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1. Characterization Of Indigenious Species Of Mycotoxins Producing Aspergilli

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

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

2. Monitoring Of Mixed Vegetable Salads For Microbial Quality

by Sana Hameed | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 2012Dissertation note: The present research was planned to investigate the microbial load and identification of bacteria in mixed vegetable salads from three different locations such as road side vendors, fast food outlet and family restaurants. Total 90 samples were collected 30 from each location. Samples collected carefully in sterilized plastic bags and processed in microbiological lab of UV AS Lahore. The results were compared with different food standards given by developed countries like UK. In present study results showed that for aerobic plate count 30 samples from road side vendors was in range of 105_ 1014, fast food outlets range of 105_ 1012 and family restaurants range of 105_ 1013. Coliform count from road side venders was in range of 105_ 1013, fast food outlets range of 105_1012 and family restaurants range of 105 - 1013. Fungal count from road side vendors was in range of 103_106, in fast food out lets range of 103 - 106 and in family restaurants range of 103 - 106. In our study from road side vendors 12 Salmonella, 24 Aeromonas, 11 Bacillus, 29 Entarobacter spp, 29 E.coli, 30 Staphylococcus aurues 29 Klebsealla spp 5 Aspergillus fumigates, 10 Aspergillus niger and 3 Aspergillus flavus have been identified. In fast food outlets 26 Salmonella, 21 Aeromonas, 9 Bacillus, 30 Entarobacter spp, 30 E.coli, 30 Staphylococcus aurues 30 Klebsealla spp and 9 Aspergillus fumigates and 2 Aspergillus flavus have been identified. In family restaurants 21 Salmonella, 15 Aeromonas, 9 Bacillus, 30 Entarobacter spp, 30 E. coli, 30 Staphylococcus aurues 30 Klebsealla spp 4 Aspergillus fumigates, 3 Aspergillus niger and 4 Aspergillus flavus have been identified. Availability: Items available for loan: UVAS Library [Call number: 1413,T] (1).

3. Microbiological Quality Of Commercial Fruit Juices Sold In Lahore City

by Muhammad Naeem Iqbal | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print Publisher: 2012Dissertation note: Fruit juices are used for their nutritional value, thirst quenching properties and stimulating effect or for their medicinal values. Due to poor hygienic conditions during processing and packaging, fruit juices are becoming a health hazard. Many outbreaks are caused by consuming poor quality juices. Food borne infections are caused by eating food or drinking beverages contaminated with bacteria and parasites. Most of the food borne infections remains undiagnosed and unreported due to poor documentation system and failure in the implementation of law regarding food borne diseases in Pakistan. The present study was conducted to compare the quality of commercial fruit juices so as to provide data for local authorities to deal food security issue. A total of ninety packed fruit juice samples were obtained from retail shops in Lahore city. The fruit juice samples included, apple, mango and orange juices of five various brands. The pH value of the fruit juices measured using pH meter was found between 2.0 to 4.0. Bacterial load of fruit juices was assessed using Total viable count, Staphylococcal count and Coliform count to compare the quality of fruit juices.All the samples were positive for total viable count, 60 samples were positive for staphylococcus count and 30 samples ere positive for coliform count.The mean total viable count in fruit juice samples was3.70xIQ5CFU/ml (log 5.56±1.47CFU/ml)with the range from log 2.69 to log 7.67CFU/ml.Mean staphylococcal count in fruit juice samples was 1.34x 102CFU/ml (log 2.11±1.97CFU/ml) with the range from log 0.00 to log 5.62CFU/ml. Sixty out of 90 fruit juice samples showed staphylococcal counts.Mean coliform counts of 1.80xlOI CFU/ml (log 1.25±1.57CFU/ml) with the range from log 0.00 to log 5.50 CFU/ml. Thirty out of 90 fruit juice samples were positive for coliforms. Identification of bacteria was done on the basis of culture characters, microscopic characters and biochemical tests as per standard protocols described in Manual of Food Microbiology. Among the 226 bacterial isolates, Bacillus spp. were (150),Staphylococcusaureus (49)and E. coli (27), and no Salmonella were detected from the collected samples. Although fruit juices have low pH, still higher viable counts and prevalence of bacterial isolates suggest poor hygienic conditions during manufacturing procedures. Antimicrobial sensitivity profile of the isolates was studied by standard Disk diffusion method (Kirby Bauer method) for commonly used antibiotics. Among various antibiotics used, highest97.78% resistance toAzlocillinand lowest 25.22% resistance against Sulphafurazole. These findings suggest that the antibiotic resistance is transferred through fruit juices. After microbiological examination, it was cleared that fruit juices were as contaminated as compare to our country standards and hygienic conditions. Availability: Items available for loan: UVAS Library [Call number: 1417,T] (1).

4. Physico-Chemical Factors Affection Survival Of Mycoplasma Gallisepticum

by Javed Muhammad | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.

Material type: book Book; Format: print Publisher: 2012Dissertation note: Poultry industry is second largest industry in Pakistan. Mycoplasma gallisepticum causes chronic respiratory disease in poultry and has a great impact on economy of country. The present study was conducted to check the effect of physical factors including pH, temperature, ultra violet light (UV), atmospheric condition and sodium chloride, chemical factors including formalin and sodium hypochlorite and extracts of herbal plants including Garlic, glycyrhiza and Neem on survival of Mycoplasma gallisepticum (MG). Referenced isolate of MG received from University Diagnostic Lab (UDL), University of Veterinary of animal Sciences, Lahore was used in Bacterial count 0.1 at optical density 450 equal to approximate 108 cfu/ml was used in the entire experiment. Survival of MG at pH level 4.8 and 10.8 is significantly lower (p?0.05) as compared to pH level 7.8. Optimum pH was found 7.8 showing best growth while pH 10.8 indicated more lethal effect on survival of MG as compared to 10.8. Temperature study showed that MG exposed to 43°C more lethal effect on survival as compared to 31°C while showed growth occurred at 37°C. Ultra violet light (254nm) showed significant effect (P?0.05) on viability at a distance of 2, 4 and 6 centimeter which indicated that MG at 2 cm from UV light leading to death with increase in exposure time. Survival of MG was best in presence of 5 to 10% CO2 or candle jar as compared to incubate in closed container or without closed container and candle jar (open air). Sodium chloride 3 and 5 percent occupied a drastic effect on MG viability but resistance was existed up to some extant to 1 percent. Culture of MG exposed to formalin 0.1 and 0.2 percent for 15 minutes resulted in high lethal effect significantly (p?0.05) as compared to 0.05 percent. Non significance difference (P?0.05) was present between 4 and 6 percent sodium hypochlorite in terms of effect on survival of MG and has lethal effect when exposed for 5 minute which differ significantly from 2 percent which resulted in death after exposing for 10 minutes. Glycyrhiza and Neem indicated minimum inhibitory effect against MG with similar concentration of 6.25 mg/mL while garlic stop growth at concentration of 3.125 mg/mL. Availability: Items available for loan: UVAS Library [Call number: 1446,T] (1).

5. In Process Quality Control Factors Affecting Potency Of Inactivated Black Quarter Vaccine

by Kashif Hanif | Prof. Dr. Khushi Muhammad | Dr. Aftab Ahmad Anjum | Prof. Dr.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2013Dissertation note: Black quarter (BQ) is a majorbacterial disease of cattle and buffalocharacterized by loss of appetite, lameness, depression, fever, swelling of the skeletal muscles, crepitating sounds followed by sudden death without any clear signs of disease. It results in irrecoverable economic losses. The disease prevails in all provinces of Pakistan especially in District Dera Ismail Khan,Cholistan and Chakwal etc. Morbidity losses comprise of losses due to reduced milk production, work hindrance, treatment charges etc. The survey revealed that 15.91% losses were due to morbidity and 84.09% losses occurred due to mortality caused by black quarter in cattle and buffalo. Reliable diagnosis, mass scale vaccination and clamping strict bio-security measures are the only ways to control the disease.The present study was aimed to optimize the PCR for prompt and reliable diagnosis of BQ and to evaluate the inactivated whole culture vaccine with variable biological titer to induce protective immune response in calves. The comparative antibody response of animals to adjuvanted (Aluminium hydroxide gel & Montanide ISA 70) and non adjuvanted 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. All of the vaccines were inoculated in a group of four animals. Serum samples were collected at specified time intervals and antibody levels were detected through antihemolytic units. The PCR was optimized for diagnosis of C. chauvoei in clinical specimens from infected carcasses. Study of factors (temperature, media composition, pH, incubation time and anaerobic agents)for biomass production of bacteria and its hemolytic toxins revealed that certain growth parameters can be improved to enhance the bacterial growth and its hemolytic toxins. Like use of RCM medium for vaccine production enhances the growth of C. chauvoei and its toxins under in vitro conditions. Supplementation of nitrogen gas in culture medium can enhance the bacterial growth and hemolysin. Proper incubation time, temperature and pH can be very helpful factors for the growth and biomass production of C. chauvoei under in vitro conditions. Finally, biomass production of the organism using manual biofermentor is a very cheap and cost effective method for concentrated vaccine production in our country where commercial biofermentor cannot be afforded. So by using these techniques we can make more no. of vaccine doses from less quantity of bacterial culture. It will also help us in developing bivalent, trivalent or multivalent vaccine. Study of in process quality control factors (bacterial biomass and toxins) production and "in process quality control" factors (biological titer, bacterial count, hemolytic units, adjuvants and storage time) that affect antibody response of vaccinatesrevealed that vaccine with 250 HU / dose showed relatively similar antibody titer in calves as the vaccine with 500 HU or 750 HU per dose. So this can be helpful to produce more doses of vaccine with same culture. The Montanide ISA 70 gave best result for development of good and prolonged immunity but gel based vaccine also produce satisfactory results.So in future oil based vaccine may be used to attain long term and effective immunity. Effect of priming and boosting revealed that boosting give better results as compared to primed group by producing prolonged immunity. The results were very encouraging. Effect of storage showed that the quality of immunogen was not affectedwith the passage of time if the vaccine is properly stored at 4 °C upto three months. Availability: Items available for loan: UVAS Library [Call number: 1586,T] (1).

6. 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.

Material type: book Book; Format: print ; Literary form: not fiction 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. Availability: Items available for loan: UVAS Library [Call number: 1732,T] (1).

7. 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: book Book; 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).

8. Antibody Response Of Goats To Bivalent Pprv And Goat Pox Virus Vaccine

by Muhammad Farooq (2009-VA-146) | Prof. Dr. Khushi Muhammad | Dr. Muhammad Anees | Professor Dr. Aftab Ahmad Anjum | Dr. Muhammad Avais.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: PPR is one of the most acute viral disease of sheep, goats, deer and other similar animals. This disease is caused by Morbillivirus which belongs to familyParamyxoviridae. In this disease oculo-nasal discharge, stomatitis, diarrhea, high temperature, and pneumonia is common sign.along with foul breath.Goat pox virus (GPV) disease is extremely transmissible described by temperature, falling down and different stages of pox lesion development such as, vesicles, scars, pustules, erythema and papules, all over the body. This study was aimed to evaluate the monovalent lyophilized PPRV and GPV vaccine with bivalent PPRV and GPV vaccines. Moreover effect of amount of immunogen of the vaccines, and nature of adjuvant used in the vaccine on antibody response of goats was also evaluated.Seven types of vaccines PPR (FD), GPV (FD),PPR+GPV (FD), PPR+GPV(gel 102.5), PPR+GPV(gel 103.5), PPR+GPV(gel 104.5) and PPR+GPV(oil) were prepared. All vaccines other than gel based contained 103.5 immunogen level. Each vaccine was inoculated to each of the six goats of the respective group. Blood was collected at 21, 42 and 63 dayspost vaccination. The antibody response of goats was measured with CFT. There was non-significant difference between the anti-PPR antibodies induced by either monovalent or bivalent vaccines. Similarly goat Pox vaccines also produced non-significant difference in both monovalent and bivalent form. Antibody response was directly proportional to the amount of specific immunogen in the vaccine. There was non-significance effect of gel or oil in the vaccine as an adjuvant on the antibody response of goats to the vaccine. Availability: Items available for loan: UVAS Library [Call number: 2496-T] (1).

9. Spatial Ecology And Distribution Of Soil Borne Burkholderia Mallei In Punjab, Pakistan

by Muhammad Asad Ali (2002-VA-73) | Prof. Dr. Khushi Muhammad | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Mansur-Ud-Din Ahmad.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Burkholderia mallei is a causative agent of glanders, the disease of equines. The disease is characterized by pulmonary, nasal and cutaneous forms. B. mallei is excreted through nasal discharge, lacerated skin/wounds and expiration. Diseased animals shed bacteria through the discharges contaminating soil, water, fodder and other susceptible animals in its vicinity. The present study was designed to map and investigate the association of different physical factors and soil chemistry analytes with persistence of B. mallei genome in soil of 10% percent villages (n=456) from eight selected districts of Punjab province, Pakistan. Eleven (0.48%) out of 2, 280 soil samples were positive for B. mallei genome in varied locations of Punjab. Higher prevalence (2.37%) for genome was detected in Sheikhupura district followed by Chakwal district (2.10%). None of the samples from Gujranwala, Sahiwal, DG Khan, Attock, Faisalabad and Sargodha districts were found positive for B. mallei genome. The genome of B. mallei was distributed in 25% study districts of Punjab, Pakistan. In Chakwal district, the genome of B. mallei was strongly associated with moisture (p=0.008) in all positive samples ranging from 0.80 to 39.20%, Phosphorous (p=0.050) ranging from 1.74 to 21.75 mg/Kg. While, this association in Sheikhupura district soil samples was with Sodium (p=0.018) and moisture (0.026) ranging from 1.90 to 133.59 mg/Kg and 0.80 to 39.20%, respectively. The odds of detecting DNA of B. mallei were recorded higher (1.4, 6.8, 5.0, 2.8 and 10.6 ) when soil sample sites were < 500 meters away from vehicular traffic roads, < one kilometer from animal markets, < 100 meters from canal, animal density < 1,000 animals and human population < 300 houses/village. While the odds of detecting DNA of B. mallei were 0.1, 0.3, 0.4, 0.2 and 0.5 when soil sample sites were > 500 meters from vehicular traffic roads, > one kilometer from animal markets, > 100 meters from canal, animal density > 1000 animals and human population > 300 houses/village, respectively. Soil-borne B. mallei DNA is more likely to be detected in areas closer to roads with vehicular traffic along the interstate routes in Punjab and soil containing low level of moisture. It was concluded that soil of two districts out of eight selected was positive for B. mallei genome in Punjab province. Odds of less distance from main road to animal farm and high animal density at farm were positively associated with B. mallei DNA persistence in soil. Moisture, sodium and phosphorus were positively associated with persistence of B. mallei DNA in soil. Availability: Items available for loan: UVAS Library [Call number: 2900-T] (1).

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