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.
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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.
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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.
Material type: ; Format:
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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.
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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.
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9.
Physico-Chemical Factors Affection Survival Of Mycoplasma Gallisepticum
by Javed Muhammad | Dr. Aftab Ahmad Anjum | Prof. Dr. Khushi Muhammad.
Material type: ; Format:
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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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10.
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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11.
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.
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12.
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).
13.
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:
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; 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).
14.
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.
Material type: ; Format:
print
; Literary form:
not fiction
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.
Availability: Items available for loan: UVAS Library [Call number: 1686,T] (1).
15.
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.
Material type: ; Format:
print
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.
Availability: Items available for loan: UVAS Library [Call number: 1687,T] (1).
16.
Antibody Response Of Buffalo Calves To Oil Based Multivalent (Pasteurella Multocida, Clostridium Chauvoei And FMD Virus "O' "A" and "Asia1") Vaccine
by Muhammad Farooq | Prof. Dr. Khushi Muhammad | Dr. Aftab Ahmad Anjum | Prof. Dr.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Microbial diseases are one of the constraints for further development of dairy industry as a profitable enterprise. The diseases are causing heavy economic losses to the industry. The diseases such as Foot and Mouth Disease (FMD), Hemorrhagic Septicemia (HS), Black Quarter (BQ),etc., are endemic in Pakistan and perpetuate among the dairy animals. These diseases can be effectively controlled by vaccination.
FMD virus "O", "A" and "Asia 1" were grown on BHK-21 and were inactivated with BEI. Culture of P. multocida and Cl. chauvoeiwere grown on CSY and RCM media, respectively and inactivated with formalin. The vaccine containing 0.2 x 107 units of TCID50of each serotype of FMD virus ("O", "A" and "Asia1"), 2 mg of Pasteurella multocida and 250 Hemolytic units of Clostridium chauvoei per dose were prepared. Oil adjuvanted vaccines of HS, HS + BQ, HS + FMD ("O", "A" and "Asia 1"), BQ, BQ+ FMD ("O", "A" and "Asia 1"), FMD ("O", "A" and "Asia 1") and HS + BQ + FMD ("O", "A" and "Asia 1") were prepared and injected into the buffalo calves in 7 group of 3(n=3) animals each separately at Living Dairies, Chunian. 8th group of three animals was kept as negative control. Antibody response against FMD virus, Cl. chauvoei and P. multocida were measured by CFT, Anti hemolytic Assay and IHA, respectively at day 0, 30, 60 and 90 post vaccinations. Two groups (n=3) of calves vaccinated with whole culture FMD vaccine and NSP free FMD vaccine. Data was analyzed by one way ANOVA procedure and significance was determined by Duncan Multiple Range Test through SPSS version 13.
The vaccine when injected in buffalo calves induced Log22.00±1.00units of anti FMD "O" CFT antibody titer, Log22.22±1.00 units of anti FMD "A" CFT antibody titer, Log22.22±0.84 units of anti FMD "Asia 1" CFT antibody titer; Log22.99±0.58 units of Indirect Haemagglutinating (IHA) units of antibody against Pasteurella multocida and Log25.44±1.02, Anti Hemolytic Units (AHU) of the antibodies against hemolytic toxins of Clostridium chauvoei. There was no significant difference among the titers of FMDV "O", "A" and "Asia 1"; Pasteurella multocida and Clostridium chauvoei whether used in monovalent or in multivalent.In present study anti-NSP-FMD virus ELISA antibodies in the animals vaccinated with FMD (whole culture) vaccine were undetectable on 15 days post priming while detectable on 30 and 45 days post priming. However anti-NSP-FMD virus ELISA antibodies in the animals vaccinated with FMD (NSP free) vaccine were undetectable on 15, 30 and 45 days post priming. Moreover these antibodies were detectable in FMD carrier animals on 15 days post recovery.Cellular pellet of Pasteurella multocida, Clostridium chauvoei can be used to further minimizing the volume of culture required and further Brucella abortis vaccine can be added in it in conjunction with FMD. This will revolutionize the field of vaccination in Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 1732,T] (1).
17.
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.
Material type: ; Format:
print
; Literary form:
drama
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).
18.
Identification Of Multiple Drug Resistant (Mrd) Mastitis Causing Bacteria In Dairy Goats
by Muhammad Faisal najees | Dr. Aftab ahmad anjum | Prof. Dr. mansur-ud-Din ahmad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1852,T] (1).
19.
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).
20.
Plasmid Mediated Analyses And Plasmid Curing Of Previously Isolatedmulti-Drug Resistant Eschetichia Coli From Retail
by Mawra gohar | Dr. Ali ahmad sheikh | Dr.Tanveer | Prof, Dr. Aftab ahmad anjum.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1923,T] (1).
21.
Isolation Characterization And Optimization Of Potential Probiotic Bacteria From Poultry Droopings
by Muhammad Hashim khan | Prof. Dr. Aftab ahmad anjum | Dr. Jawad nazir | Prof. Dr. Mansur-ud-din.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1991,T] (1).
22.
Isolation And Characterization Of Antibiotic Resistant Lactobacilli From Fermented Food Products
by Shahgull | Dr. Muhammad Nawaz | Prof. Dr | Prof. Dr. Aftab anjum.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2002,T] (1).
23.
Prevelance Of Brucellosis In Aborted Women Visiting Tertiary Care Hospitals Of Lahore City
by Saba Yasmin (2009-VA-211) | Prof. Dr. Aftab Ahmad Anjum | Dr. Tayyaba Ijaz (Co Supervisor) | Prof. Dr. Khushi Muhammad | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pakistan is an agriculture based country whose rural population depends upon livestock for livelihood. Contribution of livestock to agriculture sector is 55.9 percent while 11.8 percent to the national GDP during 2013-14 (GOP 2013-2014). A number of infectious diseases hamper the growth of livestock sector. Some of the livestock diseases are zoonotic in nature and threat to human health. Brucellosis is considered among major zoonotic diseases throughout the world. The Mediterranean Basin, south and Central America, Eastern Europe, Asia, Africa, the Caribbean and the Middle East are considered as high-risk countries (Memish 2001).
Brucellosis in human beings is a major concern of community health. It causes acute and chronic illness, physical incapacity and loss of health. Bacterial species involved include Brucella abortus, Brucella melitensis or Brucella suis. Brucellosis is acquired by human beings from infected animals by close contact with vaginal secretions, urine, feces, blood, aborted fetus, or consumption of unpasteurized milk or other raw milk products. Shepherds, milkmen, butchers, knackers, veterinary assistants and abattoir workers are at high risk (Agasthya et al. 2007). Prevalence of brucellosis recorded by Mukhtar and Kokab (2008) in abattoir workers of Lahore Pakistan was 21.7 percent. Higher prevalence of brucellosis was observed in females (37.06%) than males (24.2%) in patients admitted at Peshawar, Pakistan (Shahid et al. 2014).
Symptoms of disease vary among human patients, ranging from non–specific, flu-like symptoms (acute form) to undulant fever (chronic form). Some of the serious complications of skeletal system, cardiovascular and central nervous systems may develop. Other important signs observed include arthritis, orchitis, epididymitis, abortion, retained placenta and stillbirth (Baba et al. 2001; Grilló et al. 2006). In animals, brucellosis in most of the cases results in abortion, birth of weak calves, death of young stock, infertility in males and reduced milk yield in females (Maadi et al. 2011; Abubakar et al. 2012).
There is actual need for teamwork between public health officials and veterinary officers to reduce communication of brucellosis between animals and human in endemic areas (Jelastopulu et al. 2008; Makis et al. 2008). Clinical picture of brucellosis is nonspecific and may vary from patient to patient. Therefore, laboratory diagnosis by isolation and culture or recognition of specific anti–Brucella antibodies is essential for confirmation of brucellosis (Al-Attas et al. 2000).
Diagnosis of brucellosis by culture and phenotypic description is time-consuming. Furthermore, risk of infection to worker is always there. Serological tests are commonly preferred for brucellosis in cattle and small ruminants, especially at farm level screening. Chance of cross-reactions with other gram negative bacteria is a major problem. Rose Bengal Plate Agglutination Test (RBPT) and Slow Agglutination Test (SAT) are extensively used for detection of anti-Brucella antibodies (Halling et al. 2005). Enzyme Linked Immunosorbent Assays (ELISA) have been developed to resolve suspected samples by RBPT. ELISA is more sensitive, so it can detect Brucella carriers which are negative by RBT, SAT and CFT (Aert et al. 1984). Molecular techniques are more reliable and specific than serological tests. Final confirmation of brucellosis is carried out using polymerase chain reaction (PCR), a molecular technique. Real-time PCR offers enhanced sensitivity, specificity and rapidity of performance when compared to conventional PCR (Gwida et al. 2012). Availability: Items available for loan: UVAS Library [Call number: 2225-T] (1).
24.
Modulation Of Antibiotics Resistance Pattern In Escherichia Coli By Different Plant
by Bushra Chaudary (2009-VA-232) | Dr.Muhammad Nawaz | Prof. Dr. Aftab Ahmed | Dr. Naureen Naeem.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Escherichia coli (E. coli) is Gram negative microorganism belonging to family
Enterobacteriaceae. It is part of normal micro flora of gastrointestinal tract of human and all
warm blooded animals (Kaper et al. 2004). Escherichia coli is source of many infectious diseases
in human as well as in animals. Common E. coli infections are enteritis, urinary tract infection,
septicemia and neonatal meningitis. In pets and farm animals, E. coli is associated with diarrhea
(Allocati et al. 2013). Poultry industry is facing huge annual losses due to infection of avian
Pathogenic E. coli (APEC) in broilers (Oosterik et al. 2014). E. coli causes a variety of
syndromes in poultry including yolk sac infection, respiratory tract infection, swollen head
syndrome, septicemia and cellulitis (Buys et al. 1989)
Antibiotics are chemical agents which inhibit the microbial growth and used to eradicate
infections. Mechanisms of action of antibiotics provide a base to categorize antimicrobial agents.
Most important classes of antibiotics act as inhibitors of cell wall synthesis, protein synthesis
(tetracyclines and macrolides), nucleic acid synthesis (fluoroquinolones), metabolic pathway
(trimethoprim-sulfamethoxazole) and cell membrane (polymyxins). Bacteria may have intrinsic
or acquired resistance to antimicrobials (Tenover 2006).
Urinary tract infections are mostly caused by E.coli. Antibiotics generally used for the treatment
of E. coli infections include ampicillin, nitrofurntion, cephalosporin, sulphonamides
(trimethoprim-sulfamethoxazole) and quonolones (neladixic acid, ofloxacine, ciprofloxacin and
levofloxacine) (Lin and Lin 2010).
Extended use and misuse of antibiotics lead to the development of resistant bacteria. Resistant
E. coli strains are common source of hospital born and community acquired infections. Ease of
Introduction
2
international travelling is one of the major spreading factor for antibiotic resistance. Resistant
bacteria got opportunity to move from one geographical area to another (van der Bij and Pitout
2012). New strains of E. coli resistant to carbapenems (New Delhi metallo-β-lactamase 1 (NDM-
1) are major global health issue (Kumarasamy et al. 2010). Antibiotic resistance has become a
serious public health problem. Currently, world is facing great difficulty in treatment of many
infectious disease of human and animals. One of the reasons of treatment failure is emergence of
resistant bacteria (Levy 2002).
To develop new strategies for treatment of infectious diseases, it is necessary to understand the
mechanisms of resistance. Efflux pump inhibitors, enzymatic degradations and alteration of
target sites are major strategies by which bacteria acquire or develop resistance to antibiotics
(Sibanda and Okoh 2007).
Scientists are looking for alternatives of antibiotics such as bacteriopheges, naturally
antimicrobial compounds and some non antimicrobial agents (Worthington and Melander 2013).
Probiotics (Lactobacillus and bifidobacterium) can be a prophylactic measures against E. coli
and may be used to treat intestinal tract infections of E. coli and other bacteria (de Vrese and
Schrezenmeir 2008).
Phytochemicals, secondary metabolites of plants, have antibacterial activity against many
pathogenic organisms. These phytochemicals in combination with antibiotics may show
synergistic effect. Phytochemicals and plant extracts can be a source of antibiotic resistancemodifying
agents (RMAs) (Abreu et al. 2012). Plant extracts shown antibacterial activity
because of phytochemicals like alkaloids, tannins, flavonoids, phenolic compounds and steroids
(Gobalakrishnan et al. 2013). Plant extracts are used as traditional medicine for the treatment of
many diseases. Plant extracts like Zingiber officinalis (Ginger) Gymnema sylvestre (Gurmar buti), Astragalus (goat’s thorn), Calotropis procera (apple of Sodom) and oputia dillenii (cactus)
have antimicrobial activity (indu et al. 2006 and Kumaar et al. 2013). Plant extracts also have
antibiotic resistance modulation potential (Mako et al. 2012).
Availability: Items available for loan: UVAS Library [Call number: 2247-T] (1).
25.
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).
26.
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).
27.
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).
28.
Sero-Screening Of Camels For Different Infectious Diseases
by Mazia Khalid (2008-VA-358) | Dr. Aamir Ghafoor | Prof. Dr. Aftab Ahmed Anjum | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Camel is the precious and important animal in Pakistan. Camel is the most well adapted livestock species, survives and produces in climatic extremes and is well appreciated for its significance in the pastoral economy of the province. The camel being an important livestock species uniquely adapted to hot and arid environments and therefore contributes significantly to the food security of the nomadic pastoral households. Although camel being hardiest animal is less susceptible to diseases as compared to other livestock animals but literature shows that some diseases are still prevalent in camels. In view of the significance of camel as livestock animal as well as the symbol of cultural heritage of the nomadic pastoralists, there is a need to combat different diseases to which camels are susceptible and then appropriate control strategies should be applied.
Present study was designed to check the percentage positivity of different major diseases in camels that may pose serious issue relating to camel health and its importance as an important livestock animal. The diseases included in this study are Q fever, Brucellosis, FMD, CBPP and Neosporosis. ELISA is used to detect antibody prevalence by using specific kit based protocol for each disease whereas in case of Brucellosis RBPT is also used as basic screening test.
And it was found that Q fever has highest percentage seropositivity in both districts as compared to other diseases whose presence in camels was found to be almost seronegative.
So it was concluded that camel is still resistant to many diseases though some diseases are still prevalent in camels and these diseases should be controlled through public awareness and routine screening. Availability: Items available for loan: UVAS Library [Call number: 2401-T] (1).
29.
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).
30.
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).
31.
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).
32.
Antibiotic Resistance Pattern Of Staphlococcus Aureus And Its Resistance Modulation Using Medicinal Plant Extracts
by Iqra Asif (2010-VA-279) | Prof. Dr. Aftab Ahmed Anjum | Prof. Dr. Khushi Muhammad | Ms. Tehreem Hussain.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This project was designed to evaluate the antimicrobial efficacy of Chloroform and
ethanol extracts of Calotropis procera (C .procera) and Eucalyptus globulus (E. globulus)
against Multiple Drug Resistant (MDR) Staphylococcus aureus isolated from human origin. This
study was conducted to evaluate the antimicrobial potential of C. procera and E. globulus alone
and in combination with antibiotics to check synergism between medicinal plants and resistant
antibiotics.
S.aureus is a major pathogen which causes various infections. Infectious diseases affect
millions of people around the world and in the history these diseases are major cause of mortality
and morbidity across the globe. In past few decades rate of mortalities are continuously
increasing because of acquired resistance of S. aureus against multiple drugs, thus it is utter need
of time to discover some alternatives to antibiotics so that we can resolve this dilemma of
antibiotic resistance. Plant extracts are hope for this purpose as they have many compounds
which have potential to lower down the number of micro-organisms. Plants have benefits over
other as they are non toxic, non-reactive and have least side effects.
Total 20 samples of human origins were procured from Department of Microbiology,
UVAS Lahore and were subjected to check their antibiotic resistance profile against
Erythromycin, Amoxicillin and Ciprofloxacin by Kirby Bauer disc diffusion assay. Out of 20,
nine resistant isolates were separated. Among them three were resistant to Erythromycin, three to
Amoxicillin and three to Ciprofloxacin.
Summary
72
Leaves of C. procera and E. globulus were processed in Chloroform and ethanol
Solvents. Antimicrobial activity was evaluated by agar gel well diffusion assay in which zone of
inhibitions were measured. Minimum inhibitory concentration (MIC) of plant extracts was
evaluated by micro broth dilution method. Best antimicrobial activity was observed by ethanolic
extract of E. globulus.
Then combine effect of sub-inhibitory concentrations (SICs) of plant extracts and
minimum inhibitory concentration of antibiotics were determined by Well Diffusion assay. Four
different sub inhibitory concentrations of plant extracts i.e.10μg/ml, 20μg/ml, 40μg/ml and
80μg/ml were used in combination with fixed concentration of antibiotics i.e.100μg/ml to check
combinational effect of both. At selected sub-inhibitory concentration plant extract alone did not
show any antibacterial activity. Two of the isolates had shown modulation when amoxicillin and
plant extracts combination was used against them. The isolate labeled as S.aureus 4 showed
modulation with the use of Ethanolic extract of Calotropis procera and S.aureus 5 had shown
modulation with the use of chloroform extract of Calotropis procera. For further confirmation
two more concentrations of 160μg/ml and 320μg/ml were used along with100μg/ml Amoxicillin
against same isolates S.aureus 4 and 5. Zone of inhibition was observed with increased diameter
indicating modulation of two isolates. While erythromycin and ciprofloxacin resistant isolates
didn’t show any modulation.
Data of antibiotic resistance and resistance modulation using plant extracts was analyzed
by one way analysis of variance (ANOVA) followed by Duncan’s multiple range(DMR)
posthoc test using Statistical package for social sciences (SPSS) 17.0 Statistical software at α <
0.05. Availability: Items available for loan: UVAS Library [Call number: 2501-T] (1).
33.
Antibacterial Activity Of Plant Extracts Against Antibiotic Resistant Pseudomonas Aeruginosa And Their Cytotoxicity Profile
by Hafiza Farah Asghar (2010-VA-276) | Prof. Dr. Aftab Ahmed Anjum | Dr. Ali Ahmad Sheikh | Muhammad Nasir.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2016Dissertation note: Pseudomonas aeruginosa is a common opportunistic pathogen of respiratory tract
and cause both hospital and community acquired infections. For the treatment of
infections antibiotics are used but due to random selection of commonly used antibiotics,
resistance in P. aeruginosa has developed. This problem may leads to the high morbidity
and mortality rate. Different medicinal plants have antibacterial activity in their
secondary metabolite. Secondary metabolites are terpens, flavonoids, alkaloids and
alcohols etc. So the plant extract could be the alternative therapy for the treatment to
reduce the antibiotic resistance problem.
Isolates of P. aeruginosa was procured from the main clinical laboratory of Mayo
Hospital, Lahore and identified biochemically according to bergey’s manual of
determinative bacteriology. Antibiotic resistance pattern of identified P. aeruginosa was
evaluated by Kirby Bauer disc diffusion assay against selected antibiotics includes
ciprofloxacin, levofloxacin, meropenem and imipenem. Measure the zone of inhibition
and isolates marked as resistant, intermediate and sensitive. Resistant strains were
alienated for further evaluation.
Leaves of Eucalyptus globulus (Tasmanian blue gum) and Calotropis procera
(apple of Sodom) proceed for extraction and the plant extracts was obtained by using
solvent chloroform and ethanol. Percentage yield of both plant extract was calculated.
High percentage yield was obtained from Eucalyptus globulus and less percentage yield
was gained from Calotropis procera in comparison The obtained extract was dried and
the resultant material was used in well diffusion assay to evaluate the antibiotic
CHAPTER 6
SUMMARY
Summary
66
sensitivity of resistant P. aeruginosa against selected plants. Stock of plant extracts was
prepared by dissolving 1g of plant extract in 1ml of DMSO. Well diffusion assay was
performed and zones were measured in millimeter and categorized as resistant, sensitive
and intermediate. Isolates that are susceptible to plant extracts were separated and
Minimum inhibitory concentration of susceptible isolates was determined by broth micro
dilution assay and cytotoxicity profiling was done by 3-(4, 5-dimethyl-2-thiazolyl)-2,5-
diphenyl-2H-tetrazolium bromide (MTT) assay and cell survival percentage was
calculated.
Data recorded during the study was analyzed by one-way analysis of variance
(ANOVA) followed by Duncan’s test using the SPSS statistical software program.
Differences were considered significant at P < 0.05. Availability: Items available for loan: UVAS Library [Call number: 2545-T] (1).
34.
Comparative Quality Evaluation Of Raw And Pasteurized Milk
by Hafiza Saima Ghaffar (2009-VA-230) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmed Anjum | Dr. Sana Ullah Iqbal.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This particular project was designed to evaluate the overall quality of raw and pasteurized milk available at different areas of Lahore. The parameter which was checked includes microbiological analysis, adulterants, physicochemical properties and the effect of temperature on vitamin C in milk samples. Raw samples were collected from ten different towns of Lahore, whereas pasteurized milk samples belong to ten different brands. Ten samples were collected under control conditions from animals in sterilized containers.
For microbiological analysis four parameters were selected including total plate count (TPC), total coliform count (TCC), total psychrotrophic count (TPSC) and total yeast and mold count (TYMC) whereas, different adulterants like adulteration test was done such as urea, starch, hydrogen peroxide, detergent or soap, sorbitol, quaternary ammonium compound, boric acid, cane sugar, sodium chloride, formalin and hypochlorite were checked by using the milk adulteration kit in QOL. Milk contains casein and whey proteins. Whey protein being added in the milk to increase its density which is considers being an adulterant. In this project whey protein was estimated in all milk samples by titration method. Physicochemical characteristics of milk are an important parameter to judge the quality of milk. These physicochemical properties include fat%, SNF%, density kg/m3, lactose%, solid/ash, protein% and pH. Physicochemical properties were evaluated mechanically by Milkoscan. Heat treatment is an important method to reduce the microbiological contamination of milk. These treatments may include pasteurization and UHT etc. During the heat treatment some of the micronutrients may deteriorate thus compromising the quality of milk. Vitamin C is among those heat labile micronutrients. Vitamin C was checked quantitatively in market and self-collected samples by using titration method.
It was concluded that total plate count TPC, TCC, TPSC and TYMC of raw milk samples were above the standard value indicating the poor quality of the milk. As far as the pasteurized milk samples were concerned ninety percent of the samples showing higher values for TCC, TPSC and TYMC. Total plate counts of all self-collected raw milk from a healthy animal were found within the standard value. Counts were in range of 3.8x 103 – 8.9x103 CFU/mL of all milk samples. TPC of all self-collected raw milk from a healthy animal were found within the standard TCC were found within permissible value (102 CFU/mL .TPSC were negative for all milk samples. TYMC were in range of 2.6x101 -7.2x101 CFU/mL. Among milk samples (n=10), three samples (30%) were positive for TYMC were while remaining samples (70%) were negative and showed no growth.
Physicochemical factor show that 50 percent of raw milk have low nutritional value as compared to the standards which are buffalo and cow milk contains 7.6, 4.5% fat, 3.8, 3.8 % protein, 5.1, 4.9% lactose, 0.78, 0.72% ash and 17.0, 13.9% total solid respectively. In raw milk mean of fat (%), solid not fat (%), lactose (%), Solid/ash (0%), protein(%) and pH were 4.50±0.03, 7.915±0.06, 23.05±0.055, 3.893±0.06, 3.85±0.05, and6.9±0.0.02 respectively. In pasteurized milk mean value for fat, SNF, lactose, ash, protein and pH were 3.48 ±0.13, 7.24±0.10, 3.60±0.05,0.5 ±0.06, 2.82±0.05, 7.2±0.20 respectively. Pasteurized milk is good for consumption.
Different adulterant such as urea, starch, hydrogen peroxide, Sorbitol, QAC, Boric acid, Cane sugar, NaCl, Carbonate, Formalin, hypochlorite, whey protein, Added water and soap /detergents were evaluated in all milk samples. Among these adulterant water (66%) was found in majority of milk samples, followed by whey protein (15%), starch (13%), (10%) NaCl and
(8%) cane sugar were detected in raw milk samples. n Pasteurized milk samples only added water (49%) and whey protein (31%) was detected.
Among the raw milk samples the maximum and minimum concentration of vitamin C was observed 0.33±0.02 and 3.33 ±0.02 mg/100ml and for pasteurized milk maximum and minimum concentration of vitamin C was observed 2.54mg/100ml and 0.32 ±0.02 mg/100ml respectively. In self- collected samples the minimum and maximum concentration of vitamin C was observed 5.25±0.02 and 8.34 ±0.04 mg/100ml respectively and after pasteurization in laboratory minimum and maximum concentration of vitamin C was observed 3.48±0.04 and 5.83 ±0.02 mg/100ml respectively. These observations had showed that pasteurization treatment decreased Vitamin C quantity. Availability: Items available for loan: UVAS Library [Call number: 2536-T] (1).
35.
Isolation, Molecular Identification And Antibiotic Resistance Pattern Of Salmonella Enterica From Fancy Birds
by Aqeela Kousar (2010-VA-303) | Mr. Muhammad Asad Ali | Prof. Dr. Aftab Ahmed Anjum | Prof. Dr. Mansur-ud-Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Salmonellosis is a disease with serious health issues related to food borne illness and
most of world’s population is suffering from it. Early diagnosis in case is very important for
treatment of disease. Salmonellosis may hidden as a carrier state, acts as zoonotic components
for transmission of disease. Therefore the test with more diagnostic value needs to be developed
like Polymerase chain reaction after culturing and microbiological examination.Salmonella
enterica infections continue to pose a significant risk for poultry industry and fancy birds.
Salmonella infections have been controlled by antibiotics but in recent times antibiotic resistance
in microorganisms especially in Salmonella is a global health issue. Antibiotic resistant
Salmonella has further compounded the problem. Poultry isolate of Salmonella enterica (n=150)
were procured from Jallo park, Safari park and household pets which are taken to Pet Centre
University of Veterinary and Animal Sciences Lahore then brought to Department of
microbiology UVAS Lahore and identified by biochemical testing, morphology, staining
characters and genus specific PCR. Antibiotic Susceptibility was checked by disc diffusion
method against amoxicillin (30μg), ampicillin (10μg), cefixime (5μg), , ceftazidime (30μg),
ceftriaxone (30μg), ciprofloxacin (5μg), gentamicin (10μg), nalidixic acid and tetracycline
(30μg) and resistant pattern was 100 % in ampicillin and tetracycline and 41.18% and 58.82% %
in gentamicin and ciprofloxacin respectively while antibiotic show 0% resistance. Fancy birds
are carriers of drug resistant Salmonellae.
A total of 150 samples collected from Zoo Lahore, safari park and household pet fancy
birds each of n=50. Samples will enriched by non-selective and selective media, After isolation
on selective media macroscopic, biochemical analysis and microscopic examination done. DNA
Summary
53
extracted from culture isolated from cloacal swabs and polymerase chain reaction performed
using primers. Amplication will be observed using Agarose gel electrophoresis.
Research highlighted the prevalence of Salmonella in fancy birds and its possibility of
transmission to human beings. Research also provided data on antibiotic resistance in
Salmonellae from fancy birds and its possible role in ever increasing problem of antibiotic
resistance. Availability: Items available for loan: UVAS Library [Call number: 2615-T] (1).
