1.
Food Borne Bacterial Contamination Of Retail Poultry Meat At Chicken Sale Outlets In Lahore City
by Anwar Ullah | Dr. Aftab Ahmad Anjum | Dr. M. Younus | Dr. Tahir yaqoob.
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Publisher: 2010Dissertation note: Poultry is an important sub sector of live stock and its share in GDP is 2.0 percent and it is playing an important role in improving the income of rural and urban population of Pakistan. Food poisoning is becoming a serious health problem to human being. The organisms belonging to the Genera Salmonella and Campylobacter are considered to be the most important pathogens. The present research is performed for isolation and enumeration of bacterial pathogens from the family Enterobacteriaceae and Genus Staphylococci.
A total of 200 samples each of liver, kidneys, breast muscle and intestine were collected from four different roads of Lahore city. Each sample was weighed 5gm and was rinsed in 10 ml sterile phosphate buffer saline (PBS) for collection of Sample rinses. Serial 10-fold dilutions of sample rinses (SR) were subsequently prepared in sterile phosphate buffer saline (PBS) for standard plate count (SPC). Each dilution of sample rinsed were inoculated to selective agar plates for enumeration and isolation S. enteritidis, C. jejuni, E. coli and S. aureus.
Numbers of selective agar plates contaminated with E. coli were 75.5%, having the highest value and S. aureus were isolated from the lowest number of plates (50%). C. jejuni were isolated from 119 (59.5%) out of 200 samples and Salmonella enteritidis contamination rate were 71% (142 out of 200 samples). Over all 200 samples each of liver, kidneys, breast muscles and intestine 158(79%) out of 200 were contaminated with food borne bacteria. Kidney samples were less contaminated having contamination rate of 42.5% (85 out of 200 samples). Intestine and liver samples were contaminated with a medium range of 66.5% (133) and 68% (136) respectively.
Intestinal samples had highest range of colony forming units (CFU) per gram of C. jejuni and kidney samples had the lowest CFU per gram with an average CFU per gram of 1.2 x106 (<log 6.09>) and 1.0 xi (<log 4.02>)respectively. Liver samples had an average CFU per gram of 6.3 xl ü (<log 4.80>) and breast muscle had 1.4 x ] (<log 5.15>)CFU per gram of sample(Table 02). Liver samples had highest CFU per gram of S. enteritidis and kidney samples had the lowest value with an average CFU per gram of 1.1 x106 (<log 6.05>) and 9.8 xi0 (<log 3.99>) respectively. Intestinal samples had an average 9.7 xiO5 (<log 5.98>) and breast muscle had 1.1 xi (<log 5.04>) CFU per gram of sample (Table 03). Intestinal samples had highest CFU per gram of E. coli and kidney samples had the lowest value with an average CFU per gram of 7.7 x l0 (<log 7.88>) and 1 .5 x (<log 4.19>) respectively. Liver samples had an average 8.7 x 0 (<log 5.94>) and breast muscle had 1.8 x 106 (<log 6.25>) CFU per gram of sample (Table 04). The breast muscles had the highest CFU per gram of S. aureus and kidney samples had the lowest value with an average CFU per gram of 1.1>i0 (<log 5.04>) and l.5x 10 (<log 4.17>) respectively. Liver samples had an average of 1.1 xl (<log 5.05>) and intestinal samples had 2.5x10 (<log 4.40>) CFU per gram of samples.
The samples were sub cultured for isolation and identification of food borne pathogens like Salmonella enteritidis, Campylobacter jejuni, Escherichia coil and Staphylococcus aureus species by following the standard protocols of Bergey's Manual of Determinative Bacteriology 9th Edition.
Availability: Items available for loan: UVAS Library [Call number: 1185,T] (1).
2.
Characterization Of Indigenious Species Of Mycotoxins Producing Aspergilli
by Gull Naz | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.
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Publisher: 2010Dissertation note: Pakistan's economy is based on agriculture. Agriculture crops are harvested and stored in feed mills for production of thousands ton of feed for livestock as well as poultry through out the year. In Pakistan, July and August are hot and humid months during which moulds grow abundantly on the heaves of wheat! rice/maize straw and feed ingredients and produce variety of toxins.
Present study has been designed to explore different groups of moulds prevailing in and around Lahore city in each month of the year. Samples of soil and air were collected from ten different places of Lahore city.
A total of 240 samples were cultured on a common Saboraud's Dextrose Agar to get single colonies of each mould. These single colonies were identified by colony characters, slide cultures and biochemical tests. Mycotoxin producing Aspergilli were isolated by culturing on specified media and placing the cultures under Wood's lamp. Mycotoxin productions potential were assessed by extracting mycotoxins of these Aspergilli. Mycotoxins produced by the Aspergilli were identified and purified through Thin Layer Chromatography. These mycotoxins were then quantified through High Performance Liquid Chromatography.
The identified and purified mycotoxins can be used as standards. Reference standards are important and critical for qualitative and quantitative detection of mycotoxins in field samples screening. Presently mycotoxin is a ban item. The occurrence of toxinogenic Aspergilli have economic impact directly on livestock and poultry products export.
Availability: Items available for loan: UVAS Library [Call number: 1217,T] (1).
3.
Detection Of Hazardous Organism In Raw And Pasteurized Milk With Particular Reference To 3Enterobacteriaceae
by Ayesha | Prof. Dr. Mansur ud Din Ahmad | Dr. Aftab Ahmad Anjum | Prof. Dr.
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; Literary form:
not fiction
Publisher: 2010Dissertation note: The present study was carried out to detect the hazardous organisms in raw milk from public health point of view. In total one hundred twenty (120) milk samples were collected from milk retail shops in and around Lahore. Out of these 120, one hundred samples were of raw milk and rests of the twenty samples were of pasteurized milk. Their microbiological quality was studied by performing standard plate count (SPC), coliform count and identification of hazardous bacteria belonging to the family Enterobacteriaceae. The micro flora of milk was also studied for the prevalence of multiple drug resistant (MDR) bacteria.
Milk supplied in Lahore city was found to have poor microbiological quality. Bacterial load was determined by SPC and coliform count. The standard plate count (S.P.C) of the raw milk ranged from 4.2x106 to 7.7xl07 c.f.u/ml. The coliform counts ranged from 3.4x 104 c.f.u /ml to 6.9x105 /ml. A total of 81 isolates were identified from raw milk samples. These included Yersinia (3 strains), Klebsiella (16 strains), Escherichia coli (14 strains), Enterobacter (11 strains), Shigella (3 strains), Salmonella (19 strains) and' Proteus (15 strains).The standard plate count for pasteurized milk ranged from 1.45x104 c.f.u/ml to 3.8x 105 c.f.u/ml. The minimum and maximum coliform count was 7.2x102 to 8.4xl03 c.f.u/ml respectively for pasteurized. All samples were outside the international standard for coliform bacteria. A total of 13 isolates were identified from pasteurized milk samples. These included Yersinia (2 strains), Klebsiella (1 strains), Escherichia coli (6 strains), Enterobacter (2 strains), Shigella (1 strains) and Proteus (1 strains).
All the isolates showed multiple drug resistance to various commonly used antibiotics in veterinary practices. Escherichia coli were resistant to all antibiotics used except Gentamicin (10µg). Enterobacter was sensitive to all the antibiotics used except to Ampicillin (10µg). Shigella was sensitive to Gentamicin (10µg), Kanamycin (30µg), Choloramphenicol( 25µg), but showed resistance to Ampicillin (10µg), Oxytetracycline ( 25µg), Streptomycin (10 µg), Pencillin (10 µg) and Tribrissin (25µg)., Salmonella was resistanct to Ampicillin (10µg), Oxytetracycline ( 25µg), Streptomycin (10 µg), Pencillin (10 µg) and Tribrissin (25µg). But sensitive to Gentamicin (10µg). .All the isolates showed greatest resistance to Penicillin (10 ug.) whereas, most of the isolates were sensitive to Gentamycin, Kanamycin and Chloramphenicol.
Finally, it is recommended that the members of the public should always boil raw milk before consumption because of their microbial content. Therefore, it is highly recommended that hygienic practices and regulations, such as on-site pasteurization and implementation of HACCP following established standards, should be introduced to facilitate the production of raw milk of high quality and safety.
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4.
Physico-Chemical Growth Requirements And Molecular Characterization Of Indigenous Spirulina
by Muhammad Qasim | Dr. Imran Najeeb | Dr | Dr. Aftab Ahmad Anjum.
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Publisher: 2010Dissertation note: Spirulina is a microscopic and filamentous cyanobacterium (blue-green alga). It is 60-70% protein by weight and contains a rich source of vitamins, especially vitamin B12 and provitamin A (13-carotene), and minerals, especially iron. One of the few sources of dietary y-linolenic acid (GLA), it also contains a host of other phytochemicals that have otential health benefits. For medical scientists it is gaining more attention as a nutraceutical and source of potential pharmaceuticals. Spirulina has ability to inhibit viral replication, strengthen both the cellular and humoral immunity and cause regression and inhibition of cancers it also has antioxidant property. It also has been receiving increasing interest due to its potential to produce a diverse range of chemicals and biologically active compounds, such as vitamins, carotenoid pigments, proteins, lipids and polysaccharides.
Present study was designed to explore the indigenous spirulina and its mass cultivation by optimizing the physicochemical growth requirements. One hundred and twenty samples were collected from different soils and water reservoirs from three districts (Sargodha, Lahore and Faisalabad) of Punjab. Then spirulina was isolated from collected samples and cultivated under different nutrient, temperature and light regimes to get its maximum bio-mass in our laboratory.
Our results showed that maximum growth of indigenous spirulina was obtained at 30°C and at 1500 lux (light intensity). Nitrogen concentrations (0.625. 1.25 and 1.875 gIl) had no effect on the growth, while phosphate concentrations (0.5, 1.0 and 1.5 gIl) had a minimal and gradual effect on growth as the concentrations were increased. For the confirmation and molecular characterization of indigenous spirulina, DNA was isolated by chioroform-isoamyl alcohol extraction method and its polymerase chain reaction (PCR) was carried out by using specific primer of 16s rDNA gene (CYA1O6F and CYA78IR) and PCR products were run on gel giving an amplicon size of 700 bp.
Now a day in the world people are competing for food supplementation. The spirulina can act as a source of nutraceuticals. This study helps in optimizing the growth of indigenous spirulina. For large scale industrial production its extensive study should be done like physiology, growth, reproduction etc. This will pave an avenue for further nutraceuticals.
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5.
Prevalence Of Multiple Drug Resistant (Mdr) Bacteria In Intestinal Infections Of Dogs
by Iffat Habib | Dr. Aftab Ahmad Anjum | Prof. Dr. Masood Rabbani.
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Publisher: 2011Dissertation note: Antimicrobial resistance is a complex problem involving various bacterial species, resistance mechanisms, transfer mechanisms and reservoirs. Cats and dogs are the potential sources for spread of antimicrobial resistance in humans due to their close contact with them. The horizontal transfer of antimicrobial resistance genes through plasmids, integrons and transposons has been found to play an important role in the dissemination of antimicrobial resistance genes. Canine antimicrobial resistant genes had been identified in bacteria isolated from human clinical infections suggesting the spread of resistance mechanisms from canine to human bacteria.
The present study has been designed to study the prevalence of multiple drug resistant strains causing enteritis in dogs. 100 Samples were collected from different Pet clinics in and around of Lahore city. These samples were cultured for identification of MDR bacteria. Antibiotic resistance profile was studied by the standard Disk diffusion method (Kirby-Bauer Method) for commonly used antibiotics. These MDR bacteria were isolated and identified as per standard protocols described in the Bergey's Manual of Determinative Bacteriology. Different combinations of antibiotics were also evaluated for in-vitro antibiotic sensitivity for an effective treatment of these cases so that the load of MDR bacteria could be reduced.
From the collected samples E. coli, Salmonella enterica, Proteus vulgaris, Citrobacter diversus and Psedomonas spp. were identified. Among all of these E.coli was most prevalent followed by Salmonella enterica, Citrobacter diversus, Proteus vulgaris and Psedomonas spp. Out of 127 E.coli isolates 52 40.94%) were declared as MDR-Bacteria following 50 Salmonella enterica isolates 17 (34.00%), 17 Citrobacter diversus 6 (35.29), 12 Proteus vulgaris isolates 06 (50%). It was concluded that MDR isolates were most sensitive to antibiotic combination (Amoxicillin + Clavulanic Acid), followed by (Oxytetracyclin + Tylosin), (Gentamycin + Ceftriaxone), and (Penicillin + Streptomycin). Out of 52 MDR E.coli isolates 23 (44.23%) were found to be invasive. Recommendations are made on prudent use of antimicrobial drugs in dogs, as well as on the need to develop science-based infection control programs in veterinary hospitals.
Availability: Items available for loan: UVAS Library [Call number: 1231,T] (1).
6.
Preparatuin And Evaluation Of Monospecific Anisera Against Hemagglutinin, Neuraminidase And Matrix Proteins of local Avian Influenza Strains H5 N1, H7 N3, H9 N2, for diagnostics
by Sumaira Ijaz | Dr. Atif Hanif | Dr. Aftab Ahmad Anjum | Prof. Dr.
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Publisher: 2011Dissertation note: Avian Infuenza is an economically important disease of poultry worldwide. It has caused losses to poultry industry on larger scale. It is important due to its zoonotic nature. Studies were carried out to raise monospecific antisera against hemagglutinin, neuraminidase and matrix antigens. HA, NA and M proteins of each of the avian influenza strains were separated on Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis (SDS PAGE). First virus was lysed to release the proteins. Virus was lysed by using 4% Triton X 100, 1mM KCl and 0.01M Tris buffer. Then the sample was dialyzed. Sample was run on gel to purify proteins. The protein bands of appropriate molecular weight were cut and triturated in 1ml of normal saline. Material was centrifuged to remove the gel content.
Each protein was confirmed by the Bradford's Reagent. Each protein was individually mixed with Montanide ISA 50 adjuvant in 1:1 ratio to make the vaccine. Vaccine of each polypeptide of AIV strains was injected in three groups of nine birds each. One group of birds was injected with HA, second group with NA and third group with Matrix proteins of H?, H? and H?. Three groups of birds served as control. The blood samples of all birds were collected before and after inoculating vaccine. The sera of birds before and after inoculating vaccine were checked for antibodies titre against HA antigen by HI test. Antibodies against Matrix antigen were detected by Agar Gel Precipitation Test. Antibodies titre was raised after inoculating polypeptides. In case of sudden outbreaks, antisera may be helpful to control disease.
Availability: Items available for loan: UVAS Library [Call number: 1351,T] (1).
7.
Monitoring Of Mixed Vegetable Salads For Microbial Quality
by Sana Hameed | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.
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; Nature of contents: ; 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).
8.
Microbiological Quality Of Commercial Fruit Juices Sold In Lahore City
by Muhammad Naeem Iqbal | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.
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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).
9.
Clincal Cytogenetic Investications In Cattle & Buffalo Population Of Punjab For Chromosomal Abnormalities
by Muhammad Bilal Bin Majeed | Prof. Dr. Khalid Javed | Dr. Aftab Ahmad Anjum | Dr. Ahmad Ali.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1439,T] (1).
10.
Physico-Chemical Factors Affection Survival Of Mycoplasma Gallisepticum
by Javed Muhammad | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.
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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.
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11.
Efficacy Of Commercial Disinfectants Against The Water Contaminating Bacteria At Commercial Broiler Farms
by Mian Muhammad Salman | Dr. Aftab Ahmad Anjum | Pfor. Dr | Prof. Dr. Tahir Yaqub.
Material type: ; Format:
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Publisher: 2012Dissertation note: Water is an important constituent for poultry. Poor hygienic conditions of water are health hazard for poultry. Many outbreaks are caused by consuming poor quality water.. Fifty water samples from different broiler farms in and around Lahore were collected from drinkers in sterile containers. Bacterial load was assessed using total viable count and coliform count. The counts were above the threshold level (50cfu/ml for coliform and 100cfu/ml for total viable count) showing that water used at poultry farms was of low microbiological quality. Five Disinfectants PHMB20% (.75ml/lit, 1.5ml/lit, 3.0ml/lit), PHMB11% (1.5ml/lit, 3.0ml/lit, 6ml/lit), 0.2% chlorine dioxide (0.1ml/lit , 0.2ml/lit, 0.4ml/lit) Glutral 9.8%(1.5ml/lit, 3.0ml/lit, 6ml/lit), organic acid(1.5ml/lit, 3.0ml/lit, 6ml/lit) were used and they resulted in log reduction of TVC by PHMB20% (5.83±4.36, 6.14±3.98, 9.35± 0.68), PHMB11% (9.42±0.21), 0.2% chlorine dioxide (2.45±0.97, 3.19±0.73, 6.33±0.80 ) Glutral 9.8%(6.87±1.00, 9.73±1.00,9.73±1.00), organic acid(4.75±1.21, 6.62±1.26, 6.90±1.15).PHMB20%,PHMB11%, Glutral 9.8% and organic acid were effective at normal dose, while 0.25 chlorine dioxide was effective at normal dose against at normal dose. Log reduction in Coliform count at half, normal and double dose by PHMB20% (6.52±3.33, 6.96±2.46, 7.96±0.98), PHMB11% (7.89±1.01), 0.2% chlorine dioxide (3.65±0.73, 5.08±0.98, 6.27±0.97) Glutral 9.8%(8.48±0.99), organic acid(5.18±1.21, 5.93±1.26,6.46±1.15±) . PHMB20%, PHMB11%, 9.8% Glutral, organic acid were effective against coliform bacteria at half dose while 0.2% chlorine dioxide was effective at normal dose. Glutraldehyde was effective at normal dose amongst all disinfectants against Total viable bacteria and coli form bacteria.
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12.
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: ; Format:
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; 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).
13.
Prevalence Of Salmonella And Campylobacter Contamination In Poultry Eggs
by Hassaan Bin Aslam | Dr. Aftab Ahmad Anjum | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Provision of adequate food to their inhabitants and assure an atmosphere free from hunger and malnutrition is the responsibility of a civilized government. The food security objective becomes more important when 15-20% of the world population is not getting sufficient food to meet minimum nutritional requirements for a healthy and productive life. Proteins play an important role in the formation of balanced human diet. There are mainly two sources of proteins i.e. animals and plants. Commercial egg production is an important economic enterprise offering more rapid and efficient return than many other livestock production operations. In 1999-2000, 13.9 million commercial layers in Pakistan produced 3,261 million eggs contributing 27% eggs to the total egg production of 8677 million eggs.
The incidence of food borne diseases is increasing globally. Many cases of food borne illness occur as a result of improper food handling and preparation by consumers in their own kitchens. Some of the most compelling evidences have come from the international data on Salmonella and Campylobacter species infections. Food-producing animals (e.g., cattle, chickens and turkeys) are the major reservoirs for many of these organisms. A total of 500 raw chicken eggs were bought from different retail outlets in Lahore city for determining the prevalence of Salmonella and Campylobacter inside and on the egg shell and enumeration of their load. Samples were graded as clean, trace, dirty and cracked. All samples were processed inside the safety cabinet. Eggs were broken from narrow end and contents were drained in the petri plate. Egg shell and egg shell membrane was processed by shell crush method in a tube in the presence of phosphate buffer saline. Sample from egg yolk and albumin was taken by direct aspiration while egg shell rinse was taken as a sample for Salmonella and Campylobacter. Isolation is done by enrichment method for this purpose selenite broth and buffered peptone water is used for Salmonella and campylobacter, respectively. The enriched sample was then plated on the Brilliant Green Plate selective for Salmonella and Campy Cefex Agar plate that is selective for Campylobacter. Thirteen samples out of 500 hundred samples were positive for presence Salmonella with over all prevalence of 2.6%. Highest percent prevalence was found in cracked eggs where it is 33.3% followed by dirty and clean eggs (0.9%) and trace eggs with zero prevalence. Colony forming units of Salmonella on the shell of one positive sample is 4.2x102 while CFUs in egg yolk of cracked egg was 7.0x102. Regarding Campylobacter five eggs out of 500 eggs were positive with overall prevalence of 1%. The highest prevalence was found in cracked eggs where it was 16.6% and dirty eggs having prevalence of 12.5%. Both clean and trace eggs were having zero prevalence. CFUs for Campylobacter were too low to count while majority of samples were observed negative for viable CFUs for Campylobacter.
Availability: Items available for loan: UVAS Library [Call number: 1614,T] (1).
14.
Isolation And Characterization Of Phytase Producing Microrganism From Soil
by Ghazal Aziz | Dr. Aftab Ahmad Anjum | Prof. Dr | Prof. Dr. Tahir Yaqub.
Material type: ; Format:
print
; Literary form:
not fiction
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.
Availability: Items available for loan: UVAS Library [Call number: 1685,T] (1).
15.
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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16.
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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17.
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.
Availability: Items available for loan: UVAS Library [Call number: 1732,T] (1).
18.
Immunohistochemical Identification Of Adenovirus Type 4 In Liver, Heart, Kidney, And Pancreas Of Broiler Chicken
by Muhammad Tanzil-ur-Rehman | Dr. Muti-ur-Rehman Khan | Dr. Aftab Ahmad Anjum | Dr. Yasin Tipu.
Material type: ; Format:
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drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1736,T] (1).
19.
The Development Of Tea Whitener By Partial Replacement Of Palm Oil With Canola Oil
by Junaid Kabir | Dr. Muhammad Nasir | Dr. Aftab Ahmad anjum | Dr. Saima.
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Publisher: 2013Dissertation note: Tea whitener is now become a popular trend in Pakistan with 01 billion tons consumption annually according to my personal information and its consumption is increasing day by day. The replacement of hydrogenated palm oil used traditionally is necessary as they contains 49.3% saturated fatty acids, the majority of which are palmitic acid, myristic acid and lauric acid which are proved to be most injurious for human health, raises the total and LDL cholesterol (Bonanome et. Al, 1998). Canola oil is known for its low level of saturated fatty acids, a relatively high level of monounsaturated fatty acids, and a very good amount of the n-3 fatty acid a-linolenic acid. Canola oil consists of an appreciable amount of a-linolenic acid which amounts for almost 10 percent which is a fairly good quantity. In addition, 1:2 is the ratio balance between linolenicacid and linoleic acid which is favorable and well balanced. Canola oil is a relatively rich source of tocopherols,60-70 mg/100g, contains high level of phytosterols (892 mg/100 g. Keeping in mind the above mentioned nutritional aspects, canola oil based tea whitener is developed.
The research was conducted in two phases. During 1stphase the HLB requirement of the canola oil and partially hydrogenated palm oil was determined which are determined as 08 and 06 respectively. Then the emulsifier's percentages are calculated according to their standard HLB values and the doses of the emulsifiers "DATEM" and "GMS" are adjusted according to the ratios of hydrogenated palm oil and canola oil in all the formulations. During second phase the proximate, chemical, physical and sensory analysis are done for all the emulsions so as to determine their resemblance with the control formulation S1.
Different graph analysis regarding proximate analysis of canola oil based tea whitener showed that the results for moisture percentage are (85.69 ± 0.089), for crude protein the values are (1.66±0.22), for dry matter its (14.32±0.04) for crude fat the result shows (7.01±0.03). The variation in all the emulsions were negligible, as the ingredients except fat source is almost the same in all the formulations.
The results for acidity of tea whitener emulsions on the 2nd day which is 0.09± 0.02 which shows the acceptable range, while on the 6th day the mean value of the acidity is also in the acceptable range which is 0.14± 0.01 which means acidity increases to some extent on the 6th day of storage. The mean of the acidity on the 8th day is 0.16± 0.01. The trend shows the acidity increases from 0.09±0.02 to 0.16±0.01 in 08 days. The variation was observed in all the emulsions with the passage of time, but there is not a very significant difference among all the emulsions as compared to control S1.
Mean values for pH on the 2nd day is 6.79±0.03 while the control sample S1 has the pH value of 6.82 on the 2nd day and the treatment which has the lowest pH values on day 2nd is S6 with pH 6.75. The observations on 6th days are shown in pH chart which shows slight decrease in pH in the 6th day with the mean value 6.71±0.02. The mean value of pH on the 8th day is 6.61±0.02. The results showed that pH of tea whitener emulsionsdecreases as storage progressed. A very interesting point raised during study that the pH values of the standard emulsion S1 is higher among all the emulsions on the 2nd day, but as the days proceeds, the pH of the emulsions with different rations of canola oil retains their pH and the pH becomes almost the same as standard on the 8th day. This may concludes that the emulsions containing canola oil retains their ph more as compared to palm oil based emulsion.
The mean value of density of the tea whitener emulsions 1.12±0.02. So overall the results variation is not significant. The little difference may be due to the fact that palm oil has density of 0.89 L/kg at 25 C while the density if canola oil is 0.91 L/kg on the same temperature. The density of all the formulations are comparable with the control emulsion S1.
The results depicted that 'L" value was decreaseswith increase in the ratio of canola oil. Mean comparison for color "L" parameter showed that highest value for S1 which is 90.45 and least value for S8 which is 89.29. The variation is very slight but the palm oil based emulsions are slight whiter in the appearance.The mean value of a* is -0.285 ± 0.095 which shows a very little variation. The level of greenness decreases slightly as the ratio of the canola oil increases from S1 to S8. The degree of yellowness in the emulsion increases as the ratio of canola oil in increases in the emulsions. The mean value of b* is 2.94±0.27 which shows a slight variation as we go from S1 to S8.
The sensory attributes scores obtained from sensory evaluation by trained panelists varies a lot. Addition of canola oil in place of palm oil significantly alters the flavor, After taste and over all acceptability of the tea made with tea whitener emulsions from S1 to S8, the scores are almost the same up to S4 as compared to control formulation S1 for all the attributes mentioned above. Score decreases from S5 to S8 which is definitely due to the addition of canola oil in the formulations. The sensory attributes like fat separation and color get the same scored almost for all the formulations.
Flavor scores are almost the same up to S5 but the scores decreases significantly from S6 to S8, for the sensory attribute of "after taste" the formulations from S1 (standard) to S4 get good scores means the after taste if the S2, S3 and S4 are comparable to the control emulsion S1 while S5 to S8 get lower scores, For "overall acceptability" S2, S3 and S4 are nearly equivalent and good scores as compared to control formulation with 100 percent palm oil formulation with the mean value of 90±02 which gives a green signal that we can partially replace hydrogenated palm oil with canola oil. The formulation S5 get a little lower score as compared to control one. The formulations from S6 to S8 get lower scores in overall acceptability.
Finally it is concluded that the formulation S4 is the one which can be replaced with the control emulsion S1 for making of tea which means 42.5 percent of the total fat in tea whitener can be replaced successfully with canola oil without compromising the physical, chemical and sensory properties of the tea.
Recommendations
The main aim of this project was to make a tea whitener which is based on healthier and heart friendly oil (canola oil) instead of palm oil. Canola oil has been used as a cooking oil and also in nutritional products like "Ensure Plus" and "Glucerna" due to its health friendly composition. The idea is drawn from the nutritional products compositions whose fat part is mostly consists of canola oil.
In Pakistan, keeping in mind a very huge consumption of tea whitener of 01 billion annually according to my personal information. The production may be much higher as my information may be limited. Keeping in mind the annual production or consumption of liquid tea whitener in Pakistan, the delivery of more healthy oil to the consumers by incorporating it in the liquid tea whitener product seems to be a pretty good idea. It is not only the matter of incorporation of healthy canola oil but also the matter of replacement of saturated fatty acids rich palm oil. Keeping in mind the chemical, physical and sensory properties of tea whitener emulsions S4 with 42 percent canola oil of the oil phase gives similar physical, chemical and sensory properties when compared to control formulation tea whitener S1 with 100 percent palm oil as oil phase. Keeping in mind the composition of the canola oil, if tea whitener is made with 07 percent fat level, in case of S4 (The formulation with resemblance to control up to maximum canola oil extent) canola oil percentage if the total fat is 42.5 percent of the total fat, it will give 0.7 grams of omega-3 as ALA per 250 ml of the tea whitener which means that it will provide 2.8 grams of omega-3 per liter of tea whiteners which can help us to meet up to some extent the ADA recommendations which is 1.3 to 2.9 grams based on 2000 Kcal diet (ADA, 2007)
The real challenge in the making of tea whitener formulations with different ratios of canola oil and palm oil is to make a successful emulsions without fat separation, thanks to HLB system for successful making of emulsions. Another challenge is to mask the after taste of the canola oil which can be prominent in the tea whitener, the after taste of canola oil is masked by milk flavor due to which the successful replacement of palm oil with canola oil up to 42 percent becomes possible. The purpose of the product development of making it a source of omega-3 was successfully met as the results shows the partial replacement of palm oil with canola oil is possible.
From the present study it can be concluded that canola oil can be incorporated in liquid tea whitener up to the percentage of 42.5 percent of the total fat without any persistent change in chemical, physical and sensory properties of the tea whitener. The concluded value of omega-3 which it gives per 250 ml is 0.7 grams according to fatty acid profile given by ADA reports (ADA, 2007). They do not affect the taste or texture of the product. My study showed that the replacement of palm oil with canola oil up to 42.5 percent in tea whitener formulation was acceptable among consumers and also the tea whitener retained its quality and sensory properties after storage for 08 days at 04 C. The tea prepared from S4 has the same sensory properties as the tea made with the control formulation. It is recommended that canola oil based tea whitener should be a introduced in the market for creating awareness among the general population about the role of omega-3 n human health and threats of consuming saturated fatty acids. There are need of studies forefficacy of developed tea whitener whether it beneficially transmit the omega-3 to human body or not and what are the health benefits among the subjects. More research work is required to testify the product under UHT treatment to find out what are the changes in physical and chemical properties of the product up to 03 months, it's emulsion stability and it's sensory properties during and after 03 months of shelf life in tetra packaging.
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20.
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%).
Availability: Items available for loan: UVAS Library [Call number: 1767,T] (1).
21.
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.
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not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1852,T] (1).
22.
Isolation And Molecular Characteracterization Of Staphylococcus Aureus From Raw Milk
by Ibrar hussain | Prof. Dr. Muhammad ayaz | Dr. Imran javed | Prof. Dr. Aftab ahmad anjum.
Material type: ; Format:
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drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1853,T] (1).
23.
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).
24.
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:
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1923,T] (1).
25.
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.
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1991,T] (1).
26.
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:
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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).
27.
Status Of Awareness Among Zoo Workers About Zoonotic Diseases
by Tahir Khan (2012-VA-806) | Prof. Dr. Mansur Ud Din Ahmed | Shelly Saima Yaqub | Dr. Shakera Sadiq Gill | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
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Publisher: 2014Dissertation note: A zoo is a place where wild animals are kept for exhibition purposes to the public.It includes: aquaria, sanctuaries, bird gardens and safari/wildlife parks. These are centers for wild animal’sconservation and for public recreation and education (Cuaron2005). Epidemiologists, wildlife biologists, veterinarians and conservationists used these for research purpose.According to an estimate Pakistan is maintainingapproximately 27 zoos, deer parks, etc.(Walker 2014).
Zoonotic diseases are those which are naturally transmitted from animals to human beings and vice versa. The word Zoonosis is derived from the Greek word zoon (animal) and nosos (disease). The diseases which are transferred from human beings to animals are known as Zooanthroponotic (Greek “Zoon” = animal, “anthrópos” = man, “nosos” = disease) diseases e.g. tuberculosis, measles, giardiasis and amoebiasis. On the other hand the diseases which are transmitted from animals to human beings are known as anthropozoonotic diseases e.g. anthrax, AIDS, psittacosis and rabies (Epstein and Price 2009).
Zoonosis can be classified according to their circulation in the ecosystem. These are either classified as synanthropic zoonosis, with an urban (domestic) cycle in which the source of infection are domestic and synanthropic animals (e.g. cat scratch disease, urban rabies and zoonotic ringworm) or exoanthropic zoonosis, with a sylvatic (feral and wild) cycle in natural foci outside human habitats (e.g. wildlife rabies, arbovirus, lyme disease and tularemia). Some zoonotic diseases can circulate in both urban and natural cycles (e.g. chagas disease and yellow fever).
A review study identified that 1415 species of infectious organism are pathogenic to human beings. This includes 217 viruses and prions, 538 bacteria and rickettsia, 307 fungi, 66 protozoa and 287 helminthes. Out of these, 868 (61%) are zoonotic in nature (Taylor et al. 2001). More than 60% of the emerging human infectious diseases are zoonotic in nature and 70% of their reservoirs are wild animals (Cutler et al. 2010).
The reservoirs of several zoonotic diseases are wild animals whose causative agents are viral, rickettsial, chlamydial, bacterial, parasitic and mycotic(Bengis et al. 2004). Zoonotic diseases like tuberculosis, plague and rabies have badly affected the mankind since ancient times and the reservoirs of all of these are wild animals (Stone et al. 2009). Some zoonotic diseases in human beings are self-limiting whose signs range from few days to a long term illness e.g. gastroenteritis caused byGiardia, Cryptosporidium, and Salmonella species.Some zoonotic diseases may cause abortions (Toxoplasmosis) and fatal encephalitis (Japanese encephalitis). Whereas some zoonotic diseases may causes high mortality e.g. Marburg hemorrhagic fever(MacNeil and Rollin 2012). Zoonotic diseases cause death not only in their natural hosts but also in endangered wild animal species near to extinctione.g. Ebola virus cause high mortality in monkeys (Nunn et al. 2008). It is clear from various studies in different zoos that both anthropozoonotic and zooanthroponotic transmission can occur (Adejinmi and Ayinmode 2008).
Zoonotic agents have potential to be used for bioterrorism. The bioterrorism attack is aimed to cause fear, destabilization, stress, illness and death in people, animals and plants. (Lin 2014). Air, water and food may be the warfare biological vehicles for its spread. During World War 1, anthrax was used as a biological warfare in animal populations. Glanders and typhoid were also used for bioterrorism attack in 1910 and 1970, respectively. Several cases of bioterrorism also occurred in the United States due to anthrax in September and October 2001 (Spencer 2007).
A Zoo worker should haveknowledge of the transmission of the disease to avoid its transmission. The common ways of the transmission are direct mode (ingestion, animal bites, inhalation, needle prick injuries and skin contact) and indirect mode (vector borne, fomite, long distanceand airborne transmission). In zoo management, the role of veterinarians is extremely useful. Their job exposes them to several health-related threats during routine operations. e.g. animal bites, needle prick injuries, back injuries, exposure to anesthetic gases and even mortality in certain cases (Hill et al. 1998; Kabuusu et al. 2010).
The personal protective equipment’s are not used during restraining, treatment, necropsy and cleaning the animal enclosures. It may increases the chances of zoonotic diseases to zoo workers and veterinarians. The disposal of wild animal carcasses, organs, unused food, feces and urine by unscientific methodsenhances the process of pathogens transmission(McLaughlin 2002). Laboratory personnel can also be infected with zoonotic diseases due to lack of good laboratory practices in wildlife disease diagnostic laboratories(Rietschel 1998).
Therefore, prevention and control of zoonosis must be an important part of zoo occupational health and safety measures. Preventive measures can be either general or specifically designed for a particular disease. It is possible to prevent many of the zoonotic diseases by following basic hygiene and sanitation procedures.The present study was conducted to determine knowledge, attitude, practice and experience levels about zoonosis among zoo workers of district Lahore(Lahore Zoo, Jallo Wildlife Park and Lahore Safari Zoo).
Availability: Items available for loan: UVAS Library [Call number: 2229-T] (1).
28.
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).
29.
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).
30.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Opuntia Dillenii (Ker-Gawl) Haw. Leaves Against Common Poultry Pathogens
by Sadaf Raana | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: This project was designed to evaluate the antibacterial efficacy of hexane, chloroform, ethanol and aqueous extracts of Opuntia dillenii Haw. stems against common poultry pathogens. Pathogens used were Staphylococcus aureus, Escherichia coli, Salmonella, Clostridium perfringens type A and Haemophilus species.
This study was conducted to assess antibacterial and cytotoxic activity of O. dillenii Hexane, chloroform, ethanol and water extracts were prepared and antibacterial activity was evaluated by agar well diffusion method in which zones of inhibition were measured. Minimum inhibitory concentration (MIC) of plant extracts was evaluated by micro broth dilution method. The extracts which showed the antimicrobial activity were evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell culture media was prepared and cell lines were propagated, monolayer was formed. Monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated.
O. dillenii stems extracts inhibited the growth of both Gram negative and Gram positive bacteria. Chloroform and ethanol extracts of O. dillenii showed significant antibacterial activity against all the pathogens studied as compared to hexane and aqueous extracts. Hexane extract showed maximum zone of inhibition against Haemophilus species (13mm), for chloroform extract maximum zone of inhibition was obtained for C. perfringens (25.6mm), for ethanol extract maximum zone of inhibition was obtained for C. perfringens (23.0mm) and for aqueous extract maximum zone of inhibition was obtained for C. perfringens (23.0mm). Minimum inhibitory concentration for chloroform extract was lowest for all the tested strains. For S. aureus, C. perfringens type A and S. enterica MIC was 1250μg/mL. For E. coli and
CHAPTER 6
SUMMARY
Summary
88
Haemophilus species MIC was 2083.3 and 2916μg/mL, respectively. The extracts were further investigated to test cytotoxic effect on Vero cell line using MTT assay. Only ethanol extract was observed to be cytotoxic.
Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA). The results of antibacterial activity and MTT assay were evaluated for significance of difference using analysis of variance (ANOV). The homogeneity of groups was verified by Duncan’s test at an alpha level equal to 5%.
Chloroform extract of O. dillenii stems possess antibacterial activity and can be used to design traditional medicines for the development of therapeutic agent which will be more safe, effective and economical. Availability: Items available for loan: UVAS Library [Call number: 2281-T] (1).
31.
Effect Of Oiling And Packaging On Shelf Life Of Eggs Stored At Two Different Temperatures
by Marium Munir (2008-VA-388) | Dr. Muhammad Nasir | Dr. Sanaullah Iqbal | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: In Pakistan, poultry industry is playing a vital role in the economy of the country. As eggs are perishable so it must be handled with safety. It is imperative to handle and store the eggs at appropriate conditions. But improper storage of eggs is a problem in our country which affects its quality and there is chance of microbial contamination in eggs. Oiling and packaging has variable impact on shelf life of eggs at different storage temperatures (Matt et al. 2009).Raw eggs have many benefits, they contain essential nutrients for the brain, nerves, glands and hormones, they are nutritionally balanced. raw eggs also contain an abundance of other vital substances including protein, essential fatty acids along with niacin, riboflavin, biotin, choline, vitamins A, D and E, magnesium, potassium, phosphorous, manganese, iron, iodine, copper, zinc and Sulphur. Egg yolks are one of the few foods that contain vitamin D(Watkins, 2002). As eggs are perishable food stuff,so the purposes of present research work are to analyze the effect of oiling and packaging on shelf life of eggs at two different temperatures. For this a total of 864 eggs were collected. The four different treatments were applied along with two different temperatures. Each category was further divided into four treatment strategies (108 eggs in each strategy) i.e. eggs without any treatment, oil coated eggs, eggs packed (air-tight) in white polythene bags, oil coated eggs packed (air-tight) in white polythene bags.Eggs undergone each treatment strategy were analyzed for six parameter i.e. sensory evaluation, microbial load, Physical parameters (weight, pH, egg shell percentage and haugh unit) using 18 eggs for each further divided into three replicates (6 eggs for each replicate). All the eggs were stored for 1, 7, 14, 21, 28 and 35 days.
Summary
76
Data was analyzed statistically by the 2- way ANOVA (Analysis of Variance) with 5% probability. Means was compared by DMR test.At the end of this study we were able to assess the shelf life of eggs with respect to their oiling, packaging and storage conditions. Availability: Items available for loan: UVAS Library [Call number: 2289-T] (1).
32.
Genetic Diversity Among Different Isolates Of Pasteurella Multocida From Poultry
by Arslan Sardar (2013-VA-282) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmad Anjum | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Fowl cholera is an acute bacterial disease of broiler breeders and layer breeders caused by Pasteurella multocida. In the present study, 10 isolates from different areas of Punjab were purified. These samples were confirmed by API Kit. Different molecular techniques like PCR and RFLP were used to investigate variation at the molecular level among 10 isolates collected from different areas of Punjab. Different mutations were observed among 10 field isolates at different mutation sites by sequencing. Phylogentic tree was also made using MEGA6 software that supported the sequencing results. ‘Msp1’ endonuclease cleaved bacterial whole genome at different cutting sites, all 10 isolates collected from different districts of Punjab cleaved into 3 to 5 fragments ranging from 600 to 10000 base pairs which showed the genetic variation among 10 isolates of P.mulocida. Availability: Items available for loan: UVAS Library [Call number: 2315-T] (1).
33.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Zingiber Officinale Rhizome Against Common Poultry Pathogens
by Ghalia Qayyum (2013-VA-779) | Dr. Aqeel Javeed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Antimicrobial compounds having plant origin inhibit bacteria through different mechanisms and can be used for the treatment of infections against resistant microbes. Majority of antibacterial drugs in clinical use are derived from natural origin. Hence, the present study is designed for antibacterial and cytotoxic evaluation of different extracts of Zingiber officinale rhizome against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale were prepared by soxhlet extraction. Antibacterial activity of these extracts was determined by agar well diffusion method against Staphylococcus aureus, Clostridium perfringens type A, Escherichia coli, Salmonella enterica and Haemophilus paragallinarum. Zone of inhibitions were determined by well diffusion method. MICs of plant extracts were determined by micro broth dilution method. Cytotoxic activity was evaluated by applying MTT assay on Vero cell lines
The zone of inhibitions showed by hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale against Staphylococcus. aureus were 12.33mm, 13.67mm, 16.33mm and 14mm; against Clostridium perfringens type A were 12mm, 16.33mm, 14mm and 8.33mm; against Escherichia coli were 14.33mm, 13.33mm, 14.33mm and 12mm; against Salmonella enterica were 17mm, 17.33mm and 12mm; against Haemophillus paragallinarum were 12.67mm, 13mm and 14mm respectively. Hexane extract showed no zone of inhibition against Salmonella enterica and aqueous extract was ineffective against Haemophillus paragallinarum.
MICs values of hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale rhizome against Staphylococcus. aureus were 2500µg/ml, 625µg/ml, 2500µg/ml and 2083.33µg/ml; against Clostridium perfringens type A were 2500µg/ml, 312.5µg/ml, 1250µg/ml and 5000µg/ml; against Escherichia coli were 5000µg/ml, 1250µg/ml, 5000µg/ml and 5000µg/ml; against Salmonella enterica were 312.5µg/ml, 5000µg/ml, 5000µg/ml; against Haemophillus paragallinarum were 2500µg/ml, 1458.33µg/ml and 2500µg/ml respectively. MIC was not performed against hexane extract of Salmonella enterica and aqueous extract of Haemophillus paragallinarum as no zone of inhibition observed against them. Hexane extract of Zingiber officinale rhizome was cytotoxic at concentration ≥ 750µg/ml, chloroform extract at concentration ≥ 1500µg/ml and aqueous extract at concentration ≥5000µg/ml. Ethanol extract at concentration ranging from 1500µg/ml to 2.92µg/ml was not cytotoxic to cell.
The indigenous plant Zingiber officinale have antibacterial activity against common poultry pathogens and helpful to develop new drug from plant origin.
Availability: Items available for loan: UVAS Library [Call number: 2324-T] (2).
34.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Zingiber Officinale Rhizome Against Common Poultry Pathogens
by Shumaila Nawaz (2013-VA-442) | Dr. Muhammad Adil Rasheed | Dr. Aqeel Javeed | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Plants produce a diverse range of bioactive molecules, making them rich source of
different types of medicines. Calotropis procera, a giant milk weed, is known for its
pharmacological importance for centuries. This shrub has been known to possess analgesic,
antitumor,
antihelmintic,
antioxidant,
hepatoprotective,
anti-diarrhoeal,
anticonvulsant,
antimicrobial, oestrogenic, anti-nociceptive and anti-malarial activity. A very little information is
available regarding the antibacterial and cytotoxic activity of Calotropis procera so the present
study is designed to evaluate the antibacterial and cytotoxic activity of this plant. This study was
conducted to access antibacterial and cytotoxic activity of Calotropis procera.
Hexane, chloroform and ethanol, aqueous extracts were prepared by sequential extraction
method and antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli,
Salmonella enterica, Clostridium perfringens type A and Haemophilus paragallinarum by agar
well diffusion method in which inhibitory zones were measured. The extracts which showed the
antimicrobial activity were evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell
culture media was prepared and cell lines were propagated, monolayer formed. Monolayer was
exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival
percentage was calculated.
Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for
windows version 16, SPSS inc, Chicago, IL, USA). Results will be compared using one way
ANOVA analysis.
102
SUMMARY
Chloroform and ethanol extracts of Calotropis procera leaves have antibacterial activity.
It may help to design traditional medicines for the development of therapeutic agent which will
be more safe, effective and economical.
Availability: Items available for loan: UVAS Library [Call number: 2330-T] (1).
35.
Status Of Brucellosis And Its Effect On Hemogram And Serum Biochemistry In Indigenous, Cross-Bred And Exotic Dairy Cattle Herds
by Muhammad Hareem Afzal (2008-VA-250) | Dr. Muhammad Avais | Dr. Jawaria Ali Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Brucellosis mainly infects food animals such as cattle, buffalo, goats and sheep. Brucella abortus is the principal cause of brucellosis in cattle and is shed from the infected animal at or around the time of calving or abortion. The present study was conducted on 450 animals on three different strains/breeds of cattle i.e. Exotic (150), Cross-bred (150) and local cattle (150) from 10 different privately owned livestock farms of varying holdings of district Lahore. An epidemiological questionnaire focusing on herd traits as well as husbandry and sanitary practices that could be associated with the risk of Brucellosis infection was completed. Serum samples were collected and analyzed using Rose Bengal Plate Test (RBPT). The serum samples positive for Brucellosis through RBPT further subjected to Serum Agglutination Test (SAT). To check the effect of Brucellosis on hemogram, blood samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis were collected and processed for TLC, DLC, RBC, Hb, MCV, MCHC MCH and platelets using automated haematology analysed at UDL, UVAS, Lahore. Similarly, to see the effect of Brucellosis on Serum biochemistry, serum samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis collected and analysed for glucose, total protein, albumin, Creatinine, Alanine Aminotransferase (ALT), Aspartate Aminotranferase (AST) and Sorbitol Dehydrogenase (SD) using commercially available kits.
Summary
62
RBPT revealed overall prevalence 17.7% higher than SAT 10.6%. Prevalence of brucellosis is higher in Cross-Bred (22.7%) followed by local cattle (18.9%) and exotic (12%).
Hemato-boichemical results showed that increase in TLC, MCV While slight changes in Hb, MCHC, RBC and values of MCV stays within normal range. On the other hand serum biochemistry increase in AST while decrease in ALT and SD found. Availability: Items available for loan: UVAS Library [Call number: 2348-T] (1).
36.
Mutational Screening Of The RB1 Gene In Pakistani Patients With Retinoblastoma
by Saeeda Kalsoom (2007-VA-555) | Dr. Muhammad Wasim) | Dr. Khushnooda Ramzan | Dr. Ali Raza Awan | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Retinoblastoma is a neonatal intraocular tumor caused by biallelic inactivation of RB1 gene. Rb
patients and asymptomatic carriers undergo a series of clinical tests for diagnosis and tumor
treatment. These clinical examinations prove to be expensive and time consuming. On the other
hand if the proband’s RB1 gene mutation status is determined by genetic testing, it can prove as
more significant and cost effective diagnostic methods. Secondly, only those asymptomatic or at
risk carriers with the mutation, require clinical surveillance while those proven to be unaffected
do not require additional clinical examinations. Furthermore early diagnosis of Rb by molecular
testing can enable and enhance clinical management, earlier treatment, follow-up care, carrier
screening, genetic counseling, prenatal diagnosis and reproductive planning in predisposed
families. Irrespective of the importance of molecular testing of Rb patients, in Pakistan only a
few clinical reports on Rb are available so, there was a dire need to find RB1 mutations in
Pakistani Rb patients and to set a molecular based diagnosis for poor affected families. Keeping
in view the importance of molecular diagnosis, in this study a reliable genetic test has been
developed to detect the RB1 germline mutations in Pakistani Rb patients.
During this study, 70 Rb patients including 38 unilateral and 32 bilateral cases were enrolled,
from different regions of Pakistan. By using direct sequencing method, seven novel and twelve
reported RBI mutations were found. The novel mutations included three frameshift mutations
(c.1116_1119delCACT in exon 11, c.1436_1437delAC in exon 16 and c.2060_2061insTCATT
in exon 20) and four substitutions (c.148G>T in exon 2, c.610G>T in exon 2, g.94G>C in exon
7, c.947A>T in exon 10 and g.1991G>C in promoter region) while twelve reported mutations in
146
22 patients included, 9 substitutions (c.160G>T in exon 2, c.289G>T in exon 3, c.751C>T in
exon 8, c.920C>T in exon 9, c.967G>T in exon 10, c.1072C>T in exon 11, c.1654C>T in exon
17, c.2063T>C in exon 20 and c.2359C>T in exon 23), one frameshift mutation (c.772_776del in
exon 8) and two splice site mutations (c.380+1G>T and c.1215+1G>A in intron 3 and 12
respectively). Mutation detection rate was found to be 77.8% in (7/9) bilateral familial, 50% in
(2/4) unilateral familial, 56.5% in (13/23) bilateral sporadic and 14.7% in (5/34) unilateral
sporadic patients while overall rate of mutations in bilateral and unilateral patients was detected
as 62.5% (20/32) and 18.4% (7/38) respectively. Beside mutations one novel c.940-64C>T
(intron 9) and nine reported intronic variants c.380+45 C>T (intron 3), c.501-77G>A (intron 4),
c.1128-72T>G (intron 11), c.1695+99A>T (intron 17), c.1695-1696delAA (intron 17), c.1815-
104A>G (intron 18), c.1961-10T>C (intron 19), c.2663+33T>C (intron 25) and c.2664-10T>A
(intron 25) were also found. Carrier screening facility was also provided to six asymptomatic
siblings (as possible carriers) of familial proband but none of them was found to be diseased.
Hopefully, in future the findings and developed protocol of this study will help to reveal the
molecular basis of Rb in Pakistani Rb patients which additionally help to secure vision and life
of Rb patients. Further, in Pakistan there is dire need to develop “National Rb Registry Centre”,
to register all new Rb cases for finding incidence rate and prevalence of Rb in Pakistan. Beside
this other related issues like financial constraints, health education, planning and awareness
about Rb, occupational training for health providers, capacity building for neonatal
ophthalmologic screening and cosmetic rehabilitation for surviving Rb patients are important and
should consider. Availability: Items available for loan: UVAS Library [Call number: 2370-T] (1).
37.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Eucalyptus Globulus Leaves Against Common Poultry Pathogens
by Asma Iqbal (2013-VA-563) | Dr. Muhammad Adil Rasheed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Phytomedicines mark the major component of health care as natural medicines have always provided the strong foothold for the discovery and manufacturing of synthetic drugs. So plants are a rich source of bioactive compounds having many therapeutic activities and majority of them are still untapped. Eucalyptus globulus is a medicinal plant known for its value to cure asthma, respiratory infections, cough and allergic reactions. The antimicrobial activity, insecticidal and hypoglycemic activity have also been credited to the plant. Most of the studies have been conducted on the essential oils of Eucalyptus globulus and little work has been reported on extracts. Whereas, sequential extracts has not been employed yet.
Hexane, chloroform and ethanol, aqueous extracts were prepared by the sequential extraction on Soxhlet apparatus and antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli, Salmonella enterica, Clostridium perfringens type A and Haemophilus paragallinarum by agar well diffusion and micro broth dilution method. The zones of inhibition and minimum inhibitory concentration were determined. The extracts showing antimicrobial activity were further evaluated for cytotoxicity by using MTT assay on Vero cell line. The cell culture media was prepared and cell lines were propagated to form monolayer then monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated.
The statistical analysis was conducted with help of Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA) and results were compared using one way ANOVA.
Summary
89
The zones of inhibitions showed by hexane, chloroform, ethanol and aqueous leaf extracts of Eucalyptus globulus against Staphylococcus aureus were 0.0, 19.3, 20.3 and 23.3mm; against Clostridium perfringens type A were 14, 22.3, 14.0 and 15.3mm; against Escherichia coli were 0.0, 12.6, 13.3 and 15.6mm; against Salmonella enterica were 10, 12.3, 18.6 and 21mm; against Haemophilus paragallinarum were 0.0, 8.6, 14 and 18mm respectively. Hexane extract showed no zone of inhibition against Staphylococcus aureus, Escherichia coli and Haemophilus paragallinarum.
The MICs values of hexane, chloroform, ethanol and aqueous leaf extracts of Eucalyptus globulus against Staphylococcus aureus were 0.00, 104.1, 32.55 and 312.5 μg/ml; against Clostridium perfringens type A were 52.08, 39.06, 16.27 and 312.5 μg/ml; against Escherichia coli were 0.00, 78.12, 260.4 and 625.0 μg/ml; against Salmonella enterica were 13.02, 104.1, 130.2 and 416.6 μg/ml; against Haemophillus paragallinarum were 0.00, 104.1, 260.4 and 416.6 μg/ml respectively. MIC was not performed against Staphylococcus aureus, Escherichia coli and Haemophilus paragallinarum for hexane extract as no zone of inhibition was observed against them.
Hexane extract of Eucalyptus globulus was cytotoxic at concentration ≥ 312.5μg/ml, chloroform extract at concentration ≥ 375μg/ml, ethanol extract at concentration ≥ 625μg/ml and aqueous extract was cytotoxic at concentration ≥312.5 μg/ml.
The indigenous plant Eucalyptus globulus has antibacterial activity against common poultry pathogens and can be helpful for development of new drugs of plant origin. Availability: Items available for loan: UVAS Library [Call number: 2429-T] (1).
38.
Effect Of Bio-Stimulation On Estrus Expression And Pregnancy Rate In Cidr Based Synchronization Protocol In Nili-Ravi Buffalo
by Abdul Waheed (2009-VA-133) | Dr. Aijaz ali Channa | Dr. Syed Murtaza Hassan Andrabi | Prof. Dr. Mian Abdul Sattar | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Our water buffalo (Bubalus bubalis) has much potential for production of milk. But this
animal has some problems regarding reproduction including delayed puberty, poor estrus behavior,
silent heat, long postpartum period and low conception rate by artificial insemination. This leads
to poor reproduction and hence great economic loss. Therefore, the requirement is to address these
problems efficiently and formulate more effective techniques for improvements. Researchers have
devised many estrus synchronization protocols (PGF2α, P4, GnRH, eCG, hCG etc.) that help
bringing many animals in heat and hence improve the reproductive performance when fixed time
artificial insemination is combined with them. But these protocols give inconsistent results when
they are applied on buffaloes making it necessary to improve the techniques.
This study was planned on the hypothesis that presence of bull (bio-stimulation), at the
time of synchronization, may play an important role in enhancement of estrus intensity and fertility
rate in Nili-Ravi buffaloes. Seventy one adult buffaloes were randomly selected from different
areas of field conditions and LRS (NARC) and subjected to CIDR based heat synchronization in
combination of either bio-stimulation or non-stimulation. The animals were observed for
behavioral estrus signs twice a day starting after 12 hours of CIDR removal till 96 hours.
Pregnancy diagnosis was done by rectal palpation 60 days post CIDR removal. Estrus response
and pregnancy rate were analyzed by Chi-square test using MINITAB version 15. Estrus signs and
total estrus intensity were compared by Mann Whitney U test. Difference was considered
significant at probability level of (P < 0.05).
In peri-urban areas, more animals from bio-stimulated group showed better behavioral
estrus signs, more total intensity score and significantly higher pregnancy rate as compared to nonSUMMARY
63
stimulated group of animals. At LRS (NARC), more animals from non-stimulated group were
found in behavioral estrus but intensity of heat signs was high in bio-stimulated animals.
Pregnancy rate was also higher in non-stimulated animals but the difference was not significant.
Overall, in this study, we got higher pregnancy rate in bio-stimulated animals than non-stimulated
group which indicates a positive response of bull stimulation on reproductive performance of Nili-
Ravi buffaloes who were synchronized with CIDR based estrus synchronization protocol. Availability: Items available for loan: UVAS Library [Call number: 2469-T] (1).
39.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Astragalus Membranaceus Roots
by Sadia Alvi (2013-VA-595) | Dr. Aqeel Javeed | Dr. Muhammad Ovais Omer | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The present study was designed to evaluate antibacterial and cytotoxic evaluation of different extracts of Astragalus membranaceus root against common poultry pathogens. Sequential extraction with hexane, ethanol, chloroform and aqueous solvents was prepared and antibacterial activity was evaluated by using agar well diffusion. Minimum inhibitory concentration (MIC) of plant extracts was evaluated by micro broth dilution test. The extracts exhibiting antimicrobial activity were further evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell culture media was prepared and cell lines were propagated, monolayer was formed. This monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated. Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA). Results of antibacterial activity and MTT assay were compared using DMR posthoc test. Growth of Clostridium perfringens, Escherichia coli, Haemophilus species, Salmonella enterica and Staphylococcus aureus inhibited by all extracts of Astragalus except aqueous extract which shows no zones of inhibition against C. perfringes. MIC values were higher for aqueous extract against all selected bacteria and lowest for chloroform against E. coli, S. enterica and Staph. aureus (208.3ug/ml, 156.25ug/ml, 78.125ug/ml respectively) for hexane against Haemophilus species (833.3ug/ml) and for all three extracts against C.perfringes (1250ug/ml). Hexane, chloroform and ethanol extracts were appeared to be safe at all concentrations except ≥ 2000μg/ml, ≥1000μg/ml and ≥3000μg/ml respectively while aqueous extracts showed cytotoxicity at concentrations ≥625μg/ml. Astragalus membranaceus
SUMMARY
104
showed antibacterial activity against all selected pathogens. Chloroform and hexane extracts showed greater antibacterial activity than ethanol and aqueous. Cytotoxicity values for chloroform extract are safer than rest of three extracts. Astragalus membranaceus may be used to design traditional medicines for the development of therapeutic agent which will be more safe, effective and economical. Availability: Items available for loan: UVAS Library [Call number: 2444-T] (1).
40.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Ocimum Basilicum Leaves Against Common Poultry Pathogens
by Shomaila Naz (2013-VA-1001) | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Antimicrobial compounds having plant origin inhibit bacteria through different mechanisms and can be used for the treatment of infections against resistant microbes. Majority of antibacterial drugs in clinical use are derived from natural origin. Hence, the present study is designed for antibacterial and cytotoxic evaluation of different extracts of Ocimum basilicum seeds against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Ocimum basilicum leaves and seeds were prepared by soxhlet extraction. Antibacterial activity of these extracts was determined by agar well diffusion method against Staphylococcus aureus, Clostridium perfringens type A, Escherichia coli, Salmonella enterica and Haemophilus paragallinarum. Zone of inhibitions were determined by well diffusion method. MICs of plant extracts were determined by micro broth dilution method. Cytotoxic activity was evaluated by applying MTT assay on Vero cell lines. All the results were statistically analyzed by one way ANOVA and compared means by Duncan’s multiple range of posthoc test at significance level of P≤0.05.
The results of zone of inhibitions showed by Ocimum basilicum leaves and seeds extracts ranging from 11.33-20.0 mm values of MIC results ranging from 4.889 μg/ml-2500 μg/ml of hexane, chloroform and ethanol. The aqueous extract of Ocimum basilicum have no activity against any bacterial pathogen. Ethanol extract of Ocimum basilicum leaves was cytotoxic at 500 μg/ml. Hexane extract of Ocimum basilicum seeds was cytotoxic at concentration ≥625 μg/ml, chloroform at concentration ≥19.53 μg/ml and ethanol extract at concentration ≥750 μg/ml.
The indigenous plant Ocimum basilicum have antibacterial activity against common poultry pathogens and helpful to develop new drug from plant origin. Availability: Items available for loan: UVAS Library [Call number: 2443-T] (1).
41.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Glycyrrhiza Glabra (Liquorice) Roots Against Common Poultry Pathogens
by Javaria Arooj (2013-VA-596) | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Majority of antibacterial drugs in clinical use are derived from natural origin. Hence, the
present study is designed for antibacterial and cytotoxic evaluation of different extracts of
Glycyrrhiza glabra Linn. roots against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Glycyrrhiza
glabra Linn. roots were prepared by soxhlet extraction. Antibacterial activity of these extracts was
determined by agar well diffusion method against Staphylococcus aureus, Clostridium perfringens
type A, Escherichia coli, Salmonella enterica and Haemophilus paragallinarum. Zone of
inhibitions were determined by well diffusion method. MICs of plant extracts were determined by
micro broth dilution method. Cytotoxic activity was evaluated by applying MTT assay on Vero
cell lines.
The zone of inhibitions showed by hexane, chloroform and ethanolic extracts of
Glycyrrhiza glabra Linn. roots against Staphylococcus. aureus were10.3mm, 13.0mm, 11.6mm;
against Clostridium perfringens type A were20.0mm, 17.3mm, 17.3mm; against Escherichia coli
were11.6mm, 19.3mm, 16.0mm; against Salmonella enterica were13.6mm, 14.0mm,14.0mm;
against Haemophillus paragallinarum were13.0mm, 15.0mm, 17.0mm respectively. Aqueous
extract showed no zone of inhibition against any test bacteria.
MICs values of hexane, chloroform and ethanolic extracts of Glycyrrhiza glabra Linn.
roots against Staphylococcu aureus were 13.0μg/ml, 312.5μg/ml and 104.1μg/ml; against
Clostridium perfringens type A were 9.766μg/ml, 71.61μg/ml and 520.8μg/ml; against
Escherichia coli were 65.1μg/ml, 52.8μg/ml and 156.25μg/ml; against Salmonella enterica were
Summary
86
19.5μg/ml, 130.2μg/ml and 78.12μg/ml; against Haemophillus paragallinarum were 91.1μg/ml,
29.2μg/ml and 130.2μg/ml respectively. Aqueous extract showed no MIC value as no zone of
inhibitions wereobserved against them. Hexane extract of Glycyrrhiza glabra Linn. roots was
cytotoxic at concentration ≥ 650μg/ml, chloroform extract at concentration ≥ 2500μg/ml and
ethanolic extract was not cytotoxic to cell.
The indigenous plant Glycyrrhiza glabra Linn. roots have antibacterial activity against
common poultry pathogens and helpful to develop new drug from plant origin. Availability: Items available for loan: UVAS Library [Call number: 2442-T] (1).
42.
Characterization And Thermostability Of Phytase Produced By Indigenous Aspergillus Niger Isolates
by Madeeha Tariq (2010-VA-293) | Prof. Dr. Aftab Ahmad Anjum | Dr. Jawad Nazir | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Phytase enzyme now becomes more important commercially. Presence of phytate in food
and feed make them less nutritive due to phytate complexes mainly with mineral ions and
proteins. Phytase in monogastric animals and human stomach either produced in small amount or
not. This leads to phosphorous Pi deficiency. Supplementation of food and feed with phytase
enzyme full fill this deficiency through degradation of phytate complexes and release of Pi.
Degradation of phytate complexes makes phosphorous other mineral ions and amino acids
available for growth and development. It was proved that feed conversion rate in poultry
increased due to supplementation of phytase in poultry feed. Feed of monogastric animals mostly
at industrial level pelleted to give it a shape or to kill microorganisms (sterility). At industrial
level enzyme production and processing cost about 2 billion. So this demands a thermostable
phytase to use at industrial level or its cost effective production.
Aspergillus niger have been used industrially for production of beneficial enzymes. A.
niger isolates procured from department of microbiology were confirmed through macro and
microscopic characteristics as A. niger. These isolate were screened for phytase production on
phytase screening medium PSM agar. Positive isolates identified through noval staining using
2% cobalt chloride, 6.25% ammonium molybdate and 0.42% ammonium vanadate for contrast.
Positive isolates next proceeded for phytase enzyme production in broth media (pH 5.6) using
0.5% sodium phytate as substrate. Incubation was done at 30oC for 5-7 days in shaking incubator
150rpm. After production quantification of enzyme was carried out through enzyme activity
assay. There maximum (274.99±10.14 FTU/ml) and minimum (68.88±2.55 FTU/mL) activity of
phytases from isolate PASN01 and PASN08 was observed. Phytases characterized through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) to know protein
molecular weights. Highest molecular weight 107.82kDa was PASN06 and lowest was 35.21kDa
of PASN 01.
Aspergillus niger spores subjected to steam heat treatment at 30oC, 45oC, 60oC, 75oC and
90oC for 15, 30, 45, 60minutes to identify thermostability. At 30oC and 45oC temperature, spores
of A. niger isolates found to be thermostable. But at 60oC, 75oC, or 90oC treatment spores
become inactivated or there 6.0 logarithmic reduction in spore count was observed.
Thermostability of phytases was found at 60oC, 75oC, 90oC for 15, 30, 45, and 60
minutes treatments. Enzyme from A. niger PASN01 and PASN08 observed as thermostable at
60, 75 and 90oC. Phytases from PASN01 and PASN08 showed 160.55±42.96 and 00±.00
FTU/mL decreased in activity after 45 minutes of treatment at 60oC temperature, respectively.
PASN01 phytase displayed 163.88±23.35, 172.77±7.52 and 171.66±7.26 FTU/mL decreased in
activity after 60minutes treatment at 60, 75 and 90oC. In case of PASN08 phytase at 60, 75 and
90oC temperature after 60minutes treatment, 13.33±10.41, 16.66±6.00 and 23.88±41.37 FTU/mL
decreased in activities were observed, respectively. PASN08 phytase observed more
thermostable than other phytases of A. niger isolates. Enzyme can bear pelleting and pre
pelleting temperatures. Enzyme from PASN08 also observed stable during storage at room
temperature.
Conclusion:
A. niger PASN08 spores inactivated or killed and phytase observed stable at 60oC
temperature, after 60mins treatment. Temperature 60oC may be used industrially for cost
effective thermostable phytase production from indigenous A. niger isolate PASN08. Availability: Items available for loan: UVAS Library [Call number: 2475-T] (1).
43.
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: ; 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).
44.
Isolation And Antibiotic Resistance Profiling Of Enterococcus Faecium Recovered From Retail Fish In Lahore City
by Maria Butt (2010-Va-281) | Dr. Ali Ahmad Sheikh | Prof. Dr. Aftab Ahmad Anjum | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Enterococcus faecium is an enteric, gram positive and lactic acid bacteria which belongs to genus enterococcus and inhabit the intestinal tract of human, fish and other warm blooded animals. Due to irrational use of antibiotics in human and veterinary sector, antibiotic resistance has been developed in commensal bacteria including Enterococcal species. These resistant bacteria are released in environment through human and animal waste and transfer resistant genes to susceptible bacteria present in wetlands making them antibiotic resistant. E. faecium is considered to be involved in transmission of resistance genes, present on mobile genetic elements through conjugation to other bacteria. The resistant bacteria can be transferred to human through food chain. The present study was designed to evaluate the prevalence of E. faecium recovered from retail fish samples collected from various areas of Lahore city. Antibiotic resistance profiling of the isolates against commonly used antibiotics was also determined.
In current study 65 fish samples (intestinal swabs) were processed for isolation of E. faecium through standard culturing and biochemical reactions. Out of 65 swab samples, 30 samples (47.69%) were found positive for Enterococcus faecium. Antibiotics resistance profiling showed that the isolates were resistant to antibiotics mentioned as below: Ampicillin (100%) > erythromycin (56.6%) > rifampicin (53.3%) > Chloramphenicol (30%), ciprofloxacin (30%) > tetracycline (20%), vancomycin (20%) > Teicoplanin (13.3%) > Doxycyclin (6.6%) > Fosfomycin (0%). E. faecium isolates showed resistant to at least 2 or 3 antibiotics of different group. In conclusion it is observed that retail fish is the carrier of antibiotic resistant Enterococcus faecium and
Summary
51
could transfer resistant genes to wetlands and other aquaculture from where it could be transferred to human body. Efforts should be made to use antibiotics wisely and hygienic practices should be followed during slaughtering and processing of fish meat to avoid bacterial spread from animal source to human beings. Availability: Items available for loan: UVAS Library [Call number: 2493-T] (1).
45.
Evaluation Of Antibacterial Effect Of Gymnema Sylvestre Species Cultivated In Pakistan
by Muhammad Tahir (2011-VA-339) | Dr. Muhammad Adil Rasheed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This study was conducted to determine the use of medicinal plants as an antibacterial agent and its potential to as an alternative medicine against bacterial infections. For this purpose Sequential extracts (i.e. Hexane, Chloroform, Ethanol and Aqueous) of Gymnema sylvestre R.Br. were tested against S. aureus, E. coli, S. enterica, C. perfringens type-A, H. paragallinarum. Of each bacterium 3 isolates were tested by using well diffusion method. The results were obtained by determining the ZOI by well diffusion method and MIC by using 96 well ELISA plate.
The mean ZOI and mean MIC values of G. sylvestre leaves extracts showed that chloroform and ethanolic extracts have more antibacterial activity against all five microorganisms. Only chloroform and ethanolic extracts showed antibacterial activity against all 5 microorganisms while hexane extract showed antibacterial activity against S. enterica, S. aureus, H. paragallinarum, C. perfringens type- A but no activity was observed against E. coli. On the other hand aqueous extract have showed antibacterial activity only against C. perfringens type-A but no antibacterial activity against remaining four bacteria under study. While analyzing results based upon MIC, the chloroform extract has more antibacterial effect when compared with hexane. Hexane extract was more potent than aqueous extract whereas ethanolic extract was the least potent.
When overall antibacterial effect of all the extracts was evaluated against all bacterial strains, it was observed that C. perfringens type-A was the bacterium most vulnerable to antibacterial activity of sequential extracts of dried leaves of G. sylvestre as it responded
Summary
94
to all four sequential extracts and gave maximum zone of inhibition (10-22mm range) while no other bacteria showed such bigger zone of inhibition.
On the basis of MIC, it can be assumed that chloroform extracts have more antibacterial components as compared to hexane extract. Hexane extracts have more antibacterial components as compared to ethanolic extracts. The activity of aqueous extracts is negligible as it showed response against only one bacterium.
MTT assay was performed on supersaturated solutions of sequential extracts of dried leaves of G. sylvestre. Results revealed that small concentrations of these extracts are not toxic. Cell survival percentage (CSP) values below 50% were given at concentrations of 5800μg/ml (38.76%), 7225μg/ml (43.71%), 8150μg/ml (44.90%) and 3125μg/ml (41.84%) by hexane, chloroform, ethanolic and aqueous extracts respectively. Finally, on the basis of MIC and CSP for all of four sequential extracts, it is concluded that chloroformic extract is the most active and safe extract against all of 5 experimental bacteria, while hexane extract is safe against only C. perfringens type-A and ethanolic and aqueous extracts are cytotoxic on their MIC values for all the experimental bacteria. Statistical analysis showed that ZOI and MIC values were significantly different between the groups while within the same group they were non-significant.
Finally it can be concluded that the leaves of plant Gymnema sylvestre R.Br. cultivated in Pakistan has considerable antibacterial activity and considerable safety profile so it must be further studied, characterized, purified and chemically isolated so that may be converted to proper dosage form and this miracle plant may be used therapeutically to cure various ailments including bacterial infections especially poultry infections. Availability: Items available for loan: UVAS Library [Call number: 2503-T] (1).
46.
Assessment Of Afflatoxins Contamination In Peanuts
by Zanib Hashmi (2009-VA-512) | Dr. Naureen Naeem | Dr. Sanaullah Iqbal | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Peanut is the most important agricultural crop of Pakistan. Peanut is a dicotyledonous, herbaceous, pubescent, rigid or low growing plant and the only species cultivated is (Arachishypogaea L.). Peanut is rich in protein, fat and carbohydrates, some percentage of Ca, K, P, Mg and vitamin E is also present. Peanut is an excellent source of edible oil as it contains about 50 to 53 percent good quality oil used in ghee, margarine and salad. There is high risk of contamination of peanuts with aflatoxins(AFB1, AFB2, AFG1 and AFG2) because of fungal attack during the drying of peanut pods. Out of all these aflatoxins AFB1 is most important. Aflatoxins are toxic, carcinogenic secondary metabolites of Aspergillusflavus, Aspergillusparaciticus and Aspergillusnomius. Aflatoxins can cause illness to human results in Aflatoxicosis. Aflatoxins are carcinogenic compounds that are causative agents in human hepatic and extra hepatic carcinogenesis. The chief attacking organ for aflatoxins B1 toxicity and carcinogenicity is liver. From the safety point of view aflatoxin management is important for the production of safe and excellent quality peanuts.
For this purpose present study was conducted to determine the level of aflatoxins in peanuts (roasted, un-roasted). Samples will be collected/purchased by simple random collection technique from local markets and vendors from different areas ( Sabzazar, Wahdat road , Shad bagh, Data darbar, Akbarimandi, Beaden road, Lohari gate, Ek-moria pull, Liberty, Firdous market, Siddiqiacoloney, Mughal pura, Faizbagh, Rehmanpura, Gulberg, Model town, Islam pura, Shahdara, Rang mahal, Muslim town, Township, Iqbal town, Awan town, Niazbegh, Mozang, Outfall road, Sanatnagar, Cantt, Secretriate and Shad man) of Lahore. The samples were analyzed by thin layer chromatography (TLC) to check the presence of aflatoxins (B1, B2,
G1 and G2). TLC analyses were further confirmed by high performance liquid chromatography (HPLC) to verify the accuracy of TLC. These analyses were performed in the Department of Food Science and Human Nutrition and WTO labs, University of Veterinary and Animal Sciences, Lahore. As out of 120 total samples of peanuts 60 samples were taken from vendors with 2 categories of roasted and unroasted while 60 samples were collected from shops with the same categories. Out of 120 samples, 55 (45.8%) were contaminated. In these 55 samples 48 (87.2%) samples were contaminated with aflatoxin B1.Aflatoxin G1 is also present in 3 samples (5.45%), aflatoxin B2 in 3 (5.45%) samples and Aflatoxin G2 is present only in one samples collected from vendors, and we can say that 1.8% samples were contaminated with aflatoxin G2.
Present study will be supportive for the investigation of aflatoxins in peanuts. Peanuts are widely consumed all over the world and occurrence of aflatoxins in this commodity is a major concern to human health. The present situation is too much worse about the levels of aflatoxins which are higher than the prescribed limit by the regulatory authorities. It was observed that TLC technique is good for the determination of aflatoxins in developing countries where the facilities of sensitive instruments are not accessible. Furthermore to quantify levels of aflatoxins by using sensitive instruments like HPLC, GC-MS and LC-MS is required for accurate detection of Aflatoxins in peanuts in markets to protect the consumers from exposure of aflatoxins high level which are carcinogenic and hepatotoxic.
Availability: Items available for loan: UVAS Library [Call number: 2614-T] (1).
47.
Effect Of Garlic And Ginger Extract On The Shelf Life Of Fish
by Dure-e-Shahwar (2009-VA-439) | Dr. Naureen Naeem | Dr.Sanaullah Iqbal | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The fish is highly perishable food which contains high protein and omega3 fatty acids. It contain enzyme which cause autocatalysis of muscles after harvesting. Due to lack of Knowledge and poor storage and handling practices cause fish spoilage and deterioration of fish.
Ginger and garlic are spices, also contain a variety of bioactive substances which are of considerable use from the standpoint of food science and technology. Ginger and garlic shows excellent inhibition against food pathogens such as Staphylococcus aureus; Bacillus spp., Escherichia coli and Salmonella spp.
Antimicrobial properties of garlic and ginger may control the microbial growth of fish and is able to minimize fish spoilage. Fish was taken from fish farm then washed and cleaned, cut the fish and left at room temperature for water dropping then weighed it. Each sample was containing 20gm weight.
Then dipped samples in extract of ginger and garlic that have doses 15%, 20 %, 25%for ninety minute, then was wrapped in polythene bag and put in refrigerator for 5 months.
Aerobic plate count was performed after fortnightly by the method of standard plate count and assessed sensory condition of fish by sensory evaluation after one month.
In control group, the Bacillus cereus significantly increased with time (during storage) While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life and Bacillus cereus significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. Garlic was more effective then ginger in separately treatment.
In control group, staphylococcus significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life of staphylococcus significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. In comparison to garlic, ginger was observed most efficient in controlling staphylococcus growth in fish samples.
In control group, Salmonella significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life and Salmonella significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. Seprately20 % garlic and ginger show same result.
In control group, Streptococcus significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life and Streptococcus significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group. But garlic showed better results as compared to the ginger in respective concentrations.
In control group, Shigella significantly increased with time (during storage). While, in all other treatments both garlic and ginger treatment prove effective to increase shelf life Shigella significantly decreased with time (during storage). The apparently huge decreased was observed in combined form of ‘25% Garlic & Ginger’ treatment group.
The sensory evaluation results showed that with increasing concentration of ginger and garlic separate and in combination of both have profound effects on sensory parameters. It is evident
Summary
63
from the results after five months of trial that garlic and ginger can be used to control microbial growth in fish samples and their acceptability on sensory scale is better than the control samples. Treated samples were more liked and observed acceptable according to grading scale. By comparing the whole results of sensory evaluation it has become very easy to access the positive outcomes of the applications of ginger and garlic in different concentrations and in combination. Ginger and garlic in combination were more liked and maintained their color, juiciness, flavor, tenderness and oiliness level.
Data was statistically analyzed by applying 2 Way ANOVA. There was mean score difference (p<0.05) among garlic treatment, ginger treatment and combination of garlic and ginger treatment with bacterial count. But ginger has least effect as compare to garlic but in combination they became more effective against bacterial count. There was mean score significant difference (p<0.05) among treatment and time with sensory evaluation.
This study shows that combination of both spices 25% ginger & garlic is more effective then separately ginger & garlic. Garlic shows better result against control of bacterial count Streptococcus and Bacillus cercus. Ginger shows better result against control of bacterial count in Staphylococcus and Shigella. Both spices show almost same control of bacterial count against Salmonella. Availability: Items available for loan: UVAS Library [Call number: 2611-T] (1).
48.
Chemical, Microbiological And Toxicological Evaluation Of Textile Dyeing Industry Wastewater
by Muhammad Furqan Akhtar (2011-VA-265) | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Exposure to complex mixtures like textile effluent poses risks to animal and human health such as mutations, genotoxicity, pathological lesions and oxidative damage. The aim of the present study was to quantify metals and identify organic pollutants in untreated textile dyeing industry wastewater, to determine the bacterial load of wastewater, isolate and identify heavy metals tolerant bacteria and to determine its mutagenic, genotoxic and cytotoxic potential, influence on normal physiology and effects on oxidative stress biomarkers in effluent exposed rats.
Metal analysis through AAS revealed presence of high amounts of zinc, copper, chromium, iron, arsenic and mercury in industrial effluent. Various organic pollutants such as chlorpyrifos, cucurbitacin-b and phthalates were identified by screening through GC-MS.
Microbiological evaluation of textile dyeing industry wastewater revealed a high bacterial load. Different bacteria isolated from wastewater such as Staphylococcus aureus, Pseudomonas aeruginosa, Corynebacterium xerosis, Bacillus megaterium, Staphyoloccus epidermidis and Micrococcus varians exhibited resistance to Cr and Cu salts and antibiotics to varying degree.
Ames test with/without enzyme activation and MTT assay showed strong association of industrial effluent with mutagenicity and cytotoxicity respectively. Bacterial reverse mutation assay revealed that the mutagenicity of textile dyeing industry wastewater decreased with increase in dilution of wastewater.
In-vitro comet assay revealed the evidence of high oxidative DNA damage induced by textile wastewater. Wastewater exhibited concentration dependent genotoxicity in sheep
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peripheral lymphocytes. When Wistar rats were exposed to industrial effluent in different dilutions for 60 days, then activities of total superoxide dismutase and catalase and hydrogen peroxide concentration were found to be significantly lower in kidney, liver and blood/ plasma of effluent exposed rats than control. Vitamin C at a dose of 50mg/Kg/day significantly reduced oxidative effects of effluent in rats. Industrial effluents may decrease activities of T-SOD and CAT and concentration of H2O2 in liver, kidney and blood/plasma of Wistar rats. Vitamin C may have a possible ameliorating effect on industrial effluent induced oxidative stress in Wistar rats.
Wastewater exposed rats exhibited necrosis of epithelial cells of nephron, pulmonary emphysema, and inflammation of the lungs, degradation and infiltration of cardiac myocytes, fibrosis of the liver, damage to the intestinal mucosa and sloughing off epithelial cells from the intestinal lumen.
This study concludes that untreated textile dyeing wastewater being a complex mixture of inorganic and organic pollutants may be highly eco-toxic and may contaminate of the environment via continuous release of various organic and inorganic pollutants. Availability: Items available for loan: UVAS Library [Call number: 2580-T] (1).
49.
Chemical Microbiological And Toxicological Evaluation Of Pharmaceutical Effluent Wastewater
by Ali Sharif (2011-VA-266) | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Prof. Dr. Aftab Ahmad Anjum .
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pharmaceutical effluent being a complex mixture of drugs and heavy metals may affect human health exhibiting a strong potential of mutagenicity, carcinogenicity, cytotoxicity and oxidative stress induction along with pathological changes in various organs of the body. The current study was focused to quantify the presence of heavy metals, detection of various drugs, determining the bacterial load along with isolation and identification of different bacteria and assessment of the mutagenic and genotoxic, cytotoxic and oxidative stress induction of pharmaceutical effluent wastewater when exposed to sheep lymphocytes, Salmonella typhimurium strains, cell lines and rats respectively.
Atomic absorption spectrophotometer was used to quantify heavy metals and showed the presence of arsenic, chromium, lead and iron in concentrations above the normal limits recommended by WHO and EPA. Gas Chromatograph mass spectrophotometer analysis shown the presence of digitoxin, lignocaine, caffeine and trimethoprim and various other organic pollutants.
Microbiological evaluation showed a high bacterial load in the pharmaceutical waste water. Several bacteria were also found in PEW in the presence of different drugs and heavy metals. Aeromonas sobria, Micrococcus varians, Staphyoloccus epidermidis, Staphylococcus aureus, Bacillus megaterium showed tolerance to potassium di chromate and copper sulphate and resistance to various antibiotic discs.
Ames assay revealed a strong mutagenic potential with and without the presence of metabolic activation mixtures. A concentration dependent effect was observed when samples were tested with increasing dilution factor.
MTT assay and comet assay also showed a concentration dependent effect. The BHK-21 cell line was used to evaluate cytotoxicity and cell viability decreased with increasing concentration of PEW. Sheep lymphocytes used in comet assay exhibited a concentration dependent DNA damage.
Different antioxidant enzymes were also evaluated. Rats were exposed to PEW at different concentrations and following 60 days oral exposure, rats were evaluated for the presence of total superoxide dismutase, catalase and hydrogen peroxide in kidney, liver and plasma. Exposure to Pharmaceutical waste water significantly decreased the (TSOD), (CAT) and (H2O2) levels in plasma, liver and kidney. Treatment with Vitamin E significantly ameliorated the levels of enzymes.
Exposed rats were also evaluated for any pathological changes. Coagulative necrosis of renal epithelial cells were observed along with severe degeneration and cellular swelling in hepatocytes of hepatic cord.
Availability: Items available for loan: UVAS Library [Call number: 2600-T] (1).
50.
Effects Of Physico-Chemical Properties Of Diluents On The Infectivity Titers Of Freez Dried Ppr Virus Vaccine
by Fariya Yaqub Baig (2009-VA-240) | Dr. Jawad Nazir | Prof Dr. Aftab Ahmad Anjum | Dr. Waseem Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: 6.1. Introduction.
Peste des Petits Ruminants (PPR) is a febrile viral disease of sheep and goats. The disease is
responsible for low productivity in small ruminants and great economic losses to the farmers in
Africa and Asia including Pakistan. There is no specific treatment for PPR and prevention is only
possible through the use of live attenuated vaccines. Being an enveloped virus, PPRV is heat
sensitive. Poor immunological responses have been observed after the vaccination of PPR.
Reasons may be disturbance in maintenance of cold chain, improper handling and route of
vaccination as well as reconstitution in inappropriate diluents. Physiochemical properties
(temperature, pH, osmotic pressure, salinity and UV light) of diluents effects the infectivity of
PPR freeze dried vaccine as live virus vaccines work properly after reconstitution within the
recommended time interval.
6.2. Experimental Design.
Effect of three diluents (normal saline, phosphate buffer saline and distilled water)
adjusted to various pH conditions (5.00, 6.00. 7.00, 8.00, and 9.00) on the infectivity of PPRV
was evaluated. Contents of the freeze dried PPR vaccine vials were diluted with one ml of the
respective diluent adjusted to above mentioned pH conditions. Virus infectivity from the vials
was measured immediately following reconstitution. One set of vials was kept at room
temperature (25 ºC ±2) and virus infectivity was measured afterwards at 30, 60, and 120 minutes
as described in the section 3.3.2. While other set of the vials was kept at refrigerated temperature
(4 ºC ±2) and virus infectivity was measured at 30, 60, 120, and 180 minutes as described in the
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46
section 3.3.2. The change in pH of each vial following reconstitution was also measured
accordingly. Each set of experiment was repeated three times independently.
6.3. Results.
PBS and NS gave equivalent results and proved better than distilled water to restore the
infectivity of PPRV. For a better comparison of virus stability in the PPR freeze dried vaccine
following reconstitution in various diluents adjusted to various pH conditions the infectivity titer
of the virus in the vaccines were measured at various time points. The serial data thus obtained
was analyzed by linear regression model to calculate T-90 values (Time required for 90 %
reduction in virus infectivity). The higher T-90 values indicate better stability of the virus at a
designated condition. At both of the temperatures the T-90 values were equivalent and relatively
higher for all of the diluents adjusted to pH 7.00 and 8.00 while these values are lower at extreme
pH conditions. Minimum T-90 values (126± 56) were observed at pH 9.00 in normal saline kept
at ambient temperature while maximum T-90 (448 ± 49.7) values were observed at pH 7.00 in
phosphate buffer saline kept at refrigerated temperature.
6.4. Conclusion.
Results of the present study suggest that virus infectivity in the live attenuated PPRV vaccine can
be better stabilized following reconstitution in PBS adjusted to neutral or slightly alkaline pH.
Chilled vaccine diluent is preferable to the one kept at room temperature and vaccine should be
administered within 30 minutes of reconstitution. Availability: Items available for loan: UVAS Library [Call number: 2645-T] (1).
