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1. Comparative Efficacy Of Infectious Bursal Disease Vaccines In Broiler Chickens

by Tahir Waheed Khan | Dr. Muhammad Amin Sheikh | Dr. Khushi Muhammad | Dr. Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 1995Dissertation note: The present study was carried out to know the comparative efficacy of different IBD vaccines including Rhone Merleux, Solvay, TAD and CVS in broiler chickens. The evaluation was based on the immune response developed against the vaccine concerned and its assessment through agar gel precipitation test (AGPT) and indirect haemagglutination (IHA) test. All the birds prior to their vaccination were examined for the presence of passive immunity, both through AGPT and IHA. AGPT could only detect 60-80% birds for maternal immunity, whereas IHA showed the presence of this immunity in all the experimental birds. All the vaccinated groups were examined for their immune response on 7th, 14th and 21st day post-vaccination. Both AGPT and H-IA showed the decline in immunity on 7th day post-vaccination and then a gradual increase in titres occurred at 14th day of vaccination. The titres attained their peak on 21st day post-vaccination. With AGPT on 21st day post-vaccination the birds for various vaccines gave precipitin lines respectively as 76, 89, 93 and 100 per cent, for Rhone Merieux, Solvay,TAD and CVS vaccines. With IHA the highest titres obtained on 21st day post- vaccination was 1:128 for each vaccine, however, the number of birds giving this titre varied being only one bird for Rhone Merleux, Solvay , TAD and 3 birds from CVS. Nevertheless, the next best titre obtained was 1:64 and the number of birds giving this titre was 2, Nil, 2 and 8 birds from Rhone Merieux, Solvay, TAD and CVS. The predominant number of birds, vaccinated against each vaccine developed a titre of 1:16 observed at 14th day and 21st day post- vaccination. The challenge infection results showed that birds with lilA titres of upto 1:4 developed severe lesions on their bursae, and other parts of the body, alongwith giving less BBR values. In this way the better IHA titre gave protection to the birds against a challenge infection. All the four vaccines gave identical results for their efficacy against the IBDV infection, however, CVS vaccinated birds developed highest titres in greater number. Availability: Items available for loan: UVAS Library [Call number: 0419,T] (1).

2. Phagocytic Potential Of Chicken Peritoneal Macrophages To Pasteurella Multocida

by Saima Dil | Dr. Muhammad Amin Sheikh | Dr. Khushi Muhammad | Dr. Shakil | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 1995Dissertation note: In this study phagocytic potential/activity of chicken peritoneal macrophages to P. multocida was determined. Macrophages were collected by injecting Sephadex G 75 Sintraperitoneally. Peritoneal exudate cells were collected on 24, 48 and 72 hours post Sephadex G-75 injection. By differential counting macrophages were found to be 70- 80% while the remaining 20-30% cells comprised lymphocytes and heterophils. There was no difference in percentage of macrophages on 24 hours, by single and double dose of Sephadex G-75 but on 72 hours post Sephadexc G-75 injection, percentage of macrophages was a little high by double dose. Both capsulated and non-capsulated forms of P. multocida were chosen for examining phagocytic activity of the chicken peritoneal macrophages and the opsonizing role of the inactivated, normal and hyper immune sera. The capsulated P. multocida was phagocytosed less efficiently in the presence of hyperimmune and : normal sera then the non capsulated organisms. However, as against the latter, the former did not at all respond to the phagocytosing attempts of the macrophages, in the presence of the inactivated serum. Availability: Items available for loan: UVAS Library [Call number: 0449,T] (1).

3. Titration Of Infectious Bursal Disease Virus In Different Organs Of Gumboro Infected Birds And Embryonated Hen

by Saeed Anwar, M | Dr. Muhammad Naeem | Dr. Haji Ahmad | Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 1995Dissertation note: Infectious bursal disease virus (IBDV) was quantitated in different organs of the infected birds and in different parts of the experimentally inoculated embryonated hen eggs through agar gel precipitation technique (AGPT). The AGPT was standardized in the laboratory using known hyperimmune sera against IBDV antigen. The concentration of the agar, antibody and antigen are important factors for the development of precipitation line. Moreover, only gel containing noble agar in distilled water is important for diffusion of the antigen/antibody, while either of the other reagents such as sodium chloride, phenol crystal and sodium azide are presumably added to inhibit the microbial growth. The serum from birds on 14 days post first bost showed 294.1 ACPT titers, while egg yolk from these bird on 30-45 days post first boost were having 68.6 AGPT titers. Postmortem examination showed the lesions In liver,Kidney, spleen, intestine and bursa fabricius. It was observed that only bursa of fabricius showed detectable level (1:4 to 1:16) of IBD virus titers. Similarly, chorioaflantoic membrane of the inoculated egg showed 1:4 AGPT titers. From this study it was concluded that AGPT is a crude but highly reliable test for qualitative and quantitative study of IBDV antigen or antibodies. Availability: Items available for loan: UVAS Library [Call number: 0458,T] (1).

4. A Clinico Pathological Examination And Treatment Of Experimentally Induced Salmonella Infection In Goats

by Baig Muhammad Kakar | Dr. Khalid Pervaiz | Dr. Khushi | Dr. Mohammad Sarwar Khan | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 1997Dissertation note: Salmonellosis is common problem for goat population of the country which is therefore, a constant threat for the farming community of the country during the summer season. The present study was undertaken to study the course of disease with relevance uo environmental factors and the efficacy of various antirnlcroblal drugs available in the market. Salmonellosis was clinically manifested by acute enteritis, dysentery, abdominal cramps, where as temperature, respiration rate and pulse rate were raised upto 2- 3.4°F, 16-23 times/minute and 10-16 times/minute respectively. Predominant postmortem lesions were dehydration, petechiation of the intestinal mucosa occasionally with the accumulation of reddish fluid in t1e intestinal lumen, thickening of the urinary bladder and Pale carcase along with decreased serum sodium concentration in such animals. Among the various antimicrobial drugs used invitro, Gentamycin and chloraphenicol were highly sensitive where as Oxytectracycline, trimethoprim apicillin and kananycin were quite sensitive. Results of in vivo traits for oxytetracycline, tribrissen, gentamycin were 0.00%, 80%, 80% and 100% respectively, whereas 100% mortality was recorded in untreated control group of animals. Availability: Items available for loan: UVAS Library [Call number: 0494,T] (1).

5. Studies Of Antibacterial Activity Of Azadirachata Indica Leaves (Neem)

by Naeem Butt, M | Dr. Muhammed Sabir | dr. Khushi muhammed | Dr. Razzaq Ali.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 1997Dissertation note: One hundred, day old chicks were reared in the college room. Before the introduction of chicks to the shed, the room was washed and disinfected. Azadirachta indica (Neem) leaves were used in the form of powdered, aqueous and methanolic extracts in three major groups i.e. A, B and C (containing 30 birds each) and D with 10 birds, which were kept as a control group. The powdered, aqueous and methanolic extracts were given in the feed to each sub-group at 10,15 and 20mg/Kg body weight to A, B and C i.e., A1, A2 and A3 B1, B2,and B3 C1,C2 and C3 respectively. The N.D. vaccine was administered after 2 days of start of medication. The blood of the experimental and controlled birds were taken at the age of 28, 35 and 42 days. The medication was started at 21 day of age. The sera collected from the blood samples were tested for antibody response against N.D. vaccine. No antibacterial activity of extracts of Neem leaves against Staphylococcus aureous and Pasteurella multocida was observed. the titre of each group at one week post vaccination of ND vaccine to the birds, were determined. It was noted that the water extract of 10mg Neem leaves did not effect the antibody response of birds. The feed conversion ratio of the experimental and control birds were also determined weekly as shown in the Table No.14. There is no apparent different in F.C.R. among the dosage level of powdered, aqueous and methanolic extracts as shown in the Figure-i. During the experimental period ii birds died. The postmortem findings showed E. coli infection. Availability: Items available for loan: UVAS Library [Call number: 0496,T] (1).

6. Standardisation Of Indirect Haemagglutination Test For Monitoring Infectious Bursal Disease Virus

by Sajid Mahmood | Dr. Muhammad Akram Muneer | Dr. Hajid | Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 1997Dissertation note: Indirect haemagglutination (IHA) test was standardized and evaluated to moniter antibodies against infectious bursal disease (IBD). It was observed that oil based vaccine prepared from bursae of fäbricius of infected birds, induced a high level of antibody which were detected by agar gel precipitation test (AGPT). It was recorded Chat tannic acid, glutaraldehyde and chromium chloride had 0.0000781, 0.003906 and 0.0001562 per cent subagglutinating dilutions in normal saline solution (pH 7.2) respectively while 0.000001220, 0.0156 and 0.0025 subagglutinating dilution of the coupling agents were found in phasphate buffered saline (pH 7.2), respectively. Indirect. haemagglutination test is sensitive and specific serological technique to study infectious bursal disease. However, antigen dilution to sensitize erythrocytes, source of erythrocytes, chemical nature of diluent, interaction temperature and time, nature and concentration of coupling agent coated erythrocytes and antiserum against IBD, had influenced the sensitivity of IHA test. Ten percent antigen for sensitizing sheep erythrocytes, incubation temperature of 37°C for 10 minutes for antigen, tannic acid (0.005%) and erythroéyte interaction, freshly prepared sensitized erythrocytes and normal saline solution (pH 7.2) as diluent were found suitable for detecting maximum titre of anti-IBD antibodies through the IHA. Moreover it was observed that the standardized IHA proportionally showed reduction in the titre on dilution of serum. The antibody titre in the IHA was the well having serum dilution, showing resistance to bleed (flow) on tilting the plate for 5 seconds. The final results of antibody titre were achieved within 120 minutes post processing of the samples. Availability: Items available for loan: UVAS Library [Call number: 0529,T] (1).

7. A Clinico Epidemiiological Study Of Bacterial And Parasitic Causes Of Respiratory Syndrome In Sheep And Goats

by Waseem Shahzad | Dr. Khalid Pervaiz | Dr. Khushi | Dr. Muhammad Sarwar Khan | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 1997Dissertation note: In Pakistan sheep and goats production is playing a very important role in bridging the protein gap among our population. Sheep and goats not only provide us high quality meat for our consumption but also wool fibre and high quality leather. In Pakistan every year there is lot of mortality due to respiratory tract infections in sheep and goats especially due to pneumonia which results from a variety of causes' namely, bacterial, viral, parasitic, mycoplasmal and fungal etc. It causes high economic losses due to increased rate of mortality. This study was carried out on two hundred sheep and goats of either sex including all age groups brought to out door hospital of College of Veterinary Sciences, Lahore during February 1997 to June 1997. In the present study isolation and identification of bacterial and parasitic etiological agents of the respiratory tract problems in sheep and goats was carried out. According to this study many bacterial agents causes respiratory problems in sheep and goats, out of 132 positive nasal samples Pasteurella multocida was isolated from 32 (24%), . haemolytica 43 (33%), Corynebacterium 26 (20%), 23 (17%), Streptococcus 18 (14%) and Escherichia ççjj 20 (15%) from affected sheep and goats. Similarly many lungworms cause respiratory problems, out of 82 positive faecal samples Dictvocaulus filaria was isolated from 76 (92.6%) and Protostrongvlus rufesence 6 (7.3%) from affected sheep and goats. The haematological study of the infected and non infected animal showed that there were decreased values of total erythrocytic count, packed cell volume and differential leucocytic count and increased values of total leucocytic count and erythrocytic sedimentation rate of affected animals. Availability: Items available for loan: UVAS Library [Call number: 0539,T] (1).

8. Preparation And Evalution Of Oil Based Egg Drop Syndrome Virus Vaccine

by Ghulam Nabi | Dr. Khushi Muhammad | Dr. Muhammad Akram Munieer | Dr. Shakil | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 1997Dissertation note: The egg drop syndrome virus (Pak-CVS-EDSV: collected from Mirobiology Setion, CVS, Lahore) was culturally characterised by inoculating in embryonated duck eggs, and by haemagglutination (HA) potential of only avian RBC and virus neutralization test. The virus was found antigenically related with the imported vaccinal strain of EDS virus. The virus grew well in the embryonated duck eggs. The HA titer of Allanto-Amniotic fluid (AAF) was more than log 211, while its E1D50 was determined to be 10-10.37 per ml. An oil-based EDSV vaccine was prepared by mixing one part of the AAF with 4 parts of the oil-base. The oil base contained 4% emulsifier (Span-80). The vaccine thus prepared from the local isolate was antigenically comparable with the imported vaccine. The cost of the vaccine production using local strain of the EDS virus was Rs.463/bottle (1000 doses) compared to Rs.1650/bottle of the imported vaccine. The price of the one ml diagnostic antigen was calculated at Rs.20/- compared to Rs.600 per one ml of imported antigen (Market price is Rs.2200/ml of the antigen). Availability: Items available for loan: UVAS Library [Call number: 0541,T] (1).

9. Antibacterial Activities Of An Antibiotic On Complexation With Metals

by Abid Riaz Ahmed | Dr. Muhammad Sabir | Dr. Khushi Muhammad | Mr. Razzaq Ali.

Material type: book Book; Format: print Publisher: 1997Dissertation note: The work presented in this thesis concerns the role of metal ions through chelation in drug design. The area received much attention since the claim that the complexes of drug substances with metal ions are more active and less toxic than the parent drug. In this work complexes of cephalexin with copper and zinc ions were prepared and characterized by microanalysis, IR and UV-vis spectrophotometry, magnetic measurement techniques, differential scanning calorimetry, and atomic absorption spectrometry. The complexes were found to be of ML2 type where M is the metal ion and L stands for the ligand. Both the complexes were monohydrate. On the basis of the analytical and spectroscopic data, the copper complex is suggested to possess an elongatedtetragonal copper (II) ions environment with d22 ground state, the complex was found to be mononuclear with respect to copper atom. The zinc complex possessed square-pyramidal geometry having coordination number 5. The complexes were found to be much more active (copper complex possessed about four times enhanced activity, zinc complex about three times higher activity) against S. aureus. Similarly copper complex possessed 14 times higher activity and zinc complex about 10 times higher activity against E. coli. This work presents a major advance to the pharmaceutical sciences. The complexes reported in this study may replace cephalexin in daily use due to their higher activities against the micro-organisms sensitive to cephalexin. Availability: Items available for loan: UVAS Library [Call number: 0544,T] (1).

10. Detection & Control Of Vaccination (Gumboro Vaccine Nobilis Strain D-78 Stress Against Infectious Bursal Disease

by Subtain, Syed M | Dr. Shakil Akhtar Khan | Dr . Asim Aslam | Dr . Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2000Dissertation note: This research programme was intended to ascertain the stress produced after vaccination with live intermediate type of infectious bursal disease virus (IBDV) vaccine, which was administered orally through drinking water. It was also intended to manage the effects of vaccination stress with the supplementation of vitamins and aspirin. One hundred and sixty day old layer chicks were divided into four experimental groups i.e. A, B, C and D, 40 birds in each group. Group A was kept as control (non-vaccinated), B was given vaccine but not medicated, group C was administered vaccine as well as multivitamins for 3 days post-vaccination while group D was also medicated with aspirin for 3 days post-vaccination. The studied parameters were: heterophil/lymphocyte ratio, serum biochemical analysis (serum protein, glucose and cholesterol), determination of antibody response against IBDV. At the end of experiment (42nd day) adrenal glands were isolated from 10 randomly selected birds from each group. The glands were subjected to gross and histopathological Availability: Items available for loan: UVAS Library [Call number: 0678,T] (1).

11. Passive Immuniation Of Newcastle Disease Virus Infected Birds

by Raheel Arshad | Dr . Khushi Muhammad | Dr . Khalid | Dr . Sa meera Akhtar | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2000Dissertation note: Present study was designed to determine the contribution of maternal immunity against the Newcastle disease (ND). In the study two hundred and ten white leghorn cockerels were used. A velogenic ND virus field isolate was used for the challenge. Lethal Dose 50 (LD5O) of the velogenic strain of ND virus was calculated which was 10.727. Maternal antibody levels of the chicks were determined on weekly basis by using the haemaggldtination inhibition (HI) test. The maternal geometric mean HI titre recorded at 1, 7, 14, 21 and 28 days of the age of birds were 147, 157.6, 22.6, 13 and 2.6 respectively. The protection offered by maternal immunity in chicks challenged with 100 LD50 at 1, 7, 14, 21 and 28 days of age was, 100, 100, 70, 10 and 0 percent respectively. It was concluded that birds having maternal hurnoral immunity more than 64 GMT showed 100% protection to challenge infection. The effect of immunized egg yolk to ND infected birds was also studied. The HI titre of immunized yolk was determined and then different HI units of the immunized yolk were prepared with normal saline. It was observed that egg yolk (1 ml, 64 HI units) injected to ND challenged birds showed 100% protection as compared to that of control group (given 1 ml of 0 HI units of the yolk). The cost of the production of immunized yolk was also determined that was Rs.0.35 / dose (1 ml : 64 HI units). From the study, it was inferred that the hyper immunized yolk can be used as therapeutic agent to cure the ND infected birds. Availability: Items available for loan: UVAS Library [Call number: 0679,T] (1).

12. Immuno Prophylaxis Of Entrerotoxaemia In Sheep And Goats

by Shahzad Jawed | Dr . Muhammad Naeem | Dr . Asif | Dr . Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 1999Dissertation note: The present project was designed to study the comparative efficacy of three different adjuvant (Potassium aluminium sulphate, Aluminium hydroxide gel and mineral oil) for enhancing the immune potential of enterotoxaemia (combined) vaccine. T)ifferent standard biological media alongwith the supplementation of amino acids, minerals, plants and animal extracts and special modified formulated media were used for the production of alam precipitated vaccine, aluminium hydroxide gel adsorbed vaccine and oil adjuvant vaccine. It was concluded that addition of various ingredients, including yeast extracts, trace elements, amino acids, plants and animal extract, and cystine hydrochloride in proper concentration, increased the level of prototoxin and toxin in culture media due to the availability of essential required nutrients. Mouse model was chosen to study the safety and potency test of all the vaccines. The potency of all three vaccines was compared. In this experiment alam precipitated vaccine proved inferior to aluminium hydroxide gel adsorbed and oil adjuvant vaccine. In case of oil based vaccine especially in sheep the IHA antibody was significant in vaccine having potency of 750 HU/ml but on the other hand vaccine have potency 250 and 500 HU/ml were proved non significant, and the day 45 was proved significant in developing the antibody titre than that of 15 and 30 days. Same the picture was observed in case of goals that oil adjuvant vaccine was significant that of aluminized and toxoid adsorbed vaccine. It was observed that the protection afforded to goats by multivalent clostridial vaccine was higher than afforded to sheep. Availability: Items available for loan: UVAS Library [Call number: 0683,T] (1).

13. Passive Immunization Of Infectious Bursal Disease Infected Broiler Chicks

by Fazli Rabbi | Dr . Khushi Muhammad | Dr . Khalid | Dr . Muhammad Akram Munir | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2000Dissertation note: Infectious bursal disease (IBD) is a common problem in commercial unvaccinated birds and causing heavy economical losses to the poultry industry. Chicken layers when primed with oil based 113D vaccine at age of 13 weeks and boosted with the same vaccine at 15 weeks of age showed high titre of yolk agar gel precipitating (AGPT) antibodies against IBD virus when tested on 21 and 28 weeks of age. Storage temperature (+4°C and -20°C) had undetectable effects on the physical properties (color and smell) and AGPT titres of the hyperimmunized yolk contnining 0.5% forrnalin (v/v). r[he AGPT antibody titre of the hyperimmunized yolk had good correlation with the enzyme linked immunosorbant assay (ELTSA) titre of 113D virus antibodies (r: 0.92). The IBD infected broilers (28 days old) when passively immunized with the yolk (one ml: 64 AGPT units of IBD antibody titre) induced 80% recovery as compared to that of untreated (control) birds. It is anticipated that the hyperimmunized yolk may he used as a therapeutic agent to cure the IBD infected birds. Availability: Items available for loan: UVAS Library [Call number: 0687,T] (1).

14. Comparative Efficacy Of The Latest Antibacterial Drugs Against Experimentally Induced Pullorum Disease

by Dr . Khalid Omran, M | Dr . Khalid Pervaiz | Dr . Khushi | Dr . Mohammad Sarwar Khan | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 1997Dissertation note: A total 210 birds were reared upto the age of 4 weeks and the divided into 7 groups viz A, B, C, D, E, F and G, comprised of 30 birds in each group. At the age of 28 days the groups A to E were experimentally infected with Salmonella pullorum inoculation intraperitoneally with the dose rate of '/2 ml. The group F was infected but non-medicated and the group G was kept as control (non infected and non medicated). The groups A, B, C, D, E and E were treated with Anflox, Inoxyl, Triquine, Flumiquine and Gentamycine respectively. All the groups were kept under close observation to record clinical signs, mortality rate, weight gain and feed conversion ratio. Postmortem of dead birds was also conducted. The blood parameters TEC, TLC, and DLC were also estimated post medication on 1st, 3rd and 5th day. The mortality before medication in groups A, B, C, D, E and F was 10%, 16.67, 6.67%, 13.34%, 13.34% and 6.67% respectively. While the mortality during treatment in each group was 25.92%, 32%, 39.28%, 19.23%, 46.16%, 64.28% respectively. The mortality in control group was 6.66% during these days. According to this trial Flumiquine provided maximum protection against Salmonella pullorum infection and proved best in relation to weight gain and FCR. Anflox stood second in the list while Inoxyl was the 3rd drug which provided protection against the infection, whereas Triquine and Gentamycin were the least effective drugs. The signs which appeared after 18-24 hours post infection were listlessness, ruffled feathers, droopy wings, loss of appetite, poor growth, depression, increased thirst and severe diarrhoea of chalky white color. The post mortem findings were enlarged spleen, congested liver with streaked haemorrhages, congested and distended kidneys, grey hepatization in lungs and enlarged heart. The total erythrocytic count decreased in infected non-medicated birds while in medicated groups it remained in normal range. The same was true of total leucocytic count. However, in DLC the heterophil indicated increased percentage after inoculation of infection while the lymphocytes, basophils, eosinophils and monocytes remained within range after the infection. The findings of the present study elucidate the disease and help in diagnosis and treatment of this malady. Availability: Items available for loan: UVAS Library [Call number: 0688,T] (1).

15. Preparation & Evaluation Of Alum Precipitated & Oil Based Haemorrhagic Septicaemia Vaccines

by Dr . Khushi Muhammad Zulfiqar Ali | Dr . Khushi Muhammad | Dr . Asif | Dr . Lrshad Hussain | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 1998Dissertation note: Pasteurcila multocida was isolated and characterised on the basis of cultural biochemical serological and pathogenicitytests.The dense culture of the organism was achieved in a fermenter that was provided sterilized air during incubation. Two types of the formalin inactivated Pastcurclla multocida vaccines (oil-based and alum precipitated) were prepared and their efficacy was evaluated in bovine. It was observed that oil-based haemorrhagic septicacmia (HS) vaccine induced high level of indirect haemalutiiiating (IHA) antibodies in the vaccinated cattle which persisted for more than 6 months. In contrast, alum precipitated HS vaccine induced immunity breakdown in the cattle with high titres of IHA antibodies while induced mw level of IHA antibodies, which persisted for 4 months. Availability: Items available for loan: UVAS Library [Call number: 0694,T] (1).

16. Studies Of The Production And Detection Of Haemolytic Toxin In In Vitro Culture Of Clostridium Perfringens Type D

by Bakht Sultan | Dr . Khushi Muhammad | Dr . Haji Ahmad | Dr . Sameera Akhtar | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 1998Dissertation note: Physicochemical factors modulating the production of haemolysin in culture of Clostridium perfringens (type-D) were evaluated. It was observed that toxin was produced in all the three media. The maximum titer of (4184) was achieved in RCM. The titer in thioglycollate was 2088 and in RCM with K2HPO4 were 1248 after 24 hours incubation. It was observed that pH 6.5, 7.0 and 7.5 of the medium before incubation resulted 1024, 4184 and 1576 haemolytic titers. Anaerobic environment and neutral pH during incubation augmented the haemolysin production in the culture. Trypsin 0.1 percent in the culture filtrate converted the prototoxin into haemolysin which exhibited maximum lytic activity in 60 minutes interaction time. Trypsin solution (1 percent) alone failed to induce haemolysis while the haemolysin showed maximum haemolytic activity at 37°C. The trypsinised culture supernatant (haemolysin) induced lysis of erythrocytes of sheep, goat, horse and chicken. The resultant high titer of haemolysin unveiled the propects of preparation of combined vaccines for sheep and goats. Availability: Items available for loan: UVAS Library [Call number: 0698,T] (1).

17. Preparation And Evaluation Of Newcastle Disease Virus (Mesogenic Strain) Oil Based Vaccine

by Shafi Ullah Chand | Dr . Khushi Muhammad | Dr . Sameera Akhtar | Dr . Shakil | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 1998Dissertation note: The present work was proposed to prepare oil based Newcastle disease virus vaccine and to compare its efficacy with imported vaccines. An oil based ND vaccine was prepared using moderately virulent strain of NDV. The virus was cultured in chicken embryos. The allanto-amniotic fluid, chorioallantoic membrane and infected embryo (virus suspension) was subjected to titration. The HA titer of allanto-amniotic fluid (AAF), chorloallantoic membrane (CAM) and embryo was upto 512, 1024 and 2048, respectively. The MD50 was calculated to be 1088/0.1ml. Effect of temperature on its keeping quality was determined by estimation of its HA potential at various intervals. The AAF was processed for inactivation, sterility and safety tests. Formalin at a rate of 0.12% inactivated the NDV in 48 hours at 37°C. Addition of antibiotic such as gentarnycin and nystatin inhibited common contaminants. An oil based NDV vaccine was prepared by mixing one part of processed AAF in 4 parts of oil base. The oil base contained 4% emulsifier span-80 and 1% tween-80. The vaccine thus prepared from moderately virulent strain was antigenically comparable with the imported ND vaccine. The cost of vaccine production using moderately virulent NDV was Rs.463/bottle (1000 doses) compared to RS.1250/- per bottle of imported vaccine. The price of one ml diagnostic antigen was calculated at Rs.2/ml. The results of present project encourage to develop an economical and effective oil based ND vaccine and diagnostic HA NDV antigen. Availability: Items available for loan: UVAS Library [Call number: 0700,T] (1).

18. A Study On Physico Chemical Actors Affecting The Survial Of Avian Influenza (H9 N2) Virus

by Tehmina Sadaf | Dr . Sameera Akhtar | Dr . Khushi Muhammad | Dr . Shakil | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2001Dissertation note: A total of 310 chicken embryos (9 day old) were purchased from local market. The embryos were incubated at 37°C. On eleventh day of age avian influenza virus (H9 N2) for its propagation was inoculated in 10 chicken embryos. The four haemagglutination titer (4HA) of the virus was prepared to observe the response to various physical and chemical factors. Physical factors included were temperature, pH and UV (280 nm) light. The virus, exposed to the physical factors for different time intervals was inoculated into embryos through allantoic route. These embryos were kept in an incubator (37°C) for 72 hours. Later on the allantoic amniotic fluid (AAF) from each inoculated embryos subjected to spot haemagglutination test. The virus endured 56 °C temperature for 15 and 30 minutes while got inactivated in 45 minutes. More over virus survived at pH 7 for 15, 30 and 45 minutes but lost its HA activity at pH 5 and 9 in 15, 30 and 45 minutes. It was further examined that virus survived after 60 minutes exposure UV light but inactivated after 90 minutes. The disinfectants formalin, phenol, iodine solution and fin virus were used in 0.5%, 1% and 1.5% concentrations. The 4HA titer of virus was mixed in various concentrations of chemical disinfectants and was inoculated into embryonated eggs. The AAF of these eggs was subjected to spot agglutination test. The results of the test showed that all four chemicals formalin, phenol, iodine and fin virus inactivated the virus in 0.5% concentration in 15 minutes and all have good antiviral activity against avian influenza virus. Availability: Items available for loan: UVAS Library [Call number: 0703,T] (1).

19. Efficacy Of Various Chemical Agents Against Avian Influenza Virus (Type H9)

by Imran Altaf | Prof.Dr.Muhammad Ashraf | Dr. Khushi | Dr. Muhamad ovais Omer.

Material type: book Book; Format: print Publisher: 2001Dissertation note: In the present project, 300 eggs were taken from Big Birds. They were cleaned and incubated for 9 days. After 9 days they were candled to separate the live and dead embryo. On the 11th day, the embryos were inoculated with the different concentrations of drugs which were to be tested. The detail of these drugs are given below: Chlorine was taken in the concentration of 1, 2 and 5 ppm. They were divided into control and experimental groups containing drug + Normal saline and drug. + virus. The results showed that 2 ppm and 5 ppm chlorine was most effective as disinfectant, while 1 ppm was not effective. Iodine was also tested in the concentration of 0.1, 0.5 and 1 per cent for control and experimental groups in the same fashion as in chlorine. The results showed that 1% and 0.5% solutions are highly antiviral but are toxic for living cells, while 0.1% iodine is virucidal against virus and can be used for the living cells as it was not toxic for embryo. In case of amantadine 50 jig/mi, 500 jig/mi and 1000 jig/mi were used along with virus and Normal saline for experimental and control groups, respectively. Results show that 1000 i.g/ml concentration was toxic for cell and hence can not be used in vivo, while 50 jig/ml failed to stop the replication of virus. 500 jig/ml concentration not only stopped the viral replication but also did not kill the embryo. So 500 jig/mi of amantadine can be used in poultry, if desired. Acyclovir is another pharmaceutical product, used in the concentration of 50 jig/mi, 200 jig/mi and 400 jig/mi in the fashion as described above. Results showed that acyclovir was effective within the range of 200 jig/ml to 400 jig/mi without any fear of damage to cell, but 50 jig/mi concentration failed to stop the viral replication and also showed high HA titer. In herbal group, Soyabean was tested in the concentration of 1 g/l00ml, 5g/l00ml and 10 g/l00ml. When the results were collected it was noticed that neither concentration of soyabean has any antiviral effect against influenza. Opuntia plant group which comprises of three species named: 0-Stricta, O-Dellinii and 0-Manocantha was testified for their antiviral efficacy. The concentrations used were 1 gm, 5 gms and 10 gms dissolved in 100 ml of Normal saline. Their extracts were obtained and injected in the embryo alongwith virus and Normal saline for experimental and control groups after giving them an interaction time of one hour. In the results, 10 g/l00ml and 5 g/l00ml show a good anti-viral efficacy against influenza, while 1 g/l00ml failed to show any significant effect and gave high HA titer. So in Pakistan these herbal drugs should be further investigated so that these can be used against influenza. Availability: Items available for loan: UVAS Library [Call number: 0743,T] (1).

20. Imunization Of Rabbits Against Boophilus Microplus Using Midgut And Salivary Glands

by Mohammad Asif | Dr. Khalid Saeed | Dr. H. A. Hashmi | Dr. Khushi | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2002Dissertation note: The current research was conducted as a simulation model in rabbits for the development of vaccines against cattle ticks Boophilus micro/us using tick salivary glands & midgut as the sources of antigens. The comparative efficacy of these vaccines was evaluated on the basis of antibody titres and tick rejection on the host in terms of mortality, decrease in egg laying and decrease in hatchability of eggs. It was found that the vaccine prepared from the midgut of B. microplus gave better results than the vaccine prepared from salivary glands. The highest percentage of dead ticks was observed in animals which were given midgut vaccine (21.82%). This vaccine also caused a significant decrease in the egg laying capacity of the ticks (35.83%). However, none of the vaccines had any significant effect on the hatchability of eggs laid by the ticks. The highest antibody titers were observed in rabbits injected with midgut vaccine (GMT=5.50), which was 45.28% higher than rabbits given salivary gland vaccine. The overall success rate for midgut vaccine, in terms of tick mortality on host, decrease in egg laying capacity and antibody titer,. was 37.06% for midgut vaccine while only 16.51% for salivary gland vaccine. The results are very encouraging and it is hoped that with the original host i.e. cattle, more satisfactory results can be achieved. Availability: Items available for loan: UVAS Library [Call number: 0771,T] (1).

21. Passive Immunization Of Avian Influenza Virus Infected Broiler Chicks

by Muhamad Mahmood Mukhtar | Dr. Masood RAbbani | Dr. Khushi Muhammad | Dr. Shakil | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2002Dissertation note: Present study was conducted to determine the contribution of passive immunization against avian influenza disease. In this study, ten layer birds were vaccinated thrice at 21 days interval, using oil based avian influenza v irus (AIV: H7 type) vaccine. A high titer of anti-Al V-antibodies in blood serum and egg yolk of these birds was determined on 20(11 day post-boosting using haemagglutination inhibition (HI) test. A virulent avian influenza virus (H7 type) with mean embryo infective dose (E1D50) of l0 was used for challenging the birds. The hyperimmune serum and hyperimmune egg yolk (1ml, 128 HI units) injected to avian influenza virus (H7 type) challenged broiler chicks showed 100% protection as compared to that of virus control group (given I ml of 0 HI units of serum and yolk). Moreover, it was found suitable to passively immunize the birds before exposure or simultaneously with the exposure of avian influenza virus. The cost of the production of hyperimmune egg yolk was calculated as Rs. 0.43 per dose (1ml: 128 HI units), which was quite economical as compared to other chemotherapies. It is concluded that the hyperimmune serum and hyperimmune yolk can therapeutically be used to cure the avian influenza virus (H7 type) infected birds. Availability: Items available for loan: UVAS Library [Call number: 0776,T] (1).

22. Passive Immunization Against Hydropericardium Syndrome Infected Broilers

by Ghazanfar khalid | Dr. Khushi Muhamad | Dr. Masood Rabbani | Dr. Shakeel | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2002Dissertation note: In this project, passive immunization against hydropericardium syndrome virus (HPSV) infected broiler chicks was studied. The hyperimmune yolk and serum were raised in commercial layers by priming and boosting with formaldehyde inactivated HPSV vaccine. It was found that yolk and serum collected from the layers showed high titres of indirect haemagglutination (IHA) antibodies against HPS virus. Lethal dose 50 (LD50) of the HPS virus infected liver homogenate was calculated to be 1O-55/ml. It was noted that the broiler chicks (26 days old) receiving yolk containing 256, 128, 64 and 32 units of IHA-anti-HPSV antibodies and virulent HPSV, simultaneously showed 100%, 100%, 100% and 60% protection. While the broiler chicks receiving serum containing 128, 64, 32 and 16 units of IHA anti-HPSV antibodies and virulent HPSV, simultaneously showed 100%, 100%, 40% and zero protection. The birds receiving yolk and serum of control group showed zero protection. It was observed that egg yolk (lml, 64 IHA-anti HPSV-antibodies) injected 24 hours before, at the same time and after 24 hours to HPSV challenged broilers showed 100% protection. While the clinically healthy birds 48 hours post challenge infection showed 60% protection and birds showing signs of the disease showed 20% protection. The cost of the production of hyperimmune yolk was Rs. 0.50/dose (lml: 64 IHA units of anti-HPSV antibodies). It was concluded that hyperimmune yolk could be used to cure the HPSV infected birds if administered immediately in the affected flocks. Availability: Items available for loan: UVAS Library [Call number: 0781,T] (1).

23. Comparative Immunogenicity Of Different Hydropericardium Syndrome (Hps) Vaccines In Broiolers

by Arfan Ahmad | Dr. Sameera Akhtar | Dr. Khushi Muhammad | Dr. Shakeel | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2002Dissertation note: Formaldehyde inactivated hydro-pericardium syndrome (HPS) vaccine without any adjuvant (F-HPS), an oil based HPS vaccine (0- HPS) and alum precipitated HPS vaccine (A-HPS) were prepared and comparative immunogenicity was evaluated in broilers. These vaccines were injected to each bird of groupG1, 02 and G3 sub-cutaneously on 14t day of their age, respectively while the birds of group G4 were kept non-HPS vaccinated control. Each of the bird of each group was also vaccinated against Newcastle disease virus-NDV (LaSota strain: eye droppings) while birds of group G5 served as ND non vaccinated control. Each of the vaccine induced detectable level of anti-HPS virus indirect haemagglutination (IHA) antibody titre. The 0-HPS vaccine induced higher titre ofthe anti-HPSV-IHA antibody titre that of F-HPS and A-HPS vaccines. All of the three vaccines induced resistance in the birds that showed 100% protection when were given challenge infection on 14 days post-vaccination while the birds of control group showed zero percent protection. At the time of challenge infection, anti- Newcastle disease virus haemagglutination inhibition (ND V-HI) antibody titres were same in the HPS vaccinated and un-vaccinated broilers. It is concluded that all the vaccines induced effective immunity in the birds. The 0-HPS vaccine induced higher levels of anti-HPS virus IHA antibody titres than that of F-HPS and A-HPS vaccines. Moreover, non of the vaccine induced detectable level of immuno-modulatory effect on the anti-NDV-HI antibody titre of birds to NDV vaccine. Availability: Items available for loan: UVAS Library [Call number: 0787,T] (1).

24. Detection And Control Of Vaccination Stress Following Vaccination With Live Virus Newcastle Disease Vaccine And Its Effect on Immune Response Commmercial Broiler Chicks

by Zahid Jawad | Dr. Shakil Akhtar Khan | Dr. Khushi Muhamma | Dr.Javed Rashid | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 1997Dissertation note: This study intended to ascertain the vaccination stress following live Newcastle disease vaccine by oral route and to determine the comparative efficacy of probiotic and vitamins to combat it. One hundred and forty day-old broiler chicks were divided into four groups i.e. A, B, C and D having 35 birds each. The birds were kept for 45 days alter vaccination with Bio-LaSota by oral route on 21st day. Chicks From group A was kept as control. Group B was given vaccine and no treatment. Group C was given vaccine and only brobiotic (protexin). Group D was given vaccine and only vitamins (Vitaoligosol). Seven birds of each group were slaughtered on 22nd day, 23th, 24th and 25th day and remaining 7 birds of each group were slaughtered on day 45th, for collection of blood samples. The blood samples from 7 randomly selected birds were collected on day 16, 30th and day 45th for determination of antibody t.itre. The following parameters were studied: (i) determination of hetrophil/lymphocyte ratio (Ii) estimation of antibody response against Newcastle disease vaccine (iii) estimation of serum biochemical substances (iv) Determination of adrenal gland body weight index and (v) Pathological study of adrenal glands. Live virus vaccine against Newcastle disease caused vaccination stress in broiler chickcns. The birds expressed vaccination stress for a variable period ranged from 1 to 2 post-vaccination day. But these results were more accurate if level of corticosteroids was detected. In future, a more comprehensive study is required to devise simple and reliable methods for detecting stress in poultry in field conditions and also to suggest therapeutic and prophylactic measures for relieving the birds from the state of stress. Availability: Items available for loan: UVAS Library [Call number: 0806,T] (1).

25. Passive Immunistion Of Pasteurella Multocida Infected Rabbits

by Ali Ahmad | Dr. Masood Rabbani, Asso.Prof., CDL | Dr. Khushi Muhammad, Associate Prof | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2003Dissertation note: Haemorrhagic septicaemia (HS), an important bacterial disease of buffaloes and cattle, is caused by Pasteurella multocida. Improved management practices and regular vaccination programme have significantly contributed to lowering the incidence of the HS disease in our country. The outbreaks are mostly experienced in young animals, especially, calves (Sheikh et al., 1996). Present study was conducted to determine the contribution of passive immunization against HS in infected rabbits. In this study, 5 rabbits were vaccinated thrice at 21 days interval, using oil base haemorrhagic septicaemia vaccine (OBHSV). A high titre of anti-Pasteurella multocida-antibodies in blood serum was determined on 56th day post boosting using indirect haemagglutination (IHA) test. A virulent Pasteurella multocida with mean lethal dose (LD50) of 10-6.749 was used for challenging the rabbits. The hyperimmune serum (1 ml, 256 IHA units) injected intravenously to Pasteurella multocida challenged rabbit showed 100% protection as compared that of intramuscularly injected serum, which showed 66.66% protection. Similarly the antigen control group showed 0% protection. Moreover, it was found suitable to passively immunize the animals before exposure or simultaneously with the exposure of Pasteurella multocida. It is concluded that the hyperimmune serum can therapeutically be used to cure the Pasteurella multocida infected rabbits. Availability: Items available for loan: UVAS Library [Call number: 0816,T] (1).

26. Passive Immunization Against Canine Parvovirus In Dogs

by Umer Ahmad | Dr. Masood Rabbani | Dr. Asim Khalid | Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2003Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 0836,T] (1).

27. Studies On Teh Physicochemical Factors Affecting Keeping Quality Of Hyperimmune Yolk

by Jawad Nazir | Dr. Khushi Muhammad | Dr. Kamran | Dr. Masood Rabbani | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2003Dissertation note: Present study was conducted to investigate the production of immune yolk against multiple avian viruses and effect of physicochemical factors on its keeping quality. It was observed that peak antibody titers in the yolk of eggs laid by the birds vaccinated against avian viruses (Newcastle disease virus-NDV, infectious bursal disease virus-IBDV, avian influenza virus-AIV-H9 and hydro pericardium syndrome virus-HPSV) were attained on 4 weeks post-boosting which were maintained over subsequent 6 weeks and started declining thereafter. The immune yolk treated with chemicals (antibiotics, sodium azide and formaldehyde) was stored at room temperature, refrigerator and freezer. Any change in physical properties (color and odor) and antibody titer of the yolk was determined at day 0, 7, 14, 22 and 30 post-storage. Antibiotics (penicillin, streptomycin and gentamycin) in the yolk during storage at room temperature inhibited the bacterial growth but permitted fungal growth that induced physico-chemical changes such as change in color, development of bad smell and reduction in antibody titer. Antibiotics / sodium azide treatment and freezing / refrigeration for more than 30 days showed undetectable change in antibody titer of the immune yolk. However, formaldehyde in the yolk during storage at -20°C precipitated its proteins leaving clear fluid free from the antibodies. Effect of chemically treated stored immune yolk was investigated on the recovery of Newcastle disease virus (NDV) infected layer cockerels (35 days old). Antibiotics and sodium azide treated fresh and stored immune yolk (at 4°C for 15 days) containing 64 units of anti-ND V-haemagglutination inhibition (HI) antibodies showed 100 percent protection in the birds. The immune yolk treated with the same chemicals and stored at -20°C for 30 days also showed 100 percent protection. However, antibiotics and sodium azide treated yolk (containing same titer of the antibodies) stored at 4°C for 30 days showed 70 percent and 90 percent protection, respectively. It is inferred that sodium azide in the immune yolk during storage at 4°C or -20°C might have preserved antibodies and hence such yolk may be used for passive immunization to treat the virus infected birds. Availability: Items available for loan: UVAS Library [Call number: 0840,T] (1).

28. Antibody Response Of Buffalo To Inactivated Foot And Mouth Disease (Aphtho) Virus Vaccine

by Nadeem Murtaza | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2005Dissertation note: Buffaloes when vaccinated against Foot and Mouth Disease (FMD) vaccine containing serotype "O" of the virus, showed detectable level of complement fixing (CF) antibodies. Buffaloes vaccinated with Aftovaxpur vaccine showed undetectable level of CF-antibodies when tested through complement fixation test using local vaccinal serotype "O" of FMD virus. However, buffaloes irrespective of age and sex when vaccinated with aluminum hydroxide adsorbed FMD vaccine containing serotype "O" of the FMD virus, showed detectable level of CF-antibodies, when tested through CFT using the local serotype "O" Of FMD virus. These antibodies disappeared fourth month post boosting. Buffalo calves immunized with oil based FMD vaccine showed high-level GMT titer (17.6) of the CF-antibodies. Rabbits immunized with the oil based FMD vaccine showed high level GMT (31.2) of the CF-antibodies. Low level of CF-antibodies might be sufficient to induce resistant to field exposure of the FMD virus serotype in the presence of blood complement. Sera of buffalo, cattle, sheep, goat and guinea pigs contained complement titer 35.2, 32.6, 19.2, 20.8, 614.4, respectively. Moreover, it was observed that complement activity remained stable when stored at -200C for 24 hours. The complement activity decreased from 1:512 to 192, 70.4, and 13.6 when stored at 40C, 250C and 370C, respectively. The complement activity was detectable when diluents containing Ca++and Mg++ ions were used. Availability: Items available for loan: UVAS Library [Call number: 0877,T] (1).

29. Development Of Standard Protocols For Preparation And Evaluation Of Liver Homogenate Vaccines Against

by Sahidullah | prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: Twelve vaccines were prepared from HPS infected liver homogenate by using two different virus concentrations (1x102 &1x103 LD50) and two virus inactivants (1%formalin and 0.001%Binary ethyleneimine) with and with out different adjuvants. These vaccines were evaluated in 13 groups of day old broilers (105 chicks) for their comparative immunogenicity and protection. At day 14 of age, groups A1, B1, C1 & D1 were vaccinated with 4 oil base vaccines (OB-HPSV) with different virus concentration and different inactivants. Similarly groups A2, B2, C2 & D2 were vaccinated with aluminized vaccines (AH-HPSV) and groups A3, B3, C3 & D3 with non adjuvant vaccines (NA-HPSV). Groups E was kept as unvaccinated control group. All the vaccinated birds were found sero-positive 7 days post vaccination (PV). IHA GMT results indicated no difference for different virus concentrations and different virus inactivants but same adjuvant. The IHA GMT recorded weekly during 0-28 days post vaccination was the highest and more consistent (52-181) for OB-HPSV followed by AH-HPSV (52-147) and then NA-HPSV (73.3-104). All the birds vaccinated with OB-HPSV resisted the virus challenge 21 days PV (100% protection). While protection percentage recorded for AH-HPSV and NA-HPSV was 87.5 % & 62.5% respectively. It was concluded that 1x102 LD50 oil base vaccines inactivated with either formalin or binary amine may be recommended for commercial use being the best in experimental trails. Availability: Items available for loan: UVAS Library [Call number: 0878,T] (1).

30. Standardization Of Indirect Enzyme Linked Immunosorbant Assay For Detection Of Antibodies Against Foot and Mouth Disease Virus Sdrotype "O"

by Yasmeen Siddique | Prof. Dr. Khushi Muhammad | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2005Dissertation note: An indirect ELISA was standardized for titration of anti-FMD-serotype-specific antibodies. In this test polystyrene plates were coated with known FMD serotype "O" virus using carbonate/bicarbonate buffer. The blank spaces were blocked with horse serum. The immunoplate was coated with anti-FMD "O" virus specific serum from vaccinated calves. After washing, the plate was coated with rabbit anti-bovine-Ig-specific-antibodies-horse radish peroxidase conjugate. After washing, the plate was coated with HRP specific substrate. Development of color was recorded in form of OD value using ELISA reader. During the standardization of ELISA, flat bottom ELISA plates among all types of plates, 1:10 diluted virus among different dilutions of FMD "O" type virus, 1:10 diluted serum from buffalo calves vaccinated with FMD "O" type specific vaccine, 1:4000 dilution of conjugate and incubation of 4º C for coating the virus showed good results. In each experiment, plateau region, test back ground and plate back ground was recorded. Results of the study will help in establishment of an economical, sensitive, reliable indirect ELISA that subsequently be helpful to understand the percent prevalence of FMD serotypes in Pakistan and efficacy of FMD vaccines. Availability: Items available for loan: UVAS Library [Call number: 0879,T] (1).

31. Propagation Of Hydropericrdium Syndrome Virus In Laboratory Host Systems

by Basharat Mahmood | Dr. Mansur-ud-Din Ahmad | Dr. Azhar | Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: Hydropericardium syndrome primarily affects the broilers between the age of 2-7 weeks. A vaccine prepared from infected liver extract treated with formaldehyde is being used to protect the broilers from the disease. The current study was carried out to propagate the hydropericardium syndrome virus in various laboratory host systems i.e. embryonated hen eggs, primary chicken embryo liver (CEL) cells, chicken embryo kidney (CKC) cells, chicken embryo (CEF) fibroblasts and BHK 21 cell line. The protocol for cultivation of primary chicken embryo liver cells, chicken embryo fibroblasts, chicken embryo kidney cells were standardized under local conditions. Liver samples were collected from HPS affected birds were processed and propagated in embryonated hen eggs, chicken embryo liver cells, chicken embryo kidney cells, chicken embryo fibroblasts and BHK-21 cell line. The comparative sensitivity to hydropericardium syndrome virus was studied. Five field samples were recovered out of seven after reproducing the disease in susceptible healthy broilers. These five liver samples were propagated in all laboratory host systems. It was recorded that hydropericardium syndrome virus could be propagated in chicken embryo liver cells and chicken embryo kidney cells. Hydropericardium syndrome virus could not be detected in AAF of embryonated hen eggs inoculated through allantoic, chicken embryo fibroblasts and BHK-21 cell line. Microtitration technique was used to determine the titer of the propagated virus. The serological techniques used to confirm the presence of HPS virus in cell culture supernatant and the allento- amniotic fluid were AGP test and serum neutralisation. Polyclonal antisera was raised using formalized liver homogenate vaccine and oil- based cell culture propagated vaccine. Polyclonal antiserum against HPS virus using oil-based cell culture propagated vaccine was found to Specific against liver homogenate collected from HPS affected birds. Availability: Items available for loan: UVAS Library [Call number: 0880,T] (1).

32. Passive Immunization Against Canine Distermper Virus In Dogs

by Ali Ahmed Malik | Prof. Dr. Masood Babbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: Canine distemper is an important, highly contagious disease of dogs, caused by morbillivirus of family paramyxoviridae. The disease occurs worldwide in variety of hosts. In the present study, data relative to breed, sex and age susceptibility in clinically suspected cases of canine distemper was collected and analyzed. The disease is mostly seen in young nonvaccinated dogs of 4 to 6 months of age when maternal anti-CDV antibodies are decreased. Immune serum was raised in experimental dogs with commercially available measles live virus vaccine. The level of antibodies in the immune serum was determined by agar gel precipitation test (AGPT) and an ELISA based assay. Immune serum containing 128 AGPT units of anti-CDV antibodies was effective to control the disease in infected dogs after natural exposure to canine distemper virus. Finally the effective time for passive immunization against canine distemper was determined in experimental dogs. It was noted that immune serum offered protection to canine distemper immediately after infection, during the incubation period of the disease , 48 hours after infection and early phase of the disease(at the appearance of clinical signs). Passive immunization is not rewarding in the terminal phase of the disease (when infected dogs show nervous signs of the disease).Thus it is very useful for the prevention of disease in dogs kept with infected dogs in kennels and pet shops. Availability: Items available for loan: UVAS Library [Call number: 0882,T] (1).

33. Standardization Of Indirect Sandwich Enzyme Linked Immunosorbent Assay For Detection Of Foot And Mouth Disease

by Muhammad Mujahid Amjed | Prof. Dr. Khushi Muhammad | Professor Dr. Masood Rabbani | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: Foot and Mouth Disease (FMD) is one of the most troublesome and infectious diseases of livestock, caused by the FMD virus. In this study Indirect Sandwich Enzyme Linked Immunosorbent Assay (IS-ELISA) was standardized to characterize the FMD serotype "0" virus. Oil based FMD serotype "0" vaccine was prepared and injected at the neck region of guinea pigs and rabbits. The vaccine induced anti-FMD serotype "0" virus antibodies in the vaccinated animals after 21 days post boosting. The serum thus separated was purified through ammonium sulfate salt (NH4)2S04 and ion exchange column chromatography. Total protein content in the guinea pig serum (whole serum), Ammonium Sulfate Precipitated Guinea Pig Serum (ASPGPS) protein and Ion Exchange based purified Guinea Pig Serum (IEGPS) protein when analyzed through spectrophotometer at 280 nm and 260 nm was found to be 52 ug/mi, 24 ug/ml and 10 ug/mi respectively. Virus Neutralization (VN) test was performed to monitor the neutralizing antibody titer. The whole serum of guinea pigs and rabbits showed the 1:32 and 1:64 anti-FMD serotype "0" virus neutralizing antibody titers. While anti-FMD serotype "0" virus neutralizing antibody titer was 1:128 in the IEGPS proteins. IEGPS protein with 1:128 neutralizing antibody titer were used as capture/trapping antibody in the standardization of the assay. The IEGPS protein 1:1000 diluted with 10 ug/ml of protein content was found to be optimum as capture/trapping antibody. To cover residual blank spaces, different available blocking buffers were evaluated and Skimmed Milk Solution 5 % in Phosphate Buffered Saline (PBSSKIVI-5%) proved best amongst blocking buffers. Coating of 1:1000 diluted IEGPS at 37 °C for 1 hour followed by storage at 4 °C for overnight was best incubation time in the study. FMD serotype "0" virus 1:100 diluted was optimum in IS-ELISA. Similarly rabbit anti-FMD serotype "0" virus specific immune serum 1:10,000 diluted and goat anti-rabbit IgG horseradish peroxidase conjugate 1:4000 diluted were found to be optimum during the standardization of the assay. Lastly ELISA plates were proved to be best amongst the available plates for assay. In each experiment, plateau region, test background and plate background were recorded. Results of the study helped for establishment of an economical, sensitive, reliable, robust IS-ELISA technique in research and diagnostic laboratories in the country. Availability: Items available for loan: UVAS Library [Call number: 0883,T] (1).

34. Plasma Concentration Of Doxycycline After Flock Medication Via Drinking Water In Broilers

by Ayaz Ali Khan | Prof.Dr.Muhammad Ashraf | Dr. Khushi | Dr. Muhammad Ovais Omer.

Material type: book Book; Format: print Publisher: 2004Dissertation note: An experiment was conducted on the bioavailability of doxycycline in broiler chickens after multiple oral administration at the dose rate of 10mg/kg.b.wt. under field condition and comparison was made between the two different pharmaceutical preparations of doxycycline. Stability of doxycycline solution in drinking water was also evaluated in the study. Microbiological assay was used to determine doxycycline concentration in plasma and water samples. Doxycycline was given to 2 groups of 30 chickens. Maximum plasma level of 4.116 0.326 g/ml (group 1) and 4.00 0.280 g/ml (group 2) was obtained after doxycycline administration, having no significant difference. Minimum plasma level of 1.566 0.202 g/ml (group 1) and 1.116 0.116 g/ml (group 2) were observed during the field experiment. Both the formulations attained peak levels of the plasma concentration at the same time which was 6 3.46 hours. Mean SEM of the area under curve (260.933 15.043 g.hr/ml) of group 1 had no significant difference from that (246.383 13.187 g.hr/ml) of group 2. Statistical analysis revealed bio equivalency between the two preparations of doxycycline. The residual doxycycline HCl content of doxycycline HCl solution was 89.72% (group 1) and 81.6% (group 2) after every 4 hours expressed as the percentage of the initial concentration. Availability: Items available for loan: UVAS Library [Call number: 0890,T] (1).

35. Studies On Nonsteroidal Anti-Inflammatory Drugs (Nsaids) Toxicity In Broilers

by Asif Farooq Awan | Prof.Dr.Muhammad Ashraf | Dr. Khushi | Dr. Muhammad Ovais Omer | Faculty of Biosciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: This project was designed for the evaluation of different effects of toxic dosage levels of NSAIDs (piroxicam, ketoprofen, phenyl-butazone and dipyron) in broiler chickens. For this project two hundred and twenty five healthy broiler chickens were purchased from the market and were reared upto 28 days. Then one hundred and twenty five were divided into five groups A, B, C, D and E having twenty five birds in each group. On day 29th four groups A, B, C and D were medicated with piroxicam, ketoprofen, phenyl-butazone and dipyron twice a day at dose rate of 1, 5, 50 and 50 mg/kg body weight respectively intra-muscularly for four days. Birds from group E were kept as control. Feed and water were provided ad libitum. A physical examination was performed daily. Signs of toxicity and mortality rate in each group was recorded. Blood samples from wing vein (3 ml) was drawn on day 29 before medication and on days 33, 37, 41 after medication from same birds for determination of serum values of Aspartate Transaminase (AST), Alanine Transaminase (ALT), Uric Acid, Alkaline phosphatase (ALP), and Creatinine. Postmortem examinationwas performecd after all samples taken. In second experiment other 100 birds were divided into five groups K, L, M, N and O comprising of 20 birds in each group. Each bird of group K was injected I/M piroxicam 2 mg/kg twice a day. Each bird of group L was injected I/M ketoprofen 10 mg/kg twice a day. Each bird of group M was injected I/M phenyl buazone 100 mg/kg twice a day. Each bird of group N was injected I/M metamizole 100 mg/kg body weight for twice a day upto four days and group 0 was kept as control group. Postmortem examination was performed after medication. Based on the necropsy findings and biochemical analysis it was found that piroxicam was safest drug (NSAIDs) in the avian species. Keeping in view the environmental problem of decline in population and its possible link with Diclodenic sodium,(vultures arises) it is recommended that piroxicam which has good pharmacological effects in human medicines may be used instead of diclofenac sodium in veterinary practice. Availability: Items available for loan: UVAS Library [Call number: 0892,T] (1).

36. Studies On In Vitro Culture Characteristics Of Adherent Baby Hamster Kidney-21 (Bhk-21) Cell Line

by Saddeq-ru-Rahman | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2005Dissertation note: Baby Hamster Kidney-2 1 (BHK-2 1) cell line growth pattern, growth requirements, growth effectors, cryopreservation and its susceptibility to different animal viruses were studied to optimize the in vitro culture requirements and conditions for maintenance and long time cryopreservation in liquid nitrogen of this cell line. It was observed that BHK-2l cells multiplied fast during first 48 hours and made a complete monolayer after getting confluency with in 72 hours post incubation which was followed by a decline phase. Fetal calf serum has growth stimulating effect and 5 - 10% serum level was satisfactory for the maintenance of cell line. While harvesting the cells from a flask, Trypsin (0.25%) with neutralization by fetal calf serum (5-10%) was found better. For cell storage 10% Dimethylsulfoxide (DMSO) through gradual cooling maintained maximum recovery of viable cells during cryopreservation. Footh and mouth disease virus (FMDV; serotype "0" and "A") were adapted to cell this cell line, while canine parvo virus (CPV), Newcastle disease virus (NDV LaSota strain), canine distemper virus (CDV), and hydropericardium syndrome virus (HPSV) could not adapted to this cell line through five blind passages in this study. Availability: Items available for loan: UVAS Library [Call number: 0916,T] (1).

37. Preparation And Evaluation Of Cell Culture Vaccines Against Hydropericadium Syndrome Virus In Poultry

by Jamshid Akhter | Masood Rabbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: In this study a total of 9 vaccines were prepared, 6 vaccines were prepared from cell culture passaged HPS virus having TCID50 i' and inactivated with formalin and binary ethyleneimine (BET ) with and with out different adjuvant combinations. While other 2 vaccines were prepared from more diluted virus suspension with PBS (10 and 100 times) and were inactivated by binary amine and adjuvant was oil base. The 9th vaccine was prepared from infected liver homogenate (aqua base) and inactivated with formalin. These vaccines were evaluated in 10 groups of day old broilers (100 chicks) for their comparative immunogenicity and protection. At day 14 of age, groups Al, B 1, Cl, and C were vaccinated with four oil base vaccines with different virus concentration and different inactivants. Similarly groups Dl & D were vaccinated with aluminized vaccines and groups A, B, and El with non adjuvant vaccines. Groups E was kept as unvaccinated control group. Serum samples were collected from all groups on 0, 14, 28 and 42 day of age and subjected to AGPT for seroconversion. AGPT GMT results indicated difference for different virus concentrations and no difference for different virus inactivants but same adjuvant. The AGPT GMT recorded 0 & 14 day of age pre vaccination indicated the maternal antibodies against HPS in chicks were not protective level. The chicks became protective against the disease in most susceptible age. The AGPT GMT recorded 14 and 28 days post vaccination indicated the highest and more consistent (149.2 and 182) for oil base vaccines with virus concentration having TCID50 104.1 but for vaccines of diluted virus suspension then GMT was variable. Similarly aluminized vaccines showed (149 and 94.4) and non adjuvant cell culture vaccines showed (116 and 2.7) while non adjuvant liver homogenate showed (80.5 and 2.3). On day 42 of age, all birds were challenged with virulent HPS virus and percentage mortality and percentage protection for each group were recorded. Lowest mortality (0%) and highest protection (100%) were recorded for groups vaccinated with oil base vaccine. There was zero mortality and 100 percent protection were recorded for group Dl (BET inactivated) while in group D (formalinized) there was 10 percent mortality and 88 percent protection in groups vaccinated with aluminized vaccines. While mortality and protection in groups vaccinated with non adjuvant cell culture vaccines were 25% and 71.5% while in group vaccinated with non adjuvant liver homogenate vaccine was 50% and 44%, respectively. Cell culture oil base vaccine against HPS virus, having io' TCID50 inactivated with BET was concluded the best in experimental trails and has been recommended for commercial production after field evaluation. Availability: Items available for loan: UVAS Library [Call number: 0918,T] (1).

38. Standardization Of An Indirecto Enzyme Linked Immuno Sorbent Assay Elisa For Measuring Antibodies Of Infectious Bursal Disease Virus

by Faisal Amin | Dr. Mansur-ud-Din Ahmad | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2005Dissertation note: This project was conducted to make an attempt to develop an in-house ELISA to measure antibodies against IBDV. ELISA is the most commonly used serological test for evaluation of IBDV antibodies, but the cost of imported ELISA kit is usually very high and not affordable by the average farmer. The present study was designed with an aim to develop a cost effective ELISA kit under local conditions. Various steps involved in the development of ELISA were standardized. Two types of antigens i.e. "A" and "B" were used. Antigen "A" was prepared by propagating the live IBD virus (D-78, Intervet) in CEF cells and further concentrated by dialysis against PEG-6000. Antigen "B" was the reconstituted live IBDV vaccine. Both the antigens produced acceptable and comparable results but antigen "B" is conventional due to ease of preparation and to avoid a time consuming and costly procedure of cell culture. The optimum dilution for antigens "A" and "B" were found to be 1:300 and 1:600 respectively. The optimum dilution of conjugate was selected as 1:2000 and incubation time was standardized as 30 minutes at room temperature (25-30°C) after addition of ABTS as substrate. Standard curve (two fold dilution of the positive sera up to 1 1th dilution) was constructed and 3 standards were selected to cover the range of strong reactor with non-reactor sera. The in-house developed ELISA was evaluated with field chicken sera samples of different age groups. The sera samples of different groups (A, B, C and D of 1-day-old broiler breeder chicks, 1-day-old broiler chicks, 13 weeks old vaccinated layer breeder bird and 30 weeks old vaccinated broiler breeder birds respectively) were evaluated with in house ELISA and its efficiency was compared with commercially available ELISA kit. The samples that were either positive (strong or weak) or negative with commercial ELISA kit also had similar pattern with in-house ELISA. Groups A, C and D were strongly positive while group B was found negative for IBDV antibodies. It is concluded that in-house developed ELISA is comparable with the commercially available kits. Availability: Items available for loan: UVAS Library [Call number: 0930,T] (1).

39. Studies On The Physico-Chemical Factors Affecting In Vitro Replication Of Foot And Mouth Disease Virus (Serotype"O")

by Muhammad Taslim Ghori | Prof. Dr. Khushi Muhamamd | Prof. Dr. Masood Rabbani ( | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2007Dissertation note: Effect of physical (temperature, U.V light, and pH) and chemical factors such as sodium carbonate, sodium hydroxide, potassium permanganate, formaldehyde, citric acid, fin-virus, iodine and sodium hypochlorite on the replicating ability of"O" type of FMD virus on BHK cell line was determined. The FMD virus of known TCID was exposed to 37, 57 and 77°C for 15, 30 and 45 minutes. Each of the virus's aliquot exposed to temperature was inoculated to 8 wells of the 96-well cell culture plate containing adherent monolayer of BI-IK cell line. The plates were incubated at 37 °C for 48 hours. The results showed that the temperature more than 57°C can inactivate the virus within 15 minutes. The virus was admixed in the MEM-199 maintenance media at pH 3, 5, 7, 9 and 11. The results showed that the virus was survived at pH 7 but virus was inactivated at pH 3, 5, 9 and 11. The FMD virus of the known TCID o was exposed to U.V. light (1 foot distance) for 15, 30 and 45 minutes. The results indicated that the virus tolerated UV light of 252.7nrn as it showed cytopathogenic effects (CPE). The FMD virus of known TCID was exposed to 0.5 x, lx, and 2x concentration of each of the iodine, sodium carbonate, sodium hydroxide, formalin, finvirus, potassium permanganate, sodium hypochlorite and citric acid for 30, 60 and 90 minutes. Each of the virus's aliquot exposed to either of chemical factors was inoculated to 8 well of the 96-well culture plate containing adherent monolayer of the BI-IK cell line. The plates were incubated at 37 °C for 48 hours. Development of CPE indicated the virus inactivating ability of the chemical factor. The results showed that formalin, iodine, finvirus and sodium hypochiorite are more effective against FMD virus. The results of the study are helpful to curtail the spread of virus, to implement the effective bio-security measures in our local conditions and in processing of animal products fit for export. Availability: Items available for loan: UVAS Library [Call number: 0955,T] (1).

40. 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 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).

41. Effects Of Different Disinfectatnts On Pathogens In Poultry

by Asif Abbas Malik | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2006Dissertation note: Poultry sector is the second largest industry after textile in Pakistan. It is threatened by various diseases i.e; Newcastle disease (ND), Avian Influenza (Al), Colibacillosis, Hydro-pericardium syndrome (HPS) and Infectious bursal disease (IBD, Gumboro). The efficacy of various available disinfectants (Hygen 275 — 2000 H, Virkon S and Aldekol) was tested at 2x, lx and Y2 x dilution against Staphylococcus aureus, Escheria Coil, Newcastle Disease virus and Avian Influenza virus. Each dilution of all the disinfectant was divided into 4 aliquots i.e; a, b, c and d (each of the aliquot, for each pathogen). Each aliquot were mixed with equal volume of either of the pathogen. The mixture of the disinfectant and the pathogen was incubated at 37°C for a period of 15, 30 and 45 minutes of interaction. The contents were collected aseptically and processed to evaluate the effectiveness of the disinfectants. Disinfectant A (Hygen 275-2000H) showed good bactericidal as well as virucidal activity at 1% dilution. Disinfectant B (Virkon S) was able to kill all the bacteria and viruses even at 0.25 % dilution. While, disinfectant C (Aldekol) effectively killed the bacteria and viruses at 0.5 % and I % dilutions. Results of the study will help the farmers to adopt effective biosecurity measures to minimize the challenges at farm level. Availability: Items available for loan: UVAS Library [Call number: 0966,T] (1).

42. Isolation And Characterization Of Clostridium Perfringens From Domestic Animals An Man In Punjab

by Waheeda Raana | Prof. Dr. Muhammad Akram Muneer | Dr. Khushi Muhammad | Prof. Dr | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2007Dissertation note: The objectives of present investigation were to isolate the Cl. perfringens from the domestic and zoo animals and human beings; characterize it through biotyping and pathogencity observation, and to develop a vaccine- from the common CI. perfringens isolate. For this purpose a total of 1240 samples of morbid tissues (faecal samples from animals and gangrenous tissues from humans). From cattle (n=180), goats (n=180), horses (n=250), camel (n=250), deer (n=28), wild beast (n=07), monkeys (n16), zebra (n10), elephant (n01), yaks (n=09), foxes (n07), jackals (n=08), baboons (n=08), and bears (n08) were collected and processed for isolation of CI. perfringens. In addition a total of 100 human cases; 83 wound swabs and 17 gas gangrene were also collected and analyzed bacteriologically. This study has indicated that Clostridium (Cl.) perfringens causes multiple clinical problems in animals and human beings as was indicated by good rate of its isolation from the examined morbid tissues and fecal samples. Of the total 1240 samples from various types of animals 297 (23.95%) indicated the presence of CI. perfringens. The overall isolation percentages of various types of CI. perfringens from the cattle, sheep goat, horses, camel, wild beast, deer, bear, jackal, zebra, monkeys, yak, elephant, baboon, foxes, and humans were 22.2, 12.2, 57.2, 8.0, 21.6, 57.1, 30.76, 37.50, 50.0, 50.0, 37.50, 33.33, 100.00 75.00, 57.14 and 18.00, respectively. Of the tested population of domestic animals, goats indicated the highest Cl. perfringens (57.2%) infection rate. In the zoo animal population, the elephant, baboons, wild beast, jackals, and foxes were shown to be heavily infected with various CI. perfringens types compared to other wild life animals species. Of the 298 isolates obtained through this investigation Cl. perfringens type D was obtained from 118 (39.7%) morbid samples of the domestic and zoo animals; CI. perfringens type A from 63 (21 .21%) samples, Cl. perfringens type B from 95 (31.98%) samples; and the CI. perfringens type E was isolated from 21(7.07%) samples. None of the samples indicated the presence of CI. perfringens type C. Of the total 100 samples from the humans, CI. perfringens type A was isolated from 14 (14%) and Cl. perfringens type D was isolated from 04 (4%). None of the human samples showed the presence of Cl. perfringens types B, C, or E. Of the 17 human gangrene tissue samples, Cl. perfringens type A was isolated from 09 (52.94%) samples and the Cl. perfringens type D was recovered from 02 (11 .76%) samples. However, all attempts to isolate Cl. perfringens types B, C or E from the human gangrene tissue/material samples were unsuccessful. The overall findings indicated that of the total 297 samples positive for various Cl. perfringens types 63 indicated the presence of Cl. perfringens type A. Of those 63 Cl. perfringens type A isolates, 49 were recovered from the animals; and 14 were isolated from the wound swabs and gangrene tissue material samples from humans. Of the 63 Cl. perfringens type A isolates from the animals, 5 were isolated from cattle; 3 from sheep, 20 from goats; 5 from the horses; 10 from camels, 01 from the deer; 01 from the zebra, 01 from baboon, 01 from fox, 01 from the monkey, and 01 isolate was recovered from yak. Of the 14 isolates of Cl. perfringens type A from humans, 05 were recovered from the open wound swabs, and 09 strains of the organism were isolated from the gangrenous tissue material. Of the 297 samples positive for various Cl. Perfringens types, 95 animal samples indicated the presence of Cl. perfringens type B. These 95 isolates were obtained from cattle (n=22), sheep (n=10), goats (n=30), horses (n=03), camel (n=14), deer (n03), wild beast (n=02), monkey (n=02), zebra (n=02), yak (n=01), fox (n01), jackals (n02), baboon (n02) and bear (n=02). None of the human samples was positive for Cl. perfringens type B. Isolation of C/. perfringens type B from the zoo animals is a matter of concern for the human health, as the zoo visitors have the possibility to get infected with this organism. Of the total 297 positive samples of faecal and morbid tissues from various types of animals and human being Cl. perfringens type D isolates were recovered from 118 (39.7%) samples. Of these 118 isolates of Cl. perfringens typeD, 114 were obtained from various types of animals, and 04 isolates were from the humans. Of the 114 animal isolates, 10 from the cattle, 5 from the sheep, 44 from the goats, 9 from the horses, 27 from the camel, 4 from the deer, 02 from the wild beast, 02 from the monkey, 02 from the zebra, 01 from the elephant, 01 from the yak, 02 from the fox, 02 from the jackals, 02 from the baboon, and 01 isolate the bear. A total of 04 CI. perfringens type D isolates were recovered from gangrenous tissue and open wound samples from human beings. During this investigation 21 isolates of CI. perfringens Type E were obtained from domestic and zoo animals. Of the 21 isolates, 03 were from cattle, 04 from sheep, 09 from goats and 03 from horses, 01 from monkey, and 01 from the baboon. All the 21 isolations were from the fecal material of above mentioned animals. None of the human samples was positive for CI. perfringens type E. Alpha toxin was produced by all of the 63 Cl. perfringens type A isolates. Within the toxin producing isolates, there was no difference in the quality of toxin in respect to its lethality for mice, dermonecrosis effects for guinea pigs and cytotoxicity in the HeLa cells. The 07 fecal isolates were hemolytic, lecithinase (+), and positive for all biochemical characteristics of Cl. perfringens. Those isolates were not lethal for mice, indicated no dermonecrotic activity in guinea pig, and produced mild degree of cytotoxicity in the cell cultures. The activity of beta toxin obtained from 95 isolates of CI. perfringens type B isolates was determined using standard toxin-antitoxin test carried in mice and the standard serum neutralization test with antitoxin raised in rabbits. Within the toxin producing isolates, no difference was seen in the potential of toxin based on its lethality for mice. Epsilon () toxin activity of the 114 isolates of CI. perfringens type D from animals and 4 of the human isolates was also determined. Of the 114 animal isolate, 110(96.49%), and all the 4 human isolates produced E-toxin. There was no difference in the lethal potential of toxin for mice, dermonecrotis action in guinea pig and production of CPE in VERO cells. Iota (i) toxin activity of the 21 isolates of Cl. perfringens type E was also determined serum neutralization test in mice. Many isolates produced more than one major toxin. Ci. perfringens (CP) type A produced Alpha (a) toxin; CP type B produced Alpha (a), Beta (3) and Epsilon (E) toxins; OP type D isolates produced Alpha (a) and Epsilon (E) toxins, and OP type E isolates produced Alpha (a) toxin + Iota (i) toxin. The immunobiologic studies of isolates showed that many of the isolates were quite antigenic. Isolates of CI. perfringèns type D and B were found highly immunogenic as those isolates producing SN titer of 1:320. Availability: Items available for loan: UVAS Library [Call number: 0998,T] (1).

43. Dvelpoment And Optimization Of Multiplex Pcr For The Detection Of Avian Influenza Strains In Pakistan

by Mirza Salman Saleem | Asso. Prof. Dr. Muhammad Hanif | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2009Dissertation note: The pathogenic Influenza A viruses (subtype H5N1, H7N2 and H9N3), are emerging avian influenza (AI) viruses that have been causing global concern as a potential pandemic threat. Some forms having zoonotic importance (H5N1 and H7N7). So it is a matter of priority to develop quick and efficient methods for detection of Influenza viruses. For the detection of avian influenza, HA (haemagglutination) test and HI (haemagglutination inhibition) tests are being used for long time. But studies have shown that Influenza virus shows variability and diversity and a high rate of mutation, which makes diagnosis difficult. For this reason the reverse transcriptase PCR (RT-PCR) assays are considered to be a helpful tool. In this study design, a multiplex RT-PCR strategy was optimized and developed for the detection of AI virus (subtypes H5, H7 and H9). Primers were designed from sequence available Influenza Database (IVDB) for Pakistan and neighboring regions. The primers were annealed at different temperatures so as to optimize a temperature at which all three primers can amplify their respective subtypes. The results clearly indicated that a multiplex RT-PCR is a quick and efficient method for the detection and it is also economical as fewer reagents are utilized. The PCR products of the reaction can potentially be used to provide additional information about strain variation, either by restriction analysis or PCR product sequencing. The core objectives achieved are the development of an efficient and economical method for detection of avian influenza viruses by designing indigenous primers and optimization of a multiplex RT-PCR for the avian influenza virus. Availability: Items available for loan: UVAS Library [Call number: 1148,T] (1).

44. Identification And Genotyping Of Vp1 Genses Of Fmd Viruses

by Atia Bukhari | Prof. Dr. Irshad Hussain | Prof. Dr. Khushi Muhammad.

Material type: book 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. Availability: Items available for loan: UVAS Library [Call number: 1179,T] (1).

45. Effect Of Differnet Physico Chemical Substances On The Production Peotential Of Phycocyanin From Spirulina and its Characterization

by Firasat Hussain | Dr. Imran Najeeb | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Spirulina is a multi-cellular, filamentous Cyanobacterium, belonging to a blue-green alga of Cyanophyta. Spirulina is recently proven in animal experiments to exhibit various biological activities such as lowering plasma cholesterol levels and blood pressure. The principal phycobiliproteins present in spirulina are phycocyanin and allophycocyanin which are made up of dissimilar ? and ? polypeptide sub units. The fresh biomass was found suitable for phycocyanin extraction. Freezing and thawing of cells was proved the best method for extraction of phycocyanin (0.4mg/ml), as compared to homogenization, hydrochloric acid and sonication. Nitrogen effects phycocyanin production from spirulina. Different concentrations of nitrogen spirulina medium were provided. Among which 1.875g/L spirulina produced phycocyanin (0.412mg/ml). Phosphate effects phycocyanin production from spirulina. Different concentrations of phosphate spirulina medium were provided.Among which 1.5g/L spirulina produced phycocyanin (0.354mg/ml). There is also effect of temperature on phycocyanin production. Spirulina medium 0.192mg/ml at 25oC, 0.390mg/ml at 30oC, 0.184mg/ml at 35oC. There is also effect of light on phycocyanin production. 0.361mg/ml were produce at 1500 Lux. Molecular weight (66kDa) of phycocyanin was confirmed by SDS-PAGE and explored potential production of phycocyanin from indigenous spirulina. Availability: Items available for loan: UVAS Library [Call number: 1204,T] (1).

46. Characterization Of Indigenious Species Of Mycotoxins Producing Aspergilli

by Gull Naz | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Pakistan's economy is based on agriculture. Agriculture crops are harvested and stored in feed mills for production of thousands ton of feed for livestock as well as poultry through out the year. In Pakistan, July and August are hot and humid months during which moulds grow abundantly on the heaves of wheat! rice/maize straw and feed ingredients and produce variety of toxins. Present study has been designed to explore different groups of moulds prevailing in and around Lahore city in each month of the year. Samples of soil and air were collected from ten different places of Lahore city. A total of 240 samples were cultured on a common Saboraud's Dextrose Agar to get single colonies of each mould. These single colonies were identified by colony characters, slide cultures and biochemical tests. Mycotoxin producing Aspergilli were isolated by culturing on specified media and placing the cultures under Wood's lamp. Mycotoxin productions potential were assessed by extracting mycotoxins of these Aspergilli. Mycotoxins produced by the Aspergilli were identified and purified through Thin Layer Chromatography. These mycotoxins were then quantified through High Performance Liquid Chromatography. The identified and purified mycotoxins can be used as standards. Reference standards are important and critical for qualitative and quantitative detection of mycotoxins in field samples screening. Presently mycotoxin is a ban item. The occurrence of toxinogenic Aspergilli have economic impact directly on livestock and poultry products export. Availability: Items available for loan: UVAS Library [Call number: 1217,T] (1).

47. Effectof Mentofin (Herbal Product)On Antibody Response Of Broilers To Newcastle Disease Vaccine

by Saif- Ur- Rehman | Prof. Dr. Khushi Muhammad | Dr. Tahir Yaqub | Prof. Dr. Muhammad.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Immunostimulants such as Mentofin® are commonly used to enhance the immune response of birds to vaccines. In the present study immunomodulatory effect of Mentofin® on antibody response of broilers to Newcastle Disease virus vaccine was evaluated. For this purpose one hundred one day old broilers were divided in to four groups (A, B, C and D) each containing 25 birds. Each bird of group A and B was vaccinated against ND and each bird of group A and C was treated with Mentofin. Anti-ND-HI antibody titer of each bird of each group was monitored on 14, 21, 28 and 35 days of age. Mentofin® treated broilers showed higher consistent antibody (anti-NDV HI antibody titer) response as compared to untreated broilers. These birds when given challenge infection of velogenic ND virus on 35 days of age showed same protection as that of untreated vaccinated birds. However non vaccinated broilers treated with Mentofin showed higher protection as compared to that of in non treated unvaccinated birds. Weight gain in Mentofin® treated broilers was same as that of non-treated birds. Similarly, there was no effect of the Mentofin on FCR. Droppings from Mentofin treated birds showed no urease producing bacteria while 100% droppings of the herbal untreated birds showed urease producing bacteria. Mentofin at 1% concentration in nutrient broth inactivated the proteus species while its 0.0001 % concentration inactivated the bacteria in urea broth. In in vitro studies, 0.5 % concentration of Mentofin inactivated the lentogenic strain of ND Virus within 15 minutes at interaction temperature of 37 0C. The results can be used to formulate the vaccination policy along with herbal based immunostimulatant such as Mentofin®. Availability: Items available for loan: UVAS Library [Call number: 1294,T] (1).

48. Passive Immunization Of Infectious Bursal Disease Virus Infected Birds Using Chemically Purified Immune Yolk

by Ammara Akram | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2011Dissertation note: Infectious bursal disease (IBD) is a major killer disease of poultry. It is also known as Gumboro disease where the disease was reported first time. It is double stranded RNA virus belongs to the family birna viridae. This disease is quite endemic in Pakistan which has huge impact on poultry industry. Besides vaccination if immune yolk is properly harvested and purified it can be used for treating of IBDV infected birds. Therefore this work has been outlined to study the effectiveness of immune yolk in experimentally produced IBDV infected birds. Refinement of yolk IgY from egg yolk of immunized hens. Suitability in using hyperimmune egg yolk in IBD infected bird in field conditions. In order to get hyper immunized egg yolk 20 commercial layers were raised in poultry shed of the university. They were supplied with fresh water and feed ad libitum with proper hygienic condition. The birds were vaccinated with oil based killed Gumboro vaccine twice at 15 days interval at the age of 26 weeks to get immune yolk. Eggs were collected at two weeks interval till two and half months after boosting. Immune yolk was purified by chemical means. Antibody against IBD in egg yolk and semi purified egg yolk IgY was measured by indirect ELISA kit method. The eggs which were collected 15 days interval after boosting had the highest antibody titre which decline with the passage oftime and lowest was recorded 75 days after boosting. Similar pattern of results were also observed in semi purified egg yolk. However significant antibody titre was lost during purification process. 50 commercial chicks of 15 days old were purchased and they were reared in poultry shed in the university up to 36 days. They were splitted in eight groups and two experiments were carried out side by side. In experimental chicks the birds were challenged with the Gumboro infected bursal homogenates which were confirmed by agar gel diffusion tests. In first experiment the birds were challenged at the day of 30 days and they were provided with passive therapy of immune yolk and semi purified IgY after 3 days of challenge. In the second experiment the birds were challenged and passive immuno therapy was provided 24 hours interval of challenge and concurrently. The birds which received semi purified immune yolk and antibody titre having more or less 4000 they showed 20% mortality in the each group. Availability: Items available for loan: UVAS Library [Call number: 1355,T] (1).

49. In Process Quality Control Factors Affecting Sensitivity Of Rapid Serum Agglutination Antigen Of Mycoplasma

by Rana Khurram Khalid | Prof. Dro. Masood Rabbani | Prof. Dr, Khushi Muhammad.

Material type: book Book; Format: print Publisher: 2012Dissertation note: Mycoplasma gallisepticum (MG) is one of the smallest self replicating infectious agent, it lacks cell wall so cell membrane is the outer most boundary. Its cell membrane is made up of sterols which not only gave it rigidity but also make it fastidious to grow. MG is the causative agent for chronic respiratory disease (CRD). Isolation identification and serodiagnosis are routinely used in laboratories. However among the serodiagnosis rapid serum agglutination (RSA) test is mainly used for early detection for its sensitivity, rapidity and cost effectiveness. Keeping in VIew the importance of RSA antigen test a study was conducted to prepare standardize RSA antigen from local isolate and compare its sensitivity with a commercial RSA antigen. Molecular characterized local isolate of MG procured from University Diagnostic Lab University of Veterinary And Animal Sciences, Lahore, was grown in 11 different media formulations to identify a media with better antigenic yield. Affects of different in process quality control parameters; bacterial concentration (0.75%, 1 %, 1.25% PCV), diluents (normal saline, PBS, HBSS) inactivants (heat, formalin) and preservatives (thiomersal sodium, sodium azide, phenol) were studied in terms of their influence on the sensitivity of prepared RSA antigens. These prepared antigens from local isolate were further compared with a commercial RSA antigen. The results of antigenic yield were statistically analyzed through One Way A OVA test. All the media formulations support the growth of MG isolate except Frey's media with 7.5% fetal bovine serum and 7.5% chicken serum that revealed no growth or packed cell volume. The maximum bacterial growth in terms of packed cell volume was obtained from frey's media with 12% horse serum. So this media was further use in the production of antigens. Sensitivity of RSA antigens with different bacterial concentrations in terms of packed cell volume, preservatives, inactivants was more using HBSS followed by PBS and normal saline. However difference in increasing the sensitivity of RSA antigen in terms of agglutination with known serum was statistically found significant. Formalin inactivated RSA antigens were somewhat more sensitive as compared to the heat inactivated using different diluents and bacterial concentrations. Sensitivity of RSA antigens in different diluents, inactivants and preservatives was a little bit more with 1.25% pev followed by 1.00% pev however it was least with 0.75% Thiomersal sodium added RSA antigens were more sensitive followed by phenol and sodium azide. PBS and HBSS based, formalin inactivated and preservative (thiomersal sodium, phenol, sodium azide) added antigens having bacterial concentration of 1.25% and 1 % (peV) given agglutination at 1 :30 dilution of known positive MG serum. The sensitivity of all these antigens was equal to the commercial RSA antigen. All the prepared RSA antigens and commercial anitigen showed no agglutination with known negative serum. Sensitivity of most of the prepared and a commercial antigen was comparable. The findings of this study will be helpful for further recommendations of local RSA MG antigen used in the diagnosis of chronic respiratory disease. Availability: Items available for loan: UVAS Library [Call number: 1406,T] (1).

50. Comparison Of Diagnostic Approaches For The Detection Of Bovine Viral Diarrhea Persistency In Dairy Herds

by Arfan Ahmad | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print Publisher: 2012Dissertation note: Bovine viral diarrhea is one of the most important diseases of cattle which are causing continuous economic losses to the cattle industry primarily due to decreased reproductive I performance. Without doubt, direct contact between BVDV persistently infected, and susceptible animals is the most important transmission route of virus. All control programs which are in use in many countries of the world, mainly depend upon the detection of PI animals, eliminating them and preventing their return into the herds. Therefore, in this study diagnostic suitability of ear notch biopsies and serum samples were compared for the detection of PI animals, as well as proficiency of various diagnostic approaches like VI, AC-ELISA, IHC and real time RT-PCR were evaluated using ear notch biopsies. A total of 468 samples were collected from 12 participating dairy cattle farms located at Prince Edward Island, Canada. The samples were divided into two groups on the basis of age, A " 6 months), and B (> 6 months). PI calves remain immunotolerant to the infecting strain but if exposed to a heterogonous strain postnatally, they may develop low level of antibody. Accordingly, serum neutralization was applied for initial screening of samples for further testing. The samples of animals of group B, having SNT (:S 1 :64) were selected, while all samples of younger aged group A were processed without considering the serum neutralizing titres, because unlike older animals, P.1. animals below 6 months of age can have passive colostral antibodies in the course of persistency. Diagnostic suitability of ear notch biopsy and serum sample for confirmation of BVDV A significant discrepancy was observed between ear notch biopsies (51198 positive) and serum samples (71198 positive) during first round of testing by real time RT-PCR. However, on follow up testing, 30 days post first round of testing, a complete agreement between ear notch biopsies and serum samples was observed. On second round of testing, a total of 4 animals out of 197 (one positive animals died before re-sampling) were confirmed with PI, using both ear notch biopsies and serum samples. The decrease in the positivity using RT-PCR on serum samples in the second round of testing reflected the presence of 2 transiently infected animals. Ear notch biopsy (EN) testing did not detect any transiently infected animal indicating the lack of delectability of the virus in EN during transient infection under conditions of this study. After follow up testing, 2 animals in each of group A and B were identified as PI. These findings have led us to conclude, that either serum or ear notch biopsy can be used for the detection of persistent infection. Of 468 collected and 197 tested samples, an overall 0.85% and 2.03% prevalence of PI animals with BVDV was observed respectively. A complete agreement (P value=l) was observed when three diagnostic approaches (Real time RT- PCR, AC-ELISA, and IHC) were compared with standard of VI. A total of 197 ear notch biopsies (145 of group A and 52 of group B) were tested by the four diagnostic tests, four animals (2 from group A and 2 from group B) were found positive by all the tests applied. A complete agreement was observed between the first and the second round of testing. All four assays were found specific but real time RT-PCR was found to be more sensitive. Both, VI and IHC were found labour intensive, as diagnosis may take more than one week to be made. Further PI calves remain immunotolerant tothe infecting strain but if exposed to a heterogonous strain postnatally, they may develop low leved ofantibody. Accordingly, serum neutralization was applied for initial screening of samples for further testing. The samples of animals of group B, having SNT (:S 1 :64) were selected, while all samples of younger aged group A were processed without considering the serum neutralizing titres, because unlike older animals, P.1. animals below 6 months of age can have passive colostral antibodies in the course of persistency. Diagnostic suitability of ear notch biopsy and serum sample for confirmation of BVDV persistent animals were evaluated by real time RT-PCR. TaqMan probes and primers specific for BVDVI and BVDV2 were used. They were found specific and able to detect 10·s and 10-4 TCID50 units ofBVDVI and BVDV2, respectively. Availability: Items available for loan: UVAS Library [Call number: 1407,T] (1).



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