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Study Of Pathogenesis Of Mycoplasma Gallisepticum In White Leg Horn Layer

by Mubasher Rauf | Prof.Dr.Zafar Iqbal Ch | Dr Aftab | Dr.M.Younus Rana.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: In first part of present study 380 samples were collected from clinically suspected cases of layers suffering from respiratory diseases in and around Lahore. Samples were subjected for mycoplasma isolation by using Frey's medium. Plates with positive growth revealed characteristic colonies on 8th day post inoculation that reached maximum in size and growth at 15th day post inoculation. Out of 380 samples, 104 (27.36%) samples were positive on culture. Isolates were identified through growth inhibition test (GIT) by using hyper immune sera raised in rabbits. Isolates were further confirmed by PCR. Similarly, tracheal swabs and tissue samples of lungs and trachea collected under refrigeration were also subjected for DNA analysis. Out of 380 samples 264 (69.5%) were positive on PCR analysis. By comparing two diagnostic techniques it was found that PCR was more sensitive and reliable technique for screening of Mycoplasma gallisepticum. In second part of study experiment isolates were analyzed for protein profile of Mycoplasma by standardization of two techniques Sodium dodecyl sulphate polyacrylamide gel (SDS-PAGE) and western blotting. These techniques help to find out any antigenic variation in prevailing strain of mycoplasma. During our study five bands of protein were detected with molecular size of 32.35 kDa, 43.65 kDa, 52.48 kDa, 64.56 kDa and 70.8 kDa. These proteins were extracted from whole cell of Mycoplasma gallisepticum isolates. On comparing the molecular sizes it was found that isolated species showed low antigenic variation, analyzed by SDS-PAGE. Western blot was used to determine the specific protein of Mycoplasma gallisepticum with the use of specific polyclonal antibody raised in rabbit. The positive reaction site was shown on nitrocellulose membrane confirming target species of CRD. During third part of present study, it was concluded that aerosol route of infection causes early disease, followed by intra tracheal and per-oral route respectively. The severity of infection was found more in aerosol and intra tracheal routes of inoculation than per-oral route which was found to be very mild. The general gross lesions observed in the above two groups were hemorrhages in trachea with mucous plug. There was air sacculitis, hemorrhages in the lungs, salpingitis and putrefied eggs in the ovary. On histopathological examination lesions were found in trachea, lungs and oviduct. Re-isolation was carried out to confirm antigen in experimentally inoculated birds. Paraffin embedded sections of trachea, lungs and oviduct were processed for immunohistochemical examination in order to confirm the antigen of Mycoplasma gallisepticum within tissue. A positive immunochemical reaction was found in lungs and oviduct. Which represents that antigen was same as inoculated during study of pathogenesis. Availability: Items available for loan: UVAS Library [Call number: 1258,T] (1).

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Toxico-Pathological And Hematological Study In Japanese ( Coturnix Coturnix Japonica ) Exposed To Ochratoxin A And Aflatoxin B

by Muhammad shahzad | Dr. Muti-ur-rehman khan | Dr. kamran | Dr. M. younus rana.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2011Dissertation note: Mycotoxins are secondary metabolite toxins produced by fungi in or on grains, cereals and nuts used as feed in the poultry industry. Mycotoxicity in birds has been well documented and its severity increases in combination with other toxins. The study determined synergistic pathological responses in quail chicks when fed different level of Aflatoxin B1 (AFB1) and Ochratoxin A (OTA). A total of 245 quails chicks were divided into seven groups (G1-G7) having 35 quail chicks in each. OTA mixed feed was fed to quail chicks at a dose rate of 1 and 1.5 ppm in G1 and G2 respectively. G3 were fed 2 ppm OTA + 1 ppm AFB1. AFB1 was offered in the feed at a level of 1 and 1.5 ppm in G4 and G5 respectively. G6 birds were fed 2 ppm AFB1 + 1 ppm OTA while G7 acted as a control. All the birds offered toxin free basal diet for first 7 days. Day 7 was considered zero day of experiment. At this day chicks shifted to different groups of 35 each (G1-G7). Group G7 was control group and offered toxin free diet. Birds were monitored twice daily for clinical signs. Randomly selected six birds from each group were slaughtered at day 14, 21, and 28. Blood samples with and without anticoagulant were collected for hematological and biochemical studies respectively. Morbid tissue of liver, kidney, and intestine were collected for histopathological studies. The OTA groups developed anemia manifested by a significant decrease in the red blood cell count, packed cell volume percentage and hemoglobin concentration, while increase erythrocyte sedimentation rate at the end of the experiment all groups showed significant reduction in red blood cell count. This reduction was found to increase with time proportionally to the level of OTA and AFB1 alone or in combination exposure. Clinical signs in chicks administered AFB1 and OTA included depression, decreased feed intake and decreased body weight. Severity in clinical signs was dose related. Pathological lesions in liver of these chicks were hemorrhages, fatty change, centrilobular necrosis and periportal fibrosis. Microscopically, liver showed vacuolation, fatty change, congestion and individual cell necrosis. Kidney of these chicks included pyknotic changes in the epithelium of proximal and distal convoluted tubules. Severe necrotic changes in the collecting ducts and accumulation of pink homogenous material in the lumen of tubules. Intestine showed hemorrhages, edema, degeneration and infiltration of mononuclear cells were observed. OTA damaged intestinal mucosa more severely than AFB1. Serum biochemical study indicated a significant decrease in total serum proteins and increase in urea and creatinine. It is concluded that AFB1 and OTA are capable of inducing hematological and histopathological alterations in quail chicks at higher dietary concentrations, either individually or in combination. Availability: Items available for loan: UVAS Library [Call number: 1301,T] (1).

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