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1. Comparative Studies On The Sensitivity Of Polymerase Chain Reaction (Pcr) And Microscopic Examination For The Detection of Trypanosoma Evansi in Horses

by Muhammad Asif Muieed | Prof. Dr. Zafar Iqbal Chaudhary | Dr. Azhar Maqbool | Mr. Asim Aslam | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: The polymerase chain reaction (PCR) was standardized and its efficacy was evaluated against microscopic examination i.e. Giemsa stained smear method ['or the diagnosis of Trypanosoma evansi infection (Surra) in horses. l3lood samples were collected from 100 suspected horses from different localities in Lahore. Under aseptic precautions blood smears were prepared, after drying and fixing with methanol, slides were stained by Giemsa stain method of staining. By stained blood smear method 5 out of 100 horses were found positive For T. evansi infection. The polymerase chain reaction (PCR) was carried out on the blood of' the same suspected horses to evaluate its efficacy in the diagnosis of' T. evansi infection and to compare its diagnostic value against the microscopic examination method currently in use. For this purpose total genomic DNA was extracted from suspected blood samples. The PCR reaction was performed in a 50tl reaction mixture containing I X Taq BuFfer, 0.2 mM dNTP Mixture. I .5 mM MgCIl2 2.5 U/1i1 Taq Polymerase. 4uM of' each primer, 2 ul of DNA extracted and 31.5 p1 of DNase - free deionised water. The tubes containing the mixture were subjected to 30 cycles of amplification in a thermocycler. During each cycle the sample of' DNA was denatured at 93° C' For 30 seconds, annealed at 45° C For 30 seconds and extended at 720 C For I minute. Prior to the cycling and at the end of' cycling the mixture was subjected to incubation at 93° C for a period of 3 minutes and final extension at 72° C for a period of 5 minutes, respectively. PCR product was then characterized by 2.5% of agarose gel electrophoresis. To confirm the presence of DNA and to estimate its size it was compared with a DNA ladder and was photographed with a Polaroid camera. The polymerase chain reaction (PCR) revealed 16 positive cases out of 100 above mentioned suspected cases. These 16 positive cases diagnosed by polymerase chain reaction (PCR) also included animals, which were diagnosed by stained blood smear method. It can be concluded that polymerase chain reaction (PCR) is a superior and sensitive (16%) in comparison with the microscopic examination i.e. Giemsa stained smear method (5%). Polymerase chain reaction (PCR) is more effective in cases where the parasitemia is low and this test could be used in other species of animals especially camels where the disease is more chronic and difficult to confirm by. other routine methods. PCR would not only ensure early diagnosis and treatment in individual animals but can detect animal reservoirs of infection and would help to eliminate threat to equine and camel herds which are grazed and housed together and where blood sucking mechanical fly vectors are ever present. Availability: Items available for loan: UVAS Library [Call number: 0860,T] (1).

2. Molecular Investigation Of Mental Retardation Locus (Mrti)/Gene Prss12 By Linkage Analysis

by Zafar Ali | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Muhammad Asif.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Mental retardation (MR) is a condition in which a person having an intelligence quotient (IQ) lowers than 70. It is also associated with a deficit in adaptive behavior such as communication and daily living skills. Mental retardation is either non-syndromic or syndromic. It is one of the most common genetic disorders and it affects about 1-3% of the human population, with a ratio of males higher than females. The present study was can-ied out to determine the prevalence of families having mental retardation in Pakistani population. In the present study, 7 MR families with three or more affected individuals with MR were enrolled. Family history was taken and pedigree was made personally by visiting the families. The blood samples were collected from the enrolled families. Then DNA was extracted from the blood samples collected from these families by standard inorganic protocol. After isolation of DNA from blood samples, 3 STR markers (D4S191, D4S2392 and D4S3024) in vicinity of mental retardation (MR) locus (MRT1)/gene PRSS12 were amplified on each sample of each family by PCR. The amplified PCR product was first checked on agarose gel and then genotyping analysis (linkage analysis) was performed on non denaturing polyacrylamide gel (PAGE). After polyacrylamide gel electrophoresis, picture of the gel was taken and alleles were read manually with larger allele donated by 2 and smaller by 1. After that haplotype was constructed to determined the pattern of inheritance among the affected and normal individuals of each family under study and also to determined that a family was linked or unlinked to mental retardation locus (MRTI)/gene PRSS12. None of the family was linked to mental retardation locus/gene PRSS12. The families which remain unlinked to the reported loci during screening signifies extreme genetic heterogeneity of MR which is not surprising because about 50% of human protein coding genes are expressed in the brain and it provides an excellent resource material for mapping of the new genes which will shed light on the complex pathways involved in the development of learning and memory in those population. The pedigree of each family in the present study showed that most of the marriages are cousin marriages; therefore this study may play a role in creating awareness about the effect of cousin marriages that is the first step towards decreasing socio-economic burden of the country by genetic counseling and also to prevent mental retardation in Pakistan due to inbreeding. Mental retardation locus (MRT1)/gene PRSS12 was studied for linkage analysis in seven families from different areas of District Swat and Peshawar of Khyber Pakhtunkhwa province of Pakistan. None Out of seven families was linked to mental retardation locus (MRT1 )/gene PRSS 12. All the seven families remain unlinked to this locus. It is concluded that Mental retardation is a complex genetic disorder and needs further studies to identify the already known locus or to explore novel loci through genome wide scan responsible for mental retardation in these population. This will provide opportunities of genetic counseling to these populations and will ultimately result in prevention of mental retardation in Pakistani population. Availability: Items available for loan: UVAS Library [Call number: 1171,T] (1).

3. Status Of Wild Life Close To Indian Border Area At Ravi Siphon, Pakistan

by Muhammad Asif Munir | Dr. Zulfiqar Ali | Dr. Muhammad Mahmood-ul-Hassan.

Material type: book Book; Format: print Publisher: 2010Dissertation note: The proposed study was carried out at Ravi Siphon area in Sheikhupura district near Lahore, Punjab, which is an important site along the Indian Border. In this study, main emphasis was given on the population dynamics of birds, mammals, amphibians and reptiles in relation to the wetland characteristics particularly different types of riverine habitats, different seasons (summer, winter), relationship between different species of animals, seasonal changes, species identification, census of biodiversity and their population assessment. In addition, natural flora of the site was also recorded. Wildlife is an excellent indicator of the overall health of an ecosystem. On this research identification of nature and severity of problems being faced by biodiversity was identified and concluded as recommendation for the welfare of our wildlife to declare the site as a protected area. Waterbirds depend on Ravi Siphon wetland for a variety of activities which include feeding, breeding, nesting and moulting. The highest number of waterbirds is often found in wetlands which have the greatest diversity of plant species and vegetation types, or where there is permanent water. In hot summer wetlands become visiting places for summer visitor birds and they provide a drought- refuge for several species of water birds. During the study there were 87 species of birds recorded. According to seasonal distribution 24 winter visitors, 54 residents, 6 summer breeder and 3 year round visitor were recorded. In the study area, some birds were seen in a large diversity and others were seen rarely. The present study about the status of bird species showed that 1 very common, 9 common, 20 fairly common, 34 uncommon, 12 rare and 11 very rare bird species were recorded. Monthly data was taken during the whole year (May 2009-April10) and 772 total birds population were found. In May 319, Jun 354, July 375, August 390, September 316, October 432, November 349, December 395, January 373. , February 389, March 401 and in April 363 bird's population were recorded. Relative abundance, Census Index and Shannon Weiner Diversity Index were also calculated for studied bird data. The most dominant bird of the area was Indian Cliff Swallow found having relative abundance 5.69. The other dominant birds were Little Green Bee-eater (3.4), Large Egret (3.96), House Sparrow (2.3), and Bank Myna (2.6). Census Index was found to be 1.64 and Shannon Weiner Diversity Index was 5.98 found. Wildlife other than birds was also recorded. Three species of amphibians, eight species of reptiles and significant species of mammals were studied. A great variety of plant trees were also found at Ravi Siphon. Availability: Items available for loan: UVAS Library [Call number: 1173,T] (1).

4. The Study Of Gene Gjb2/Dfnb1 Causing Deafness In Humans By Linkage Analysis From District Peshawar

by Noor Badshah | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Muhammad Asif.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Hearing impairment is the partial or complete inability to hear that leads to compromise the development of normal language skills. Among all the sensory impairments in humans, hearing impairment is the most common. It is estimated that at least 50% of the cases are due to genetic factors. Hereditary hearing loss may be syndromic or non-syndromic; about 30% of deafness cases are syndromic, while 70% is non-syndromic. It is estimated that the prevalence of profound bilateral hearing loss is 1.6 per 1000 in Pakistan and 70% of hearing loss arises in consanguineous families. The main pattern of inheritance of deafness in Pakistani population is autosomal recessive and to date more than 145 loci and 26 genes have been identified for non-syndromic recessive deafness. More than 400 disorders associated with hearing loss shows extreme genetic heterogeneity and complexity of the mammalian inner ear. As more genes are identified, the elucidation of the function of the proteins that these genes encode contributes greatly to the understanding of cochlear mechanisms and their role in disease causation. The gene involved, GJB2, encodes the connexin26 molecule. Connexin26 is a component of gap junctions, the links that allow small molecules to pass from one cell to the next, and this protein is found in several places in the body, including the epithelial supporting cells surrounding the sensory ear cells of the cochlea.The sensory ear cells of the cochlea allow potassium ions to pass through their upper surface during normal reception of sound, and these potassium ions must be recycled through the base of the ear cells and the supporting cells and fibrocytes back into the high-potassium endolymph that bathes the tops of the ear cells. The aim of this study was Linkage analysis for DFNB1 locus involved in causing hereditary deafness in families from Khyber Pukhtunkhwa. A total of 10 families were enrolled from different areas of Khyber Pukhtunkhwa province. I have studied 8 families of these 10 (i.e.) family no. 2, 3, 4, 5, 6, 8, 9 and 10. The families have at least three affected individuals. All the families showed recessive mode of inheritance. For linkage analysis studies for DFNB1 locus, three STR markers D13S175, D13S292, and D13S787 were genotyped using Polyacrylamide gel electrophoresis (PAGE) and haplotypes were constructed to determined, linkage with DFNB1 locus. From a total of 8 families, a single family was linked to DFNB1 locus. The DFNB1 locus is the first non-syndromic deafness locus mapped to chromosome 13q12. Availability: Items available for loan: UVAS Library [Call number: 1191,T] (1).

5. Genetic Diversity And Differentiation Of Domestic Buffalo Of Pakistan Through Sky And Zfy Genes Of Y Chromosome

by Muhammad Mudassar Manzoor | Mr. Taneer Hussain | Mr. Muhammad Asif | Prof. Dr. Abu.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2011Dissertation note: Livestock sector plays a vital role in the economy of Pakistan. Main contribution of milk comes from buffaloes and cows. Water buffalo (Bubalusbubalis) is one of the major elements of livestock in the country and possess great importance for economy in the form of milk and meat productions. Nili, Ravi,Nili-Ravi, Kundi and Azakheli are major breeds of water buffalo to be found in different areas of Pakistan. Conventional classification of breeds 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 Y chromosomal genes have proved to be very useful for the determination of genetic relationship among population. Comparative studies have highlighted the advantages of the SRY and ZFY genes of Y chromosome.These genes have been considered as competent and powerful tool for the purpose of breed characterization and species identification of buffaloes. In livestock sector, water buffalo (Bubalusbubalis) has shown great prospective in numerous Asian countries including Pakistan. Unfortunately, there is lack of genetic data of different buffalo breeds like Nili-ravi and Kundi which needs to be established for their genetic identification.Blood samples from true representative animals of each of the two buffalo breeds (Nili-Ravi and Kundi ) were collected from different Government livestock farms and their respective home tractsin Punjab and Sindh respectively. DNA was extracted by inorganic method and amplification of the SRY and ZFY(exon 5) genes of Y chromosome was done with especially designed primers using Primer3 software in Molecular Biology and Genomics Laboratory at Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore.Specific primers are designed for these genes amplification. Then primers were optimized for successful amplification with minimum reagent concentration. PCR was performed for amplification of SRY and ZFY(exon 5)genes on each sample. Sequencing was conducted on amplicons to find out the different single nucleotide polymorphism (SNP) to make haplotypes with the help of bioinformatics software like Blast 2sequence and Neighbour Joining phylogenetic tree was constructed by using MEGA version 5 (Tamuraet al. 2011). The results obtained from this study now can contribute to the establishment of routine DNA typing service to the advantages of the buffalo in livestock industry. Availability: Items available for loan: UVAS Library [Call number: 1378,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: 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).

7. Molecular Characterization Of Cacna 1H Gene To Fine Out Association Of Polymorhism In Childhood Absence

by Qurrat-ul-Ain | Dr. Muhammad Wasim | Dr. Muhammad Asif | Mrs. Shagufta.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1472,T] (1).

8. Designing Of Oligo Pool All For The Selection Of Superior Dairy Animals In Pakistan

by Kamran Abbas | Prof. Dr. Masroor Ellahi Babar | Dr. Aftab | Mr. Muhammad Asif.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Livestock has an important role in an agriculture based country like Pakistan with a large number of dairy animals. However the average daily milk yield of dairy animals is very low. There is need to improve milk yield by the selection of superior dairy animals using latest genomic selection procedures. Selection of superior animals on the basis of genetic markers has a tremendous potential for breed improvement across the globe. Substantial advances have been made over the past decades through the application of molecular genetics used in industry programs for several decades and is growing, the extent of use has not lived up to initial expectations. Most applications to date have been integrated in existing programs on temporary basis. Among various molecular markers the Single Nucleotide Polymorphism (SNP) is one of the major genetic marker used worldwide. Through SNP genotyping selection of phenotypic superior animals can be done. There are many techniques used for SNP genotyping but the most advanced technique is Veracode GoldenGate Assay by Illumina. Illumina's VeraCode technology with the BeadXpress (BX) Reader is ideal for high-throughput small to mid-scale genotyping studies and SNP validation. BX leverages the power of digital holographic codes and the robust GoldenGate Genotyping Assay to provide a detection method for multiplex assays requiring high precision, accuracy, and speed. A custom assay of 48, 96, 144,192 and up to 384-SNPs OPA (Oligos Pool All) is designed using Illumina's Assay Design Tool and manufactured by Illumina. As a first step for designing of Veracode GoldenGate Assay the development of Oligo Pool All (OPA) is necessary. The OPA was designed by using the genes for milk production, growth, fertility, health and other performance traits. The SNP's in these genes was searched from different gene banks and after proper arrangement the files were sent to Illumina for scoring. After scoring the OPA was finalized for the Veracode GoldenGate Assay for the selection of superior dairy animals in the country using the highly robust BeadXpress technology. The development of OPA for the selection of superior dairy animals was done for the very first of its kind based on modern technology, Veracode GoldenGate Assay in Pakistan. This will greatly help the livestock and dairy development departments, livestock owners, breeders, forensic agencies and researchers to use this unique panel of molecular markers for the selection of superior animals on the basis of marker assisted selection. Availability: Items available for loan: UVAS Library [Call number: 1528,T] (1).

9. A Study On Variable Degrees Of Angles In Z-Plasty Technique To Evaluate Extent Of Relaxation Of Contracted Skin

by Muhammad Asif | Prof. Dr. Muhammad Arif Khan | Dr. Muhammad | Dr. Shehla Gul Bukhari.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1648,T] (1).

10. Prevalence, Identification And Pathogenesis Of Clostridium Chauvoei In Cattle And Buffaloes In Punjab

by Muhammad Asif Idress | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Muhammad Younus.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: In the first phase of the project, the sampling of diseased animals presumably affected by Black quarter was carried out from six districts of Punjab belonging to three different zones. Around two hundred and fifty samples from each zone were collected and were subjected to bacterial culturing and isolation procedures followed by biochemical identification mechanism. The prevalence of Black quarter in Cattle and buffaloes were thus calculated for each district and zone. Highest prevalence of BQ in Zone II was observed (27.2%) for cattle while in case of Buffaloes highest prevalence (3.2%) was noted in Zone I. similarly higher Prevalence of BQ was noted in 1st quarter of year for Zone I followed by zone II and III while 2nd quarter of season was showing higher prevalence of BQ in zone II and III. During 2nd phase of experiment tissue samples were inoculated in RCM and blood agar for the re-isolation of C. chauvoei, identified on the basis of colony characteristics and later on subjected to biochemical tests for the confirmation of the isolated organism. Then it was further confirmed through Polymerase chain Reaction for the identification of the causative agent i.e. C. Chauvoei on the basis of 16S rRNA gene sequence. Another set of primers corresponding to alpha toxin gene sequence of C. chauvoeui was also used which strengthened the belief that this strain of C. chauvoei possessed alpha toxin producing ability. During third phase of project blood samples collected were subjected to hematological estimation for buffaloes and cattle having confirmed as BQ This study revealed significant effect on RBC's count and white blood cells count (P<0.05), while Differential leukocyte count were also showing significant different as compared to Non-infected (P< 0.05). Serum samples were tested for the change in levels of different enzymes. It was found that blood-glucose level and ALT levels were not significantly higher (P>0.05) when compared with control values, Values of AST, CPK and LDH were found significantly higher (P< 0.05) in all infected animals. Histopathology of affected muscle tissues of both cattle and buffaloes was done to study microscopic changes in the muscle fibers and surrounding tissues. Lesions were somehow disappointing as compared to the magnitude of gross lesions. There were segmental degeneration, Zenker necrosis, discrete edema, occasional neutrophils and emphysema in affected muscle. Finally, alpha toxin (hemolysin) in culture supernatant of RCM broth was titrated against 2% washed RBC's of cattle, buffalo, sheep, goat, chicken, rabbit and mice to study the hemolytic activity of the toxin. It was found that highest percentage of hemolysis was observed in mice followed by cattle, sheep, buffalo, chicken and rabbit respectively at 25°C. Higher the dilution of toxin, lower the extent of hemolysis. At 37°C variable results were obtained. It showed the biological activity of alpha toxin is also temperature dependant. Availability: Items available for loan: UVAS Library [Call number: 1664,T] (1).

11. Molecular Diagnosis Of Trypanosomiasis In Pet Dogs Of Lahore

by Muhammad Asif (2007-VA-460) | Prof. Dr. Kamran Ashraf | Dr. Nisar Ahmad | Dr. Jawad Nazir.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Trypanosomaevansiis a protozoa that causes surrain a wide variety of mammals. It is widely reported in adult dogs (Rashid et al. 2008; Defontis et al. 2012). Trypanosoma evansiis utmostcommonlyexisting trypanosome in animals. It is a salivarian pathogen (Hoare. 1972). Stomoxyand Tabanidssppare menifested as mainvectors universally. Oral spreadis also reported in both wild and domestic animals (Adams and Lionnet. 1983). Since 2008, surra became obligatory not only in horses, because it has been considered as a multi-species disease by the OIE (OIE. 2008; Salim et al. 2011). Surra usually follows an acute course of infection in dogs, though it is sporadically prevalent (Ravindran et al. 2008). Outbreaks of canine trypanosomiasis have been reported in India, Iran,Brazil, and South America (Herrera et al. 2004; Morteza et al. 2007; Umezawa et al. 2009). Trypanosomaevansionly has been reported from subcontinent (Ravindran et al. 2008). Causative agent for American trypanosomiasis is Trypanosoma cruzi (T. cruzi) and African trypanosomiasis (surra or sleeping sickness) whose causative agent isTrypanosomaevansi(T.evansi).These are two forms of canine trypanosomiasis.It was originally an enzootic disease mingling in mammals and birds, which served as areservoir. The disease became zoonotic due to interaction between rural populations and natural foci, which are the results of biologicalinequity (Johns et al. 2000). Peracute to acute infections due to trypanosome result in high temperature, hemorrhages in the mucosal and serosal sides. Anemic condition of the patient is produced due to loss of RBCsfrom circulationby the mononuclear phagocytic system which is the cardinal feature of trypanosome infection.In chronic infections, anemia may be resolved due to little parasitic load in blood at capricious degrees. (Urquhart et al. 2002). Some note able signs may compriseedema of throat and head, Blindness due corneal opacity, Temperature and anorexia. Larynx may alter the voice of the dog due edema, which can complicate with rabies. Infected dogs are considered as a risk factor in household spread of the Chagas disease in humans (Cohen and Gurtler.2001). In native animals, dogs have the flankingassociation with humans; they may assumedconsiderable epidemiological importance in the perspective of public health and zoonosis. In humans,T.evansiproduce chronic pathological changes, which includes congestive cardiac insufficiency, finding of which isproblematic and can be unexploited due to multisystemic nature of the infection; it increases the need for epidemiological and experimental support. Moreover, causative agent of trypanosomiasis must beconfirmed by laboratory analysis, which can make availablesignificantprovisionwhen using suitable techniques, suitable reagents, and subsequent good laboratory practices (Eloy & Lucheis. 2009). There has been development of several compounds with value against canine trypanosomiasis, however none of these products have been produced in a large commercial scale or even accessible in the market. The apparent inaccessibility of new trypanocides in the market have continued a great challenge to the treatment of the infection. Diminazine aceturate dose of 3.5 mg/kg in T.congolense infection; 7 mg/kg inT. brucei andT. evansi(Aquinos. 2007) has shown efficacy when used to treat canine trypanosomiasis.However, treatment does not provide satisfactoryresults but only sustained the life of the dog for some reasonable period (Amora. 2004). Dogs were vaccinated with a fixed T.rangeli against canine trypanosomiasis recently (Basso et al. 2007). Experimental infections of the vaccinated dog produced disease of low parasitaemia apparently from vaccine induced immunity. Furthermore, feeding of the vaccinated dogs with the nymph stage of triatomine reduced the rate of infection in the bugs. Since dogs are the reservoir of Chagas disease in man, advances in this area could reduce the rate of infection of kissing bug which will in turn aid in the control of the disease in man (Basso et al. 2007) are necessary to establish the diagnosis.Sensitivity of direct parasitological examination is directly related to parasitic burden, biological material. The diagnosis oftrypanosomiasis is based on combination of comprehensive clinical inspection, appropriate sample collection, sample size, suitable diagnostic tests and suitable conduction of tests and logical interpretation of results. In canine trypanosomiasis where disease prevalence is great, some tests of low diagnostic sensitivity may suffice (OIE. 2008). Parasitological diagnosis could be made by microscopic inspection either of blood, lymph node or CSF of infected dogs (François et al. 2005). Pet dogs have been the companion of human being since ages, and shares the environment and belongings. Trypanosoma is found in dogs causing health problems effecting their routine activities. Rapid and accurate diagnosis of the infection is must to do for better therapeutic approach and early recovery of the animal. PCR is gold standard test for the molecular diagnosis of disease leading to quick diagnosis, early recovery and cost saving. So, regarding to disease importance and dogs domestications which is increasing day by day in and around Lahore area, we have focused this species to determine the Trypanosoma evansistatus in dogs in this area. This whole study is based on two diagnostic techniques i.e. screening through microscopic examination and confirmation of these samples via PCR with details regarding age, sex and breed association with the disease. Availability: Items available for loan: UVAS Library [Call number: 2259-T] (1).

12. Clinico-Pathological And Pathomorphological Studies On Co-Infection Of Avian Influenza (H9n2) With Escherichia Coli In Broiler Chicken

by Shahid Jaleel (2003-VA-93) | Dr. Muhammad Asif Idrees | Prof. Dr. Muhammad Younus | Dr. Muhammad Arshad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: E.coli is an important pathogen of domestic poultry and is prevalent in commercial poultry. LPAIV H9N2 infections are emerging respiratory problems in poultry industry, causing huge economic losses especially in the presence of other co-infecting pathogens such as E.coli. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. The aim of the present study was to investigate the infection of LPAIV A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) in chickens challenged with E.coli (O78:K80). This study had three objectives. First, it is designed to develop co-infection experimental models LPAIV (H9N2) + bacteria (E.coli) in the avian model. Second, it aims to study the hematological and biochemical alterations during co-infection in avian model. Finally to study the pathological and histological alterations during co-infection in avian model, this study will help researchers and veterinarians in implementation of necessary control measures. E.coli stockculture was prepared by inoculating MacConkey’s agar with a loop full of reference E.coli strain culture and incubating at 37°C for 24 h. The estimated colony count was confirmed by plating 0.1 ml of a 104 and a 105 dilution of the final culture onto separate MacConkey’s agar plates. Avian influenza A virus, A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) was obtained from Poultry Research Institute (PRI) Rawalpindi Pakistan. Viral stocks were prepared and titrated in 9-day-old to 10-day-old chicken embryonated eggs the median embryo infectious dose (EID50) was computed using previously reported approaches The viral stocks were diluted in medium containing antimicrobials to give a final titre of 106 EID50/ ml The study were ran on 80 broiler chicks (3week old), procured from local hatchery. All fowl were held serologically innocent and free from flu virus by haemagglutination inhibition (HI). Chicken were infected under experimental conditions with E.coli (O78:K80) and low pathogenic avian influenza (LPAI) strain (A/chicken/Pakistan/UDL-01/08) (H9N2) alone or in combination. The experimental groups were identified as follows: negative control, E.coli, AI, and E.coli plus AI. Infected birds showed clinical signs of differing severity, with the most prominent disease signs appearing in birds of the E.coli plus AI group. Moreover, birds in E.coli plus AI group showed significant decrease in weight, enhanced macroscopic and microscopic pathological lesions. Specifically, the survival rate was 60%, 90%, and 100% in birds inoculated with E.coli + AI, E.coli and control negative or AI virus alone, respectively. Hematological studies revealed anemia, thrombocytopenia and leukopenia especially in co-infected birds. Biochemical studies revealed a significant decrease in total protein, glucose and albumin concentration with significant increase of activities of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. Prominent increase in creatinine, urea and uric acid were significantly detected in the infected chicken. The results showed that experimental co-infection of E.coli and H9N2 increased the severity of clinical signs, mortality rate and gross lesions and suggest than E.coli infection can induce higher economic losses and mortality if H9N2 LPAIV is also present. The HI titer against LPAIV infection in the co-infected group was significantly higher than the HI titer of AI group, which may indicate that E.coli could promote the propagation of H9N2 LPAIV or stimulate the immune response. The present study revealed that co-infection E.coli and H9N2 LPAIV caused more serious synergistic pathogenic effects and indicates the role of both pathogens as complicating factors in poultry infections.   Availability: Items available for loan: UVAS Library [Call number: 2552-T] (1).

13. Immunomodulatory Effects Of Feeding Allium Sativum Against Infectious Bursal Disease (IBD) Vaccinated Broiler Birds On IBD Vaccine.

by Uzma Riaz (2008-VA-286) | Prof. Dr. Muhammad Younus | Dr. Muhammad Asif Idrees | Dr. Iahtasham Khan.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: A number of feed additives including antibiotics have been extensively used in poultry diets for the purpose of weight gain to improve feed efficiency and growth rate. However, use of antibiotics has restricted due to the bacterial resistance and the issue of residues which make the chicken meat harmful for human consumption. So the medicinal plants are gaining interest as alternative feed strategies now a day because of their low cost, easy availability and presence of no residues. Garlic was used for the medicinal purposes and as a health supplement by the ancient Egyptians. It is a natural feed additive and is antimicrobial, immune stimulator, antiviral, antifungal, anti-parasitic, antithrombotic, antioxidant, anti-cancerous, and vasodilator activities. Previous studies indicate that it has beneficial effects on the immune system and is a best immune stimulator then the other herbal plants and medicines. Therefore, the present study was designed to estimate the immunomodulatory effect of garlic to commercially available IBD vaccine in enhancing the immune system. Total N=99 day old broiler chicks were purchased and kept in the experimental shed of CVAS Jhang. Birds were divided into three groups A, B, C and group B and C were further divided into three subgroups (B1, B2, B3 and C1, C2, C3). Group A was treated as control group and was administered with commercially available IBD and ND vaccine and routine diet while group B was administered with garlic at the rate of 4%, 5%, and 7% along with vaccine to see the impact of different levels of garlic (Allium sativum) on the immune system and to see the toxic effect (if any) of high dose of garlic. Group C was only administered with garlic in fee Summary 44 at the same rate as to group B. At the end of study birds were slaughtered to check the effects of garlic administration. Positive effect of garlic has been reported by many studies. Garlic is a medicinal herb used for the prevention and treatment of many diseases, because of having antiviral, antibacterial and antifungal activities. Also act as a good growth promoting agent and have beneficial effects on the immune system. Results of the study indicate that administration of garlic powder in different doses alone and combined with commercial IBD vaccine have good effects on the growth, blood parameters and the immune system of the broiler birds. Availability: Items available for loan: UVAS Library [Call number: 2561-T] (1).

14. Active Surveillance Of Avian Influenza In Sentinel Live Bird Markets Of District Rawalpindi

by Muhammad Asif (2015-VA-435) | Dr. Mamoona Chaudhary | Prof. Dr. Mansur-ud-din Ahmed | Dr. Hamad Bin Rashid.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Influenza is a highly contagious, acute illness in humans. Influenza viruses have negative-sense RNA genomes and are placed in the Orthomyxoviridae family grouped into three types A, B and C on the basis of the internal nucleocapsid or the matrix protein. Droplet and airborne are the most common modes of transmission. In Humans infection appears to be direct or indirect exposure to infected live or dead poultry or contaminated environments, such as live bird markets. The outbreak can be subsided by applying biosecurity measures, controlling poultry movement, using inactivated vaccines and initiating an Avian Influenza surveillance network throughout the country. Avian influenza virus is prevalent in live birds markets of poultry in district Rawalpindi.A survey was conducted for a period of 6 months in order to determine prevalence and trends of Avian Influenza H9 in the live birds markets of district Rawalpindi. A total of 355 samples were required to estimate retail shop level prevalence of avian influenza. Using systematic sampling method 14 butchers will be enrolled from 2 sentinel live bird market and they were visited weekly to collect samples for a period of 6 months. In each week 14 pooled samples from 70 birds (a pool of 5 swab samples and 2 serum samples from each shop was collected) was collected from both sentinel sites. Tracheal/oropharyngeal swabs will be collected from live and apparently healthy poultry birds then stored properly at 4°C (24-48hours) until processed. Data was collected from the shopkeeper in a face to face interview. A detail predesigned questionnaires were filled after taking written consent from the owner. The sample collected during the survey live birds markets of Rawalpindi district were processed for laboratory analysis. Real time RT-PCR and HA and HI tests for avian Influenza virus were conducted to diagnose sample for AIV. The proportion estimate with 95% C.I (confidence intervals) of the overall prevalence was computed by using R software. Pattern of influenza infection in live bird markets were estimated with reference to space and time. Descriptive analysis was conducted (i.e. mean, proportion) to answer four epidemiological W’s i.e. what, who, when and where. In present study, sero-positivity against H9 AIV was determined in district Rawalpindi. Haemagglutination (HA) assay was performed and HA titer of 1:256 was calculated, the dilutions of 8HAU was 1:32.Serum samples (n=784) were tested by HI. Out of these 784, 306 sera samples were positive (HI titer>1:8) from 2 preselected sentinel markets and 14 poultry shops, while 478 were negative (HI titer <1:8) for AI. The highest antibody titer was 1:64. The period sero-prevalence was 39.03% (95% CI: 35.41- 42.44).Results showed thatnumber of positive was high in the month ofOctober and November then slight decreased in the month of December, January and February after that it again increased in the month of March, April and May.During the study a week with one positive result was considered positive and a week was declared negative when all samples were negative. Results showed that minimum 4 and maximum 22 weeks remained positive sample results.Fourteen shops were followed up for 28 weeks and from each shop the study 56 sera samples were collected throughout the study. In 14 shops, highest prevalence was 48.21% and lowest was 28. 57% It is concluded that avian influenza is circulating in Live Bird Markets in district Rawalpindi. This process shared that this market could perpetuate and transmit avian Influenza to Human. So these markets are the hot spot of avian influenza infection. Availability: Items available for loan: UVAS Library [Call number: 2798-T] (1).

15. Physico-Chemical Analysis Of Milk From Different Milch Species (Cow, Buffalo, Camel)

by Tahira Jamil (2015-VA-595) | Dr. Sanaullah Iqbal | Haroon Jamshid Qazi | Dr. Muhammad Tayyab | Muhammad Asif Ali.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2017Dissertation note: SUMMARY Milk is described as almost a complete food as it contains all the essential nutrients in balanced quantity. Milk is a complete basis of proteins, fats and dietary energy and there are several factors that can effect the composition of milk. Factors such as (seasonal changes, feed, environmental changes, lactation, milking durations) and variations in analytical methods such as (evaluating proteins, fats, total solids, ash and moisture) can also lead to differences in results. According to FAO STAT 2010, despite the fact that Pakistan ranked among top five milk producing countries in the world, no study has been made so far that is composed of complete data based on physico-chemical analysis of milk composition of various species with respect to seasonal changes. Milk samples were collected from three different species from UVAS Pattoki Campus i.e. cow, buffalo and camel in morning and evening time. The samples were then sent to UVAS Lahore Campus. These samples were analyzed to obtain different compositional parameters of milk which includes LR, fat, protein, SNF, TS, Ash, Moisture, pH, COB and APT. In the present study, the results showed that the LR, fat, SNF, TS, Proteins, ash, moisture and pH showed no signifgicant differences when studied between the groups by independent sample t test. All results were statistically non-significant i-e p>0.05. Whereas when results of each sample were studied individually throughout the year by descriptive statistic, it was found that samples of cow, buffalo and showed high content of fats, SNF, TS and protein during the summer season and lower in winter season. Other parmeters like ash, moisture, pH also had significant change throughout the year. The monthly results were found to be statistical significant at p<0.05. COB and APT were analyzed as soon the samples arrived the laboratory. So no clotting or precipitations were observed in the sample and gave the negative results throughout the year. Thestudy was helpful in generating yearly data that was used in comparing the physico-chemical variations in morning and evening samples of milk among different milk producing species (cow, buffalo, camel) on the basis of seasonal changes. Conclusion: The directive of the current research was to analyze the physico-chemical parameters from the morning and evening samples of milk of three milk producing species (cow, buffalo, acmel). It was concluded from the results that no significant differences were found within groups of each sample. Whereas when the analysis were conducted on monthly basis throughout the year, it was determined that fat content of the samples of cow, buffalo and camel was high during the summer season. There are several reasons for this such as lactation, feed composition, milking timings, seasonal variations. SNF, TS and protein contents were directly related to fat. It was possible to state that when the fat of milk was higher the solid not fat, total solids and protein contents were also higher. However the other contents of milk such as ash, moisture, pH, COB and APT were not significantly affected by these factors. Limitations:  Diet is also an important factor that could affect the composition of milk. This factor can also be researched along with seasonal changes.  Different geographical regions affect the milk composition of animals. This is also another factor of interest.  Physiochemical changes of sheep, goat and humans can also be analyzed on the basis of seasonal changes. Milk is described as almost a complete food as it contains all the essential nutrients in balanced quantity. Milk is a complete basis of proteins, fats and dietary energy and there are several factors that can effect the composition of milk. Factors such as (seasonal changes, feed, environmental changes, lactation, milking durations) and variations in analytical methods such as (evaluating proteins, fats, total solids, ash and moisture) can also lead to differences in results. According to FAO STAT 2010, despite the fact that Pakistan ranked among top five milk producing countries in the world, no study has been made so far that is composed of complete data based on physico-chemical analysis of milk composition of various species with respect to seasonal changes. Milk samples were collected from three different species from UVAS Pattoki Campus i.e. cow, buffalo and camel in morning and evening time. The samples were then sent to UVAS Lahore Campus. These samples were analyzed to obtain different compositional parameters of milk which includes LR, fat, protein, SNF, TS, Ash, Moisture, pH, COB and APT. In the present study, the results showed that the LR, fat, SNF, TS, Proteins, ash, moisture and pH showed no signifgicant differences when studied between the groups by independent sample t test. All results were statistically non-significant i-e p>0.05. Whereas when results of each sample were studied individually throughout the year by descriptive statistic, it was found that samples of cow, buffalo and showed high content of fats, SNF, TS and protein during the summer season and lower in winter season. Other parmeters like ash, moisture, pH also had significant change throughout the year. The monthly results were found to be statistical significant at p<0.05. COB and APT were analyzed as soon the samples arrived the laboratory. So no clotting or precipitations were observed in the sample and gave the negative results throughout the year. Thestudy was helpful in generating yearly data that was used in comparing the physico-chemical variations in morning and evening samples of milk among different milk producing species (cow, buffalo, camel) on the basis of seasonal changes. Conclusion: The directive of the current research was to analyze the physico-chemical parameters from the morning and evening samples of milk of three milk producing species (cow, buffalo, acmel). It was concluded from the results that no significant differences were found within groups of each sample. Whereas when the analysis were conducted on monthly basis throughout the year, it was determined that fat content of the samples of cow, buffalo and camel was high during the summer season. There are several reasons for this such as lactation, feed composition, milking timings, seasonal variations. SNF, TS and protein contents were directly related to fat. It was possible to state that when the fat of milk was higher the solid not fat, total solids and protein contents were also higher. However the other contents of milk such as ash, moisture, pH, COB and APT were not significantly affected by these factors. Limitations:  Diet is also an important factor that could affect the composition of milk. This factor can also be researched along with seasonal changes.  Different geographical regions affect the milk composition of animals. This is also another factor of interest.  Physiochemical changes of sheep, goat and humans can also be analyzed on the basis of seasonal changes. Availability: Items available for loan: UVAS Library [Call number: 2884-T] (1).

16. Relationship Among Physical Reproductive Charecters And Semen Quality Parameters At Different Age Groups Of Sahiwal Bulls

by Muhammad Asif (2015-VA-1077) | Dr. Aijaz Ali Channa | Prof. Dr. Nasim Ahmad | Dr. Muhammad Avais.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: The present study demonstrate the relationship among physical reproductive characters and semen quality parameters at different age groups of Sahiwal bulls. The standard breeding soundness evaluation criteria is not set for the screening of Sahiwal bulls. Artificial insemination is a tool for rapid genetic improvement. Sahiwal bulls used as AI sire should be screened through standard breeding soundness evaluation criteria. Therefore the present study has been designed to determine objectives like: (1) to determine the relationship among physical reproductive characters and semen quality parameters at different age groups of Sahiwal bulls (2) to compare the semen quality parameters of pendulous with non-pendulous Sahiwal bulls. The study was carried out during July-October 2016 at Semen Production Unit Qadirabad district Sahiwal. The Sahiwal bulls (n=91) maintained at SPU as a regular semen donor bulls, having no reproductive disorder were selected for this study. These bulls were divided into four different age groups G1 (24-48 months, n=8), G2 (49-72 months, n=50), G3 (73-96 months, n=27) and G4 (97-108, n=6). The evaluation of the bulls were done for BW, SC and DPS. Semen quality parameters (ejaculate volume, sperm concentration, motility, percent live sperm and morphological abnormalities) were evaluated and compare between different groups. Mean BW and SC increases (P˂ 0.05) with the age of bulls, but DPS is not correlated with the age means it does not increases with the age. Ejaculate volume (3.58±0.1, 3.92±.05, 3.97±.06, 4.24±0.15 ml), sperm concentaration (870.6±14.1, 897.4±6.5, 899.5± 8.0, 856.9±20.2 x 10x6 /ml) increases with age. However motility remain more or less unchange with age . The percentage of live sperms were 75.06±0.3, 75.75±0.1, 75.96±0.1, 77.09±0.4 respectively and percentage of morphological abnormalities were Summary 32 19.57±0.6,17.61±0.2,15.80±0.2,16.69±0.4 repectively. The morphological abnormalities were higher in younger bulls as compered to older bulls. It can be concluded that the BW, SC, ejaculate volume, sperm concentration (except in G4 due to small sample size) increases as that the age increases, whereas the other semen quality parameters like individual motility, motility after dilution and individual motility after freezing looks to be more or less independent of age. The DPS is not correlated with the age, it may be a genetic character. In comparison of semen quality between pendulous and non-pendulous bulls, the ejaculate volume and sperm concentration were significantly higher in pendulous bulls instead of non-pendulous bulls (in G2 and G3) and nonsignificantly higher (in G1 and G4). While comparing the other semen parameter like motility percentage (individual, after dilution and after freezing) between pendulous and non-pendulous bulls the results remain non-significant. The findings of this study direct us to focus more on nonpendulous bulls instead of pendulous bulls, the reason behind that is the pendulous bull’s needs extra managemental care as they are more prone to injuries and perpetual infections. These findings could be used to set the selection criteria for Sahiwal bulls. Availability: Items available for loan: UVAS Library [Call number: 2922-T] (1).



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