Detoxification Potential Of Yeast Sludhe Ahainst Ochratoxin In Broiler Chicks
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Publisher: 2011 Dissertation note: Ochratoxin the fungal secondary metabolite is a potent natural contaminant of poultry feed. Mycotoxins present in poultry feeds from the raw material used in their production is the major cause of toxic feed. The intake of very low levels of Ochratoxin-A result in overt ochratoxicosis resulting in impairment of immune system and acquired resistance to infections causing health problems which lead to economic losses in the form of reduced productivity The research study was conducted to study the harmful effects of Ochratoxin on broiler chicks and the adsorptive potential of yeast sludge against Ochratoxin in broiler chicks .
Aspergillus ochraceus was grown on Sabraud's Dextrose Agar and ochratoxin was produced on fermented wheat grains .One fifty day old Chicks of broiler breed were purchased from Big birds hatchery and were raised on commercial broiler diet till 7 days.
Four diets A,B,C and D were formulated A diet serve as control, B diet contained OTA 500ppb, C diet contained OTA 500ppb and 1% Yeast sludge and D diet contained OTA 500ppb and 2% Yeast sludge. These four diets were assigned randomly to the chicks, such that there were three replicates on each ration and each replicate contained 10 chicks. Vaccination against N.D and IBD was performed according to the schedule. During feeding trial weight gain , feed consumed, FCR and mortality rate was determined.
Group B (500ppb OTA) showed a decrease in weight gain and feed consumption as compared to group A (control diet) , C (1% yeast sludge and 500ppb OTA) and D (2% yeast sludge and 500ppb ochratoxin). Group D showed more improvement in weight gain, feed consumption and FCR as compared to group C.
Blood serum and tissue samples were collected from the birds slaughtered at the end of experimental trial. Concentration of serum total protein, albumin and activity of alanine transaminase were determined. Blood Serum levels of total protein and albumin were lower in the group B (500ppb OTA) than group D having 2 % yeast sludge but the group C fed on 1% yeast sludge did not show much improvement in those parameters. Activity of ALT was found to be significantly higher (P<0.05) in group C as compared to all other groups. Whereas blood serum ALT activity of the birds fed on ration B was significantly high (P< 0.05) as compared to blood serum ALT of group A
The Level of Ochratoxin in Liver and Kidneys was also determined and it was found to be highest in Group B (500ppb OTA) and lowest in Group D (500ppb OTA + 2% yeast sludge).
Based upon the observations obtained in this study it can be concluded that ochratoxin-A is a nephrotoxic and hepatotoxic agent. But supplementation of 2% yeast sludge in the broiler diet can effectively detoxify the effects of ochratoxin as compared to supplementation of 1% yeast sludge in the chicks diet.
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Antibacterial Activity Of Indihenous Hernal Exteacts Ahainst Urease Profucinh Bacreria
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Publisher: 2011 Dissertation note: Poultry farming is a profitable business but is facing serious ammonia environment particularly during winter season when ventilation frequency is reduced to maintain the shed's temperature. Urease producing bacteria in droppings are main cause of emitting ammonia in the sheds. The ammonia poultry environment is inducing reduced weight gain, immuno-suppression, enhanced susceptibility to respiratory pathogens, etc. Aqueous and alcoholic extracts of 14 local herbs (Aloe Vera, Azadirachta indica, Allium sativum, Calotropis procera, Cannabis sativa, Carum capticum, Eucalyptus camaldulensis, Lantana camara, Mangifera indica, Mentha piperita, Nigella sativa, Opuntia ficus indica, Piper nigrum and Zingiber officinalis) and four commercial herbal products (Mentofin, Suduri, Safi, Yucca) were evaluated for their in-vitro antibacterial activity against Proteus mirabilis by serial dilution method. It was observed that with reference to rise in pH, Ammonia concentration and urease activity in aqueous and alcoholic extracts of Allium sativum (pH: 8.5560, 8.8480, Ammonia:4.42, 3.52 µg/mL, Urease: 0.009, 0.007 U/mL respectively) had shown best results as compared to control positive (pH: 9.03, Ammonia: 6.7µg/mL, Urease: 0.013 U/mL). Alcoholic extracts of Mangifera indica (8.8820, 5.42µg/mL, 0.010 IU/mL), Mentha piperita (8.8880, 4µg/mL, 0.008 U/mL) Carum capticum (8.9540, 4.84µg/mL, 0.009 U/mL) and aqueous extract of Opuntia ficus indica (8.8100, 5.22µg/mL, 0.010 U/mL) had weak activity against P. mirabilis. Both aqueous and alcoholic extracts of Eucalyptus camaldulensis (pH: 8.91, 8.96, Ammonia: 5.16, 5.06 µg/mL, Urease: 0.01, 0.01 U/mL) has weak inhibitory effect. All commercial products had shown strong antibacterial activity (pH: 4.8-6.8, Ammonia: 0µg/mL, Urease: 0 U/mL). Results of remaining herbal extracts were not significantly different (p<0.05) from positive control. It was concluded that all herbal products had strong antibacterial activity against P. mirabilis. Mentofin had shown best results with optimum inhibitory concentration (1/1000 mL). Alcoholic extracts of few herbs had shown weak bactericidal activity. These herbs might give better results in-vivo.
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Suitability Of In-House Developed Pt-Pcr Fro The Detection And Serotyping Of Dengue Virus In Pakistan
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Publisher: 2013 Dissertation note: Dengue Virus (DENV) belongs to the genus Flavivirus of family Flaviviridae having four serological different serotypes such as DENV1, DENV2, DENV3 and DENV4 (Bai et al., 2008) Being a Flaviviridae member, the dengue virus is transmitted to human by genus Aedes, mainly Aedes agypti. Over the years dengue fever has become a significant infectious disease in different parts of the world that leads and increases the growth of mosquitoes. It has become epidemic in more than 100 countries on the globe with more than 2.5 billion people at the risk of infection. Pakistan has witnessed some severe outbreaks of dengue viral infection which results to major morbidity and mortality since mid of 90s. There is a need to overcome this infectious and in many cases fatal disease. Imprecise fatality morbidity and statistics underrate the magnitude of dengue as a regional health problem. Medical and public health services have been incapable to diminish this infection since there is no current vaccine available to prevent infectious disease, no effective medical treatments that avert the development of severe symptoms and no sustainable control measures against the vector that guarantee protection of affected communities.
Management of dengue patients and principally dengue hemorrhagic fever (DHF)/Dengue shock syndrome (DSS) cases are the alarming challenges now a day and in the upcoming episodes in this country. To deal with this challenge a sensitive and specific technique is required for its early diagnosis along with the knowledge of dengue serotype to increase the specificity of diagnosis and treatment. This study was designed to check the usefulness of nucleic acid based molecular determination of dengue virus along with nucleic acid sequencing/ analysis of different Dengue serotypes through phylogenetic studies.
Total 50 Blood samples were collected from the dengue suspected patients in 2011 outbreak of dengue. Samples were analyzed by PCR based detection and were compared with IgG, IgM detections to check the usefulness of PCR based nucleic acid detection. In second phase of study nucleic acid sequencing was done
The study has recommended PCR as a suitable and sensitive method for the rapid detection of dengue virus as it was found more sensitive than other utilized techniques including antibodies detection however it was not found useful to differentiate between primary and secondary infection for which a combination of IgG, IgM is more helpful choice. Nucleic acid analysis helped to define the common serotypes/genotypes of dengue virus circulating in Pakistan. In addition the present study has correlated our studied serotypes to other serotypes circulating in the globe which showed 98% homology with Srilankan strain and find out sequence similarities of our serotypes to the other serotypes distributed worldwide through phylogenetic analysis.
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Lysine Production On Pilot Scale By Brevibacterium Flavum And Its Characterization, Purification And Crystallization
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Publisher: 2013 Dissertation note: Food and feed protein demands have increased due to raise in population. Therefore continuous efforts have been progressed to enhance the production rate by conventional and non conventional methods. Fermentation technology have participated decisive role for a long time period and presently the amino acids formed by fermentation set apart principal biotechnology products significantly. By consuming low-cost carbon supply mutants originate potential to the inexpensive built-up for amino acids. L-lysine demand is steadily rising in the sector of feed stuffs, soft drinks, food ingredients, pharmacy and biological fluids, etc. In order to meet the market demand and accomplish growing and assorted L-lysine requirements, microbial metabolic engineering and recombinant DNA technology is the only hope and possibility for advancing the strains. Purification and isolation of material produced is a very significant element extremely influences fermentation practice usefulness and manufacturing expenses. It demands enhancement in the recycling procedure of amino acids, mainly L-lysine.
The present study was designed to produce lysine on pilot scale by using Brevibacterium flavum. A variety of agricultural byproducts like wheat bran, sugar cane molasses and rice polishing were utilized as substrate for lysine production through fermentation by using Brevibacterium flavum. Primarily optimum conditions were determined through fermentation for lysine production on micro scale. Subsequently these conditions were employed for biosynthesis of lysine on pilot scale. Qualitative assay of lysine was performed by TLC and quantitative assay by spectrophotometrically. It was found that amongst all the substrates 4% molasses was produced maximum lysine at 300C. Different inorganic and organic material like 0.4% CaCO3, O.4% MgSO4.7H2O, 0.1% NaCl, 0.8% KH2PO4, 2.5 % (NH4)2SO4, 0.5 % urea, 0.04 mg % biotin and 0.6 % corn steep liquor were found to be optimal for maximum lysine yield. After pilot scale production of lysine in fermentor, different techniques of downstream were applied. The biomass liquor thus produced was purified and crystallized through different techniques to transform in to L-lysine crystals. The information thus attained was subjected to statistical analysis by using one way ANOVA on optimization of different parameters for L-lysine production and comparison of mean values was done by Least Significant Difference (LSD).
Based on the above observations it was concluded that molasses is the most suitable substrate among other agriculture wastes for maximum lysine production with Brevibacterium flavum.
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Extraction, Purificaton And Characterization Of Proteolytic Enzyme From Fig (Ficus Carica)/ Karachi
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; Literary form:
Publisher: 2013 Dissertation note: Today, the enzymes are generally used in various industrial applications and require for more stable, highly active and specific enzymes are growing rapidly. Global market for industrial enzymes is reported to be €1 billion in 1995 (Godfrey and West, 1996) whereas, it was increased to $2.3 billion in 2007 and was expected to increase to over $2.7 billion by 2012.
In this piece of research work, purification and characterization of papain (a proteolytic enzyme) from Kachri (Cucumis trigonus) and Ficus (Ficus carica) were carried out. Extraction of papain was done using 0.1M alkaline phosphate buffer of pH 8.00, 70% ethanol and dist.water. Purification of papain was carried out by Ammonium Sulphate precipitation and dialysis followed by Gel filtration by Sephadex G-50. Then characterization of papain such as protein estimation, determination of proteolytic activity (international Unit) of enzyme and SDS-PAGE analysis were performed to determined molecular weight. Finally, the yield and proteolytic activity of papain was measured and compared with the commercial product available in the market.
Crude preparation of enzyme has a wide specificity due to the presence of various proteinase and peptidase isozymes. The performance of the enzyme depends on the plant source, the climatic conditions for growth, and the methods used in its extraction and purification, for example, if the fruit is healthy, then enzyme found is more active.
Papain is used in many industries such as breweries, pharmaceuticals, food, leather, cosmatics, detergents, meat and fish processing for a variety of processes. Therefore, the end use segments are many in signifying that papain has high export demand (Ezekiel and Florence, 2012).
In case, Kachri and Ficus contain high concentration of proteolytic enzyme. These enzymes being present in natural fruit were free from any toxic effect. Hence can be used in food and pharmaceutical industries.
Student's t-Test was used for comparing the means of two samples Kachri (Cucumis trigonus) and Ficus (Ficus carica).
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