1.
Serorrevalence Of Toxoplasmosis In Captive Birds
by Sadia ibrahim | Prf.Dr. Azhar Maqbool | Dr. Aftab | Dr. Muhmmad latif | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2010Dissertation note: Toxoplasmosis is caused by Toxoplasma gondii, an intracellular parasite which infects humans as well as wide variety of mammals and birds. Toxoplasmosis is found throughout the world and tends to be more prevalent in tropical climates. It is considered as one of the major public health disease. Risk of infection from turkeys, pigeons, ducks and quails to man is scarce, as they are intermediate hosts and are frequently infected by eating feed contaminated by cat faeces containing oocysts of T.gondii. The high seroprevalence in them could be related to large populations of homeless cats in the city and may suggests the significant role of urban pigeons in the epidemiology of toxoplasmosis.
A total of 200 serum samples were analyzed for anti-toxoplasma antibodies, of these 50 samples from ducks, 50 sampls from turkeys, 50 samples from pigeons and 50 samples from quails were collected at random. Under aseptic measures, 1-2 ml blood were withdrawn by venipuncture. All the serum samples wiere analyzed for specific IgG anti toxoplasma antibodies using Latex Agglutination Test (LAT). Positive reaction were visualized by naked eyes or magnifying hand lense.Commercially available Toxoplasma latex kit was used to determine the serum antibody. Results were interpretated. Data obtained was analyzed statistically by calculating the per cent sero-positivity of Toxoplasma gondii in ducks, turkeys, pigeons and quails.
In the present study the overall seroprevalence of T.gondii infection in pigeons, ducks, turkeys and quails is 8%, 12%, 16% and 4% respectively by using Latex Agglutination Test. In pigeons seroprevalence of T.gondii was 8% out of 50 pigeons examined one gave an antibody titer of 1:256, one was positive at 1:128, whereas two at 1:16 secreening dilution. In ducks seroprevalence of Toxoplasma gondii is 12% out of 50 ducks examined one gave an antibody titer of 1:256, two was positive at 1:128, whereas three at 1:16 secreening dilution. In turkeys seroprevalence of Toxoplasma gondii is 16% out of 50 turkeys examined three gave an antibody titer of 1:256, two was positive at 1:128, whereas three at 1:16 secreening dilution. In quails seroprevalence of Toxoplasma gondii is 12% out of 50 quails examined zero gave an antibody titer of 1:256, one was positive at 1:128, whereas one at 1:16 secreening dilution.
Availability: Items available for loan: UVAS Library [Call number: 1160,T] (1).
2.
Chemical Equivalence Of Different Brands Of Amoxicillin Trihydrate And Its Minimum Inhibitory Concentration
by Rana Adnan Ali | Prof.Dr.Muhammad Ashraf | Dr aftab Ahmad | Dr.Muhammad Adil Rasheed.
Material type: ; Format:
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Publisher: 2011Dissertation note: This project was designed to study the chemical equivalence of different brands of amoxicillin trihydrate (long acting and short acting) approved by the ministry of health and available in the market for veterinary use. Amoxicillin was measured by HPLC method developed and standardized in the laboratory. Limit of detection (LOD) and limit of quantification (LOQ) of the amoxicillin trihydrate was determined. Solutions of different concentrations were prepared from amoxicillin trihydrate reference standard for the determination of LOD. and were protected from light and stored at 2-8 oC until used. The LOD calculated by us was 0.100 (µg / ml) and LOQ was 0.5 (µg / ml). Correlation Coefficient should be ? 0.99 and the result obtained by the data was 0.99984050. Chemical equivalence of all brands was determined by using HPLC systems (Shimadzu & Agilent). Concentrations for reference standard (50, 25 and 10 ?g /ml ) and for each brand (Alomox LA, Amovet LA, Farmox LA, Novamox LA, Trioxyl LA, Amoxi-vet, Colimox, and Colimoxin) were used. All the results obtained showed that maximum percentage of assay obtained among long acting was of the brand Farmox LA (101 %) and in case of short acting was of Amoxi-vet (101%). Minimum percentage of assay among long acting was of brand Amovet LA (92 %) and in case of short acting was of Colimox (96%). MIC of amoxicillin against E.coli and Staphylococcus was determined by micro broth dilution test. According to our results 73.33 % E.coli were susceptible and 26.67% were resistant to the amoxicillin trihydrate. Our results showed that 86.67% Staphylococcus were susceptible and 13.33% were resistant to Amoxicillin Trihydrate (Reference Standard). It showed that this antibiotic is still very effective against the diseases produced by the Escherichia.coli and Staphylococcus aureus.
Availability: Items available for loan: UVAS Library [Call number: 1249,T] (1).
3.
Effect Of Various Concen Trayious Of Hydrogen Pereoxide On Chemical And Microbiogical Quality Of Raw Buffalo Milk
by Muhammad Ilyas Alam | Prof. Dr. Muhammad Ayaz | Dr. Aftab Ahmed Anjum | Dr. Imran Javed.
Material type: ; Format:
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Publisher: 2011Dissertation note: Milk is a complex mixture of fat, proteins, carbohydrates, minerals, vitamins and other miscellaneous constituents dispersed in water. Milk production in flush season is much more than in the normal. Milk production and supply fluctuate through out the year and during winter it is surplus to its demand. Surplus milk is available in winter due to new calving, less consumption of milk by the consumer. In winter season ample amount of green fodder is available to the animals which in turn increase the milk production.
Milk and milk products being very delicate and perishable food require special handling prior to the consumption and further treatment. Pakistan due to its harsh climatic conditions people are using different methods, for the preservation of milk. They are using different chemicals, additives and antibiotics to enhance the keeping quality of milk. Present study was planned to investigate the various concentration of hydrogen peroxide or raw buffalo milk and its effect on chemical and microbiological quality of raw buffalo milk.
Raw buffalo milk samples were collected from Dairy Animal Training and Research Centre, University of Veterinary and Animal Sciences, Ravi campus Pattoki Fifty samples of raw buffalo milk (100ml each) were collected to studied the nutritional composition and microbiological quality of the milk after adding the hydrogen peroxide. The hydrogen peroxide of different concentration i.e. 0.025%, 0.05%, 0.075%and 0.1% were used in this study. There was no significant change in the result regarding various nutritional composition of raw buffalo milk after adding the various concentrations of hydrogen peroxide. There is a slight change in the lactose % during study of 48 h storage of milk at different temperature. Statistically the change which occurred in lactose during storage is significant whereas over all decrease in Solid Not Fat is non significant
Mean value of TPC of raw buffalo milk treated with different concentrations of hydrogen peroxide storage at the three different temperatures indicated that at 10° C TPC was very less as compared to control. TPC at 30° C after 48 h was 9.83x106.Which was very less as compared to TPC of control i.e. 1.195 x107.
The effect of H2O2 on the quality of the milk is negligible as compared to the losses suffered without it. The hydrogen peroxide definitely have its effect as a preservative.. The use of preservative in milk and dairy products are not new in the countries where ambient temperature remains quite high. Our study suggests that the concentration of hydrogen peroxide to be used for the preservation of raw milk is 0.05 % to 0.1 %
Availability: Items available for loan: UVAS Library [Call number: 1291,T] (1).
4.
Chemical Equivalence Of Different Brands Of Oxytertacycline Hydrochloride And Its Minimum
by Sadaf Hina | Prof. Dr. Muhammad Ashraf | Dr. Aftab | Dr. Muhammad Adil Rasheed.
Material type: ; Format:
print
; Nature of contents: ; Literary form: Publisher: 2012Dissertation note: This project was designed to study the chemical equivalence of various brands of Oxytetracycline hydrochloride (long acting, short acting & PVP) approved by the ministry of health and available in the local market for veterinary use. Oxytetracycline was measured by HPLC method developed and standardized in the laboratory. Limit of detection (LOD) and limit of quantification (LOQ) of the Oxytetracycline by HPLC assay method were determined. From stock solution of working standard (Oxytetracycline hydrochloride) different concentrations 0.05, 0.1, 0.5, 1.0, 10, 25, 50 and 100µg per ml were prepared for the determination of LOD. The LOD calculated was 0.100(µg/ml) and LOQ was 0.5 (µg/ml). Correlation coefficient was 0.99994050. Concentration of the active ingredient (Oxytetracycline hydrochloride) in all preparations was same as mentioned on the label except Oxytetracycline (74%), Terrasym PVP-100 (81%), and Onyx-LA (72%).
MIC of Oxytetracycline hydrochloride against following bacterial isolates determined by micro-broth dilution test was Bacillus subtilis (50µg), Staphylococcus aureus (100µg), Eschericiha coli (50µg), Salmonella enterica (1000µg) and Pasturella multocida (50µg).It showed that all these bacterial cultures have developed resistance against Oxytetracycline hydrochloride.
Availability: Items available for loan: UVAS Library [Call number: 1468,T] (1).
5.
Suitability Of In-House Developed Pt-Pcr Fro The Detection And Serotyping Of Dengue Virus In Pakistan
by Kashif Iqbal Sahibzada | Dr. Abu Saeed Hashmi | Dr. Aftab | Ms. Asma Waris.
Material type: ; Format:
print
Publisher: 2013Dissertation 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.
Availability: Items available for loan: UVAS Library [Call number: 1551,T] (1).
6.
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:
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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).
7.
Extraction, Purificaton And Characterization Of Proteolytic Enzyme From Fig (Ficus Carica)/ Karachi
by Haseeb Akram Sindhu | Dr. Abu Saeed Hashmi | Dr. Aftab | Ms. Faiza Masood.
Material type: ; Format:
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not fiction
Publisher: 2013Dissertation 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).
Outcomes
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.
Statistical analysis
Student's t-Test was used for comparing the means of two samples Kachri (Cucumis trigonus) and Ficus (Ficus carica).
Availability: Items available for loan: UVAS Library [Call number: 1722,T] (1).
8.
Detection Of Bacterial Load In Quail Meat Available In Lahore Market
by Muhammad Rameez akram | Dr. Naureen naeem | Ms. farasat rizwan | Prof. Dr. Aftab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2078,T] (1).
9.
Determination Of Microbial Contaminants Of Canned Fruit Products Available In Local Markets Of Lahore Pakistan
by Muhammad Waseem Akram | Dr. Muhammad Nasir | Dr. Zubair Farooq | Prof. Dr. Aftab.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2143,T] (1).
10.
Status Of Brucellosis And Its Effect On Hemogram And Serum Biochemistry In Indigenous, Cross-Bred And Exotic Dairy Cattle Herds
by Muhammad Hareem Afzal (2008-VA-250) | Dr. Muhammad Avais | Dr. Jawaria Ali Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Brucellosis mainly infects food animals such as cattle, buffalo, goats and sheep. Brucella abortus is the principal cause of brucellosis in cattle and is shed from the infected animal at or around the time of calving or abortion. The present study was conducted on 450 animals on three different strains/breeds of cattle i.e. Exotic (150), Cross-bred (150) and local cattle (150) from 10 different privately owned livestock farms of varying holdings of district Lahore. An epidemiological questionnaire focusing on herd traits as well as husbandry and sanitary practices that could be associated with the risk of Brucellosis infection was completed. Serum samples were collected and analyzed using Rose Bengal Plate Test (RBPT). The serum samples positive for Brucellosis through RBPT further subjected to Serum Agglutination Test (SAT). To check the effect of Brucellosis on hemogram, blood samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis were collected and processed for TLC, DLC, RBC, Hb, MCV, MCHC MCH and platelets using automated haematology analysed at UDL, UVAS, Lahore. Similarly, to see the effect of Brucellosis on Serum biochemistry, serum samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis collected and analysed for glucose, total protein, albumin, Creatinine, Alanine Aminotransferase (ALT), Aspartate Aminotranferase (AST) and Sorbitol Dehydrogenase (SD) using commercially available kits.
Summary
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RBPT revealed overall prevalence 17.7% higher than SAT 10.6%. Prevalence of brucellosis is higher in Cross-Bred (22.7%) followed by local cattle (18.9%) and exotic (12%).
Hemato-boichemical results showed that increase in TLC, MCV While slight changes in Hb, MCHC, RBC and values of MCV stays within normal range. On the other hand serum biochemistry increase in AST while decrease in ALT and SD found. Availability: Items available for loan: UVAS Library [Call number: 2348-T] (1).
11.
Mutational Screening Of The RB1 Gene In Pakistani Patients With Retinoblastoma
by Saeeda Kalsoom (2007-VA-555) | Dr. Muhammad Wasim) | Dr. Khushnooda Ramzan | Dr. Ali Raza Awan | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Retinoblastoma is a neonatal intraocular tumor caused by biallelic inactivation of RB1 gene. Rb
patients and asymptomatic carriers undergo a series of clinical tests for diagnosis and tumor
treatment. These clinical examinations prove to be expensive and time consuming. On the other
hand if the proband’s RB1 gene mutation status is determined by genetic testing, it can prove as
more significant and cost effective diagnostic methods. Secondly, only those asymptomatic or at
risk carriers with the mutation, require clinical surveillance while those proven to be unaffected
do not require additional clinical examinations. Furthermore early diagnosis of Rb by molecular
testing can enable and enhance clinical management, earlier treatment, follow-up care, carrier
screening, genetic counseling, prenatal diagnosis and reproductive planning in predisposed
families. Irrespective of the importance of molecular testing of Rb patients, in Pakistan only a
few clinical reports on Rb are available so, there was a dire need to find RB1 mutations in
Pakistani Rb patients and to set a molecular based diagnosis for poor affected families. Keeping
in view the importance of molecular diagnosis, in this study a reliable genetic test has been
developed to detect the RB1 germline mutations in Pakistani Rb patients.
During this study, 70 Rb patients including 38 unilateral and 32 bilateral cases were enrolled,
from different regions of Pakistan. By using direct sequencing method, seven novel and twelve
reported RBI mutations were found. The novel mutations included three frameshift mutations
(c.1116_1119delCACT in exon 11, c.1436_1437delAC in exon 16 and c.2060_2061insTCATT
in exon 20) and four substitutions (c.148G>T in exon 2, c.610G>T in exon 2, g.94G>C in exon
7, c.947A>T in exon 10 and g.1991G>C in promoter region) while twelve reported mutations in
146
22 patients included, 9 substitutions (c.160G>T in exon 2, c.289G>T in exon 3, c.751C>T in
exon 8, c.920C>T in exon 9, c.967G>T in exon 10, c.1072C>T in exon 11, c.1654C>T in exon
17, c.2063T>C in exon 20 and c.2359C>T in exon 23), one frameshift mutation (c.772_776del in
exon 8) and two splice site mutations (c.380+1G>T and c.1215+1G>A in intron 3 and 12
respectively). Mutation detection rate was found to be 77.8% in (7/9) bilateral familial, 50% in
(2/4) unilateral familial, 56.5% in (13/23) bilateral sporadic and 14.7% in (5/34) unilateral
sporadic patients while overall rate of mutations in bilateral and unilateral patients was detected
as 62.5% (20/32) and 18.4% (7/38) respectively. Beside mutations one novel c.940-64C>T
(intron 9) and nine reported intronic variants c.380+45 C>T (intron 3), c.501-77G>A (intron 4),
c.1128-72T>G (intron 11), c.1695+99A>T (intron 17), c.1695-1696delAA (intron 17), c.1815-
104A>G (intron 18), c.1961-10T>C (intron 19), c.2663+33T>C (intron 25) and c.2664-10T>A
(intron 25) were also found. Carrier screening facility was also provided to six asymptomatic
siblings (as possible carriers) of familial proband but none of them was found to be diseased.
Hopefully, in future the findings and developed protocol of this study will help to reveal the
molecular basis of Rb in Pakistani Rb patients which additionally help to secure vision and life
of Rb patients. Further, in Pakistan there is dire need to develop “National Rb Registry Centre”,
to register all new Rb cases for finding incidence rate and prevalence of Rb in Pakistan. Beside
this other related issues like financial constraints, health education, planning and awareness
about Rb, occupational training for health providers, capacity building for neonatal
ophthalmologic screening and cosmetic rehabilitation for surviving Rb patients are important and
should consider. Availability: Items available for loan: UVAS Library [Call number: 2370-T] (1).
12.
Proteomic And Genomic Analysis Of Methicillin-Resistant Staphylococcus Aureus And Efficacy Of Indigenous Medicinal Plants Essential Oils
by Sarwat Ali Raja | Prof. Dr. Muhammad Ashraf | Dr. Tayyaba Ijaz | Dr. Aqeel Javeed | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: A Cohort study (prospective and observational) was performed to study the prevalence of Methicillin resistant Staphylococcus aureus from the healthy individuals of community, hospitalized patients and associated health-care workers and indigenous plants essential oils were screened as new, improved & potent antibacterial/s against resistant strains of MRSA.
The method involved isolation and identification of MRSA from surgical wounds of hospitalized patients & associated health care workers in a tertiary care hospital in Lahore and healthy volunteers from the community. Plant essentials oils & extracts were evaluated for their antibacterial activity against selected MRSA isolates. Oils were recovered by steam distillation using an all-glass distillation assembly. Then in vitro sensitivity and MICs of plant essential oils were determined using vancomycin and linezolid as commercial standards. The essential oils were screened further for the active constituents by column chromatography using various solvents and identification of compounds were performed by GC/MS analysis and the fractions which showed prompt results were evaluated for antimicrobial activity against the MRSA isolates in quest to find new therapeutic options. Finally effective essential oils and their active fractions were studied for their toxicity using in vitro Genotoxic assays such as Ames and Comet assays. To further ensure their beneficial effects antimutagenic effect of the essential oils were also studied.
Prevalence of S. aureus among patients was 52.9%, in HCWs 86.5% and in community 74% with an overall percentage of 72.6%. Among S. aureus those declared as MRSA were 91.8% from patients, 50.6% from HCWs and 59.5% from community with an overall percentage of 62.2% MRSA. Among the isolated MRSA overall 90.6% were Coagulase positive and 75.2% were biofilm positive.
SUMMARY
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The pattern of MRSA resistance against current antibiotics have shown an overall increase in the resistance with maximum shown for lincomycin followed by tetracycline, ampicillin, fusidic acid, amoxicillin and piperacillin with tazobactam. The most effective options among current regime were tigecyclin, amikacin and meropenem showing an overall least resistance. Resistance against linezolid was observed with an overall percentage of 25.6 % and vancomycin 33.3% by disc diffusion method.
The MRSA isolates resistant to one or more groups of antibiotics were declared as MDRs. Among patients and health-care workers all were declared as MDRs where as in community 93.1% isolates were MDRs.
Upon Protein profiling using whole cell proteins 44 bands of the polypeptides were produced with molecular size 10-200kDa from the three sampling groups and were categorized into 5 clusters showing an overall significance correlation with each other explaining an interesting fact that all these strains were interlinked establishing the fact of flow of hospital acquired MRSA in the community and vice versa. This analysis also gave an insight in explaining the fact of horizontal transmission of infection within the hospital.
Keeping in view the raise in resistance among current available antibiotics indigenous medicinal plants essential oils were screened for active constituents exhibiting anti-bacterial effects against MRSA isolates.
Maximum yield was obtained from Carum copticum followed by Cuminum cyminum and minimum yield was obtained in case of Zingiber officinale. Upon qualitative analysis of all five essential oils Carum copticum essential oil showed zones of inhibition greater than the standards vancomycin and linezolid followed Cuminum cyminum and Zingiber officinale in all three
SUMMARY
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sampling groups. Anethum sowa and Myristica fragrans essential oils showed no activity against MRSA.
Minimum inhibitory concentration of the three essential oils determined by micro broth dilution method indicated that Carum copticum showed least value in all three types of MRSA isolates followed by Zingiber officinale and Cuminum cyminum.
Effective essential oils were further fractioned using silica gel gravity columns. All the fractions obtained were screened for the anti-bacterial activity against all three types of MRSA isolates. Only fraction F1 of Carum copticum showed activity greater than pure essential oil and the two commercial standards of vancomycin and linezolid.
For the identification of active constituents GC/MS analysis was performed on all three essential oils and their respective fractions. In case of fraction F1 the most dominant constituents were Carvacrol, p-Cymene, Ʈ-Terpinene and Apiol. In other two plants none of the fractions were effective. Therefore it was concluded to use pure essential oils in case of Zingiber officinale and Cuminum cyminum rather than their individual fractions and incase of Carum copticum Fraction F1 has shown superior activity.
Finally these essential oils were tested for possible mutagenic effect using bacterial reversion mutation assay and Comet assay. No mutagenic effects were observed at MIC and above doses. These effective essential oils were also evaluated for possible antimutagenic effect. Both Carum copticum and Zingiber officinale essential oils showed strong antimutagenic effects and weak antimutagenic effect by Cuminum cyminum. Upon analysis of nuclear damage none of the plants essential oils and fraction F1 of Carum copticum showed genotoxic effects and indicated to be safe. Thus from the study it was concluded that Carum copticum essential oil and its fraction F1 were the most effective to be further investigated as an alternative treatment for MRSA infections. Availability: Items available for loan: UVAS Library [Call number: 2410-T] (1).
13.
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).
14.
Pathogenesis Of Aflatoxin B1 In Quails Under Experimental Conditions And Detoxification By Biological And Chemical Means
by Sakhra Mahmood (2005-VA-251) | Prof. Dr. Muhammad Younus Rana | Prof. Dr. Asim Aslam | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Secondary metabolites of certain fungi produce toxins under favorable conditions especially while growing on different food grains. Mycotoxins are among major threats to growing poultry industry and human beings. Aflatoxins are closely related, biologically active fungal metabolites and commonly produced by Aspergillus species.
A research was carried out to evaluate the ability of Aspergillus flavus for Aflatoxin B1 production using rice, wheat and maize as substrates. Lethal effects on growth performance parameters, hematological and histopathological of graded doses of aflatoxin B1 in quails under experimental conditions were observed. Effect of Aflatoxin B1 on humoral immune response to Newcastle Disease virus vaccine in quails were determined. Biological detoxification of Aflatoxin B1 by Saccharomyces servisiae was evaluated in quails. Comparative evaluations of different commercially available toxin binders were checked. All these experiments were carried out till the six weeks (42 days).
Aspergillus flavus was identified on the basis of macroscopic and microscopic characteristics. Rice, wheat and maize grains was used as substrate to check the level of Aflatoxin B1 produced by inoculating an aqueous suspension of 106 spores/ml. Aflatoxin B1 checked by Thin Layer Chromatography (TLC) and quantified by High Performance Liquid Chromatography (HPLC).
Quails were reared under standard management conditions in five groups (A, B, C, D and E) having sixty each. Each group was further divided in two independent units. Diets offered to groups were control (without toxins), 0.25, 0.50, 1 and 2 mg Aflatoxin B1/kg feed. One unit of
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each group was vaccinated with Newcastle Disease Virus (NDV) vaccine while other was not and studied the lethal effects on growth performance, blood parameters, immune response and histopathology of vital organs. At the end of the experiment, it was found that the deleterious effects of Aflatoxin B1 were dose and duration dependent. As the level of the toxin was increased, the lethal effects were prominent. The growth performance parameters including gain in body weight, feed intake and feed conversion ratio was adversely affected at high doses. The body weight gain was significantly reduced in Aflatoxin B1 treated groups as compared to control group. Similarly feed intake and feed conversion ratio were significantly different from the control group. The hematological studies exhibited that aflatoxin B1 significantly reduced the hemoglobin, packed cell volume and total leukocyte count whereas the erythrocyte sedimentation rate was significantly increased as compared to control group. The immune response against NDV vaccine was adversely effected in Aflatoxin B1 treated groups and values of Antibody titer in AFB1 were significantly low as compared to group A( control) In the second experiment, Saccharomyces cervisae (SC) dried powder was mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. SC was added at levels of 0.5 gm, 1.0 gm and 2.0 gm /kg of feed. It was recorded that Saccharomyces cervisae (yeast) have the potential to remove the deleterious effects of Aflatoxin B1. Yeast effectively detoxified the Aflatoxin B1. The results recorded of growth performance and other parameters were non-significantly different from the control group. Chemical detoxification of Aflatoxin B1 was evaluated in quails using commercially available toxin binders. Toxin binders used were activated charcoal, kaoline, Myco AD and selenium plus vitamin E and mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. The Myco AD and selenium plus vitamin E showed the highest detoxification potential as compared
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to other chemical toxin binders. Groups E and F showed the results of growth performance, hematological, immune response and histopathological were non-significantly different from the control group (A). Kaolin was moderately detoxifying the toxin.
Presence of aflatoxin B1 in soft tissues was checked by TLC and quantified using HPLC. The liver exhibited the residues of Aflatoxin B1 at high doses of toxin. Group D and E rearing on feeds having 1mg AFB1 /Kg feed and 2mg AFB1 /Kg feed of toxin showed the residues of AFB1 in liver and kidney.
Statistical means for growth performance parameters, hematological, immune response and histopathological scores in each subunit of quails were analyzed by applying one way ANOVA and Duncans‟s Multiple Range (DMR) test at 95% probability. Aflatoxin B1 is lethal and lowers the performance of birds. The lethal effects can be detoxified by biological and chemical means to lower the economic losses to poultry industry. It can be concluded that biological detoxification is preferably better as compared to chemical detoxification. Availability: Items available for loan: UVAS Library [Call number: 2670-T] (1).
15.
Quality Assessment Of Pasteurized Milk In Relation To Time Of Different Brands Available In Lahore Market
by Noman Ali Khan (2008-VA-367) | Dr. Muhammad Nasir | Mr. Zubair Farooq | Prof. Dr. Aftab Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: There is consistent threat from adulteration, microorganism and aflatoxin M1 occurrence in the milk. Coliform, Salmonellaare among the majorfood borne pathogens which causes multiple outbreaks. This study is designed to assess the quality of different brands of Pasteurized milk sold in Lahore Market.
The quality of different brands of Pasteurized milk changes in relation to time.
The present study was conducted in Lahore market. A total of 90 samples (30 samples each day) for each of the six pasteurized milk brands were collected on 1st day and analysed on 3rd, 5th and 7th day of pasteurization. These analyses were repeated two times on monthly basis. Analytical parameters for evaluation were: physical (pH, colour, taste and odor), chemical parameters (total solid, fat, SNF and protein), adulterants (salt (NaCl), QAC, sorbitol, boric acid, hypochlorites, formalin, sugar, urea, starch, carbonates, hydrogen peroxide and detergent), microbiological (total viable count, Coliform count and detection of Salmonella) and aflatoxin M1
All the data of experimental results were analysed through repeated measure ANOVA The means will be separated with Duncan multiple range test and the level of significance will be at α=0.005 .
Availability: Items available for loan: UVAS Library [Call number: 2732-T] (1).
16.
Physico-Chemical Factors Affecting In Vitro Stability And Activity Of Phytase From Indigenous Isolate Of Asperillus Therreus
by Safina kouser (2011-VA-422) | Dr. Aftab ahmad anjum | Dr.jawad nazir | Dr.Muhammad Yasir zahoor.
Material type: Publisher: 2017Dissertation note: Phytase is commercially important enzyme. Phytate in food and feed makes it less nutritive as well as acts as anti-nutritional agent. Phytate make complexes with important mineral ions and proteins. Monogastric animals and human are not able to degrade the phytate from plant based food because they lack phytase. This leads to phosphorous deficiency. Addition of phytase into food and feed degrades the phytate. It makes, phosphorous and mineral ions become available for growth and development. There is need to evaluate these factors in vitro which in real affect the stability and activity of enzyme under feed production process and digestive system of monogastric animals.
Indigenous Aspergillus terreus isolate produce stable phytase to be used in poultry feed.Indigenous strains of Aspergillus terreus were identified by macroscopic and microscopic characteristics. These isolates were screened on Phytate Screening Medium (PSM) for phytase production. Phytase producing A. terreus was than analyzed for toxin production through TLC (Thin layer chromatography). Non toxigenic phytase producing A. terreus isolates were inoculated in phytate broth for phytase production through submerged fermentation (SmF) under optimum conditions (28°C for 8-10 days). After centrifugation and filtration supernatants were used as crude enzyme. Phytase enzyme was qualitatively analyzed through phytase assay. Phytases activity units observed for isolate PAST-16 was highest (271.49±8.14 FTU/mL) and lowest (79.00±8.05FTU/mL) of PAST-05. A. terreus phytase (PAST-16) was subjected to temperature, pH and metal ions treatment.
Thermostability of phytases was recorted at 35°C, 55°C, 75 °C and 90°C for 15, 30, 45, and 60 minutes treatments. Enzyme from A. terreus (PAST-16) was observed as thermostable at
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35°C, 55°C, 75 °C but not much stable at 95°C. Phytases showed 87.23±6.59, 198.34±4.47, 188.59±8.77 and 259.25±0.84 FTU/mL decreased in activity after 60 minutes of treatment at 35°C, 55°C, 75 °C and 95°C temperatures, respectively.
pH stability of phytases was found at pH of 2, 4, 6 and 8 for 15, 30, 45, and 60 minutes treatments. Enzyme from A. terreus (PAST-16) was observed as pH stable at 4, 6 and 8 but not much stable at 2 pH. Phytases showed 206.14±6.37, 169.59±6.37, 110.13±6.75 and 171.54±3.04 FTU/mL decreased in activity after 60 minutes of pH treatments at 2, 4, 6 and 8, respectively.
Metal ions effect on phytase activity was found with Ba2+, Ca2+, Cu2+, Fe3+, K+, Mg2+, Mn2+ and Na+ at the concentration of 1, 5 and 10mM. Enzyme from A. terreus (PAST-16) was observed as shows activity more with K+ less with Na+. Phytases showed 45.32±28.54 and 219.30±11.04 FTU/mL decreased in activity after 1mM conc. of K+ and 10mM conc of Na+, respectively.
Conclusion:
A.terreus isolate (PAST-16) produce stable phytase enzyme used in feed of poultry. In this way it tolerates condition under which feed process at commercial level and under digestive system monogastric animals. Availability: Items available for loan: UVAS Library [Call number: 2825-T] (1).
17.
Evaluation Of Antimicrobial Activity Of Essential Oil And Extracts Of Nigella Sativa Against Antibiotic Resistant Salmonella Enterica Isolates Of Human And Poultry Origin
by Sadia ashraf(2011-VA-402) | Prof. Dr. Aftab Ahmad Anjum | Dr. Ali Ahmad Sheikh | Dr.Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: The research was designed to evaluate the efficacy of methanolic and aqueous extracts of Nigella sativa, Black seed oil and thymoquinone against antibiotic resistant molecular characterized Salmonella enterica isolates of human and poultry origin (n=5 each). The compounds that have shown the antibacterial activity was also checked for their cytotoxicity by MTT assay.
Salmonella is causative agent of invasive diseases in poultry and humans, results in high mortality. Salmonellosis is a disease caused by Salmonella enterica with serious health issues related to food borne illness and most of world’s population is suffering from it. Mostly infections are treated by antibiotics but now a day’s resistance developed by Salmonella enterica. So it is need of time to develop some alternate ways to combat the problem caused by resistant bacterial pathogens. Use of essential oils and extracts of seeds are good weapons against resistant bacteria.
Salmonella enterica isolates of human and poultry origin (n=5 each) were taken from Department of microbiology UVAS Lahore and identified by colony morphology, microscopic characters, biochemical testing (Indole production test, Methyl red test, Voges Proskaeur test, Citrate utilization test and Urea utilization test) and polymerase chain reaction (PCR). For PCR product 1.5% agarose gel was run by gel electrophoresis. The biochemically identified and molecular characterized S. enterica isolates were screened for antibiotic susceptibility by Kirby Bauer disc diffusion method against amoxicillin, ampicillin, cefixime, ceftriaxone, ciprofloxacin, gentamicin, nalidixic acid, co-trimoxazole, ofloxacin and tetracycline and resistant pattern was 100% against ampicillin and Nalidixic acid and isolates shown 60% resistant against co-trimoxazole, amoxicillin and tetracycline, 80% and 40% resistant found against ofloxacin and ciprofloxacin while all isolates sensitive to cefixime and ceftriaxone. Aqueous and methanol were
CHAPTER 6
SUMMARY
used as solvents for extraction from Nigella Sativa. Seeds were dried, mixed, centrifuged, filtered and filtrate evaporated to obtained extracts. Percentage yield of methanolic extract was more than aqueous extract. Commercially available black seed oil, thymoquinone, water and methanol extracts of black seed would be evaluated for antibacterial activity by well diffusion method. Zones were measured in millimeters. All compounds gave the zones of inhibition except aqueous extract against Salmonella enterica isolates. The minimum inhibitory concentration (MIC) was determined by micro broth dilution method and then methanolic extract, black seed oil and thymoquinone used in MTT assay to evaluate their cytotoxicity. Cell survival percentage was calculated by a formula.
Data were analyzed by one way analysis of variance (ANOVA) followed by Duncan’s multiple range tests (DMRT) using statistical package for social sciences (SPSS version 20.0). Antimicrobial activity of essential oils, thymoquinone, water and methanol extract would be compared by graph pad prism 5.0 statistical software. Availability: Items available for loan: UVAS Library [Call number: 2827-T] (1).
18.
Determination Of Tartrazine In Different Food Items Available In Lahore Pakistan
by Anam Arshad (2015-VA-1055) | Dr Zubair Farooq | Dr. Sanaullah Iqbal | Prof. Dr. Aftab Ahmad Anjum.
Material type: Publisher: 2017Dissertation note: Synthetic dyes used in various food items like sweets, candies and beverages cause severe health hazards if they are not food grade and within the legally permitted limits. In Pakistan due to increased consumption of attractive colored food items, the use of dyes in every food product is also increasing. Already there is no study to appraise the nature and level of colorants. Therefore, this study focused on determination of synthetic dye (tartrazine) used in candies, sweets and beverages to determine its safer levels.
This research work was done in food science and nutrition lab of Food Science and Human Nutrition Department of UVAS, Lahore, Pakistan. Total 180 samples were collected from all 9 towns of Lahore plus Lahore Cantt. Samples included 30 candies purchased from local vendors and 30 candies from shops, 30 sweet samples (rasgulla) from local vendors and 30 sweet samples from sweet shops. Moreover, 30 slush samples locally available in streets and 30 slush samples from shops of posh areas in Lahore. The results were compared with the previous held researches in other countries.
The food samples were divided into two categories local shops and local vendors. Total six local shops and six vendors of each town of lahore were selected for the collection of samples in triplicate pattern. All the samples were evaluated for spectrophotometric determination of tartrazine by using Beer’s law. Abosrbtion peeks were checked at a wavelength of 421.6 and the mean values of concentration of tartrazine in slush ranged from 0.269 to 0.275 mg/g obtained from local vendors and shops respectively.Moreover, the mean values of tartrazine ranged from 0.258 to 0.309 mg/g for vendor sweet and shop sweet samples respectively and mean value for candies ranged from 0.200 -0.704mg/g. Data was analyzed through the Microsoft Excel 2010 and SPSS 22.0. Mean values with standard deviation in percentages of results were analyzed by descriptive analyses. To examine the relationship among the variables of candies, sweets and slush samples chi-square test was used. Further to compare tartrazine levels in the local shops and foodstuff obtained from the common vendors of Lahore, independent “t” test was used. 2 way-ANOVA was applied to check the differences of all samples in 10 towns of Lahore.
According to my investigations the quality of foodstuff collected from local shops and from local vendors is almost same and both contain high amounts of tartrazine.I suggest consumers, they should prefer to buy branded foodstuff from superstores as compared to local shops and local vendors because the keeping quality and handling practices are good in superstores than local shops.
Availability: Items available for loan: UVAS Library [Call number: 2886-T] (1).
