1.
Isolation, Identification And Control Of Vancomycin Resistant Staphylococcus Aureus
by Fakhra Liaqat | Dr. Ali Ahmad Sheikh | Dr. Jawad Nazir | Dr. Tanveer Hussain.
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Publisher: 2013Dissertation note: Antimicrobial resistance (AMR) is an emerging issue throughout the world. Vancomycin resistant Staphylococcus aureus has been recently reported from many countries including Asian region. The failure of antibiotics diverts the focus of modern science towards the discovery and application of new alternative antimicrobial agents. Herbal plants not only known for their antimicrobial potential but are being used since centuries for the treatment of infections. This study has been conducted to isolate Vancomycin Resistant Staphylococcus aureus from wounds of hospitalized patients and these isolates were not only tested against Linezolid, Moxifloxacin, and Clindamycin antibiotics but also against the ethanolic extracts of Garlic, Mint, Coriander, Turmeric, Kalonji, Cinnamon and Cloves for their antibacterial activity.
Methicillin resistant Staphylococcus aureus were isolated from wounds and resistance to vancomycin was confirmed according to Clinical and Laboratory Standards Institute recommended method. Minimum inhibitory concentrations of selected antibiotics and selected plant extract was determined by broth microdilution method followed by the measurement of minimum bactericidal concentration by culturing on agar plates.
In current study 104 S. aureus isolates were recovered from 150 wound exudates samples. Resistance to methicillin was shown by 49.04% isolates. Final results yielded, 22 vancomycin intermediate and 5 vancomycin resistant S. aureus strains. Vancomycin resistant isolates were tested for susceptibility against selected antibiotics and ethanolic extracts of plants. Almost all the isolates displayed susceptibility to all three antibiotics and the plant extracts. The data was analyzed statistically by chi square test and one way ANOVA using Statistical Package for Social Science (SPSS) 18.0 software. This study was helpful to find out the effective antibiotics against Vancomycin Resistant Staphylococcus aureus. Plant extracts which were found effective are the best alternate to the conventional antibiotics without having any drawback of antibiotic resistance development.
Availability: Items available for loan: UVAS Library [Call number: 1615,T] (1).
2.
Physicochemical Factors Affecting Infectivity Of Pesti Des Petits Ruminants Virus
by Kinza Khan | Prof. Dr. Khushi Muhammad | Dr. Jawad Nazir | Dr. Mutti-ur-Rehma.
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drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1636,T] (1).
3.
Isolation And Characterization Of Auxin Producing Bacterial Strains From Plant Rhizosphere
by Kanwal Aziz | Dr. Jawad Nazir | Dr | Prof. Dr. Tahir Yaqub.
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Publisher: 2013Dissertation note: Auxins are a class of plant hormones or plant growth substances. Auxins have a cardinal role in the regulation of many growth and developmental processes in the plant. Soil samples were serially diluted and screened for auxin production by Salkowski method. Isolates that have the ability to produce auxin were identified by culture characters, morphology, and bio-chemical profile. From 150 isolates, 04 bacteria were selected (AUX-36, AUX-53, AUX-137, and AUX-142). The bacteria were identified by following the flowcharts described in “Berges Mannual of Determinative Bacteriology”, 9th addition. These isolates were identified as Bacillus megaterium, Escherichia coli, Klebsiella pneumonia and Bacillus marinus respectively. Next, different physical and chemical parameters for growth of bacteria and auxin biosynthesis were optimized. For the optimization of bacterial growth OD values of the culture broth (at wavelength 600nm) was taken by spectrophotometer. To estimate the amount of auxin (ìg/ml) produced in the culture broth; a standard curve (concentration of auxin ìg/ml at x-axis and OD value at y-axis) was prepared by using commercially available auxin.
The optimum conditions for growth and auxin production by AUX-36 was found to be pH 7, 0.98% osmotic pressure at 37 °C after 72 hours of incubation. If the medium is supplemented with 0.1 and 1.0% glucose, sucrose and peptone then it increased the bacterial growth which ultimately increased the auxin concentration in the broth medium. The growth and auxin
Production by AUX-53 was 0.98% NaCl concentration at 37 °C after 72 hours of incubation. The optimum pH was found to be 7 but it showed good growth at acidic as well as alkaline pH. The addition of glucose and sucrose in the growth medium increased the growth as well as auxin production. The optimum conditions for the growth of AUX-137 were as follows: pH=7, 0.98% osmotic pressure, temperature 37 °C. However the isolate had good growth at 28 °C and 2% NaCl concentration as well. The bacterial cell density and auxin increased with incubation time up to 72 hours. The isolate produced highest concentration of auxin under the same conditions. Similarly, the cell density and auxin increased with the increasing concentration of glucose in the growth medium. Sucrose increased the auxin only in the culture filtrate. While the bacteria AUX-142 showed highest growth as well as auxin production at 42 °C after 72 hours of incubation. The optimum pH and osmotic pressure was found to be 7 and 2% respectively. The cell density and concentration of auxin increased with the increasing concentration of peptone in the growth medium. Addition of tryptophan (1-2%) increased the auxin concentration in the culture supernatant of all isolates.
Next, the seed germination test and plant pot experiment were performed of selected isolates to observe the effect of bacterial inoculation on wheat plants. In seed germination test treatment of seeds with AUX-36, AUX-53 and AUX-142 significantly increased the root length and number of root hairs as compared to non-treated seeds. In plant pot experiment comparison of various growth parameters of inoculated plants with non-inoculated plants revealed the improvement in plant growth by bacterial inoculation.
Availability: Items available for loan: UVAS Library [Call number: 1684,T] (1).
4.
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.
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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).
5.
Preparation & Evaluation Of Combined Pre & Fmd Vaccines For Small Ruminants
by Zanira Shakoor | Dr. Jawad Nazir | Prof. Dr. Khushi Mohammad.
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Publisher: 2013Dissertation note: PPR is an acute, febrile and contagious viral disease of small ruminants caused by Morbillivirus. Similarly, FMD is another highly contagious viral disease of cloven hoofed animals that affects more than 33 species of domestic and wild animals. Although disease severity is relatively higher in large ruminants but small ruminants might also play an important role in the epidemiology and transmission of FMD. Mass vaccination is the most effective mean to control viral diseases like PPR and FMD in small ruminants. The present study was designed to evaluate the immune response of goats to various monovalent and bivalent as well as adjuvant and non adjuvant PPRV and FMD "O" virus vaccines. A total of nine groups of animals each comprising 5, were inoculated with various formulations of the vaccines. Serum samples were collected at the beginning and at 1, 2, and 3 months post vaccination. Mean neutralizing antibody titers (MNA) against PPRV and MNA along with complement fixing (CF) antibody titers against FMD "O" in the serum samples were measured to test the efficacy of the vaccines.
Live attenuated wet, gel and oil based PPR vaccines induced 229.2 (±112.79), 253.6(±83.05), and 424 (±182.06) MNA titers in the goats after 3 months of vaccination. Similarly, animals inoculated non-adjuvant, gel based, and oil adjuvant FMD "O" vaccines developed 20(±4.35), 186.2 (±65.39), and 285.8 (±63.80) MNA and 0.4 (±0.894),22.4(±8.76) and 25.6(±8.7) CF antibody titers at 3 months post vaccination (PV). Bivalent (PPRV and FMDV) non adjuvant vaccine induced 249.6(±3.130) and 27.2(±14.53), gel based vaccine induced 306 (±99.82) and 296.2 (±58.21) while the oil based vaccine provoked 417.8 (±141.56) and 286 (±97.13) MNA titres against PPRV and FMD "O" virus respectively, at 3 months PV. Results of present study demonstrate that live attenuated PPRV vaccines can induce better immune response in comparison to killed FMD "O" virus vaccine. Addition of adjuvants is helpful to enhance the immunogenicity of the added antigens. While adjuvant bivalent vaccines containing PPRV and FMD "O" virus can effectively provoke equivalent antibody response against both of the immunogens in the vaccine.
Antibody titee in response to Bivalent vaccine of PPR and FMD seotype"O" containing oil as an adjuvant was superior to gel adjuvant and non adjuvant bivalent vaccines. There was poor immune response of goats to non adjuvant FMD"O" vaccines.
Availability: Items available for loan: UVAS Library [Call number: 1698,T] (1).
6.
Histopathological & Immunohistochemical Studies In Experimentally Infected Commercial
by Raoof Aslam | Dr. Asim Aslam | Dr. Jawad Nazir | Dr. Yasin Tipu.
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drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1704,T] (1).
7.
Isolation Identification & Molecular Based Investigation Of Bovine Rotavirus
by Ambreen Masood | Prof. Dr. Tahir Yaqoob | Dr. Jawad Nazir | Dr. Sehrish Firyal.
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drama
Publisher: 2013Dissertation note: Livestock is an important part of the economy of Pakistan. Calf's diarrhea due to group A bovine rotavirus causes high morbidity and mortality, which results in significant economic losses to livestock. In Pakistan overall prevalence of bovine rotavirus infection is 2.6%. As Pakistan is a developing country, survival of calves is really important to produce milk, meat and hides for propagation of livestock. The aim of current study was to isolate bovine rotavirus from faecal samples of diarrheic calves by antigen capture ELISA and molecular investigations. So, it was helpful to check the prevalence of bovine rotavirus in Lahore district.
This study will be a milestone for better treatment strategies of calf diarrhea problem. It will also pave the way for better vaccine development strategies to cure the disease. A total of 100 diarrheic faecal samples of cattle and buffalo's calves less than three months of age were collected from Lahore district. Rotavirus screening was done by direct sandwich ELISA by using commercial Rotavirus detection kit (Cypress Diagnostics, Belgium). ELISA confirmed 12 samples to be positive for bovine rotavirus. Among 12 positive samples, 7 were found positive in buffalo calf and 5 in cattle calf. After RNA extraction and cDNA synthesis, the PCR was done for amplification of VP4 gene of all ELISA positive bovine rotavirus samples. But only 5 samples (3 buffalo calf samples and 2 cattle calf samples) give desired product of 880 bp of VP4 gene. After sequencing and bioinformatics analysis, phylogenetic tree was constructed. It is evident that Pakistani bovine rotavirus VP4 gene (BRV/QOL/13) has maximum identity of 98% with Indian bovine rotaviruses VP4 gene.
Availability: Items available for loan: UVAS Library [Call number: 1707,T] (1).
8.
Association Between Numbers Of Ovarian Follicles And Fertility In Nili-Ravi Buffaloes
by Qaisar Shahzad | Dr. Amjad Riaz | Dr. Jawad Nazir | Dr. Mian Abdul Sattar.
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Publisher: 2013Dissertation note: 6.1: Back Ground:
The water buffalo (Bubalus bubalis) is an important animal of livestock species. It has an essential role in the economy of developing countries. Buffalo is being used as dairy, beef and draught purpose. Despite so many qualities, the reproductive potential of the buffalo is lower than cattle. One of the reasons for low reproductive potential is smaller ovaries and low number of ovarian follicles. Due to these reasons superovulation and embryo transfer has not been much successful in buffaloes. There is no tool through which reproductive performance of buffaloes can be phenotyped. Antral follicular count can be a tool on the basis of which reproductive performance of buffaloes can be phenotyped
6.2: Hypothesis:
Higher the antral follicular count higher will be the fertility in Nili-Ravi buffaloes.
6.3: Methodology:
Ten Nili-Ravi heifers were used in the study to measure repeatability of antral follicular. Each heifer was scanned on alternating days from day 1 of estrous cycles to day 9 of the successive estrous cycle. Antral follicles were counted in different follicular waves and repeatability of antral follicles was assessed. In the 2nd experiment of the study 10 heifers were used to count antral follicles from day 1 to day 10 of the estrous cycle, on the basis of antral follicular count animals were divided into three groups, Low (1-4), Intermediate (5-8) and High (?9).Blood sampling was done on 7th day of estrous cycle to measure progesterone concentration of 3 animals from each group and an association was developed between antral follicular count and progesterone concentration. In the 3rd experiment, 25 Nili-Ravi buffaloes were used to antral follicles from day 1 of estrous cycle to day 10 of estrous cycle. , on the basis of antral follicular count animals were divided into three groups, Low (1-4), Intermediate (5-8) and High (?9). In the next heat animals were artificially inseminated and were checked for pregnancy on 36th and 60th day by using ultrasound. Animals pregnant on 60th day were considered as pregnant. After that association was developed between antral follicular count and fertility.
6.4: Results:
Buffalo can be phenotyped on the basis of antral follicular count. Follicular counts are highly repeatable in different follicular waves of same estrous cycle (0.83) and different estrous cycles (0.85) within individual animals. Follicular counts are highly positively correlated (0.91) with progesterone concentration. Higher the antral follicular count, higher should be the progesterone concentration. Antral follicular count is highly positively associated with (0.99) with fertility. Higher the antral follicular count higher will be the fertility.
Availability: Items available for loan: UVAS Library [Call number: 1734,T] (1).
9.
Standardization Of Multiplex Rt-Pcr For The Diagnosis Of Avian Influenza (H9) Virus Newcastle Disease Virus (Ndv)
by Asad Shahzad | Dr.Asim Aslam | DR. Jawad Nazir | DR.Raheela Akhtar.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1917,T] (1).
10.
Study On Micobiological Quality Of Water Supplied To Poultry Birds From Tube Wells Water Pumps Drilled Up To Varying Bore
by Sidra Moqddes | Prof. Dr. Masood rabbani | Dr. Jawad Nazir | DR.Yasin tipu.
Material type: ; Format:
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Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1922,T] (1).
11.
Role Of Water Chemistryand Stabilizers On The Infectivity Of Virus In Live Neqcastle Disease Virus Vaccine
by Sahrish tariq | Dr Jawad Nazir | Prof. Dr. Masood Rabbani.
Material type: ; Format:
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Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1927,T] (1).
12.
Microbiological And Physiochemical Analysis Of Drinking Water From Human And Veterinary Hospitals
by Kiran batool | DR. Arfan ahmad | Dr Hassan mushtaq | DR. jawad nazir.
Material type: ; Format:
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1958,T] (1).
13.
Isolation Characterization And Optimization Of Potential Probiotic Bacteria From Poultry Droopings
by Muhammad Hashim khan | Prof. Dr. Aftab ahmad anjum | Dr. Jawad nazir | Prof. Dr. Mansur-ud-din.
Material type: ; Format:
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1991,T] (1).
14.
Screening Of Indigenous Microalgae For Antimicrobial Activities And Growth Optimicrobial For Mass Production
by Imran hanif | Prof. Aftab ahmad anjum | Dr. Jawad nazir | Ms. Sehrish firyal.
Material type: ; Format:
print
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2015,T] (1).
15.
Effect Of Rbst Treatment On Antral Follicular Count Plasma E2 P4 Profile And Esteus Behavior In Postpartum Nili Ravi Buffaloes
by Sadia naz | Dr. Amjad riaz | Dr. Jawad nazir | Prof. Dr. Nasim Ahmad.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2055,T] (1).
16.
Ameliorating Effects Of Acetic Acid On Performance And Performance And Histopathological Parameters In Broiler
by Rukhshanda Ramzan | Dr. Gulbeena Saleem | Dr. Jawad Nazir | DR. Yaseen Tipu.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2107,T] (1).
17.
Follicular Dynamics During Estrous Cycle In Sahiwal Cows
by Muhammad Yasir Arfat (2007-VA-102) | Prof. Dr. Nasim Ahmad | Dr. Amjad Riaz | Dr. Jawad Nazir | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2014Dissertation note: Pakistan by default is an agricultural country. Livestock is mainstay of the farming
communities and has exclusive position in national agenda of the development. It plays an
important role in poverty alleviation and can uplift the socioeconomic condition of our rural
masses. Livestock contribution to agricultural GDP is 55.9 % and its contribution to National
GDP is 11.8 % (Anonymous,2013-2014). Total cattle population of Pakistan is 39.7 M
(Anonymous, 2013-2014). Livestock, especially cattle, play an important role in agriculture
economy of Pakistan in form of milk, meat and draught power. Milk is one of the cheapest
sources of nutrition and is beneficial for the human health in all stages of life. Despite the
nutritional importance of milk, its per capita availability and consumption is low in our country.
This inadequacy is due to suboptimal performance of indigenous dairy cattle due to lack of
modern technologies in cattle farming.
Pakistan is blessed with the finest breeds of dairy cattle such as Sahiwal, Cholistani and
Red Sindhi. Sahiwal cattle breed initially named as Montgomery breed (Bos indicus) is one of
the important breeds of indigenous cattle in Pakistan having dairy characteristics. The average
milk yield is about 1500 liters per lactation with 4% butter fat. But still its potential of milk
production is far less as compared to the exotic breeds e.g. Holstein Friesian etc. This is
primarily due to compromised feeding and management and little attention in the past for the
selection and breed improvement in Sahiwal cows. Moreover, late age at maturity and longer
calving interval (Makuza and McDaniel 1996) are major reproductive issues in Sahiwal cows.
Introduction
2
Physiology of oestrus cycle has been extensively studied in Holstein cows ( ). With the
advent of ultrasonography in early 1980’s it became possible to study follicular and luteal
dynamics during the estrous cycle in detail in Bos taurus (Fortune 1994; Lucy et al. 1992; Savio
et al. 1988; Wolfenson et al. 1995), and some beef breeds of Bos indicus cattle (Bó et al. 2003;
Figueiredo et al. 1997) and in buffalo (bubalis bubalis) as well (Warriach and Ahmad 2007).
The benefit of these studies was that the information on follicular dynamics in Bos taurus breeds
has been used to manipulate the estrous cycle in order to improve estrus synchronization
(Thatcher et al. 1993; Twagiramungu et al. 1995; Wolfenson et al. 1994) fixed time artificial
insemination (FTAI) (Pursley et al. 1997; Schmitt et al. 1996a; Schmitt et al. 1996b;
Twagiramungu et al. 1995) and embryo transfer procedure (Mapletoft et al. 1994; Roberts et al.
1994).
Sahiwal is one of the established zebu cattle (bos indicus) milk breeds of tropical and
subtropical region. It is known for its remarkable power of endurance for hot climate, resistance
to ticks and other diseases and has high producing ability under harsh environment and low cost
of maintenance as compared to the Bos indicus and Bos taurus crossbreds. Due to its promising
dairy characteristics and better adaptation to tropical environmental conditions, both the semen
and female of this breed have been exported from Pakistan and in Africa and Australia.
Differences on the reproductive characteristics between Bos taurus and few breeds of Bos
indicus cattle have been reported like luteal tissue characteristics (Pathiraja et al. 1986), Graafian
follicle (DF) diameter (Figueiredo et al. 1997) and estrous cycle duration (Castilho et al. 1996).
However, surprisingly, there has been no thorough study on the reproductive physiology of the
estrous cycle in Sahiwal cows. Therefore, the objective of the present study is to determine the
Introduction
3
follicular dynamics, luteal tissue development and regression, estrous cycle length, timing of
ovulation, estrus signs and fertility.
It is hoped that these data will be helpful for improved assisted reproductive technology
e.g. AI, ET etc. (Andrabi and Maxwell., 2007), timing of the treatment of the various hormones
(Krininger et al., 2003) and development of new technologies like fixed time A.I, estrus
synchronization, super ovulation, embryo transfer in Sahiwal cows. Ultimately, these can
increase herd reproductive, productive performance and for preservation of Sahiwal cattle breed. Availability: Items available for loan: UVAS Library [Call number: 2183,T] (1).
18.
Effect Of Temperature And Relative Humidity On The Survival Of Newcastle Disease Virus Isolates Using Germ Carrier Techniques
by Tayyeba Sohail (2009-VA-209) | Dr. Jawad Nazir | Prof. Dr. Khushi Muhammad) | Prof. Dr. Asim Aslam.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Newcastle Disease (ND) is a highly contagious viral disease that affects almost all avian species including poultry, cage and wild birds around the globe (Terregino et al. 2003; Vidanovic et al. 2011). ND is economically important disease and included as list-A disease of Office des International Epizootics (OIE) (Anonymous). Mortality of infected birds ranges from negligible to as high as 100 % depending on the pathotype of the virus involved and health status of the birds (Alexander and Manvell 2004). NDV is an enveloped virus with single stranded, non-segmented, negative sense RNA genome (Makoui et al. 2013). The virus belongs to Avulavirus genus of Paramyxoviridae. There exist only one serotype of NDV designated as avian paramyxovirus-1 (Kapczynski et al. 2013) however, different virus strains do vary in their pathogenicity. There are 3 pathotypes of NDV; velogenic (highly virulent), mesogenic (moderate virulent), and lentogenic (mild virulent) based upon diseases producing potential and severity of signs in the infected birds (de Leeuw and Peeters 1999).
NDV is primarily transmitted to the susceptible birds through aerosol and fecal oral route (Martin 1992). Infected birds secrete high amount of the virus in their feces, saliva, mucous and nasal secretions which might contaminate the premises. Inanimate objects or fomites are a potential reservoir of viruses outside the host and might play an important role in the transmission of pathogens (Nicas and Sun 2006). Several factors can influence the survival of viruses outside the host (Sobsey and Meschke 2003; Weber and Stilianakis 2008; Stallknecht and Brown 2009). A number of studies show that respiratory pathogens can survive from hours to months on fomites (Abad et al. 2001; Kramer et al. 2006). Certain physical factors like temperature, humidity, pH, salinity, exposure to ultraviolet (UV) rays etc drastically affects the
Introduction
2
virus persistence in the environment. Effect of such physical insults is more pronounced on enveloped viruses than non-enveloped ones (Mbithi et al. 1991; Schaap et al. 2012; Tuladhar et al. 2012). High humidity and temperatures not only reduces the survival of influenza viruses on contaminated surfaces but also modulates their transmission to the susceptible birds (Shaman and Kohn 2009; McDevitt et al. 2010; Paynter 2014). Similarly lower temperature and less humidity promote the survival of NDV in the environment (Dat and Chuc 1985; Kournikakis et al. 1988).
ND is endemic in Pakistan but since last few years several new virus strains are circulating in commercial and rural poultry of the country (Munir et al. 2012; Shabbir et al. 2013). Central Punjab region is densely populated with commercial poultry and serve as disease epicenter every year. It has been observed that the disease outbreaks usually start in December, attain peak in the late winter and spring season, start decline in June and disappear in the rainy season. Apart from several other contributing factors, environmental survival of the viruses might contribute to the disease outbreaks. Availability: Items available for loan: UVAS Library [Call number: 2227-T] (1).
19.
Anticoccidial Activity Of Aloe Vera (Qawar Gandal) In Broiler
by Hafiz Atif Munir (2005-VA-102) | Dr. Muhammad Lateef | Dr. Haroon Akber | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: There are various species of Eimeria that cause coccidiosis. This disease is considered to be one of the most significant and prevalent diseases across the world. This disease reduces the production and causes high morbidity and death rates, on the other hand for controlling this eimeriosis, one has to adopt preventive therapy in the form of anticoccidial drugs in combination with occurrence of natural immunity in birds. Coccidia cause infection in intestine and multiply in gut and belong to protozoan parasites. These protozoan coccidian cause chicken Eimeriosis which is considered one of the most significant diseases, that cause severe economic losses across the world. For poultry control houses, coccidiosis is considered one of the most common (Hamidinejat et al., 2010).
As oocysts of coccidia are very unique so they spread from one poultry farm to another poultry farm and for dissemination of this protozoan, parasitic disease management plays a pivotal role. Now it’s very hard to keep chickens safe from these parasitic protozoa particularly in the control poultry houses where high production is required. In poultry house litter, oocycts multiply readily. But there are some factors which affect the number of these protozoa like bacteria or some environmental factors like liberation of Ammonia gases and under these factors, Eimeria start to diminish after three weeks (Williams, 1995).
Caecal and intestinal forms are the two important forms of avian coccidiosis. Diarrhoea and excessive caecal haemorrhages are the main symptoms of caecal coccidiosis and its causative agent is Eimeria tenella (Gardinar 1955). In prevention, surveillance and control of coccidiosis specific diagnosis plays a pivotal role. Eimeria oocysts from the faeces of infected chickens have been detected or enumerated by conventional methods and pathological lesions resulted by Eimeria infection and oocyst structure can also be determined by employing traditional methods (Long and Joyner 1984). But because of the involvement of multiple species of Eimeria in coccidial infection these approaches cannot be realistic as there can be overlapping among various sizes of oocysts (Long and Joyner 1984).
There are six species of Eimeria which cause great economic loss in chickens and also these species are highly host specific. High mortality rate and morbidity rate due to bloody enteritis and production losses are reasons of great economic. Despite management now a days anticoccidial drugs have been used in chicken feed to minimize the effect of this disease. E. tenella, E. necatrix, E acervulina, E. maxima, E. brunetti , E. mitis and E. praecox are the seven species of Eimeria which are considered as main causative agents of coccidiosis in chicken (Arabkhazaeli et al. 2011).
However, control of coccidiosis can be achieved by producing species-specific natural immunity along with anticoccidial drugs (Shirley et al. 2004), but it’s also evident from the fact that drug resistance has been increasing due to excessive use of anticoccidial drugs (Williams 1998). Pathological lesions, host and protozoan characteristics are very important for diagnosis of coccidiosis. But one cannot identify the Eimeria species accurately as analysis of these characteristics requires high expertise.
Good management plays key role in controlling coccidiosis which includes proper ventilation, dry litter, drinkers and feeders in good clean condition and appropriate density of stock (Jordan 1995, Gross 1985). Size and morphology of parasite at various stages of life cycle plays an important role in identification of Eimeria and site of enteric pathological lesions. In complex infection it’s very hard to identify the species showing same features (Long and Reid 1982).
The increasing resistance of avian coccidiosis to anti-coccidial drugs currently used by poultry industry together with the requirement for drug and the production systems which should be antibiotic free, it’s now very necessary to go for the new and advanced methods to prevent from this disease. Therefore, scientists started to work on medicinal use of herbal products to control this eimeriosis. In past for treatment of various human and poultry ailments the natural herbal products have been effectively used several times. Because of the anticoccidial nature and antibacterial effect, Aloe vera could be served as a valuable alternative for coccidiostats as a medicinal plant. Previously aloe gel has been used for multipurpose like as an antibiotic, for healing of wounds, for anti-inflammatory effects, for anti Eimerial response and as an anti-ulcer agent. It was also reported that in broilers it increases immunity by enhancing the number of microvilli (Jinag et al., 2005). In Asia Aloe gel is considered one of the most common and easily available country medicines to get effective deliberate results. Therefore, the current study planned to cope coccidial diseases in broilers by serving Aloe vera as an effective weapon.
The present study therefore planned to discover the anti-coccidial effects of Aloe vera in broilers by using its powdered form, aqueous and methanol extracts.
Availability: Items available for loan: UVAS Library [Call number: 2231-T] (1).
20.
Efficacy of Various Disinfectants Against Newcastle Disease Virus Isolates In Relation To Different Temperatures
by Momena Habib | Dr. Jawad Nazir | Prof.Dr.Aftab Ahmed Anjum | Dr. Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Newcastle disease (ND) is an infectious viral disease of birds and has been considered as one of the most significant diseases of poultry and other avian species around the globe (Aldous and Alexander 2001). The disease is endemic in Pakistan and causes major economic losses to poultry industry each year (Rehman H et al. 2013). Although incidence of ND in broiler and layers remains higher during the course of year, it reaches its plateau during seasonal stress (January-February and June-July). One report shows that during September 2011 - January 2012, the diseases has killed 45 million birds and resulted into a loss of approximately 65 million US $ to the country (Anonymous). ND virus (NDV) is an enveloped, single stranded RNA virus and grouped into the genus Avulavirus within family Paramyxoviridae(Calibeo-Hayes et al. 2003). Isolates of NDV may be categorized into three main pathotypes (lentogenic, mesogenic and velogenic) depending upon the severity of disease in chickens (Seal et al. 2000).
NDV is most commonly transmitted through fecal-oral and respiratory route provided that birds are in close contact (Alexander 1988). Infected birds excrete large amount of virus in their feces (Calibeo-Hayes et al. 2003). Virus is present in most tissue secretions of acutely infected birds even 24 hours prior to appearance of clinical signs. Under certain conditions, NDV can survive up to 20 days in fecal material (anonymous) and presence of organic matter might enhance the virus survival rates (Clarke et al. 1956). In order to control the spread of disease, decontaminate the infected premises is of prime importance. Strict biosecurity measures in conjunction with mass vaccination programs are also employed for the control of the disease. In past few decades, implementation of extensive vaccination programs in commercial poultry has reduced the
Introduction
2
disease incidence to some extent in Pakistan. However, this might also led to the generation of novel genotypes under high immune pressure(Miller et al. 2009). Disinfection of farm premises plays significant role to break the disease cycle of microbial pathogens(Fawzia1 et al. 2013). Such procedures gain more significance on the farms that experience disease outbreaks. Dry cleaning, burning and fumigation followed by chemical disinfection is routinely practiced in Pakistan. Although, physical method are helpful to reduce the pathogen load on farm premises but complete disinfection cannot be achieved without the use of chemical disinfectants. Commercial disinfectants employed in Pakistan are grouped into oxidizing agents, aldehydes, quaternary ammonium compounds, and phenolic preparations. The products containing these ingredients are available either alone or various combinations under different brand names. (Stringfellow et al. 2009).
All of the disinfectants do vary in their disinfection power but to achieve proper disinfectants appropriate interaction time is necessary. Under clean conditions such interaction time is low but longer contact times are needed in presence of organic matter. Some studies show that environmental temperature influences the interaction time of disinfects with matter/surfaces to achieve proper disinfection. In case of chlorine disinfection of adenovirus inactivation time was increased 2-3 times when temperature was lowered to an extent of 10°C (Clarke et al. 1956). Similarly, virucidal activity of some disinfectants including aldehyde, chlorine and iodine compounds against infectious bursaI disease virus (IBDV) was also decreased at lower temperatures (Benton et al. 1967). Availability: Items available for loan: UVAS Library [Call number: 2226-T] (1).
21.
Isolation Of Surface Antigen 1 Gene Of Toxoplasma Gondii And Its Cloning In The Expression Plasmid
by Farooq Riaz (2008-VA-231) | Dr. Muhammad Imran Rashid | Prof. Dr. Kamran Ashraf | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Toxoplasma gondii is an obligate intracellular protozoan parasite which comes under the classification of phylum Apicomplexa, subclass Coccidiasina (Cornelissen et al. 1984). Toxoplasmosis is one of the more common parasitic zoonoses world-wide caused by Toxoplasma gondii which is a facultatively heteroxenous, polyxenous protozoon that has developed several potential routes of transmission within and between different host species (Tenter et al. 2000). It is the most important source of toxoplasmosis in humans and animals, with cat as definite host and warm-blooded animals as intermediate host (Frenkel et al. 1970). It was first described by Nicolle, Manceaux and Splendore in 1908 from rodents Ctenodactylus gondii (Black and Boothroyd 2000).
Toxoplasmosis is a worldwide parasitic disease and it is estimated that about one-third total population of the world is seropositive for Toxoplasma gondii (Tenter et al. 2000). Prevalence of infection varies between countries, geographical areas and ethnic groups living within a specific region. In Humans, infection rates range from 50% to 83% in Brazil (Tenter et al. 2000; Dubey et al. 2012). Seropositivity of Toxoplasma gondii in China is about 8% with continuously increase while in USA its 10-15%, 50-70% in France and 20% in UK (Dubey and Jones 2008; Zhou et al. 2008; Jones et al. 2009). Prevalence of toxoplasmosis is higher in males (79%) as compared to females (63.4%) and the age dependent sero-prevalence reaches >92% in age group of 40 to 50 (Coêlho et al. 2003).
Transmission occurs through the ingestion of contaminated vegetable /water with oocysts, as well as the ingestion of contaminated raw/undercooked meat with tissue cysts (Gajadhar et al. 2006). Transmission may also occurs by ingestion of sporulated oocysts, or bradyzoites within cysts present in the tissues of numerous food animals (Esteban-Redondo et al. 1999). In humans, transmission of Toxoplasma gondii happens mainly by eating raw or undercooked contaminated meat, raw cow’s milk and birds eggs, swallowing oocysts dis-charged in feces of infected cats, inoculation of trophozoites through the skin, or by inhalation (Wallace 1971; Wallace 1973; Bannister 1982).
In humans, mostly infections (congenitally or post-natally acquired) are asymptomatic. Congenital infection occurs only when a woman becomes infected during pregnancy. Congenital infections acquired during the first trimester are more severe than those acquired in the second and third trimester (Desmonts and Couvreur 1974). The main clinical signs associated with toxoplasmosis are anorexia, weight loss, lethargy, dyspnea, ocular signs, pyrexia, vomiting and diarrhea, jaundice, myositis, encephalitis and abortion. Humans become infected when they ingest the toxoplasma at infective stages (oocysts and tissue cysts) found in some cat feces and in raw meats.
In addition to being hazardous to livestock animals, the T. gondii infection is also important due to its zoonotic implications (Jittapalapong et al. 2005). Congenital abnormalities in humans, such as microcephaly, hydrocephaly, chorioretinitis, convulsion, cerebral calcification, epilepsy, blindness, deafness, and mental retardation may occur if the mother acquires infection during pregnancy (Jones et al. 2003). In addition to congenital anomalies, T. gondii also causes severe neuropathologic infections in immuno-compromised hosts, such as AIDS and cancer patients receiving chemotherapy (Del Valle and Piña-Oviedo 2005).
Seroprevalence studies of T. gondii among domestic animals in South-Western Pakistan has indicated considerable prevalence (25% in cattle, 2.5% sheep) (Zaki 1995) and suggesting potential transmission to the human community. Small scale study in urban area of Rahim Yar Khan (Punjab), Pakistan has revealed that the overall prevalence of toxoplasmosis in food animals is 19% (Ramzan et al. 2009). Another study has already been published that untreated patients with leprosy in Pakistan have shown significant seroprevalence (29.6%) of antibodies against T. gondii (Hussain et al. 1992).
Vaccine against toxoplasmosis is not available yet with one exception (“Toxovax” for sheep). Vaccine against T. gondii in animals used for human consumption may block the possible transmission to humans (Bhopale 2003). SAG1, among one of the major antigenic components of Toxoplasma gondii is a major surface antigen identified on the surface membrane of this parasite using a monoclonal antibody (Handman et al. 1980). SAG1 is an important surface antigen, expressed by tachyzoite form of T. gondii and is a putative candidate for vaccine and diagnostic against toxoplasmosis (Sharma et al. 1983; Godard et al. 1990). Immunization with SAG1 adjuvanted with saponin Quil A or incorporated in lysosomes provided total protection after challenge (Bülow and Boothroyd 1991; Khan et al. 1991). SAG1 is single copy gene with no introns (Burg et al. 1988), regulates both humoral as well as cellular Th1 immune responses (Liu et al. 2008) and is powerful candidate for vaccine against toxoplasmosis. SAG1 is a potent candidate of diagnostics for detection of serum antibodies against toxoplasmosis in Man and animals (Abu-Zeid 2002).
Availability: Items available for loan: UVAS Library [Call number: 2258-T] (1).
22.
Molecular Diagnosis Of Trypanosomiasis In Pet Dogs Of Lahore
by Muhammad Asif (2007-VA-460) | Prof. Dr. Kamran Ashraf | Dr. Nisar Ahmad | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Trypanosomaevansiis a protozoa that causes surrain a wide variety of mammals. It is widely reported in adult dogs (Rashid et al. 2008; Defontis et al. 2012). Trypanosoma evansiis utmostcommonlyexisting trypanosome in animals. It is a salivarian pathogen (Hoare. 1972). Stomoxyand Tabanidssppare menifested as mainvectors universally. Oral spreadis also reported in both wild and domestic animals (Adams and Lionnet. 1983). Since 2008, surra became obligatory not only in horses, because it has been considered as a multi-species disease by the OIE (OIE. 2008; Salim et al. 2011). Surra usually follows an acute course of infection in dogs, though it is sporadically prevalent (Ravindran et al. 2008). Outbreaks of canine trypanosomiasis have been reported in India, Iran,Brazil, and South America (Herrera et al. 2004; Morteza et al. 2007; Umezawa et al. 2009). Trypanosomaevansionly has been reported from subcontinent (Ravindran et al. 2008).
Causative agent for American trypanosomiasis is Trypanosoma cruzi (T. cruzi) and African trypanosomiasis (surra or sleeping sickness) whose causative agent isTrypanosomaevansi(T.evansi).These are two forms of canine trypanosomiasis.It was originally an enzootic disease mingling in mammals and birds, which served as areservoir. The disease became zoonotic due to interaction between rural populations and natural foci, which are the results of biologicalinequity (Johns et al. 2000). Peracute to acute infections due to trypanosome result in high temperature, hemorrhages in the mucosal and serosal sides. Anemic condition of the patient is produced due to loss of RBCsfrom circulationby the mononuclear phagocytic system which is the cardinal feature of trypanosome infection.In chronic infections, anemia may be resolved due to little parasitic load in blood at capricious degrees. (Urquhart et al. 2002).
Some note able signs may compriseedema of throat and head, Blindness due corneal opacity, Temperature and anorexia. Larynx may alter the voice of the dog due edema, which can complicate with rabies. Infected dogs are considered as a risk factor in household spread of the Chagas disease in humans (Cohen and Gurtler.2001).
In native animals, dogs have the flankingassociation with humans; they may assumedconsiderable epidemiological importance in the perspective of public health and zoonosis. In humans,T.evansiproduce chronic pathological changes, which includes congestive cardiac insufficiency, finding of which isproblematic and can be unexploited due to multisystemic nature of the infection; it increases the need for epidemiological and experimental support. Moreover, causative agent of trypanosomiasis must beconfirmed by laboratory analysis, which can make availablesignificantprovisionwhen using suitable techniques, suitable reagents, and subsequent good laboratory practices (Eloy & Lucheis. 2009).
There has been development of several compounds with value against canine trypanosomiasis, however none of these products have been produced in a large commercial scale or even accessible in the market. The apparent inaccessibility of new trypanocides in the market have continued a great challenge to the treatment of the infection. Diminazine aceturate dose of 3.5 mg/kg in T.congolense infection; 7 mg/kg inT. brucei andT. evansi(Aquinos. 2007) has shown efficacy when used to treat canine trypanosomiasis.However, treatment does not provide satisfactoryresults but only sustained the life of the dog for some reasonable period (Amora. 2004).
Dogs were vaccinated with a fixed T.rangeli against canine trypanosomiasis recently (Basso et al. 2007). Experimental infections of the vaccinated dog produced disease of low parasitaemia apparently from vaccine induced immunity. Furthermore, feeding of the vaccinated dogs with the nymph stage of triatomine reduced the rate of infection in the bugs. Since dogs are the reservoir of Chagas disease in man, advances in this area could reduce the rate of infection of kissing bug which will in turn aid in the control of the disease in man (Basso et al. 2007) are necessary to establish the diagnosis.Sensitivity of direct parasitological examination is directly related to parasitic burden, biological material.
The diagnosis oftrypanosomiasis is based on combination of comprehensive clinical inspection, appropriate sample collection, sample size, suitable diagnostic tests and suitable conduction of tests and logical interpretation of results. In canine trypanosomiasis where disease prevalence is great, some tests of low diagnostic sensitivity may suffice (OIE. 2008). Parasitological diagnosis could be made by microscopic inspection either of blood, lymph node or CSF of infected dogs (François et al. 2005).
Pet dogs have been the companion of human being since ages, and shares the environment and belongings. Trypanosoma is found in dogs causing health problems effecting their routine activities. Rapid and accurate diagnosis of the infection is must to do for better therapeutic approach and early recovery of the animal. PCR is gold standard test for the molecular diagnosis of disease leading to quick diagnosis, early recovery and cost saving. So, regarding to disease importance and dogs domestications which is increasing day by day in and around Lahore area, we have focused this species to determine the Trypanosoma evansistatus in dogs in this area. This whole study is based on two diagnostic techniques i.e. screening through microscopic examination and confirmation of these samples via PCR with details regarding age, sex and breed association with the disease.
Availability: Items available for loan: UVAS Library [Call number: 2259-T] (1).
23.
Efficacy Of Surface Disinfectants Against Bacterial Pathogens On Experimentally Contaminated Pathogens
by Sana Ahmed (2009-VA-241) | Dr. Jawad Nazir | Dr. Amir Ghafoor | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Surface disinfection plays a key role in the prevention, control and reduction of many communicable infections as contaminated surfaces are the major source for transmission of microorganisms. Five types of surface cleaning products (Astonish Germ Clear, Cif Floor Cleaner, Dettol Surface Cleaner, Finis Floor Cleaner and Lysol Surface Cleaner) were evaluated for antibacterial activity against three bacterial cultures (E. coli, K. pneumonia, S. aureus) through Quantitative non-porous Surface Carrier Test derived from EN 13697.
Efficacy testing was performed through surface contamination techniques. Glass slides surfaces were artificially contaminated followed by recovery of the bacteria through vortex method both before and after the application of product for 5 minutes. Each of the experiment was repeated thrice and microbicidal effect (ME) values after the application of each product were calculated. Residual antimicrobial activity of the surface cleaning products was measured by applying the working solution of disinfectant on contaminated glass surfaces. Exposure time was given to the test surfaces, after each set exposure time the surfaces were treated to recover the microorganisms. Viable count from the eluant was calculated by serial dilution spread plate method. Each of the experiment was repeated thrice to find out the residual antimicrobial effect of the disinfectant products.
ME values of the Finis Floor Cleaner ranged from 4.03 to 5.14, which was the maximum value among all surface cleaning products used against E. coli. Highest ME value against S. aureus and K. pneumonia was shown by Astonish Germ Clear. The ME values ranged from 4.99 to 5.10 against S. aureus and 5.34 to 6.99 against K. pneumoniae. Finis Floor Cleaner was proved to be of maximum efficiency against E. coli where as Astonish Germ Clear was most effective against Staph. aureus and K. pneumonia. The mean log10 CFU values recovered from disinfectant treated
Summary
49
surfaces when they were exposed to the environment for different time periods of five minutes, six hours and 24 hours are 5.62 to 7.94, 4.17 to 6.35 and 7.16 to 10.25 respectively. The results indicated that the microbial count was reduced significantly at interaction time of five minutes, then at six hours the count was further reduced by Astonish Germ Clear, Cif Floor Cleaner and Dettol Surface Cleaner i.e. these surface cleaners were able to maintain their antimicrobial activity upto six hours. When the exposure to environment further increased to 24 hours, the microbial count started to increase, hence none of the disinfectants has shown antimicrobial activity upto 24 hours. This indicates that significant microbial count can be achieved within the interaction time of five minutes to six hours. Loss of antimicrobial activity upto 24 hours is probably because the active ingredients of cleaning agents get degraded during long interaction time.
Present study emphasizes that the surface disinfection process must be repeated at regular intervals. Regular and timely use of surface cleaning agents must be considered as a crucial measure in controlling disease transmission rates. Availability: Items available for loan: UVAS Library [Call number: 2294-T] (1).
24.
Prevalence Of Newcastle Disease In Backyard Poultry In District Mardan
by Muhammad Saeed (2013-VA-439) | Dr. Mamoona Chaudhry | Dr. Abdul Sajid | Prof. Dr. Mansur Ud Din Ahmad | Dr. Jawad Nazir.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Newcastle disease (ND) is very important viral diseases of poultry industry in the rural areas of Pakistan It is caused by Avian Paramyxovirus serotype 1 (APMV-1) of the genus Rubulavirus belonging to the family Paramyxoviridae. The outbreaks of ND are usually associated with various factors e.g. confinement of birds, mode of disposal of diseased birds, cadavers and poultry fecal matter; dry seasons in the dry zones just before the rains; wind conditions; short irregular temperature changes and the refilling of farms with chickens from the markets. The present study was conducted in randomly picked 30 clusters in three Union councils of Tehsil Takht Bhai District Mardan to investigate the seroprevalence of Newcastle Disease virus and its potential risk factor in non-vaccinated chicken raised under backyard management system. Serum were observed through Haemagglutination inhibition test for the confirmation of prevalence of Newcastle Disease. 165 were found seropositive and 45 were seronegative (antibody titres of 4 or less) for ND out of 210 sera samples. Overall weighted seroprevalence was found as 76.836%, 95% Cl (66.238-87.433) using R software. This means that NDV was circulating in backyard poultry of district Mardan, while data on risk factor were obtained through a detail predesigned questionnaire from the owner in a face to face interview translated into local language (Pushto) after taking written consent from the owner.
To identify the risk factors for Newcastle Disease seroprevalence, multivariable logistic regression were performed. The result showed that live birds market stall near houses was strongly associated with NDV seroprevalence. Source of water from both type (public water supply and street channels) were also found strongly associated. A strong association was also observed between NDV seroprevalence and water source of street channels. Result also showed that cleaning of backyard premises was a protective factor against NDV with OR < 1. Another
Summary
38
strong risk factor was live birds market stall near houses (OR 33.64, 95 % Cl: 6.49-174.28). The largest confidence interval showed less precision which could be due to less no. of samples. The identified estimate of seroprevalence of ND and its associated potential risk factor will be communicated to concerned persons through publication. Availability: Items available for loan: UVAS Library [Call number: 2292-T] (1).
25.
Isolation And Characterization Of Avian Isolates Of Lactobacilli Species And Their Antisalmonella Activity
by Anum Shaukat (2009-VA-220) | Dr. Muhammad Nawaz | Dr. Jawad Nazir | Prof. Dr. Mansur-Ud-Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Poultry industry is second largest industry of Pakistan. Poultry industry is cheaper source
of protein and provides jobs for more than 1.5 million people. It is facing several problems due
to microbial diseases. Salmonella is one of the leading causes of diseases in poultry. These are
being treated with antibiotics but misuse and overuse of antibiotics result in antibiotic resistant
strains of microorganisms. We need some alternatives for treatments. Lactobacilli are one of the
alternatives to antibiotics used as probiotics.
It has been found that the Lactobacilli of poultry origin have antimicrobial activity
against Salmonella.
Lactobacilli was isolated from the droppings, cloaca and caecum of rural poultry birds
using deMan Rogosa Sharpe (MRS) medium. The isolates were screened for anti-Salmonella
activity against S. enterica along with their properties to resist low pH and bile acids, antibiotic
sensitivity, auto-aggregation and co-aggregation. The isolates showed anti-salmonella activity
were identified using microscopic characters and biochemical profile. The isolates were
confirmed by PCR using species specific primers and sequencing 16S rRNA gene.
The data was analysed using one-way ANOVA at significance level P value <0.05 by
using the statistical software Graph Pad Prism version 5.3.
The study was conducted on a total of 60 samples including caecal swabs (n=20), cloacal
swabs (n=20) and dropping (n=20) of indigenous poultry. From these samples, five isolates were
Summary
57
selected based upon the tests performed. Isolates namely CLB-41, CLB-45, PDL-13, PDL-26
and PDL-33 showed best results. Further characterization was done by PCR and sequencing.
Availability: Items available for loan: UVAS Library [Call number: 2313-T] (1).
26.
Detection Of Brucella Species From Aborted Bovine Fetuses Using Amos Pcr And Immunohistochemistry
by Muhammad Naveed Anvar (2008-VA-310) | Dr. Raheela Akhtar | Dr. Gulbeeena Saleem | Dr. Jawad Nazir.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note:
The economic importance and public health concern of bovine brucellosis enlist it in the world top priority disease to be eliminated by WHO (World Health Organization). Bovine brucellosis is caused by three Brucella species including B. abortus, B. melitensis and B. suis. Although the literature explains that the incidence of B. abortus is higher in bovines but still we need to know the prevalence of other species particularly B. melitensis in Pakistan due to its zoonotic aspect which makes it more important for study. Unfortunately in Pakistan the status of B. abortus and B. melitensis in bovines is unknown. It is need of hour to determine the exact prevalence of B. abortus and B. melitensis in bovines for disease eradication in animals, to control its transmission in humans and to determine the reasons behind vaccine failure in bovines.
B. abortus and B. melitensis could be presents in aborted bovine samples. AMOS PCR can be better tool than immunohistochemistry for the detection of B. abortus and B. melitensis from aborted bovine samples. (Hypothesis)
A total of 60 tissue samples (lung, liver and stomach) from aborted bovine fetuses were collected from farms with history of abortion and suspected brucellosis in and around Lahore district. The samples were subjected to AMOS PCR and immunohistochemistry for detection of B. abortus and B. melitensis. Brucella abortus and Brucella mellitensis species specific primer were used to get the desired base pair. The genomic region of B. abortus IS711was amplified at 498bp.
From present study it can be concluded that brucellosis is present in cows and buffaloes at district Lahore and it is more in cattle as compared to buffaloes. Therefore an immediate actions and policies are required to be implemented for the preventing spread of the disease to the other animals and human. For the diagnosis of Brucella species AMOS PCR and immunohistochemistry were used and the results showed that Brucella abortus were more as compared to other species in aborted bovine tissues. The results also showed that the sensitivity and specificity of AMOS PCR is more than immunohistochemistry.
Availability: Items available for loan: UVAS Library [Call number: 2320-T] (1).
27.
Comparitive Immunogenic Evaluation Of Purified And Whole Culture Vaccine Of Pasteurella Multocida B2 Of Bovine Origin
by Anika Khalid (2006-VA-357) | Dr. Imran Altaf | Dr. Jawad Nazir | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pasteurella multocida is responsible for causing Heamorrhagic Septiceimia which is a fatal and acute disease mainly associated with bovines.
Pasteurella multocida consists of two important antigenic structures i-e Outer membrane protein(OMP) and Lipopolysaccharide(LPS) that forms capsule.These two structures played vital role in pathogenicity as well as in the immunogenicity.
In our present project we study the role of LPS,OMP alone and in combination as an immunogens.We compared the immunogenic behaviour of whole culture vaccine(consists of LPS and OMP) purified vaccine(consists of OMP),LPS vaccine (consists of LPS only) by IHA and CFT (using OMP and LPS both as an Ag separately) and the overall results showed that whole culture HS Alum precipitated vaccine produced better antiboby titer against both outer membrane proteins (OMP) and lipopolysacharrides (LPS) when checked against respective antigens as compared to the vaccines containing OMP and LPS alone.
So it was concluded that LPS no doubt act as an immunogen but its immunogenicity increases many times when combine with protein (OMP) as in whole culture vaccine. Availability: Items available for loan: UVAS Library [Call number: 2316-T] (1).
28.
Incidence Of Potential Meat-Borne Pathogens And Their Survival In Ready To Eat Chicken Products
by Sahrish Geaorge (2012-VA-585) | Dr. Jawad Nazir | Dr. Arfan | Dr. Imran Rashid.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: There is an increasing trend of using cooked and semi cooked frozen chicken meat products in Pakistan. These products are susceptible to microbial contamination and often implicated in food borne illnesses. Salmonella, E.coli, Listeria, and Campylobacter are among the major food borne pathogens and are of public health significance which cause thousands of deaths annually. So practice of microbiological safety in production, storage and consumption of these products is of great importance. Microbial contamination of ready to eat chicken meat products is anticipated during processing. Proper cooking and good hygiene practices might be helpful to reduce the microbial load.
Four ready to eat chicken products including shami kebab, sausages, nuggets and kofta kebab of three different companies were purchased from the market after every 15 days interval to have 6 consecutive samples. A total 72 samples were processed and transported to the laboratory at low temperature. The samples were processed to test aerobic plate count, E.coli count and detection of Salmonella. All of the experiments were repeated three times independently. The maximum aerobic count was found in sausages that ranged from to 19.4 × 105 to 1.85×106. The minimum aerobic count was found in semi-cooked nuggets which was 1.3×103. Out of 72 samples, 19 were satisfactory, 35 were acceptable and 18 were unsatisfactory according to microbiological quality standards of ready to eat chicken products.
Availability: Items available for loan: UVAS Library [Call number: 2322-T] (1).
29.
Isolation And Antibiotic Susceptibility Pattern Of Extended Spectrum Β-Lactamases Producing Klebsiella Pneumoniae From Human Clinical Specimen
by Muhammad Tahir Ishaq (2013-va-443) | Dr. Jawad Nazir | Dr. Muhammad Zubair Shabbir | Dr. Aqeel Javeed.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Klebsiella species particularly K. pneumoniae are important opportunistic nosocomial pathogens causing a variety of infections including urinary tract infections (UTIs), pneumonia, septicemia and wound infections. Epidemic and endemic nosocomial infections caused by K. pneumonia species are leading causes of morbidity and mortality throughout the world. Irrational use of antibiotics is creating antibiotic resistance problems in Pakistan. Excessive use of β-lactam antibiotics is responsible for production of ESBL enzymes by Gram negative bacteria especially K. pneumoniae.
A total of 150 samples including urine, pus and sputum were processed for the isolation and evaluation of the ESBL enzyme producing K. pneumoniae as well as their antimicrobial sensitivity pattern. Samples were cultured on cystine lactose electrolyte deficient (CLED), blood, chocolate and McConky agar. Isolates were identified by culture characters, staining reaction followed by biochemical testing through API-20E index system. ESBL production potential was assessed by double disc diffusion method. Out of total 50 urine samples, K. pneumoniae was isolated from 20 samples. Among these 20 isolates, 6 were confirmed as ESBL producers. However, within 50 sputum and pus samples, 13 and 15 isolations were possible out of which 5 and 6 were positive for ESBL production, respectively. Frequency distribution for isolation of K. pneumoniae from urine, pus and sputum samples did not significantly vary from each other. Similarly, ESBL producing potential of all K. pneumoniae isolates also did not significantly vary as p values are higher than 0.05.
A total of 17 ESBL producing K.pneumoniae isolates were tested for their antibiotic susceptibility pattern against 12 commonly used antibiotics i.e. Amoxacillin, Ampicillin, Aztreonam, Ceftazidime, Cefatoxime, Ceftriaxone, Cefixime, Imipenem, Meropenem, Ciprofloxacin, Gentamycin and Amikacin. All of the tested K. pneumoniae isolates showed 100 % sensitivity against amikacin, imipenem and meropenem while found to be completely resistant against rest of the antibiotics.
Molecular confirmation of ESBL production was done through PCR. DNA samples were extracted from ESBL producing isolates. Amplification of the plasmid DNA region (TEM gene) from the DNA samples was done through PCR. The resulting PCR product was run on 1% agarose gel through horizontal gel electrophoresis. Three samples from urine (S1, S4 and S5) and two samples from sputum (S7 and S8) produced required bands while none of the other samples produced any band.
High proportion of ESBL producing K. pneumoniae isolates in present study is an alarming scenario. These organisms are resistant to conventional antibiotics and might spread in the environments thus pose serious threat to the health of human and animal population. Only clinical specimen submitted to the diagnostic lab were tested in present study. Further extensive studies are needed to bridge the gaps in knowledge of antibiotic resistance pattern of K. pneumoniae. Availability: Items available for loan: UVAS Library [Call number: 2366-T] (1).
30.
Effect Of Timing Of Artificial Insemination In Relation To Ovulation On Pregnancy Rate In Sahiwal Cows
by Waqas Ahmad (2007-VA-99) | Prof. Dr. Nasim Ahmed | Prof. Dr. Manzoor Ahmad | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Sahiwal cow is the best milch breed among all Bos indicus throughout the Subcontinent. It is famous for heat resistance, tick tolerance, high milk production. It is also exported to different countries for its peculiar properties. These cows have low fertility when inseminated with frozen semen. Very little fragmented experimental data is available on reproduction of Sahiwal breed, which might be major reason of bad performance from reproduction aspect. The same rule of insemination as we know in Holstein (Bos taurus) when applied on our local animals did not yield similar results. The designed experiment was to determine appropriate time of insemination to improve reproductive efficiency in Sahiwal cows.
This experiment was conducted at Livestock Experimental Station Jahangirabad, District Khanewal Punjab. Eighty five (n=85) adult, multiparous, lactating Sahiwal cows having BCS ≥ 2.5 and weight 325–450 kg, more than 60 days post-partum with normal reproductive tract were selected for this study. Estrous detection was carried out twice daily with teaser bull for 30-45 minutes. Standing heat was considered when cow did not move away for 4-6 seconds with teaser bull being mounted on her. Cows were assigned randomly into four groups (0 h, 12 h, 24 h and 36 h) with respect to standing heat. Frozen semen from bull whose fertility is known to us having post thaw motility of 40% at least was used for insemination.
Ultrasound was used as tool for precise assessment of reproductive status of experimental animals. At the start of each replicate, both ovaries and uterus of all the adult Sahiwal cows were scanned with B-Mode Ultrasound console for presence or absence of fetus and CL or any structural abnormality, with help of 7.5MHz Trans-rectal probe. Pregnancy was diagnosed 35 day post AI. Results were analyzed by using statistical software (SPSS). Pregnancy per AI was compared amongst
CHAPTER 6
SUMMARY
Summary
37
different insemination groups by using binary logistic regression test. Pregnancy was assessed retrospectively by plotting scatter graph using ovulation as our reference point.
The timing of ovulation was 20.64 hours from onset of standing heat. Mean size of ovulatory follicle at 0, 12, and 24 hour after standing heat was 13.52 mm, 14.52 mm and 15.39 mm respectively. The ovulation rate was 97%. Highest pregnancy per AI 57% (13/23) was observed in 0 h group, followed by 36% (8/22) in 12 h, followed by 25% (5/20) on 24 h group. Lowest pregnancy per AI 10% (2/20) was seen in group inseminated 36 hour after onset of standing heat (36 h). While retrospectively highest pregnancy per AI 67% (10/15) was observed in -18 h group, followed by 29% (6/21) in -6 h group and 32% (7/22) in +6 h group, while no pregnancy were observed 0% (0/12) in +18 h group. The overall pregnancy per AI was 33% (28/85) in Sahiwal cows. Availability: Items available for loan: UVAS Library [Call number: 2468-T] (1).
31.
Characterization And Thermostability Of Phytase Produced By Indigenous Aspergillus Niger Isolates
by Madeeha Tariq (2010-VA-293) | Prof. Dr. Aftab Ahmad Anjum | Dr. Jawad Nazir | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Phytase enzyme now becomes more important commercially. Presence of phytate in food
and feed make them less nutritive due to phytate complexes mainly with mineral ions and
proteins. Phytase in monogastric animals and human stomach either produced in small amount or
not. This leads to phosphorous Pi deficiency. Supplementation of food and feed with phytase
enzyme full fill this deficiency through degradation of phytate complexes and release of Pi.
Degradation of phytate complexes makes phosphorous other mineral ions and amino acids
available for growth and development. It was proved that feed conversion rate in poultry
increased due to supplementation of phytase in poultry feed. Feed of monogastric animals mostly
at industrial level pelleted to give it a shape or to kill microorganisms (sterility). At industrial
level enzyme production and processing cost about 2 billion. So this demands a thermostable
phytase to use at industrial level or its cost effective production.
Aspergillus niger have been used industrially for production of beneficial enzymes. A.
niger isolates procured from department of microbiology were confirmed through macro and
microscopic characteristics as A. niger. These isolate were screened for phytase production on
phytase screening medium PSM agar. Positive isolates identified through noval staining using
2% cobalt chloride, 6.25% ammonium molybdate and 0.42% ammonium vanadate for contrast.
Positive isolates next proceeded for phytase enzyme production in broth media (pH 5.6) using
0.5% sodium phytate as substrate. Incubation was done at 30oC for 5-7 days in shaking incubator
150rpm. After production quantification of enzyme was carried out through enzyme activity
assay. There maximum (274.99±10.14 FTU/ml) and minimum (68.88±2.55 FTU/mL) activity of
phytases from isolate PASN01 and PASN08 was observed. Phytases characterized through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) to know protein
molecular weights. Highest molecular weight 107.82kDa was PASN06 and lowest was 35.21kDa
of PASN 01.
Aspergillus niger spores subjected to steam heat treatment at 30oC, 45oC, 60oC, 75oC and
90oC for 15, 30, 45, 60minutes to identify thermostability. At 30oC and 45oC temperature, spores
of A. niger isolates found to be thermostable. But at 60oC, 75oC, or 90oC treatment spores
become inactivated or there 6.0 logarithmic reduction in spore count was observed.
Thermostability of phytases was found at 60oC, 75oC, 90oC for 15, 30, 45, and 60
minutes treatments. Enzyme from A. niger PASN01 and PASN08 observed as thermostable at
60, 75 and 90oC. Phytases from PASN01 and PASN08 showed 160.55±42.96 and 00±.00
FTU/mL decreased in activity after 45 minutes of treatment at 60oC temperature, respectively.
PASN01 phytase displayed 163.88±23.35, 172.77±7.52 and 171.66±7.26 FTU/mL decreased in
activity after 60minutes treatment at 60, 75 and 90oC. In case of PASN08 phytase at 60, 75 and
90oC temperature after 60minutes treatment, 13.33±10.41, 16.66±6.00 and 23.88±41.37 FTU/mL
decreased in activities were observed, respectively. PASN08 phytase observed more
thermostable than other phytases of A. niger isolates. Enzyme can bear pelleting and pre
pelleting temperatures. Enzyme from PASN08 also observed stable during storage at room
temperature.
Conclusion:
A. niger PASN08 spores inactivated or killed and phytase observed stable at 60oC
temperature, after 60mins treatment. Temperature 60oC may be used industrially for cost
effective thermostable phytase production from indigenous A. niger isolate PASN08. Availability: Items available for loan: UVAS Library [Call number: 2475-T] (1).
32.
Characterization And Phylogenetic Analysis Of Hemagglutinin Gene Of Avian Influenza Virus Subtype H9n2 Isolated In 2015
by Arslan Mehboob (2009-VA-76) | Prof. Dr. Tahir Yaqub | Dr. Jawad Nazir | Dr. Maryam Javed.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: H9N2 Avian influenza outbreaks has caused great economic losses to poultry industry resulting
in decrease egg production, high morbidity and mortality. Due to different antigenic variants H9
has become problematic. It has the ability to cross species barrier and increase in pathogenicity.
Hemagglutination inhibition (HI) test is employed extensively for subtyping and detection of
antibody titre against the virus. Continuous mutations in the HA gene transforms AIV subtype
H9N2 into more pathogenic virus that may have pandemic potential and can cross species
barrier. Thus, it was necessary to identify various antigenic variants of H9 virus. It was important to
study the HA gene as it plays vital role in viral attachment, release of genetic material and
pathogenicity.
In present study, a sum of four H9 virus samples were isolated. Both serological and
molecular confirmation was done. 200 samples from different areas were collected and properly
labelled. They were then processed for egg inoculation in embryonated eggs. Virus was grown in
embryonated eggs and harvested fluid is then proceeded for confirmatory testing.
Haemagllutination and Haemagllutination Inhibition testing was done. RNA was extracted by
Kit method and cDNA was synthesized. Reverse Transcriptase (RT-PCR) was performed using
specific primer sets and then the amplicon were run on agarose gel. The bands obtained was sent
for sequencing and Phylogenetic analysis was obtained using software and tree was constructed.
Protein analysis was also performed.
The present study enabled us to characterize and construct Phylogenetic tree of HA gene of
currently prevailing H9N2 Avian Influenza isolates in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2474-T] (1).
33.
Comparative Potency Of Two Different Trivalent Vaccines Against Foot And Mouth Disease In Cattle Around The Area Of Ravi Campus Pattoki
by Muhammad Fahimullah Khan (2009-VA-137) | Prof. Dr. Aneela Zameer Durrani | Dr. Muhammad Ijaz | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Foot and Mouth Disease is a highly contagious viral disease of all cloven footed animals.
The best control strategy of this disease is effective and in time vaccination. A successful
vaccination campaign depends on the serotype identification and specific vaccination against the
prevalent serotype of the virus. The present study was designed to evaluate comparative potency
of two anti-FMDV vaccines (UVAS-FMD, Deccivac Intervet) used in cattle around areas of
Lahore Pakistan. Blood samples were taken from vaccinated animals on day 0, 30, 60 and 90
post priming. Antibody titer was evaluated with different route of administration and various
adjuvant based vaccines. Four animal groups were made each containing 5 animals; in group 1
UVAS vaccine was used by Sub/ Cut route (gel based) at priming dose, followed by Intra
Muscular (oil) on 30th day. In group 2 UVAS vaccine was given I/M (oil) as priming dose and
booster (oil) I/M. In group 3 Deccivac (oil) vaccine was used I/M for priming and boosting. In
group 4 Deccivac (oil) vaccine was used as Sub/ Cut for priming and boosting. The results
revealed non-significant difference (p>0.05) among the four different groups administered with
FMDV vaccines when evaluated at day 30 and significant difference (p<0.05) at day 60 and 90
post vaccination. Analysis of variance showed significant difference (p<0.05) in antibodies
between groups and with in groups at day 60 and 90. Gel based vaccine gave quick antibody
response which later maintained with oil based booster dose. The difference in antibody titers
obtained in the present study was found non-significant (P>0.05) between the antibody titers of
FMD trivalent vaccine of UVAS and Deccivac at 90th day of inoculation. There was significant
difference (p<0.05) between the adjuvants of vaccine. Animals inoculated with priming dose of
gel based vaccine followed by oil based boosting showed significantly high anti FMD antibody
titer than animals inoculated with oil for both priming and boosting. There was significant
Summary
54
difference (p<0.05) between the groups vaccinated with various routes of administration. The
animals inoculated with priming dose through s/c followed by boosting dose i/m showed
significantly high anti FMD antibody titer at 90th day of inoculation compared with those
inoculated intramuscularly for both priming and boosting. The animals inoculated with oil based
vaccine for both priming and boosting through S/c showed marked significant decreased in anti
FMD antibody titer. The route of administration revealed significant difference (p<0.05) in
antibody response within groups and between groups at day 30, 60 and 90. In all three readings
the mean for sub/cut priming and IM boosting were found significantly high (p<0.05) as
compared to other routes. In conclusion it is recommend from the study that FMD vaccination
with sub/cut priming and booster dose with IM route. Availability: Items available for loan: UVAS Library [Call number: 2499-T] (1).
34.
Effect Of Synchronization Protocols (Pg And Ovsynch) On Estrus Response, Estrus Intensity, Ovulation Time And Conception Rate In Cholistani Cows
by Muhammad Awais Ajmal (2009-VA-370) | Dr. Aijaz Ali Channa | Prof. Dr. Nasim Ahmad | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Cholistani cattle breed, like other milch breeds of our country (Sahiwal, Red Sindhi cattle), is one of the indigenous breeds of Pakistan having superior dairy characteristics. Being heat tolerant breed present in tropical and subtropical areas of Pakistan it has average milk production 1235 litters with 4.8% fat in it. To minimize the cross breeding of Cholistani cows, some efforts are being done for its conservation, through management and genetic improvement. Moreover genetic improvement, through artificial insemination (A.I) is in process. In recent times another important tool Fixed time artificial insemination (FTAI) has emerged to improve fertility which usually is performed in association with a variety of estrus and ovulation synchronization regimens. Synchronization of estrus is actually to bring large group of females in estrus at a desired fixed time by manipulation of estrus cycle. Two synchronization protocols were compared. In PG group, each animal was treated with luteolytic dose of PGF2α (d-Cloprostenol 0.150 mg; Dalmazine, Fatro®, Ozzano Emilia Italy; 2 ml; i.m) on random stage of the estrous cycle and repeated after 11 days TAI was done at 72 and 84 h after 2nd PG. In OVS group each cow received an intramuscular injection of gonadotropin releasing hormone (GnRH; 50 mcg of a GnRH analogue, Dalmarelin TM Fatro®, Italy; 2 ml; i.m) on random stage of estrus cycle (day 0). On day 7 these cows were treated with PGF2α (d-cloprostenol 0.150 mg; Dalmazine, Fatro®, Ozzano Emilia Italy; 2 ml; i.m), followed by second injection of GnRH on day 9 and TAI was done at 12 and 24h after 2nd GnRH. Estrus response and estrus intensity were higher in OVS group as compared to PG. Timing of ovulation was shorter in OVS group as compared to. Conception rate are also high in OVS group. Therefore it is concluded that Ovsynch protocol is helpful to improve ovulation and conception rate in Cholistani cows. Availability: Items available for loan: UVAS Library [Call number: 2523-T] (1).
35.
Isolation And Molecular Characterization Of Rotavirus From Calf Diarrhea And Preparation Of Vaccine
by Nadia Mukhtar (2008-VA-718) | Prof. Dr. Tahir Yaqub | Dr. Jawad Nazir | Prof. Dr. Asim Aslam.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The main contribution of the thesis “Title” is threefold. First, rotavirus was isolated and identified from calf diarrhea samples from 10 districts in Punjab. Second, optimization of molecular diagnostics and genome sequencing was done of the positive bovine rotavirus isolates from Pakistan. And thirdly, the preparation as well as evaluation of killed vaccine against bovine rotavirus isolates was performed.
The above three objectives of this study were created due to the distribution of rotavirus all over the world as an enteric pathogen in both human as well as animal species. In developing countries where cases of malnutrition are very common in young children and animals, this virus has a special importance as an etiologic agent. It causes severe diarrhea, when accompanied with severe dehydration, leads to high rate of mortality. Among the rest of the infectious diseases present in calves, neonatal diarrhea is a dire threat as it has a major impact on economic viability. Calf diarrhea is the most important problem in dairy calves that causes more financial losses to the calf producers than any other. Although numerous etiological agents may be implicated, Rotaviral diarrhea is one of the main infections causing calves to scour between five to fourteen days of age.
The cattle and buffalo calves’ population in Pakistan is devastatingly affected by the neonatal calf diarrhea due to rotavirus outbreaks. Neonatal calf mortality varies from 8.7 to 64 per cent throughout the world accounting for 84 per cent of the total mortality in the first month of age and is particularly high in the third week. While vaccination is available for the disease, it is being imported in Pakistan from other countries. The importation of the said vaccine thus, leads
SUMMARY
117
to extra expenses for the farm managers. As mentioned above one of the aims of this study is to develop an effective vaccine against bovine rotavirus and cut down expenses for farm managers.
To fulfill the objectives proposed in this thesis, rectal swabs and fecal samples were collected from public/private sector buffalo and cattle farms from 10 districts of the Punjab: Lahore, Faisalabad, Okara, Sahiwal, Sargodha, Chakwal, Bhakkar, Bahawalnagar, Multan and Bahawalpur. The samples were selected on the basis of agro-ecological zones of the province. As sampling based on agro-ecological zones allow for better data collection for recording incidence rate of the disease. Samples (n=10) from each diarrheic and apparently healthy cattle and buffalo calves from all of the districts were collected. In this way a total of 200 samples from buffalo calves and 200 samples from cattle calves were collected for this study.
Antigen of bovine rotavirus was screened from calf feces through Direct Sandwich ELISA. Bovine rotavirus samples were further confirmed through the amplification of the VP4 and VP6 genes through Rt-PCR. Homology and phylogenetic analysis of the sequenced samples was also performed. The data gathered through this analysis was helpful in collecting important data regarding the similarities as well as differences of the bovine rotavirus strain present in Pakistani isolates when compared to local regions as well as international ones. The data is also valuable when it comes to production of effective vaccines again rotavirus. RNA viruses are known to mutate unpredictably and it is safe to assume that a particular vaccine might not work effectively against all strains of a particular virus. That’s why analysis of data pertaining to all possible BRV strains is important for creation of an effective vaccine of import quality in order to help the economy of Pakistan.
Rotavirus isolate, after adaptation on MDBK cell line, was further propagated to determine TCID50 for vaccine preparation purposes. Final dose of the vaccine was adjusted to
SUMMARY
118
approximately 3ml, containing 40% culture and 60% adjuvant. Final vaccine contained 1ml of inactivated bovine rotavirus harvested culture, 1.8ml of Montanide ISA 70, 0.2ml of PBS and 0.05% of Thiomersal sodium. Efficacy of the vaccine was checked in rabbits.
For vaccine efficacy testing twenty one month old rabbits were procured. Rabbits were reared in individual isolator units in the shed facility of Quality Operations Laboratory, UVAS, Lahore. The collected rabbits were divided into two groups, vaccinated and unvaccinated rabbit groups. Each group had 10 rabbits. One ml of rotavirus vaccine was administered intramuscularly in vaccinated rabbits group. In unvaccinated rabbits group 1ml of normal saline was injected intramuscularly. The second dose of vaccine was administered at 24 days post-vaccination of first dose. The rabbits from both groups were bled at 0, 14, 28 and 42 days post-vaccination. The antibody response of rabbits to rotavirus vaccine was determined through using Antibody detection kit. The rabbits were challenged on day 42 post-vaccination using live field strain of rotavirus having TCID50 1 × 108.5. The rabbits were observed daily up to 14 days post-vaccination for appearance of diarrheic signs. The stool samples of ELISA positive were further confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR) at least 14 days post-vaccination.
The field trials were conducted at Livestock Production Research Institute, (LPRI) Bahadurnagar, Okara. The field study was done to evaluate the prepared rotavirus vaccine for prevention of neonatal calf diarrhea. For this trial, 100 dams were selected. The dams were divided into two groups and each group consisted of 25 pregnant cows and 25 pregnant buffalos. A total of 50 dams (25 cattle and 25 buffalo) were vaccinated intramuscularly with 3ml of prepared inactivated rotavirus vaccine. The 50 remaining dams (25 cattle and 25 buffalo) were kept unvaccinated.
SUMMARY
119
The blood samples were collected for serum separation after 0, 14, 28 and 42 days post vaccination in dams. The antibody titers were measured using antibody detection ELSIA kit.
After calving, newborn calves were fed with the colostrum obtained from the vaccinated dams daily for 5 consecutive days. Similarly, the calves from unvaccinated dams were fed on colostrum from their unvaccinated dams.
The 5 calves from vaccinated and 5 from the unvaccinated dams were isolated in individual isolators. These calves were challenged orally with 1ml of live field strain of rotavirus having 1 × 108.5 TCID50 and the animals were observed for diarrheic signs for 7 days.
All of the collected data was subjected to statistical analysis of (one way) ANOVA and t-test using SPSS. The <0.05 p-value determined the significance of the results through this study.
The data collected through this study allowed for the creation of valuable inferences. According to the current results of this study, the prevalence of bovine rotavirus was shown to be 6% in Punjab. This 6% included 40% and 20% from the districts of Lahore and Faisalabad respectively. Keeping these results in mind, it is to be noted that the recorded prevalence percentage from this study is higher than the prevalence of 2% in Lahore according to a previous study done in the country. It is to be noted that while the 6% prevalence of rotavirus in Punjab detected through ELISA is lower than the prevalence of 16.83% which was detected by ELISA in diarrheic calves from pervious researches, the 12% prevalence detected by ELISA in this research is higher than the prevalence of 7.25% detected by ELISA in diarrheic calves from past data.
In the present study of this thesis it was observed that the use of killed vaccine for bovines produced more efficient immune response in calves. It also enhanced the clostral rotavirus antibody titers as compared to previous studies where the use of the same strain of modified-live virus in a commercial vaccine administered IM with or without adjuvant did not significantly
SUMMARY
120
elevate colostrum antibody titers. The results collected from the present research showed that the average antibody titers in the 25 cattle dams at 0, 14, 28 and 42 days post vaccination were 0%, 57%, 68% and 78% respectively. In a similar manner the average antibody titers in the 25 buffalo dams at 0, 14, 28 and 42 days post vaccination were 0%, 55%, 70% and 82% respectively. These results indicated
the protective maternal antibody level against the rotavirus which will be transferred passively to calves. The results indicate that vaccinated dams were able to provide passive immunity to both buffalo and cattle calves in order to provide protection against the deadly virus. Availability: Items available for loan: UVAS Library [Call number: 2570-T] (1).
36.
Monitoring Of Humoral Immune Response Of Monovalent And Combined Ppr And Fmd Serotype “O” Virus Vaccine In Small Ruminants
by Mudassar Hameed (2009-VA-386) | Dr. Jawad Nazir | DR. M. ZUBAIR SHABBIR | DR. MUHAMMAD IMRAN.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: PPR is an acute and highly contagious viral disease of small ruminants caused by morbilivirus. It causes high morbidity and mortality in small ruminants and heavy economical loses to farmers. Live attenuated vaccines are commonly used to control PPR. FMD is another highly contagious viral disease of cloven hoofed animals caused by picorna virus. Its severity is relatively high in large ruminants but carrier status of small ruminants is usually observed. In large ruminants killed FMD virus vaccines are routinely used but in small ruminants it is not practiced in Pakistan. There was need to formulate a combined vaccine containing both PPR and FMD viruses which will help to control both of the diseases in small ruminants.
.
A total of 100 goats were divided into 10 groups comprising 10 animals in each group. Each of the vaccine such as PPRV, FMDV and PPRV+FMDV was be prepared without adjuvant, gel and oil based. A total of nine types of vaccines were inoculated in the respective groups while one group remained un-inoculated negative control. Each group was subdivided into two subgroups (n=5). One subgroup was received single dose and the other inoculated with two doses of the vaccine. Serum samples from each goat were collected at 0, 1, 2, 3, 4, 5, and 6 months post vaccination (PV) and kept frozen at -20 ºC. Immune response of the vaccinated animals was monitored by measuring antibodies against PPR and FMD viruses through cELISA and VNT.
Results of the present study showed that mean percentage inhibition (MPI) value against PPR virus of non-adjuvant, gel and oil based combined (PPR+FMD) vaccines at six month post-vaccination was 83.46 ± 2.25, 80.27 ± 2.13 and 82.16 ± 1.70 respectively, whereas mean
x
neutralization antibody titer (MNA) was 4.39± 0.37, 4.06± 0.26 and 4.49 ±0.46 respectively. MPI value against FMD virus of combined (PPR+FMD) non-adjuvant, gel and oil based vaccines at six month post-vaccination was 90.17 ± 1.15, 67.22 ± 3.14 and 72.22 ± 2.04 respectively, whereas mean neutralization antibody titer (MNA) was 2.33 ± 0.27, 1.47 ± 0.10 and 1.83 ± 0.16 respectively. These MPI and MNA values showed that immune response against PPRV of combined vaccines was equivalent but non-adjuvant combined vaccine have evoked higher titer followed by oil and gel based vaccines against FMDV.
MPI values of non-adjuvant, gel and oil based monovalent PPRV vaccines at six month post-vaccination was 81.46 ± 2.22, 80.12 ± 2.13 and 81.28 ± 0.70 respectively, whereas mean neutralization antibody titer (MNA) was 4.59 ± 0.17, 4.25± 0.06 and 4.51 ±0.12 respectively.
MPI values of non-adjuvant, gel and oil based monovalent FMDV vaccines at six month post-vaccination was 00.00 ± 0.00, 82.23 ± 4.18 and 90.22 ± 0.43 respectively, whereas mean neutralization antibody titer (MNA) was 0.00 ± 0.00, 1.63 ± 0.10 and 2.99 ±0.16 respectively.
These MPI and MNA values showed that monovalent PPR vaccines induced equivalent immune response in all three formulations but monovalent FMD vaccines MPI and MNA values showed that oil based vaccine has provoked significantly higher titer followed by gel based vaccine. Whereas non-adjuvant FMD vaccine titer was diminished at one month post vaccination. Booster vaccine shots provoked higher antibody titer than single shots in all various formulations of vaccines.
The data thus obtained was analyzed through One Way ANOVA followed by Randomized Complete Block Design (RCBD). Availability: Items available for loan: UVAS Library [Call number: 2635-T] (1).
37.
Effects Of Physico-Chemical Properties Of Diluents On The Infectivity Titers Of Freez Dried Ppr Virus Vaccine
by Fariya Yaqub Baig (2009-VA-240) | Dr. Jawad Nazir | Prof Dr. Aftab Ahmad Anjum | Dr. Waseem Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: 6.1. Introduction.
Peste des Petits Ruminants (PPR) is a febrile viral disease of sheep and goats. The disease is
responsible for low productivity in small ruminants and great economic losses to the farmers in
Africa and Asia including Pakistan. There is no specific treatment for PPR and prevention is only
possible through the use of live attenuated vaccines. Being an enveloped virus, PPRV is heat
sensitive. Poor immunological responses have been observed after the vaccination of PPR.
Reasons may be disturbance in maintenance of cold chain, improper handling and route of
vaccination as well as reconstitution in inappropriate diluents. Physiochemical properties
(temperature, pH, osmotic pressure, salinity and UV light) of diluents effects the infectivity of
PPR freeze dried vaccine as live virus vaccines work properly after reconstitution within the
recommended time interval.
6.2. Experimental Design.
Effect of three diluents (normal saline, phosphate buffer saline and distilled water)
adjusted to various pH conditions (5.00, 6.00. 7.00, 8.00, and 9.00) on the infectivity of PPRV
was evaluated. Contents of the freeze dried PPR vaccine vials were diluted with one ml of the
respective diluent adjusted to above mentioned pH conditions. Virus infectivity from the vials
was measured immediately following reconstitution. One set of vials was kept at room
temperature (25 ºC ±2) and virus infectivity was measured afterwards at 30, 60, and 120 minutes
as described in the section 3.3.2. While other set of the vials was kept at refrigerated temperature
(4 ºC ±2) and virus infectivity was measured at 30, 60, 120, and 180 minutes as described in the
Summary
46
section 3.3.2. The change in pH of each vial following reconstitution was also measured
accordingly. Each set of experiment was repeated three times independently.
6.3. Results.
PBS and NS gave equivalent results and proved better than distilled water to restore the
infectivity of PPRV. For a better comparison of virus stability in the PPR freeze dried vaccine
following reconstitution in various diluents adjusted to various pH conditions the infectivity titer
of the virus in the vaccines were measured at various time points. The serial data thus obtained
was analyzed by linear regression model to calculate T-90 values (Time required for 90 %
reduction in virus infectivity). The higher T-90 values indicate better stability of the virus at a
designated condition. At both of the temperatures the T-90 values were equivalent and relatively
higher for all of the diluents adjusted to pH 7.00 and 8.00 while these values are lower at extreme
pH conditions. Minimum T-90 values (126± 56) were observed at pH 9.00 in normal saline kept
at ambient temperature while maximum T-90 (448 ± 49.7) values were observed at pH 7.00 in
phosphate buffer saline kept at refrigerated temperature.
6.4. Conclusion.
Results of the present study suggest that virus infectivity in the live attenuated PPRV vaccine can
be better stabilized following reconstitution in PBS adjusted to neutral or slightly alkaline pH.
Chilled vaccine diluent is preferable to the one kept at room temperature and vaccine should be
administered within 30 minutes of reconstitution. Availability: Items available for loan: UVAS Library [Call number: 2645-T] (1).
38.
Effect Of Colchicine On Cellular And Humoral Immune Responses In Mice
by Shahzada Khurram Syed (2007-VA-444) | Dr. Aqeel Javeed | Prof. Dr. Muhammad Ashraf | Dr. Jawad Nazir | Dr. Shahbaz Yousaf.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Colchicine is a medication that treats gout. It is a natural product and secondary metabolite, originally extracted from plants Colchicum autumnale .It causes modulation of chemokine and prostanoid production and inhibition of neutrophil and endothelial cell adhesion molecules by which it interferes with the initiation and amplification of the joint inflammation.
The present study is designed to evaluate the effects of colchicine on cellular and humoral immunity in mice. There were five groups for each assay i.e. group I (negative control), positive control and three colchicine treated group II (40μg/kg), group III (80μg/kg) and group IV (160μg/kg). The number of mice in each group was five to eight. All these groups were administered doses intraperitoneally. To determine the effect of colchicine on cell mediated immunity , delayed type hypersensitivity (DTH) assay, macrophage engulfment assay, cyclophosphamide induced neutropenic test and nitric oxide production was performed .DTH was performed by measuring skin thickness. DTH showed significant difference (P<0.001) of negative control to colchicine treated groups 40μg/kg, 80μg/kg and 160μg/kg. With increasing dose, there was decrease in skin thickness of the mice. Highest reduction of skin was found at 160μg/kg. Macrophage engulfment assay was performed to evaluate the effect of macrophage induced phagocytosis. There was significant ( P <0.001) difference of engulfment of SRBCs by macrophages with negative control to colchicine treated group II (40μg/kg), group III(80μg/kg) and group IV(160μg/kg) groups. There was significant difference of engulfment of macrophages at 45 and 90 minutes.
Cyclophosphamide induced neutropenic test was performed to assess the effect of colchicine on total leukocyte count (TLC) and differential leukocyte count (DLC). There was
SUMMARY
77
reduction of TLC to about 45.3% in control to 48.3%, 54.68% and 65.42% in group II (40μg/kg), group III (80 μg/ kg) and group IV (160μg/kg) respectively when these were compared with primary values of TLC. There was significant difference of reduction in the neutrophil count of negative control 1057 (±120) to 902 (±67) in group II (40μg/kg), 734(±69) in group III (80 μg/ kg) and 609 (±71) in group IV (160μg/kg) of doses of colchicine. This test showed that with the increasing dose of colchicine, there was significant (P<0.001) difference of TLC count and neutrophil count.
Nitric oxide (NO) production by macrophages was performed for measuring different concentrations of nitric oxide produced. There was significant difference (P<0.001) in NO production by macrophages alone and LPS stimulated between negative control to group II (40 μg /kg), group III (80μg/kg), group IV (160μg/kg) of colchicine. With increasing dose, there was significant reduction in production of NO. There was significant P<0.0001 reduction in body weight andspleen weight difference of mice in different groups of colchicine treated 40μg/kg, 80μg/kg and 160μg/kg from negative control after treatment. There was difference of weight of Thymus of group II (40 μg/kg), group III (80μg/kg) and group IV (160μg/kg) but difference was statistically not significant. There were no histopathological changes observed in spleen and Thymus at 40μg/kg and 80μg/kg doses of colchicine. At 160μg/kg dose, increase in thickness of trabecular was seen .due to edema in the spleen. For evaluation of colchicine effect on humoral immunity, haemagglutination assay, mice lethality test and Jerne hemolytic plaque formation were performed. Haemagglutination assay (HA) was performed by using red blood cells injected intraperitoneally in mice to measure antibody titer. There was significant difference of (P >0.001) to colchicine treated group II (40μg/kg), group III (80μg/kg) and group IV (160μg/kg)with group I (negative control).With the increasing dose, there was reduction in the
SUMMARY
78
HA titer. Mice lethality test was performed by testing immune response of the mice to the challenge infection of P.multocida. It was performed by comparing mortality ratio of mice after administration of drug. There was no death of mice in the negative control group in which there was administration of PBS and vaccine. At 40μg/kg dose of colchicine, there was 50% mortality ratio. At 80μg/kg dose of colchicine 75% mortality ratio was observed. Maximum mortality ratio was observed at the 160μg/kg colchicine dose i.e. 100%.
Jerne plaque formation test was performed and plaques formed was enumerated and recorded as the number of plaque forming cells (PFCs) per million cells. There was significant difference (P<0.001) of reduction in number of plaques from negative control to all doses of colchicine 40 μg/kg, 80 μg/kg and 160μg/kg. Antibody formation was decreased with increasing the dose of colchicine. Therefore, it is concluded that colchicine suppresses the cellular and humoral responses in mice. Availability: Items available for loan: UVAS Library [Call number: 2650-T] (1).
39.
Pathobiological Studies Of Canine Parvo Virus Infection Currently Prevailing In Dogs In Pakistan
by Urooj Fatima (2009-VA-69) | Dr. Muti-Ur-Rehman Khan | Dr. Raheela Akhtar | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Canine Parvo virus infection is causing fatal disease to dogs resulting in severe morbidity and mortality. It is becoming disastrous for canids in Pakistan. It is important to study the pathogenesis of this deadly virus. Parvovirus infection has become more pathogenic with passage of time and its pathogenicity may be enhanced due to possible mutation in field virus. The possible mutations may be the cause of vaccine failure and rapid outbreaks.
In present study, sum of 50 fecal samples from different areas were collected and properly labelled. Both serological and molecular confirmation was done.
Haemagglutination test was used as a screening test. HA test was performed to check agglutinating activity of samples as described by (Desario et al. 2005). Haemagllutination was done and only 20 samples were found positive. Positive samples were selected for polymerase chain reaction. DNA of selected samples was extracted through kit method according to protocol provided by the manufacturer. Nano drop was done to check quantity and purity of viral DNA. PCR was performed. Already reported primers were used for detection of CPV (Buonavoglia et al. 2001). The PCR product was confirmed by running the PCR product on 1.2% agarose gel. Electrophoresis was performed at 110 Volts for 30 minutes. The amplified PCR product was further purified using GeneJet Gel Extraction Kit using instructions provided by the manufacturer. Purified samples were sent to commercial lab for Sanger sequencing. The obtained sequences were examined using BLAST (Basic Local Alignment Search Tool). Sequences were compared with other sequences already obtainable from the GenBank database (http://www.ncbi.nlm.nih.gov) and were carefully aligned by BioEdit software (version 7.2.5.0) using the ClustalW method. The MEGA software program, version 6.0 was used to construct a phylogenetic tree using the neighborhood joining method.
CHAPTER 6
SUMMARY
Summary
40
Complete blood count was performed on whole blood samples. The samples were divided into two groups (infected & non-infected) based on the results of PCR. Paired T-Test was applied on the results obtained and results showed that all parameters statistically showed significant difference between two groups except WBC’s but still the number of WBC’s decreases in infected animal like other parameters. Serum chemistry analysis was performed on serum samples. These samples were also divided into two groups (infected & non-infected) based on the results of PCR. Paired T-Test was applied on the results obtained and results revealed that all parameters statistically showed significant difference between two groups except ALT and creatinine levels.
Results showed that new CPV-2a was found to be the major circulating strain of Pakistan as out of 7 sequenced samples, 6 were identified as CPV-2a and only 1 of them was CPV-2. PCR was found to be the most sensitive assay for the detection of CPV, as compared to HA and ICT- CPV antigen detection kit. The strain in vaccine was identified as CPV-2, this finding was considered important because it indicates one of the reason for vaccine failure. The phylogenetic analysis of strains showed that all the samples collected from different regions of Pakistan falls in one clade and branching pattern is shared with the neighboring countries especially China. This information reveals that our virus is genomically similar to our neighboring countries India and China. Upon hematology and serum chemistry analysis, it was revealed that the virus does effect the blood parameters and all the red blood cell indices were lowered except platelet count.
The present study enabled us to study pathogenesis, to determine currently prevailing strain of CPV in Pakistan, identify the cause of disease occurrence even after vaccine, to study pathogenesis of CPV, to characterize and construct phylogenetic tree of currently prevailing canine parvovirus field isolates. Availability: Items available for loan: UVAS Library [Call number: 2662-T] (1).
40.
A Comparative Epidemiological Study Of Coccidiosis In Broilers Raised Under Open And Control Sheds
by Shehar Yar Alvi (2007-VA-173) | Prof. Dr. Khalid Saeed | Dr. MUhammad Lateef | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The domesticated fowl (Gallus gallus) is susceptible to seven species of genus Eimeria which are are Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox and Eimeria tenella. All of these are capable of causing disease but the clinical picture and pathogenesis may be different according to species, while the pathogenicity ranges from mild to severe. All the species are ubiquitous and cause disease in combination up-to 6 species at the same time on an individual farm so in this sense coccidiosis may be regarded as a disease complex. Now a days subclinical coccidiosis is more frequently affecting the birds as compared to clinical coccidiosis and greatest financial losses are being caused by subclinical coccidiosis in terms of decreased or less weight gain and reduced feed conversion efficiency.
. The present study was designed to compile data on the prevalence of coccidiosis in broilers reared under open and controlled sheds situated in and around the Lahore city. Study provided better understanding of the risk factors associated with coccidiosis and their relationship.
A questionnaire was designed to record information regarding the management practices, health status of the flock, weight gain. Pooled faecal samples were collected from 50 control sheds and 50 open sheds and were transported to the parasitology laboratory of UVAS. Faecal sample were examined by direct smear to see the coccidial oocysts. Post mortem was conducted to check the presence or absence of the gross lesions associated with coccidiosis. Association between coccidiosis and the risk factors was determined, and the results of open and control sheds were compared. It was assumed that coccidial infections will be higher in the open sheds as compared to environmentally controlled sheds. Open sheds had more prevalence 78% as compared to closed sheds which was reported as 72%. Five major risk factors were studied. Temperature and humidity fluctuation were strong risk factors associated with prevalence of coccidiosis. While litter condition also appeared as an associated risk factor for the prevalence and occurrence coccidiosis in both type of farming systems. Whereas use of medicated feed in open houses appeared as an associated risk factor but in controlled houses use of medicated feed was not associated with the prevalence of coccidiosis. History of previous infections of coccidiosis was also associated risk factor in both type of farming systems. The high prevalence of coccidiosis in open sheds may be due to lack of biosecurity and uncontrollable conditions of temperature and humidity while closed farms have proper biosecurity measures and good husbandry practices. Use of medicated feed and good husbandry practices may be help full to minimize the risk of occurrence of coccidiosis. Further studies are required for better understanding of the disease and associated risk factors.
Therefore, the following recommendations are forwarded.
• Educating farmers about the importance coccidiosis and its control.
• Adaptation of good management practices on farms.
• Avoid over-crowding in the house.
• Alternative remedies need to be developed and evaluated to prevent and control coccidiosis.
Availability: Items available for loan: UVAS Library [Call number: 2691-T] (1).
41.
Prevalence Of Cpb2 In Clostridium Perfringens Isolated From Livestock In Punjab Pakistan
by Iqra Baig (2009-VA-228) | Dr. Aamir Ghafoor | Dr. Jawad Nazir | Dr. Waseem Shahzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The current study designed for the microbiological and molecular characterization of
C. perfringens. C. perfringens is Gram positive rod shape, spore forming bacteria and these are free living bacteria. It is suggested by different studies that the C. perfringens has ability to producing different disease in intestine wall of bovines.
Mostly C. perfringens has ability of producing different variety of toxins and enzymes that are responsible for severe myo-necrotic lesions. C. perfringens is divided into five groups (A-E) on the basis of ability for producing the major toxins (alpha, beta, epsilon and iota). A total of 100 fecal samples cattle (n=50) and buffalo (n=50) were collected and analyzed to determine the prevalence of C. perfringens contamination. The samples were enriched in Fluid Thioglycollate Medium (FTM), purified on Reinforced Clostridium Medium (RCM) agar and were identified by their culture characters, morphology and biochemical profile. For confirmation of C. perfringens biochemical test, hemolysis on blood agar, lecithinase activity, gelatine liquefaction test and nitrate reduction test were performed. C. perfringens was successfully isolated from 17 out of 100 samples with an overall positivity ratio of 17 percent. Seven out of 50 were positive for C. perfringens in cattle while this was 10 in buffalo showing slightly higher percentage of C. perfringens in buffalo. Confirmed C. perfringens isolates through biochemical test were subject for DNA extraction by boiling method and kit method both. Isolates of C. perfringens confirmed through biochemical testing were subject for PCR to confirm cpb2 toxin gene. Zero prevalence was found from PCR results.
CHAPTER 6
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47
Conclusion:
The study reveals that there is high prevalence of C. perfringens among buffalo than cattle. C. perfringens is prevalent in our local animals but beta2 toxin gene not found in the C. perfringens. Availability: Items available for loan: UVAS Library [Call number: 2209-T] (1).
42.
Comparative Efficacy Of Water Sanitizers And Ozonisation To Improve Microbiological Quality Of Poultry Drinking Water
by Saher saeed(2011-VA-395) | Dr. Jawad Nazir | Dr. Arfan Ahmad | Dr. Aqeel Javeed.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2017Dissertation note: Water is a vital nutrient and plays significant role in poultry metabolism, digestion and absorption of food. One of the most important segments in health management for poultry production is water quality. Drinking water of poultry may act as source of microbial Mirobiological quality testing of the water is necessary for human and animal consumption. Acceptable limit of fecal coliform and total coliform for poultry drinking water is zero and 50 CFU/ml, respectively. This is why proper treatment of the water to reduce bacterial loads is highly recommended.
The antimicrobial efficacy of four water sanitizers and ozone was tested and compared in reducing the microbial counts in artificially contaminated water. Water sample collected from a commercial poultry farm was artificially contaminated with ATCC culture of E. coli (1.0 McFarland units). The water sample was treated with sanitizers and ozone at recommended dose and contact time period. After each experiment, microorganisms were recovered and enumerated by spread plate method. For each disinfectant, residual antimicrobial activity was also checked at regular intervals of one hour up-to four hours post treatment. Each experiment was performed in triplicate. Efficacy of disinfectants was measured as log reduction values were calculated after enumeration of microbes on treated samples and untreated samples. The results were analyzed by one way ANOVA using SPSS software.
All of the sanitizers and ozone treatment at recommended doses resulted into more than two logs reduction in the microbial counts. Ozone treatment of the water samples resulted into maximum log reduction following initial interaction. Mean log reduction values (MLR) for ozone at 15, 60, 120, 180 and 240 minutes post treatment are 2.65, 3.74, 3.64, 4.44 and 5.40 respectively.
Summary
60
Statistical analysis show that the MLR within all sanitizers and ozone did not significantly vary from each other at 15 minutes, one hours and three hours post treatment. At 2 hours post treatment MLR value of Quatovet was significantly higher as compared to other sanitizers and ozone. While at four hours post treatment Dutrion and Quatovet treated groups have significantly higher log reduction values in comparison to other sanitizers and ozone
Results of present study show that all of the tested water sanitizers and ozone can destroy more than 99 % of the microbes present in the water after treatment with the recommended doses. Ozone has the highest efficacy among all sanitizers following initial treatment. However, QAC based (Quatovet) and chlorine based (Dutrion) sanitizers have maximum residual antimicrobial activity. Keeping in view of the efficacy and safety of the tested products, the QAC are supposed to be superior among all other agents. Availability: Items available for loan: UVAS Library [Call number: 2828-T] (1).
43.
Antiviral Potential Of Gold And Silver Nanoparticles Against Newcastle Disease Virus In-Vitro
by Anam Iftikhar (2011-VA-404) | Dr. Jawad Nazir | Dr. Muhammad Nawaz | Dr. Aqeel Javeed.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Nanotechnology supplies a golden platform, where properties of pure metals are modified and improved by converting them into their nanoparticles, and it is applicable to numerous fields such as diagnostics, antimicrobial agents and drug delivery. Nanoparticles act as bactericidal, antifungal and antiviral agents. Several types of nanoparticles (gold,copper,silver,aluminium,magnesium,zinc and titanium) have been reported in literature out of which are known to have antibacterial properties. Viral diseases present challenging problems worldwide. Newcastle disease (ND) is endemic in Pakistan, and is linked with huge economic disastrous to farmers. Application of antiviral therapy to control the active infection of NDV is very limited. Use of nanotechnology to control active virus infection might be a viable solution to limit the disease in effected flocks.
In this study four groups of nanoparticles i.e. gold, silver, magnetic and gold coated magnetic were evaluated against mesogenic strain of Newcastle disease virus. Chicken embryos were used to propagate the virus and infective amniotic allantoic fluid was collected. Evaluation of nanoparticles in reducing virus infectivity as a measure of tissue culture infective dose (TCID50) was performed. Various dosesof nanoparticles (very low, low, medium, high and very high) were allowed to interact with virus suspension in three ways i.e.pre- treatment, post-treatment and co-treatment methods. Virus infectivity before and after the treatment with nanoparticles was measured and subsequently used to calculate reduction factor (RF). All of the experiment was repeated three times.
It was observed that in pre and post-treatment, silver, gold coated magnetic and magnetic nanoparticles groups the infectivity titerswere efficiently reduced at high dose. While in co-treatment, silver, gold coated magnetic and magnetic nanoparticles groups the virus inactivation rates were relatively higher at low and very low doses.
It is evident from the findings that within the tested nanoparticles, silver, coated magnetic and magnetic nanoparticles have equivalent antiviral properties against NDV. While within various treatment methods co-treatment assay proved to be more effective in reducing virus infectivity than the pre and post treatment group. The results of present study are suggestive of testing antiviral properties of the nanoparticles in vivo conditions.
Availability: Items available for loan: UVAS Library [Call number: 2877-T] (1).
44.
Detection Of Genetic Variants In Interferon Gamma Gene And Its Association With Resistance Against Mycobacterium Bovis In Buffalo
by Awais Nawaz (2010-VA-219) | Prof. Dr. Asim Aslam | Dr. Muhammad Yasin Tipu | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Bovine Tuberculosis (bovine TB) is a chronic disease of animals and has been known for the significant zoonotic impact. Immune mechanisms necessary for protection against Bovine TB are poorly understood. Interferon-γ cytokine has been reported critically and it is important to study its role in immunity against Bovine TB.
Blood samples were collected from 100 Animals from Peri-urban areas of Lahore, Gujranwala and Okara, Pakistan. Genomic DNA was extracted from the samples. Specific primers were designed to amplify specific portion of IFN-γ gene. Amplified products were sequenced and analyzed by bioinformatics tools. Interferon-γ assay was performed from blood collected in heparin coated vacutainers for the quantification of interferon-γ cytokine in different groups of animals. Blood samples from mycobacterium infected symptomatic and symptomatic animals were processed in Haematology analyzer for complete blood count.
Genetic sequencing of bovine Interferon gamma gene (IFN- γ) help in finding out the Genetic Variations to characterize its role in resistance against Mycobacterium bovis infection. This study help in finding out the confirmed markers for natural resistance against bovine TB that can be used in future selection and breeding programmes. The comparison of hematological values and Interferon gamma level of different groups of animals help us for the detailed diagnosis and prognosis of the disease.
The collected data from hematological analysis of Mycobacterium infected symptomatic animals (Group A), Mycobacterium infected asymptomatic animals (Group B) and non-infected animals/control Group (Group C) was analyzed using the statistical technique of comparing more than two groups i.e. Analysis of variance (ANOVA), One way ANOVA through SPSS 16.0.
CHAPTER 6
SUMMARY
Summary
71
Mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration were found non-significant (p>0.05). White blood cells, Lymphocytes, Platelets, Mean platelet volume and Mean corpuscular volume were found significant (p<0.05). Granulocytes, Red blood cells and Red cell distribution width values were found highly significant.
Interferon gamma assay provided confirmation about the presence of disease in the animals by indicating interferon gamma level to insight the undergoing pathogenesis which was helpful in the detailed diagnosis of the disease. Later on it helped us in the confirmation of false positive results by Tuberculin test.
Final results revealed four intronic variations in different groups of animals. Three of them were found in Group A and B and one was found in Group C (non-infected animals) by Primer 1 (P1). Intronic variations don’t have significant effect but they may have an impact on the regulation of the gene.
We found Transversions (T > A), (A > T), (T > G) were found in mycobacterium infected symptomatic group of animals (Table: 4.7). Transversion (C > G) at and deletion (G >_) was found in this group and exclusive presence of these SNP’s in this group can be considered significant and responsible for the infection. Transversion (A > C) and addition (_ > G) were found in mycobacterium infected asymptomatic group of animals. These two SNP’s are significant as they have been found only in this group. We can infer that the presence of these two SNP's is responsible for the infection along with making them asymptomatic towards the disease.
It was noted that Transition (G > A), (T > C) has been found common in mycobacterium infected symptomatic and asymptomatic group of animals. This common mutation at same position in both groups is quite significant and could be attributed to the occurrence of disease.
Summary Availability: Items available for loan: UVAS Library [Call number: 2881-T] (1).
