251.
Physicochemical Factors Affecting Infectivity Of Pesti Des Petits Ruminants Virus
by Kinza Khan | Prof. Dr. Khushi Muhammad | Dr. Jawad Nazir | Dr. Mutti-ur-Rehma.
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Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1636,T] (1).
252.
Comparative Efficacy Of Hand Sanitizers And Liquid Soaps Against Commonly Encountered Microbes On The Experimentally Contaminated Palm Surfaces
by Taiba Tahir | Dr.Jawad Nazir | Prof Dr | Prof Dr. Khushi Muhammad.
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Publisher: 2013Dissertation note: Hand hygiene plays a key role in the prevention, control and reduction of many communicable infections as contaminated hands are the major source of transmission for microbes. Three categories of hand hygiene products; hand sanitizers (Safeguard, Dettol, and Cool & Cool hand sanitizers), antibacterial soaps (Safeguard, Dettol, and Lifebuoy liquid soaps) and plain soaps (Lux, Capri, and Pears liquid soaps) were evaluated against five bacterial cultures (E. coli, K. pneumonia, Salmonella. spp, S. aureus, P. aeruginosa) for their antibacterial activity through in vivo and in vitro techniques. In vivo testing was performed through palmar surface contamination techniques. Palm surfaces of volunteers' hands were artificially contaminated followed by recovery of the bacteria through glove juice method both before and after the application of product for 30 seconds. Each of the experiment repeated thrice and means log reduction (MLR) in the bacterial count after the application of each product was calculated. In vitro efficacy of hand hygiene products was carried out through calculation of minimum inhibitory concentration (MIC) and phenol coefficient values.
MLR values of the sanitizers were ranged from 2.0 - 5.5 log10 CFU/ml, while that of antibacterial and plain soaps were 3.0 - 4.1 and 3.0 - 4.6 log10 CFU/ml. MIC values for the sanitizers, antibacterial, and plain soaps were ranged from 1:10 - 1:40, 1:6 - 1:20, and 1:2 - 1:8 against all of the 5 bacteria. Hand sanitizers were proved to be superior to medicated and plain soaps during in vivo and in vitro testing. Both of the antibacterial and plain soaps were equally effective in reducing bacterial load on the contaminated hands because during hand washing procedure mechanical removal of contaminants through surfactant activity of soaps is mostly responsible for the removal of bacteria. While a relatively higher MIC values of the antibacterial soaps were attributed to the presence of certain antibacterial agents in them. It was not possible to calculate the phenol coefficient values for any of the hand hygiene product because even least dilutions (1:2) of the products did not stop the bacterial growth. Present study emphasizes the suitability of using hand sanitizers in health care centers as well as in routine life. Because of comparable efficacy of medicated and plain soaps, excessive use of antibacterial soaps should be avoided due to risk of developing antibiotic resistance.
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253.
Isolation And Characterization Of Auxin Producing Bacterial Strains From Plant Rhizosphere
by Kanwal Aziz | Dr. Jawad Nazir | Dr | Prof. Dr. Tahir Yaqub.
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Publisher: 2013Dissertation note: Auxins are a class of plant hormones or plant growth substances. Auxins have a cardinal role in the regulation of many growth and developmental processes in the plant. Soil samples were serially diluted and screened for auxin production by Salkowski method. Isolates that have the ability to produce auxin were identified by culture characters, morphology, and bio-chemical profile. From 150 isolates, 04 bacteria were selected (AUX-36, AUX-53, AUX-137, and AUX-142). The bacteria were identified by following the flowcharts described in “Berges Mannual of Determinative Bacteriology”, 9th addition. These isolates were identified as Bacillus megaterium, Escherichia coli, Klebsiella pneumonia and Bacillus marinus respectively. Next, different physical and chemical parameters for growth of bacteria and auxin biosynthesis were optimized. For the optimization of bacterial growth OD values of the culture broth (at wavelength 600nm) was taken by spectrophotometer. To estimate the amount of auxin (ìg/ml) produced in the culture broth; a standard curve (concentration of auxin ìg/ml at x-axis and OD value at y-axis) was prepared by using commercially available auxin.
The optimum conditions for growth and auxin production by AUX-36 was found to be pH 7, 0.98% osmotic pressure at 37 °C after 72 hours of incubation. If the medium is supplemented with 0.1 and 1.0% glucose, sucrose and peptone then it increased the bacterial growth which ultimately increased the auxin concentration in the broth medium. The growth and auxin
Production by AUX-53 was 0.98% NaCl concentration at 37 °C after 72 hours of incubation. The optimum pH was found to be 7 but it showed good growth at acidic as well as alkaline pH. The addition of glucose and sucrose in the growth medium increased the growth as well as auxin production. The optimum conditions for the growth of AUX-137 were as follows: pH=7, 0.98% osmotic pressure, temperature 37 °C. However the isolate had good growth at 28 °C and 2% NaCl concentration as well. The bacterial cell density and auxin increased with incubation time up to 72 hours. The isolate produced highest concentration of auxin under the same conditions. Similarly, the cell density and auxin increased with the increasing concentration of glucose in the growth medium. Sucrose increased the auxin only in the culture filtrate. While the bacteria AUX-142 showed highest growth as well as auxin production at 42 °C after 72 hours of incubation. The optimum pH and osmotic pressure was found to be 7 and 2% respectively. The cell density and concentration of auxin increased with the increasing concentration of peptone in the growth medium. Addition of tryptophan (1-2%) increased the auxin concentration in the culture supernatant of all isolates.
Next, the seed germination test and plant pot experiment were performed of selected isolates to observe the effect of bacterial inoculation on wheat plants. In seed germination test treatment of seeds with AUX-36, AUX-53 and AUX-142 significantly increased the root length and number of root hairs as compared to non-treated seeds. In plant pot experiment comparison of various growth parameters of inoculated plants with non-inoculated plants revealed the improvement in plant growth by bacterial inoculation.
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254.
Isolation And Characterization Of Phytase Producing Microrganism From Soil
by Ghazal Aziz | Dr. Aftab Ahmad Anjum | Prof. Dr | Prof. Dr. Tahir Yaqub.
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Publisher: 2013Dissertation note: Phytase is an enzyme of great importance because it is added as a biofertilizer to soil and added in animal feed to increase the uptake of inorganic phosphorous. Phytase production is the property of plant growth promoting rhizobacteria (PGPR) that harbor in rhizosphere part of the soil. These phytase producing bacteria can be utilized as biofertilizers as and can increase the soil fertility and crop production.
Soil samples were collected and screened for the production of phytase (an extracellular) enzyme on phytase screening media (PSM). Six bacterial isolates (PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30) showed distinguished clear zones (> 6mm) on PSM. Isolates were identified as Lactobacillus casei PHY02, Enterobactor intermedius PHY03, Bacillus badius PHY06, Escherichia coli PHY07, Shigella sonnei PHY12, and Klebsiella pneumonia PHY30.
Effect of physical parameters (temperatures, pH and osmotic pressure) on growth and enzyme production by selected isolates was determined. Optimum growth and production of phytae by PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30 (27, 9, 19, 40, 32, and 19 IU, respectively) was at 37°C. PHY07 showed highest enzyme production, followed by PHY30 and PHY02. Isolate PHY06 showed similar growth and enzyme activity at 37°C and 42°C but it was significantly reduced at low temperature. Effect of pH on phytase production on selected isolates indicates that all isolates produces maximum amount of phytase at pH 6.5. At pH 6.5 enzyme units released by PHY02, PHY03, PHY06, PHY07,
PHY12, and PHY30, were 26, 15, 19, 41, 19, and 32 IU, respectively. Production of enzyme decreased with the increase in osmotic pressure. PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30 showed optimum enzyme production (27, 15, 17, 41, 18, and 32 IU, respectively) at 1 % NaCl in PSM (Figure 1C).
Effects of carbon source on both growth and phytase production of isolates showed that PHY03, PHY06, PHY07, PHY12 had significantly higher (P<0.05) cell densities and enzyme production in glucose, while PHY02 and PHY30 had higher enzyme activity at 0.3% lactose.
Nitrogen source in growing media also effects the growth and production of enzyme. PHY02 and PHY12 had better growth and production at 0.1% peptone, while PHY07 and PHY30 had significantly higher phytase level in media modified with peptone but at higher concentration (0.3%). Addition of tryptone in growth medium significantly enhanced the growth and enzyme production by PHY03, and PHY06.
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255.
Toxinotyping And Antimicrobial Susceptibility Of Enterotoxigenic Clostridium Perfringens Isolates From Muttion, Beef and Poultry Meat
by Madiha Khan | Dr. Jawad Nazir | Dr. Aftab Ahmad Anjum | Prof. Dr.
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Publisher: 2013Dissertation note: A total of 300 meat samples including chicken, mutton, and beef (100 each) collected from local butcher shops as well as large meat outlets and grocery stores situated in various localities of Lahore were analyzed to determine the level of C. perfringens contamination. The samples were enriched in Fluid Thioglycollate Medium (FTM), purified on Tryptose Sulfite Cycloserine (TSC) agar that is highly selective media for C. perfringens and were identified by their culture characters, morphology and biochemical profile. C. perfringens was successfully isolated from 12 out of 300 samples with an overall positivity ratio of 4 %. A relatively higher percent prevalence of the C. perfringens was found in meat from local butcher shops (6.66 %) in comparison to the ones collected from the larger meat outlets (1.33 %) where meat is supplied under cold chain management system. Within each meat type a total of 6, 5, and 1 of the samples from chicken, mutton, and beef meat, respectively were found positive for the presence of C. perfringens.
Toxinotyping of the positive isolates was performed using commercially available alpha, beta, and epsilon toxins detection ELISA kits. Out of 12 confirmed isolates of C. perfringens only six were found positive for the production of various toxins. Three of the isolates produced alpha toxin and were grouped as type A, one of the isolate produced alpha, beta and epsilon toxin therefore confirmed as type B, one of the isolates produced alpha and beta toxin so belong to type C whereas one of the isolate produced alpha and epsilon toxin so it was grouped as type D while six of the isolates did not produce any toxin.
The toxin producing isolates were subjected to antibiotic susceptibility testing against 13 antibiotics commonly employed to treat the foodborne infections. It was observed that most of the antibiotics were effective against C. perfringens exhibiting a wider zone of inhibition around the antibiotic discs. All the six isolates were susceptible to the chloramphenicol, ciprofloxacin, metronidazole, and ceftriaxone. Five out of six isolates were susceptible whereas one of the isolate was classified as intermediate against tetracycline, lincomycin, and cefotaxime. Five isolates were sensitive and one was resistant to erythromycin. Four isolates were susceptible to penicillin and one each was intermediate and resistant to the antibiotic. All of the other drugs were relatively less effective with a least activity of amoxicillin against the isolates.
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256.
Isolation And Characterization Of Multidrug Resistant E. Coli From Urinary Tract Infections In A Tertiary Care
by Sumera Sabir | Dr. Aftab Ahmad Anjum | Dr | Dr. Muhammad Asad Ali.
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Publisher: 2013Dissertation note: Bacterial etiology of urinary tract infections (UTIs) admitted in or visiting a tertiary care hospital in Lahore, Pakistan was determined by conventional biochemical profile. Multiple drug resistance (MDR) of Escherichia coli, the most prevalent bacteria, was checked. Overall bacterial prevalence recorded was 80.4 percent, being highest in patients of intensive care unit (93%) followed by urology ward (87%), north surgical ward (85%), east medical ward (70%) and OPD (67%). Infection rate was higher in female (87.5%) than male (71.3%) and almost same in pregnant (86%)/non-pregnant (88%) female patients. Highest percent UTIs observed were in patients of 51-75 years of age. Percent infection recorded in catheterized patients (70.8%) was lower than non-catheterized (83%) and little higher in Diabetics (82%).
Out of biochemically identified bacterial isolates (n=402), highest number was of E. coli 321 (80%) followed by Staphylococcus aureus 38 (9.4%), Proteus species 22 (5.4%) and Pseudomonas species 21 (5.2%). Almost same pattern of isolation was observed among patients of different wards. On statistical analysis significantly higher number of E. coli was observed among isolates from patients of five wards included in study plan. Out of bacterial isolates from male (n=157) and female (n=245) patients highest prevalence was of E. coli (79% and 80%). Out of total bacterial isolates from female patients (n=245), number of was E. coli at the highest rank 90 (79.6%), in pregnant.
Among different age groups highest prevalence was of E. coli and lowest of Pseudomonas species. Out of 120 tested urine samples collected from catheterized patients bacterial growth was observed in 85. On bacterial identification by conventional biochemical characterization highest prevalence was of E. coli (56.4%). Out of pure bacterial cultures (n=70) from Diabetic patients highest number identified was of E. coli 54 (77.1%) followed by Staphylococcus aureus 8 (11.4%), Proteus 2 (2.8%) and 6 (8.57%) were Pseudomonas species.
According to Antibiotic sensitivity testing results E. coli showed highest resistance to penicillin/amoxicillin (100%) followed by cefotaxime (89.7%), ceftazidime (73.8%), Cephradin (73.8%), tetracycline (69.4%), doxycycline (66.6%), augmentin (62.6%), gentamycin (59.8%), cefuroxime (58.2%), ciprofloxacin (54.2%), Cefaclor (50%), Aztreonam (44.8%), ceftriaxone (43.3%), imipenem (43.3%), streptomycin (30%), kanamycin (19.9%), Tazocin (14%), Amikacin (12.7%) and lowest to norfloxacin (11.2%). Out of 321 E. coli 261 (81%) were declared MDR being resistant to three or more antibiotic classes. Most of the urinary tract infections in human beings are caused by E. coli which show resistance to multiple antibiotics.
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257.
Preparation & Evaluation Of Combined Pre & Fmd Vaccines For Small Ruminants
by Zanira Shakoor | Dr. Jawad Nazir | Prof. Dr. Khushi Mohammad.
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Publisher: 2013Dissertation note: PPR is an acute, febrile and contagious viral disease of small ruminants caused by Morbillivirus. Similarly, FMD is another highly contagious viral disease of cloven hoofed animals that affects more than 33 species of domestic and wild animals. Although disease severity is relatively higher in large ruminants but small ruminants might also play an important role in the epidemiology and transmission of FMD. Mass vaccination is the most effective mean to control viral diseases like PPR and FMD in small ruminants. The present study was designed to evaluate the immune response of goats to various monovalent and bivalent as well as adjuvant and non adjuvant PPRV and FMD "O" virus vaccines. A total of nine groups of animals each comprising 5, were inoculated with various formulations of the vaccines. Serum samples were collected at the beginning and at 1, 2, and 3 months post vaccination. Mean neutralizing antibody titers (MNA) against PPRV and MNA along with complement fixing (CF) antibody titers against FMD "O" in the serum samples were measured to test the efficacy of the vaccines.
Live attenuated wet, gel and oil based PPR vaccines induced 229.2 (±112.79), 253.6(±83.05), and 424 (±182.06) MNA titers in the goats after 3 months of vaccination. Similarly, animals inoculated non-adjuvant, gel based, and oil adjuvant FMD "O" vaccines developed 20(±4.35), 186.2 (±65.39), and 285.8 (±63.80) MNA and 0.4 (±0.894),22.4(±8.76) and 25.6(±8.7) CF antibody titers at 3 months post vaccination (PV). Bivalent (PPRV and FMDV) non adjuvant vaccine induced 249.6(±3.130) and 27.2(±14.53), gel based vaccine induced 306 (±99.82) and 296.2 (±58.21) while the oil based vaccine provoked 417.8 (±141.56) and 286 (±97.13) MNA titres against PPRV and FMD "O" virus respectively, at 3 months PV. Results of present study demonstrate that live attenuated PPRV vaccines can induce better immune response in comparison to killed FMD "O" virus vaccine. Addition of adjuvants is helpful to enhance the immunogenicity of the added antigens. While adjuvant bivalent vaccines containing PPRV and FMD "O" virus can effectively provoke equivalent antibody response against both of the immunogens in the vaccine.
Antibody titee in response to Bivalent vaccine of PPR and FMD seotype"O" containing oil as an adjuvant was superior to gel adjuvant and non adjuvant bivalent vaccines. There was poor immune response of goats to non adjuvant FMD"O" vaccines.
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258.
Isolation Identification & Molecular Based Investigation Of Bovine Rotavirus
by Ambreen Masood | Prof. Dr. Tahir Yaqoob | Dr. Jawad Nazir | Dr. Sehrish Firyal.
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Publisher: 2013Dissertation note: Livestock is an important part of the economy of Pakistan. Calf's diarrhea due to group A bovine rotavirus causes high morbidity and mortality, which results in significant economic losses to livestock. In Pakistan overall prevalence of bovine rotavirus infection is 2.6%. As Pakistan is a developing country, survival of calves is really important to produce milk, meat and hides for propagation of livestock. The aim of current study was to isolate bovine rotavirus from faecal samples of diarrheic calves by antigen capture ELISA and molecular investigations. So, it was helpful to check the prevalence of bovine rotavirus in Lahore district.
This study will be a milestone for better treatment strategies of calf diarrhea problem. It will also pave the way for better vaccine development strategies to cure the disease. A total of 100 diarrheic faecal samples of cattle and buffalo's calves less than three months of age were collected from Lahore district. Rotavirus screening was done by direct sandwich ELISA by using commercial Rotavirus detection kit (Cypress Diagnostics, Belgium). ELISA confirmed 12 samples to be positive for bovine rotavirus. Among 12 positive samples, 7 were found positive in buffalo calf and 5 in cattle calf. After RNA extraction and cDNA synthesis, the PCR was done for amplification of VP4 gene of all ELISA positive bovine rotavirus samples. But only 5 samples (3 buffalo calf samples and 2 cattle calf samples) give desired product of 880 bp of VP4 gene. After sequencing and bioinformatics analysis, phylogenetic tree was constructed. It is evident that Pakistani bovine rotavirus VP4 gene (BRV/QOL/13) has maximum identity of 98% with Indian bovine rotaviruses VP4 gene.
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259.
Production And Evaluation Of Peste Des Petits Ruminants Virus Vaccine
by Muhammad Aness | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani.
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Publisher: 2013Dissertation note: PPR is an acute highly contagious viral disease of small ruminants which is endemic in Pakistan. Present study was aimed to evaluate the freeze dried PPRV vaccine with variable biological titer to induce protective immune response in beetal goats. The comparative immune response of animals to adjuvant and non-adjuvant vaccine and duration of immunity was also studied. Effect of boosting on the humoral immune response of animals as well as shelf life of various vaccines was also evaluated. Each of the vaccines was inoculated in a group of five animals. Serum samples were collected at specified time intervals and antibody levels were detected through cELISA as PI values and neutralization test as MNA titer.
The virus was propagated on the Vero cells. It was estimated that infecting 2 x 107cells with 104.00 TCID50 virus concentrations added to a T-175 cell culture flask at the time of subculture yielded maximum virus titer in the cell culture harvest following three freeze thaw cycles of the contents. The freeze dried vaccine with a biological titer of 105.00 TCID50 per dose provoked maximum antibody titer followed by the ones with a titer of 104.00 or 103.00TCID50 which provoked nearly equivalent protective immune response while the animals inoculated with a vaccine having 102.00 TCID50virus concentrations developed minimum antibody titer. The oil adjuvant PPRV vaccines elicited significantly higher antibody titer in comparison to gel based vaccines but however minimum antibody titers were detectable in response to freeze dried vaccines. Although protective antibody level (? 10 neutralizing antibody units) was detectable in the animals vaccinated with either oil based, gel based or freeze dried vaccine containing biological titer of 104.00 TCID50 but however the extent and duration of immunity was found to be most superior in response to oil based vaccines. It can be concluded that a single shot of either gel or oil based vaccine can provide protection in the vaccinated animals for a minimum of one year duration. Goats receiving a booster dose of the vaccines had a significantly higher antibody tier in comparison to the ones who received single dose of the vaccines. The freeze dried and wet vaccine kept at 4 °C did not show any significant drop in the biological activity of the virus even after 12 months of storage. Immunogenicity of the both adjuvant and non-adjuvant vaccines, as measured through the immune response in the vaccinated animals, also remained unaffected after 12 months of storage at 4 °C.
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260.
Antibody Response Of Buffalo Calves To Oil Based Multivalent (Pasteurella Multocida, Clostridium Chauvoei And FMD Virus "O' "A" and "Asia1") Vaccine
by Muhammad Farooq | Prof. Dr. Khushi Muhammad | Dr. Aftab Ahmad Anjum | Prof. Dr.
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Publisher: 2013Dissertation note: Microbial diseases are one of the constraints for further development of dairy industry as a profitable enterprise. The diseases are causing heavy economic losses to the industry. The diseases such as Foot and Mouth Disease (FMD), Hemorrhagic Septicemia (HS), Black Quarter (BQ),etc., are endemic in Pakistan and perpetuate among the dairy animals. These diseases can be effectively controlled by vaccination.
FMD virus "O", "A" and "Asia 1" were grown on BHK-21 and were inactivated with BEI. Culture of P. multocida and Cl. chauvoeiwere grown on CSY and RCM media, respectively and inactivated with formalin. The vaccine containing 0.2 x 107 units of TCID50of each serotype of FMD virus ("O", "A" and "Asia1"), 2 mg of Pasteurella multocida and 250 Hemolytic units of Clostridium chauvoei per dose were prepared. Oil adjuvanted vaccines of HS, HS + BQ, HS + FMD ("O", "A" and "Asia 1"), BQ, BQ+ FMD ("O", "A" and "Asia 1"), FMD ("O", "A" and "Asia 1") and HS + BQ + FMD ("O", "A" and "Asia 1") were prepared and injected into the buffalo calves in 7 group of 3(n=3) animals each separately at Living Dairies, Chunian. 8th group of three animals was kept as negative control. Antibody response against FMD virus, Cl. chauvoei and P. multocida were measured by CFT, Anti hemolytic Assay and IHA, respectively at day 0, 30, 60 and 90 post vaccinations. Two groups (n=3) of calves vaccinated with whole culture FMD vaccine and NSP free FMD vaccine. Data was analyzed by one way ANOVA procedure and significance was determined by Duncan Multiple Range Test through SPSS version 13.
The vaccine when injected in buffalo calves induced Log22.00±1.00units of anti FMD "O" CFT antibody titer, Log22.22±1.00 units of anti FMD "A" CFT antibody titer, Log22.22±0.84 units of anti FMD "Asia 1" CFT antibody titer; Log22.99±0.58 units of Indirect Haemagglutinating (IHA) units of antibody against Pasteurella multocida and Log25.44±1.02, Anti Hemolytic Units (AHU) of the antibodies against hemolytic toxins of Clostridium chauvoei. There was no significant difference among the titers of FMDV "O", "A" and "Asia 1"; Pasteurella multocida and Clostridium chauvoei whether used in monovalent or in multivalent.In present study anti-NSP-FMD virus ELISA antibodies in the animals vaccinated with FMD (whole culture) vaccine were undetectable on 15 days post priming while detectable on 30 and 45 days post priming. However anti-NSP-FMD virus ELISA antibodies in the animals vaccinated with FMD (NSP free) vaccine were undetectable on 15, 30 and 45 days post priming. Moreover these antibodies were detectable in FMD carrier animals on 15 days post recovery.Cellular pellet of Pasteurella multocida, Clostridium chauvoei can be used to further minimizing the volume of culture required and further Brucella abortis vaccine can be added in it in conjunction with FMD. This will revolutionize the field of vaccination in Pakistan.
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261.
Immunobiological And Molecular Characterization Of Pasteurella Multocida From Buffaloes
by Muhammad Kamran | Prof. Dr. Mansur-ud-Din Ahmad | Dr. Aftab Ahmad Anjum | Prof. Dr. Azhar.
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Publisher: 2012Dissertation note: Hemorrhagic septicemia is an acute bacterial disease of buffaloes and cattle caused by Pasteurella multocida. In the present study, 400 samples (200 from carriers and 200 from sick animals) from Sargodha division were collected. Among four districts of the division, 15 samples were positive by API Kit, 13 by conventional biochemical tests and eleven were found positive for P. multocida through serological and molecular characterization. Biochemical profile index obtained with API kits had lesser accuracy than conventional and serological profiles for the identification of P. multocida. Passive mouse protection test and AGPT were used for serological confirmation. Different molecular techniques like SDS-PAGE, PCR and RFLP were used to investigate variation at the molecular level in field and vaccinal strains. There were no significant variation between field isolates and vaccinal strain in sick animals and carriers, or in isolates of different districts. Five major and three minor polypeptide bands were observed by SDS-PAGE. Genetic relatedness among the isolates was assessed by cluster analysis using Fingerprint Analysis of Missing Data (FAMD) of 12 isolates. The12 isolates clustered into 5 groups namely I, II, III, IV and V. Group I and II consisted of only one isolate in each (8.33%) of the total designated BKC-01 (S5) and KBO-01 (S1), respectively. Group III composed of 2 isolates (16.67%) namely KBC-02 (S4) and MNO-01 (S2). Group IV had the highest numbers of isolates (50%) designated as KBC-02 (S3), MNO-01 (S6), BKO-02 (S7), MNC-02 (S8), SGO-02 (S9) and V. Only two isolates were typed in group V (16.67%) named as SGO-01 (S10) and BKO-01 (S11).
The size of amplified gene was 460 bp. HindIII I endonuclease cleaved bacterial genome at four sites as compared to other four enzymes (DNase1, PstlI, EcorI and BamHI) change the writing of these enzymes which cleaved at two sites. The isolates were also subjected to ten routinely used antibiotics for sensitivity testing and found enrofloxacin as drug of choice with 90.91% sensitivity, followed by gentamycine, chloramphenicol, ciprofloxacine and norfloxacine (72.73%), ampicillin and amoxycillin (45.45%), amikacin (36.36%) and lowest to sulfadiazine and erythromycine (18.18%).
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262.
Molecular Identification Of Soil Borne Bacillus Anthracis From Districts Lahore And Sheikhupura
by Tariq Jamil | Prof. Dr. Masood Rabbani | Dr. Muhammad Zubair Shabbir | Prof. Dr.
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Publisher: 2013Dissertation note: Background:
Anthrax is a bacterial zoonotic disease caused by Bacillus anthracis. Accurate assays for etiological identification are necessary to ensure proper veterinary and medical health facilities against such diseases. Real-time PCR is a powerful technique to identify this organism based on the presence of two unique plasmids (pXO1 and pXO2) and is highly preferable technique over conventional detection assays in clinical and environmental samples both.
Methodology:
Real Time-PCR technique was used to identify Bacillus anthracis bacteria in the soils of districts Lahore and Sheikhupura. Soil samples were collected from each village of both districts and processed for genome extraction using commercial soil DNA extraction kit. Following genome extraction, the samples were run further for real-time PCR analysis. Positive controls, primers and probes were provided by the Penn state University. SPSS software and pearson's chi square distribution test were used for statistical analysis.
Findings and Suggestions:
Real-time PCR was found as a powerful tool to detect Bacillus anthracis in environmental samples. The bacterium detected was of non-virulent type and showed associations with soil humidity and land use. Further studies may include study of the bacterium with respect to soil-chemistry and sero-prevalence among positive areas of the two districts. Strain characterization is also recommended. The present results may also help in ecological niche modelling by using spatial mapping techniques. Such studies will help in a better understanding of soil as a reservoir for zoonotic organisms and surveillance of the diseases.
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263.
Comparative Analysis Of Respiratiory Microbiota From Clinically Healthy And Deseased Broiler Breeders
by Husnain Ahmed | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad.
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Publisher: 2014Dissertation note: The pulmonary system of birds is a considered a reservoir of innumerable bacterial pathogens including those which are subject to public health significance. This scenario makes respiratory tract of birds prone to many bacterial infections as well. There are many respiratory outbreaks in poultry that causes huge economic losses and a number of bacterial pathogens either acting as primary or secondary pathogen can be held responsible for these losses. A very small fraction of (<1%) of bacterial species are culturable, limited as well as highly variable and time consuming conventional microbiological procedures have typically excluded the normal flora present in the respiratory tract or have restricted the analysis to potential pulmonary pathogens. Due to unculturable nature of many bacterial species there is a very little room left for discovering or determining novel organisms or pathogens and their association with clinical outcomes through conventional microbiological procedures. With the advancement of technology metagenomic analysis of a given sample has emerged as a major culture independent technique for identification of many pathogens, by reading the hypervariable region of the 16S rRNA gene, culture-independent technique such as 454-pyrosequencing, can provide species specific sequence of any bacteria in a given sample.
A total of 12 T-BAL samples from breeder birds were selected based upon the quality and quantity of the double-stranded gDNA. Using 454 bar-coded pyrosequencing, the hypervariable regions of the 16S rRNA gene corresponding to V1 – V5 (~ 1,000 bp) were sequenced. Of the high-quality reads obtained (296,811) using the MOTHUR platform, the sequences were processed for sequence alignment with the 16S RDP database via BLASTn, and subsequent taxonomic analysis through MEGAN programs using a homology-based method to bin sequence reads. The results of study indicate that birds harbor a diverse microbial community including number of phyla, families, genera and characterized bacterial species. The bacterial communities were relatively conserved at the phylum level; however, at lower taxonomic levels, differences were observed in the phylotypes and abundance between the clinically diseased and healthy birds. As indicated by the rarefaction plot and the Shannon-Wiener and Simpson-Reciprocal diversity indices, the biodiversity and richness in the taxonomic content was higher in the clinically healthy birds compared with the diseased birds. Regardless of the bird health status a number of new species were identified. A number of these bacterial species have been found to be associated with infectious diseases in humans and other species, although the clinical importance of these bacteria as part of the respiratory microbiome of birds has not been established.
As the nature of bacterial species is to constantly act with one another and, potentially, with the birds themselves provides an interesting avenue for continued research. There is a need to conduct further clinico-pathological studies to establish the link between causes versus effects.
Availability: Items available for loan: UVAS Library [Call number: 1826,T] (1).
264.
Evaluation Of Food Safety Of Common Salads And Antimicrobial Activity Of Natural Dressings
by Awais Fida | Dr. Arfan Ahmad | Dr. Ali Ahmad.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Salad is a commonly consumed raw food which is equally favored by each group of the society. The consumption of these salads among common man has been increased in recent past. Several outbreaks have been reported throughout the world regarding to GIT tract infections and isolation of thermotolerent FC. Coliforms or especially Fecal coliforms are considered indicator organism whose absence ensures the lack of non pathogenic bacteria.
The objectives of the present study was to evaluate the sanitary condition followed by food practitioners in Lahore area and the effect of natural salad dressings with respect to varing concentrations of NSDs, at different temperature and interval of exposure.
A total of 60 samples were purchased from local market, 12 of the samples were collected from each selected area (i.e. Anarkali, Gawalmandi, Samanabad, Sanat Nagar and Shad Bagh) while the sample purchased from each area is from 4 different shops (3 from each) i.e. Burgar salad, Gravey Chana salad, Halwa Puri salad and Bar-B-Q salad shops.
The samples were transported to UDL, UVAS, Lahore under cooled chain and subjected for analysis to enumerate the quality by estimating FC using MPN method (FDA 2010). Samples are divided into 4 sections. One was taken as control and the others were taken as experimental and subjected to various concentrations of NSD's, after exposing to 5, 15 and 30min to antimicrobial agents the samples are transmitted to triplet of test tubes containing EC broth. The inoculated tubes were provided with 24-48 hours incubation at 44.5oC.
The gas production in tubes indicted the presence of FC, so values were calculated using MPN table (Annexure XXV) and log values were written to get log reduction values.
More than 70% salad samples collected from selected areas were found unhealthy for consumption. Maximum contamination was observed in salads which are being served with night meals (i.e BBQ salad) while Anarkali was found an area with maximum of positive samples. The contaminations were least recorded in winters while maximum in rainy season. Among the NSD's Vinegar was considered best antimicrobial agent followed by Lemon and Garlic but statistically no significant difference between these two were observed. There was no significant relationship observed between temperature-time, concentration-time and temperature-concentration of NSD.
Availability: Items available for loan: UVAS Library [Call number: 1829,T] (1).
265.
Identification Of Multiple Drug Resistant (Mrd) Mastitis Causing Bacteria In Dairy Goats
by Muhammad Faisal najees | Dr. Aftab ahmad anjum | Prof. Dr. mansur-ud-Din ahmad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1852,T] (1).
266.
Isolation Characterization And Growth Optimization Of Starch Hydrolyzing Fungi From Soil Of Livestock Farms
by Saba Sana | Prof. Dr. Aftab ahmad anjum | Dr. Muhammad Nawaz | Prof. Dr.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1898,T] (1).
267.
Study On Micobiological Quality Of Water Supplied To Poultry Birds From Tube Wells Water Pumps Drilled Up To Varying Bore
by Sidra Moqddes | Prof. Dr. Masood rabbani | Dr. Jawad Nazir | DR.Yasin tipu.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1922,T] (1).
268.
Plasmid Mediated Analyses And Plasmid Curing Of Previously Isolatedmulti-Drug Resistant Eschetichia Coli From Retail
by Mawra gohar | Dr. Ali ahmad sheikh | Dr.Tanveer | Prof, Dr. Aftab ahmad anjum.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1923,T] (1).
269.
Physico-Chemical Factors Affecting The Growth Of Bovine Rotavirus
by Wardah sharmeen syed | Prof. Dr. Tahir yayub | Dr Muhammad Zubair shabbir | Dr.Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1924,T] (1).
270.
Seroprevalence Of Bluetongue In Domestic Animals
by Farid ahmed khan | Prof..Dr. Khushi Muhammad | Ms. Sehrish | Prof. Dr Tahir yaqub.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1925,T] (1).
271.
Isolation And Molecular Identification Of H9N2 Avian Influena Virus From Human In Punjab Province Pakistan
by Abdul ahad | Prof .Dr, Masood rabbani | Prof. Dr. Rana | Prof. Dr. Tahir yaqub.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1926,T] (1).
272.
Role Of Water Chemistryand Stabilizers On The Infectivity Of Virus In Live Neqcastle Disease Virus Vaccine
by Sahrish tariq | Dr Jawad Nazir | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1927,T] (1).
273.
Bacillus Thuringiensis Toxins Biological Control Aginst Aedes Aegypti
by Qurat ul ain hanif | Prof. Dr Tahir yayub | Dr.Sehrish firyal | Prof. Dr.Khushi Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1929,T] (1).
274.
Characterization And Use Of Phytase Producing Bacterial Isolates To Enhance The Nuteitive Value Of Poultry Feed
by Sohail Aslam | Prof, Dr. Ahmad anjum | Dr. Imran Altaf | Dr. Muhammad Wasim.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1930,T] (1).
275.
Evaluation Of Probiotic Potential Of Locally Characterized Lactobacillus Spp. In Broiler
by Saima asghar | Dr.Muhammad Nawaz | Mr. Muhammad Asad ali | Prof. Dr.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1931,T] (1).
276.
Newcastle Disease Virus Act As An Adjuvant For Antibody Response Of Broilers To Inactivated Mycoplasama Gallisepticum Vaccine
by Rabia Riaz | Prof. Dr. Khushi Muhammad | Dr. Muhammad | Dr.Ali ahmad sheikh.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1932,T] (1).
277.
Antibody Response Of Goats To Gel Based Combined Vaccine Against Peste Des Petits Ruminants Contagious Caprine
by Muhammad Khalil | Prof.Dr. Khushi Muhammad | Dr. Aneela Zameer Durrani | Dr.Jawad Nazir.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1933,T] (1).
278.
Microbiological Quality Assessment And Antibiotic Resistance Of Microbes Present In Fresh Indigenous Juices And Milk
by Hafza Raahat Farooq | Prof. Dr.Aftab ahmad Anjum | Prof. Dr. Khushi Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1934,T] (1).
279.
Correlation Of Deifferent Managment Systems And Facilities Of Retail Milk Shops With That Of Microbial Load In Raw And Pasteurized Milk
by Faria kanwal | Prof. Dr. Masood rabbani | Dr. Ali ahmad sheikh | Dr.Wasim.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1953,T] (1).
280.
Bacterial Growth Inhibition Based Detection Of Beta-Lactam Antibiotic Residues In Milk
by Sana moin | Prif. Dr. Masood rabbani | Dr Ali ahmad | Dr. Yasin tipu.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1954,T] (1).
281.
Characterization Of Beta-Lactamase Producing Genes From The Chicken Meatborne Antimicrobial Resistant
by Ayesha tabassum | Dr. Ali ahmad sheikh | Prof. Dr.Masood rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1955,T] (1).
282.
Detection Of Gram Positive Foodborne Pathogen From Retail Quail Meat Through Optimized Multiplex Pcr
by Iqra Safdar | DR. Ali ahmad sheikh | Dr.Tanveeer | Ms.Fareeha akhtar.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1956,T] (1).
283.
Detection Of Gram Negative Foodborne Pathogen From Retail Quail Meat Through Potimized Multplex Pcr
by Amna kanwal | Dr.Aki ahmad sheikh | Dr.Tanveer | Prof, Dr. Masood rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1957,T] (1).
284.
Microbiological And Physiochemical Analysis Of Drinking Water From Human And Veterinary Hospitals
by Kiran batool | DR. Arfan ahmad | Dr Hassan mushtaq | DR. jawad nazir.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1958,T] (1).
285.
Evaluation Of Drinking Water Quality At Zoo And Various Public Places In Disteict Lahore
by Maimona jibreel | Dr. Arfan ahmad | Dr.Hassan | Prof. Dr. Masood rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1959,T] (1).
286.
Antibody Resoinse Of Broilers To Oil Based Combined Mycoplasma Gallisepticum And Avian Influenza H9
by Sadi Sarfaraz | Prof. Dr. Khushi Muhammad | Prof.Dr. Asim | Prof.Dr.Tahir yaqub.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1976,T] (1).
287.
Isolation Characterization And Optimization Of Potential Probiotic Bacteria From Poultry Droopings
by Muhammad Hashim khan | Prof. Dr. Aftab ahmad anjum | Dr. Jawad nazir | Prof. Dr. Mansur-ud-din.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1991,T] (1).
288.
Comparative Study On Prevalence And Antimicrobial Susceptibility Pattern Of Extendedspectrum B- Lactamase
by Karam rasool | Prof. Dr. Masood rabbani | Dr. Ali ahmad sheikh | Dr. Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1992,T] (1).
289.
Screening And Characterization Of Phytase And Bile Salt Hydrolases Producing Probiotic Lactobacilli Isolated
by Madiha arif | Dr. Muhammad Nawaz | Prof. Dr. Khushi muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2001,T] (1).
290.
Isolation And Characterization Of Antibiotic Resistant Lactobacilli From Fermented Food Products
by Shahgull | Dr. Muhammad Nawaz | Prof. Dr | Prof. Dr. Aftab anjum.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2002,T] (1).
291.
Screening Of Indigenous Microalgae For Antimicrobial Activities And Growth Optimicrobial For Mass Production
by Imran hanif | Prof. Aftab ahmad anjum | Dr. Jawad nazir | Ms. Sehrish firyal.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2015,T] (1).
292.
Production Of Hyperimmune Yolk Against K99 Enterotoxigenic E. Coli Purification Of Lgy
by Khadija abid | Prof. Dr. Tahir yayub | dr. M. wasim | Prof. Dr.Khushi muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2057,T] (1).
293.
Effect Of Temperature And Relative Humidity On The Survival Of Newcastle Disease Virus Isolates Using Germ Carrier Techniques
by Tayyeba Sohail (2009-VA-209) | Dr. Jawad Nazir | Prof. Dr. Khushi Muhammad) | Prof. Dr. Asim Aslam.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Newcastle Disease (ND) is a highly contagious viral disease that affects almost all avian species including poultry, cage and wild birds around the globe (Terregino et al. 2003; Vidanovic et al. 2011). ND is economically important disease and included as list-A disease of Office des International Epizootics (OIE) (Anonymous). Mortality of infected birds ranges from negligible to as high as 100 % depending on the pathotype of the virus involved and health status of the birds (Alexander and Manvell 2004). NDV is an enveloped virus with single stranded, non-segmented, negative sense RNA genome (Makoui et al. 2013). The virus belongs to Avulavirus genus of Paramyxoviridae. There exist only one serotype of NDV designated as avian paramyxovirus-1 (Kapczynski et al. 2013) however, different virus strains do vary in their pathogenicity. There are 3 pathotypes of NDV; velogenic (highly virulent), mesogenic (moderate virulent), and lentogenic (mild virulent) based upon diseases producing potential and severity of signs in the infected birds (de Leeuw and Peeters 1999).
NDV is primarily transmitted to the susceptible birds through aerosol and fecal oral route (Martin 1992). Infected birds secrete high amount of the virus in their feces, saliva, mucous and nasal secretions which might contaminate the premises. Inanimate objects or fomites are a potential reservoir of viruses outside the host and might play an important role in the transmission of pathogens (Nicas and Sun 2006). Several factors can influence the survival of viruses outside the host (Sobsey and Meschke 2003; Weber and Stilianakis 2008; Stallknecht and Brown 2009). A number of studies show that respiratory pathogens can survive from hours to months on fomites (Abad et al. 2001; Kramer et al. 2006). Certain physical factors like temperature, humidity, pH, salinity, exposure to ultraviolet (UV) rays etc drastically affects the
Introduction
2
virus persistence in the environment. Effect of such physical insults is more pronounced on enveloped viruses than non-enveloped ones (Mbithi et al. 1991; Schaap et al. 2012; Tuladhar et al. 2012). High humidity and temperatures not only reduces the survival of influenza viruses on contaminated surfaces but also modulates their transmission to the susceptible birds (Shaman and Kohn 2009; McDevitt et al. 2010; Paynter 2014). Similarly lower temperature and less humidity promote the survival of NDV in the environment (Dat and Chuc 1985; Kournikakis et al. 1988).
ND is endemic in Pakistan but since last few years several new virus strains are circulating in commercial and rural poultry of the country (Munir et al. 2012; Shabbir et al. 2013). Central Punjab region is densely populated with commercial poultry and serve as disease epicenter every year. It has been observed that the disease outbreaks usually start in December, attain peak in the late winter and spring season, start decline in June and disappear in the rainy season. Apart from several other contributing factors, environmental survival of the viruses might contribute to the disease outbreaks. Availability: Items available for loan: UVAS Library [Call number: 2227-T] (1).
294.
Prevelance Of Brucellosis In Aborted Women Visiting Tertiary Care Hospitals Of Lahore City
by Saba Yasmin (2009-VA-211) | Prof. Dr. Aftab Ahmad Anjum | Dr. Tayyaba Ijaz (Co Supervisor) | Prof. Dr. Khushi Muhammad | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pakistan is an agriculture based country whose rural population depends upon livestock for livelihood. Contribution of livestock to agriculture sector is 55.9 percent while 11.8 percent to the national GDP during 2013-14 (GOP 2013-2014). A number of infectious diseases hamper the growth of livestock sector. Some of the livestock diseases are zoonotic in nature and threat to human health. Brucellosis is considered among major zoonotic diseases throughout the world. The Mediterranean Basin, south and Central America, Eastern Europe, Asia, Africa, the Caribbean and the Middle East are considered as high-risk countries (Memish 2001).
Brucellosis in human beings is a major concern of community health. It causes acute and chronic illness, physical incapacity and loss of health. Bacterial species involved include Brucella abortus, Brucella melitensis or Brucella suis. Brucellosis is acquired by human beings from infected animals by close contact with vaginal secretions, urine, feces, blood, aborted fetus, or consumption of unpasteurized milk or other raw milk products. Shepherds, milkmen, butchers, knackers, veterinary assistants and abattoir workers are at high risk (Agasthya et al. 2007). Prevalence of brucellosis recorded by Mukhtar and Kokab (2008) in abattoir workers of Lahore Pakistan was 21.7 percent. Higher prevalence of brucellosis was observed in females (37.06%) than males (24.2%) in patients admitted at Peshawar, Pakistan (Shahid et al. 2014).
Symptoms of disease vary among human patients, ranging from non–specific, flu-like symptoms (acute form) to undulant fever (chronic form). Some of the serious complications of skeletal system, cardiovascular and central nervous systems may develop. Other important signs observed include arthritis, orchitis, epididymitis, abortion, retained placenta and stillbirth (Baba et al. 2001; Grilló et al. 2006). In animals, brucellosis in most of the cases results in abortion, birth of weak calves, death of young stock, infertility in males and reduced milk yield in females (Maadi et al. 2011; Abubakar et al. 2012).
There is actual need for teamwork between public health officials and veterinary officers to reduce communication of brucellosis between animals and human in endemic areas (Jelastopulu et al. 2008; Makis et al. 2008). Clinical picture of brucellosis is nonspecific and may vary from patient to patient. Therefore, laboratory diagnosis by isolation and culture or recognition of specific anti–Brucella antibodies is essential for confirmation of brucellosis (Al-Attas et al. 2000).
Diagnosis of brucellosis by culture and phenotypic description is time-consuming. Furthermore, risk of infection to worker is always there. Serological tests are commonly preferred for brucellosis in cattle and small ruminants, especially at farm level screening. Chance of cross-reactions with other gram negative bacteria is a major problem. Rose Bengal Plate Agglutination Test (RBPT) and Slow Agglutination Test (SAT) are extensively used for detection of anti-Brucella antibodies (Halling et al. 2005). Enzyme Linked Immunosorbent Assays (ELISA) have been developed to resolve suspected samples by RBPT. ELISA is more sensitive, so it can detect Brucella carriers which are negative by RBT, SAT and CFT (Aert et al. 1984). Molecular techniques are more reliable and specific than serological tests. Final confirmation of brucellosis is carried out using polymerase chain reaction (PCR), a molecular technique. Real-time PCR offers enhanced sensitivity, specificity and rapidity of performance when compared to conventional PCR (Gwida et al. 2012). Availability: Items available for loan: UVAS Library [Call number: 2225-T] (1).
295.
Modulation Of Antibiotics Resistance Pattern In Escherichia Coli By Different Plant
by Bushra Chaudary (2009-VA-232) | Dr.Muhammad Nawaz | Prof. Dr. Aftab Ahmed | Dr. Naureen Naeem.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Escherichia coli (E. coli) is Gram negative microorganism belonging to family
Enterobacteriaceae. It is part of normal micro flora of gastrointestinal tract of human and all
warm blooded animals (Kaper et al. 2004). Escherichia coli is source of many infectious diseases
in human as well as in animals. Common E. coli infections are enteritis, urinary tract infection,
septicemia and neonatal meningitis. In pets and farm animals, E. coli is associated with diarrhea
(Allocati et al. 2013). Poultry industry is facing huge annual losses due to infection of avian
Pathogenic E. coli (APEC) in broilers (Oosterik et al. 2014). E. coli causes a variety of
syndromes in poultry including yolk sac infection, respiratory tract infection, swollen head
syndrome, septicemia and cellulitis (Buys et al. 1989)
Antibiotics are chemical agents which inhibit the microbial growth and used to eradicate
infections. Mechanisms of action of antibiotics provide a base to categorize antimicrobial agents.
Most important classes of antibiotics act as inhibitors of cell wall synthesis, protein synthesis
(tetracyclines and macrolides), nucleic acid synthesis (fluoroquinolones), metabolic pathway
(trimethoprim-sulfamethoxazole) and cell membrane (polymyxins). Bacteria may have intrinsic
or acquired resistance to antimicrobials (Tenover 2006).
Urinary tract infections are mostly caused by E.coli. Antibiotics generally used for the treatment
of E. coli infections include ampicillin, nitrofurntion, cephalosporin, sulphonamides
(trimethoprim-sulfamethoxazole) and quonolones (neladixic acid, ofloxacine, ciprofloxacin and
levofloxacine) (Lin and Lin 2010).
Extended use and misuse of antibiotics lead to the development of resistant bacteria. Resistant
E. coli strains are common source of hospital born and community acquired infections. Ease of
Introduction
2
international travelling is one of the major spreading factor for antibiotic resistance. Resistant
bacteria got opportunity to move from one geographical area to another (van der Bij and Pitout
2012). New strains of E. coli resistant to carbapenems (New Delhi metallo-β-lactamase 1 (NDM-
1) are major global health issue (Kumarasamy et al. 2010). Antibiotic resistance has become a
serious public health problem. Currently, world is facing great difficulty in treatment of many
infectious disease of human and animals. One of the reasons of treatment failure is emergence of
resistant bacteria (Levy 2002).
To develop new strategies for treatment of infectious diseases, it is necessary to understand the
mechanisms of resistance. Efflux pump inhibitors, enzymatic degradations and alteration of
target sites are major strategies by which bacteria acquire or develop resistance to antibiotics
(Sibanda and Okoh 2007).
Scientists are looking for alternatives of antibiotics such as bacteriopheges, naturally
antimicrobial compounds and some non antimicrobial agents (Worthington and Melander 2013).
Probiotics (Lactobacillus and bifidobacterium) can be a prophylactic measures against E. coli
and may be used to treat intestinal tract infections of E. coli and other bacteria (de Vrese and
Schrezenmeir 2008).
Phytochemicals, secondary metabolites of plants, have antibacterial activity against many
pathogenic organisms. These phytochemicals in combination with antibiotics may show
synergistic effect. Phytochemicals and plant extracts can be a source of antibiotic resistancemodifying
agents (RMAs) (Abreu et al. 2012). Plant extracts shown antibacterial activity
because of phytochemicals like alkaloids, tannins, flavonoids, phenolic compounds and steroids
(Gobalakrishnan et al. 2013). Plant extracts are used as traditional medicine for the treatment of
many diseases. Plant extracts like Zingiber officinalis (Ginger) Gymnema sylvestre (Gurmar buti), Astragalus (goat’s thorn), Calotropis procera (apple of Sodom) and oputia dillenii (cactus)
have antimicrobial activity (indu et al. 2006 and Kumaar et al. 2013). Plant extracts also have
antibiotic resistance modulation potential (Mako et al. 2012).
Availability: Items available for loan: UVAS Library [Call number: 2247-T] (1).
296.
Efficacy of Various Disinfectants Against Newcastle Disease Virus Isolates In Relation To Different Temperatures
by Momena Habib | Dr. Jawad Nazir | Prof.Dr.Aftab Ahmed Anjum | Dr. Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Newcastle disease (ND) is an infectious viral disease of birds and has been considered as one of the most significant diseases of poultry and other avian species around the globe (Aldous and Alexander 2001). The disease is endemic in Pakistan and causes major economic losses to poultry industry each year (Rehman H et al. 2013). Although incidence of ND in broiler and layers remains higher during the course of year, it reaches its plateau during seasonal stress (January-February and June-July). One report shows that during September 2011 - January 2012, the diseases has killed 45 million birds and resulted into a loss of approximately 65 million US $ to the country (Anonymous). ND virus (NDV) is an enveloped, single stranded RNA virus and grouped into the genus Avulavirus within family Paramyxoviridae(Calibeo-Hayes et al. 2003). Isolates of NDV may be categorized into three main pathotypes (lentogenic, mesogenic and velogenic) depending upon the severity of disease in chickens (Seal et al. 2000).
NDV is most commonly transmitted through fecal-oral and respiratory route provided that birds are in close contact (Alexander 1988). Infected birds excrete large amount of virus in their feces (Calibeo-Hayes et al. 2003). Virus is present in most tissue secretions of acutely infected birds even 24 hours prior to appearance of clinical signs. Under certain conditions, NDV can survive up to 20 days in fecal material (anonymous) and presence of organic matter might enhance the virus survival rates (Clarke et al. 1956). In order to control the spread of disease, decontaminate the infected premises is of prime importance. Strict biosecurity measures in conjunction with mass vaccination programs are also employed for the control of the disease. In past few decades, implementation of extensive vaccination programs in commercial poultry has reduced the
Introduction
2
disease incidence to some extent in Pakistan. However, this might also led to the generation of novel genotypes under high immune pressure(Miller et al. 2009). Disinfection of farm premises plays significant role to break the disease cycle of microbial pathogens(Fawzia1 et al. 2013). Such procedures gain more significance on the farms that experience disease outbreaks. Dry cleaning, burning and fumigation followed by chemical disinfection is routinely practiced in Pakistan. Although, physical method are helpful to reduce the pathogen load on farm premises but complete disinfection cannot be achieved without the use of chemical disinfectants. Commercial disinfectants employed in Pakistan are grouped into oxidizing agents, aldehydes, quaternary ammonium compounds, and phenolic preparations. The products containing these ingredients are available either alone or various combinations under different brand names. (Stringfellow et al. 2009).
All of the disinfectants do vary in their disinfection power but to achieve proper disinfectants appropriate interaction time is necessary. Under clean conditions such interaction time is low but longer contact times are needed in presence of organic matter. Some studies show that environmental temperature influences the interaction time of disinfects with matter/surfaces to achieve proper disinfection. In case of chlorine disinfection of adenovirus inactivation time was increased 2-3 times when temperature was lowered to an extent of 10°C (Clarke et al. 1956). Similarly, virucidal activity of some disinfectants including aldehyde, chlorine and iodine compounds against infectious bursaI disease virus (IBDV) was also decreased at lower temperatures (Benton et al. 1967). Availability: Items available for loan: UVAS Library [Call number: 2226-T] (1).
297.
Occurrence Of Bacterial Contaminants In Poultry Meals And Their Antibiotic Resistance Pattern
by Nayyab Tariq (2009-VA-207) | Prof. Dr. Aftab Ahmad Anjum | Dr. Muhammad Nawaz | Dr. Muhammad Nasir.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Poultry is the second largest industry after textile industry in Pakistan. Its consumption
rate is very high as compared to other animal protein sources, as it is cheaper as compared to red
meat. To fulfill increasing demand of poultry, poultry production quality must be improved.
Many factors affect poultry production. One factor is feeding process. Efficiency of poultry
production depends mainly on feeding process which influences both the quality and quantity of
the poultry production (Grepay 2009). The rearing of poultry birds on commercial level requires
use of bulk quantities of poultry feed. Poultry feed costs 60-70% of total cost for production
(Sahraei et al. 2012). The main purpose to increase poultry production is to fulfill nutritional
requirements of human population that largely rely on poultry and poultry by products as a
source of protein(Obi and Ozugbo 2007).
Poultry feeds are food materials designed to contain all necessary feed ingredients for
proper growth, meat and egg production in birds (Obi and Ozugbo 2007). It is a mixture of
various components including plant proteins (cereals and by products, grains etc), animal byproducts,
fats, vitamins and minerals (Ravindran 2013). The major component of poultry feed is
protein which is the key component of eggs and meat. Protein sources in poultry feed are of
plant, marine and animal origin. Plant proteins may lack some of the essential amino acids, thus
are incomplete protein. Proteins of animal origin are better growth promoter than protein of plant
origin, but their safety is a concern. Among plant based proteins, soybean and canola meal are
produced in higher amounts worldwide (Alali et al. 2011). The animal protein sources include
poultry, fish, meat bone and poultry by products meal. Poultry meal is derived from clean tissues
Introduction
2
of slaughtered poultry including bone after the moisture and fat have been extracted in the
rendering process. It may contain whole birds excluding feathers (Anonymus 2014). Among all
protein based meals, poultry meals and poultry by products meal are of superior quality and
provide higher protein content than plant, marine and meat based meals (Samli et al. 2006).
Quality of animal feed has gained importance worldwide. The feeds are found to be
associated with infectious or non-infectious hazards, thus influence human health (Sherazi et al.
2015). Poultry feed can act as carrier of animal and human pathogens (Aliyu et al. 2012). Poultry
feed can get contaminated at any point of harvesting, processing, storage or dispersal of feed.
Primary mode of poultry feed contamination is by dust, soil, water and insects. Poultry meals can
be another source of feed contamination. Poultry meals are added in feed as a source of protein.
Feeds of animal origin like poultry meals are richer in nutrients and water as compared to feed of
plant origin thus are found to have higher microbial load, facilitating the multiplication of
bacteria (Kukier and Kwiatek 2011). Inclusion of contaminated meals in feed increases microbial
load of poultry feed. The contamination of poultry feed not only influences appearance and
nutritional value of feed, but also affects animals and human who consumes it (Maciorowski et
al. 2007). The profitability of poultry production can be greatly affected due to the frequency of
feed contamination and the detrimental effects of the aflatoxins on performance of chickens
(Anjum et al. 2011). Poultry feeds have been implicated in several poultry diseases of viral
(Avian Influenza, Newcastle disease), bacterial (Salmonellosis, Infectious Coryza) and fungal
origin. Many human diseases like Traveler’s Diarrhea and Salmonella Paratyphoid fever have
been associated with consumption of poultry birds that contracted infections from poultry feed
(Obi and Ozugbo 2007).
Introduction
3
The poultry industry relies on ready to use poultry feed prepared by feed mills (Arotupin
et al. 2007). Both bacteria and fungi including mycotoxins usually contaminate feed at different
stages of pre or post processing, depending upon the conditions under which it is handled or
stored (D’Mello 2006). Poultry meals mostly get contaminated post rendering process. The
cooking step in rendering process inactivates bacteria, viruses, protozoa, and parasites(Meeker
and Hamilton 2006) . Still presence of contaminants in meals is attributed to post processing
contamination. Many bacterial pathogens reported in feed are Escherichia coli, Erwinia
herbicola, Salmonella spp., Listeria spp., Enterococcus fecalis, Cl. perferingens and Cl.
botulinum (Aliyu et al. 2012; Lateef and Gueguim-Kana 2014) . The contaminated feed results
in excessive activation of immune system and ultimately decreases poultry production and its
profitability (Kukier et al. 2012). In addition to bacterial contaminants, toxigenic fungi have
threatened quality and safety of feed and have caused severe losses to poultry industry in recent
times. Cereals and grains based poultry feed mostly get contaminated with fungi (Kwiatek and
Kukier 2008). Mycotoxin producing fungal genera that are reported in poultry feed are
Aspergillus, Penicillium and Fusarium (Greco et al. 2014).
As Poultry feed is the first step of the food safety chain in "farm-to-fork" model. Contaminated
feed can also serve as a source of antimicrobial resistant bacteria in poultry meat(da Costa et al.
2007). There are many evidences that pathogens in feed are transmitted to humans through
animals and food of animal origin. It can also become source of some human pathogens in
environment. Feed contamination by fungi is responsible for animal mycotoxicoses and through
consumption of contaminated animal food, results in human intoxications (Kukier et al. 2012).
Birds utilizing toxins containing feed are economical loss for farmers and also affects consumer
Introduction
4
health through its residues (Alam et al. 2012). Poultry feeds containing antibiotic resistant
bacteria results in loss of poultry productivity, making treatment of poultry diseases difficult.
Thus quality of animal food directly depends on usage of nutritionally balanced and safe feed.
Among many feed sources used, poultry meals are gaining importance for their higher nutritional
value, but very less work has been done in world particularly in Pakistan to determine
microbiological safety of poultry meals produced. There is the need to determine various quality
parameters which should be followed to ensure production of safe meal. Availability: Items available for loan: UVAS Library [Call number: 2252-T] (1).
298.
Microbiological Quality Assessment Of Human And Veterinary Drugs
by Sadaf Riaz (2007-va-277) | Dr. Ali Ahmad Sheikh | Dr. Fareeha Akhtar | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The continued increase in population results in increasing demand for pharmaceutical products. According to Pakistan pharmaceutical manufacturers association, Pakistan has approx. 400 pharmaceutical industries including 25 multinational industries. The Pakistan pharmaceutical industry meets 70% of country’s demand of pharmaceutical drugs.
Medicines are chemical compound that are administrated to human or animal as an aid to diagnosis, treatment or prevention of disease (Lecca, 1978).
Microbiological quality assessment is very important point of pharmaceutical manufacturing. The term quality in its wide sense means “Safety “. Microbial contamination means, presence of undesired micro-organisms or their metabolic products (Uba, 1990).
Pharmaceutical drugs are used in many ways in treatment of diseases (Mugoyela et al. 2010). Pharmaceuticals of different forms are susceptible to contamination by different microbes (Aulton, 2002). Contamination of medicines with micro-organisms can bring about changes in their physiochemical properties, results in product degradation before their expiry date (Shaikh et al. 2000). Microbial contamination rang from true pathogen like Clostridium tetani, to opportunistic pathogens, such as Pseudomonas aeruginosa (Mwambete, 2009). Microbial infections not only due to physical presence of micro-organisms, but also their metabolites/toxins can be very harmful even in low quantities (Shukla et al. 2004). Medicines used in treatments cannot afford to have microbial contamination. Patients who take medicines have very weak immune system which makes them vulnerable to wide range of infections. Microbial contamination of drugs may contribute to secondary microbial infections in immunologically weak patients (Adeshina et al. 2009). Contaminated pharmaceuticals may lead to medication complicacy in patients.
Drugs can be divided into two types on the basis of microbiology; sterile pharmaceutical products and non-sterile pharmaceutical products (Clement et al. 2013). Sterile pharmaceutical products should not contain any viable micro-organisms and non-sterile pharmaceutical products may contain viable micro-organisms but it should be within official limit and all drugs should not have any pathogenic bacteria. Most of drugs can be contaminated during storage and handling (Takon and Antai, 2006). Antimicrobial agents are added to medicines to minimize microbial growth but should not be added to mask poor manufacturing process (Gad et al. 2011).
The level of microbial contamination in medicines depends on availability of nutrients, water, presence of other micro-organisms, osmotic pressure and oxygen etc. The factors determining the results of drug-borne infections may include the type and amount of microbes, patient’s immune system and route of administration (Baird, 2004).
In recent years manufacturing of drugs have improved. The occurrence of microbes in drugs has been well documented. There have been many reports of infections caused by contaminated drugs (Ibezim, 2002; Coker, 2005). But majority of cases of infections caused by medicines are not recognized and reported as such (Denyer and Baird, 2006). Previous studies have showed microbiological quality concern related to commercially available drugs (Denyer and Baird, 2006).
Contamination of drugs results in loss of faith of a consumer in pharmaceutical industry and sales would go down (Mugoyela and Mwambete, 2010).
Although pharmaceutical industries are one of the growing and expanding sector in Pakistan, but the quality of medicines varies as they are retail oriented. Unfortunate situation is present in sales of drugs. A number of unlicensed drugs stores selling poorly manufactured drugs
2
Hence the microbiological quality of pharmaceuticals, assessment of number and type of bacteria and fungi within drugs, is essential to ensure consumer safety (Urmi et al. 2014; Tamalli et al. 2013)
This study to relate to national need and reflects its importance. New international trend focus on the quality of pharmaceutical products, unfortunately few studies has been carried out in this prospective which is not proven to be sufficient in predicting the quality of drugs. This has compelled me to do research on this topic in order to assess the level of such microbial contaminants in pharmaceuticals given to patients in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2275-T] (1).
299.
In Process Quality Control Factors Affecting The Quality Of Locally Prepared Salmonella Gallinarum Antigen
by Zahra Malik (2009-VA-245) | Dr. Arfan Ahmad | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Asim Aslam.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Fowl typhoid is a septicaemic disease caused by S. gallinarum biovar gallinarum has major economic significance in many parts of the world. It is an acute or chronic septicaemic disease that usually affects the birds (mostly adult birds). Eradication of disease is normally done by identifying the infected flocks and eliminating the reactor birds by using serological tests, but diagnosis of the disease is much expensive because antigen used for this purpose is imported. The study, therefore, has been proposed to prepare and evaluate the stained antigen of S. gallinarum using local isolates.
A total of 15 isolates were procured from Poultry Research Institute (PRI) Rawalpindi, University Diagnostic Lab (UDL) and Department of Microbiology, UVAS Lahore, which were identified by Biochemical testing and further confirmed by Polymerase Chain Reaction. Among all 15 isolates two isolates were confirmed as S. gallinarum and proceeded to prepare local antigen of S. gallinarum. Locally prepared antigen was checked with known positive and negative sera, Effect of different preservatives (Sodium azide and Thiomersal sodium) and different storage temperatures (4°C, 25°C and -20°C) was also studied after every fifteen days post storage upto 6 months to observe the stability and shelf life of local antigen. On the end of study both preservatives i.e. Sodium azide and Thiomersal sodium was found equally effective for antigen activity, whereas 4°C proved best storage temperature to be used for the antigen preservation.
Activity of locally prepared antigens was also compared with the imported antigen (Charles, River, USA) stored at different temperatures regularly throughout the six months, which showed that local antigens was almost as good as the imported antigen.
Summary
51
CONCLUSION
Locally prepared S. gallinarum antigen was found as effective as imported antigen. Both the test preservatives (Sodium azide and Thiomersal Sodium) had the same effect on antigen preservation. Among all three test temperatures, 4°C was accepted as best storage temperature for the long term preservation of local antigen with either of the preservative. Availability: Items available for loan: UVAS Library [Call number: 2278-T] (1).
300.
Impact Analysis Of Quality Control Practices In Selected Microbiology Based Veterinary Diagnostic Labs Operational In District Lahore
by Faiza Marrium (2009-VA-239) | Prof. Dr. Masood Rabbani | Dr. Ali Ahmed Sheikh | Dr. Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Veterinary diagnostic labs are the powerful ally for the diagnosis, prevention and monitoring of animal diseases in any country. Labs are ordered for test for many reasons. Errors could be present in these tests. So to avoid detrimental effects of these errors a quality control system is required. This system is maintained by following the standards given by recognized international organizations like ISO, OIE, WHO, FAO and CDC etc. These authorities make the standards which should be followed by the VDLs to improve their quality of tests and management to give precise and accurate results which will help them in being well reputed lab which give internationally accepted quality of results.
In this study 4 private and 4 public sectors Veterinary diagnostic Labs was selected and it was assured to labs that their information will remain confidential as data was used only for the research purposes. Labs were identified by the codes given to them for study purpose. Information required for this study was gained through a questionnaire. Information regarding identity of lab, contact numbers, location and type of testing/diagnosis was gathered. Information about the parameters of quality control like building design, power backup, sections of lab, operating equipments, development of log books, availability of certificate of analysis for chemicals, availability of material safety data sheet was gathered. Quality assurance issues were also addressed by gathering information about internal quality assurance program, proficiency testing etc. Then this data was analyzed by using SPSS to interpret the results.
The data was analyzed statistically through frequency distribution by using Statistical Package for Social Sciences (SPSS) version 18.0 for development of Graphs and Tables.
Summary
53
requirements about personnel and equipments were 80% and 87.5% while minimum values were 40% and 25% respectively. Maximum value about quality control measures and waste management were 89.47% and 70% and minimum percentages were 36.40% and 40% respectively. Results have shown that 100% requirements of environmental monitoring and customer care were fulfilled by some labs while some labs only fulfill 20% of these parameters.
Conclusion
This study shows that in Lahore district veterinary diagnostic labs are not giving proper attention to quality of their system and there is no significant difference between setups of private and public sector laboratories. Availability: Items available for loan: UVAS Library [Call number: 2277-T] (1).
