) قالوا سبحانك لا علم لنا الا ما علمتنا انك انت العليم الحكيم ( البقرة 32

2. )قالوا سبحانك لا علم لنا الا ما علمتنا انك انت العليم الحكيم( البقرة 32

3. A Trial for Prevention of Campylobacter jejuni Infection in Broiler Chickens using Autogenus Bacterin M. H. Awaad1, Nagwa, S. R.2 and Wafaa A. Abd El-Ghany1 1Faculty of Veterinary Medicine, Cairo University 2National Research Centre, Egypt

4. Campylobacter jejuni (C.jejuni) infection in chickens has been implicated as a disease characterized by chronic course, high morbidity, low mortality and reduction in egg production (Peckham,1984). C.jejuni is the primary cause of human gatro-intestinal infection among campylobacter species. Consumption of chickens and retail poultry products is incriminated as the main vehicle for transmission of infection to human (Nachamkin et al.,1993).

5. So, it is important to address intervention strategies by which these bacteria can be reduced or removed from the food animals. Such strategies could include vaccines. In chickens, different types of living and killed vaccines against C.jejuni have been used with variable results (Stern et al.,1990;Cowthrow et al.,1994;Rice et al.,1997;Lee et al.,1999 and Ziprin et al.,2002).

6. The aim of the study Preparation of two types of C.jejunibacterins from the local field strains and evaluation of their protective potential in broiler chickens through immuno-Assay [mean Enzyme Linked Immuno-sorbent Assay (ELISA) titres] and bio-Assay (clinical signs, mean lesion score, rate of shedding, reisolation rate and histopathological examination)

7. Materials & Methods Three local strains of C.jejuni representing biotype I and II were used in bacterins preparation and as a challenge bacteria. Preparation of whole cell bivalent C.jejunibacterinsaluminium hydroxide (AHAB) and incomplete Freund’s oil adjuvant bivalent bacterins(IFAB) )109CFU/ml C.jejuni) as described byWilliams et al., 1976 and Bryner et al., 1978 and 1988. Quality control tests of the prepared bacterins (purity, sterility and safety) tests (British veterinary codes, 1970)

8. Experimental design

10. Results of quality control tests of bivalent C.jejuni bacterins 1. Purity test Microscopical and biochemical identification of the grown seed culture onto Skirrow’s media revealed presence of pure culture of C.jejuni cells. 2. Completion of C.jejuni inactivation Inactivated C.jejuni cells showed no growth on Skirrow’s agar plates after formalin inactivation.

11. 3. Sterility test The formalin killed C.jejuni cells gave no bacterial growth after cultivation onto PPLO agar and broth and also no fungal growth was obtained after cultivation onto sabouraud dextrose agar plates. 4. Safety test The prepared bivalent bacterins were found to be safe for day-old-chicks, producing neither clinical signs nor local reactions and deaths during seven successive days observation period.

12. Comparison between the results of ELISA in immunized chicken broilers with (AHAB) and (IFAB) at 1 & 3 wks of age

13. Results of Bio-Assay Clinical signs Signs of depression, sleepy appearance and mucoid greenish diarrhea were seen only in control C.jejuni challenged group. Mortalities No deaths were recorded in all C.jejuni challenged groups.

14. Shedding of C.jejuni in immunized and unimmunized broiler chickens during 21 days observation period

15. Reisolation of C.jejuni from immunized and unimmunized chicken broilers at the end of 21 days observation period

16. Gross lesion scoring of C.jejuniin immunized and un-immunized sacrificed chicken broilers at the end of 21 days observation period.

17. The main histopapatological lesions in liver and degree of severity in chicken broilers immunized with two types of C.jejunibacterins ++++=Severe +++=Moderate ++ & +=Mild

18. The main histopapatological lesions in intestine and degree of severity in chicken broilers immunized with both bacterins ++++=Severe +++=Moderate ++ & +=Mild

19. 1 2 4 3 Fig. (1): Liver of broilers immunized with (IFAB)and infected with C.jejunishowing few numbers of inflammatory cells in blood vessels. (H & E X33). Fig. (2): Liver of broilers immunized with (AHAB)and infected with C.jejunishowing focal area of necrosis (arrow). (H & E X33). Fig. (3): Liver of broilers immunized with (IFAB) and infected with C.jejunishowing mild vacculation of hepatocytes. (H & E X66). Fig. (4): Liver of control(unimmunized and infected) broilers showing severe hyperplasia of the epithelial lining the bile duct. (H & E X66).

20. 5 6 7 Fig. (5): Intestine of control (unimmunized and infected) broilers showing severe hyperplasia of enterocytes (arrow). Notice severe inflammatory cells aggregation in lamina propria. (H & E X66). Fig. (6): Intestine of broilers immunized with (IFAB) and infected with C.jejuni showing mild hyperplasia of enterocytes, goblet cells transformation (arrow) and mild inflammatory cells aggregation. (H & E X33). Fig. (7): Intestine of broilers immunized with (AHAB) and infected with C.jejuni showing hyperplasia of crypt of Luberkuhn with moderate numbers of mitotic figures (arrow).(H & E X66).

21. CONCLUSION • Application of C.jejuniwater-type or oil-type bacterin was effective considering stimulation of immune response, reduction of signs, lesions and the organism shedding. • Oil type bacterin was more effective than water type one.

22. Thank you Wafaa Abd El-Ghany