1.
Studies On Antigenic Homogeneity Of Fowl Adenoviruses Causing Hydropericardium Syndrome In Broilers
by Muhammad Tariq Iqbal | Dr. Mansur-ud-Din Ahmad | Dr. Irshad Hussain | Dr. M. Sarwar | Faculty of Veterinary Sciences.
Material type: Book; Format:
print
Publisher: 2006Dissertation note: Babesiosis is a highly important disease in the world, caused by the intraerythrocytic protozoan parasites of the genus Babesia. A wide range of domestic and wild animals and occasionally man are affected by this disease, which is transmitted by ticks and has a worldwide epidemiological distribution. While the major economic impact of babesiosis is on the cattle industry, infections also occurs in other domestic animals , including horses, sheep, goats, pigs and dogs.
The present study targeted the carrier cattle infected with Babesia bigemina and Babesia bovis, as they are difficult to detect because of the low numbers of parasites that occur in peripheral blood. However, diagnosis of low-level infections with the parasite is important for studying the transmission and standardizing epidemiological studies. Using the Polymerase Chain Reaction (PCR) to amplify a portion of the gene from the parasite, and tested the ability of this method to detect carrier cattle.
A study was conducted to detect the. Babesia in blood samples through PCR based techniques. A PCR assay was described which could differentiate Babesia bigemina and Babesia bovis by using specific primer in carrier cattle. Blood samples of 100 cattle were randomly analyzed with PCR assay 29 (29.0%) out of 100 blood samples were positive for babesiosis in which 18% were positive for Babesia bigemina and 11% were positive for Babesia bovis, While the Light Microscopy detected only 18 (18%) out of the same samples. The samples found positive by LM were reconfirmed during the PCR assay but no sample was found to be having both Babesia bigemina and Babesia bovis infections simultaneously.
Thus it is concluded that PCR is a reliable molecular diagnostic technique to detect low level of infections in carrier animals in a population and thus could be used as an effective screening tool for the control and eradication of disease.
Availability: Items available for loan: UVAS Library [Call number: 0928,T] (1).
2.
Molecular Detection Of Programmed Cell Death (Apoptosis In Fresh And Cryopreserved Buffalo Sper Matozoa)
by Daulat Raheem Khan | Prof. Dr. Nasim Ahmad | Dr. Irshad | Dr. Muhammad Amir Saeed | Faculty of Veterinary Sciences.
Material type: Book; Format:
print
Publisher: 2006Dissertation note: The objectives of present study were a) to validate annexin V/Pl assay, for buffalo sperm, using graded doses of camptothecin through fluorescent microscopy (Exp. 1) and b) to determine the effect of stages of cryopreservation on apoptosis (PS externalization, using annexin V/PT assay), motility and plasma membrane integrity in buffalo sperm (Exp.2). In the first experiment graded doses of camptothecin were used (n=2) in different aliquots of sperm suspension (1x106/mL) for induction of apoptosis. Higher dose levels of camptothecin (10.0 iM and 20 jiM) resulted in inducing apoptosis (P<O.05) compared to the lower (5tM) dose or control. In the second experiment semen samples (n=9, from three bulls) were cryopreserved using vapor freezing. Annexin V/PI assay for apoptosis, visual assessment for percentage motility and hypo-osmotic swelling test for plasma membrane integrity were employed at various stages of cryopreservation (i.e. fresh, after equilibration and post thaw). The mean percentage of apoptotic, necrotic and viable sperm did not differ between fresh and after equilibration stages. However, freezing and thawing increased (P<0.05) the percentage of apoptotic sperm (25.4±0.6 vs. 36.5±1 .9) while decreased (P<0.05) the necrotic (29.7±0.7 vs. 35.1±1.2) and viable sperm (37.2±1.3 vs. 32.8±1.9, (P<0.07). Similarly, freezing and thawing decreased (P<0.05) the mean percentage motility and plasma membrane integrity, compared to other stages. Based upon the difference in initial and post thaw values of two variables (percent motility and percent apoptosis) it is concluded that apoptosis contributes more than 50% sperm motility loss during the process of freezing and thawing in buffalo.
Availability: Items available for loan: UVAS Library [Call number: 0933,T] (1).
3.
Comparative Efficacy Of Different Prophylactic & Curative Measures Against Caecal Coccidiosis In Poultry
by Muhammad Imran Rashid | Dr. H. A. Hashmi | Dr. Irshad | Mr. Wasim Shehzad | Faculty of Veterinary Sciences.
Material type: Book; Format:
print
Publisher: 2006Dissertation note: To find out the therapeutic and prophylactic efficacy of toltrazuril against coccidiosis, an experiment was conducted in the Department of Parasitology, University of Veterinary & Animal Sciences, Lahore.
One hundred and sixty (160) day old chicks were divided into 8 groups, i-e, A, B, C, D, E, F, G and H each consisting of 20 chicks. Group A acted as non-infected and non-medicated control, B was infected with Eimeria tenella sporulated oocysts on day 16 and 26 @ 20000 per chick and acted as infected control. Similarly, all other members of different groups were also infected at the same rate. Members of Group C were administered toltrazuril orally in drinking water @ 7 mg/ kg body weight on day 18 & 19 and 25 & 26. Members of Group D were given toltrazuril @ 3.5 mg/kg body weight, of Group E @ 1.75 mg/kg as in group C & D, of Group F toltrazuril was given as prophylactic @ 7mg/kg on day 3 & 10. Members of Groups G & H were given irradiated (IEV) and formalized (FEV-48) Eimeria tenella vaccines respectively. IEV was orally given on day 3 & 10 whereas FEV was only given on day 10 of age.
The result indicated that maximum reduction of OPG counts (94.28 %) occurred in members of group D which was given half the dose (3.5 mg/kg) of toltrazuril, Group E (Administered quarter dose) was placed at no.2 and the reduction in this Group occurred as 93.71 %. The group G (IEV immunized) was placed at no. 3 by reducing 89.71 % mean OPG counts as compared to control Group B.
In terms of reduction of OPG counts other Groups were placed at no.4 (Group F given full dose prophylactic), no.5 (Group H immunized with FEV-48) and Group C showed the poorest performance (63.42 %) which was given full dose was placed at no. 6.
In terms of Mean body weight gains as compared to control (healthy) Group A, Group D was placed at no.1, Group E at no.2, Group G at no.3, Group C at no. 4 Group F at no.5, Group H at no.6 and Group B which acted as infected and non-medicated was placed at no.7. Ultimately, it was concluded that toltrazuril @ 3.50 mg/kg or immunization with IEV (irradiated vaccine) would be the best solution to the coccidiosis problem
Availability: Items available for loan: UVAS Library [Call number: 0941,T] (1).
4.
In Process Quality Control Factors Affecting Efficacy Of Hydropericardium Syndrome Virus Vaccine
by Muhammad Danish Mehmood | Prof. Dr. Khushi Muhammad | Dr. Irshad Hussain | Prof. Dr. Zafar | Faculty of Veterinary Sciences.
Material type: Book; Format:
print
; Literary form:
not fiction
Publisher: 2007Dissertation note: The objective of this project was to study in process quality' control factors affecting the efficacy of Hydropericardium Syndrome virus vaccine in broilers. The parameters studied were mortality and protection percentage, seroconversion and affect of HPS infected liver homogenate vaccine on body weight gain of broiler birds. In this different vaccines were prepared from HPS infected liver homogenate having different biological titer (105.6,104.6 and 103.6 units of infectivity Animal Lethal Dose 50-ALD50) inactivated with 0.15% formalin. The other type of Hydropericardium Syndrome vaccine was prepared from chicken embryo hepatocytes having biological titre 1036 tissue culture infective dose-TCID50. At day 14th of age, groups Al, A2 and A3 were vaccinated with HPS infected liver homogenate aqueous based vaccines having different biological titre. While groups Bl, B2, B3, B4 and B5 were vaccinated against 20, 25, 30, 35 and 40 doses per gram of HPS infected liver homogenate vaccine and groups Cl, C2, C3 were vaccinated with lanolin based HPS vaccine, gel based HPS vaccine, montanide based HPS vaccine respectively. Similarly group Dl, D2 were vaccinated with HPS virus chicken embryo hepatocyte vaccine and HPS liver homogenate vaccine respectively. The group El, E2 and E3 were vaccinated with HPS virus infected liver homogenate vaccine containing preservative (thiomersal sodium) stored for 30, 60 and 90 days respectively. The birds in group F served as uninnoculated controls. The HPS infected liver stored for 0-45 days at -20 C and processed for determination of its biological activity at fortnightly interval. It was observed that HPS vaccine containing more than 104.6 and 105.6 units of the immunogen provided protection to 100% in vaccinated birds. The 20 doses and 25 doses of the gel based HPS vaccine per gram of the liver developed 90% protection in vaccinated birds. Montanide based HPS vaccine provided 100% protection while HPS virus infected homogenate vaccine containing thiomersal sodium provided 80% protection up to 90 days and HPS virus chicken embryo hepatocyte vaccine provided 40%protection in the vaccinated birds. Se3rum samples were collected form all groups on 14 and 28 day post vaccination and subjected to AGPT for seroconversion. Each serum sample when monitored for anti-HPSV antibodies through agar gel precipitation test, showed undetectable titre.
Availability: Items available for loan: UVAS Library [Call number: 0960,T] (1).
5.
In Vitro Production Of Aflatozin (B1), Its Purification, Estimation And Biodegradation
by Muhammad Hanif Khan | Dr. Irshad Hussain | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: Book; Format:
print
Publisher: 2007Dissertation note: In this study a toxigenic strain of A. parasiticus was used for the production of the aflatoxin Eli. This strain of the fungus was taken from naturally growing fungal samples such as bread, rice, vegetables, fruits, maize etc. from various locations in and around the university. The unpurified sample was culture-purified on Sabouraud Agar through repeated culture technique. This strain was maintained on Sabouraud agar slants at 4 °C. The purified sample of this fungus was harvested with 0.01 % Tween 80 then inoculated to the autoclaved broken rice. The rice was incubated at 28 °C in a dark room for a period of' 7 days and shaken two times a day. After 7 days of incubation the rice as autociaved at 121 °C in order to inactivate the fungus. The rice was dried and crushed to powder in a blender. The toxins were extracted through chloroform methpd and evaporated to dryness in water bath at 6 °C. The toxins were again dissolved in I ml ehiorokrm and stored at 4 °C for further use in the study. The toxins were then purified on silica gel plate in order to get the AFBI only through comparison with the AHII standard. 'l'he aflatoxin BI band was scratched from the silica gel plate and subjected to centrifugation in order to remove the silica gel. The purified Bi toxin was then dissolved in chloroform for further use. The AFB1 was then identified and qilani i lied through the fol lowing three methods namely,
1)High perlirmance Liquid chromatography
2) Thin layer chromatography
3) Spectrophotornetry
The sensitivity of the above three methods was determined and compared which was 1 .3 ng/ml for HPLC. 6.3 ng/ml forTLC and 40 ng/mI for Spectrophotometry. The purified AFBI was then estimated by HPLC the amount of which was 238 ± 9.8 jig / ml. however the toxins estimated through TLC were 127.8 ± 24 jig/mi and spectrophotometry were 26 ± 4jig/ml ml. The HPLC method was proved to be the most sensitive method for detection and estimation of the aflatoxin as compared to the other two methods. The HPLC, estimated toxins were then checked for their biodegradation for this purpose the AFBI were kept at three different temperatures and dissolved in three different solvents. As the toxins had previously been dissolved in the chloroform so 400 jil of the chloroform was evaporated to dryness in order to dissolve them in their respective solvents. The toxins were dissolved in 400 j.tl of chloroform, acetone and methanol and kept at 25, 4 and -20 °C. These toxins were then stored for a period of four weeks and the tested for their biodegradation after four weeks by HPLC.
The temperature at which the toxins showed less degradation was determined. That temperature at which the toxins showed less degradation was - 20 °C and the amount of the toxin at this temperature after four weeks of storage was 197 jig/ml. The solvent in which the toxins exhibited less degradation was acetone at all the three temperature conditions (25, 4 and -20 o() I lowever chloroform at 25 °C was exhibiting less degradation as compared to the other two solvents. Methanol was proved to be the least good solvent because the toxin was showing degradation in this solvent. The acetone at -20 CC was the most appropriate solvent for toxins storage.
In second experiment the fresh toxin was dissolved in acetone and tested fbr their degradation on weekly basis by HPLC. The toxin level at the beginning of the experiment was 447 ± 9 ug/mI which when detected after a weak was 398.67 ± 9.8 jig/mI. The same toxin when tested after 2 weak period storage exhibited a level of 295.9 ± 20 ug/mi followed by 265.7 ± 3Ojig/ml. after the end of the third weak indicating that there was an appreciable level of toxin degradation. The toxin tested on the fourth week of storage were 267± 31 ig/ml which exhibited no degradation as compared to the degradation of the third week.
Availability: Items available for loan: UVAS Library [Call number: 0977,T] (1).
6.
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: Book; 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).
7.
Identification And Genotyping Of Vp1 Genses Of Fmd Viruses
by Atia Bukhari | Prof. Dr. Irshad Hussain | Prof. Dr. Khushi Muhammad.
Material type: Book; Format:
print
Publisher: 2009Dissertation note: Within two decades after its first report in 1954 from Pakistan, Foot and mouth disease has become endemic in the country and poses a serious threat to large as well as small ruminant population. Foot and Mouth Disease (FMD) is prevailing in cattle and buffaloes and is caused by either 0, A, Asia-i serotype of the FMD virus in Pakistan. The present study was undertaken to study the mutation rate of FMD virus and also molecular typing of the strains prevalent in Pakistan was done.
A total of 60 samples from buffalo and cattle were collected from five districts of Punjab including Lahore, Faisalabad, Sialkot, Okara and Sheikhupura. Soon after extraction of their RNA, all of them were reverse transcribed and then subjected to amplification by using different sets of the primers including universal as well as serotype specific primers. Then their VPI portions were amplified by using VP1 specific primers. Among 60 samples, 48 were positive with universal primers. Other 12 samples were not amplified with these primers hence not processed.
Among 48 FMD positive samples, 24 were positive with serotype 0 specific primers, 16 with serotype Asia-i and remaining 8 were positive with serotype A specific primers. After their amplification, the amplicons were run on the gel. These amplicons were extracted by using DNA extraction kit. After their purification, they were sent to Macrogen® (Seopl, Korea) and Centre of Excellence for Molecplar Biology, Pakistan (CEMB) for sequencing. Each amplicon was sequenced thrice and the consensus sequence was established eliminating sequencing errors.
Sequence identity and multiple sequence alignment of molecular sequences (nucleotide and amino acids) were performed with Clustal W algorithm (Thompson et al., 1994). Neighbour joining trees were constructed by using MEGA version 4.0 (Kumar et al., 2004). Nucleotide distance matrices were computed by Kimura two parameter algorithm based on the total nucleotide substitutions and evolutionary trees for VP1 genes were constructed.
For FMDV serotype '0' phylogenetic analysis, 14 VPI sequences from various field isolates were compared with some previously published Pakistani FMD 0 type VP1 specific sequences available with GeneBank and some recently published VP1 sequences reported by countries bordering with Pakistan including India, Iran and Afghanistan Similarly, 12 VP 1 sequences of FMDV serotype Asia-I isolates of this study were compared with previously published sequences and their phylogenetic relationship was established. However, the sequencing results of serotype A were inconclusive and were not included for phylogenetic analysis. Three sequences of three locally available FMD vaccines were also studied and compared with the outbreak strains.
Polymerase chain reaction was optimized with respect to MgCI2, buffer pH, annealing temperature, primer concentration, template concentration, and Taq polymerase. A concentration of 2.5 mM of MgCl2 resulted in the best amplification of the target sequences (Figure 1). The buffer with pH 8.8 yielded the best results (Figure 2) Although, the suggested annealing temperatures for various primers (of various serotypes) ranged from 48 °C to 63 °C, however, a temperature of 56 °C was found to be the best with all sets of primers (Figure 3). The best intensity DNA bands were observed with 0.3 pM concentration of the primers (Figure 4). Moreover, the best cDNA template concentration giving optimum amplification was found to be 3.0 p1 per reaction (Figure 5). Lastly, a concentration of 0.5 U of Taq polymerase was not sufficient for amplification of cDNAs, however, 1.0 U of enzyme was found to yield better amplification (Figure 6).
VP 1 DNA sequences of six previously published Pakistani FMD serotype 0 strains were analyzed phylogenetically with VP 1 DNA sequences of 14 isolates of the study. Serotype 0 isolates of this study distributed themselves into two distinct clusters (Figure 19). First cluster comprised of Sheikhupura 1 and 2, Muridkey 1, Raiwind 1, Nankana 1, Gujranwala 1 and Gujrat I isolates (Figures 19 and 20), whereas the second cluster included Depalpur 1, Sahiwal 1, Okara I, Multan 1, Toba 1, Faisalabad I and Pattoki 1 isolates (Figures 19 and 21). The first cluster was found to be associated with previously published Pakistani isolates of 2006 mostly. However, it also showed association with Afghanistan's isolates of 2004 (Figure 20). The second cluster seemed to be mostly related to previously published Pakistani isolates of 2003 (Figure 21). The overall grouping of the 14 sequences, when compared with each other, depicted a three clustered phylogram (Figure 22). Serotype 0 isolates from Depalpur, Sahiwal, Okara, Multan, Pattoki, Toba Tek Singh and Faisalabad grouped together into a clan and had more than 85% sequence similarity with each other. The second cluster consisted of isolates of Sheikhupura, Nankana, Raiwind and Muridkey. These sequences had more than 86% similarity with each other. The third cluster consisted of only two isolates which were 100 % similar to each other. However the third cluster had only 74 % sequence similarity to cluster I and 73 % sequence similarity when compared with cluster 2.
When the phylogenetic relationships with previously reported isolates of Asia 1 was evaluated, FMD Asia I isolates of this study were found to be scattered into two distinct groups (Figure 16). Group one consisted of isolates of Lodhran, Toba and Hafizabad that were more closely related to Indian isolates sharing more than 98% identity with each other and more than 94 % sequence identity with isolates of Indian 2001 to 2004 (Table 5 and Figures 16 and 17). However, they shared more than 86% sequence similarity with Pakistani isolates of 2002-2005 (Table 5). Group two comprised of isolates of kasur, Lahore, Pakpattan, Okara, Faisalabad, Jhang, Rahim Yar Khan, Bahawalpur and multan alongwith vaccine A and B (Figure 16). The isolates of group 2 were found to be closely associated with previously published isolates of Pakistani and Afghani origin of year 2003 and 2004 (Figures 16 and 18). Collectively, they shared an overall 70% sequence identity with each other. However, isolates of Bahawalpur, Rahim Yar Khan and Multan shared more than 98% similarity with each other, a measurement of close relationship denoting a likely common origin as one clan or dade. Similarly, isolates of Pakpatan, Faisalabad, Okara, Kasur, and Lahore shared 88% sequence identity with each other and qualified as one clade.
Although, overall amino acid sequence similarity of our isolates was not strikingly different from that of the published isolates, however, amino acid substitutions with dissimilar properties were found with a scattered pattern of distribution. For example, 15th amino acid residue which is hydrophilic in the previously published isolates had a substitution with a hydrophobic amino acid residue in our three isolates namely Sheikhupura 2, Muridkey I and Raiwind I (Figure 25). Similarly, 14th amino acid residue which is hydrophobic in nature was found to be replaced with a hydrophilic one in our last five isolates. Amino acid residue number 13 (Figure 25) had a substitution with a hydrophobic residue in some of our isolates etc. etc. It is interesting to note that such substitutions with amino acids having dissimilar properties have also been found, albeit at lower rate, in previously published sequences by many researchers (Figure 25).
A comparison of the deduced amino acid sequences in the critical VP I region of FMD serotype Asia I revealed that most of this study isolates shared very high homology with sequences of Vaccine A. However, the sequences of isolates of Lodhran, Hafizabad and Toba did not match much with that of either vaccines, A or B (Figure 23). Sequences of Vaccine A had a "K" which seemed to be replaced by a "T" in the sequences of most of the isolates. Considering the properties of various amino acids, this change does not signify a major shift in the three dimensional picture of the protein as K is a lysine, a positively charged amino acid, whereas a T is threonine, a hydrophilic amino acid in nature. Next substitution in most of the isolates is a "P" for "A" in comparison to the vaccines. Again, it is not a significant change as both P and A share the same property, hydorphobicity. Similarly a K with an R can be substituted without much change in the overall shape of the protein molecule. Next amino acid substitution is a leucine instead of methionine. Again both are hydrophobic in nature; hence their impact on the overall picture is minute, if at all. However, glycine and arginine are two very different amino acids; the former is a hydrophobic amino acid whereas the latter is positively charged one. Such amino acid substitutions may have the potential to make a major impact in terms of the epitopic differences in the capsids of vaccinal and field viruses. A comparison of the deduced amino acids of FMD serotype 0 isolates also exhibited such changes with the vaccinal virus (Figure 24).
Of the three hyper immune sera raised against three different vaccines in rabbits, only one vaccine induced a measureable immune response yielding good precipitation line against various FMD virus antigens.
In summary, RT-PCR for diagnosis of serotypes A, 0 and Asia 1 of FMDV was optimized and could be used for prompt and precise diagnosis of FMD in the country. Although, RT-PCR data pertains to bovines in the current project, but PCR optimization parameters are equally applicable to FMDV infections in other FMD susceptible animal species such as sheep and goat. The combination of PCR and sequencing of the VP1 gene to detect and analyze FMDV in disease outbreaks is fast (less than 6 hours for PCR and about 24 hours for sequencing), and it can give an accurate immunologic characterization of the virus, thus providing a rational basis for choice of vaccine. In fact, the molecular epidemiology of field isolates is a powerful tool to monitor the circulation of viruses (Saiz et al., 1993).
Secondly, various isolates of serotypes 0 and Asia 1 were sequenced along with some vaccinal strains. Sequence similarity tree analysis indicated that most of our isolates were closely related to previously reported Pakistani isolates and to those of neighboring countries such as India, Afghanistan and Iran. Additionally, amino acid sequence similarity data of major immunogenic site that forms 13G-13H loop in FMDV serotypes revealed that serotype Asia 1 vaccinal strain and Asia 1 isolates of this study possessed high degree of similarity suggesting a likely host immune response against the vaccine that may afford some protection against most field isolates of serotype Asia 1 type. Lastly, of three vaccines tested, only one was found to afford protection against field isolates of FMDV suggesting more work on vaccine issue in the country.
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8.
Effect Of Lactobacillus Bulgaricus On Cutaneous Wound Healing In Mice
by Sonia Akram (2009-VA-160) | Prof. Dr. Masood Rabbani | Prof. Dr. Irshad Hussain | Dr. Hafsa Zaneb.
Material type: Book; Literary form:
not fiction
Publisher: 2016Dissertation note: Integrity loss of major part of the skin as a result of injury, surgery, or sickness can lead to a major disability or even death. The major complication which comes in healing process is infection. Besides reports of side effects, a diversity of chemical agents is in use to facilitate wound healing process. Use of probiotics for the purpose is an approach of altering the microbial environment of wound where beneficial bacteria compete with pathogenic bacteria. Lactobacillus bulgaricus extracellular polymeric substance (EPS) contains high levels of phosphate groups which help activate macrophages and lymphocytes.
Lactobacillus bulgaricus was isolated from sources like: homemade curd, yogurt, and commercial probiotic product (Protexin, UK) as per WHO guideline. For bacterial identification, the isolates were further characterized using biochemical tests. Male mice (n= 60) were kept and divided into four groups having 15 mice in each group (A to D). Experimental treatments using suspension of Lactobacillus bulgaricus, pure honey and sterile normal saline were applied to surgically created wounds on skin of mice belonging to three groups (A, B, and C), whereas, one group (D) was left untreated. After application of treatments, tissue samples were collected from these groups, processed and examined for histopathology. Increase in number of macrophages, neutrophils and fibroblasts at wound site of mice of all groups, was recorded during histopathological examination of H & E stained slides. Results were tabulated using one-way ANOVA.
Experimental groups A and B (L. bulgaricus and honey) showed significant increase in the cells as compared to control and negative control groups (C and D). L. bulgaricus treated group
CHAPTER 6
SUMMARY
Summary
61
showed marked healing of cutaneous wound than control and negative control groups. It was observed that in L. bulgaricus treated group there was marked increase in healing process as inflammation phase of wound passed quickly. Whereas, when healing efficacy of Lactobacillus bulgaricus and honey treated groups was compared, honey was efficacy of cutaneous wound healing was significantly more than Lactobacillus bulgaricus in terms of wound contraction and subsiding inflammation. It is concluded that holistically use of honey for wound healing in mice is more efficacious therapy than use of Lactobacillus bulgaricus suspension. Availability: Items available for loan: UVAS Library [Call number: 2597-T] (1).
9.
Immuno protective role of newcastle disease virus vaccine (lasota)strainunder immunosuppressive confitions in broilers challanged with ndv isolates of chicken and pigeon orign
by Uqra rauf(2011-va-403) | Dr. Irshad Hussain | Muhammad Aasad ali | dr.Muhammad yasir zahoor.
Material type: Book Publisher: 2017Dissertation note: CD crupt Availability: No items available
10.
Immuno Protective Role Of Newcastle Disease Virus Vaccine (Lasota)Strainunder Immunosuppressive Confitions In Broilers Challanged With Ndv Isolates Of Chicken And Pigeon Orign
by Iqra Rauf (2011-va-403) | Dr. Irshad Hussain | Muhammad Aasad ali | Dr. Muhammad Yasir Zahoor.
Material type: Book Publisher: 2017Dissertation note: CD Corrupt. Availability: Items available for loan: UVAS Library [Call number: 2831-T] (1).