative of the genetic relationship. In recent years microsatellites have proved to be very useful for the determination of genetic relationship among population. Comparative studies beiween microsatellite and protein markers have highlighted the advantages of the former. The water buffalo (Bubalus bubalis) holds tremendous potential in livestock sector in many Asian countries, particularly in Pakistan but the genetic data of different buffalo breeds like Nili-Ravi and Kundi is lacking, which need to be established for their genetic identification. Blood samples of unrelated true representative of both breeds (Nili-Ravi and Kundi) were collected from different government livestock farms in Punjab and Sindh respectively. DNA was extracted by inorganic method and amplification of the mitochondrial Cytb gene and microsatellite was done with especially designed primers in Molecular Biology and Genomics Laboratory at Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore. Cytochrome b gene markers have been proved as an efficient and powerful tool for breed characterization and species identification of buffaloes. Several panels of microsatellite markers have also been reported for this purpose. In this study, a panel of nine microsatellite markers has highly Polymorphism Information Content (PlC) were selected, Specific primers were designed for these microsatellite and Cytb gene partial amplification using primer3 software. Then primers were optimized for successful amplification with minimum reagent concentration. PCRs were performed for amplification of these microsatellite and Cytb markers on each sample, Genotyping and sequencing was conducted on all amplicons to find out the different SNP to design haplotypes with the help of bioinformatics software e.g. Blast 2sequence and Chrornas Lite, Further statistical analysis was done by the help of some other software e.g. Popgene version 1.31, Power Stat., Genetic diversity, Allele frequencies, observed and expected homozygosity and heterozygosity, Hardy Weinberg equilibrium, F-Statistics and Gene Flow for all Loci, population's dendogram, Nei's genetic identity and genetic distance! diversity was calculated. The results obtained from this study can contribute to the establishment of routine DNA typing services, beneficial for the buffalo industry as well as in animal forensics for litigation and expedite the police investigation services in Pakistan, which will also be useful for breed characterization and phylogenetic study of aforementioned breeds of buffalo. Availability: Items available for loan: UVAS Library [Call number: 1183,T] (1).

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Sequence Analysis Of Mitochondrial Atpase 8/6 Gene Variants In Sindh Ibex (Capra Aegagrus Blythi)

by Javeria Zafar (2014-VA-222) | Dr. Asif Nadeem | Dr. Maryam Javed | Prof. Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: ATPase 8/6 gene plays a vital role in survival of an organism by generating energy in the form of ATP synthase. Considering the importance of ATPase8/6 gene in energy generating, present study has been designed to characterize this gene in Sindh ibex. The characterization of ATPase8/6 gene might be helpful for deriving phylogenetic relationship among different species and identifying new functions among the related species. Tissue/blood samples (n=15) were collected from Kirthar National Park, Sindh. Standard DNA extraction method was used for DNA extraction. PCR primers were designed by Primer3 software and amplification of gene was done by Polymerase Chain Reaction. PCR product was sequenced bi-directionally by Big Dye TM Terminator on ABI 3130XL Genetic Analyzer. Multiple sequence alignment was performed for polymorphism identification. Genetic diversity was calculated by using DNAsp. Phylogenetic analysis using the MEGA6 software package and an equally weighted maximum parsimony analysis was performed using the close-neighbor-interchange algorithm. The results indicated that Sindh ibex ATPase8/6 gene was highly similar to Capra caucasica. The results of this data might be helpful in designing effective conservation strategies of different species of wild animal. Availability: Items available for loan: UVAS Library [Call number: 2587-T] (1).

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Effect Of Camel Milk Lactoferrin Against Carbon Tetrachloride Induced Hepatic Toxicity In Sprague Dawely Rats

by Nasreen Asghar (2014-VA-546) | Dr. Muhammad Nasir | Dr. Sanaullah Iqbal | Prof. Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Liver is a vital organ involved in regulation of several significant metabolic functions and is responsible for maintaining homeostasis of the body as well as detoxification of chemicals, drugs and other metabolites Chronic Hepatitis and mortality due to liver cirrhosis and hepatocellular carcinoma is common in Pakistan. Camel milk Lactoferrin has antiviral, anti-inflammatory, anti-carcinogenic properties. Liver cirrhosis is a serious and irreversible disease. it is common in Pakistan population which leads to mortality (Khan AA, 1995). (Anderson and Smith, 2001).These problems can be controlled by taking special measures. Nutraceutical foods like camel milk have many beneficial uses in this regard. Camel milk lactoferrin might be used for the cure of hepatic fibrosis induced by CCI4 in Sprague Dawely rats. Seventy five (75) male Sprague –Dawley rats were purchased from National Institute of Health Islamabad kept in animal house of UVAS Lahore and randomly divided into 5 groups under completely randomize design (CRD). In group (2-5) carbon tetrachloride (CC14) was subcutaneously injected with a mixture of 40% CCI4 (a mixture of pure CC14 and sterile olive oil) at 200 uL/100g body weight as single dose, 48 hrs before the starting of treatment or 0 day. After 48 hrs, rats were considered hepatic injured except the (+ve) control group. Among all groups, four groups –ve control, 30mg/kg/b.wt, 60mg/kg/b.wt and 90mg/kg/b.wt were supplied with standard diet plus Lactoferrin (in different concentrations and doses) orally, while control group were provided only standard diet throughout the efficacy study (30 days). Daily feed and water intake was monitored and cages were cleaned regularly. Body weight was recorded before decapitation throughout the experimental period. The efficacy studies data was analyzed through analysis of variance (ANOVA).Statistical significance was defined as P≤0.05.Means were compared for significance difference using Duncan‟s Multiple Range test (DMRt) and Least significance difference test (LSDt). Organ to body weight ratio and decreasing tendency in (-ve) control group was observed, while increasing tendency for body weight was observed in camel milk lactoferrin treatments groups during the study. In the present investigations, higher amount of transaminases (AST,ALT) and cholestatic liver enzyme (ALP) were observed however, their concentrations were significantly decreased significantly in camel milk lactoferrin treatments as compared to rising trend in (-ve) control group. Hypercholesterolemia is due to of oxidative stress induced by CCl4 and characterized by elevated levels of cholesterol .In the present research, it is concluded that camel milk lactoferrin treatments were effective in improving lipid profile.it is also obvious from the current results that camel milk lactoferrin treatments improved the hemoglobin (Hb) level although the effect was found non-significant .Likewise , results of present study also suggested that different treatments, time interval and their interaction had non-significant effects on white blood cells count in rats. Histopathology results of present study given exposed that CCl4 consequences prominent hepatic steatosis, hepatic cord rupture and necrosis .Post treatment of camel milk lactoferrin reduced the severity of CCl4 –induced liver intoxication. Fatty change and necrosis were improved in the histological sections of camel milk post-treated rats.(-ve) control group treated with CCl4 and basal diet showed severe hepatotoxicity, severe inflammation of hepatocytes and portal vein congestion, while group C and D treated with CCl4 ,basal diet and 30mg/kg/b.wt ,60 mg/kg/b.wt camel milk lactoferrin respectively showed moderate improvement in hepatotoxicity .group E treated with CCl4 and 90 mg/kg/b.wt lactoferrin showed mild hepatotoxicity in rats. Availability: Items available for loan: UVAS Library [Call number: 2943-T] (1).

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