Identification And Molecular Characterization Of Shiga Toxin Producing
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Publisher: 2011 Dissertation note: Escherichia coli is normal inhabitant of all the animals and human beings. Its non-sorbitol fermenting biotype (SNF) was detectable in buffalo (90 percent), cattle (80 percent) or rarely in sheep (20 percent) and goat (30 percent). However, SNF E. coli were un-detectable in droppings of rural chickens and feces of donkeys. The SNF E. coli was detected in 100, 92, 71 and 80 percent of the market milk samples and 100, 83, 83 and 53 percent beef samples from Multan, Sandha, Wagha and Sheikhupura Roads, of Lahore city, respectively. However, SNF E. coli was not detected from freshly aseptically collected milk and beef samples but was detectable in over all 96 percent of market raw milk and 82 percent market beef samples. White colored colonies of each positive sample on Sorbitol MacConkey's Agar (SMA) contained gram negative, rods with round ends non spore former. Each of the isolates was sorbitol non fermenter, lactose fermenter, indole positive, Methyle Red positive, Voges Prauskaur negative and citrate negative. However, each of such isolates showed green metallic sheen on Eosin Methylene Blue (EMB) agar.
Each of the isolate was further characterized using polymerase chain reaction (PCR). Bacterial DNA was extracted easily by boiling method when the isolates were grown on Sorbitol MacConkey's agar or EMB agar at 370C for 12-24 hours. The DNA was recovered when the culture was boiled for 5-10 minutes. The DNA of each sample remained stable on storage at 40C for 48 hours or at -200C for 7 days. The isolated DNA (100 samples) when amplified using universal, Stx1, Stx2 and O157 specific primers showed that 82 percent samples were positive for universal primers, 50 percent for O157, 60 % for Stx1 and 51 percent for Stx2. The filtrate of each isolate when diluted as 1:10 dilution and sterilized by filtration induced cyto-pathogenic effect (CPE) on Vero cell line.
It is concluded that SNF E. coli O157 normally exists in intestinal tract of buffalo, cattle, sheep and goat. Counts of SNF E. coli O157 were higher in milk samples as compared to beef samples. More than 80 percent samples of milk or beef were contaminated with SNF E. coli O157. Feces of the animals are presumably main source of SNF E. coli contamination of raw milk and beef. PCR is a quick, reliable, and sensitive technique for confirmation of SNF E. coli O157 in the samples.
Availability: Items available for loan: UVAS Library [ Call number: 1221,T] (1).
Comparative Efficacy Of Rocombinant Avian Influenza H9N2 Vectored Vaccine In Poultry
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Publisher: 2011 Dissertation note: A correlative study testing the impact of different urban congestions in Lahore upon the bird abundance and diversity was conducted in the monsoon and winter season. A platform of 6 feet diameter and 4 feet height, with a variety of seeds, was offered to attract the grainivorous birds. The number and variety of birds visiting the feeding station was noted, from Dawn to dusk after a period of 6 day installation, which depicted the bird's urbanization in the area. A significant increase in the abundance of birds was observed in the winter season. Areas with large sized pockets of vegetation supported the largest diversity of birds. Grain preference of the birds recorded in the different areas studied showed a relationship between the age long practices of feeding birds with the habituation of the birds.
In order of Preference Birds of most areas preferred Pearl Millet (Bajra) seeds, and Rice Seeds, over others. The next choice of food was shown to be Italian Millet and Red Italian Millet, when the amount of these decreased birds started feeding on, Sunflower, then Safflower, and Wheat. Mustard, corn and Chick peas remained feed of least preference. A weak correlation was found between urban population congestion and the number of pigeons, doves, babblers, house sparrows and silver bills. These birds are considered to be urbanized.
As shown in the Tables the results suggest that more grainivorous species and invasive species such as House Sparrows are present in the central parts of the city. The outskirts of Lahore, the sub urban areas enjoy a large diversity of avifauna. This finding is supported by Mathew and Naik (1993). Areas where a considerable amount of vegetation is present, such as parks, gardens or big lawns with a lot of vegetation provide a good habitat can be considered as "Pockets of Wilderness" These areas, provide food and habitat for a variety of bird species. Although an exponential increase in urban congestion of cities and a need for further development does not allow room for large areas supporting natural vegetation. However, a plan has to be developed which would ensure a set amount of area for vegetation, to support a healthy diversity of bird species.
Availability: Items available for loan: UVAS Library [ Call number: 1265,T] (1).
Dynamics Of Recombinant Avian Influenza Virus H9-Ha Gene Herpesvirus Of Turkey Vector Vaccine In Chicken
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Publisher: 2010 Dissertation note: Avian influenza (AI) is known to exist for centuries. It's primarily a disease of birds. For prevention and control of H9N2 avian influenza disease, HA gene from Pakistani H9N2 field virus isolate (PK-UDL/01/08 H9N2) cloned into a herpesvirus of turkey (rHVT/AI-H9 having full HA) at Institute for Animal Health, Compton, UK, expressing HA proteins, was given subcutaneously (in the neck). Commercial avian influenza vaccine was given to chicken through intramuscular route.
On termination of the experiment, the chicks from all groups were sacrificed and their visceral organs were collected on chick-wise basis. Besides this, blood was collected for complete blood counts (CBC) and blood chemistry. Results of groups were then compared for any significant difference. The data analyses showed that in complete blood counts, there was no significant difference (p<0.05) of total leukocyte counts of different groups of chickens but heterophil percentages showed variation among different groups.
Analyses of Serum chemistry results showed that glucose and protein concentration in serum varies significantly (p>0.05) among the different groups of birds. However, there was no significant difference of cholesterol levels among the groups of chickens.
For determining the persistence of rHVT/AI-H9 having full HA virus in different visceral organ and in blood samples of Group A, results of the PCR showed the persistence of herpes turkey virus in leukocytes, spleen, liver, lung, kidney and heart. This project helped in evaluating dynamics of recombinant HVT containing avian influenza HA gene from avian influenza H9N2 Pakistani isolate.
In future the study may be continued for further biological characterization by isolating the HVT from different visceral organ in different interval of time and fractionating the serum protein and analyzing the gamma globulin fraction in serum to measuring the humoral immune response against the recombinant vaccine.
Availability: Items available for loan: UVAS Library [ Call number: 1276,T] (1).
Effectof Mentofin (Herbal Product)On Antibody Response Of Broilers To Newcastle Disease Vaccine
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Publisher: 2011 Dissertation note: Immunostimulants such as Mentofin® are commonly used to enhance the immune response of birds to vaccines. In the present study immunomodulatory effect of Mentofin® on antibody response of broilers to Newcastle Disease virus vaccine was evaluated. For this purpose one hundred one day old broilers were divided in to four groups (A, B, C and D) each containing 25 birds. Each bird of group A and B was vaccinated against ND and each bird of group A and C was treated with Mentofin. Anti-ND-HI antibody titer of each bird of each group was monitored on 14, 21, 28 and 35 days of age. Mentofin® treated broilers showed higher consistent antibody (anti-NDV HI antibody titer) response as compared to untreated broilers. These birds when given challenge infection of velogenic ND virus on 35 days of age showed same protection as that of untreated vaccinated birds. However non vaccinated broilers treated with Mentofin showed higher protection as compared to that of in non treated unvaccinated birds. Weight gain in Mentofin® treated broilers was same as that of non-treated birds. Similarly, there was no effect of the Mentofin on FCR. Droppings from Mentofin treated birds showed no urease producing bacteria while 100% droppings of the herbal untreated birds showed urease producing bacteria. Mentofin at 1% concentration in nutrient broth inactivated the proteus species while its 0.0001 % concentration inactivated the bacteria in urea broth. In in vitro studies, 0.5 % concentration of Mentofin inactivated the lentogenic strain of ND Virus within 15 minutes at interaction temperature of 37 0C.
The results can be used to formulate the vaccination policy along with herbal based immunostimulatant such as Mentofin®.
Availability: Items available for loan: UVAS Library [ Call number: 1294,T] (1).