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1. In-Vetro And In-Vivo Anti-Theilerial Activity Of Medicianal Plants

by Mukhtar Ahmad | Prof. Dr. Muhammad Ashraf | Prof. Dr. Muhammad Sarwar Khan.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2013Dissertation note: In vitro study was carried out in order to estimate the anti-theileria effect of Calotropis procera and Peganum harmala. Water and chloroform extracts of each plant were used in-vitro study along with standard drug Buparvaqoune (Butalex®). For this four concentrations i.e.4, 8, 12 and 16mg/ tested solutions of each extracts of each plant were applied on cultured lymphocytes exposed to theileria parasite infection. ELISA reader findings showed that the mean OD were found less in treated theileria infected lymphocytes cell culture as compared to untreated culture wells mean OD values. The highest cell reduction (94.36%) with C. procera chloroform extract treatment was observed at a concentration of 16 mg/ml solution. Lowest concentration (1mg/ml solution) of C. procera chloroform extract reduced non-significant (P>0.05) lymphocytes cell proliferation (40.97%) as compare to control negative group. P. harmala water extract was effective against the theileria parasite as significantly lower (P<0.05) mean OD value (1.802 ±0.341) was measured at a concentration of 4mg/ml solution and maximum inhibitory effect (92.20) was seen at a concentration of 16 mg/ml solution. ELISA reader findings showed that P. harmala chloroform extract treatment failed to inhibit lymphocytes cells propagation even at highest concentration. The highest inhibitory effect (85.33%) against theileria infected lymphocytes propagation was seen at a concentration of 16 mg/ml solution. Plant extract was evaluated in respect of feed intake in rabbits. It showed that when administered extracts of C. procera in rabbits at dose of 3 mg and 5 mg/kg body weight, did not affect on feed intake in rabbits. However the chloroform and water extracts of both plant i.e. C. procera and P. harmala when were administered in rabbits parentally at dose 10 of mg/kg body. It showed that the feed intake of rabbits was non-significantly reduced as compared to other treatments groups. Hematological parameters such as WBC X103 count, RBC X 106 count and Hb g/dl values were measured at various days. Findings showed that significantly lesser RBC X 106 count was in group A3 and D3 than control at day 30 of experiment. A non-significant difference (P>0.05) was seen in RBC X 106 count and Hb g/dl measurements in all treatments groups. Kidney and liver functions were evaluated by measuring biochemical parameters, uric acid, creatinine and ALT at 0 days, 9 days and 30 days. Findings showed that serum creatinine and urea enzyme levels were significantly higher (P<0.05) in group A3 as compare to control group at day 30 of experiment. Serum level of urea was also significantly higher (P<0.05) in group B3 and D3 at day 30 of experiment. A non-significant difference (P>0.05) was seen in ALT in all treatment groups at day 30 than control. Post-mortem was performed at day 30 of experiment. Gross lesions consisting of hemorrhages, congestion, and lung emphysema were seen in rabbits treated with high dose i.e. 10 mg/kg of both extract of C. procera. Rabbits treated with P. harmala chloroform extract at dose 10 mg/kg showed moderate gross lesions. Histopathology of organs such as lungs, kidney, liver and heart was performed. Toxicity lesions were seen in rabbits treated with high dose i.e. 10 mg/kg of both extract of C. procera. Rabbits treated with P. harmala chloroform extract at dose 10 mg/kg showed histopathological lesions in lungs, liver and kidney. Theileria infection was studied in vivo by developing through theileria infected Hayalomma ticks in crossbred calves (n=30) through. At day 15 of infection maximum increase in mean rectal temperature (105.24 ± 0.46F) was observed, twenty four calves had pyrexia (104.1- 105.6 F) and six claves were showing pyrexia > 105.6 F. A significant increase (P<0.05) in pre-scapular lymph node enlargement score of challenged calves was seen by day 7 of infection and maximum lymph node score (grossly enlarged size) was noticed in twenty calves (Table 4.14 , Plate 4.16) with peak mean score (2.73±0.44) on day 13 of infection. The piroplasm peak score (3.80±0.83) was observed in challenged calves at 22 day of infection and remained significantly higher (p<0.05) (2.60±0.54) in untreated calves until the 36 day of infection (Fig.4.29 and Table 4.16). A significant increase (P<0.05) in mean schizonts was observed in pre-scapular lymph node biopsy smear from day 7 of infection to onwards. Blood samples of challenged calves (n =30) were confirmed theileria positive through PCR test. The amplification of Theileria species were amplified at 1098 bp (Plate 4.20 and Theileria annulata was amplified at 721 bp (Plate 4.21). In order to estimate the pattern of disease severity, severity score was measured by summation of mean score of piroplasms, schizonts, lymph node swelling and rectal temperature. From day 7, mild response (3-5 score) was seen in infected calves (n=10). With increase in the severity of disease a significant decrease (P<0.05) was observed in mean values of the Hb g/dL amount, WBC and RBC count, Hct (%) concentration and lymphocytes percentage from day15 of infection onward to 36 day of infection. A non-significant decrease (P>0.05) in the mean values of MCH pg was seen throughout the experiment. A significant decrease (P<0.05) in mean values of MCHC g/dL along with significant increase (P<0.05) in the mean measurement of MCV fL (64.14±3.53) values was seen at day 36 of infection as compare to day 0 values, indicating macrocytic hypochromic anemia in challenged calves. These findings showed a significant increase (P<0.05) in excretory products (uric acid and creatinine) from day 15 of infection and onward as compared to day 0 values, indicating damaged kidney in infected calves. Biochemical analysis showed the significant increase (P<0.05) in liver enzymes (ALT, AST) from day 15 infection and onward. Anti-Theileria activities of drugs were estimated by evaluating clinical manifestation of the disease and parasitological findings. Beside this treatment effect on hematological and biochemical reactions of liver and kidney functions was determined. A significant difference (P<0.05) in rectal temperature of calves groups (B and E) was observed than control positive (group F) at day 21 of post-treatment. On other hand calves treated with treatments A, C and D had a non-significant difference (P>0.05) in rectal temperature compared with untreated calves (group F). It was found that calves (n=5) dosed with C. procera chloroform extract (group A) had rectal temperature in normal range by the day 7 of post-treatment. Similarly calves (n=5) treated with Butalex were found with normal rectal temperature from the day 7 of pos-treatment. On other hand, at day 21 of treatment 40%, 20%, 40% and 80% calves were found with pyrexia in treatments groups B, C, D and F, respectively (Table 4. 46). By the day 14 of treatment, calves of treatment groups B and E showed no parasitemia (piroplams ?1). Disease severity was estimated on accumulative score of rectal temperature, lymph node swelling and parasitological findings (piroplasms and schizonts score). It was found a significant decrease (P<0.05) occurred in the disease severity of score of disease in calves of groups B and E as compare to A, C and F at day 3 of post-treatment. At day 21 of treatment all treated calves were recovered from anemia. Availability: Items available for loan: UVAS Library [Call number: 1580,T] (1).

2. Influence Of Early Weaning On Growth Performance, Plasma Metabolites And Rumen Fermentation Indices In Neonatal

by Muhammad Afzal Rashid | Prof. Dr. Talat Naseer Pasha | Prof. Dr. Makhdoom Abdul Jabbar.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2013Dissertation note: Rearing of young calves is a labor intensive and costly segment of livestock production. From birth to weaning, young calf undergoes a transition from monogastric to adult ruminant. The concept of weaning from milk at an early age is based on early development of functional rumen enabling calves to utilize low quality roughages. In current study, a series of experiments were conducted to refine the more effective weaning regime for buffalo calves and use of by-products of the ethanol production industry in early weaned cattle calves. Conventionally, buffalo calves are kept with the dam, allowed to suckle a little amount of milk along with seasonal green forages, and weaned around the age of one year. To date, limited published work was available on growth performance and economics of buffalo calves weaned from milk at an early age. Therefore, the experiment was conducted to reduce the weaning age and evaluate the growth performance of male Nili-Ravi buffalo calves. Twenty-four male buffalo calves were assigned to one of the three treatments: continuous milk feeding (CMF), limited milk feeding (LMF), and early weaning (EW). After colostrum feeding, calves were individually fed whole milk at 10% of their BW, adjusted weekly until 6 wk of age. Thereafter, milk allowance was gradually tapered to zero in CMF, LMF and EW treatments at 12, 10 and 8 wk of age, respectively. Calf starter feed was provided ad libitum from wk 2 through wk 12 and individual intakes were recorded daily. Blood sampling was carried out form wk 6 through 12, on a weekly basis. The BW and structural measurements (HG, WH, and HW) were carried out at the start of experiment and later on a weekly basis. In young buffalo calves, the regimen of weaning at 8 weeks of age was more effective. The early weaned calves showed similar growth rate to those in the CMF and LMF by consuming more calf starter and saving a substantial amount of high priced milk. On the basis of the results of this experiment, buffalo calves successfully adapted to early weaning that might help to mitigate issues like poor growth and low returns associated with traditional calf rearing practices. Furthermore, this study effectively reduced the weaning age from 1 year to 8 weeks of age. Hence, reducing weaning age did not affect the growth performance of Nili-Ravi buffalo calves by 12 weeks of age. Early development of the rumen is the main objective of a successful early weaning program which depends upon the amount of starter intake, VFA production, and ruminal papillae development. Studies have shown that grains in starter feed can be replaced by DDGS up to 28% of DM without compromising the growth performance and rumen development. Second experiment was planned to evaluate the effects of replacing grains and soybean with DDGS and ammonia treated DDGS at 25% of DM. Study was conducted in collaboration with dairy science department SDSU (USA). Twenty one neonatal male Holstein calves were assigned to one of the three of dietary treatments: C = 0% DDGS, DDGS = 25% DDGS, CAFEX-DDGS = 25% CAFEX treated DDGS. In a 10 week experiment, calves were fed 680 g MR through 4 week, reduced to half during wk 5, and weaned at the end of wk 5. Starter intakes were conducted daily; whereas, body weights, structural measurements were conducted at the start of experiment and then on a weekly basis. Jugular blood samples were taken on a weekly basis using EDTA and NaFl coated evacuated tubes. Rumen samples were collected from a subset of 15 calves (n=5 calves/ treatment) at wk 5, 7 and 10. At the end of experiment, four calves from each treatment were also slaughtered to determine rumen morphometric measurements (PL, PW, RWT and PC). Experiment illustrated that weight gain, structural measurements, total starter intake, DMI and feed efficiency were not affected by the inclusion of DDGS and CAFEX treated DDGS at 25% of DM in starter feeds. CAFEX treatment of DDGS improved the CP contents of DDGS from 29.5% to 40%; however, inclusion of CAFEX-DDGS in starter reduced feed intake during the pre-weaning period. Whereas, overall starter intake was higher in calves fed DDGS based starter feed indicating the effect of ammonia treatment on palatability. Lower pre-weaning starter intake, slow rumen fermentation of CAFEX-DDGS resulted in lesser BHBA concentration leading to lesser development of rumen papillae growth (PL and PW). However, there was a tendency for higher weight gain in calves fed DDGS based starter due to increase in starter intake. In the light of these results it is concluded CAFEX-DDGS can be included in starter feeds at 25% of DM without affecting the growth performance. However, further research is required to evaluate the digestibility of DDGS after CAFEX treatement. Similar, growth performance indicates that CAFEX-DDGS can replace the corn and soybean meal in starter feeds. In third experiment, microbial diversity in developing rumen and intestine of young calves fed DDGS and CAFEX treated DDGS at 25% of starter was investigated. Experiement was carried out at SDSU dairy research station (USA). Fifteen calves with n=5 per treatment, fed according to protocols described in Experiment II. Calves were sampled for rumen contents at wk 5, 7 and 10 of age; whereas, intestinal contents were collected at the time of slaughter. The DNA was extracted subjected to PCR-DGGE and dendogram was constructed using cluster analysis software. Results revealed that microbial population was highly different from each other at wk 10 indicating the effect of age and dietary treatment on rumen micro flora. Whereas, intestinal and rumen bacterial diversity at wk 5 and 7 of age was not affected by inclusion of DDGS and CAFEX-DDGS in starter feed. The changes in intestinal microflora of DDGS and CAFEX-DDGS fed calves compared with control group showed that the effect of dietary treatments on post-ruminal availability of nutrients and microbial proteins. In conclusion, rumen bacterial population changes with the advancing age and the type of ingredients used in the diet. Further, research is required to identify the effect of feeding DDGS on growth of particular bacteria like methanogen and their impact on methane production and feed efficiency. Availability: Items available for loan: UVAS Library [Call number: 1595,T] (1).

3. Pathology Of Experimental Enterotoxemia In Sheep And Goats

by Azam Ali Nasir | Prof. Dr. M. Yonus Rana | Dr. Muhammad | Dr. Muti-ur Rehman Khan.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: During the present study intestinal scrapings were collected from sheep suspected for enterotoxaemia. Samples were subjected for the isolation by repeated culturing in Reinforced Clostridium medium. Growth on blood agar revealed characteristic colonies of Clostridium perfringens after 18-24 hours. Biochemical and mice inoculation tests were performed. Isolated organism was identified by indirect ELISA. The pure growth was subcultured in RCM in bulk quantity and calculated the dose for experimental infection. In the second part of the study, sheep and goats were procured, dewormed and kept in the experimental house of UVAS, Lahore. The experimental infection comprised of the whole culture of C. perfringens type D was inoculated intraduodenally via para-mid line between animals of group A and B while in animals of group C and D inoculated only starch solution to achieve the objectives. Accumulative clinical score in sheep was found to be 9 to 16, 13 to 22 and 15 to 23 at 10, 20 and 30 hours PI respectively while in goats the accumulative scores varied from 5 to 15, 9 to 16 and 14 to 21 at above mentioned time intervals. The highest mean score for clinical findings in sheep was anorexia, frothing followed by dehydration while in goats, the highest mean score was recorded for diarrhea dehydration, and anorexia. No significant clinical findings were noted in control groups. There was a significant increase in blood glucose, urea and serum creatinine in infected group of sheep and mean values reached up to 141 mg/dl, 92 mg/dl and 7.5 mg/dl respectively at 30 hours PI while in goats a similar pattern was observed with the mean values raised to 142 mg/dl, 111 mg/dl and 10.2 mg/dl for blood glucose, urea and serum creatinine respectively. There was no significant change found in RBC and platelet count of both species but there was an early increase in the mean WBC count of sheep 19.7x103/µl at 10 hours but then decreased to 14.7x103/µl at 30 hours PI while in goats it was 23.6x103/µl and then decreased to 15.3x103/µl. The mean PCV % age increased in animals of both infected groups but more in goats and reached to 52% in 30 hours. During the third part of the present study, the animals were slaughtered and PME performed. The accumulative score for gross lesions were recorded and it was found between 14 to 24 in animals of group A with the highest score for congestion and edema of different organs whereas in goats it was between 12 to 22 with congestion and hemorrhages of intestine having highest scores. The samples were kept in formalin for histopathological examination and accumulative lesions score was noted in different organs. The highest mean score in sheep was recorded in kidneys and lungs and in goat intestine, lung and kidneys were the major organs affected. A polymerase chain reaction was optimized under our own laboratory condition for the detection of alpha and epsilon toxins of Clostridium perfringens type D from different tissues. Alpha gene was amplified at annealing temperature 52.2oC with amplicon size 247bp and ETX gene at the annealing temperature. 50.2oC with amplicon size 665bp. The erythrocytes of different species were used to know the sensitivity against culture supernatants of C. perfringens type D. It was observed maximum hemolysis occurred in human erythrocytes (68%) followed by mice (57%) at 37oC. It was also recorded that a significant increase was found at 37oC as compared to25oC except for dog and rabbits where no significant difference was observed. Availability: Items available for loan: UVAS Library [Call number: 1596,T] (1).

4. Phenotypic And Genetic Aspects Of Some Performance Traits Of Buchi Sheep In Pakistan

by Maqsood Akhtar | Prof. Dr. Khalid Javed | Prof. Dr. Muhammad Abdullah.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1598,T] (1).

5. Seasonal Variations In The Testicular And Seminal Characteristics Of Cholistani Bulls

by Umer Farooq | Prof. Dr. Ijaz Ahmad | Dr. Hafsa Zaneb | Prof. Dr. Habib Rehman | Faculty of Biosciences.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2013Dissertation note: All the indigenous cattle of Pakistan belong to Zebu (one-humped) cattle (Bos indicus). There are 15 recognized indigenous breeds of cattle which constitute 43% of the total cattle population in the country. Hefty work has been documented on many aspects of Sahiwal and Red Sindhi breeds of cattle in their respective local climates which have rightly gained them an international fame as being the vital tropical milk breeds of Pakistan. However, scanty work has so far been reported on reproductive indices of other indigenous cattle breeds of Pakistan such as Dajal, Cholistani etc under their local climatic conditions, and their potential still remains unearthed. Hence, it is the dire need of time to comprehend the urgency of protection and propagation of the indigenous cattle breeds of Pakistan at every front. Following the global lead, an effort has been made through the present study to highlight a formerly neglected indigenous cattle breed of Pakistan- Cholistani- being reared by the nomadic herders of the Cholistan desert, Pakistan. The study was executed with a specific objective to assess baseline data, seasonal variations and correlations between various non invasive, readily measurable reproductive [age, body weight (BW) and orchidometric parameters viz. scrotal circumference (SC), scrotal skin fold thickness (SSFT), average testicular length (Avg L), average testicular width (Avg W) and paired testicular volume (PTV)], seminal (fresh semen and seminal plasma) and haematobiochemical attributes of Cholistani AI bulls (n=06). Keeping in view the prevailing climatic conditions of the study area, 4 seasons of 2 months duration each were defined as i) stress free autumn (October-November), ii) stressful winter (December-January), stressful dry summer (May-June) and iv) stressful wet summer (July-August). The overall mean values (± SEM) for age, BW, SC, SSFT and PTV in the present study were 92.83±2.63months, 527.55±4.5kg, 35.39±0.28cm, 1.03±0.02cm and 851.84±16.34cm3, respectively. The BW was recorded to be significantly lower (P<0.05) during the stress free autumn (507.08±8.26kg), however no significant difference was found in the remaining stressful seasons of winter, dry and wet summer. The SSFT was significantly higher (P<0.05) in stress free autumn (1.07±0.04cm) and winter (1.13±0.05cm) being followed by that in wet summer (0.97±0.02cm) and dry summer (0.94±0.03cm). SC and PTV were found to be constant during all the study seasons with no significant variation. Significantly positive correlations (P<0.01) were noticed between various traits. The overall mean values (± SEM) for ejaculatory volume, number of ejaculates per bull, mass motility, individual sperm motility, number of dozes frozen per bull, sperm concentration, live sperm, morphologically normal sperm and those with acrosome integrity in the present study were 4.92±0.14mL, 1.89±0.02, 2.26±0.06, 63.51±1.03%, 208.26±8.42, 989.73±27.4million/mL, 86.74±0.55%, 86.34±0.51% and 86.64±0.48%, respectively. In general, all the seminal attributes revealed significantly (P<0.05) better results in stressful seasons as compared to stress free season, with the exception of number of ejaculates per bull which was not affected significantly by seasons. In terms of semen production, a total of 39778 doses of 0.5 mL straws were frozen during the study period. Significantly higher number of doses per bull (P<0.05) was frozen during dry (245.37±17.84) and wet summer (250.37±17.75) as compared to autumn (191.4±14.22) and winter (143.92±11.25); the difference between the latter two seasons was also significant, while it was non significant between the former two seasons Regarding the subjective analysis of spermatozoa, higher (P<0.05) mean values were recorded for all three parameters (live, morphologiclly normal and acrosome intact sperm) during stressful winter and dry summer as compared to wet summer and autumn, the difference between the former two seasons was, however, non significant. Moreover, mean values for these attributes were significantly higher during wet summer as compared to autumn. Amongst the physical attributes of semen, only mass motility revealed a significantly positive (P<0.01) correlation with individual sperm motility (r 0.619). However, regarding the subjective analyses, live sperm had a positive correlation with normal (r 0.848) and with acrosome integral sperm (r 0.790). In seminal plasma chemistry, the overall mean values (± SEM) for Na+, K+, cholesterol and glucose recorded in the present study were 236.41±4.2, 142.95±3.81, 152.02±3.17 and 127.22±3.65 mg%, respectively. Amongst these, Na+ was significantly higher (P<0.05) during dry summer (270.35±8.89mg%) whereas K+ was higher in dry summer (169.1±5.54mg%) and winter (154.80±6.29mg%). Cholesterol and glucose were found to be significantly elevated (P<0.05) during dry summer season being 173.7±4.24 and 147.95±7.74mg%, respectively. Na+ had a significantly positive (P<0.05) correlation with glucose (r 0.232). Similarly, positive (P<0.01) correlations were seen for K+ with Na+ (r 0.341) and cholesterol (r 0.390). The overall mean values (± SEM) for Hb, TEC, PCV, MCV, MCH and MCHC recorded for the Cholistani AI bulls in the present study were 12.24±0.15mg/dL, 6.28±0.82×106/µL, 37.3±0.4%, 59.77±0.61fL, 19.58±0.2pg and 32.89±0.33g/dL, respectively. Regarding the white blood cell values, the overall means (± SEM) for TLC, neutrophils, lymphocytes, monocytes and eosinophils were 7.58±0.17×103/µL, 40.45±1.05%, 53.2±0.85%, 4.05±0.47% and 2.3±0.26%, respectively. DLC, in the present study, revealed that the lymphocytes were the predominant leukocytes followed by neutrophils. No significant effect (P<0.05) of season was noticed on any of the red blood cell values studied. However, TLC amongst the white blood cell values was found to be significantly higher in dry summer (8.16±0.28×103/µL). The overall mean values (± SEM) for blood serum NA+, K+, cholesterol and glucose recorded for the present study were 130.73±0.91, 7.37±0.11, 82.8±1.52 and 75.22±2.04mg%, respectively. Correlation analysis revealed significant positive correlations between mass and individual sperm motility, morphologically normal and live sperm and sperm acrosome integrity and live sperm (P<0.01). Among biochemical attributes of seminal plasma, Na+ and K+, cholesterol and K+ and glucose and Na+ were correlated (P<0.01). Only seminal plasma cholesterol was positively correlated with ejaculatory volume (P<0.01). An overall mean value of 5.81±0.32ng/mL was recorded in the present study for testosterone being non significantly effected by seasons. All its correlations with all attributes studied were found to be non significant. In conclusion, various physical and biochemical attributes of Cholistani bull semen were better in stressful seasons as compared to stress free season. The acceptable quality of semen of Cholistani bulls recorded during wet/dry summer in the present study is an indicative of the fact that this breed has an innate ability of being well adapted to the harsh, hot desert climate. Furthermore, the present study revealed that the Cholistani breeding bulls have an amazing tendency to maintain most of their haematobiochemical parameters at a near constant level without showing much variation during stress free or stressful times. Availability: Items available for loan: UVAS Library [Call number: 1739,T] (1).

6. Immunobiological And Molecular Characterization Of Pasteurella Multocida From Buffaloes

by Muhammad Kamran | Prof. Dr. Mansur-ud-Din Ahmad | Dr. Aftab Ahmad Anjum | Prof. Dr. Azhar.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: Hemorrhagic septicemia is an acute bacterial disease of buffaloes and cattle caused by Pasteurella multocida. In the present study, 400 samples (200 from carriers and 200 from sick animals) from Sargodha division were collected. Among four districts of the division, 15 samples were positive by API Kit, 13 by conventional biochemical tests and eleven were found positive for P. multocida through serological and molecular characterization. Biochemical profile index obtained with API kits had lesser accuracy than conventional and serological profiles for the identification of P. multocida. Passive mouse protection test and AGPT were used for serological confirmation. Different molecular techniques like SDS-PAGE, PCR and RFLP were used to investigate variation at the molecular level in field and vaccinal strains. There were no significant variation between field isolates and vaccinal strain in sick animals and carriers, or in isolates of different districts. Five major and three minor polypeptide bands were observed by SDS-PAGE. Genetic relatedness among the isolates was assessed by cluster analysis using Fingerprint Analysis of Missing Data (FAMD) of 12 isolates. The12 isolates clustered into 5 groups namely I, II, III, IV and V. Group I and II consisted of only one isolate in each (8.33%) of the total designated BKC-01 (S5) and KBO-01 (S1), respectively. Group III composed of 2 isolates (16.67%) namely KBC-02 (S4) and MNO-01 (S2). Group IV had the highest numbers of isolates (50%) designated as KBC-02 (S3), MNO-01 (S6), BKO-02 (S7), MNC-02 (S8), SGO-02 (S9) and V. Only two isolates were typed in group V (16.67%) named as SGO-01 (S10) and BKO-01 (S11). The size of amplified gene was 460 bp. HindIII I endonuclease cleaved bacterial genome at four sites as compared to other four enzymes (DNase1, PstlI, EcorI and BamHI) change the writing of these enzymes which cleaved at two sites. The isolates were also subjected to ten routinely used antibiotics for sensitivity testing and found enrofloxacin as drug of choice with 90.91% sensitivity, followed by gentamycine, chloramphenicol, ciprofloxacine and norfloxacine (72.73%), ampicillin and amoxycillin (45.45%), amikacin (36.36%) and lowest to sulfadiazine and erythromycine (18.18%). Availability: Items available for loan: UVAS Library [Call number: 1767,T] (1).

7. Chemical Characterizaton And Toxicological Screening Of Auto-Rickshaw Emissions Particulate

by Khaleeq Anwar | Prof. Dr. Muhammad Ashraf | Dr. Aftab | Dr. Aqeel Javeed.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2013Dissertation note: Vehicular air pollution is a mounting health issue of the modern age, particularly in urban populations of the developing nations. Auto rickshaws are not considered eco-friendly as to their inefficient engines producing large amount of particulate matter (PM), which poses a significant environmental threat. Major transformations in the environmental composition are principally attributable to the combustion of fuels by automobiles. Motorized gasoline powered two-stroke auto-rickshaws (TSA) and CNG powered four-stroke auto-rickshaws (FSA)are major sources of air pollution in south Asia and produce toxic amount of PM to the environment. In this study, during the first phase, the PM of TSA and FSA was characterized by using proton induced x-ray emission (PIXE) analysis. The observations of the existing investigation recognized significant increase in Al (P < 0.05), P (P < 0.01), and Zn (P < 0.01) from the PM samples of FSA. In addition, the concentrations of Cu, Fe, K, Mn, Mg, Na, S and Si were also observed exceeding the recommended NIES limits. On the contrary, increased concentration of Sr and V were observed in the PM samples from TSA. It is generally believed that FSA generates smaller amount of PM but the data obtained from this study clearly shows that emissions from FSA are comprised of potentially more toxic substances than TSA. The current research is specific to the metropolitan population and has evidently revealed an inconsistent burden of exposure to air pollutants engendered by FSA in urban communities, which could lead to disruption of several biological activities and may cause severe damage to entire ecological system. The second phase of this study was conducted to ascertain toxic effects on angiogenesis, embryo development, embryonic movement and phytotoxicity of the PM from TSA and CNG powered FSA. Based on high amounts of aluminum quantified during PIXE analysis of PM from TSA and FSA, different concentrations of aluminum sulfate were also tested to determine its eco-toxicological potential. The PM solution from FSA, TSA and Aluminum sulfate exhibited anti-angiogenic potential with reduction in total area of CAM. Morphological evaluation of embryos exhibited varying degrees of hemorrhages in different groups. In case of phytotoxicity screening using Zea mays, the results demonstrated that all three tested materials were equally phytotoxic at higher concentrations in seed germination(p<0.001). Aluminum sulfate proved to be a highly phytotoxic agent even at the lowest concentration examined. During the last phase, of the study, the MTT assay demonstrated a significant (p<0.001) dose dependent cytotoxic effect for TSA, FSA and aluminum sulfate on the BHK-21 cell line, establishing that the PM from FSA is a highly cytotoxic material. Mutagenicity was assessed by fluctuation Salmonella reverse mutation assay adopting TA100 and TA98 mutant strains with (+S9) and without (-S9) metabolic activation. Despite the fact that different concentrations of PM from both sources i.e. TSA and FSA were highly mutagenic (p<0.001) even at lower concentrations, the mutagenic index was higher in TSA. The chronic toxicity study revealed that chronic exposure to PM emitted from FSA and TSA resulted in peribrochiolitis, emphesema and infilteration of leukocytes in lung tissues. On the other hand liver, cardiac and kidney tissues exhibited degeneration and necrosis. The data shows that all tested materials are equally ecotoxicand if the existing trend of atmospheric pollution by auto-rickshaws is continued, air-borne metals/heavy metals will seriously affect the normal growth of local inhabitants and increased contamination of agricultural products, which will amplify the dietary intake of toxic element and could result in genetic mutation or long-term health implications. Availability: Items available for loan: UVAS Library [Call number: 1795,T] (1).

8. Production, Purification & Characterization Of Recombinant Thermostable Phytase And Its Biological Evaluation In Broiler Chicks

by Furqan Sabir (2007-VA-524) | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Dr. Ali Raza Awan.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Phytate is the principle storage form of phosphorus in plants particularly in cereal grains and legumes. Mono-gastric animals doesn’t have ability to utilize phytate as phosphorus source. The animals release the undigested phytate from body with manure that cause environmental pollution. Phytases are responsible for the hydrolysis of phytate, resulting in availability of free phosphorus for the animal. The present study deals with the production and characterization of recombinant thermostable phytase and its biological evaluation in the broiler chicks. The PCR resulted in the amplification of 1.8 kb phytase gene using the genomic DNA of Thermotoga naphthophila as template. The purified PCR product was ligated in pTZ57R/T and the ligated material was utilized for the transformation of E.coli DH5α cells. The positive clones were selected on the basis of blue white screening. The restriction digestion of plasmid DNA from positive clones using NdeI and Hind III resulted in the release insert from the vector. The purified phytase gene after restriction digestion was ligated into pET21a already restricted with the same restriction enzymes and the expression was analyzed using E.coli BL21 CodonPlus (DEL) cells. SDS-PAGE demonstrated the intra-cellular production of recombinant phytase. The conditions were optimized for the optimal production of recombinant phytase (PHYTN). The maximal production of PHYTN was recorded when the BL21 CodonPlus cells having recombinant pET21a having phytase gene were induced with 1.4 mM IPTG and 6 hours post induction incubation period. The recombinant protein was purified using various chromatographic techniques and the purified protein was utilized for characterization. PHYTN showed optimal activity at 80 °C and pH 6 in sodium acetate buffer. The enzyme was found metal dependent and presence of Fe3+ or Cu2+ showed enhancing effect on PHYTN activity. Thermostability studies demonstrated that PHYTN retains 90% residual SUMMARY 71 activity when the protein was incubated at 80 °C for 1h in the presence of 1.5 mM Fe3+. The kinetic studies of PHYTN demonstrated km and Vmax values of 50 mM and 2500 μmole/min respectively when sodium phytate was used as substrate. The characterized PHYTN was used for poultry trials to check the efficacy of the enzyme in poultry birds. The results depicted that PHYTN put significant effect on the bird weight gain, feed intake and feed efficiency ratio. Presence of 1000 IU/kg of PHYTN resulted in the weight gain in 3rd, 4th and 5th week of trials from 504.766 to 533.535 g, 767.933 to 823.733 g and 999.833 to 1120.277 g respectively when compared with the control. The study demonstrated that this recombinant thermostable phytase is suitable for poultry feed industry and its domestic production will contribute the economic availability of PHYTN for the poultry feed industry. Availability: Items available for loan: UVAS Library [Call number: 2870-T] (1).

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