PhD Thesis      [Total: 28 ]

Uses of some recent techniques to evaluate the potency of toxins and anti-tetanic serum of c.tetani

Mahmoud Mohammed, 2007

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preparation of antitetanic serumin herses by inoculation of tetanus toxoid alone and tetanus toxoid adjuvanted with aluminium hydroxins gel the tetans toxins and antitetanic serum evaluated by using in familiar techniques as vera cell culture and znzynie lin ked immunosorbent assay (ELISA),agar gel precipitation (AGPT) and fluorescent antibody technique (fat) beside the familiar techniques as flocculation test (f1) and serum neutralization test ( snt). in mice unfamiliar techniques in tetanus field; as ELISA, AGPT, and FAT inaddition to the use of cell cultur could be used accuratelyshowing sensitive, specific and vapid results.


Uses of some recent techniques to evaluate the potency of toxins and antitetanic serum of CI.Tetani

Mahmoud Mohammed, 2007

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Prepaartion of antitetanic serum in horses by inoculation of tetanus toxoid alone and tetanus toxoid adjuvamted with aluminium hydroxide gel. The tetanus toxins and antitetanic serum evaluated by using unfamiliar techniques as Vero cell culture and Enzyme linked Immunosorbent Assay (ELisa), Agar Gel precipitation (AGPT) and Fluorescent antibody technique (FAT), beside the familiar Techniques as fluocculation test (FI)and serum neutralization test (SNT) in mice. Unfamiliar techniques in tetanus field as ELISA, AGPT and FAT in addition to the use of cell culture could be used accurately showing sensitive specific and rapid results.


Further studies on haemorrhagic viral disease in rabbits in Egypt

Owis Salman, 2007

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Sixty one suspected RVHD outbreaks were monitored and diagnosed preliminary in Egypt. The preliminary diagnosis was based on flock history, clinical examination, PM examination, histopathological examination, HAT and virus isolation. RHDV diagnosis was confirmed by carrying out of,SDS-PAGE,Western blotting, RT/PCR assay and IEM. SDS-PAGE and western blotting demonstrated the presence of a major antigen of molecular weight of 50 to 55 KDa approximately. RHDV isolates were tested by RT-PCR using specific primers of three genomic regions encoding the capsid protein (VP60) (regions A and B) and none structural protein (region C), giving positive results, Amplification of VP60 region B from 3 RHDV Egyptian strains, EgyptGiza97, EgyptK2000 and EgyptKalubia 2000, yielded PCR products of 500bp.After sequencing of these products, the sequences were submitted to GenBank with accession numbers EF488823, EF488824 and EF437942 respectively. Amplification of VP60 region A of EgyptKS2000 yielded PCR product of 500bp too. After sequencing of this product, the sequence was submitted to GenBank with accession number EF222287. The obtained nucleotide and amino acid sequence data were compared with that of refernce strains of 17 published on GenBank EHDV sequences, one RCV strain and one EBHSV, RHDV Egyptian strains EgyptGiza97 and EgyptKalubia2000 were genetically and antigenically related and grouped with one genogroup, while RHDV Egyptian strain EgyptKS2000 was genetically differnt and sgregated into different genogroup but antigenically related on the level of VP60 region B. RHDV Egyptian strain EgyptKS2000 was genetically grouped to group 1 , so it is genetically related genetically to the other 2 Egypt strains, while this strain is antigenically different. The pathogenecity of most recently isolated RHDV codeno. R58 (GIZA2006 of HA titer 214 HAU) was carried out using susceptible rabbits. An inactivated vaccine against RVHD was prepared from the most recent local isolate (Giza2006) which identiifed through HAT, IEM and LD50.Comaparative study between the local reecnt inactivated RHDV vaccine and the imported one was carried out. The local vaccine proved to be more immunogenic and protective than the imported one


Further studies on haemorrhagic viral disease in rabbits in Egypt

Owis Salman, 2007

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Sixty one suspected RVHD outbreaks were monitored and diagnosed preliminary in Egypt. The preliminary diagnosis was based on flock history, clinical examination, PM examination, histopathological examination, HAT and virus isolation. RVHD diagnosis was confirmed by carrying out of , SDS-PAGE, Western blotting, RT/PCR assay and IEM. SDS-Page and Western blotting demonstrated the presence of a major antigen of molecular weight of 50 to 55 KDa approximately. RHDV isolates were tested by RT-PCR using specific primers of three genomic regions encoding the capsid protein (VP60) (regions A and B) and none structural protein (region C), giving positive results. Amplification of VP60 region B from 3 RHDV Egyptian strains, EgyptGiza97, Egypt KS 2000 and Egypt Kalubia 2000, yielded PCR products of 500 bp. After sequencing of these products, the sequences were submitted to GenBank with accession number EF488823, EF 488824 and EF 437942 respectively. Amplification of VP60 region A of EgyptKS 2000 yielded PCR product of 500 bp too. After sequencing of this product, the sequence was submitted to Genbank with accession number EF22287. The obtained nucleotide and amino acid sequence data were compared with that of reference strains of 17 published on Genbank RHDV sequences, one RCV strain and one EBHSV. RHDV Egyptian strains EgyptGiza 97 and EgyptKalubia2000 were genetically and antigenically related and grouped within one genogroup, while RHDV Egyptian strain EgyptKS2000 was genetically different and segregated into different genogroup but antigenically related on the level on the level of VP60 region B.RHDV Egyptian strain EgyptKS2000 was genetically grouped to group 1, so it is related genetically to the other 2 Egyptian strains, while this strain is antigenically different. The pathogenicity of most recently isolated RHDV code no. R58 (Giza2006) of HA titer 214 HAU) was carried out using susceptible rabbits. An inactivated vaccine against RVHD was prepared from the most recently local isolate (Giza 2006) which identified through HAT, IEM and LD50. Comparative study between the local recent inactivated RHDV vaccine and the imported one was carried out. The local vaccine proved to be more immunogenic and protective than the imported one.


Further studies on infectious larynogtracheitis in chicken flocks”

Namaa Abd El-hafiz, 2006

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Infectious laryngotracheitis virus was detected from outbreaks in kalubia , Sharkia and gharbia governorates. The isolates were subjected to physicochemical, biological characterization and pathological investigations. Pathogenicity test showed that isolate S3 was the most pathogenic strain.The S3 isolate was used for preparation of inactivated ILTV vaccine by using Binary Ethelinimine, with Aluminium hydroxide gel and montanide ISA -70M as adguvant. The cell mediated and humoral immune response, and potency test of the vaccinated birds were indicated that ILT prepared inactivated vaccine with gel and oil adjuvants are potent for birds’ protection as will as the live vaccines, and they had no adverse post- vaccinal reaction.


Trials for preparation and evaluation of different vaccines against enteropathogenic escherochia coli in rabbits

Eman Ahmed Zaki, 2006

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The aim of this thesis was to study the incidence and mortality rates in diarrhoeic rabbits suffering from E. coli infection. correlation between biotyping and serotyping of E. coli isolates revealed the following: O8/1-; O20/1+; O26/2+; O15/3-; O153/5+ and O12/8+ virulence attribute confirrned that these strains were belonging to enteropathognic E. coli. The comparative protective capacity of the ultrasonicated, irradiated, heat inactivated, forinalinized inactivated as attenuated –irradiated vaccines was elucidated using challenge test, local , IgA antibody response as well as immunohistopathological response. Results indicated that the ultrasonicated vaccine was the best vaccine for protective against E. coli infection in rabbits.


Trials for vaccine preparation against tuberculosis

Mohamed Osman, 2006

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ESAT-6 is a secreted protein present in the short term culture filtrate of M. tuberculosis after growth on a synthetic Sauton medium. It induces high potent T- cell response and production of IFN-y which plays a critical role in protective cell mediated immunity against tuberculosis. Its corresponding gene is located in RD1 deleted in BCG vaccine. The current study aimed for the cloning and expression of ESAT-6 protein antigen from M.bovis and its use alone or plus adjuvants as a vaccine against bovine tuberculosis genomic DNA was exctracted using triazol, purified, concentrated and esat -6 was amplified by PCR and assessed by agarose gel electrophoesis esat-6 was ligated to an expression vector, PQE. After transformation by electroporation of the expression host E.coli with the recombinant PQE plasmid, the expression was induced using 10 mM of IPTG, two bands were seen in the SDS-page analysis, the 18 kDa which represented the dimmer forms of their ESAT-6 antigen the histagged r-ESAT-6 antigen was then purified by metal affinity chromatography using Ni-NTA agarose. Four groups of albino guinea pigs each group 10 animals were vaccinated, the first using r-ESAT-6 +dimethyl dioctadecyl ammonium bromide (DDA). The second with r-ESAT-6. the third with BCG vaccine, the fourth with DDA, and there was another group non vaccinated (control). A booster dose of the vaccine was given after 2 weeks and route of vaccination was s/c Two weeks later challenge was done by infecting all groups with M.bovis I/N. dose of vaccination was 0.2ml and dose of challenge was 1 ml. Evaluation of the vaccine was done by : Determination of immunogenicity of prepared antigen by ELISA techniques that was applied after the booster dose of vaccine and 2 weeks post challenge and it was found that it gave high immunogenicity with serum of guinea pig groups vaccinated with r-ESAT-6 alone or plus DDA. Nearly the same results obtained after challenge. Determination of cell proliferation using Brdu ELISA kits and it was found that cell proliferation was observed in Guinea pigs groups vaccinated with BCG, r ESAT – 6 and ESAT-6 +DDA. Other investigations were applied to detect the potency of the vaccine : 1- Survival rate was higher in groups vaccinated with rESAT 6 +DDA (70%), rESAT-6 (70%) and BCG (80%) while it was in DDA (30%) and zero percent in negative control group. 2- The viable bacterial count was determined in tissues of lungs and spleens and it was found higher in negative control and DDA then decrease respectively in rESAT-6 then rESAT-6+DDA and BCG vaccinated groups. The results were found in spleen parallel to that of lung except that it was higher in DDA than negative control group. 3- RSLW when exceeds indicate infection with M.bovis 4- Lung density determination and the degree of sinking and floating. When float indicate low rate of infection and when sink indicate high rate of infection Finally, the histopathological findings approved the previous results. Our study suggests that ESAT-6 is an antigen with major potential for vaccination against and specific diagnosis of bovine tuberculosis


Attempts to produce a vaccine against Avian spirochaetosis

lilian Saad, 2005

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In this work a lot of efforts had been done for propagation of Brosella anserina microorganism was applied in this present work either by Cuffirating themm in the stablized BSKII culture medium which considered specific medium for Borrelia anserina or by propagating them in the embryonateed chicken eggs via chori-allantoic sac . Five passages of Borrelia anserina culture were obtained using microaerophilic method of inoculation , while seven passages were recorded by anaerobic route of incubation. Two types of vaccine adjuvants have been used in the present work which were hydroxid gel (alhydragel) adjuvant and oil (emulsion) adjuvant, good and satisfied results were recorded. It was found through this present study that the addition of adjuvants to the formalized killed vaccines improve the quality of the vaccine and lead to increase the stability, keeping quality and immunity. Culture vaccines against Avian spirochaetosis have prepared for the first time in this study. According to this work, it was found that the culture vaccines have higher immunizing power and longer keeping quality than egg vaccine Also through this study it was decided that oil emulsion adjuvant provided higher immunity response and more stable thatn hydroxid gel adjuvant. The efficacy and degree of immunity of the sear of vaccintaed birds wer measured by both ELISA and serum plate agglutination test. From this study it was found that ELSIA test is very accurate and sensitive. The prepared vaccines were administred as two divided doses with 3 weeks interval.


دراسات فيروسية على مجموعة جدري الكابري

Abd Elrazek Omar, 2003

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Immunological studies on the locally prepared Bivalent vaccine of ‎spirochaeta and newcaste disease

2003

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Spirocheata and NewCastle disease are the most important respiratory and ‎disease affecting poultry and cause a great economic losses due to highly ‎rate of morbidity and mortality as well as drop in egg production so the aim ‎of this study is to produce a locally combined vaccine / ND v and ‎spirochaeta against both disease for a aving tie , effort and money the ‎evaluation of the all vaccines (combined / NDV vaccine / spirochaeta) for ‎the priority and safety , and all vaccine were free from any bacterial , virus , ‎fungal contaminants the serologicaly examination to sera sample collected ‎from chickens vaccinated by the combinedvaccine (NDV and spirochaeta) , ‎lyophilized spirochaeta vaccine and formalized inactivated N.D.V was done ‎by selected serological test H1 test , SNT test slid agglutination test and ‎ELISA test. Also came be evaluation the cellular immune response by ‎lymyhocte trans formation test by using the mitogen. One for B. Cell and the ‎other for T. Cell. All this test used for the evaluation of the immune ‎response and immunity which a quired to bird from vaccinated with all ‎vaccinated mention befor. All result of locally prepared combined vaccine ‎give highly and stable immune response than the NDV vaccine and ‎spirochaeta vaccine especially in the last part of the experiment. Due to the ‎wide spread of spirochaeta among fowls in Egypt . locally prepared ‎combined vaccine can be used safely in vaccination against both disease ‎‎(spirochaeta and NDV). Early vaccination against both disease are urgent ‎demand in poultry industry as the present study proved that the combined ‎vaccine can be inoculated safely at end of the first week in chicks ‎


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