201.
Comparative Efficacy Of Five Diffrent Brands Of Commercial Newcastle Disease Lasota Viurs Vaccines In Broilers
by Tariq Abbas | Prof. Dr. Akram Muneer | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2005Dissertation note: The aim of the study was to compare five major commercial NDV (LaSota strain) vaccines being used in Pakistan with respect to potency, efficacy, thcrnioslability and influence on production performance in briolier chicks. The representative vials of the live NDV LaSota strain vaccines namely A, B, C, D and E were procured from local market. The vaccines were assayed for 50% infectivity (BID50) and Haemagglutinative ability (HA). A 3-log10 difference oF EJD50 and two- to-eight fold difference of HA activity was found was found among the various vaccines. Onc hundred and fifty day- old broiler chicks were divided into six groups and managed separately. The birds in group I, II, Ill, IV and V were actively immunized against ND on day 7 and 21 using vaccines A, B, C, D and E respectively. The birds in group VI served as unvaccinated control. The serum 1-Il antibody response of the different vaccines was determined 7, 14, 21, 28 days post-vaccination. The birds (n= 15) in all tile groups including unvaccinated control was challenged at day 35 with local virulent field isolate. The HI serum antibody profile and post-challenge mortality pattern revealed a dose- response relation between the virus content, humor-al antibody response and clinical protection. To compare the heat stability, the vaccines were incubated at 4C, 25CC and 40C for a period of 24 hours. There was no remarkable reduction in I IA liter, however slight dips (less than 2 logarithmic units) in LID50 values were found in all the vaccines. All the vaccines caused siginifcant suppression in weight gain leading to a poor performance in terms of Feed Conversion Ration (FCR) and European Efficiency Factor (EEF)
Availability: Items available for loan: UVAS Library [Call number: 0909,T] (1).
202.
An Epidemiological Study Of Nosocomial Infections At Mayo Hospital, Lahore
by Tayyaba Ijaz (Phd) | Prof. Dr. Muhammad Akram Muneer | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2005Dissertation note: The present study was designed to investigate the Prevalence of Etiological Agents of Nosocomial Infections in Mayo Hospital, Lahore-Pakistan of the 32,620 patients studied during 1997-2001; a total of 4502 (13.80%) patients acquired various types of nosocomial infections during their stay at Hospital. Clinical samples collected from various types of patients consisted of 1040 samples of Pus & Wound Swabs, 109 samples of blood; 115 of Pleural Fluids, 286 of Ascetic Fluids, 37 of Cerebrospinal Fluid, 1398 of Urine, 988 of Sputum; 329 of Burn Swabs, 99 of Patient Body Devices and 101 of Fecal and Drainage Material. The routine techniques for isolation. Identification through Biochemical, Serological and Antibiotic Sensitivity Testing were used for studying the Bacteriology of the selected samples. The present findings revealed that from a total of 4502 samples, 1287 Strains of Staphylococci, 429 Strains of Streptococci, 328 Strains of Enterococci, 781 Strains of Pseudomonas, 349 Strains of Enterobacter, 41 Strains of Acinetobacter, 26 Strains of Klebsiella, 140 Strains of Proteus, 1031 Strains of Escherichia, 67 Strains of Serratia, 93 Strains of Haemophilus, 119 Strains of other types of Gram Positive Bacteria, 13 Strains of other types of Gram Negative Bacteria, and 189 Strains of Yeast and Fungi were found as Etiological Agent for Nosocomial Infections.
Availability: Items available for loan: UVAS Library [Call number: 0912,T] (1).
203.
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: ; 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).
204.
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.
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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).
205.
Studies On Antigenic Homogeneity Of Fowl Adenoviruses Causing Hydropericardium Syndrome In Broilers
by Muhammad Tariq Iqbal | Dr. Mansur-ud-Din Ahmad | Dr. Irshad Hussain | Dr. M. Sarwar | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2006Dissertation note: Babesiosis is a highly important disease in the world, caused by the intraerythrocytic protozoan parasites of the genus Babesia. A wide range of domestic and wild animals and occasionally man are affected by this disease, which is transmitted by ticks and has a worldwide epidemiological distribution. While the major economic impact of babesiosis is on the cattle industry, infections also occurs in other domestic animals , including horses, sheep, goats, pigs and dogs.
The present study targeted the carrier cattle infected with Babesia bigemina and Babesia bovis, as they are difficult to detect because of the low numbers of parasites that occur in peripheral blood. However, diagnosis of low-level infections with the parasite is important for studying the transmission and standardizing epidemiological studies. Using the Polymerase Chain Reaction (PCR) to amplify a portion of the gene from the parasite, and tested the ability of this method to detect carrier cattle.
A study was conducted to detect the. Babesia in blood samples through PCR based techniques. A PCR assay was described which could differentiate Babesia bigemina and Babesia bovis by using specific primer in carrier cattle. Blood samples of 100 cattle were randomly analyzed with PCR assay 29 (29.0%) out of 100 blood samples were positive for babesiosis in which 18% were positive for Babesia bigemina and 11% were positive for Babesia bovis, While the Light Microscopy detected only 18 (18%) out of the same samples. The samples found positive by LM were reconfirmed during the PCR assay but no sample was found to be having both Babesia bigemina and Babesia bovis infections simultaneously.
Thus it is concluded that PCR is a reliable molecular diagnostic technique to detect low level of infections in carrier animals in a population and thus could be used as an effective screening tool for the control and eradication of disease.
Availability: Items available for loan: UVAS Library [Call number: 0928,T] (1).
206.
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: ; Format:
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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).
207.
Isolation And Biological Characterization Of H7 Type Avian Influenza Virus
by Syed Abid Hussain | Dr. Muhammad Akram Muneer | Dr.Azhar | Dr.Masood Rabbani | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2006Dissertation note: The present study was undertaken with the objectives to isolate and characterize avian influenza virus responsible for high morbidity and mortality in layers flocks in Karachi area of Pakistan. AIV H7 type was isolated from the morbid tissue samples by inoculating their tissue homogenate in the 9-1 1 day old developing chicken embryos. This isolate later declared as AIV H7 type, induced death of chicken embryos within 36 to 48 hours post inoculation. The type and subtype of the isolate was confirmed using known nionospecific antisera against various haemagglutinating viruses. Infectivity potential of AIV was determined by inoculating it in 9-11 day old embryos of layer, duck, desi-bird and pigeons. The AF from experimentally infected embryos, haernagglutinated the chicken red blood cells. However, the haemagglutination titer of virus in AF from eggs of various avian species was variable. For evaluating the survival/resistance of the virus against various chemical disinfectants, it was inoculated in embryonated chicken eggs following treatment of various dilutions disinfectants like Beloran, Zeptin and Formalin. These disinfectants were effective in inactivating the avian influenza virus at various concentration levels. For monitoring pathogenic characteristics, isolated virus was inoculated in the developing chicken embryos. The mean death time was evaluated as 42 hrs post inoculation. This investigation indicated that AIV-H7 type was widely circulating iii poultry flocks, and was causing high morbidity and mortality in the susceptible populations. The use of disinfectants like Beloran, Zeptin and F'orrnalin is suggested for inactivating the virus present in-and-around poultry farm premises.
Availability: Items available for loan: UVAS Library [Call number: 0937,T] (1).
208.
Comparative Study On The Pathogenicity Of Liver Homogenate And Cell Culture Propagated Hydropericardium Syndrome
by Shahid Zaman | Dr.Mansur-ud -Din Ahmad | Prof. Dr.Muhammad Akram Muneer | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2006Dissertation note: Abstract Availability: No items available
209.
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: ; Format:
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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).
210.
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: ; 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).
211.
Immmunobiolotical Observations On Avian Influenza Virus Types H7 And H,
by Shahid Iqbal | Prof. Dr. Muhammad Akram Muneer | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2007Dissertation note: The present study was designed to 'find the prevalence of Avian Influenza disease in and around Lahore in commercial and household poultry. A total of 1000 blood and 500 cloacal swabs were collected from Broilers, Broiler-Breeders, Ducks, Pigeons, Sparrows, Quails and Desi Chickens. The blood samples from all the flocks showed non-significant titers while vaccinated flocks showed protective titers. All the cloacal swabs were negative for virus isolation.
The final conclusions from this study were the following.i.e.
- Avian influenza caused by H7 & H9 type is not prevalent in broiler and broiler breeders in and around Lahore.
- The vaccinated poultry flocks showed higher titers of antibodies as compared to non-vaccinated flocks which means that vaccine can play a vital role in protection of bird from H7 & H9.
Availability: Items available for loan: UVAS Library [Call number: 0963,T] (1).
212.
Seroprevalence Of Bovine Brucellosis In District Quetta, Balochistan
by Muhammad Shafee | Prof. Dr. Masood Rabbani | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2007Dissertation note: The sero-epidemiological study of bovine brucellosis was carried out to observe the incidence of brucellosis in slaughterhouse and Government and private dairy farm, (GDF, PDF) Quetta, Balochistan. The prevalence of this disease out of 780 serum samples of cattle and buffalo in slaughterhouse was recorded 3% by Rose Bengal Plate Test (RBPT) and 3.20% by indirect enzyme linked immunosorbent assay (i-ELISA), respectively. The zoonotic natures of this disease was also checked by screening 20 serum samples of slaughterhouse workers butchers and veterinarians and were found (5%) 01 positive out of 20 by RBPT but no positive case was found by i-ELISA. Similarly the disease was also checked in 200 milk samples of Government and Privatly owned Dairy Farm, Quetta.
The overall prevalence observed in the area by screening 1000 serum and milk samples of the target human, cattle and buffalo, was 4.2 % through i-ELISA.
The prevalence observed in Government Dairy Farm (GDF), Quetta was 14.8% (11 positive out of 74) while the Private Dairy Farm (PDF), exhibited 4.76% (6 positive out of 126 milk samples) by screening through i-ELISA. At GDF, Quetta, out of total of 74 cattle, no case were found positive by MRT, although 03 cases were found doubtful while i-ELISA show 11 positive cases in cattle (14.8%) while in private dairy farm 4 out of 15 cattle (26%) were found positive and 01 was considered doubtful by MRT and ELISA detected 06 cases of cattle out of 15(40%). Similarly 2 out of Ill (1.8%) buffalo were positive and 02 were doubtful by MRT but ELJSA did not detect any positive case and the prevalence of bovine brucellosis was higher in animals with reproductive disorders especially in cases of abortion.
The present study also revealed that the disease is more prevalent in cattle than buffalo both in slaughterhouse and organized dairy farm (Govt and private). In slaughterhouse 12 out of 23 cases were found positive by RBPT and 22 out of 23 were found positive by i-ELISA while in organized dairy farm all of the 17 milk samples were found positive from cattle population.
The efficacy of the i-ELISA both for milk and serum samples was found higher than other two conventional tests (MRT and RBPT), as it detected higher percentage of brucellosis cases both in serum and milk samples in comparison to other two tests.
The results of this study have revealed an alarming situation of bovine brucellosis in our dairy animals, which needs an emergent response from policy makers, as the disease is a potential threat to the human and animal health.
Availability: Items available for loan: UVAS Library [Call number: 0964,T] (1).
213.
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: ; Format:
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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).
214.
Pcr-Based Diagnosis Of Canine Parvovirus In Dogs
by Farhan Towakal | Prof.Dr.Masoos Rabbani | Prof.Dr.Khushi Muhammad | Prof.Dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2007Dissertation note: Canine parvovirosis, caused by a haemagglutinating canine parvovirus (CPV), one of the most important acute viral infectious diseases of pups, had been prevalent in the country. In the present study, 40 faecal samples, from clinically suspected cases of parvovirus diseased dogs and stray dogs were collected. The samples were diluted and centrifuged to collect the supernatant. Being a haemagglutinating (HA) virus; the pre-filtered supernatant from all suspected samples was checked for any HA activity using 1% washed chicken erythrocytes. Out of total of 40 samples, 30 samples were found HA positive. All the HA positive samples and samples found negative were processed for extraction of viral DNA with Genomic DNA purification kit. The net isolated DNA samples were subjected to Polymerase Chain Reaction (PCR) by using specific primers (developed against the variable genomic region of the capsid protein of the DNA) of the CPV. One of the selected primer pair designated as (37, 38) amplified the genornic region present in the CPV-2, the other primer pair designated as (39, 40) that amplified the region present in the CPV-2b.Optimization of PCR was done for the molecular level diagnosis of canine parvovirus and this technique was previously not available or standardized in local conditions of Pakistan. With the use of primer pair (37, 38) best results were found on following optimizing conditions like annealing temperature 55°C, primers concentration (reverse and forward) 25pmoles, Magnesium Chloride concentration 2mM, DNA(Template) volume Spl, Taq DNA polymerase(12.5U) and 2001iM of each dNTPs. For the primer pair (39,40) best results were found on following optimizing conditions like annealing temperature 5 5°C, Magnesium Chloride concentration 1.5mM, primers concentration (reverse and forwid) 20pmoles, DNA(Template) volume 6il, Taq DNA polymerase(12.5U), 2001.iM of each dNTPs. The PCR products were analyzed and compared for their banding pattern of 681bp and 427bp amplified by the primer pairs (37, 38) and (39, 40), respectively, against the standard DNA ladder (1 OObp) by using horizontal agarose gel electrophoresis. One of the HJA positive samples was not amplified by the PCR. The results of this study showed that the specificity of PCR reaction as compared to the HA test and the presence of the CPV-2 and 2b in Pakistan as the prevalent pathogenic strains. This study has been a first step for the molecular characterizationldiagnosis of canine parvovirus in local conditions of Pakistan. It is hoped that this study will pave the way for further advanced studies on this topic.
Availability: Items available for loan: UVAS Library [Call number: 0972,T] (1).
215.
Immune Response Of Buffaloes To Foot And Mouth Disease Virus Vaccine
by Munir Ahmad Tariq | Prof.Dr.Khushi Muhammad | Prof.Dr.Muhammad Akram Muneer | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Nature of contents: ; Literary form: Publisher: 2007Dissertation note: Foot and Mouth Disease (FMD) is a highly contagious infection of cloven-footed animals such as buffalo, cattle, sheep, goats and camels and FMD is characterized by high rise of temperature, salivation, smacking of mouth, vesicular lesion in the buccal cavity, inner flares, coronary band and interdigital spaces, memory glands etc. In Pakistan FMI) disease is caused by "0", "A" or "Asia-i" type of the virus of an Aphthovirus of Picornaviridae. The vaccinal serotypes of FMD virus were characterized as "A", "0" and "Asia-i" by virus neutralization test using imported mono-specific rabbit antiserum. Each of the serotypes multiplied rapidly on monolayer of Baby Hamster Kidney -21 (BHK-21) cells.
The BHK-2 I cells were propagated in carrel and roux flasks in MEM 199 containing 10% fetal bovine serum. Heat treated goat serum was equally effective as growth promoter for BHK-21 cell line. The cells rapidly multiplied and formed a monolayer within 72 hours at 37 °C. The cells were harvested using trypsin (0.025%) without affecting the cell viability that was observed by cytometeric as well as by colorimetric assays. The cells were stored in cryogenic containers and revived successfully on 12 months post storage.
The FMD virus isolate ("0", "A" and "Asia-i") grew well on the monolayer of BHK-21 cells and produced more than 106, and i04 units of the Tissue Culture Infective Dose-50 (TCID50) on 5th passage, respectively. Each of the virus serotypes was effectively inactivated using 0.12 % formaldehyde, or 0.004 M of Binary Etyhieneimine (BET).
The inactivated virus suspension was admixed with either oil base, lanolin or aluminium hydroxide gel and homogenized to get stable vaccine preparation. The adjuvant containing vaccines induced detectable level anti-FMDV-VN antibodies titer in buffalo calves on 19 days post-priming. Oil and gel based FMD vaccines induced detectable geometic mean titer (GMT) of the anti-FMDV-CFT antibodies (2-3 and 7-8) on 19 days post vaccination, respectively. The oil and gel based vaccines induced 1: 64 and 1:80 GMT titer of the anti-FMDV-CFT antibodies on 128 and 64 days post-vaccination, respectively and the titer declined there after as 1: 9 and 1: 3.3 on 258 days post vaccination. From this study it can be concluded that oil based vaccine induces the antibody response in buffalo latter than that of gel adsorbed vaccine. Higher titers of the antibodies are retained for comparably longer period of time by oil based vaccines. Moreover, age of buffaloes, animal species and vaccine storage at 4 C exhibited undetectable effects on the antibody response to the vaccine.
The study has indicated that vaccination programs against field infection of FMD in all the domestic cloven footed animal species could be effective way of immunoprophylaxis.
Availability: Items available for loan: UVAS Library [Call number: 0973,T] (1).
216.
In Vitro Production Of Aflatozin (B1), Its Purification, Estimation And Biodegradation
by Muhammad Hanif Khan | Dr. Irshad Hussain | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2007Dissertation note: In this study a toxigenic strain of A. parasiticus was used for the production of the aflatoxin Eli. This strain of the fungus was taken from naturally growing fungal samples such as bread, rice, vegetables, fruits, maize etc. from various locations in and around the university. The unpurified sample was culture-purified on Sabouraud Agar through repeated culture technique. This strain was maintained on Sabouraud agar slants at 4 °C. The purified sample of this fungus was harvested with 0.01 % Tween 80 then inoculated to the autoclaved broken rice. The rice was incubated at 28 °C in a dark room for a period of' 7 days and shaken two times a day. After 7 days of incubation the rice as autociaved at 121 °C in order to inactivate the fungus. The rice was dried and crushed to powder in a blender. The toxins were extracted through chloroform methpd and evaporated to dryness in water bath at 6 °C. The toxins were again dissolved in I ml ehiorokrm and stored at 4 °C for further use in the study. The toxins were then purified on silica gel plate in order to get the AFBI only through comparison with the AHII standard. 'l'he aflatoxin BI band was scratched from the silica gel plate and subjected to centrifugation in order to remove the silica gel. The purified Bi toxin was then dissolved in chloroform for further use. The AFB1 was then identified and qilani i lied through the fol lowing three methods namely,
1)High perlirmance Liquid chromatography
2) Thin layer chromatography
3) Spectrophotornetry
The sensitivity of the above three methods was determined and compared which was 1 .3 ng/ml for HPLC. 6.3 ng/ml forTLC and 40 ng/mI for Spectrophotometry. The purified AFBI was then estimated by HPLC the amount of which was 238 ± 9.8 jig / ml. however the toxins estimated through TLC were 127.8 ± 24 jig/mi and spectrophotometry were 26 ± 4jig/ml ml. The HPLC method was proved to be the most sensitive method for detection and estimation of the aflatoxin as compared to the other two methods. The HPLC, estimated toxins were then checked for their biodegradation for this purpose the AFBI were kept at three different temperatures and dissolved in three different solvents. As the toxins had previously been dissolved in the chloroform so 400 jil of the chloroform was evaporated to dryness in order to dissolve them in their respective solvents. The toxins were dissolved in 400 j.tl of chloroform, acetone and methanol and kept at 25, 4 and -20 °C. These toxins were then stored for a period of four weeks and the tested for their biodegradation after four weeks by HPLC.
The temperature at which the toxins showed less degradation was determined. That temperature at which the toxins showed less degradation was - 20 °C and the amount of the toxin at this temperature after four weeks of storage was 197 jig/ml. The solvent in which the toxins exhibited less degradation was acetone at all the three temperature conditions (25, 4 and -20 o() I lowever chloroform at 25 °C was exhibiting less degradation as compared to the other two solvents. Methanol was proved to be the least good solvent because the toxin was showing degradation in this solvent. The acetone at -20 CC was the most appropriate solvent for toxins storage.
In second experiment the fresh toxin was dissolved in acetone and tested fbr their degradation on weekly basis by HPLC. The toxin level at the beginning of the experiment was 447 ± 9 ug/mI which when detected after a weak was 398.67 ± 9.8 jig/mI. The same toxin when tested after 2 weak period storage exhibited a level of 295.9 ± 20 ug/mi followed by 265.7 ± 3Ojig/ml. after the end of the third weak indicating that there was an appreciable level of toxin degradation. The toxin tested on the fourth week of storage were 267± 31 ig/ml which exhibited no degradation as compared to the degradation of the third week.
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217.
Electrophoretic Profile Of Escherichin Coil From Calves Suffering From White Scour
by Kaleem Ullah | Prof.Dr.Irshad Hussian | Prof.Dr.Masood Rabbani | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2007Dissertation note: Mortality in neonatal calves has mostly been attributed to infectious agents that generally result in diarrhea, dehydration, and death causing huge economic losses in livestock production. Colibacillosis or white scours is the most important disease of bovine calves. The most common enteropathogen which causes diarrhea in new born calves is the Escherichia coli. Given the range of ill- effects of Escherichia coli causing diarrhea in new born calves in Pakistan there is strong need for detection of strain involved.
In the present project the relatedness by electrophoretic profile of Escherichia coil isolates involved in white scour at a dairy farm were studied, to compare the isolates of Escherichia coli from different dairy farms. The 25 samples from clinically affected calves at farms were collected, for isolation of Escherichia coil these samples were streaked onto MacConkey Agar media plates and Escherichia coil were confirmed through biochemical (IMViC) and sugar fermentation tests. These confirmed Escherichia coli isolates were subjected to sodium dodecyl sulphate polyacrylarnide gel electrophoresis (SDS-PAGE) for electrophoretic protein profiling. The banding pattern various isolates as against known molecular weight markers were recorded for interpretation of results.
SDS-PAGE of whole cell protein extracts of E. coil strains produced patterns containing about 25 discrete bands with molecular weights of 16000 to 250000 Da. The whole cell protein patterns of E. coil isolates were fairly homogenous with some
Veraibility primarily localized in the low molecular weight region estimated molecular weight. 20000 to 60000 Da. The most common peptide bands were band no. 13, 14, 15.
16, 17 and 1 8. detected almost in all strains isolated. In order to estimate relationship among E coli isolates a genetic distance was constructed with similarity matrix using Bio-Rad (Quantity one) software. A complete linkage dandrogram was constructed by Dice coefficient analysis using Quantity One Software. The dandrogram divided the isolates into four clusters based on the whole cell protein profile. The first group contained isolates 2. 4. 6, 7, 3, 5, 13 and 8 that shared 52 percent similarity. The second cluster included isolate numbered 12, 18, 9, 16 and 17 at 60 % similarity. The third cluster included isolate numbered 19, 20 and 15 with 57 percent similar band pattern. The fourth cluster included isolates 10 and 14 that shared 59% similarity.
It was concluded that protein profile is one of the useful methods for determining the relatedness or unrelated ness of bacterial strains. As immunization has been often attempted in efforts to control the calves' diarrhea, the diversity of E. coil strains and difficulties in vaccine development instruct that use of one or likely several strains of E.coli that are antigenicallv representative of the majority of the causative strains in a herd or even in herds located in the same geographical region are recommended. Decision on representative strains can be made on basis of whole cell proteins, the isolates could be characterized by their total protein profiles and use of one or likely several strains of E. coli that arc antigenically representative of the majority of the causative strains will overcome vaccine development difficulties.
Availability: Items available for loan: UVAS Library [Call number: 0979,T] (1).
218.
Factors Affecting Hemagglutination Potential Of Avain Influenza Viuruses (H5, H7, H9 Subtypes)
by Mubashir Hussain | Prof. Dr. Muhammad Akram Muneer | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2007Dissertation note: The objective of this study was to standardize hemagglutination and hemagglutination inhibition tests for AIV H5, H7 and H9 subtypes. These subtypes were propagated in 09-day old chicken embryonated eggs and after 72 hours post incubation the allantoic fluid (AF) was harvested and confirmed by spot agglutination test and by AGPT. While standardizing HA test maximum titers were recorded using 1% RBCs of chicken, human blood group Qe and dog using phosphate buffer saline (PBS) as a diluting agent for washing suspension of erythrocyte and by incubating the micro titer plates at 22c or 37C for 30 minutes or 40 minutes time period. The AIV subtypes eluted rapidly with increase in temperature with maximum elution observed within the time period of 8 hours. The live AIV provided much higher HA titer when compared with the titers obtained from AJV subtypes inactivated with formalin or Binary ethylene imine (BET). The BET was found to have little effect on HA activity as compared to formalin. While standardizing the HI test the best titers were obtained using 4 HA units of AIV antigen as compared to 1 HA and 8 HA units of antigen and by incubating the micro titer plates for 60 minutes period (time given for antigen-antibody reaction before the addition of erythrocytes suspension).
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219.
Comparative Efficacy Of Passive And Active Immunization During Newcastle Disease (Nd) Outbreak In Broilers
by Mushtaq Ahmad Gondal | Prof.Dr.Irshad Hussain | Prof | Prof.Dr.Khushi Muhammad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2007Dissertation note: Newcastle disease is an economically important disease of poultry resulting in huge economic losses every year to the poultry farmers in Pakistan. To compare passive immunization and active immunization during outbreak of Newcastle disease a total of 140 chicks at 16th day of age were divided into seven groups (A, B, C, D, E, F and G) containing 20 birds in each. The level of maternal antibody in chicks, was determined by haemagglutination inhibition titres which revealed that it was the highest at one day and decreased with increasing age. Newcastle disease virus gifted from Dr. Shafqat Fatima Rehmani, Director, Poultry Vaccine Center, Karachi, Sindh was pathotyped by using MDT and ICPI. The Embryo Lethal Dose5o was calculated to be 1083h/0. imi and was found highly pathogenic. Infection was induced in birds through administrating 100 ELD5O1O631/0.lml of Velogenic Newcastle disease virus. Birds of group A, this group seved as a negative control. In group B, this group acted as a positive control. Infection was given by using 0.1 ml of 100 ELD50 of Velogenic Newcastle disease virus intranasally at 26 days of age. In group C, at 16th days of age, all birds in this group were vaccinated with Newcastle disease virus vaccine. At 26th day of age, this group was exposed to infection as mentioned above. In group D, at 24th days of age, all birds in this group were vaccinated with Newcastle disease virus vaccine. At 26th day of age, infection was given by using 0.1 ml of 100 ELD50 of VNDV intranasally into individual bird.
In group E, infection and vaccination were given simultaneously at 31 day of age. In group F, In this group, infection was given by using 0.lml of 100 ELD50 of VNDV intranasally, at 26 days of age. When Newcastle disease symptoms were noticed, birds were vaccinated with lentogenic strain of Newcastle disease virus vaccine (Lasota, TAD- Germany) by using 0.5mllbird orally.
In group G, infection was given by using O.lml of 100 ELD50 of VND.V intranasally, at 26 days of age. When Newcastle disease symptoms were induced, birds were treated with 64 units of anti-NDV-haemagglutination inhibition yolk antibodies. Use of Lasota vaccine and preformed antibodies in yolk help in decreasing economical losses due to outbreak of Newcastle disease in poultry.
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220.
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: ; 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.
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221.
Electrophoretic Profile Of Normal And Hydropericadium Virus-Infected Liver
by Shahid Khan | Prof.Dr.Irshad Hussain | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2007Dissertation note: Hydropericardium syndrome primarily affects the broilers between the ages of 2-7 weeks. 1he 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 check the presence of immunogenic proteins in commercially available HPS vaccines Hyper immune serum was raised in 60 (3 weeks old) broiler birds using different commercially available HPS vaccine. The sera were subjected to AGPT for screening out HPS positive serum samples. The HPS infected livers were homogenized and loaded in alreadv prepared agar gel plates. The plates were incubated for 48 hours at room temperature in humid plastic box. Only 5 liver samples were found positive .The HPS autogenous vaccines,HPS positive liver samples normal liver samples were subjected to homogenization, sonication, chloroform treatment, ammonium sulphate precipitation, mixed with equal volume denaturing buffer and then boiled for 3 minutes to extract total proteins of Sample. . Large variations in the protein loading of samples in adjoining lanes lead to distortion.
Spectrophotometer determination of protein concentration assay were used to quantitate the known protein samples (bovine serum albumen is commonly used standard or this method) to standardize protein concentration. A28o were used to determine protein concentration and a calibration curve was created by plotting and performing regression analysis of A28o versus concentration of the standards and absorbance of the sample were used to determine the concentration from the calibration curve by using the formula. (concentration =Standard Factor* Dilution Factor* Optical Density.
Total protein analysis was carried out by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting technique.
A typical gel of 9% acrylamide composition was used to nicely separate polypeptides of samples to analyze the entire profile of a fraction that contains heavy and light polypeptides. Best results were obtained when 30 pi of a 20-30 mg/mi final concentrations of denatured protein sample were loaded per sample well .Each sample was repeated twice on gel. The gel was cutted at the centre vertically. 0.1% Coomassie Blue dye in 50% methanol, 10% glacial acetic acid were used to stain half of the gel for detecting protein while half of the gel was subjected to western blotting. Relative molecular weight (MW) of each protein fraction was determined by plotting a standard curve. The western blot analysis of proteins of hydropericardium syndrome virus infected liver and HPS autogenous vaccine, separated on 9% gel showed one immunogenic protein, molecular weight 15-20 kDa. However, further studies are needed to establish its immunogenic nature and feasibility for its use as vaccine.
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222.
Sero-Prevalence Of Brucellosis In Buffaloes And Cattle Of Swat Valley And Government Livestock Farms,Nwfp
by Azhar Khan | Prof.Dr.Masood Rabbani | Prof.Dr.Azhar | Prof.Dr.Khushi Muhammad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2008Dissertation note: Brucellosis is an infectious zoonotic disease that is associated with chronic debilitating infections in humans and reproductive failure in domestic animals (Corbell, 1997). The sero-prevalence of brucellosis in buffaloes and cattle were undertaken by collecting samples from animals brought to various slaughterhouses and Private and Govt. farms in Swat valley and Peshawar division by screening through i-LLISA, MRT and RBPT. Out of 850 samples, 600 sera, 200 milk samples were collected along with 50 samples of slaughterhouse worker, butcher and veterinarian for this study.
All the serum samples tested through RBPT and I-ELISA showed the overall prevalence 3.67% and 4.33% in the cattle and buffaloes population respectively while the combined prevalence in the cattle, buffaloes and human population through RBPT was 3.38 % and through i-ELISA was 4%.
The high rate of brucellosis was recorded through RBPT and i-ELISA in buffaloes
( 4.75%,5.5%) while 0.0% prevalence in male buffaloes through RBPT and iELISA, where as in female buffaloes it was 4.85% through RBPT and through i-ELISA was 5.626%.
The comparatively low rate (1.5%) of brucellosis was noted in cattle through the RBPT and 2% through i-ELISA while in female cattle it was 1.587% through the RBPT and through i-ELISA 2.12% with 0.0% in males.
Among the serum samples (30) of buffalo and cattle having reproductive disorder were tested through the same tests which showed overall prevalence 16.6%. The prevalence at Cattle Breeding and Dairy farm 1-larichand and Livestock Research and Development Farm (Surrezai was 0.0% through Milk Ring Test and i-ELISA.
Also cattle milk samples (110) from private farms in swat valley showed prevalence through Milk Ring Test 0.9% and through i-ELISA prevalence was noted to be 1 .82%. As 50 human serum samples were tested through RBPT and i-ELISA but none of these samples were positive showed that the prevalence of brucellosis in human being is very low, The comparison of RBPT, MRT and i-ELISA (milk and serum) was also analyzed statistically by z-test, the data revealed insignificant results.
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223.
Effect Of "In Process Quality Control "Factors On Efficacy Of Bird Flu Vaccine
by Saeed Khan | Prof.Dr.Khushi Muhammad | Prof.DR.Irshad Hussain | Prof.Dr.Muham | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2008Dissertation note: Bird Flu virus was recovered from lungs, trachea, spleen and fecal contents of the infected birds in 10 days old chicken embryos. HA activity and biological titer of the virus improved by serial passages in the 10 days old chicken embryos. This could be due to high rate of mutation of the Bird Flu virus with its successive passages. Formaldehyde and binary ethylenimine (BET) effectively inactivated the virus. However formaldehyde inactivated virus showed mitigation in HA activity during storage. The BET 5mM inactivate the virus with in 16 hours of incubation at ambient temperature (25°C) or 37°C. It has minimal detrimental effect on the HA activity of the virus, even during storage at refrigeration temperature.
Bird Flu virus vaccines without adjuvant induced poor antibody response in the vaccinated broilers. The vaccine containing aluminium hydroxide gel induced antibody response that reached at peak level on 1 8 days post priming and decline thereafter. The vaccine containing montanide (oil based vaccine) increased (90.5 GMT) up to 42 days of age. Boosting of the birds primed with gel based Bird Flu virus vaccine improved the production of antibody titer, while boosting of birds primed with oil based Bird Flu vaccine showed undetectable effect. This was due to increasing trend of antibody titer in oil based primed birds. Montanide based vaccines are therefore recommended for broiler, layers and breeders in high risk area of the disease.
Vaccines containing decreased infectivity titer induced decrease antibody titer in the vaccinated broilers. Bird Flu virus improved its HA activity and infectivity titer with serial passages in 10 days old chicken embryos. It is worth mentioning that serial passages of the virus tremendously decreased its antigenicity. It is therefore recommended to prepare commercial vaccine fom passage number 1-4, for effective immuno-prophylaxis.
The Bird Flu virus (H5N1) mutates very rapidly every time it passes through chicken embryos. It is therefore suggested to grow the virus at least in bio-safety level-ll plus (BSL-II +) laboratories.
On account of it high rate of mutation and risk of human health hazards it is suggested that Bird Flu virus (H5N1) may not be used in vaccine production, however other serotypes containing H5 and N antigen other than N1 may be used for production of commercial vaccine.
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224.
Effects Of Infectious Bursal Desease Vaccine And Vaccination Schedule In Imunity Induceds By Newcastle Disease
by Kashif-ur-Rehman | Prof.Dr.Mansur-ud-Din Ahmad | Dr.Imran Najeeb | Prof.Dr.Azhar Maqbool | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2008Dissertation note: The study was carried out for verifying the interference of infectious bursal
disease vaccines conmonly used in Pakistan on the immunity to Newcastle disease vaccines. Infectious bursal disease (IBD) is a viral disease producing suppression in humoral immune response causing degeneration of bursa of Fabricius. Different vaccines are available in the market for mass scale immunization of chickens. The study was carried out to compare the immunosuppressive effects of intermediate strain, hot strain and complex IBDV vaccines on immunity induced by NDV vaccine. Vaccine efficacy was studied by measuring the induced humoral antibody level using HI (Haemagglutination inhibition) test for NDV and indirect ELISA (enzyme-linked immunosorbent assay) (Kirkegaard' & Perry Laboratories - KPL) to detect antibodies against IBDV.
The parameters used to evaluate the effects of IBDV vaccine on broiler chicks were immune response to NDV vaccination, weight of lymphoid organs such as bursa of Fabricius, thymus and spleen, post virulent NDV challenge and FCR. The results showed that IBDV vaccinated groups A, B, C, D and E had lower HI antibody profile, higher bursa, spleen and thymus body weight ratio, poor FCR and higher post challenge mortality than NDV vaccinated group F. The HI serum antibody profile revealed that the groups vaccinated with IBDV hot strain had significantly lower antibody titer as compared to the intermediate strain of IBDV vaccinated birds. In addition to that hot strain vaccine found to be more damaging to the bursa, spleen and thymus than the intermediate strain vaccine. The hot strain had adverse effects on the feed conversion ratio of birds as well. The challenge with virulent NDV revealed that IBDV vaccine treated groups were overall higher mortality than the only NDV vaccinated chickens.
The study suggested the use of intermediate strain as vaccine since it had least interference with the antibody production against ND. However, hot strain used in this study had adversely affected the NDV HI. titer and caused more damage to the lymphoid organs and reduced feed conversion ratio (FCR).
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225.
Comparative Neutralizing Effect Of Mycotoxin Binders And Its Role In Improving Humoral Immune Responses
by Hazrat Nabi | Prof.Dr.Irshad Hussain | Dr.Aftab Ahmad Anjum | Prof | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2008Dissertation note: µPoultry industry in Pakistan provides inexpensive meat for almost everybody, rich or poor. Poultry farming which once was a blooming business now faces many challenges. Although, infectious agents constitute a major challenge for this industry, mycotoxins are also considered a threat for poultry business worldwide. Mycotoxins, especially aflatoxins are very common in the feed stuff used in the assembly of poultry feeds. The ill-effects associated with aflatoxins are well documented. There are toxin binders marketed by various companies who trumpet various beneficial effects of using these products in poultry. The present project was undertaken to corroborate their claims and also to see whether the use of such products may not result in ill effects that might impact negatively on various production parameters of poultry.
Two commercial products (Mycotox® and Mycofix® Plus 3.0) were used to study their contribution on the immune response against ND, feed consumption, weight gain, feed conversion ratio and "lymphoid organ body weight ratio" in broiler chickens. Asp ergillus parasiticus was used for the production of the aflatoxin which was fed to various groups of birds.One hindered and eighteen day-old broiler were randomly divided into 6 groups viz. A, B, C, D,E and F each comprising 18 chicks. The chicks in each group were further randomly sub-divided into 3 replicates comprising of 6 chicks in each. Group A= Only Feed (Negative Control for AF),Group B= AF @150 µg/Kg of feed. (Positive Control for AF), Group C Mycotox® @ 1 gum/Kg 'feed (Positive Control for Mycotox®), Group D= Mycofix® Plus 3.0 @ 2.5gm/Kg of feed. (+ Control for Mycofix® Plus 3. Q), Group E= AF 1 50 µg/Kg and Mycotox® @ 1 gm/Kg of feed (Experimental Group for Mycotox®), Group F AF @ 150 µg/Kg and Mycofix® Plus 3.0 @2.5gm/kg of Feed (Experimental Group for Mycofix® Plus 3.0).
The rice powdered material containing AF was mixed according to the calculation to get desirable level of AF (150 µg/kg) in the feed. The birds were vaccinated against Newcastle Disease at the age of 6th days (Intra-ocular route) and then at 24th day (drinking water) and were d libitum. Effects of adding mycotoxin binders to feed containing 1 50ppb aflatoxin were in broiler chicks from 14 to 42 day of age. Compared to the B (positive control for aflatoxin) fed aflatoxin alone significantly reduced geometric mean titres, feed consumption, body weight gain and feed conversion ratio. However, no differences in GMT, body weight gain and feed conversion ratio were found between the chicks fed mycotoxin binders C (positive control for Mycotox®) and D (positive control for Mycofix® Plus 3.0) or mycotoxin binders plus aflatoxin treatment groups E (Experimental Group for Mycotox®), F (Experimental Group for !vlycofix® Plus 3.0) and the control group A (Negative control for Aflatoxin), indicating apparent protection against the deleterious effects caused by aflatoxins. Treatment related changes in organ body weight ratio of thymus, spleen and Bursa of fabricius were also observed. Most of the parameters measured for the birds fed mycotoxin binders did not alter. The addition of mycotoxin binders to aflatoxins contaminated feed diminished the adverse effects of aflatoxins on antibody titres against Newcastle and most relative organ weights. These findings suggested that mycotoxin binders can effectively reduce the toxicity of aflatoxin in broiler chicks and mycotoxin binders can be potential ameliorator against aflatoxicosis in broiler chicks.
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226.
Study On Molecular Diagnosis Of Canine Distemper Virus
by Muhammad Zubair Shabbir | Prof.Dr.Masood Rabbani | Prof.Dr.Khushi Muhammad | Prof.dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2008Dissertation note: Samples from fourty five dogs were submitted to the University diagnostic Laboratory, University of Veterinary and Animal Sciences, Lahore from January, 2007 to January 2008 for diagnosis of CDV infection. These dogs presented to referring veterinarians with clinical signs suspicious of CDV infection. Hematological examination (lymphocyte count) was carried out using K-EDTA anti-coagulant added whole blood and RT-PCR tests were performed using biological fluid samples that include plasma, nasal and conjunctival swabs. Only distemper positive dogs by RT-PCR were followed up for subsequent lymphocyte count and prognosis of distemper infection.
All the distemper positive dogs were lymphopenic but the degree of severity was variable as the samples were collected from dogs of different ages and phase of the disease. The study revealed that lymphopenia can be used to support presumptive clinical diagnosis but required laboratory procedure for confirmation and animal regain its normal value with the passage of time subjected to recovery. During followed up, two dogs were found to be dead because of CDV infection mixed with secondary bacterial infection in which one exhibited the nervous sign like teeth grinding, ataxia, convulsions and in coordination in body movements.
Only ten (22.22%) samples were found positive by RT-PCR using plasma, nasal and conjunctival swabs. CDV RNA was detected in 60% of plasma samples, 70% of nasal and 100% of conjunctival swab sample from lymphopenic dogs whereas the percentage was 13.33, 15,55, and 22.22 from a total of 45 samples. No amplicon of expected length was obtained from normal healthy dogs. On comparison of different fluid samples, the sensitivity of conjunctival swab was found to be highly significant followed by nasal swab and plasma.
In conclusion, Lymphopenia is the suggestive of clinical infection of dogs with canine distemper virus ad can help in presumptive diagnosis. It is not necessary that all lymphopenic dogs are distemper posit it requires further laboratory confirmtion. In this context, RT-PCR is test of choice with samples including conjunctival swabs and plasma.
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227.
Charecterization Of Peste Des Petits Ruminants Virus (Local Strain) From Small Ruminants
by Sher Bahadar Khan | Dr.Aftab Ahmad Anjum | Dr.Azhar | Dr.Irshad Hussain | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2008Dissertation note: The objectives of the study were to isolate PPR virus (Local strain) using Vero cell line, Identification of PPRV through Cytopathic effects (CPE) produced by PPRV on Vero cell line. To detect haemeagglutinability of PPR virus with RBCs of poultry, duck, goat, pigeon, sheep, horse and human through Haemeagglutination test. And confirmation of PPR virus through Hemeagglutination inhibition test and Immunocapture Enzyme Linked Immunosorbent Assay (Ic ELIZA). For this purpose 120 tissue samples (40 Necrotic debris in buccal mucosa, 30 each nasal and ocular discharges and 20 lymph nodes) were collected from clinical positive cases. These samples were moistened with 2-3 drops sterile PBS and were brought immediately to the laboratory in sterilized universal container under refrigeration temperature. The tissue samples were processed for virus isolation. After sterilization of the glassware, Dehydrated modified Essential Medium (DMEM) was prepared according to the manufacturer's instructions for cultivation of Vero cells. Stock solution of phenol red, Carbonate/bicarbonate buffer, Stock antibiotic solution, Trypsin, Versene solution and Trypsin-versene (TV) solution were prepared. The inoculums of Pestes des Petitis ruminants virus was filtered through 0.2um pore sized syringe filter (Millipore,USA) and transferred to a pre-sterilized McCartney bottle.When a complete monolayer of the Vero cell line was formed and almost 80 % confluency was obtained, the exhausted medium from the carrel flask was discarded and new filtered and sterile maintenance medium (25 ml) was added per flask. The virus inoculum was inoculated on Vero cell line and examined daily for CPE. The haemeagglutinability of the virus was checked with RBCs of chicken, duck, pigeon, sheep, goat, horse and human blood group 0. The The haemeagglutinability of the virus was also checked under different conditions i.e. influence of diluents, influence of temperature and influence of incubation. Finally Haemeagglutination inhibition test and ic ELISA was performed.
Availability: Items available for loan: UVAS Library [Call number: 1039,T] (1).
228.
Selection Of Thermostable Reconstituted Newcastle Disease Virus Progeny From Vaccinal Strain(La Sota)
by Faiza Ghazanfar | Prof.Dr.Mansur-ud-Din Ahmad | Prof.Dr.Mohammad Akram Munir | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Nature of contents: ; Literary form: Publisher: 2008Dissertation note: Poultry sector is the second largest industry after textile in Pakistan. It is threatened by various diseases. Every year, Newcastle disease is causing enormous losses to this industry. A thermostable vaccine of Newcastle disease will be an answer to this problem.
A vaccine strain of Newcastle disease La Sota was divided into two groups BATCH-A and BATCH-B. BATCH-A was selected for sudden heat exposure at 45°C. But virus could not survive the direct exposure of this temperature. So BATCH-B was selected for gradual increasing heat shock treatment. The starting temperature was selected at 40°C for 20 minutes. Stable NDV virus at that temperature was subjected to further heat treatments at 42°C for 20, 25, 30, 35, 40, 45, 50, 55 and 60 minutes. Virus suspension was labeled as "BATCH-B 1".
BATCH-B 1 was divided into four groups for different heat treatments. They were labeled as Bl-A, B1-B, Bl-C and B1-D. B1-A was subjected to the temperature of 50°C for 20 minutes. B1-A titer was 2 (1:32). It showed drop in HA titer upto fourth well (1:16). B 1-B was placed at room temperature for three consecutive days with HA checking after every 6 hours. No drop in HA titer was observed. B 1-C was placed in a bright sunlight for 12 hours and HA titer was dropped down by one well (1 log2). B1-C HA titer was 2 (1 :32).It dropped down to 2 (1:16). Lastly, the parent virus suspension i.e. BATCH-B with HA titer of tenth well (1:1024) and virus suspension of "B 1 -D" with HA titer of fifth well (1:32) were placed in 56°C for 30 minutes, 60 minutes, 90 minutes and 120 minutes as described by King (2006) and Wambura et al (2006).
The virus suspension of "B 1 -D" survived the temperature but the titer dropped to third well (1:8) at 30 minutes. It was dropped further to 21(1:2) at 60 minutes and maintained its titer i.e. 21(1:2) at 90 minutes. The BATCH-B did not show any HA titer at 30 minutes and onwards. The E1D50 of B1-D was calculated as 10-7.4
The study will help to minimize the challenges at farm level. The vaccine can be transported in the rural areas as the thermostable strain which will be able to bear the harsh conditions of weather in Pakistan. Furthermore, its easy application will help the farmer to vaccinate his bird flock against this disease without any worries.
Availability: Items available for loan: UVAS Library [Call number: 1040,T] (1).
229.
Electrophoretic Profile Of Cellular Proteins Of Staphylococcus Aureus From Mastitic Cattle
by Muhammad Zubair Munir | Prof.Dr.Irshad Hussain | Prof.Dr.Masood Rabbani | Prof.Dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2008Dissertation note: Decline in milk production has. mostly been attributed to infectious agents that generally result in swelling of udder, changes in the constitutions of milk and finally induration of the udder which causing huge economic losses in livestock production. Mastitis is most important diseases of all the lactating animals. The most common Staphylococcus aureus which causes mastitis in cattle. Given the range of ill-effects of Staphylococcus aureus causing mammary infection in cows and buffaloes in Pakistan there is strong need for detection of strain involved. Likewise identification of clones with extensive geographic distribution wall provide insight into strain virulence and pathogenesis and also requiring public health intervention such as vaccination and antimicrobial restriction aimed at reducing the spread of the pathogen. In the present project the relatedness by elcctrophoretic profile of Staphylococcus aureus isolates involved in mastitis at dairy farns were studied, to compare the isolates of Staphylococcus aureus from different dairy farms. The samples from clinically/ subclinically affected at farms were collected, for isolation of Staphylococcus aureus, these samples were streaked onto Staph- 110 agar media plates and Staphylococcus aureus were be confirmed through biochemical and sugar fermentation tests. These conformed Staphylococcus aureus
Isolates were subjected to sodium dodecyl sulphate polycrylaniide gel electrophoresis. (SDS-PAGE) for electrophorctic protein profiling. The banding pattern of various isolates as against known molecular weight markers were recorded for interpretation of results.
It was concluded that protein profile is one of several methods for determining the relatedness or unrelatedness of bacterial strains. As immunization has been often attempted in efforts to control the mastitis, the diversity of Staphylococcus aureus strains and difficulties in vaccine development instruct that use of one or likely several strains in a herd or even in herds located in the same geographical region are recommended. Decision on representative strains can be made on basis of whole cell proteins, since such proteins of Staphylococcus aureus strain has already been shown, the isolates could be characterized by their total protein profiles and use of one or likely several strains of Staphylococcus aureus that are antigenically representative of the majority of the causative strains will overcome vaccine development difficulties.
Availability: Items available for loan: UVAS Library [Call number: 1047,T] (1).
230.
Comparative Efficicy Of Peste Des Petits Ruminant (Ppr) Vaccine S Available In Pakistan In Sheep And Goats
by Muhammad Intizar | Prof.Dr.Manur-Din-Ahmad | Dr.Aftab Ahmad Anjum | Prof.Dr.Azhar | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Nature of contents: ; Literary form: Publisher: 2008Dissertation note: The present study was designed to evaluate the physical factors affecting the PPR 'vaccine and also to compare the efficacy of the locally available PPR vaccines in Pakistan in sheep and goats.
The current study was conducted on 120 different small ruminants ((60 sheep and goats). The humoral immune response was monitored by measuring the antibodies titre through Hl and AGID test. HI and AGID test are reliable and effective methods of diagnosing viral diseases and to evaluate the humoral immune response.
It was concluded that the vaccine should he stored either at -20°C or at 4 C.The vaccine stored at 27 C had a drop in HA titre and no HA activity was found at 40 C
To evaluate the HA activity of PPR virus it is better to use chicken or human group'O' R.B.Cs in a concentration of 1%. The diluent should have the pH 6.8- 7.0.
By evaluating the vaccine efficacy in sheep it was found that after 14th day of vaccination there was a gradual increase in the antibodies titer till the 56th day of vaccination. The locally manufactured vaccine was having a geometric mean titre (GMT)207.9 while the (GMT) of Pestivec was 73.3. 63d day post vaccination. In goats the locally manufactured vaccine was having a geometric mean titer (GMT) 147. while the (GMT) of Pestivec was 48.5.63rd day post vaccination. No antibodies production was there in any control group.
It was concluded form the study that locally produced vaccine is equally good and could be used confidently. It will save also helpful in saving the foreign reserves oh the country.
Availability: Items available for loan: UVAS Library [Call number: 1051,T] (1).
231.
Role Of Herbal Polysaccharides As Immunomodulator
by Muhammed Zafar | Dr.Aftab Anjum | Dr.Muhammed Imran Najeeb | Prof.Dr.Azhar.
Material type: ; Format:
print
Publisher: 2009Dissertation note: Compared to the birds in vaccinated groups that were kept without feeding Livol, the sera of the NDV vaccinated birds kept on Livol had higher antibody titers on day 42.amongst various treatment groups the highest Haemagluttination-Inhibtion titers was recorded in group C feed with Livol treated birds as compared to the other groups. The hematological parameters observed i.e. Packed Cell Volume, Hemoglobin and Total Leukocyte Count were higher in group C2 and C3 than in Group Cl, which shows that increasing Livol concentration have increased the above said parameters. The higher concentrations of Livol (Herbal Polysaccharides) have increased body weight gain than the birds fed with low concentrations of Livol (Herbal Polysaccharides). Treatment related changes in body weight, organ body weight ratio of thymus; spleen and Bursa of fabricius were also observed amongst the various groups. The addition of Livol (Herbal Polysaccharides feed to diminished the adverse/immunosuppressive effects of different vaccine on antibody titers against Newcastle and most relative organ weights. These findings suggested that Livol (Herbal Polysaccharide) can effectively stimulate/enhance the body weight gain, immunity in broiler chicks and Livol (Herbal Polysaccharides) can be potential ameliorator against various vaccines and its adverse/suppressive effects in broiler chicks.
Availability: Items available for loan: UVAS Library [Call number: 1065,T] (1).
232.
Isolation And Identification Of Staphylocococcus Aureus And Salmonella From Snack Food
by M.Rizwan Saifullah | Prof.Dr.Mansur-ud-Din Ahmad | Dr.Aftab Ahmad Anjum | Prof.Dr.Muham | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2009Dissertation note: The present study was planned to investigate microbial load in ready to eat foods (snacks) available at various places in and around Lahore. A total of 60 snacks containing 14 sandwiches, 20 pizza and 26 burgers were procured from various retail outlets. The samples collected were carefully packed in a clean sampling bag and processed in the microbiology laboratory at UVAS following standards protocols.Each sample was processed for APC, Coliform count, Staphylococcus aureus count and the presence of Salmonella. The results were, compared with the guidelines published by PHLS (Gilbert et al., 2000) for comparison with the food standards in developed countries like UK.
In the present study results showed that for aerobic plate counts of 21.4% snacks were of satisfactory quality, 22.5% snacks were of acceptable quality and 58.4% were of unsatisfactory quality. For coliform counts revealed that 72.3% snack food sample were of satisfactory quality while 23.6% samples were of acceptable quality and 4% snacks were of unsatisfactory quality. In the same way Staphylococcus aureus counts for the snack food samples, showed that 22.3% samples were of acceptable quality, 13% samples were of acceptable quality and 64.6% snack food samples were of unsatisfactory quality. However Salmonella could not be detected in any of the sample tested.
In the present study on the basis of aerobic plate counts the unsatisfactory snacks were 64.3% sandwiches, 46% burgers and 65% pizzI. While the coliform counts revealed that 7.1% sandwiches and 5% pizza of unsatisfactory quality.
The results of microbial assessment for the presence of Staphylococcus aureus in snack food indicated that 85.8% sandwiches samples were of unsatisfactory quality, 38% burger samples were of unsatisfactory quality and 70% pizza samples had unsatisfactory microbiological quality. However, no Salmonella spp could be isolated from 60 snack food samples.
Availability: Items available for loan: UVAS Library [Call number: 1067,T] (1).
233.
Effect Of Various Stress Factors On The Immune Response(Ph.D)
by Muhammad Yasser Mustafa Butt | Prof.Dr.Muhammad Akram Muneer | Prof.Dr.Khushi Muhammad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Nature of contents: ; Literary form: Publisher: 2009Dissertation note: Pakistan has vast population of dogs belonging to different breeds. Most of the dogs have no pedigree record which is a great threat to conservation of different breeds. No study on DNA fingerprinting of dogs has been conducted in Pakistan. DNA fingerprinting of dogs is necessary to overcome the problems like forensic cases, sale & purchase, individual identity in case of fertilization by more than one male and ownership disputes. Microsatellite markers have been proved as an efficient and powerful tool for parentage testing and breed characterization of dogs. In this study, a panel of microsatellite markers, having high polymorphism information content (PlC) values, was developed. Blood samples were taken from cephalic vein of two breeds of dogs (German shepherd and Labrador retriever). DNA was extracted by Inorganic method. Primers of microsatellite markers were optimized for successful amplification conditions in the Bio-Rad thermocycler. Multiplex PCR was performed, for amplification of these microsatellite markers on 46 samples belonging to 20 families. Genotyping analysis was performed for the PCR products of microsatellite markers on non denaturing polyacrylamide gel. These results were analyzed statistically software "POPGENE 3.3 and POWER STAT". Allele frequency, heterozygosity, homozygosity, polymorphism information content (PlC), power of discrimination and power of exclusion of all microsatellite markers were calculated. Average power of discrimination among non parents, average hetrozygosity, average observed homozygosity and average polymorphism information content (PlC) value for all alleles was 0.809, 0.6345, 0.29 13 and 0.724 respectively. Moreover combined power of exclusion reached a significant value of 0.9998. Almost all of the microsatellite markers showed significant variations in both German shepherd and Labrador retriever breeds. Microsatellite "REN41D2Ob" showed maximum variation i.e. 17 alleles and microsatellite"REN49F22b" showed the least variation among all microsatellite markers i.e. 4 alleles. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between German shepherd and Labrador retriever breeds.
Results of this study lead to development of a panel of microsatellite markers which can be used for parentage analysis and breed characterization of dogs. This was a preliminary study on dogs in Pakistan. This facility can be provided on commercial basis to pet owners and kennel clubs. Moreover this study can become the basis for further research investigations in canines in Pakistan.
To evaluate effects of various stress factors on immune response and growth performance of broiler chicks, a total of five experiments using 2000 broiler chicks were conducted. In each experiment, chicks were divided into five groups (A, B, C, D and E), and each group consisted of 80 day-old-chicks. In each experiment, the chicks were exposed to stress factors, such as temperature, stocking density, feed deprivation, water restriction and light. Each chick in groups A, B, C, and E was vaccinated against IBV, NDV, IBDV and HPSV, but chicks in group D were kept as unvaccinated controls. Blood samples from each group were collected on 36 day of age, at 18 hrs for determining TLC, DLC and H/L ratio. The antibody titers of chicks in different groups were analyzed using HI test at days 36th and 56. The cellular response was analyzed by injecting PHA-P in the wattles of bird during post stress period. The effects of each stress on lymphoid organs were determined. The potential to resist virulent NDV challenge and effect of stress factors on body weight gains and FCR of chicks was also determined.
In experiment 1, conducted to determine the effect of various temperature ranges on broiler chicks, it was observed that the heat stressed (HS) birds showed non-signilicant dilYerence in TLC values. The HS effect on lymphoid organs indicated that the mean weight of thymus of chicks in group B (0.29±0.02) and C (0.59±0.13) was significantly (P<0.05). lower than those in groups A (4.11±3.26), D (4.50±0.77) and E (4.35±0.21). The mean bursa weight of heat stressed chicks in goups A (0.94±0.59) and B (0.20±0.01) were significantly (P0.05) lower as compared to non- heat stressed chicks in groups D (1.42±0.22) and E (1.33±0.18). The mean spleen weight of groups A (1.21±0.13) and B (1.30±0.11) was significantly (P0.05) lower than groups D (L93±0.16) and E (1.52±0.10) indicating the adverse effect of increased temperature. The FCR valueswere significantly (P0.05) different among groups in 6th1 week and effect of Vitamin C was found significantly (P<0.05) improved than Vitamin E and glucose treatment. At 36 day of age the HI titer was recorded significantly (P<0.05) lower in group A (GMT 61) than B (GMT 144) and C (GMT 109) groups while group E (GMT 186) showed significantly (P<0.05) higher HI titer. At the age day 56 (06 days post challenge) the HI antibody titers in all groups registered a rise except in group D. All the chicks in group D died indicating clinical signs of Newcastle disease. The post challenge GM HI titers recorded in groups A, B, C and E at the age of 56 days was 79, 156, 122 and 216, respectively. There was nonsignificant (P>0.05) differences in wattle thickness (cm) among groups A (1.51+0.06), B (1.77±0.26), C (1.2±0.25) and D (1.52±0.22) but increased in group E (1.83+0.08). The mortality was found significantly (P<0.05) higher in groups A (14) and D (40) on challenge with NDV virulent virus.
In experiment 2, conducted to determine the effect of various levels of stocking densities on broiler chicks, it was observed that stressed birds had showed non-signilicant (P>0.05) difference in TLC values. The effect on lymphoid organs found that the mean thymLls weight (grn) of chicks in groups A (0.37±0.04) and B (0.74±0.17) were significantly (P<0.05) lower than groups C (1.50±0.35), D (4.43±0.72) and E (4.40±0.23). The mean bursa weight (gm) of groups A (0.33+0.03) and B (0.57+0. 1 7) were significantly (P0.05) lower than those of groups D (1.76±0.05) and 13(1.33±0.08) indicating that less space effect the bursa development in the chicks. The mean spleen weight (gm) of groups A (1.18±0.07) and B (1.52±0.20) was significantly (P<0.05) lower than group D (2.37±0.28). The FCR values were sign ilicantly (P<0.05) different among groups in 6thi week and there was non-signiflcant (P>0.05) difference among groups with treatment of Vitamin C, Vitamin E and glucose. At 36th day of age the I-Il titer was recorded significantly (P<0.05) lower in group A (GMT 67) than groups B (GMT 9.6) and C (GMT 102). The chicks in group D (GMT 07) showed negligible HI titer while group E (GMT 185) showed significantly (P<0.05) higher HI titer. At the age day 56 (06 days post challenge) the 1-Il antibody titers in all groups registered a rise except in group D. All the chicks in group D died indicating clinical signs of Newcastle disease indicating that the titers in the birds were not enough to resist the virulent challenge. The postNDV-challenge GM HI titers recorded in groups A, B, C and Eat the age of 56 days were 86, 121, 132 and 210. There wa non-significant (P0.05) differences in wattle thickness (cm) in groups A (1.44±0.07) and C (1.43±0.10) while group E (1.64±0.31) showed significantly (P0.05) increased wattle thickness. The mortality was found significantly (P0.05) higher in groups A (25) and D (40) on challenge with NDV virulent virus. This indicated that less floor space decreased the immune response of the birds which leads to the infection/death.
In experiment 3, effect of feed deprivation at different time intervals on broiler chicks was studied, it was observed that feed deprivation stressed birds had showed non-significant (P>0.05) differences in blood cells population. The effect of feed deprivation on the mean thymus weight (gm) of chicks in group C (0.96±0.29) was adversely affected as the chicks in this group had significantly (P<0.05) lower weight than groups B, 1) and E. The bursa mean weight (gm) of groups A (0.48±0.11) and C (0.52±0.06) was significantly (P0.05) lower than those of groups D (1.40±0.18) and E (1.28±0.11) indicating, that 24 hrs and morning off-feed effect the bursa development in the chicks. The mean splen weight(gm) of groups A (I. 19±0.07) and C (1.21±0.06) vcre significantly (P0.05) lower than groups D and E indicating adverse effect ol24hrs and day off feed on chicks lead to infection. The FCR values were significantly (P0.05) different among groups in 6hhl week and there was significant (P<0.05) differences among groups with treatment of glucose than Vitamin C and Vitamin E treated. groups. At 36th day of age, the HI titer was recorded significantly (P<0.05) lower in group B (GMT 15) than C (GMT 74) and group D (GMT 07) showed negligible HI titer while group E (GMT 140) showed significantly (P<0.05) higher HI titer. At the age day 56 (06 days post challenge) the HI antibody titers in all groups registered a rise except in group D. All the chicks in group D died indicating clinical signs of Newcastle disease. The post challenge GM HI titers recorded in groups A, B, C and Eat the age of 56 days were 68, 54, 115 and 165. There was non-significant (P0.05) difference in wattle thickness (cm) among groups while group E (I .78±0.06) showed significantly (P<0.05) increased wattle thickness. The mortality was Found significantly (P<0.05) higher in Groups C (12) and D (40) on challenge with NDV virulent virus. This indicated that 24 hrs off feed decreased the immune response of the birds which leads to the infection.
In experiment 4, studied the effect of water restriction at different time intervals on broiler chicks, it was observed that water restricted birds had nonsignificant (P0.05) differences in blood cells population except lymphocytes percentage was found higher in groups A (43.7±2.47), C (43.7±1.16) and D (53 .3±1 .30) than group B (39.1±1.06). The effect of water restriction on the mean thymus weight (gm) of chicks in group C (0.60±0.07) was adversely effected as the chicks in this group had significantly (PO.O5) lower weight than group D (4.09±0.70) indicating that increased in the period of water restriction in chicks adversely affected the mean thymus weight and chicks reared on ad-flbituni water had higher mean thymus weight. The mean bursa weight (gui) of group C (0.07±0.02) was significantly (P<0.05) lower as compared to group D (1.37±0.88) indicating that water restriction of 24 hr had affected the bursa development in the chicks. The mean spleen weight (gm) of group C (2.64±1.49) was significantly (P0.05) higher than groups A, B and E. The FCR values were significantly (P<0.05) different among groups in 6th week and there was non-significant (P0.05) difference among groups with treatment of Vitamin C, Vitamin E and glucose treated groups. At 36th day of age the HI titer was recorded significantly (P<0.05) lower in group D (GMT 09) than A (GMT 54) and group E (GMT 109) showed significantly (P0.05) higher HI titer. At the age day 56 (06 days post challenge) the HI antibody titers in all groups registered a rise except in group D. All the chicks in group D died indicating clinical signs of Newcastle disease. The post challenge GM HI titers recorded in groups A, B, C and E at the age of 56 clay was 78, 63, 48 and 134. There was non-significant (P0.05) difference in wattle thickness (cm) among groups B and E and group A (1.28±0.08) showed significantly (P0.05) lower wattle thickness. The mortality was found significantly (P<0.05) higher in groups B (14) and C (21) on challenge with NDV virulent virus. This indicated that 18 and 24 hrs water restriction decreased the immune response of the birds.
In experiment 5, effect of light stress at various time intervals on broiler chicks was studied. It was observed that light stressed birds had showed non-significant (P>0.05) difference on blood cells population except lymphocytes percentage was found higher in groups D (6 1.4±1.16) than group C. The effect on lymphoid organs studied and the mean thymus weight (gin) old chicks in group B (I .90±0.53) was adversely effected as the chicks in this group had significantly (P0.05) lower mean thymus weight than groups D (4.64±0.74) and E (4.34±0.25) indicating that increase in the period of oil-light in chicks adversely effected the mean thymus weight and chicks reared on 24hr light had higher mean thymus weight. The bursa mean weight (gm) 01' group 13(0.43±0.05) was significantly (P0.05) lower as compared to group D (1.59±0.17). The spleen mean weight (gun) of group D (1.88±0.15) was significantly (P<O.05) higher than group B (1.18±0.08). The FCR values were significantly (P().O5) different among groups in 6th week and there was non significant (P0.O5) difference among groups with treatment of Vitamin C, Vitamin E and glucose. At 36° day of age the HI titer was recorded significantly (P<0.05 lower in group C (GMJ 83) and group D (GMT 06) showed negligible HI titer while group E (GMT 128) showed significantly (P0.05) higher HI titer. At the age day 56 (06 days post challenge) in HI antibody titers in all groups registered a rise except in group D. All the chicks in group D died indicating clinical signs of Newcastle disease. The post challenge GM HI titers recorded in groups A, B, C and B at the age of 56 days was 122, 116. 108 and 133. There was non-significant (P>0.05) difference in wattle thickness (cm) among groups while group B (1.53±0.15) showed significantly (P<0.05) increased wattle thickness. The mortality was found significantly (PO.05) higher in groups A (18) and D (40) on challenge with NDV virulent virus. This indicated that 24 hr off-light decreased the immune response of the birds.
Availability: Items available for loan: UVAS Library [Call number: 1080,T] (1).
234.
Antigenic Characterisation Of H9 Subtype Avian Influenza Viruses Isolated Desi And Zoo Birds
by Farrukh Saleem | Dr.Muhammad Mahmood Mukhtar | Prof.Dr.Azhar | Prof.Dr.Khushi Muhammad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2009Dissertation note: Avian influenza is a viral infection which affects mainly the respiratory system of birds. The H7N3 subtype influenza viruses were isolated for the first time in 1994 from breeder flock in northern areas of Pakistan. A second wave of avian influenza outbreak was detected in 1999. The causative agent of this outbreak was H9N2. The H9N2 considered as low pathogenic avian influenza (LPAI) virus and continuously circulating in poultry flocks causing enormous economic losses to poultry industry of Pakistan. This showed that avian influenza viruses are present in commercial poultry. Most of the efforts to isolate avian influenza A viruses are from commercial poultry and these isolations are outbreak based. That is why we have mad’ an effon to isolate and identify H9 subtype avian influenza viruses from apparently healthy live desi and zoo birds (Lahore, Pakistan). We have successfully isolate H9 subtype influenza viruses from these birds during our study.
As these viruses have RNA genome and their RNA polymerase enzyme lacks proof reading activity which resulted in spontaneous mutation in surface glycoproteins (HA and NA) and reassortment of their genomic segments results in escape from host immune response produced by the vaccine. This is the reason that
every year we require a new candidate virus for vaccine preparation. We have made
an effort to isolate and identify avian influenza viruses from live desi and zoo birds of
Lahore and performed antigenic characterization. In this way, we have been able to
know the exact status of avian influenza virus strains present in the desi and zoo birds.
We also have seen that the imported vaccine have less interaction with the local strains and gives less protective titer although it gives best titers when we raise antisera against imported vaccine. The local vaccines although gives a little bit less titer when we raise the antisera against these vaccines but their antisera have more interaction with the local H9 subtype antigen so it gives better protective immune response. By this study we have seen that antisera obtained from infected chicken give more antibody titer as compare to antibody raised in the rabbits. Infected chicken antisera are more reactive as compare to rabbit antisera. This shows that our isolates have highest similarity with the currently circulating viruses.
All above results helped us to devise a new control strategy against avian influenza viral infections present in these local birds. The antigenic characterization of these avian influenza isolates helped us to see the antigenic differences between the isolates of this study and H9 subtype avian influenza viruses used in vaccines. Therefore, this study clearly suggests that a new local H9 subtype avian influenza virus should be used as vaccinal candidate every year for the effective control of influenza viral infections of poultry.
Availability: Items available for loan: UVAS Library [Call number: 1082,T] (1).
235.
Epidemiological Patterns Of Brucellosis In Sheep, Goats And Human Beings
by Jafar Pasand Masoumi | Sh. Muhammad Amin | Dr. Rashid | Mr. Muhammad Naeem | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 1986Dissertation note: The epidemiological patterns of brucellosis were studied on 1554 serum samples (goats, 500, sheep 532, human being 522) and 1027 milk samples (goats 527, sheep 500) for utilizing this information in effectively controlling or eradicating this disease from livestock and human population in Pakistan.
Various tests used comprised slide test, standard tube agglutination test and milk ring test. The antigen for serum agglutination test and slide test was the Standardised Brucella abortus antigen of Veterinary Research Institute, Lahore. For milk ring test haematoxylin stained Brucella abortus antigen developed at Veterinary Research Institute, Lahore was used. An overall incidence of 3.00%, 1.69% and 0.95% was recorded respectively in goats, sheep and human beings by serum agglutination test. The milk ring test gave an infection rate of 9.10% in goats and 7.6% in sheep. The influence of various epidemiolo- gical factors was confirmed and in goats a higher incidence by serum agglutination test was detected in females (3.65%), Teddy breed (.3.93%) and in animals of 4 years and above age group (9.52%). In sheep a higher incidence was recorded in females (3.10%), non-descript breed (3.18%) and in animals of 4 years and above age group (2%). In human beings a higher incidence was observed in males (1.27%), 20-29 years age group (1.86%) butchers (8.33%) and persons in habit of consuming raw milk
(5.40%).
The findings of the present study have revealed an alarming proportion of the disease incidence in our sheep and goats and confirmed the presence of infection in human being. Thjs calls for an immediate response of the experts, and introduction of appropriate brucellosis control measures both in livestock and human population.
Availability: Items available for loan: UVAS Library [Call number: 1116,T] (1).
236.
A Study On The Incidence And Types Of Salmonella Infection In Pigeons
by Salma Kausar | Dr. M. Ajmal | A.R. Rizvi | Sh. Altaf Hussain | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 1984Dissertation note: In the present study faecal material of 100 pigeons, including 50 wild and 50 pet birds, was studied for the presence of Salmonella organisms. The isolation of Salmonella was successful from 21 (21%) samples, Salmonella typhimurium was isolated from all 21 (21%) samples studied. Different biochemical and serological tests were used for the differentiation and confirmation of type of isolates. Serologically "B" group was confirmed. Further work in this field was suggested.
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237.
Seroprevalence Of Brucellosis Animals And Their Role In Transmitting The Disease To The Abattoir Personnel
by Irshad Hussain | Dr. M. Ajmal | Ata-ur-Rehman Rizvi | Dr. M. Sarwar | Faculty of Veterinary Sciences.
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Publisher: 1990Dissertation note: Seroprevalerice of brucellosis--- a disease of great zoonotic potentials and implications----- on 552 cattle, 523 buffaloes, 554 sheep, 524 goats and 87 butchers (working at the abattoir) was studied. Various serologic tests employed for this investigation comprised the slide agglutination test for primary screening and the standard tube agglutination test (SAT) and the enzyme-linked immunosorbent assay (ELISA) for further processing of the sera i.e. quantitation of anti-Brucella antibodies. The slide agglutination test recorded a higher prevalence of the disease whereas both the SAT and ELISA displayed a similar picture of the prevalence. A higher prevalence was noted in buffaloes than cattle while goats outnumbered sheep in this respect.
The prevalence of the disease by agglutination test was found to be 27(4.89), 25(4.527.), 23(4.397.) and 9(10.34X) in cattle, sheep, goats and butchers, respectively, whereas SAT and ELISA registered a prevalence of 18(3.447.), 9(1.627.) and 15(2.067.) respectively buffaloes, sheep and goat!3; however, in human the slide 37(7.07), buffaloes, both the 12(2.177.), in cattle, beings, the
SAT revealed an incidence of 5(5.747.), while the ELISA figured it to be 4(4.59). The influence of various epidemiological factors such as age, sex etc. was proved. Female animals exhibited a higher prevalence (2.97, 3.79, 1.91 and 3.16 per cent in cattle, buffaloes, sheep and goats respectively by the SAT and ELISA) in contrast to the male animals which reflected a lower one (1.41, 1.98, 1.07 and 2.08 per cent, respectively). In cattle and buffaloes above
3-years-age, the prevalnece of brucellosis was recorded to be 2.95 and 4.30 per cent respectively, whereas, sheep and goats above 9 month age demonstrated a prevalence of 2.27 and 3.62 per cent respectively by the SAT and ELISA. Younger animals were all negative.
The enzyme linked immunosorbent assay was found to be a more reliable, sensitive and specific approach than the slide agglutination and standard tube agglutination tests since the former either did not at all react to the sera with a SAT titre of lesser than 1 in 40 or expressed a titre of I in 80 (on a few sera only) which was not of any diagnostic value. The ELISA titres were, on average, about 8 times higher than the corresponding SAT titres.
The results of this study have revealed an alarming prevalence of brucellosis in our animals, which calls for an emergant response of experts for reappraisal and reassessment of the present brucellosis control situation, especially the disease being an important zoonosis and potential threat to the human health.
Availability: Items available for loan: UVAS Library [Call number: 1127,T] (1).
238.
Epidemiolgical Surey Of Rabies By Applying Gel-Diffusion And Its Comparative Efficacy With Flourescent
by Zafar Ul Ahsan | Muhammad Ajmal | Dr. Sh. M. Amin | Dr. Tufail M. Khan | Faculty of Veterinary Sciences.
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Publisher: 1990Dissertation note: An attempt was made to demonstrate rabies virus/ antigen in the saliva/ brain tissue of affected/ suspected/ healthy animals belonging to different species, using agar gel precipitation (AGPT), fluorescent antibody (FAT) and mice inoculation (MIT) to understand the epidemiology of the disease.
A total of 201 dogs (100 stray and 101 suspected), 109 mongooses (102 routinely trapped normal healthy and seven suspected carcases) and eleven suspected domestic animals I six cattle, two buffaloes, two goats and a mule) were tested for the presences of rabies virus/ antigen in the saliva and/or brain tissue. Three diagnostic methods AGPT, FAT and MIT were applied to determine their efficacy. In case of 100 stray dogs, none was found positive. Out of 101 suspected dogs 54 (53.46%) were found positive for rabies. All the routinely trapped mongooses were found negative, where as all the 7 suspected mongooses were positive for rabies. Of the 11 domestic animals six cattle, one buffalo, two goats and a mule were found positive.
Of the total 321 animals of different species 18(5.60%) were shown positive with AGPT, 69(21.49%) with FAT and 71(22.11%) with MIT. The FAT with combination of MIT was found the most sensitive, reliable and quick test for diagnosis.
It was observed that healthy routinely trapped mongooses and healthy stray dogs do not act as carrier. But affected mongooses alongwith rabid dogs also play an important role in disseminating the rabies disease in the country.
Availability: Items available for loan: UVAS Library [Call number: 1128,T] (1).
239.
Preparation And Evaluation Of A Cell Culture Contagious Pustular Dermatitis Virus Vaccine
by Shahida Afzaal | Dr. M. Ajmal | Dr. M. Akram Munir | Dr. T.M. Khan | Faculty of Veterinary Sciences.
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; Literary form:
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Publisher: 1990Dissertation note: The present study was conducted to prepare a cell culture vaccine against CPD to repice the older vaccine so as to induce an effective level of immunity and thus to reduce the economic losses caused by this malady. A local CPD virus strain, isolated from a direased goat during an outbreak occurred at 11.E.S. Bahadurnagar, was adapted on primary lamb kidney cells by giving several successive passages. The isolated virus was confirmed in vivo by known goat pox as well as CI']) viruses and Was used for vaccine production. Confluent primary monolayer of lamb kidney cells was achieved after
5 day of incubation. Viral material after its adaptation on kidney cells at sixth passage was inoculated on monolayer for production of vaccine. When 80% of the cells showed CPE after 6-8 days of post inoculation, the culture bottles were removed and stared at -20°C. After complete freezing, the culture bottles were thawed rapidly with vigorous shaking to disrupt the cells. The titre of the pooled virus ws found to be 10-4 in kidney cells. 5 percent suspension of the titrated virus was made in Alum Gel to prepare the vaccine. The CPD local virulent virus strain was used for challange purpose. Its titre was calculated and found to be io4.16 in susceptible goats. Sterility of the vaccine waS done on different culture media and safety was carried out in guinea pigs by inoculating the vaccine intramuscularly. The potency of the vaccine W&S done in susceptible goats( Tlilk teeth). The goats which were vaccinated in 10 ml and I ml dosage subcutaneously, and challanged after 14 days of vaccination with I ml of challange dose of 100 RD50 and 1000 RD50, proved to be immune by withstanding the above challange dose. This cell culture CPD vaccine is being used in the field with encouraging results.
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240.
Studies Of The Carriers Of Pasteurella Multocida
by Syed Shabir Ahmad Shah | Prof. Muhammad yousaf Vaid | Mr. Muhammad | Mr. Muhammad Akram Muneer | Faculty of Veterinary Sciences.
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Publisher: 1979Dissertation note: Pasteurellosis is an infectious disease of Livestock having world wide occurrence. To find out the incidence of healthy carriers of Pasteurella multocida in cattle buffaloesa, research project was undertaken at College of Veterinary Sciences, Lahore.
For this purpose 330 (nasopharyngeal) swabs of clinic shy normal cattle and buffaloes were collected from various sources from Lahore.
Inoculations swabs were made on various media like blood agar, tryptore agar, tryptore broth, mutrient agar, nutrient broth, etc. The culture media wee incubated both aerobically and at a temperature of 37oC.
The biochemical characteristics of Pasteurella multocida isolated were studied, smears from growth, were prepared, stained with Gram's method and examined.
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241.
Epizootiology Of Newcastle Disease In Free Flying Birds Of Pakistan
by Muhammad Arshad | Dr. Muhammad Ajmal | Mr. Mubbasher | Mr. Muhammad Naeem | Faculty of Veterinary Sciences.
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; Literary form:
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Publisher: 1984Dissertation note: Epizoàtiology of Newcastle disease was studied in 105 birds each of three species of free flying birds i.e. pigeon, starling and sparrow. Incidence of Newcastle disease was measured on the basis of Haemagglutination Inhibiting antibodies against Newcastle disease virus present in the serum of each bird. It was observed that 42.86% of pigeon, 25.71% of sparrows and 21.90% of starlings were positive for Newcastle disease. An attempt was also made to isolate the virus from faecal material and pooled organs (lung,liver and spleen) of each bird in developing chick embryo. Three strains of Newcastle disease virus, 2 from pizeon and one from starling were isolated. One pigeon having virus in its organs was also found excreting Newcastle disease virus in its faeces. No virus could be isolated from sparrows. The pathogenicity of isolates was studied in day-old chicks by intracerebral inoculation and it was observed that all of the three isolates were moderately pathogenic Though non significant, the percentage of positive cases on the basis of Haemagglutination Inhibition test in male birds was found to be higher than female birds of the same species
From the tudy it was evident that pigeon could play an important role in the transmission of Newcastle disease. The other two species were also susceptible to this virus and could be source of transmission. It recommended that these as well as other species of the ree flying birds in a large number from much wider areas may be studied. It is further recommended that birds found dead in the field and pet bird markets may be investigated for virus isolation.
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242.
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.
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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.
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243.
Staphylococcal Coagglutination Test For Rapid Detection Of Foot And Moth Disease Virus
by Baitullah Khan | Dr. Atif Hanif | Prof. Dr. Masood Rabbani | Faculty of Veterinary Sciences.
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Publisher: 2010Dissertation note: Foot and mouth disease is a highly contagious, viral disease of cloven hoofed animals and causes high economic losses. Rapid detection of FMD is necessary to control the disease from spreading. Although several reliable tests like ELISA, CFT' and PCR already exit but none of them applicable in field conditions. The aim of this study was to optimize rapid, economical and sensitive test for the detection of FMD. For this purpose rabbits were used to raise immune sera against FMD virus with one uninnoculated control. Immune sera collected from these rabbits at different time interval and presence of antibody was determined by using AGPT. Immune sera were then conjugated with satbalized and inactivated staphylococcus aureus cell using different dilutions. Cell wall of S. aureus contains Protein A which naturally binds with Fc portion of IgG leaving the Fab portion to interact with antigen. In presence of homologues antigen causing agglutination was seen with nacked eyes. Light blue background was found best while observing results.
The coagglutination test was applied on FMD known antigen. Clear agglutination on slide was observed by mixing equal quantity of COAT reagent and its respective antigen. Total 40 vesicular fluid samples from FMD infected animals were tested with COAT, in which 38 yielded positive results and the remaining two yielded negative results. COAT reagents were also tested against PPR virus depicting negative results. COAT was found specific for FMD antigens. This test is quick and generates results within five minutes.
This reagents of CAOT also applied on two fold dilution of vesicular fluid from FMD infected animal and positive result were observed up to 1:32 dilution. This test is sensitive, specific, economical and rapid for detection of FMD. This test was successfully used for detection of FMD in filed.
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244.
Identification And Genotyping Of Vp1 Genses Of Fmd Viruses
by Atia Bukhari | Prof. Dr. Irshad Hussain | Prof. Dr. Khushi Muhammad.
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Publisher: 2009Dissertation note: Within two decades after its first report in 1954 from Pakistan, Foot and mouth disease has become endemic in the country and poses a serious threat to large as well as small ruminant population. Foot and Mouth Disease (FMD) is prevailing in cattle and buffaloes and is caused by either 0, A, Asia-i serotype of the FMD virus in Pakistan. The present study was undertaken to study the mutation rate of FMD virus and also molecular typing of the strains prevalent in Pakistan was done.
A total of 60 samples from buffalo and cattle were collected from five districts of Punjab including Lahore, Faisalabad, Sialkot, Okara and Sheikhupura. Soon after extraction of their RNA, all of them were reverse transcribed and then subjected to amplification by using different sets of the primers including universal as well as serotype specific primers. Then their VPI portions were amplified by using VP1 specific primers. Among 60 samples, 48 were positive with universal primers. Other 12 samples were not amplified with these primers hence not processed.
Among 48 FMD positive samples, 24 were positive with serotype 0 specific primers, 16 with serotype Asia-i and remaining 8 were positive with serotype A specific primers. After their amplification, the amplicons were run on the gel. These amplicons were extracted by using DNA extraction kit. After their purification, they were sent to Macrogen® (Seopl, Korea) and Centre of Excellence for Molecplar Biology, Pakistan (CEMB) for sequencing. Each amplicon was sequenced thrice and the consensus sequence was established eliminating sequencing errors.
Sequence identity and multiple sequence alignment of molecular sequences (nucleotide and amino acids) were performed with Clustal W algorithm (Thompson et al., 1994). Neighbour joining trees were constructed by using MEGA version 4.0 (Kumar et al., 2004). Nucleotide distance matrices were computed by Kimura two parameter algorithm based on the total nucleotide substitutions and evolutionary trees for VP1 genes were constructed.
For FMDV serotype '0' phylogenetic analysis, 14 VPI sequences from various field isolates were compared with some previously published Pakistani FMD 0 type VP1 specific sequences available with GeneBank and some recently published VP1 sequences reported by countries bordering with Pakistan including India, Iran and Afghanistan Similarly, 12 VP 1 sequences of FMDV serotype Asia-I isolates of this study were compared with previously published sequences and their phylogenetic relationship was established. However, the sequencing results of serotype A were inconclusive and were not included for phylogenetic analysis. Three sequences of three locally available FMD vaccines were also studied and compared with the outbreak strains.
Polymerase chain reaction was optimized with respect to MgCI2, buffer pH, annealing temperature, primer concentration, template concentration, and Taq polymerase. A concentration of 2.5 mM of MgCl2 resulted in the best amplification of the target sequences (Figure 1). The buffer with pH 8.8 yielded the best results (Figure 2) Although, the suggested annealing temperatures for various primers (of various serotypes) ranged from 48 °C to 63 °C, however, a temperature of 56 °C was found to be the best with all sets of primers (Figure 3). The best intensity DNA bands were observed with 0.3 pM concentration of the primers (Figure 4). Moreover, the best cDNA template concentration giving optimum amplification was found to be 3.0 p1 per reaction (Figure 5). Lastly, a concentration of 0.5 U of Taq polymerase was not sufficient for amplification of cDNAs, however, 1.0 U of enzyme was found to yield better amplification (Figure 6).
VP 1 DNA sequences of six previously published Pakistani FMD serotype 0 strains were analyzed phylogenetically with VP 1 DNA sequences of 14 isolates of the study. Serotype 0 isolates of this study distributed themselves into two distinct clusters (Figure 19). First cluster comprised of Sheikhupura 1 and 2, Muridkey 1, Raiwind 1, Nankana 1, Gujranwala 1 and Gujrat I isolates (Figures 19 and 20), whereas the second cluster included Depalpur 1, Sahiwal 1, Okara I, Multan 1, Toba 1, Faisalabad I and Pattoki 1 isolates (Figures 19 and 21). The first cluster was found to be associated with previously published Pakistani isolates of 2006 mostly. However, it also showed association with Afghanistan's isolates of 2004 (Figure 20). The second cluster seemed to be mostly related to previously published Pakistani isolates of 2003 (Figure 21). The overall grouping of the 14 sequences, when compared with each other, depicted a three clustered phylogram (Figure 22). Serotype 0 isolates from Depalpur, Sahiwal, Okara, Multan, Pattoki, Toba Tek Singh and Faisalabad grouped together into a clan and had more than 85% sequence similarity with each other. The second cluster consisted of isolates of Sheikhupura, Nankana, Raiwind and Muridkey. These sequences had more than 86% similarity with each other. The third cluster consisted of only two isolates which were 100 % similar to each other. However the third cluster had only 74 % sequence similarity to cluster I and 73 % sequence similarity when compared with cluster 2.
When the phylogenetic relationships with previously reported isolates of Asia 1 was evaluated, FMD Asia I isolates of this study were found to be scattered into two distinct groups (Figure 16). Group one consisted of isolates of Lodhran, Toba and Hafizabad that were more closely related to Indian isolates sharing more than 98% identity with each other and more than 94 % sequence identity with isolates of Indian 2001 to 2004 (Table 5 and Figures 16 and 17). However, they shared more than 86% sequence similarity with Pakistani isolates of 2002-2005 (Table 5). Group two comprised of isolates of kasur, Lahore, Pakpattan, Okara, Faisalabad, Jhang, Rahim Yar Khan, Bahawalpur and multan alongwith vaccine A and B (Figure 16). The isolates of group 2 were found to be closely associated with previously published isolates of Pakistani and Afghani origin of year 2003 and 2004 (Figures 16 and 18). Collectively, they shared an overall 70% sequence identity with each other. However, isolates of Bahawalpur, Rahim Yar Khan and Multan shared more than 98% similarity with each other, a measurement of close relationship denoting a likely common origin as one clan or dade. Similarly, isolates of Pakpatan, Faisalabad, Okara, Kasur, and Lahore shared 88% sequence identity with each other and qualified as one clade.
Although, overall amino acid sequence similarity of our isolates was not strikingly different from that of the published isolates, however, amino acid substitutions with dissimilar properties were found with a scattered pattern of distribution. For example, 15th amino acid residue which is hydrophilic in the previously published isolates had a substitution with a hydrophobic amino acid residue in our three isolates namely Sheikhupura 2, Muridkey I and Raiwind I (Figure 25). Similarly, 14th amino acid residue which is hydrophobic in nature was found to be replaced with a hydrophilic one in our last five isolates. Amino acid residue number 13 (Figure 25) had a substitution with a hydrophobic residue in some of our isolates etc. etc. It is interesting to note that such substitutions with amino acids having dissimilar properties have also been found, albeit at lower rate, in previously published sequences by many researchers (Figure 25).
A comparison of the deduced amino acid sequences in the critical VP I region of FMD serotype Asia I revealed that most of this study isolates shared very high homology with sequences of Vaccine A. However, the sequences of isolates of Lodhran, Hafizabad and Toba did not match much with that of either vaccines, A or B (Figure 23). Sequences of Vaccine A had a "K" which seemed to be replaced by a "T" in the sequences of most of the isolates. Considering the properties of various amino acids, this change does not signify a major shift in the three dimensional picture of the protein as K is a lysine, a positively charged amino acid, whereas a T is threonine, a hydrophilic amino acid in nature. Next substitution in most of the isolates is a "P" for "A" in comparison to the vaccines. Again, it is not a significant change as both P and A share the same property, hydorphobicity. Similarly a K with an R can be substituted without much change in the overall shape of the protein molecule. Next amino acid substitution is a leucine instead of methionine. Again both are hydrophobic in nature; hence their impact on the overall picture is minute, if at all. However, glycine and arginine are two very different amino acids; the former is a hydrophobic amino acid whereas the latter is positively charged one. Such amino acid substitutions may have the potential to make a major impact in terms of the epitopic differences in the capsids of vaccinal and field viruses. A comparison of the deduced amino acids of FMD serotype 0 isolates also exhibited such changes with the vaccinal virus (Figure 24).
Of the three hyper immune sera raised against three different vaccines in rabbits, only one vaccine induced a measureable immune response yielding good precipitation line against various FMD virus antigens.
In summary, RT-PCR for diagnosis of serotypes A, 0 and Asia 1 of FMDV was optimized and could be used for prompt and precise diagnosis of FMD in the country. Although, RT-PCR data pertains to bovines in the current project, but PCR optimization parameters are equally applicable to FMDV infections in other FMD susceptible animal species such as sheep and goat. The combination of PCR and sequencing of the VP1 gene to detect and analyze FMDV in disease outbreaks is fast (less than 6 hours for PCR and about 24 hours for sequencing), and it can give an accurate immunologic characterization of the virus, thus providing a rational basis for choice of vaccine. In fact, the molecular epidemiology of field isolates is a powerful tool to monitor the circulation of viruses (Saiz et al., 1993).
Secondly, various isolates of serotypes 0 and Asia 1 were sequenced along with some vaccinal strains. Sequence similarity tree analysis indicated that most of our isolates were closely related to previously reported Pakistani isolates and to those of neighboring countries such as India, Afghanistan and Iran. Additionally, amino acid sequence similarity data of major immunogenic site that forms 13G-13H loop in FMDV serotypes revealed that serotype Asia 1 vaccinal strain and Asia 1 isolates of this study possessed high degree of similarity suggesting a likely host immune response against the vaccine that may afford some protection against most field isolates of serotype Asia 1 type. Lastly, of three vaccines tested, only one was found to afford protection against field isolates of FMDV suggesting more work on vaccine issue in the country.
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245.
Food Borne Bacterial Contamination Of Retail Poultry Meat At Chicken Sale Outlets In Lahore City
by Anwar Ullah | Dr. Aftab Ahmad Anjum | Dr. M. Younus | Dr. Tahir yaqoob.
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Publisher: 2010Dissertation note: Poultry is an important sub sector of live stock and its share in GDP is 2.0 percent and it is playing an important role in improving the income of rural and urban population of Pakistan. Food poisoning is becoming a serious health problem to human being. The organisms belonging to the Genera Salmonella and Campylobacter are considered to be the most important pathogens. The present research is performed for isolation and enumeration of bacterial pathogens from the family Enterobacteriaceae and Genus Staphylococci.
A total of 200 samples each of liver, kidneys, breast muscle and intestine were collected from four different roads of Lahore city. Each sample was weighed 5gm and was rinsed in 10 ml sterile phosphate buffer saline (PBS) for collection of Sample rinses. Serial 10-fold dilutions of sample rinses (SR) were subsequently prepared in sterile phosphate buffer saline (PBS) for standard plate count (SPC). Each dilution of sample rinsed were inoculated to selective agar plates for enumeration and isolation S. enteritidis, C. jejuni, E. coli and S. aureus.
Numbers of selective agar plates contaminated with E. coli were 75.5%, having the highest value and S. aureus were isolated from the lowest number of plates (50%). C. jejuni were isolated from 119 (59.5%) out of 200 samples and Salmonella enteritidis contamination rate were 71% (142 out of 200 samples). Over all 200 samples each of liver, kidneys, breast muscles and intestine 158(79%) out of 200 were contaminated with food borne bacteria. Kidney samples were less contaminated having contamination rate of 42.5% (85 out of 200 samples). Intestine and liver samples were contaminated with a medium range of 66.5% (133) and 68% (136) respectively.
Intestinal samples had highest range of colony forming units (CFU) per gram of C. jejuni and kidney samples had the lowest CFU per gram with an average CFU per gram of 1.2 x106 (<log 6.09>) and 1.0 xi (<log 4.02>)respectively. Liver samples had an average CFU per gram of 6.3 xl ü (<log 4.80>) and breast muscle had 1.4 x ] (<log 5.15>)CFU per gram of sample(Table 02). Liver samples had highest CFU per gram of S. enteritidis and kidney samples had the lowest value with an average CFU per gram of 1.1 x106 (<log 6.05>) and 9.8 xi0 (<log 3.99>) respectively. Intestinal samples had an average 9.7 xiO5 (<log 5.98>) and breast muscle had 1.1 xi (<log 5.04>) CFU per gram of sample (Table 03). Intestinal samples had highest CFU per gram of E. coli and kidney samples had the lowest value with an average CFU per gram of 7.7 x l0 (<log 7.88>) and 1 .5 x (<log 4.19>) respectively. Liver samples had an average 8.7 x 0 (<log 5.94>) and breast muscle had 1.8 x 106 (<log 6.25>) CFU per gram of sample (Table 04). The breast muscles had the highest CFU per gram of S. aureus and kidney samples had the lowest value with an average CFU per gram of 1.1>i0 (<log 5.04>) and l.5x 10 (<log 4.17>) respectively. Liver samples had an average of 1.1 xl (<log 5.05>) and intestinal samples had 2.5x10 (<log 4.40>) CFU per gram of samples.
The samples were sub cultured for isolation and identification of food borne pathogens like Salmonella enteritidis, Campylobacter jejuni, Escherichia coil and Staphylococcus aureus species by following the standard protocols of Bergey's Manual of Determinative Bacteriology 9th Edition.
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246.
Development And Optimization Of Multiplex Pcr For The Identification Of A, O And Asia 1 Strains Of FMDV In Pakistan
by Muhammad Ikram | Dr. Atif Hanif | Dr. Imran Najeeb | Prof. Dr.
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Publisher: 2010Dissertation note: Foot and mouth disease (FMD) is highly infectious disease of cattle, buffalo, sheep and goats. It is caused by genus Aphthovirus of Picomaviradae family. FMDV is RNA virus having seven serotypes A, 0, C, Asia I, SAT1, SAT2 and SAT3.
Foot and mouth disease is endemic in Pakistan and causes high economic losses to livestock industry. So priority is to develop quick and efficient methods for detection of FMDV and to limit the spread of disease outbreak. Although CFT, VNT and ELISA are already being used for the diagnosis of FMDV in Pakistan but these diagnostic techniques are time consuming and their specificity and sensitivity is low. Multiplex PCR for the identification of FMDV is very much sensitive and specific, can be done with in three hours after the receipt of samples.
Present study has been designed to optimize multiplex RT-PCR for rapid detection of FMD virus. RNA was extracted from virus stock obtained from QOL, UVAS Lahore and from field samples. After RNA extraction the samples were subjected to synthesize cDNA by the use of Reverse Transcriptase enzyme. After cDNA synthesis PCR reaction was carried out. The amplified products were resolved on 1.5% Agarose Gel. A multiplex RT-PCR strategy was optimized and developed for the detection of virus serotypes A, 0 and Asia l.
Restulst of this study helped to develop an efficient and economical method for rapid detection of FMD virus and also helpful in differential diagnosis from other vesicular diseases.
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247.
Microbial Evaluation Of Raw Meat At Abattoirs And Retail Outlests (Lahore)
by Abid Sarwar | Prof. Dr. Mansur ud Din Ahmad | Dr. Imran Najeeb | Prof. Dr. Azhar.
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Publisher: 2010Dissertation note: The objective of this study was to evaluate the microbial quality of meat. The present study was planed to determine the aerobic plate count on meat obtained from the abattoirs and local market. A total of 90 meat samples that were collected for determining the microbiological quality of meat. Half of the meat samples (n=45) were collected from various abattoirs and half of the meat samples (n=45) were collected from retail outlets in Lahore City to get an idea of contamination from slaughtering point to retail outlets.
These samples were processed for Aerobic plate counts, E.coli, S.aureus and Salmonella counts. Overall, this study revealed that the level of contamination on meat carcasses was higher in retail meat shops compared to the abattoir. However, the microbial contamination in the abattoir were high if we compare these results to the reports from developing countries like India, Iran and Bangladesh.
Bacterial isolates identified and counted from this study were Staphylococcus aureus (44) out of 90 samples was the most abundant as 48.88%, followed by E. coli (43) 47.77% and Salmonella (26) 28.88%.
Statistical analysis revealed that analysis of variance between various abattoir and the retail meat shops for E.coli, Salmonella and S.aureus showed significant differences with some exceptions. E.coli counts were significantly higher (P < 0.05) in the meat shops and abattoirs. For E.coli most of the data were significant at 5% level (P < 0.05) with some exception in case of beef and goat samples taken from abattoirs which were non significant because of the unhygienic environments. Analysis of variance for Salmonella between various abattoir and the retail outlets were significant at 5% level (P < 0.05). For S.aureus between various abattoir and the retail outlets showed non significant at 5% level (P > 0.05) with some exceptions in case of beef abattoir and goat retail outlet samples taken which were significant at 5% level (P < 0.05).
The higher incidence of microbial load in fresh meat obtained in this study might be attributed to unhygienic and improper handling of animals during slaughter, dressing, evisceration, transportation and unhygienic environments at the retail shops. The usual practice of washing the carcass with the same water in which intestines and offal had been washed was considered as one of the predominant reasons for increased microbial counts of the carcasses. A complete ignorance on the part of the meat handlers/ butchers in hygienic handling of carcasses during slaughter and retailing processes might be the main factors for producing meat with high microbial load.
Levels of microbial contamination in Pakistani abattoirs and traditional retail meat shops reflect the hygiene status of meat production in the developing world. Education of the meat retailers' community which runs the traditional meat shops, in terms of the importance of hygienic and sanitary precautions would go a long way towards providing wholesome and safe meat to the consumers.
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248.
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.
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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).
249.
Characterization Of Indigenious Species Of Mycotoxins Producing Aspergilli
by Gull Naz | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.
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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.
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250.
Detection Of Hazardous Organism In Raw And Pasteurized Milk With Particular Reference To 3Enterobacteriaceae
by Ayesha | Prof. Dr. Mansur ud Din Ahmad | Dr. Aftab Ahmad Anjum | Prof. Dr.
Material type: ; Format:
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Publisher: 2010Dissertation note: The present study was carried out to detect the hazardous organisms in raw milk from public health point of view. In total one hundred twenty (120) milk samples were collected from milk retail shops in and around Lahore. Out of these 120, one hundred samples were of raw milk and rests of the twenty samples were of pasteurized milk. Their microbiological quality was studied by performing standard plate count (SPC), coliform count and identification of hazardous bacteria belonging to the family Enterobacteriaceae. The micro flora of milk was also studied for the prevalence of multiple drug resistant (MDR) bacteria.
Milk supplied in Lahore city was found to have poor microbiological quality. Bacterial load was determined by SPC and coliform count. The standard plate count (S.P.C) of the raw milk ranged from 4.2x106 to 7.7xl07 c.f.u/ml. The coliform counts ranged from 3.4x 104 c.f.u /ml to 6.9x105 /ml. A total of 81 isolates were identified from raw milk samples. These included Yersinia (3 strains), Klebsiella (16 strains), Escherichia coli (14 strains), Enterobacter (11 strains), Shigella (3 strains), Salmonella (19 strains) and' Proteus (15 strains).The standard plate count for pasteurized milk ranged from 1.45x104 c.f.u/ml to 3.8x 105 c.f.u/ml. The minimum and maximum coliform count was 7.2x102 to 8.4xl03 c.f.u/ml respectively for pasteurized. All samples were outside the international standard for coliform bacteria. A total of 13 isolates were identified from pasteurized milk samples. These included Yersinia (2 strains), Klebsiella (1 strains), Escherichia coli (6 strains), Enterobacter (2 strains), Shigella (1 strains) and Proteus (1 strains).
All the isolates showed multiple drug resistance to various commonly used antibiotics in veterinary practices. Escherichia coli were resistant to all antibiotics used except Gentamicin (10µg). Enterobacter was sensitive to all the antibiotics used except to Ampicillin (10µg). Shigella was sensitive to Gentamicin (10µg), Kanamycin (30µg), Choloramphenicol( 25µg), but showed resistance to Ampicillin (10µg), Oxytetracycline ( 25µg), Streptomycin (10 µg), Pencillin (10 µg) and Tribrissin (25µg)., Salmonella was resistanct to Ampicillin (10µg), Oxytetracycline ( 25µg), Streptomycin (10 µg), Pencillin (10 µg) and Tribrissin (25µg). But sensitive to Gentamicin (10µg). .All the isolates showed greatest resistance to Penicillin (10 ug.) whereas, most of the isolates were sensitive to Gentamycin, Kanamycin and Chloramphenicol.
Finally, it is recommended that the members of the public should always boil raw milk before consumption because of their microbial content. Therefore, it is highly recommended that hygienic practices and regulations, such as on-site pasteurization and implementation of HACCP following established standards, should be introduced to facilitate the production of raw milk of high quality and safety.
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