Compaiativ Efficacy Of Different Electrolyte Solutions On Heat Stress And Their Efiect On Hematology And Blood
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; Literary form:
Publisher: 2011 Dissertation note: The present project had been designed to study the effect of heat stress on cattle calves and to evaluate the efficacy of electrolytes solution (Normal Saline and Ringer Lactate) on various blood parameters. Five groups of calves comprising 10 in each group were selected for experimental study. Group A: Affected calves with heat stress were provided shade after taking TPR and the effect of shade were checked after one hour. Group B: Heat stressed calves of same age group were given Normal Saline IV according to their body weight and the effect were checked through TPR, hematology and blood electrolyte. Group C: Heat stressed calves of same age group were given Ringer's Lactate IV according to their body weight and the effect were checked through TPR, hematology and blood electrolyte. Group D: calves of same age group affected with heat stress were taken as the positive control. Group E: calves of same age group were normal healthy calves (negative control). Temperature was taken at regular intervals of one hour daily. Respiration was observed by placing the hand in front of nostrils. Heart rate was observed by stethoscope daily in morning and evening. The blood sample of each calf was collected both for control and experimental animals through disposable syringe from jugular vein. The blood was shifted to University Diagnostic laboratory, University of Veterinary and Animal Sciences Lahore. The samples were taken before and after therapeutic trials. Blood samples were taken for blood electrolyte examination and hematology. Serum of the blood was separated by centrifugation for electrolytes measurements. The flame photometer was utilized to measure the serum sodium (Na+) potassium (K+) Chloride (Cl+) and Bicarbonate (HCO3) concentration. The physical sign of experimental group before cooling were noted .sever sweating and panting were observed under physical sign. The pulse rate, respiration and rectal temperature of experimental group before cooling were increased. Changes found in CBC and blood electrolytes like sodium, potassium, chloride and bicarbonate were measure by flam photometry.
These all observation showed that the animal of experimental group before cooling were suffering from electrolyte imbalance ,but it was not so serious which may result in death of the animal, however the persistence of that condition might result in heat stroke which is often lethal.
It is concluded that serum electrolyte concentration, CBC and pulse rate, respiration and rectal temperature help in accessing the condition of animal suffering form the heat stress. From the present study it can be concluded that heat stress cause changes in biochemical and Hematological parameters in calves. These changes can be overcome by giving animal's fluid therapy and by providing good shade in hot summer. Further studied are required to Conducted on other species of animals to understand the effect of heat stress .Other biochemical and hematological parameters should be studied in bovine calves and other animals for the better understanding the effects of heat stress.
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Molecular Diagnosis Of Bovine Anaplasmosis In District Lahore
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Publisher: 2011 Dissertation note: The present study was designed to determine diagnosis and infection percentage of Bovine anaplasmosis in cattle and buffalo of different age groups in and around District Lahore, and to study the comparative efficacy of diagnostic methods that is Polymerase chain reaction (PCR) and Microscopic Examination. For this purpose 160 blood samples were collected from cattle and buffaloes ,randomly from eight villages , during the month of May, June, July, August of 2010 in and around District Lahore.80 samples were collected from cattle and 80 were collected from buffaloes and these samples were further categorized into two age groups that is 40 samples were collected from calves of 1 month to 6 month of age and 40 samples were collected from calves of 7 month to 12 month of age of each species. Screening was done by blood smears, stained by Giemsa'wright staining technique and later the blood samples from the same animals were also processed by PCR.
The blood smears showed Anaplasma marginale as dense , round, deeply stained body, approximately 0.3-1.0um in diameter. Most of them were located on or near the margin of the erythrocyte.On the basis of Microscopic examination overall 11.25% (18\160) prevalence was recorded. On the basis of polymerase chain reaction (PCR) prevalence of Anaplasma marginale 25.6%(41\160) was recorded, showing the presence of carrier animals in District Lahore. The blood smears showed maximum prevelance in cattle of age 7 months to 12 months of age that is 20% (8\40) than animals of age 1 month to 6 month of age 10% (4\40).The blood smears showed maximum prevelance in buffalo calves of age 7 months to 12 months of age that is 10% (4\40) than animals of age 1 month to 6 month of age 5% (2\40). The blood smears showed that the prevelance of Bovine anaplasmosis is more in cattle 15% (12\80) than buffalo 7.5% (6\80). The overall prevalence 25.6% (41\160) was recorded for Bovine anaplasmosis , during summer season on the basis of PCR. The Polymerase chain reaction showed maximum prevelance in cattle of age 7 months to 12 months of age that is45 % (18\40) than animals of age 1 month to 6 month of age 20% (8\40). The Polymerase chain reaction showed maximum prevelance in buffalo animals of age 7 months to 12 months of age that is 27.5% (11\40) than animals of age 1 month to 6 month of age 10% (4\40). The Polymerase chain reaction showed that the prevelance of Bovine anaplasmosis is more in cattle 32.5% (26\80) than buffalo18.75 % (15\80).The results have shown high efficacy of PCR as compare to Microscopic Examination.
It is anticipated that present study was proved helpful in diagnosis of Anaplasma in infected as well as in carrier animals in District Lahore , and will be beneficial for further study.
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Evaluation Of Risk Factors And Molecular Diagnosis Of Dermatophytosis In Dogs
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Publisher: 2016 Dissertation note: Dogs are most kept and beloved pets in Pakistani society. Dermatophytosis is among the common disease of the pets. Many predisposing factors are involved in development of clinical cases of dermatophytosis including climatic conditions, housing condition of dogs and physical attributes such as coat hair size. Dermatophytosis is not only of concern as being infection of pets but also of its zoonotic importance hence it is very crucial to diagnose dermatophytic infection well in time. Dermatophytosis is caused by Dermatophytes,Microsporum, Trichophyton and Epidermophyton, the fungal species. It is difficult to diagnose the Dermatophytosis from other skin infections by routine tests in most of the cases especially subclinical. Polymerase Chain Reaction (PCR) is advanced and the most reliable technique to detect genome of Dermatophytes even in minute quantities specifically and can efficiently detect the presence of any Dermatophyte specie on the skin of dog. The current study was planned to develop and validate a diagnostic assay which could be able to detect and distinguish tree important dermatophytes species including Microsporum, Trichophyton and Epidermophytonby a uniplex PCR reaction. Analysis of involvement of certain predisposing factors in dermatophytosis was second goal to be worked on in this study. Samples of suspected pet dogs (n=50) were collected by scraping the skin at affected areas over skin. DNA was extracted from the skin scraping samples by organic Phenol Chloroform Isoamyle Alcohol method. Primers, specific to the 18-S ribosomal RNA region of genomes of the Dermatophytes, were designed after alignment of available sequences of Microsporum,Trichophyton and Epidermophyton at NCBI. Annealing temperature and recipe of PCR reaction was optimized by gradient PCR in BIO-Rad thermal cycler. Amplification reaction of all samples collected was carried out as per optimized reaction conditions, afterwards. Amplified products obtained were subjected to genotyping by agarose gel electrophoresis for size based separation of the amplified products. The specific amplified bands of desired genomic region of dermatophytes were seen in UV light transilluminator. The data of results of predisposing factors involved in dermatophytosis wasanalysedby using Pearson’s chi squared test with the help of Statistical Package for the Social Science (SPSS) Program.
Genome specific product sizes of Microsporum and Trichophyton i.e. 366 bp and 351 bp in respective positive samples were observed. Out of 50 suspected samples 46 samples were positive for dermatophytosis out of which 38 samples (82.6%) were positive for Microsporum, 6 samples (13%) for Trichophyton and 2 samples (4.4%) were positive for both Microsporumand Trichophyton.
This study will help to validate a diagnostic technique for Dermatophytosis with greater efficacy and reliability. Moreover, this investigation may become basis for the future research activities in this field in Pakistan.
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Comparative Pathological Association Of Slc11a1 (Nramp1) Gene With Susceptibility And Resistance To Brucellosis In Local Pakistani Buffaloes
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Publisher: 2017 Dissertation note: Nearly 32.7 million population of buffaloes present in Pakistan which makes about 15% of the world buffalo population. For water buffaloes Brucellosis is a pricey disease (Bubalus bubalis). In Pakistan 5 known breeds of buffalos are; Ravi, Nili, Kundi Nili-Ravi and Aza Kheli. The study was conducted on 200 animals of two breeds including Nilli-Ravi and Kundi around farms of Punjab. 200 serum samples from two breeds were screened for brucellosis. Dormant infections and lengthy incubation of the pathogens bounded the effectiveness of programs based upon the elimination of infected animals. As comparative genomics is playing a pivotal role in the genetic dissection of complex traits such as infectious diseases resistance using this information we can oppressed for disease resistance with genetic choice as a method to the regulator of brucellosis in our local breeds such as Nili-Ravi buffalo. Blood sample (5mL) was collected in EDTA vacutainers from each animal. Serum from blood samples were collected in 3 mL eppendorf tubes with the help of sterile pipettes. Serum samples were screened out for brucellosis by Rose Bengal Plate Tetst (RBPT). Rose Bengal Antigen was purchased from Veterinary Research Institute (VRI), Lahore. The antigen was stored at 40C in refrigeration during the study according to manufactures recommendation. DNA quantification was performed on “Thermo Scientific NanoDrop spectrophotometer ND-2000” and by running extracted DNA on 0.8% agarose gel. Nanodrop spectrophotometer provides direct and easy measurements within a 5 second only by using just a pipette and wipe. PCR for amplification was done with a total volume of 20μl by adding primer pair and extracted DNA to the PCR master mix in following concentrations. 2μl of forward and reverse primer was taken respectively. 4 μl of PCR grade water was added and DNA was taken in 2 μl
Quantity. The total volume of master mix obtained was 10 μl. The thermocycler (Bio-Rad, CFX ™, Singapore) was set with the conditions for the total of 35 cycles of PCR. Initial denaturation of DNA samples were performed at 940 C for 5minutes. Then in 2nd cycle denaturation was performed at 94 0C for 30 seconds. Annealing step was performed at 63 0C for 30 second. Then, initial extension of DNA samples was conducted at 720 C for 30 second. Finally, extension was performed at 720 C for 5 minutes to get results. The PCR product was confirmed by running the PCR product on 1.5 % Agarose gel electrophoresis was performed at 110 Volts for 30 minutes. In Nilli-Ravi, among 12 positive samples by PCR, 08 were found resistance to brucellosis through sequencing study of Nramp 1 BB, only one sample was found to be susceptible to brucellosis through genotypic study of Nramp1 AA and 3 samples were found to be neutral may be susceptible or resistance to brucellosis. In Kundi, among 38 positive samples by PCR, 03 were found resistance to brucellosis through sequencing study of Nramp 1 BB, and 6 sample were found to be susceptible to brucellosis through genotypic study of Nramp1 AA and 29 samples were found to be neutral may be susceptible or resistance to brucellosis. This research clearly demonstrates that Kundi is more susceptible to brucellosis then Nilli-Ravi. Conclusion: We concluded that Nilli-Ravi is more resistant to brucellosis as compare to Kundi breed Natural resistance against brucellosis in cattle is linked to the Nramp 1 gene. Polymorphisms at the bovine Nramp 1 3/ untranslated region (3 UTR) are associated with natural resistance against brucellosis.
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