1.
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:
print
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).
2.
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).
3.
Status Of Brucellosis And Its Effect On Hemogram And Serum Biochemistry In Indigenous, Cross-Bred And Exotic Dairy Cattle Herds
by Muhammad Hareem Afzal (2008-VA-250) | Dr. Muhammad Avais | Dr. Jawaria Ali Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Brucellosis mainly infects food animals such as cattle, buffalo, goats and sheep. Brucella abortus is the principal cause of brucellosis in cattle and is shed from the infected animal at or around the time of calving or abortion. The present study was conducted on 450 animals on three different strains/breeds of cattle i.e. Exotic (150), Cross-bred (150) and local cattle (150) from 10 different privately owned livestock farms of varying holdings of district Lahore. An epidemiological questionnaire focusing on herd traits as well as husbandry and sanitary practices that could be associated with the risk of Brucellosis infection was completed. Serum samples were collected and analyzed using Rose Bengal Plate Test (RBPT). The serum samples positive for Brucellosis through RBPT further subjected to Serum Agglutination Test (SAT). To check the effect of Brucellosis on hemogram, blood samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis were collected and processed for TLC, DLC, RBC, Hb, MCV, MCHC MCH and platelets using automated haematology analysed at UDL, UVAS, Lahore. Similarly, to see the effect of Brucellosis on Serum biochemistry, serum samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis collected and analysed for glucose, total protein, albumin, Creatinine, Alanine Aminotransferase (ALT), Aspartate Aminotranferase (AST) and Sorbitol Dehydrogenase (SD) using commercially available kits.
Summary
62
RBPT revealed overall prevalence 17.7% higher than SAT 10.6%. Prevalence of brucellosis is higher in Cross-Bred (22.7%) followed by local cattle (18.9%) and exotic (12%).
Hemato-boichemical results showed that increase in TLC, MCV While slight changes in Hb, MCHC, RBC and values of MCV stays within normal range. On the other hand serum biochemistry increase in AST while decrease in ALT and SD found. Availability: Items available for loan: UVAS Library [Call number: 2348-T] (1).
4.
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).
5.
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
Summary
74
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).
6.
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).
7.
Acaricide Resistance Of Tick Population Infesting Buffaloes In District Narowal
by Muhammad Mubashir Abdullah (2015-VA-1104) | Prof. Dr. Kamran Ashraf | Dr. Muhammad Latif | Prof. Dr. Aftab Ahmad Anjum.
Material type: Publisher: 2017Dissertation note:
Tick imperviousness to acaricides is an expanding issue in Pakistan and represents a genuine financial danger to the domesticated animals and veterinary pharmaceutical enterprises. New acaricides are to a great degree costly to grow so the present acaricides ought to be viewed as a constantly decreasing asset, which ought to be ensured by all methods conceivable. The principle goal of the review was to distinguish the stages of tick imperviousness to acaricides at close business and collective ranges in District Narowal, Pakistan. Likewise to contrast the in vivo techniques and with explore acaricide administration procedures which may build the life expectancy by utilized acaricides.
To meet these points a field survey (February 2016 to March 2017) was carried out at 3 tehsils (Tehsil Narowal, Shakargharand Zafarwal cities of Pakistan to monitor levels of field tick resistance to acaricides. The larvae were originally obtained from engorged female A.hebraeum, Hyalloma, Boophilus, Dermacentor, Ixodes, R. appendiculatusand R. evertsievertsi. The larvaewere tested against different concentrations of trichlorofon, ivermectin and cypermethrin using the Shaw Larval Immersion Test (SLIT). Mortality dose data were subjected to probit analysis using a BMDP statistical package. Factors of resistance (FOR) were calculated by comparing the larval response of ticks from the field.
On the communal farms high levels of tick resistance were detected to cypermethrin as well as partial resistance to ivermectin whilst no resistance was detected against trichlorofon. On the commercial farms, however, ticks were equally resistant to trichlorofon, cypermethrin and ivermectin. The populations of Hyalloma, Boophilus, Dermacentor, Ixodes, on these farms had developed higher levels of resistance to the testacaricides than the equivalent R. evertsievertsi, R. appendiculatus and A.hebraeumpopulations. Higher levels of tick resistance to trichlorofonwas observed on3 tehsils (Tehsil Narowal, Shakarghar and Zafarwal)than on communal farms, however, there was no significant differences in tick resistance to ivermectin and cypermethrin at both the commercial and communal farms. It was surmised that inappropriate use of acaricides might have resulted in higher tick resistance to the currently available acaricides on the commercial as well as the communal farms. Correct acaricide usage may solve this problem to a limited extent.
Comparative in vivo tests were also carried out on the larvae and adults of Hyalloma, Boophilus, Dermacentor, Ixodes, to determine the susceptibility of this tick to different concentrationsof the currently used acaricides, (amitraz, ivermectin and cypermethrin) at three commercial dairy farms, (“Brycedale”, “Sunny Grove” and “Welgevind”) in the areas of District Narowal, Pakistan. Resistance of field strains of Hyalloma, Boophilus, Dermacentor, Ixodes, Dermacentor,were determined using the Adult Immersion Test (AIT) as the latter test took into account factors such as oviposition assessment and reproductive ability. At “Brycedale”, resistance to trichlorofon and ivermectin was detected with the AIT method. Emerging resistance to trichlorofon and resistance to ivermectin were also detected . At “Sunny Grove” resistance was detected to cypermethrin and at “Welgevind” resistance was detected to ivermectin with the SLIT whilst no resistance was detected using AIT. It would appear that the Hyalloma, Boophilus, Dermacentor, Ixodes, populations tested on these dairy farms were more resistant toivermectin than to trichlorofon or cypermethrin.
Nearly 50% of the dairy farms sampled showed resistance to ivermectin and the majority had susceptible Hyalloma, Boophilus, Dermacentor, Ixodes, populations to both amitraz and cypermethrin. In general there was a good correlation between the Cypermethrin and Trichlorofon whilst in many cases there was poor correlation between the Cypermethrin and Ivermectin.
From this study it would appear that the In vivo method was a reliable to detect resistance within seven days. In vitro method the ELT and the RET could possibly be used as screening methods to detect acaricide resistance on farms whilst the SLIT would remain the test of choice for National surveys. In addition the ELT is less costly and does not require sophisticated equipment for field testing if resistance development compared with other in vitro test methods. This method, however, still needs to be validated and standardized for use in Narowaland the rest of punjab where tick control is important.
Availability: No items available
8.
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).
